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Sample records for matrix-assisted pulsed laser

  1. Lipase biofilm deposited by Matrix Assisted Pulsed Laser Evaporation technique

    Aronne, Antonio; Bloisi, Francesco; Calabria, Raffaela; Califano, Valeria; Depero, Laura E.; Fanelli, Esther; Federici, Stefania; Massoli, Patrizio; Vicari, Luciano R.M.

    2015-01-01

    Highlights: • A lipase film was deposited with Matrix Assisted Pulsed Laser Evaporation technique. • FTIR spectra show that laser irradiation do not damage lipase molecule. • Laser fluence controls the characteristics of complex structure generated by MAPLE. - Abstract: Lipase is an enzyme that finds application in biodiesel production and for detection of esters and triglycerides in biosensors. Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique derived from Pulsed Laser Deposition (PLD) for deposition of undamaged biomolecules or polymers, is characterized by the use of a frozen target obtained from a solution/suspension of the guest material (to be deposited) in a volatile matrix (solvent). The presence of the solvent avoids or at least reduces the potential damage of guest molecules by laser radiation but only the guest material reaches the substrate in an essentially solvent-free deposition. MAPLE can be used for enzymes immobilization, essential for industrial application, allowing the development of continuous processes, an easier separation of products, the reuse of the catalyst and, in some cases, enhancing enzyme properties (pH, temperature stability, etc.) and catalytic activity in non-aqueous media. Here we show that MAPLE technique can be used to deposit undamaged lipase and that the complex structure (due to droplets generated during extraction from target) of the deposited material can be controlled by changing the laser beam fluence

  2. Lipase biofilm deposited by Matrix Assisted Pulsed Laser Evaporation technique

    Aronne, Antonio [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Bloisi, Francesco, E-mail: bloisi@na.infn.it [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy); Calabria, Raffaela; Califano, Valeria [Istituto Motori – CNR, Naples (Italy); Depero, Laura E. [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Fanelli, Esther [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Federici, Stefania [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Massoli, Patrizio [Istituto Motori – CNR, Naples (Italy); Vicari, Luciano R.M. [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy)

    2015-05-01

    Highlights: • A lipase film was deposited with Matrix Assisted Pulsed Laser Evaporation technique. • FTIR spectra show that laser irradiation do not damage lipase molecule. • Laser fluence controls the characteristics of complex structure generated by MAPLE. - Abstract: Lipase is an enzyme that finds application in biodiesel production and for detection of esters and triglycerides in biosensors. Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique derived from Pulsed Laser Deposition (PLD) for deposition of undamaged biomolecules or polymers, is characterized by the use of a frozen target obtained from a solution/suspension of the guest material (to be deposited) in a volatile matrix (solvent). The presence of the solvent avoids or at least reduces the potential damage of guest molecules by laser radiation but only the guest material reaches the substrate in an essentially solvent-free deposition. MAPLE can be used for enzymes immobilization, essential for industrial application, allowing the development of continuous processes, an easier separation of products, the reuse of the catalyst and, in some cases, enhancing enzyme properties (pH, temperature stability, etc.) and catalytic activity in non-aqueous media. Here we show that MAPLE technique can be used to deposit undamaged lipase and that the complex structure (due to droplets generated during extraction from target) of the deposited material can be controlled by changing the laser beam fluence.

  3. Comparative study on Pulsed Laser Deposition and Matrix Assisted Pulsed Laser Evaporation of urease thin films

    Smausz, Tomi; Megyeri, Gabor; Kekesi, Renata; Vass, Csaba; Gyoergy, Eniko; Sima, Felix; Mihailescu, Ion N.; Hopp, Bela

    2009-01-01

    Urease thin films were produced by Matrix Assisted Pulsed Laser Evaporation (MAPLE) and Pulsed Laser Deposition from two types of targets: frozen water solutions of urease with different concentrations (1-10% m/v) and pure urease pellets. The fluence of the ablating KrF excimer laser was varied between 300 and 2200 mJ/cm 2 . Fourier transform infrared spectra of the deposited films showed no difference as compared to the original urease. Morphologic studies proved that the films consist of a smooth 'base' layer with embedded micrometer-sized droplets. Absorption-coefficient measurements contradicted the traditional 'absorptive matrix' model for MAPLE deposition. The laser energy was absorbed by urease clusters leading to a local heating-up and evaporation of the frozen matrix from the uppermost layer accompanied by the release of dissolved urease molecules. Significant enzymatic activity of urease was preserved only during matrix assisted transfer.

  4. Production of active lysozyme films by matrix assisted pulsed laser evaporation at 355 nm

    Purice, Andreea; Schou, Jørgen; Kingshott, P.

    2007-01-01

    Thin lysozyme films have been produced in a dry environment by MAPLE (matrix assisted pulsed laser evaporation) from a water ice matrix irradiated by laser light at 355 nm above the absorption threshold of the protein. A significant part of the lysozyme molecules are transferred to the film without...

  5. Processing of C60 thin films by Matrix-Assisted Pulsed Laser Evaporation (MAPLE)

    Canulescu, Stela; Schou, Jørgen; Fæster, Søren

    2011-01-01

    Thin films of fullerenes (C60) were deposited onto silicon using matrix-assisted pulsed laser evaporation (MAPLE). The deposition was carried out from a frozen homogeneous dilute solution of C60 in anisole (0.67 wt%), and over a broad range of laser fluences, from 0.15 J/cm2 up to 3.9 J/cm2. MAPLE...

  6. Hydroxyapatite thin films grown by pulsed laser deposition and matrix assisted pulsed laser evaporation: Comparative study

    Popescu-Pelin, G.; Sima, F.; Sima, L. E.; Mihailescu, C. N.; Luculescu, C.; Iordache, I.; Socol, M.; Socol, G.; Mihailescu, I. N.

    2017-10-01

    Pulsed Laser Deposition (PLD) and Matrix Assisted Pulsed Laser Evaporation (MAPLE) techniques were applied for growing hydroxyapatite (HA) thin films on titanium substrates. All experiments were conducted in a reaction chamber using a KrF* excimer laser source (λ = 248 nm, τFWHM ≈ 25 ns). Half of the samples were post-deposition thermally treated at 500 °C in a flux of water vapours in order to restore crystallinity and improve adherence. Coating surface morphologies and topographies specific to the deposition method were evidenced by scanning electron, atomic force microscopy investigations and profilometry. They were shown to depend on deposition technique and also on the post-deposition treatment. Crystalline structure of the coatings evaluated by X-ray diffraction was improved after thermal treatment. Biocompatibility of coatings, cellular adhesion, proliferation and differentiation tests were conducted using human mesenchymal stem cells (MSCs). Results showed that annealed MAPLE deposited HA coatings were supporting MSCs proliferation, while annealed PLD obtained films were stimulating osteogenic differentiation.

  7. The minimum amount of "matrix " needed for matrix-assisted pulsed laser deposition of biomolecules

    Tabetah, Marshall; Matei, Andreea; Constantinescu, Catalin

    2014-01-01

    The ability of matrix-assisted pulsed laser evaporation (MAPLE) technique to transfer and deposit high-quality thin organic, bioorganic, and composite films with minimum chemical modification of the target material has been utilized in numerous applications. One of the outstanding problems in MAPLE...

  8. Characterization of lysozyme films produced by matrix assisted pulsed laser evaporation (MAPLE)

    Purice, Andreea; Schou, Jørgen; Kingshott, Peter

    2007-01-01

    Thin lysozyme films of thickness up to more than 100 nm have been produced in a dry environment by MAPLE (matrix assisted pulsed laser evaporation) from a water ice matrix. Analysis of the films demonstrates that a significant part of the lysozyme molecules is transferred to the substrate without...

  9. Surface morphology of thin lysozyme films produced by matrix-assisted pulsed laser evaporation (MAPLE)

    Purice, Andreea; Schou, Jørgen; Pryds, Nini

    2007-01-01

    Thin films of the protein, lysozyme, have been deposited by the matrix-assisted pulsed laser evaporation (MAPLE) technique. Frozen targets of 0.3-1.0 wt.% lysozyme dissolved in ultrapure water were irradiated by laser light at 355 mn with a fluence of 2 J/cm(2). The surface quality of the thin....... The concentration of lysozyme in the ice matrix apparently does not play any significant role for the morphology of the film. The morphology obtained with MAPLE has been compared with results for direct laser irradiation of a pressed lysozyme sample (i.e. pulsed laser deposition (PLD)). (C) 2007 Elsevier B.V. All...

  10. Growth of thin films of low molecular weight proteins by matrix assisted pulsed laser evaporation (MAPLE)

    Matei, Andreea; Schou, Jørgen; Constantinescu, C.

    2011-01-01

    Thin films of lysozyme and myoglobin grown by matrix assisted pulsed laser evaporation (MAPLE) from a water ice matrix have been investigated. The deposition rate of these two low molecular weight proteins (lysozyme: 14307 amu and myoglobin: 17083 amu) exhibits a maximum of about 1–2 ng/cm2 per....... The results for lysozyme demonstrate that the fragmentation rate of the proteins during the MAPLE process is not influenced by the pH of the water solution prior to freezing....

  11. UV and RIR matrix assisted pulsed laser deposition of organic MEH-PPV films

    Toftmann, B.; Papantonakis, M.R.; Auyeung, R.C.Y.; Kim, W.; O'Malley, S.M.; Bubb, D.M.; Horwitz, J.S.; Schou, J.; Johansen, P.M.; Haglund, R.F.

    2004-01-01

    A comparative study of thin film production based on gentle laser-ablation techniques has been carried out with the luminescent polymer poly[2-methoxy-5-(2'-ethylhexyloxy)-1,4-phenylene vinylene]. Using a free-electron laser films were made by resonant infrared pulsed laser deposition (RIR-PLD). For the first time resonant infrared matrix assisted pulsed laser evaporation (RIR-MAPLE) was successfully demonstrated on a luminescent polymer system. In addition to this, an excimer laser has been used for UV-MAPLE depositions at 193 and 248-nm irradiation. Films deposited onto NaCl and quartz substrates were analyzed by Fourier transform infrared spectroscopy, UV-visible absorbance and photoluminescence. Photoluminescent material was deposited by RIR-MAPLE and 248-nm MAPLE, while the RIR-PLD and 193-nm-MAPLE depositions displayed the smoothest surfaces but did not show photoluminescence

  12. UV and RIR matrix assisted pulsed laser deposition of organic MEH-PPV films

    Christensen, Bo Toftmann; Papantonalis, M.R.; Auyeung, R.C.Y.

    2004-01-01

    -PLD). For the first time resonant infrared matrix assisted pulsed laser evaporation (RIR-MAPLE) was successfully demonstrated on a luminescent polymer system. In addition to this, an excimer laser has been used for UV-MAPLE depositions at 193 and 248-nm irradiation. Films deposited onto NaCl and quartz substrates......A comparative study of thin film production based on gentle laser-ablation techniques has been carried out with the luminescent polymer poly [2-methoxy-5-(2'-ethylhexyloxy)-1,4-phenylene vinylene]. Using a free-electron laser films were made by resonant infrared pulsed laser deposition (RIR...... were analyzed by Fourier transform infrared spectroscopy, UV-visible absorbance and photoluminescence. Photoluminescent material was deposited by RIR-MAPLE and 248-nm MAPLE, while the RIR-PLD and 193-nm-MAPLE depositions displayed the smoothest surfaces but did not show photoluminescence. (C) 2003...

  13. Characterization of ethylcellulose and hydroxypropyl methylcellulose thin films deposited by matrix-assisted pulsed laser evaporation

    Palla-Papavlu, A., E-mail: apalla@nipne.ro [National Institute for Lasers, Plasma and Radiation Physics, PO Box MG-36, Magurele, RO-077125 Bucharest (Romania); Rusen, L.; Dinca, V.; Filipescu, M. [National Institute for Lasers, Plasma and Radiation Physics, PO Box MG-36, Magurele, RO-077125 Bucharest (Romania); Lippert, T. [Paul Scherrer Institut, General Energy Research Department, 5232 Villigen PSI (Switzerland); Dinescu, M. [National Institute for Lasers, Plasma and Radiation Physics, PO Box MG-36, Magurele, RO-077125 Bucharest (Romania)

    2014-05-01

    In this study is reported the deposition of hydroxypropyl methylcellulose (HPMC) and ethylcellulose (EC) by matrix-assisted pulsed laser evaporation (MAPLE). Both HPMC and EC were deposited on silicon substrates using a Nd:YAG laser (266 nm, 5 ns laser pulse and 10 Hz repetition rate) and then characterized by atomic force microscopy and Fourier transform infrared spectroscopy. It was found that for laser fluences up to 450 mJ/cm{sup 2} the structure of the deposited HPMC and EC polymer in the thin film resembles to the bulk. Morphological investigations reveal island features on the surface of the EC thin films, and pores onto the HPMC polymer films. The obtained results indicate that MAPLE may be an alternative technique for the fabrication of new systems with desired drug release profile.

  14. Characterization of ethylcellulose and hydroxypropyl methylcellulose thin films deposited by matrix-assisted pulsed laser evaporation

    Palla-Papavlu, A.; Rusen, L.; Dinca, V.; Filipescu, M.; Lippert, T.; Dinescu, M.

    2014-05-01

    In this study is reported the deposition of hydroxypropyl methylcellulose (HPMC) and ethylcellulose (EC) by matrix-assisted pulsed laser evaporation (MAPLE). Both HPMC and EC were deposited on silicon substrates using a Nd:YAG laser (266 nm, 5 ns laser pulse and 10 Hz repetition rate) and then characterized by atomic force microscopy and Fourier transform infrared spectroscopy. It was found that for laser fluences up to 450 mJ/cm2 the structure of the deposited HPMC and EC polymer in the thin film resembles to the bulk. Morphological investigations reveal island features on the surface of the EC thin films, and pores onto the HPMC polymer films. The obtained results indicate that MAPLE may be an alternative technique for the fabrication of new systems with desired drug release profile.

  15. Growth of thin fullerene films by matrix assisted pulsed laser evaporation

    Canulescu, Stela; Schou, Jørgen; Fæster, Søren

    C60 fullerene thin films of average thickness of more than 100 nm on silicon substrates can be produced in vacuum by matrix-assisted pulsed laser evaporation (MAPLE). A 355 nm Nd:YAG laser was directed onto a frozen target of anisole with a concentration of 0.67 wt% C60. At laser fluences below 1.......5 J/cm2 the dominant fraction of the film molecules are C60 transferred to the substrate without any fragmentation. For high fluences high-resolution SEM images of MAPLE deposited films reveal large circular features on the surface with high amount of material concentrated at edges. These features......, observed over a wide range of laser fluences, are caused by ejection of large matrix-fullerene liquid droplets into the gas-phase and subsequent deposition. At similar laser energies, but using an unfocused laser beam, MAPLE favours evaporation of matrix and organic molecules, resulting in films...

  16. Matrix Assisted Pulsed Laser Evaporation for growth of fullerene thin films

    Canulescu, Stela; Schou, Jørgen; Fæster Nielsen, Søren

    C60 fullerene thin films of average thickness of more than 100 nm can be produced in vacuum by matrix-assisted pulsed laser evaporation (MAPLE). A 355 nm Nd:YAG laser was directed onto a frozen target of anisole with a concentration of 0.67 wt% C60. At laser fluences below 1.5 J/cm2, a dominant...... fraction of the film molecules are C60 transferred to the substrate without any fragmentation. Highresolution SEM images of MAPLE deposited films reveal large circular droplets on the surface with high amount of material concentrated at edges (Fig. 1A). These features, observed over a wide range of laser...... fluences, are caused by ejection of large matrix-fullerene liquid droplets into the gas-phase and subsequent deposition. At similar laser energies, but using an unfocused laser beam, MAPLE favours evaporation of matrix and organic molecules, resulting in production of films with smooth surfaces and minimal...

  17. Layered double hydroxides/polymer thin films grown by matrix assisted pulsed laser evaporation

    Birjega, R.; Matei, A.; Mitu, B.; Ionita, M.D.; Filipescu, M.; Stokker-Cheregi, F.; Luculescu, C.; Dinescu, M. [National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Str., 77125 Bucharest–Magurele (Romania); Zavoianu, R.; Pavel, O.D. [University of Bucharest, Faculty of Chemistry, Department of Chemical Technology and Catalysis, 4-12 Regina Elisabeta Bd., Bucharest (Romania); Corobea, M.C. [National R. and S. Institute for Chemistry and Petrochemistry, ICECHIM, 202 Splaiul Independentei Str., CP-35-274, 060021, Bucharest (Romania)

    2013-09-30

    Due to their highly tunable properties, layered double hydroxides (LDHs) are an emerging class of the favorably layered crystals used for the preparation of multifunctional polymer/layered crystal nanocomposites. In contrast to cationic clay materials with negatively charge layers, LDHs are the only host lattices with positively charged layers (brucite-like), with interlayer exchangeable anions and intercalated water. In this work, the deposition of thin films of Mg and Al based LDH/polymers nanocomposites by laser techniques is reported. Matrix assisted pulsed laser evaporation was the method used for thin films deposition. The Mg–Al LDHs capability to act as a host for polymers and to produce hybrid LDH/polymer films has been investigated. Polyethylene glycol with different molecular mass compositions and ethylene glycol were used as polymers. The structure and surface morphology of the deposited LDH/polymers films were examined by X-ray diffraction, Fourier transform infra-red spectroscopy, atomic force microscopy and scanning electron microscopy. - Highlights: • Hybrid composites deposited by matrix assisted pulsed laser evaporation (MAPLE). • Mg–Al layered double hydroxides (LDH) and polyethylene glycol (PEG) are used. • Mixtures of PEG1450 and LDH were deposited by MAPLE. • Deposited thin films preserve the properties of the starting material. • The film wettability can be controlled by the amount of PEG.

  18. Functionalized porphyrin conjugate thin films deposited by matrix assisted pulsed laser evaporation

    Iordache, S. [University of Bucharest, 3Nano-SAE Research Center, PO Box MG-38, Bucharest-Magurele (Romania); Cristescu, R., E-mail: rodica.cristescu@inflpr.ro [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Popescu, A.C.; Popescu, C.E.; Dorcioman, G.; Mihailescu, I.N. [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Ciucu, A.A. [University of Bucharest, Faculty of Chemistry, Bucharest (Romania); Balan, A.; Stamatin, I. [University of Bucharest, 3Nano-SAE Research Center, PO Box MG-38, Bucharest-Magurele (Romania); Fagadar-Cosma, E. [Institute of Chemistry Timisoara of Romanian Academy, M. Viteazul Ave. 24, 300223-Timisoara (Romania); Chrisey, D.B. [Tulane University, Departments of Physics and Biomedical Engineering, New Orleans, LA 70118 (United States)

    2013-08-01

    We report on the deposition of nanostructured porphyrin-base, 5(4-carboxyphenyl)-10,15,20-tris(4-phenoxyphenyl)-porphyrin thin films by matrix assisted pulsed laser evaporation onto silicon substrates with screen-printed electrodes. AFM investigations have shown that at 400 mJ/cm{sup 2} fluence a topographical transition takes place from the platelet-like stacking porphyrin-based nanostructures in a perpendicular arrangement to a quasi-parallel one both relative to the substrate surface. Raman spectroscopy has shown that the chemical structure of the deposited thin films is preserved for fluences within the range of 200–300 mJ/cm{sup 2}. Cyclic voltammograms have demonstrated that the free porphyrin is appropriate as a single mediator for glucose in a specific case of screen-printed electrodes, suggesting potential for designing a new class of biosensors.

  19. Design challenges for matrix assisted pulsed laser evaporation and infrared resonant laser evaporation equipment

    Greer, James A.

    2011-11-01

    Since the development of the Matrix Assisted Pulsed Laser Evaporation (MAPLE) process by the Naval Research Laboratory (NRL) in the late 1990s, MAPLE has become an active area of research for the deposition of a variety of polymer, biological, and organic thin films. As is often the case with advancements in thin-film deposition techniques new technology sometimes evolves by making minor or major adjustments to existing deposition process equipment and techniques. This is usually the quickest and least expensive way to try out new ideas and to "push the envelope" in order to obtain new and unique scientific results as quickly as possible. This process of "tweaking" current equipment usually works to some degree, but once the new process is further refined overall designs for a new deposition tool based on the critical attributes of the new process typically help capitalize more fully on the all the salient features of the new and improved process. This certainly has been true for the MAPLE process. In fact the first MAPLE experiments the polymer/solvent matrix was mixed and poured into a copper holder held at LN2 temperature on a laboratory counter top. The holder was then quickly placed onto a LN2 cooled reservoir in a vacuum deposition chamber and placed in a vertical position on a LN2 cooled stage and pumped down as quickly as possible. If the sample was not placed into the chamber quickly enough the frozen matrix would melt and drip into the bottom of the chamber onto the chambers main gate valve making a bit of a mess. However, skilled and motivated scientists usually worked quickly enough to make this process work most of the time. The initial results from these experiments were encouraging and led to several publications which sparked considerable interest in this newly developed technique Clearly this approach provided the vision that MAPLE was a viable deposition process, but the equipment was not optimal for conducting MAPLE experiments on a regular basis

  20. Nanoparticle Thin Films for Gas Sensors Prepared by Matrix Assisted Pulsed Laser Evaporation

    Roberto Rella

    2009-04-01

    Full Text Available The matrix assisted pulsed laser evaporation (MAPLE technique has been used for the deposition of metal dioxide (TiO2, SnO2 nanoparticle thin films for gas sensor applications. For this purpose, colloidal metal dioxide nanoparticles were diluted in volatile solvents, the solution was frozen at the liquid nitrogen temperature and irradiated with a pulsed excimer laser. The dioxide nanoparticles were deposited on Si and Al2O3 substrates. A rather uniform distribution of TiO2 nanoparticles with an average size of about 10 nm and of SnO2 nanoparticles with an average size of about 3 nm was obtained, as demonstrated by high resolution scanning electron microscopy (SEM-FEG inspections. Gas-sensing devices based on the resistive transduction mechanism were fabricated by depositing the nanoparticle thin films onto suitable rough alumina substrates equipped with interdigitated electrical contacts and heating elements. Electrical characterization measurements were carried out in controlled environment. The results of the gas-sensing tests towards low concentrations of ethanol and acetone vapors are reported. Typical gas sensor parameters (gas responses, response/recovery time, sensitivity, and low detection limit towards ethanol and acetone are presented.

  1. Nanoparticle thin films for gas sensors prepared by matrix assisted pulsed laser evaporation.

    Caricato, Anna Paola; Luches, Armando; Rella, Roberto

    2009-01-01

    The matrix assisted pulsed laser evaporation (MAPLE) technique has been used for the deposition of metal dioxide (TiO(2), SnO(2)) nanoparticle thin films for gas sensor applications. For this purpose, colloidal metal dioxide nanoparticles were diluted in volatile solvents, the solution was frozen at the liquid nitrogen temperature and irradiated with a pulsed excimer laser. The dioxide nanoparticles were deposited on Si and Al(2)O(3) substrates. A rather uniform distribution of TiO(2) nanoparticles with an average size of about 10 nm and of SnO(2) nanoparticles with an average size of about 3 nm was obtained, as demonstrated by high resolution scanning electron microscopy (SEM-FEG) inspections. Gas-sensing devices based on the resistive transduction mechanism were fabricated by depositing the nanoparticle thin films onto suitable rough alumina substrates equipped with interdigitated electrical contacts and heating elements. Electrical characterization measurements were carried out in controlled environment. The results of the gas-sensing tests towards low concentrations of ethanol and acetone vapors are reported. Typical gas sensor parameters (gas responses, response/recovery time, sensitivity, and low detection limit) towards ethanol and acetone are presented.

  2. Protein-resistant polymer coatings obtained by matrix assisted pulsed laser evaporation

    Rusen, L. [National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, 077125, Magurele, Bucharest (Romania); Mustaciosu, C. [Horia Hulubei National Institute of Physics and Nuclear Engineering - IFIN HH, Magurele, Bucharest (Romania); Mitu, B.; Filipescu, M.; Dinescu, M. [National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, 077125, Magurele, Bucharest (Romania); Dinca, V., E-mail: dinali@nipne.ro [National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, 077125, Magurele, Bucharest (Romania)

    2013-08-01

    Adsorption of proteins and polysaccharides is known to facilitate microbial attachment and subsequent formation of biofilm on surfaces that ultimately results in its biofouling. Therefore, protein repellent modified surfaces are necessary to block the irreversible attachment of microorganisms. Within this context, the feasibility of using the Poly(ethylene glycol)-block-poly(ε-caprolactone) methyl ether (PEG-block-PCL Me) copolymer as potential protein-resistant coating was explored in this work. The films were deposited using Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique that allows good control of composition, thickness and homogeneity. The chemical and morphological characteristics of the films were examined using Fourier Transform Infrared Spectroscopy (FTIR), contact angle measurements and Atomic Force Microscopy (AFM). The FTIR data demonstrates that the functional groups in the MAPLE-deposited films remain intact, especially for fluences below 0.5 J cm{sup −2}. Optical Microscopy and AFM images show that the homogeneity and the roughness of the coatings are related to both laser parameters (fluence, number of pulses) and target composition. Protein adsorption tests were performed on the PEG-block-PCL Me copolymer coated glass and on bare glass surface as a control. The results show that the presence of copolymer as coating significantly reduces the adsorption of proteins.

  3. Surface morphology of polyethylene glycol films produced by matrix-assisted pulsed laser evaporation (MAPLE): Dependence on substrate temperature

    Rodrigo, K.; Czuba, P.; Toftmann, B.

    2006-01-01

    The dependence of the surface morphology on the substrate temperature during film deposition was investigated for polyethylene glycol (PEG) films by matrix-assisted pulsed laser evaporation (MAPLE). The surface structure was studied with a combined technique of optical imaging and AFM measurements...

  4. Deposition of Methylammonium Lead Triiodide by Resonant Infrared Matrix-Assisted Pulsed Laser Evaporation

    Barraza, E. Tomas; Dunlap-Shohl, Wiley A.; Mitzi, David B.; Stiff-Roberts, Adrienne D.

    2018-02-01

    Resonant infrared matrix-assisted pulsed laser evaporation (RIR-MAPLE) was used to deposit the metal-halide perovskite (MHP) CH3NH3PbI3 (methylammonium lead triiodide, or MAPbI), creating phase-pure films. Given the moisture sensitivity of these crystalline, multi-component organic-inorganic hybrid materials, deposition of MAPbI by RIR-MAPLE required a departure from the use of water-based emulsions as deposition targets. Different chemistries were explored to create targets that properly dissolved MAPbI components, were stable under vacuum conditions, and enabled resonant laser energy absorption. Secondary phases and solvent contamination in the resulting films were studied through Fourier transform infrared (FTIR) absorbance and x-ray diffraction (XRD) measurements, suggesting that lingering excess methylammonium iodide (MAI) and low-vapor pressure solvents can distort the microstructure, creating crystalline and amorphous non-perovskite phases. Thermal annealing of films deposited by RIR-MAPLE allowed for excess solvent to be evaporated from films without degrading the MAPbI structure. Further, it was demonstrated that RIR-MAPLE does not require excess MAI to create stoichiometric films with optoelectronic properties, crystal structure, and film morphology comparable to films created using more established spin-coating methods for processing MHPs. This work marks the first time a MAPLE-related technique was used to deposit MHPs.

  5. Biomimetic nanocrystalline apatite coatings synthesized by Matrix Assisted Pulsed Laser Evaporation for medical applications

    Visan, A. [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Grossin, D. [CIRIMAT – Carnot Institute, University of Toulouse, ENSIACET, 4 Allée Emile Monso, 31030 Toulouse Cedex 4 (France); Stefan, N.; Duta, L.; Miroiu, F.M. [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Stan, G.E. [National Institute of Materials Physics, RO-077125, Magurele-Ilfov (Romania); Sopronyi, M.; Luculescu, C. [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Freche, M.; Marsan, O.; Charvilat, C. [CIRIMAT – Carnot Institute, University of Toulouse, ENSIACET, 4 Allée Emile Monso, 31030 Toulouse Cedex 4 (France); Ciuca, S. [Politehnica University of Bucharest, Faculty of Materials Science and Engineering, Bucharest (Romania); Mihailescu, I.N., E-mail: ion.mihailescu@inflpr.ro [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania)

    2014-02-15

    Highlights: • We report the deposition by MAPLE of biomimetic apatite coatings on Ti substrates. • This is the first report of MAPLE deposition of hydrated biomimetic apatite films. • Biomimetic apatite powder was synthesized by double decomposition process. • Non-apatitic environments, of high surface reactivity, are preserved post-deposition. • We got the MAPLE complete transfer as thin film of a hydrated, delicate material. -- Abstract: We report the deposition by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique of biomimetic nanocrystalline apatite coatings on titanium substrates, with potential application in tissue engineering. The targets were prepared from metastable, nanometric, poorly crystalline apatite powders, analogous to mineral bone, synthesized through a biomimetic approach by double decomposition process. For the deposition of thin films, a KrF* excimer laser source was used (λ = 248 nm, τ{sub FWHM} ≤ 25 ns). The analyses revealed the existence, in synthesized powders, of labile non-apatitic mineral ions, associated with the formation of a hydrated layer at the surface of the nanocrystals. The thin film analyses showed that the structural and chemical nature of the nanocrystalline apatite was prevalently preserved. The perpetuation of the non-apatitic environments was also observed. The study indicated that MAPLE is a suitable technique for the congruent transfer of a delicate material, such as the biomimetic hydrated nanohydroxyapatite.

  6. Biomimetic nanocrystalline apatite coatings synthesized by Matrix Assisted Pulsed Laser Evaporation for medical applications

    Visan, A.; Grossin, D.; Stefan, N.; Duta, L.; Miroiu, F.M.; Stan, G.E.; Sopronyi, M.; Luculescu, C.; Freche, M.; Marsan, O.; Charvilat, C.; Ciuca, S.; Mihailescu, I.N.

    2014-01-01

    Highlights: • We report the deposition by MAPLE of biomimetic apatite coatings on Ti substrates. • This is the first report of MAPLE deposition of hydrated biomimetic apatite films. • Biomimetic apatite powder was synthesized by double decomposition process. • Non-apatitic environments, of high surface reactivity, are preserved post-deposition. • We got the MAPLE complete transfer as thin film of a hydrated, delicate material. -- Abstract: We report the deposition by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique of biomimetic nanocrystalline apatite coatings on titanium substrates, with potential application in tissue engineering. The targets were prepared from metastable, nanometric, poorly crystalline apatite powders, analogous to mineral bone, synthesized through a biomimetic approach by double decomposition process. For the deposition of thin films, a KrF* excimer laser source was used (λ = 248 nm, τ FWHM ≤ 25 ns). The analyses revealed the existence, in synthesized powders, of labile non-apatitic mineral ions, associated with the formation of a hydrated layer at the surface of the nanocrystals. The thin film analyses showed that the structural and chemical nature of the nanocrystalline apatite was prevalently preserved. The perpetuation of the non-apatitic environments was also observed. The study indicated that MAPLE is a suitable technique for the congruent transfer of a delicate material, such as the biomimetic hydrated nanohydroxyapatite

  7. Organic/hybrid thin films deposited by matrix-assisted pulsed laser evaporation (MAPLE)

    Stiff-Roberts, Adrienne D.; Ge, Wangyao

    2017-12-01

    Some of the most exciting materials research in the 21st century attempts to resolve the challenge of simulating, synthesizing, and characterizing new materials with unique properties designed from first principles. Achievements in such development for organic and organic-inorganic hybrid materials make them important options for electronic and/or photonic devices because they can impart multi-functionality, flexibility, transparency, and sustainability to emerging systems, such as wearable electronics. Functional organic materials include small molecules, oligomers, and polymers, while hybrid materials include inorganic nanomaterials (such as zero-dimensional quantum dots, one-dimensional carbon nanotubes, or two-dimensional nanosheets) combined with organic matrices. A critically important step to implementing new electronic and photonic devices using such materials is the processing of thin films. While solution-based processing is the most common laboratory technique for organic and hybrid materials, vacuum-based deposition has been critical to the commercialization of organic light emitting diodes based on small molecules, for example. Therefore, it is desirable to explore vacuum-based deposition of organic and hybrid materials that include larger macromolecules, such as polymers. This review article motivates the need for physical vapor deposition of polymeric and hybrid thin films using matrix-assisted pulsed laser evaporation (MAPLE), which is a type of pulsed laser deposition. This review describes the development of variations in the MAPLE technique, discusses the current understanding of laser-target interactions and growth mechanisms for different MAPLE variations, surveys demonstrations of MAPLE-deposited organic and hybrid materials for electronic and photonic devices, and provides a future outlook for the technique.

  8. High fluence deposition of polyethylene glycol films at 1064 nm by matrix assisted pulsed laser evaporation (MAPLE)

    Purice, Andreea; Schou, Jørgen; Kingshott, P.

    2007-01-01

    Matrix assisted pulsed laser evaporation (MAPLE) has been applied for deposition of thin polyethylene glycol (PEG) films with infrared laser light at 1064 nm. We have irradiated frozen targets (of 1 wt.% PEG dissolved in water) and measured the deposition rate in situ with a quartz crystal 2...... microbalance. The laser fluence needed to produce PEG films turned out to be unexpectedly high with a threshold of 9 J/cm(2) and the deposition rate was much lower than that with laser light at 355 nm. Results from matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI......-TOF-MS) analysis demonstrate that the chemistry, molecular weight and polydispersity of the PEG films were identical to the starting material. Studies of the film surface with scanning electron microscopy (SEM) indicate that the Si-substrate is covered by a relatively homogenous PEG film with few bare spots. (c...

  9. The influence of wavelength, temporal sequencing, and pulse duration on resonant infrared matrix-assisted laser processing of polymer films

    O'Malley, S. M.; Schoeffling, Jonathan; Jimenez, Richard; Zinderman, Brian; Yi, SunYong; Bubb, D. M.

    2014-06-01

    We have carried out a systematic investigation of laser ablation plume interactions in resonant infrared matrix-assisted pulsed laser evaporation. The laser source utilized in this study was a mid-infrared OPO capable of dual sequential ns pulses with adjustable delay ranging from 1 to 100 μs. This unique capability enabled us both to probe the ablation plume with a second laser pulse, and to effectively double the laser fluence. The primary ablation target used for this study consisted of poly(methyl methacrylate) dissolved in a binary mixture of methanol and toluene. Both the critical thermodynamic and optical properties of the binary mixture were determined and used to interpret our results. We found that deposition rates associated with single pulse irradiation tracks with the optical absorption coefficient in the spectral range from 2,700 to 3,800 nm. In the case of dual sequential pulses, discrepancies in this trend have been linked to the rate of change in the optical absorption coefficient with temperature. The influence of fluence on deposition rate was found to follow a sigmoidal dependence. Surface roughness was observed to have a diametrically opposed trend with pulse delay depending on whether the OH or CH vibrational mode was excited. In the case of CH excitation, we suggest that the rougher films are due to the absorbance of the second pulse by droplets within the plume containing residual solvent which leads to the formation of molecular balloons and hence irregularly shaped features on the substrate.

  10. Study of titania nanorod films deposited by matrix-assisted pulsed laser evaporation as a function of laser fluence

    Caricato, A. P.; Belviso, M. R.; Catalano, M.; Cesaria, M.; Cozzoli, P. D.; Luches, A.; Manera, M. G.; Martino, M.; Rella, R.; Taurino, A.

    2011-11-01

    Chemically synthesized brookite titanium dioxide (TiO2) nanorods with average diameter and length dimensions of 3-4 nm and 35-50 nm, respectively, were deposited by the matrix-assisted pulsed laser evaporation technique. A toluene nanorod solution was frozen at the liquid-nitrogen temperature and irradiated with a KrF excimer laser ( λ=248 nm, τ=20 ns) at the repetition rate of 10 Hz, at different fluences (25 to 350 mJ/cm2). The deposited films were structurally characterized by high-resolution scanning and transmission electron microscopy. single-crystal Si wafers and carbon-coated Cu grids were used as substrates. Structural analyses evidenced the occurrence of brookite-phase crystalline nanospheres coexisting with individually distinguishable TiO2 nanorods in the films deposited at fluences varying from 50 to 350 mJ/cm2. Nanostructured TiO2 films comprising only nanorods were deposited by lowering the laser fluence to 25 mJ/cm2. The observed shape and phase transitions of the nanorods are discussed taking into account the laser-induced heating effects, reduced melting temperature and size-dependent thermodynamic stability of nanoscale TiO2.

  11. Uniform thin films of TiO2 nanoparticles deposited by matrix-assisted pulsed laser evaporation

    Caricato, A.P.; Manera, M.G.; Martino, M.; Rella, R.; Romano, F.; Spadavecchia, J.; Tunno, T.; Valerini, D.

    2007-01-01

    We report morphological and optical properties of a colloidal TiO 2 nanoparticle film, deposited on a quartz substrate by using the Matrix-Assisted Pulsed Laser Evaporation (MAPLE) technique. Atomic Force Microscopy demonstrated that a good uniformity of the deposition can be obtained. The presence of agglomerates with dimensions of about 1 μm in size was noticed. Form UV-vis transmission spectra, recorded in the 200-800 nm range, the optical constants and the energy gap were determined besides the film thickness. The optical constants resulted in agreement with the values reported in literature for TiO 2 nanoparticle thin films

  12. Deposition of matrix-free fullerene films with improved morphology by matrix-assisted pulsed laser evaporation (MAPLE)

    Canulescu, Stela; Schou, Jørgen; Fæster, Søren

    2013-01-01

    Thin films of C60 were deposited by matrix-assisted pulsed laser evaporation (MAPLE) from a frozen target of anisole with 0.67 wt% C60. Above a fluence of 1.5 J/cm2 the C60 films are strongly non-uniform and are resulting from transfer of matrix-droplets containing fullerenes. At low fluence...... the fullerene molecules in the films are intact, the surface morphology is substantially improved and there are no measurable traces of the matrix molecules in the film. This may indicate a regime of dominant evaporation at low fluence which merges into the MAPLE regime of liquid ejection of the host matrix...

  13. Temperature responsive functional polymeric thin films obtained by matrix assisted pulsed laser evaporation for cells attachment–detachment study

    Rusen, L. [NILPRP, National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, RO-077125 Magurele, Bucharest (Romania); Dinca, V., E-mail: dinali@nipne.ro [NILPRP, National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, RO-077125 Magurele, Bucharest (Romania); Mitu, B. [NILPRP, National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, RO-077125 Magurele, Bucharest (Romania); Mustaciosu, C. [Horia Hulubei National Institute of Physics and Nuclear Engineering, IFIN HH, Magurele, Bucharest (Romania); Dinescu, M. [NILPRP, National Institute for Lasers, Plasma and Radiation Physics, 409 Atomistilor Street, PO Box MG-16, RO-077125 Magurele, Bucharest (Romania)

    2014-05-01

    Multifunctional thin films used as thermoresponsive substrate for engineering cell sheets represent an important area in tissue engineering. As the morphology and the chemical characteristics of the thin films directly control their interaction with cells, it is important to correlate these characteristics with the biological answer. In this study, thermally sensitive poly(N-isopropylacrylamide), (pNIPAAm) thin films were prepared by matrix assisted pulsed laser evaporation and utilized in L929 cell adhesion and detachment studies. Fourier transform infrared spectroscopy (FTIR) and atomic force microscopy (AFM) were used to determine the pNIPAAm thin films chemical and morphological characteristics. The FTIR data demonstrated that the functional groups in the MAPLE-deposited films remained intact for fluences in the range of 200–600 mJ cm{sup −2}. Within this fluence range, the AFM topographical studies showed that the roughness of the coatings was dependent on laser fluence and the obtained surfaces were characterized by a granular aspect. L929 cell viability studies onto the pNIPAAm coatings showed little or no toxic effect for fluences below 600 mJ cm{sup −2}, while for higher fluences, viability was decreased with more than 50%. The adhesion and detachment of the cell was found to be mainly dependent on the film surface morphology.

  14. Photoluminescence of Eu-doped LiYF4 thin films grown by pulsed laser deposition and matrix-assisted pulsed laser evaporation

    Stokker-Cheregi, F; Matei, A; Dinescu, M; Secu, C E; Secu, M

    2014-01-01

    Matrix-assisted pulsed laser evaporation (MAPLE) has been investigated as an alternative to the pulsed laser deposition (PLD) technique for Eu 3+ -doped crystalline LiYF 4 thin-films deposition. MAPLE assumes laser ablation of a frozen target made of the material of interest diluted in a solvent, rather than that of a bulk target, of either pressed powder or single crystal, used in the case of PLD. Our approach stems from the assumption that laser ablation of a frozen dilute target would result in thin films with improved morphology, as compared to PLD. Indeed, we find that roughness values of samples obtained by the MAPLE technique are four times lower than in the case of PLD. A lower transmittance was noticed for PLD obtained layers with respect to those grown by MAPLE due to strong scattering of light by the morphological defects. Photoluminescence spectra are showing characteristic Eu 3+ -ion luminescence bands at 578, 591, 612, 650 and 698 nm ( 5 D 0  →  7 F J ); crystal field splitting of the bands indicates dopant ions incorporation in the host material during transfer by either PLD or MAPLE. (paper)

  15. Resonant Infrared Matrix-Assisted Pulsed Laser Evaporation Of Inorganic Nanoparticles And Organic/Inorganic Hybrid Nanocomposites

    Pate, Ryan; Lantz, Kevin R.; Dhawan, Anuj; Vo-Dinh, Tuan; Stiff-Roberts, Adrienne D.

    2010-10-01

    In this research, resonant infrared matrix-assisted pulsed laser evaporation (RIR-MAPLE) has been used to deposit different classes of inorganic nanoparticles, including bare, un-encapsulated ZnO and Au nanoparticles, as well as ligand-encapsulated CdSe colloidal quantum dots (CQDs). RIR-MAPLE has been used for thin-film deposition of different organic/inorganic hybrid nanocomposites using some of these inorganic nanoparticles, including CdSe CQD-poly[2-methoxy-5-(2'-ethylhexyloxy)-1,4-(1-cyanovinylene)phenylene] (MEH-CN-PPV) nanocomposites and Au nanoparticle-poly(methyl methacrylate) (PMMA) nanocomposites. The unique contribution of this research is that a technique is demonstrated for the deposition of organic-based thin-films requiring solvents with bond energies that do not have to be resonant with the laser energy. By creating an emulsion of solvent and ice in the target, RIR-MAPLE using a 2.94 μm laser can deposit most material systems because the hydroxyl bonds in the ice component of the emulsion matrix are strongly resonant with the 2.94 μm laser. In this way, the types of materials that can be deposited using RIR-MAPLE has been significantly expanded. Furthermore, materials with different solvent bond energies can be co-deposited without concern for material degradation and without the need to specifically tune the laser energy to each material solvent bond energy, thereby facilitating the realization of organic/inorganic hybrid nanocomposite thin-films. In addition to the structural characterization of the inorganic nanoparticle and hybrid nanocomposite thin-films deposited using this RIR-MAPLE technique, optical characterization is presented to demonstrate the potential of such films for optoelectronic device applications.

  16. Resonant Infrared Matrix-Assisted Pulsed Laser Evaporation Of Inorganic Nanoparticles And Organic/Inorganic Hybrid Nanocomposites

    Pate, Ryan; Lantz, Kevin R.; Stiff-Roberts, Adrienne D.; Dhawan, Anuj; Vo-Dinh, Tuan

    2010-01-01

    In this research, resonant infrared matrix-assisted pulsed laser evaporation (RIR-MAPLE) has been used to deposit different classes of inorganic nanoparticles, including bare, un-encapsulated ZnO and Au nanoparticles, as well as ligand-encapsulated CdSe colloidal quantum dots (CQDs). RIR-MAPLE has been used for thin-film deposition of different organic/inorganic hybrid nanocomposites using some of these inorganic nanoparticles, including CdSe CQD-poly[2-methoxy-5-(2'-ethylhexyloxy )-1,4-(1-cyanovinylene)phenylene](MEH-CN-PPV) nanocomposites and Au nanoparticle-poly(methyl methacrylate)(PMMA) nanocomposites. The unique contribution of this research is that a technique is demonstrated for the deposition of organic-based thin-films requiring solvents with bond energies that do not have to be resonant with the laser energy. By creating an emulsion of solvent and ice in the target, RIR-MAPLE using a 2.94 μm laser can deposit most material systems because the hydroxyl bonds in the ice component of the emulsion matrix are strongly resonant with the 2.94 μm laser. In this way, the types of materials that can be deposited using RIR-MAPLE has been significantly expanded. Furthermore, materials with different solvent bond energies can be co-deposited without concern for material degradation and without the need to specifically tune the laser energy to each material solvent bond energy, thereby facilitating the realization of organic/inorganic hybrid nanocomposite thin-films. In addition to the structural characterization of the inorganic nanoparticle and hybrid nanocomposite thin-films deposited using this RIR-MAPLE technique, optical characterization is presented to demonstrate the potential of such films for optoelectronic device applications.

  17. Deposition of organic dyes for dye-sensitized solar cell by using matrix-assisted pulsed laser evaporation

    Chih-Ping Yen

    2016-08-01

    Full Text Available The deposition of various distinct organic dyes, including ruthenium complex N3, melanin nanoparticle (MNP, and porphyrin-based donor-π-acceptor dye YD2-o-C8, by using matrix-assisted pulsed laser evaporation (MAPLE for application to dye-sensitized solar cell (DSSC is investigated systematically. It is found that the two covalently-bonded organic molecules, i.e., MNP and YD2-o-C8, can be transferred from the frozen target to the substrate with maintained molecular integrity. In contrast, N3 disintegrates in the process, presumably due to the lower bonding strength of metal complex compared to covalent bond. With the method, DSSC using YD2-o-C8 is fabricated, and an energy conversion efficiency of 1.47% is attained. The issue of the low penetration depth of dyes deposited by MAPLE and the possible resolution to it are studied. This work demonstrates that MAPLE could be an alternative way for deposition of organic dyes for DSSC.

  18. Deposition of organic dyes for dye-sensitized solar cell by using matrix-assisted pulsed laser evaporation

    Yen, Chih-Ping [Department of Physics, National Taiwan University, Taipei 106, Taiwan (China); Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan (China); Yu, Pin-Feng [Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan (China); Department of Physics, National Chung Cheng University, Chiayi 621, Taiwan (China); Wang, Jyhpyng [Department of Physics, National Taiwan University, Taipei 106, Taiwan (China); Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan (China); Department of Physics, National Central University, Taoyuan 320, Taiwan (China); Lin, Jiunn-Yuan [Department of Physics, National Chung Cheng University, Chiayi 621, Taiwan (China); Chen, Yen-Mu [SuperbIN Co., Ltd., Taipei 114, Taiwan (China); Chen, Szu-yuan, E-mail: sychen@ltl.iams.sinica.edu.tw [Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan (China); Department of Physics, National Central University, Taoyuan 320, Taiwan (China)

    2016-08-15

    The deposition of various distinct organic dyes, including ruthenium complex N3, melanin nanoparticle (MNP), and porphyrin-based donor-π-acceptor dye YD2-o-C8, by using matrix-assisted pulsed laser evaporation (MAPLE) for application to dye-sensitized solar cell (DSSC) is investigated systematically. It is found that the two covalently-bonded organic molecules, i.e., MNP and YD2-o-C8, can be transferred from the frozen target to the substrate with maintained molecular integrity. In contrast, N3 disintegrates in the process, presumably due to the lower bonding strength of metal complex compared to covalent bond. With the method, DSSC using YD2-o-C8 is fabricated, and an energy conversion efficiency of 1.47% is attained. The issue of the low penetration depth of dyes deposited by MAPLE and the possible resolution to it are studied. This work demonstrates that MAPLE could be an alternative way for deposition of organic dyes for DSSC.

  19. Thin films of polymer blends deposited by matrix-assisted pulsed laser evaporation: Effects of blending ratios

    Paun, Irina Alexandra; Ion, Valentin; Moldovan, Antoniu; Dinescu, Maria

    2011-01-01

    In this work, we show successful use of matrix-assisted pulsed laser evaporation (MAPLE) for obtaining thin films of PEG:PLGA blends, in the view of their use for controlled drug delivery. In particular, we investigate the influence of the blending ratios on the characteristics of the films. We show that the roughness of the polymeric films is affected by the ratio of each polymer within the blend. In addition, we perform Fourier transformed infrared spectroscopy (FTIR) measurements and we find that the intensities ratios of the infrared absorption bands of the two polymers are consistent with the blending ratios. Finally, we assess the optical constants of the polymeric films by spectroscopic ellipsometry (SE). We point out that the blending ratios exert an influence on the optical characteristics of the films and we validate the SE results by atomic force microscopy and UV-vis spectrophotometry. In all, we stress that the ratios in which the two polymers are blended have significant impact on the morphology, chemical structure and optical characteristics of the polymeric films deposited by MAPLE.

  20. Composite biocompatible hydroxyapatite-silk fibroin coatings for medical implants obtained by Matrix Assisted Pulsed Laser Evaporation

    Miroiu, F.M.; Socol, G.; Visan, A.; Stefan, N.; Craciun, D.; Craciun, V.; Dorcioman, G.; Mihailescu, I.N.; Sima, L.E.; Petrescu, S.M.; Andronie, A.; Stamatin, I.; Moga, S.; Ducu, C.

    2010-01-01

    The aim of this study was to obtain biomimetic inorganic-organic thin films as coatings for metallic medical implants. These contain hydroxyapatite, the inorganic component of the bony tissues, and a natural biopolymer - silk fibroin - added in view to induce the surface functionalization. Hydroxyapatite (HA), silk fibroin (FIB) and composite HA-FIB films were obtained by Matrix Assisted Pulsed Laser Evaporation (MAPLE) in order to compare their physical and biological performances as coatings on metallic prostheses. We used an excimer laser source (KrF*, λ = 248 nm, τ = 25 ns) operated at 10 Hz repetition rate. Coatings were deposited on quartz, Si and Ti substrates and then subjected to physical (FTIR, XRD, AFM, SEM) analyses, correlated with the results of the cytocompatibility in vitro tests. The hybrid films were synthesized from frozen targets of aqueous suspensions with 3:2 or 3:4 weight ratio of HA:FIB. An appropriate stoichiometric and functional transfer was obtained for 0.4-0.5 J/cm 2 laser fluence. FTIR spectra of FIB and HA-FIB films exhibited distinctive absorption maxima, in specific positions of FIB random coil form: 1540 cm -1 amide II, 1654 cm -1 amide I, 1243 cm -1 amide III, while the peak from 1027 cm -1 appeared only for HA and composite films. Osteosarcoma SaOs2 cells cultured 72 h on FIB and HA-FIB films showed increased viability, good spreading and normal cell morphology. The well-elongated, flattened cells are a sign of an appropriate interaction with the MAPLE FIB and composite HA-FIB coatings.

  1. Composite biocompatible hydroxyapatite-silk fibroin coatings for medical implants obtained by Matrix Assisted Pulsed Laser Evaporation

    Miroiu, F.M., E-mail: marimona.miroiu@inflpr.ro [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Socol, G.; Visan, A.; Stefan, N.; Craciun, D.; Craciun, V.; Dorcioman, G.; Mihailescu, I.N. [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Sima, L.E.; Petrescu, S.M. [Institute of Biochemistry, Romanian Academy, 296 Splaiul Independentei, 060031 Bucharest (Romania); Andronie, A.; Stamatin, I. [3Nano-SAE Alternative Energy Sources-University of Bucharest, Faculty of Physics, 409 Atomistilor Street, RO-77125, Magurele-Ilfov (Romania); Moga, S.; Ducu, C. [University of Pitesti, Targul din Vale Str, no. 1, 110040 Pitesti (Romania)

    2010-05-25

    The aim of this study was to obtain biomimetic inorganic-organic thin films as coatings for metallic medical implants. These contain hydroxyapatite, the inorganic component of the bony tissues, and a natural biopolymer - silk fibroin - added in view to induce the surface functionalization. Hydroxyapatite (HA), silk fibroin (FIB) and composite HA-FIB films were obtained by Matrix Assisted Pulsed Laser Evaporation (MAPLE) in order to compare their physical and biological performances as coatings on metallic prostheses. We used an excimer laser source (KrF*, {lambda} = 248 nm, {tau} = 25 ns) operated at 10 Hz repetition rate. Coatings were deposited on quartz, Si and Ti substrates and then subjected to physical (FTIR, XRD, AFM, SEM) analyses, correlated with the results of the cytocompatibility in vitro tests. The hybrid films were synthesized from frozen targets of aqueous suspensions with 3:2 or 3:4 weight ratio of HA:FIB. An appropriate stoichiometric and functional transfer was obtained for 0.4-0.5 J/cm{sup 2} laser fluence. FTIR spectra of FIB and HA-FIB films exhibited distinctive absorption maxima, in specific positions of FIB random coil form: 1540 cm{sup -1} amide II, 1654 cm{sup -1} amide I, 1243 cm{sup -1} amide III, while the peak from 1027 cm{sup -1} appeared only for HA and composite films. Osteosarcoma SaOs2 cells cultured 72 h on FIB and HA-FIB films showed increased viability, good spreading and normal cell morphology. The well-elongated, flattened cells are a sign of an appropriate interaction with the MAPLE FIB and composite HA-FIB coatings.

  2. Nanoparticle and nanorod films deposited by matrix assisted pulsed laser evaporation

    Caricato, A. P.; Cesaria, M.; Luches, A.; Martino, M.

    2012-07-01

    The promising results obtained with the MAPLE-deposition of nanostructured thin films, to be used in different fields, are reviewed. Nanoparticles (TiO2, SnO2, CdS) and nanorods (TiO2) with well defined dimensions were suspended in appropriate solvents (distilled water, toluene) with low concentration (1wt% or less). The solutions were flash frozen at the liquid nitrogen temperature to form the targets to be laser irradiated. The MAPLE process allowed a successful transfer from the target to rough and flat substrates, preserving the starting composition and crystalline phase of the nanostructures in a wide range of experimental conditions. In contrast, a careful choice of the laser fluence is mandatory to avoid shape modifications. Growth of metal nanoparticles with a low dispersion in size was also obtained by the MAPLE technique, starting from target solutions of a metallorganic element (AcPd) diluted in different solvents (acetone, diethyl ether). It seems that selecting the solvent with appropriate values of viscosity and boiling temperatures, it is possible to modulate the nanoparticles size. Most of the deposited nanostructured films were tested as sensing elements for gas sensors.

  3. Nonlinear optical studies on 4-(ferrocenylmethylimino)-2-hydroxy-benzoic acid thin films deposited by matrix-assisted pulsed laser evaporation (MAPLE)

    Matei, Andreea [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); Marinescu, Maria, E-mail: maria.marinescu@chimie.unibuc.ro [UB - University of Bucharest, Faculty of Chemistry, 90-92 Şoseaua Panduri, Sector 5, RO-010184, Bucharest (Romania); Constantinescu, Catalin, E-mail: catalin.constantinescu@inflpr.ro [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); Ion, Valentin; Mitu, Bogdana [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); Ionita, Iulian [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); UB - University of Bucharest, Faculty of Physics, 405 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); Dinescu, Maria [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); Emandi, Ana [INFLPR - National Institute for Laser, Plasma and Radiation Physics, 409 Atomistilor Str., Magurele RO-077125, Bucharest (Romania); UB - University of Bucharest, Faculty of Chemistry, 90-92 Şoseaua Panduri, Sector 5, RO-010184, Bucharest (Romania)

    2016-06-30

    Graphical abstract: - Highlights: • A newly synthesized ferrocene-derivative exhibits SHG potential. • Matrix-assisted pulsed laser evaporation is employed for thin film fabrication. • The optical properties of the films are investigated, presented and discussed. • At maximum laser output power, the SHG signal is strongly influenced by thin film thickness. - Abstract: We present results on a new, laboratory synthesized ferrocene-derivative, i.e. 4-(ferrocenylmethylimino)-2-hydroxy-benzoic acid. Thin films with controlled thickness are deposited by matrix-assisted pulsed laser evaporation (MAPLE), on quartz and silicon substrates, with the aim of evaluating the nonlinear optical properties for potential optoelectronic applications. Dimethyl sulfoxide was used as matrix, with 1% wt. concentration of the guest compound. The frozen target is irradiated by using a Nd:YAG laser (4ω/266 nm, 7 ns pulse duration, 10 Hz repetition rate), at low fluences ranging from 0.1 to 1 J/cm{sup 2}. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) are used to probe the surface morphology of the films. Fourier transform infrared (FTIR) and Raman spectroscopy reveal similar structure of the thin film material when compared to the starting material. The optical properties of the thin films are investigated by spectroscopic-ellipsometry (SE), and the refractive index dependence with respect to temperature is studied. The second harmonic generation (SHG) potential is assessed by using a femtosecond Ti:sapphire laser (800 nm, 60–100 fs pulse duration, 80 MHz repetition rate), at 200 mW maximum output power, revealing that the SHG signal intensity is strongly influenced by the films’ thickness.

  4. Study of temperature dependence and angular distribution of poly(9,9-dioctylfluorene) polymer films deposited by matrix-assisted pulsed laser evaporation (MAPLE)

    Caricato, A.P.; Anni, M.; Manera, M.G.; Martino, M.; Rella, R.; Romano, F.; Tunno, T.; Valerini, D.

    2009-01-01

    Poly(9,9-dioctylfluorene) (PFO) polymer films were deposited by matrix-assisted pulsed laser evaporation (MAPLE) technique. The polymer was diluted (0.5 wt%) in tetrahydrofuran and, once cooled to liquid nitrogen temperature, it was irradiated with a KrF excimer laser. 10,000 laser pulses were used to deposit PFO films on Si substrates at different temperatures (-16, 30, 50 and 70 deg. C). One PFO film was deposited with 16,000 laser pulses at a substrate temperature of 50 deg. C. The morphology, optical and structural properties of the films were investigated by SEM, AFM, PL and FTIR spectroscopy. SEM inspection showed different characteristic features on the film surface, like deflated balloons, droplets and entangled polymer filaments. The roughness of the films was, at least partially, controlled by substrate heating, which however had the effect to reduce the deposition rate. The increase of the laser pulse number modified the target composition and increased the surface roughness. The angular distribution of the material ejected from the target confirmed the forward ejection of the target material. PFO films presented negligible modification of the chemical structure respect to the bulk material.

  5. Organic Thin Films Deposited by Emulsion-Based, Resonant Infrared, Matrix-Assisted Pulsed Laser Evaporation: Fundamentals and Applications

    Ge, Wangyao

    Thin film deposition techniques are indispensable to the development of modern technologies as thin film based optical coatings, optoelectronic devices, sensors, and biological implants are the building blocks of many complicated technologies, and their performance heavily depends on the applied deposition technique. Particularly, the emergence of novel solution-processed materials, such as soft organic molecules, inorganic compounds and colloidal nanoparticles, facilitates the development of flexible and printed electronics that are inexpensive, light weight, green and smart, and these thin film devices represent future trends for new technologies. One appealing feature of solution-processed materials is that they can be deposited into thin films using solution-processed deposition techniques that are straightforward, inexpensive, high throughput and advantageous to industrialize thin film based devices. However, solution-processed techniques rely on wet deposition, which has limitations in certain applications, such as multi-layered film deposition of similar materials and blended film deposition of dissimilar materials. These limitations cannot be addressed by traditional, vacuum-based deposition techniques because these dry approaches are often too energetic and can degrade soft materials, such as polymers, such that the performance of resulting thin film based devices is compromised. The work presented in this dissertation explores a novel thin film deposition technique, namely emulsion-based, resonant infrared, matrix-assisted pulsed laser evaporation (RIR-MAPLE), which combines characteristics of wet and dry deposition techniques for solution-processed materials. Previous studies have demonstrated the feasibility of emulsion-based RIR-MAPLE to deposit uniform and continuous organic, nanoparticle and blended films, as well as hetero-structures that otherwise are difficult to achieve. However, fundamental understanding of the growth mechanisms that govern

  6. Films of brookite TiO2 nanorods/nanoparticles deposited by matrix-assisted pulsed laser evaporation as NO2 gas-sensing layers

    Caricato, A. P.; Buonsanti, R.; Catalano, M.; Cesaria, M.; Cozzoli, P. D.; Luches, A.; Manera, M. G.; Martino, M.; Taurino, A.; Rella, R.

    2011-09-01

    Titanium dioxide (TiO2) nanorods in the brookite phase, with average dimensions of 3-4 nm × 20-50 nm, were synthesized by a wet-chemical aminolysis route and used as precursors for thin films that were deposited by the matrix-assisted pulsed laser evaporation (MAPLE) technique. A nanorod solution in toluene (0.016 wt% TiO2) was frozen at the liquid-nitrogen temperature and irradiated with a KrF excimer laser at a fluence of 350 mJ/cm2 and repetition rate of 10 Hz. Single-crystal Si wafers, silica slides, carbon-coated Cu grids and alumina interdigitated slabs were used as substrates to allow performing different characterizations. Films fabricated with 6000 laser pulses had an average thickness of ˜150 nm, and a complete coverage of the selected substrate as achieved. High-resolution scanning and transmission electron microscopy investigations evidenced the formation of quite rough films incorporating individually distinguishable TiO2 nanorods and crystalline spherical nanoparticles with an average diameter of ˜13 nm. Spectrophotometric analysis showed high transparency through the UV-Vis spectral range. Promising resistive sensing responses to 1 ppm of NO2 mixed in dry air were obtained.

  7. Films of brookite TiO{sub 2} nanorods/nanoparticles deposited by matrix-assisted pulsed laser evaporation as NO{sub 2} gas-sensing layers

    Caricato, A.P.; Cesaria, M.; Luches, A.; Martino, M. [University of Salento, Department of Physics, Lecce (Italy); Buonsanti, R. [Istituto di Nanoscienze del CNR, National Nanotechnology Laboratory (NNL), Lecce (Italy); Catalano, M.; Manera, M.G.; Taurino, A.; Rella, R. [IMM-CNR, Institute for Microelectronics and Microsystems, Lecce (Italy); Cozzoli, P.D. [Istituto di Nanoscienze del CNR, National Nanotechnology Laboratory (NNL), Lecce (Italy); University of Salento, Department of Innovation Engineering, Lecce (Italy)

    2011-09-15

    Titanium dioxide (TiO{sub 2}) nanorods in the brookite phase, with average dimensions of 3-4 nm x 20-50 nm, were synthesized by a wet-chemical aminolysis route and used as precursors for thin films that were deposited by the matrix-assisted pulsed laser evaporation (MAPLE) technique. A nanorod solution in toluene (0.016 wt% TiO{sub 2}) was frozen at the liquid-nitrogen temperature and irradiated with a KrF excimer laser at a fluence of 350 mJ/cm{sup 2} and repetition rate of 10 Hz. Single-crystal Si wafers, silica slides, carbon-coated Cu grids and alumina interdigitated slabs were used as substrates to allow performing different characterizations. Films fabricated with 6000 laser pulses had an average thickness of {proportional_to}150 nm, and a complete coverage of the selected substrate as achieved. High-resolution scanning and transmission electron microscopy investigations evidenced the formation of quite rough films incorporating individually distinguishable TiO{sub 2} nanorods and crystalline spherical nanoparticles with an average diameter of {proportional_to}13 nm. Spectrophotometric analysis showed high transparency through the UV-Vis spectral range. Promising resistive sensing responses to 1 ppm of NO{sub 2} mixed in dry air were obtained. (orig.)

  8. TiO2 nanoparticle thin film deposition by matrix assisted pulsed laser evaporation for sensing applications

    Caricato, A.P.; Capone, S.; Ciccarella, G.; Martino, M.; Rella, R.; Romano, F.; Spadavecchia, J.; Taurino, A.; Tunno, T.; Valerini, D.

    2007-01-01

    The MAPLE technique has been used for the deposition of nanostructured titania (TiO 2 ) nanoparticles thin films to be used for gas sensors applications. An aqueous solution of TiO 2 nanoparticles, synthesised by a novel chemical route, was frozen at liquid nitrogen temperature and irradiated with a pulsed ArF excimer laser in a vacuum chamber. A uniform distribution of TiO 2 nanoparticles with an average size of about 10 nm was deposited on Si and interdigitated Al 2 O 3 substrates as demonstrated by high resolution scanning electron microscopy-field emission gun inspection (SEM-FEG). Energy dispersive X-ray (EDX) analysis revealed the presence of only the titanium and oxygen signals and FTIR (Fourier transform infra-red) revealed the TiO 2 characteristic composition and bond. A comparison with a spin coated thin film obtained from the same solution of TiO 2 nanoparticles is reported. The sensing properties of the films deposited on interdigitated substrates were investigated, too

  9. New bio-active, antimicrobial and adherent coatings of nanostructured carbon double-reinforced with silver and silicon by Matrix-Assisted Pulsed Laser Evaporation for medical applications

    Duta, L.; Ristoscu, C.; Stan, G. E.; Husanu, M. A.; Besleaga, C.; Chifiriuc, M. C.; Lazar, V.; Bleotu, C.; Miculescu, F.; Mihailescu, N.; Axente, E.; Badiceanu, M.; Bociaga, D.; Mihailescu, Ion N.

    2018-05-01

    We report on Matrix-Assisted Pulsed Laser Evaporation (MAPLE) deposition of Carbon thin films, simple or reinforced with intended concentrations of Ag and Si. A KrF∗ (λ = 248 nm, τFWHM ≤ 25 ns, ν = 10 Hz) excimer laser was used for irradiation. The effect of a post-deposition thermal treatment in vacuum was studied. Besides detailed morphological, compositional, structural and pull-out adherence characterizations, the potential of the carbonaceous films for medical applications was investigated in vitro by anti-biofilm and cytocompatibility assays. The microscopic images evidenced no delaminations. Micro-Raman spectroscopy revealed a graphitization tendency depending on preparation conditions, thermal treatment and reinforcing agents' presence. Adherence values improved considerably after thermal treatment. In vitro biological evaluation showed that the films containing ∼1.85 at.% Ag were non-cytotoxic for MG63 cells, while eliciting a limited antimicrobial activity. The increase of Ag content to 3.6 at.% results in a significant enhancement of antimicrobial activity, whilst maintaining the cytotoxic action and adherence characteristics at acceptable levels. We propose a new class of metamaterials based on C reinforced with Ag and Si obtained by MAPLE for medical applications, i.e. the prevention and treatment of various infections associated with biofilms developed on implants and other medical equipments.

  10. In vitro behavior of human mesenchymal stem cells on poly(N-isopropylacrylamide) based biointerfaces obtained by matrix assisted pulsed laser evaporation

    Icriverzi, Madalina; Rusen, Laurentiu; Sima, Livia Elena; Moldovan, Antoniu; Brajnicov, Simona; Bonciu, Anca; Mihailescu, Natalia; Dinescu, Maria; Cimpean, Anisoara; Roseanu, Anca; Dinca, Valentina

    2018-05-01

    The use of smart coatings with tunable characteristics in bioengineering fields is directly correlated with the surface chemical and topographical properties, the method of preparation, and also with the type of cells implied for the specific application. In this work, a versatile surface modification technique based on the use of lasers (Matrix-Assisted Pulsed Laser Evaporation (MAPLE)) was used to yield poly(N-isopropylacrylamide) (pNIPAM) and its derivatives (amine, azide and amide terminated pNIPAM) functional and termoresponsive thin films. Surface properties of pNIPAM and its derivative films such as morphology, roughness and hydrophobic/hydrophilic character, as well as the thermoresponsive capacity were investigated by atomic force microscopy and contact angle measurements. The chemical characteristics of the pNIPAM based thin films were analysed by Fourier Transform Infrared Spectroscopy (FTIR). The chemical functionality was kept for all the samples obtained by MAPLE and the thermoresponse was demonstrated by the change in the contact angle and thickness values when the temperature was shifted from 37 °C to 24 °C for all the materials tested, with a smaller change for maleimide terminated pNIPAM. Biological assays performed in vitro (fluorescence microscopy and Scanning Electron Microscopy (SEM)) confirmed the conditioning of the early mesenchymal stem cell (MSC) growth by specific chemistry of the coatings. The cell imaging analysis revealed no cytotoxic effect of pNIPAM surfaces irrespective of type of functionalization. An increased proliferation rate of the cells grown on pNIPAM-azide surfaces and a lower cell density on pNIPAM-maleimide surfaces compared to the pNIPAM surfaces was observed, which can direct their use to potential surfaces in regenerative medicine approaches.

  11. Study of the mechanisms of matrix assisted laser desorption / ionization

    Manuelli, Pascal

    1995-01-01

    This research thesis aims at a better knowledge of some aspects of a complex mechanism: the matrix-assisted laser desorption/ionization (MALDI). The author first proposes a comparative analysis of results obtained by time-of-flight (TOF) mass spectrometry and by Fourier transform mass spectrometry. He reports the study of the matrix role (notably a polymeric matrix) as a matter submitted to laser desorption. In this respect, the influence of the incident wavelength has been studied. The author also reports a comparative of ions produced by matrix laser desorption (study performed by Fourier transform mass spectrometry) and of neutral molecules (study performed by flash pyrolysis coupled with gas chromatography and with mass spectrometry). Finally, results obtained on derivatives and complexes based on beta-cyclodextrins highlight benefits as well as limitations of this technique [fr

  12. Investigations into ultraviolet matrix-assisted laser desorption

    Heise, Theodore W. [Iowa State Univ., Ames, IA (United States)

    1993-07-01

    Matrix-assisted laser desorption (MALD) is a technique for converting large biomolecules into gas phase ions. Some characteristics of the commonly used uv matrices are determined. Solubilities in methanol range from 0.1 to 0.5 M. Solid phase absorption spectra are found to be similar to solution, but slightly red-shifted. Acoustic and quartz crystal microbalance signals are investigated as possible means of uv-MALD quantitation. Evidence for the existence of desorption thresholds is presented. Threshold values are determined to be in the range of 2 to 3 MW/cm2. A transient imaging technique based on laser-excited fluorescence for monitoring MALD plumes is described. Sensitivity is well within the levels required for studying matrix-assisted laser desorption, where analyte concentrations are significantly lower than those in conventional laser desorption. Results showing the effect of film morphology, particularly film thickness, on plume dynamics are presented. In particular, MALD plumes from thicker films tend to exhibit higher axial velocities. Fluorescent labeling of protein and of DNA is used to allow imaging of their uv-MALD generated plumes. Integrated concentrations are available with respect to time, making it possible to assess the rate of fragmentation. The spatial and temporal distributions are important for the design of secondary ionization schemes to enhance ion yields and for the optimization of ion collection in time-of-flight MS instruments to maximize resolution. Such information could also provide insight into whether ionization is closely associated with the desorption step or whether it is a result of subsequent collisions with the matrix gas (e.g., proton transfer). Although the present study involves plumes in a normal atmosphere, adaptation to measurements in vacuum (e.g., inside a mass spectrometer) should be straightforward.

  13. Is energy pooling necessary in ultraviolet matrix-assisted laser desorption/ionization?

    Lin, Hou-Yu; Song, Botao; Lu, I-Chung; Hsu, Kuo-Tung; Liao, Chih-Yu; Lee, Yin-Yu; Tseng, Chien-Ming; Lee, Yuan-Tseh; Ni, Chi-Kung

    2014-01-15

    Energy pooling has been suggested as the key process for generating the primary ions during ultraviolet matrix-assisted laser desorption/ionization (UV-MALDI). In previous studies, decreases in fluorescence quantum yields as laser fluence increased for 2-aminobenzoic acid, 2,5-dihydroxybenzoic acid (2,5-DHB), and 3-hydroxypicolinic acid were used as evidence of energy pooling. This work extends the research to other matrices and addresses whether energy pooling is a universal property in UV-MALDI. Energy pooling was investigated in a time-resolved fluorescence experiment by using a short laser pulse (355 nm, 20 ps pulse width) for excitation and a streak camera (1 ps time resolution) for fluorescence detection. The excited-state lifetime of 2,5-DHB decreased with increases in laser fluence. This suggests that a reaction occurs between two excited molecules, and that energy pooling may be one of the possible reactions. However, the excited-state lifetime of 2,4,6-trihydroxyacetophenone (THAP) did not change with increases in laser fluence. The upper limit of the energy pooling rate constant for THAP is estimated to be approximately 100-500 times smaller than that of 2,5-DHB. The small energy pooling rate constant for THAP indicates that the potential contribution of the energy pooling mechanism to the generation of THAP matrix primary ions should be reconsidered. Copyright © 2013 John Wiley & Sons, Ltd.

  14. Ejection of matrix-polymer clusters in matrix-assisted laser evaporation: Experimental observations

    Sellinger, Aaron T; Leveugle, Elodie; Gogick, Kristy; Peman, Guillaume; Zhigilei, Leonid V; Fitz-Gerald, James M

    2007-01-01

    The morphology of polymer films deposited with the matrix-assisted pulsed laser evaporation (MAPLE) technique is explored for various target compositions and laser fluences. Composite targets of 1 to 5 wt.% poly(methyl methacrylate), PMMA, dissolved in a volatile matrix material, toluene, were ablated using an excimer laser at fluences ranging from 0.045 J/cm 2 to 0.75 J/cm 2 . Films were deposited on Si substrates at room temperature in a dynamic 100 mTorr Ar atmosphere. Scanning electron microscopy (SEM) imaging revealed that the morphology of the deposited films varied significantly with both laser fluence and PMMA concentration. The morphologies of large deposited particles were similar to that of deflated ''balloons''. It is speculated that during ablation of the frozen target, clusters comprised of both polymer and solvent ranging from 100 nm to 10 μm in size are ejected and deposited onto the substrate. The solvent begins to evaporate from the clusters during flight from the target, but does not completely evaporate until deposited on the room temperature substrate. The dynamics of the toluene evaporation may lead to the formation of the deflated structures. This explanation is supported by the observation of stable polymer-matrix droplets ejected in molecular dynamics simulations of MAPLE

  15. Matrix-assisted laser desorption fourier transform mass spectrometry for biological compounds

    Hettich, R.; Buchanan, M.

    1990-01-01

    The recent development of matrix-assisted UV laser desorption (LD) mass spectrometry has made possible the ionization and detection of extremely large molecules (with molecular weights exceeding 100,000 Daltons). This technique has generated enormous interest in the biological community for the direct examination of large peptides and oligonucleotides. Although this matrix-assisted ionization method has been developed and used almost exclusively with time-of-flight (TOF) mass spectrometers, research is currently in progress to demonstrate this technique with trapped ion mass spectrometers, such as Fourier transform ion cyclotron resonance mass spectrometry (FTMS). The potential capabilities of FTMS for wide mass range, high resolution measurement, and ion trapping experiments suggest that this instrumental technique should be useful for the detailed structural characterization of large ions generated by the matrix-assisted technique. We have recently demonstrated that matrix-assisted ultraviolet laser desorption can be successfully used with FTMS for the ionization of small peptides. The objective of this report is to summarize the application and current limitations of matrix-assisted laser desorption FTMS for the characterization of peptides and oligonucleotides at the isomeric level. 4 refs., 3 figs., 2 tabs.

  16. Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry

    Baar, B.L.M. van

    2000-01-01

    Chemical analysis for the characterisation of micro-organisms is rapidly evolving, after the recent advent of new ionisation methods in mass spectrometry (MS): electrospray (ES) and matrix-assisted laser desorption/ionisation (MALDI). These methods allow quick characterisation of micro-organisms,

  17. Multisample matrix-assisted laser desorption source for molecular beams of neutral peptides

    Lupulescu, C.; Abd El Rahim, M.; Antoine, R.; Barbaire, M.; Broyer, M.; Dagany, X.; Maurelli, J.; Rayane, D.; Dugourd, Ph.

    2006-01-01

    We developed and tested a multisample laser desorption source for producing stable molecular beams of neutral peptides. Our apparatus is based on matrix-assisted laser desorption technique. The source consists of 96 different targets which may be scanned by a software control procedure. Examples of molecular beams of neutral peptides are presented, as well as the influence of the different source parameters on the jet

  18. Matrix-assisted laser-desorption-ionization mass spectrometry of proteins using a free-electron laser

    Cramer, R.; Hillenkamp, F.; Haglund, R.

    1995-01-01

    Matrix-assisted laser desorption-ionization (MALDI) mass spectrometry (MS) is one of the most promising techniques for spectral fingerprinting large molecules, such as proteins, oligonucleotides and carbohydrates. In the usual implementation of this technique, the analyte molecule is dissolved in an aromatic liquid matrix material which resonantly absorbs ultraviolet laser light. Resonant absorption by π-π* transitions volatilizes the matrix and initiates subsequent charge transfer to the analyte molecules, which are detected by time-of-flight mass spectrometry. Recent MALDI-MS studies with Er:YAG (2.94 μm) and CO 2 4 (9.4-10.6 μm) lasers suggest that them is significant unexplored potential for mass spectrometry of macromolecules, including oligonucleotide, in the mid-infrared. Preliminary experiments show that it is possible to capitalize on the rich rovibronic absorption spectrum of virtually all organics to initiate resonant desorption in matrix material over the entire range of pH values. However, the mechanism of charge transfer is particularly problematic for infrared MALDI because of the low photon energy. In this paper, we report the results of MALI-MS studies on small proteins using the Vanderbilt FEL and several matrix materials. Proteins with masses up to roughly 6,000 amu were detected with high resolution in a linear time-of-flight mass spectrometer. By varying the pulse duration using a broadband Pockels cell, we have been able to compare the results of relatively long (5 μs) and short (0.1 μs) irradiation on the desorption and ionization processes. Compared to uv-MALDI spectra of identical analytes obtained with a nitrogen laser (337 nm) in the same time-of-flight spectrometer, the infrared results appear to show that the desorption and ionization process goes on over a somewhat longer time scale

  19. Ghost peaks observed after atmospheric pressure matrix-assisted laser desorption/ionization experiments may disclose new ionization mechanism of matrix-assisted hypersonic velocity impact ionization.

    Moskovets, Eugene

    2015-08-30

    Understanding the mechanisms of matrix-assisted laser desorption/ionization (MALDI) promises improvements in the sensitivity and specificity of many established applications in the field of mass spectrometry. This paper reports a serendipitous observation of a significant ion yield in a post-ionization experiment conducted after the sample had been removed from a standard atmospheric pressure (AP)-MALDI source. This post-ionization is interpreted in terms of collisions of microparticles moving with a hypersonic velocity into a solid surface. Calculations show that the thermal energy released during such collisions is close to that absorbed by the top matrix layer in traditional MALDI. The microparticles, containing both the matrix and analytes, could be detached from a film produced inside the inlet capillary during the sample ablation and accelerated by the flow rushing through the capillary. These observations contribute some new perspective to ion formation in both laser and laser-less matrix-assisted ionization. An AP-MALDI ion source hyphenated with a three-stage high-pressure ion funnel system was utilized for peptide mass analysis. After the laser had been turned off and the MALDI sample removed, ions were detected during a gradual reduction of the background pressure in the first funnel. The constant-rate pressure reduction led to the reproducible appearance of different singly and doubly charged peptide peaks in mass spectra taken a few seconds after the end of the MALDI analysis of a dried-droplet spot. The ion yield as well as the mass range of ions observed with a significant delay after a completion of the primary MALDI analysis depended primarily on the background pressure inside the first funnel. The production of ions in this post-ionization step was exclusively observed during the pressure drop. A lower matrix background and significant increase in relative yield of double-protonated ions are reported. The observations were partially consistent

  20. Electrospray ionization and matrix assisted laser desorption/ionization mass spectrometry: powerful analytical tools in recombinant protein chemistry

    Andersen, Jens S.; Svensson, B; Roepstorff, P

    1996-01-01

    Electrospray ionization and matrix assisted laser desorption/ionization are effective ionization methods for mass spectrometry of biomolecules. Here we describe the capabilities of these methods for peptide and protein characterization in biotechnology. An integrated analytical strategy is presen......Electrospray ionization and matrix assisted laser desorption/ionization are effective ionization methods for mass spectrometry of biomolecules. Here we describe the capabilities of these methods for peptide and protein characterization in biotechnology. An integrated analytical strategy...... is presented encompassing protein characterization prior to and after cloning of the corresponding gene....

  1. A Simple Sonication Improves Protein Signal in Matrix-Assisted Laser Desorption Ionization Imaging

    Lin, Li-En; Su, Pin-Rui; Wu, Hsin-Yi; Hsu, Cheng-Chih

    2018-02-01

    Proper matrix application is crucial in obtaining high quality matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI). Solvent-free sublimation was essentially introduced as an approach of homogeneous coating that gives small crystal size of the organic matrix. However, sublimation has lower extraction efficiency of analytes. Here, we present that a simple sonication step after the hydration in standard sublimation protocol significantly enhances the sensitivity of MALDI MSI. This modified procedure uses a common laboratory ultrasonicator to immobilize the analytes from tissue sections without noticeable delocalization. Improved imaging quality with additional peaks above 10 kDa in the spectra was thus obtained upon sonication treatment. [Figure not available: see fulltext.

  2. Molecular dynamics simulations of matrix assisted laser desorption ionization: Matrix-analyte interactions

    Nangia, Shivangi; Garrison, Barbara J.

    2011-01-01

    There is synergy between matrix assisted laser desorption ionization (MALDI) experiments and molecular dynamics (MD) simulations. To understand analyte ejection from the matrix, MD simulations have been employed. Prior calculations show that the ejected analyte molecules remain solvated by the matrix molecules in the ablated plume. In contrast, the experimental data show free analyte ions. The main idea of this work is that analyte molecule ejection may depend on the microscopic details of analyte interaction with the matrix. Intermolecular matrix-analyte interactions have been studied by focusing on 2,5-dihydroxybenzoic acid (DHB; matrix) and amino acids (AA; analyte) using Chemistry at HARvard Molecular Mechanics (CHARMM) force field. A series of AA molecules have been studied to analyze the DHB-AA interaction. A relative scale of AA molecule affinity towards DHB has been developed.

  3. Direct Surface Analysis of Fungal Species by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

    Valentine, Nancy B.(BATTELLE (PACIFIC NW LAB)); Wahl, Jon H.(BATTELLE (PACIFIC NW LAB)); Kingsley, Mark T.(BATTELLE (PACIFIC NW LAB)); Wahl, Karen L.(BATTELLE (PACIFIC NW LAB))

    2001-12-01

    Intact spores and/or hyphae of Aspergillus niger, Rhizopus oryzae, Trichoderma reesei and Phanerochaete chrysosporium are analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). This study investigates various methods of sample preparation and matrices to determine optimum collection and analysis criteria for fungal analysis by MALDI-MS. Fungi are applied to the MALDI sample target as untreated, sonicated, acid/heat treated, or blotted directly from the fungal culture with double-stick tape. Ferulic acid or sinapinic acid matrix solution is layered over the dried samples and analyzed by MALDI-MS. Statistical analysis of the data show that simply using double stick tape to collect and transfer to a MALDI sample plate typically worked as well as the other preparation methods, but requires the least sample handling.

  4. MALDI (matrix assisted laser desorption ionization) Imaging Mass Spectrometry (IMS) of skin: Aspects of sample preparation.

    de Macedo, Cristiana Santos; Anderson, David M; Schey, Kevin L

    2017-11-01

    MALDI (matrix assisted laser desorption ionization) Imaging Mass Spectrometry (IMS) allows molecular analysis of biological materials making possible the identification and localization of molecules in tissues, and has been applied to address many questions on skin pathophysiology, as well as on studies about drug absorption and metabolism. Sample preparation for MALDI IMS is the most important part of the workflow, comprising specimen collection and preservation, tissue embedding, cryosectioning, washing, and matrix application. These steps must be carefully optimized for specific analytes of interest (lipids, proteins, drugs, etc.), representing a challenge for skin analysis. In this review, critical parameters for MALDI IMS sample preparation of skin samples will be described. In addition, specific applications of MALDI IMS of skin samples will be presented including wound healing, neoplasia, and infection. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. A study on matrix assisted pulsed evaporation (MAPLE) of organic materials

    Matei, Andreea; Canulescu, Stela; Constantinescu, Catalin

    2012-01-01

    Organic films can be produced either by MAPLE or directly by PLD (Pulsed laser deposition). For a reasonable deposition rate of ng/cm2 per pulse for film production by MAPLE a fluence of 1-1.5 J/cm2 is required at the laser wavelength of 355 nm, while the fluence can be considerably lower at 248 nm....... At high fluence the deposition rate of proteins by MAPLE seems to decrease. The surface roughness is still an issue, but at low fluence it seems to be acceptable. The fragmentation rate increases with fluence, and seems to be less pronounced for MAPLE than for PLD. Also this issue is not yet resolved....

  6. Generation of CsI cluster ions for mass calibration in matrix-assisted laser desorption/ionization mass spectrometry

    Lou, X.; Dongen, van J.L.J.; Meijer, E.W.

    2010-01-01

    A simple method was developed for the generation of cesium iodide (CsI) cluster ions up to m/z over 20,000 in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Calibration ions in both positive and negative ion modes can readily be generated from a single MALDI spot of CsI(3)

  7. Development of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) for plant metabolite analysis

    Korte, Andrew R [Iowa State Univ., Ames, IA (United States)

    2014-12-01

    This thesis presents efforts to improve the methodology of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) as a method for analysis of metabolites from plant tissue samples. The first chapter consists of a general introduction to the technique of MALDI-MSI, and the sixth and final chapter provides a brief summary and an outlook on future work.

  8. Phenotypic identification of Porphyromonas gingivalis validated with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Rams, Thomas E; Sautter, Jacqueline D; Getreu, Adam; van Winkelhoff, Arie J

    OBJECTIVE: Porphyromonas gingivalis is a major bacterial pathogen in human periodontitis. This study used matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to assess the accuracy of a rapid phenotypic identification scheme for detection of cultivable P.

  9. Calibration of matrix-assisted laser desorption/ionization time-of-flight peptide mass fingerprinting spectra

    Hjernø, Karin; Højrup, Peter

    2007-01-01

    This chapter describes a number of aspects important for calibration of matrix-assisted laser desorption/ionization time-of-flight spectra prior to peptide mass fingerprinting searches. Both multipoint internal calibration and mass defect-based calibration is illustrated. The chapter describes ho...

  10. Characterization of polyesters by matrix-assisted laser desorption/ionization and Fourier transform mass spectrometry.

    Mize, Todd H; Simonsick, William J; Amster, I Jonathan

    2003-01-01

    Two homopolyesters, poly(neopentyl glycol-alt-isophthalic acid) and poly(hexanediol-alt-azelaic acid), and two copolyesters, poly(dipropoxylated bisphenol-A-alt-(isophthalic acid-co-adipic acid)) and poly(neopentyl glycol-alt-(adipic acid-co-isophthalic acid)) were analyzed by internal source matrix assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS). The high resolution and high mass accuracy provided by FTMS greatly facilitate the characterization of the polyester and copolyester samples. Isobaric resolution allows the ion abundances of overlapping isotopic envelopes to be assessed. Repeat units were confirmed and end functionality assigned. Single shot mass spectra of the entire polymeric distribution demonstrate that the dynamic range of this internal MALDI source instrument and the analyzer cell exceeds performance of those previously reported for higher field instruments. Corrections of space charge mass shift effects are demonstrated for the analytes using an external calibrant and (subsequent to confirmation of structure) via internal calibration which removes ambiguity due to space charge differences in calibrant and analyte spectra. Capillary gel permeation chromatography was used to prepare low polydispersity samples from a high polydispersity polyester, improving the measurement of molecular weight distribution two-fold while retaining the benefits of high resolution mass spectrometry for elucidation of oligomer identity.

  11. Study of ionization process of matrix molecules in matrix-assisted laser desorption ionization

    Murakami, Kazumasa; Sato, Asami; Hashimoto, Kenro; Fujino, Tatsuya, E-mail: fujino@tmu.ac.jp

    2013-06-20

    Highlights: ► Proton transfer and adduction reaction of matrix in MALDI were studied. ► Hydroxyl group forming intramolecular hydrogen bond was related to the ionization. ► Intramolecular proton transfer in the electronic excited state was the initial step. ► Non-volatile analytes stabilized protonated matrix in the ground state. ► A possible mechanism, “analyte support mechanism”, has been proposed. - Abstract: Proton transfer and adduction reaction of matrix molecules in matrix-assisted laser desorption ionization were studied. By using 2,4,6-trihydroxyacetophenone (THAP), 2,5-dihydroxybenzoic acid (DHBA), and their related compounds in which the position of a hydroxyl group is different, it was clarified that a hydroxyl group forming an intramolecular hydrogen bond is related to the ionization of matrix molecules. Intramolecular proton transfer in the electronic excited state of the matrix and subsequent proton adduction from a surrounding solvent to the charge-separated matrix are the initial steps for the ionization of matrix molecules. Nanosecond pump–probe NIR–UV mass spectrometry confirmed that the existence of analyte molecules having large dipole moment in their structures is necessary for the stabilization of [matrix + H]{sup +} in the electronic ground state.

  12. Focused Electrospray Deposition for Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

    Jeong, Kyung Hwan; Seo, Jong Cheol; Yoon, Hye Joo; Shin, Seung Koo

    2010-01-01

    Focused electrospray (FES) deposition method is presented for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. FES ion optics consists of two cylindrical focusing electrodes capped with a truncated conical electrode through which an electrospray emitter passes along the cylindrical axis. A spray of charged droplets is focused onto a sample well on a MALDI target plate under atmospheric pressure. The shape and size distributions of matrix crystals are visualized by scanning electron microscope and the mass spectra are obtained by time-of-flight mass spectrometry. Angiotensin II, bradykinin, and substance P are used as test samples, while α-cyano-4-hydroxycinnamic acid and dihydroxybenzoic acid are employed as matrices. FES of a sample/matrix mixture produces fine crystal grains on a 1.3 mm spot and reproducibly yields the mass spectra with little shot-to-shot and spot-to-spot variations. Although FES greatly stabilizes the signals, the space charge due to matrix ions limits the detection sensitivity of peptides. To avoid the space charge problem, we adopted a dual FES/FES mode, which separately deposits matrix and sample by FES in sequence. The dual FES/FES mode reaches the detection sensitivity of 0.88 amol, enabling ultrasensitive detection of peptides by homogeneously depositing matrix and sample under atmospheric pressure

  13. Matrix-assisted laser desorption/ionization mass spectrometry for the structural characterization of modified oligonucleotides

    Hurst, G.B.; Hettich, R.L.; Buchanan, M.V.; Stemmler, E.A.

    1993-01-01

    Matrix-assisted laser desorption ionization (MALDI) Fourier transform ion cyclotron resonance mass spectrometry (FTMS) and MALDI time-of-flight mass spectrometry (TOFMS) are being used to characterize conditions for the efficient desorption and ionization of normal and modified nucleic acid components. Basic and acidic matrix materials have been evaluated on the components. Basic and acidic matrix materials have been evaluated on the FTMS and TOFMS. Using MALDI-FTMS at 355 nm, less fragmentation has been observed using 2,5-dihydroxybenzoic acid, while more extensive fragmentation is observed for basic matrices, such as 1,5-diaminonaphthalene and 9-aminophenanthrene. Elevation of the cell pressure by the addition of Ar or CO 2 provides collisional cooling of desorbed ions, resulting in an enhancement of [M--H] - and structurally significant high-mass fragment ions. Using MALDI-TOFMS at 337 nm, fragmentation is significantly reduced relative to that observed on the FTMS, perhaps as a consequence of the longer times required for FTMS detection. On the FTMS and TOFMS, cluster ions have been observed in the negative ion mode when metal ions are present in the 2,5-dihydroxybenzoic acid matrix. Metal ion additions and clusters with matrix salts have also been observed for dinucleotides. Applications of MALDI-FTMS and MALDI-TOF to the detection of hydroxylated PAH nucleoside adducts are presented

  14. Drawing a different picture with pencil lead as matrix-assisted laser desorption/ionization matrix for fullerene derivatives.

    Nye, Leanne C; Hungerbühler, Hartmut; Drewello, Thomas

    2018-02-01

    Inspired by reports on the use of pencil lead as a matrix-assisted laser desorption/ionization matrix, paving the way towards matrix-free matrix-assisted laser desorption/ionization, the present investigation evaluates its usage with organic fullerene derivatives. Currently, this class of compounds is best analysed using the electron transfer matrix trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene] malononitrile (DCTB), which was employed as the standard here. The suitability of pencil lead was additionally compared to direct (i.e. no matrix) laser desorption/ionization-mass spectrometry. The use of (DCTB) was identified as the by far gentler method, producing spectra with abundant molecular ion signals and much reduced fragmentation. Analytically, pencil lead was found to be ineffective as a matrix, however, appears to be an extremely easy and inexpensive method for producing sodium and potassium adducts.

  15. Detection of Bacteriocins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

    Rose, Natisha L.; Sporns, Peter; McMullen, Lynn M.

    1999-01-01

    The use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the detection of bacteriocins was investigated. A 30-s water wash of the sample on the MALDI-TOF MS probe was effective in removing contaminants of the analyte. This method was used for rapid detection of nisin, pediocin, brochocin A and B, and enterocin A and B from culture supernatants and for detection of enterocin B throughout its purification.

  16. [Special application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry in clinical microbiological diagnostics].

    Nagy, Erzsébet; Abrók, Marianna; Bartha, Noémi; Bereczki, László; Juhász, Emese; Kardos, Gábor; Kristóf, Katalin; Miszti, Cecilia; Urbán, Edit

    2014-09-21

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry as a new possibility for rapid identification of bacteria and fungi revolutionized the clinical microbiological diagnostics. It has an extreme importance in the routine microbiological laboratories, as identification of the pathogenic species rapidly will influence antibiotic selection before the final determination of antibiotic resistance of the isolate. The classical methods for identification of bacteria or fungi, based on biochemical tests, are influenced by many environmental factors. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a rapid method which is able to identify a great variety of the isolated bacteria and fungi based on the composition of conserved ribosomal proteins. Recently several other applications of the method have also been investigated such as direct identification of pathogens from the positive blood cultures. There are possibilities to identify bacteria from the urine samples in urinary tract infection or from other sterile body fluids. Using selective enrichment broth Salmonella sp from the stool samples can be identified more rapidly, too. The extended spectrum beta-lactamase or carbapenemase production of the isolated bacteria can be also detected by this method helping the antibiotic selection in some cases. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry based methods are suitable to investigate changes in deoxyribonucleic acid or ribonucleic acid, to carry out rapid antibiotic resistance determination or other proteomic analysis. The aim of this paper is to give an overview about present possibilities of using this technique in the clinical microbiological routine procedures.

  17. Current status of matrix-assisted laser desorption ionisation-time of flight mass spectrometry in the clinical microbiology laboratory.

    Kok, Jen; Chen, Sharon C A; Dwyer, Dominic E; Iredell, Jonathan R

    2013-01-01

    The integration of matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) into many clinical microbiology laboratories has revolutionised routine pathogen identification. MALDI-TOF MS complements and has good potential to replace existing phenotypic identification methods. Results are available in a more clinically relevant timeframe, particularly in bacteraemic septic shock. Novel applications include strain typing and the detection of antimicrobial resistance, but these are not widely used. This review discusses the technical aspects, current applications, and limitations of MALDI-TOF MS.

  18. Matrix-assisted laser desorption/ionisation mass spectrometry imaging and its development for plant protein imaging

    Millar A Harvey

    2011-07-01

    Full Text Available Abstract Matrix-Assisted Laser Desorption/Ionisation (MALDI mass spectrometry imaging (MSI uses the power of high mass resolution time of flight (ToF mass spectrometry coupled to the raster of lasers shots across the cut surface of tissues to provide new insights into the spatial distribution of biomolecules within biological tissues. The history of this technique in animals and plants is considered and the potential for analysis of proteins by this technique in plants is discussed. Protein biomarker identification from MALDI-MSI is a challenge and a number of different approaches to address this bottleneck are discussed. The technical considerations needed for MALDI-MSI are reviewed and these are presented alongside examples from our own work and a protocol for MALDI-MSI of proteins in plant samples.

  19. Rapid detection of undesired cosmetic ingredients by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Ouyang, Jie; An, Dongli; Chen, Tengteng; Lin, Zhiwei

    2017-10-01

    In recent years, cosmetic industry profits soared due to the widespread use of cosmetics, which resulted in illicit manufacturers and products of poor quality. Therefore, the rapid and accurate detection of the composition of cosmetics has become crucial. At present, numerous methods, such as gas chromatography and liquid chromatography-mass spectrometry, were available for the analysis of cosmetic ingredients. However, these methods present several limitations, such as failure to perform comprehensive and rapid analysis of the samples. Compared with other techniques, matrix-assisted laser desorption ionization time-of-flight mass spectrometry offered the advantages of wide detection range, fast speed and high accuracy. In this article, we briefly summarized how to select a suitable matrix and adjust the appropriate laser energy. We also discussed the rapid identification of undesired ingredients, focusing on antibiotics and hormones in cosmetics.

  20. Benefits of 2.94 μm infrared matrix-assisted laser desorption/ionization for analysis of labile molecules by Fourier transform mass spectrometry

    Budnik, Bogdan A.; Jensen, Kenneth Bendix; Jørgensen, Thomas J. D.

    2000-01-01

    A 2.94 microm Er:YAG laser was used together with a commercial Fourier transform mass spectrometer to study labile biomolecules. The combination has shown superior performance over conventional 337 nm ultraviolet matrix-assisted laser desorption/ionization (UV-MALDI) Fourier transform mass...

  1. Quantitation of peptides and proteins by matrix-assisted laser desorption/ionization mass spectrometry using (18)O-labeled internal standards

    Mirgorodskaya, O A; Kozmin, Y P; Titov, M I

    2000-01-01

    A method for quantitating proteins and peptides in the low picomole and sub-picomole range has been developed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) with internal (18)O-labeled standards. A simple procedure is proposed to produce such internal standards for...... inhibitor, were quantified by MALDI-time-of-flight (TOF) mass spectrometry.......A method for quantitating proteins and peptides in the low picomole and sub-picomole range has been developed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) with internal (18)O-labeled standards. A simple procedure is proposed to produce such internal standards...

  2. On plate graphite supported sample processing for simultaneous lipid and protein identification by matrix assisted laser desorption ionization mass spectrometry.

    Calvano, Cosima Damiana; van der Werf, Inez Dorothé; Sabbatini, Luigia; Palmisano, Francesco

    2015-05-01

    The simultaneous identification of lipids and proteins by matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS) after direct on-plate processing of micro-samples supported on colloidal graphite is demonstrated. Taking advantages of large surface area and thermal conductivity, graphite provided an ideal substrate for on-plate proteolysis and lipid extraction. Indeed proteins could be efficiently digested on-plate within 15 min, providing sequence coverages comparable to those obtained by conventional in-solution overnight digestion. Interestingly, detection of hydrophilic phosphorylated peptides could be easily achieved without any further enrichment step. Furthermore, lipids could be simultaneously extracted/identified without any additional treatment/processing step as demonstrated for model complex samples such as milk and egg. The present approach is simple, efficient, of large applicability and offers great promise for protein and lipid identification in very small samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Performance of matrix-assisted laser desorption-time of flight mass spectrometry for identification of clinical yeast isolates

    Rosenvinge, Flemming S; Dzajic, Esad; Knudsen, Elisa

    2013-01-01

    Accurate and fast yeast identification is important when treating patients with invasive fungal disease as susceptibility to antifungal agents is highly species related. Matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF-MS) provides a powerful tool with a clear potential...... spectra output, all 13 isolates were correctly identified, resulting in an overall identification performance of 92%. No misidentifications occurred with the two systems. Of the routine isolates one laboratory identified 99/99 (100%) and 90/99 (91%) to species level by Saramis/Axima and conventional...... identification, respectively, whereas the other laboratory identified 83/98 (85%) to species level by both BioTyper/Bruker and conventional identification. Both MALDI-TOF-MS systems are fast, have built-in databases that cover the majority of clinically relevant Candida species, and have an accuracy...

  4. Pigments and proteins in green bacterial chlorosomes studied by matrix-assisted laser desorption ionization mass spectrometry

    Persson, S; Sönksen, C P; Frigaard, N U

    2000-01-01

    We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for mass determination of pigments and proteins in chlorosomes, the light-harvesting organelles from the photosynthetic green sulfur bacterium Chlorobium tepidum. By applying a small volume (1...... microL) of a concentrated suspension of isolated chlorosomes directly to the target of the mass spectrometer we have been able to detect bacteriochlorophyll a and all the major homologs of bacteriochlorophyll c. The peak heights of the different bacteriochlorophyll c homologs in the MALDI spectra were...... proportional to peak areas obtained from HPLC analysis of the same sample. The same result was also obtained when whole cells of Chl. tepidum were applied to the target, indicating that MALDI-MS can provide a rapid method for obtaining a semiquantitative determination or finger-print of the bacteriochlorophyll...

  5. Structural characterization of phospholipids by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry.

    Marto, J A; White, F M; Seldomridge, S; Marshall, A G

    1995-11-01

    Matrix-assisted laser desorption/ionization (MALDI) Fourier transform ion cyclotron resonance mass spectrometry provides for structural analysis of the principal biological phospholipids: glycerophosphatidylcholine, -ethanolamine, -serine, and -inositol. Both positive and negative molecular or quasimolecular ions are generated in high abundance. Isolated molecular ions may be collisionally activated in the source side of a dual trap mass analyzer, yielding fragments serving to identify the polar head group (positive ion mode) and fatty acid side chains (negative ion mode). Azimuthal quadrupolar excitation following collisionally activated dissociation refocuses productions close to the solenoid axis; subsequent transfer of product ions to the analyzer ion trap allows for high-resolution mass analysis. Cyro-cooling of the sample probe with liquid nitrogen greatly reduces matrix adduction encountered in the negative ion mode.

  6. Matrix-Assisted Laser Desorption/Ionization Imaging Mass Spectrometry for the Investigation of Proteins and Peptides

    Burnum, Kristin E.; Frappier, Sara L.; Caprioli, Richard M.

    2008-07-01

    Mass spectrometry (MS) is an excellent technology for molecular imaging because of its high data dimensionality. MS can monitor thousands of individual molecular data channels measured as mass-to-charge (m/z). We describe the use of matrix-assisted laser desorption/ionization (MALDI) MS for the image analysis of proteins, peptides, lipids, drugs, and metabolites in tissues. We discuss the basic instrumentation and sample preparation methods needed to produce high-resolution images and high image reproducibility. Matrix-addition protocols are briefly discussed along with normal operating procedures, and selected biological and medical applications of MALDI imaging MS are described. We give examples of both two- and three-dimensional imaging, including normal mouse embryo implantation, sperm maturation in mouse epididymis, protein distributions in brain sections, protein alterations as a result of drug administration, and protein changes in brain due to neurodegeneration and tumor formation. Advantages of this technology and future challenges for its improvement are discussed.

  7. Identification of Fatty Acids, Phospholipids, and Their Oxidation Products Using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry and Electrospray Ionization Mass Spectrometry

    Harmon, Christopher W.; Mang, Stephen A.; Greaves, John; Finlayson-Pitts, Barbara J.

    2010-01-01

    Electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) have found increasing application in the analysis of biological samples. Using these techniques to solve problems in analytical chemistry should be an essential component of the training of undergraduate chemists. We…

  8. Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging for Peptide and Protein Analyses: A Critical Review of On-Tissue Digestion

    Cillero-Pastor, B.; Heeren, R.M.A.

    2013-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has established itself among the plethora of mass spectrometry applications. In the biomedical field, MALDI-MSI is being more frequently recognized as a new method for the discovery of biomarkers and targets of

  9. Analysis of Phospholipid Mixtures from Biological Tissues by Matrix-Assisted Laser Desorption and Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS): A Laboratory Experiment

    Eibisch, Mandy; Fuchs, Beate; Schiller, Jurgen; Sub, Rosmarie; Teuber, Kristin

    2011-01-01

    Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used to investigate the phospholipid (PL) compositions of tissues and body fluids, often without previous separation of the total mixture into the individual PL classes. Therefore, the questions of whether all PL classes are detectable…

  10. Determination of saccharides in fruit juices by capillary electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry

    Žídková, Jitka; Chmelík, Josef

    2001-01-01

    Roč. 36, č. 4 (2001), s. 417-421 ISSN 1076-5174 R&D Projects: GA MZe EP9410 Institutional research plan: CEZ:AV0Z4031919 Keywords : fruit juices * adulteration * matrix-assisted laser deasorption Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 2.685, year: 2001

  11. Differentiation of Clinically Relevant mucorales Rhizopus microsporus and R. arrhizus by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)

    Dolatabadi, S.; Kolecka, A.; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

    This study addresses the usefulness of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) for reliable identification of the two most frequently occuring clinical species of Rhizopus, namely R. arrhizus with its two varieties arrhizus and delemar and R.

  12. Characterization of O-glycosylated precursors of insulin-like growth factor II by matrix-assisted laser desorption/ionization mass spectrometry

    Jespersen, S.; Koedam, J.A.; Hoogerbrugge, C.M.; Tjaden, U.R.; Greef, J. van der; Brande, J.L. van den

    1996-01-01

    High molecular weight precursors of insulin-like growth factor II (IGF-II) were isolated from Cohn fraction IV of human plasma by ultrafiltration, affinity chromatography and reversed-phase high-performance liquid chromatography. Molecular weight determination by matrix-assisted laser

  13. Alkaloid profiling of the Chinese herbal medicine Fuzi by combination of matrix-assisted laser desorption ionization mass spectrometry with liquid chromatography-mass spectrometry

    Wang, J.; Heijden, R. van der; Spijksma, G.; Reijmers, T.; Wang, M.; Xu, G.; Hankemeier, T.; Greef, J. van der

    2009-01-01

    A matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) method was developed for the high throughput and robust qualitative profiling of alkaloids in Fuzi-the processed lateral roots of the Chinese herbal medicine Aconitum carmichaeli Debx (A. carmichaeli). After optimization,

  14. Direct analysis of triterpenes from high-salt fermented cucumbers using infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI)

    High-salt samples present a challenge to mass spectrometry (MS) analysis, particularly when electrospray ionization (ESI) is used, requiring extensive sample preparation steps such as desalting, extraction, and purification. In this study, infrared matrix-assisted laser desorption electrospray ioniz...

  15. Identification of Wheat Varieties Using Matrix-assisted Laser Desorption/Ionisation Time-of-flight Mass Spectrometry and an Artificial Neural network

    Bloch, Helle Aagaard; Kesmir, Can; Petersen, Marianne Kjerstine

    1999-01-01

    A novel tool for variety identification of wheat (Triticum aestivum L,) has been developed: an artificial neural network (ANN) is used to classify the gliadin fraction analysed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). The robustness...

  16. DIFFERENTIATION OF AEROMONAS ISOLATES OBTAINED FROM DRINKING WATER DISTRIBUTION SYSTEM USING MATRIX-ASSISTED LASER DESCRIPTION/IONIZATION-MASS SPECTROMETRY (MALDI-MS)

    The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the...

  17. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry identification of large colony beta-hemolytic streptococci containing Lancefield groups A, C, and G

    Salgård Jensen, Christian; Dam-Nielsen, Casper; Arpi, Magnus

    2015-01-01

    BACKGROUND: The aim of this study was to investigate whether large colony beta-hemolytic streptococci containing Lancefield groups A, C, and G can be adequately identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-ToF). Previous studies show varying...

  18. The Characterization of Laser Ablation Patterns and a New Definition of Resolution in Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS).

    O'Rourke, Matthew B; Raymond, Benjamin B A; Padula, Matthew P

    2017-05-01

    Matrix assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) is a technique that has seen a sharp rise in both use and development. Despite this rapid adoption, there have been few thorough investigations into the actual physical mechanisms that underlie the acquisition of IMS images. We therefore set out to characterize the effect of IMS laser ablation patterns on the surface of a sample. We also concluded that the governing factors that control spatial resolution have not been correctly defined and therefore propose a new definition of resolution. Graphical Abstract ᅟ.

  19. Bioaerosol detection by aerosol TOF-mass spectrometry: Application of matrix assisted laser desorption/ionisation

    Wuijckhuijse, A.L. van; Stowers, M.A.; Kientz, Ch.E.; Marijnissen, J.C.M.; Scarlett, B.

    2000-01-01

    In previous publications the use of an aerosol time of flight mass spectrometer was reported for the on-line measurements of aerosols (Weiss 1997, Kievit 1995). The apparatus is capable of measuring the size as well as the chemical composition, by the use of Laser Desorption/Ionisation (LDI), of an

  20. A novel magnet focusing plate for matrix-assisted laser desorption/ionization analysis of magnetic bead-bound analytes.

    Gode, David; Volmer, Dietrich A

    2013-05-15

    Magnetic beads are often used for serum profiling of peptide and protein biomarkers. In these assays, the bead-bound analytes are eluted from the beads prior to mass spectrometric analysis. This study describes a novel matrix-assisted laser desorption/ionization (MALDI) technique for direct application and focusing of magnetic beads to MALDI plates by means of dedicated micro-magnets as sample spots. Custom-made MALDI plates with magnetic focusing spots were made using small nickel-coated neodymium micro-magnets integrated into a stainless steel plate in a 16 × 24 (384) pattern. For demonstrating the proof-of-concept, commercial C-18 magnetic beads were used for the extraction of a test compound (reserpine) from aqueous solution. Experiments were conducted to study focusing abilities, the required laser energies, the influence of a matrix compound, dispensing techniques, solvent choice and the amount of magnetic beads. Dispensing the magnetic beads onto the micro-magnet sample spots resulted in immediate and strong binding to the magnetic surface. Light microscope images illustrated the homogeneous distribution of beads across the surfaces of the magnets, when the entire sample volume containing the beads was pipetted onto the surface. Subsequent MALDI analysis of the bead-bound analyte demonstrated excellent and reproducible ionization yields. The surface-assisted laser desorption/ionization (SALDI) properties of the strongly light-absorbing γ-Fe2O3-based beads resulted in similar ionization efficiencies to those obtained from experiments with an additional MALDI matrix compound. This feasibility study successfully demonstrated the magnetic focusing abilities for magnetic bead-bound analytes on a novel MALDI plate containing small micro-magnets as sample spots. One of the key advantages of this integrated approach is that no elution steps from magnetic beads were required during analyses compared with conventional bead experiments. Copyright © 2013 John Wiley

  1. Rapid Quantification of 25-Hydroxyvitamin D3 in Human Serum by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

    Qi, Yulin; Müller, Miriam; Stokes, Caroline S.; Volmer, Dietrich A.

    2018-04-01

    LC-MS/MS is widely utilized today for quantification of vitamin D in biological fluids. Mass spectrometric assays for vitamin D require very careful method optimization for precise and interference-free, accurate analyses however. Here, we explore chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) as a rapid alternative for quantitative measurement of 25-hydroxyvitamin D3 in human serum, and compare it to results from LC-MS/MS. The method implemented an automated imaging step of each MALDI spot, to locate areas of high intensity, avoid sweet spot phenomena, and thus improve precision. There was no statistically significant difference in vitamin D quantification between the MALDI-MS/MS and LC-MS/MS: mean ± standard deviation for MALDI-MS—29.4 ± 10.3 ng/mL—versus LC-MS/MS—30.3 ± 11.2 ng/mL (P = 0.128)—for the sum of the 25-hydroxyvitamin D epimers. The MALDI-based assay avoided time-consuming chromatographic separation steps and was thus much faster than the LC-MS/MS assay. It also consumed less sample, required no organic solvents, and was readily automated. In this proof-of-concept study, MALDI-MS readily demonstrated its potential for mass spectrometric quantification of vitamin D compounds in biological fluids.

  2. Analysis of Microbial Mixtures by Matrix-assisted Laser Desorption/Ionization time-of-flight Mass Spectrometry

    Wahl, Karen L.; Wunschel, Sharon C.; Jarman, Kristin H.; Valentine, Nancy B.; Petersen, Catherine E.; Kingsley, Mark T.; Zartolas, Kimberly A.; Saenz, Adam J.

    2002-12-15

    Many different laboratories are currently developing mass-spectrometric techniques to analyze and identify microorganisms. However, minimal work has been done with mixtures of bacteria. To demonstrate that microbial mixtures could be analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), mixed bacterial cultures were analyzed in a double-blind fashion. Nine different bacterial species currently in our MALDI-MS fingerprint library were used to generate 50 different simulated mixed bacterial cultures similar to that done for an initial blind study previously reported.(1) The samples were analyzed by MALDI-MS with automated data extraction and analysis algorithms developed in our laboratory. The components present in the sample were identified correctly to the species level in all but one of the samples. However, correctly eliminating closely related organisms was challenging for the current algorithms, especially in differentiating Serratia marcescens, Escherichia coli, and Yersinia enterocolitica, which have some similarities in their MALDI-MS fingerprints. Efforts to improve the specificity of the algorithms are in progress.

  3. Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry in Clinical Microbiology: What Are the Current Issues?

    van Belkum, Alex; Welker, Martin; Pincus, David; Charrier, Jean Philippe; Girard, Victoria

    2017-11-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has revolutionized the identification of microbial species in clinical microbiology laboratories. MALDI-TOF-MS has swiftly become the new gold-standard method owing to its key advantages of simplicity and robustness. However, as with all new methods, adoption of the MALDI-TOF MS approach is still not widespread. Optimal sample preparation has not yet been achieved for several applications, and there are continuing discussions on the need for improved database quality and the inclusion of additional microbial species. New applications such as in the field of antimicrobial susceptibility testing have been proposed but not yet translated to the level of ease and reproducibility that one should expect in routine diagnostic systems. Finally, during routine identification testing, unexpected results are regularly obtained, and the best methods for transmitting these results into clinical care are still evolving. We here discuss the success of MALDI-TOF MS in clinical microbiology and highlight fields of application that are still amenable to improvement. © The Korean Society for Laboratory Medicine.

  4. Ionization in matrix-assisted laser desorption/ionization: singly charged molecular ions are the lucky survivors.

    Karas, M; Glückmann, M; Schäfer, J

    2000-01-01

    A new model for the ionization processes in UV matrix-assisted laser desorption/ionization (MALDI) which accounts for the major phenomena observed is presented and discussed. The model retains elements of earlier approaches, such as photoionization and photochemical reactions, but it redefines these in the light of new working questions, most importantly why only singly charged ions are detected. Based on experimental evidence, the formation of singly and multiply charged clusters by a deficiency/excess of ions and also by photoionization and subsequent photochemical processes is pointed out to be the major ionization processes, which typically occur in parallel. The generation of electrons and their partial loss into the surrounding vacuum and solid, on the one hand, results in a positively charged ion-neutral plume facilitating a high overall ionization yield. On the other hand, these electrons, and also the large excess of protonated matrix ions in the negative ion mode, induce effective ion reneutralization in the plume. These neutralization processes are most effective for the highly charged cluster ions initially formed. Their fragmentation behaviour is evidenced in fast metastable fragmentation characteristics and agrees well with an electron capture dissociation mechanism and the enthalpy transfer upon neutralization forms the rationale for the prominent fragmentation and intense chemical noise accompanying successful MALDI. Within the course of the paper, cross-correlations with other desorption/ionization techniques and with earlier discussions on their mechanisms are drawn. Copyright 2000 John Wiley & Sons, Ltd.

  5. Identification and localization of trauma-related biomarkers using matrix assisted laser desorption/ionization imaging mass spectrometry

    Jones, Kirstin; Reilly, Matthew A.; Glickman, Randolph D.

    2017-02-01

    Current treatments for ocular and optic nerve trauma are largely ineffective and may have adverse side effects; therefore, new approaches are needed to understand trauma mechanisms. Identification of trauma-related biomarkers may yield insights into the molecular aspects of tissue trauma that can contribute to the development of better diagnostics and treatments. The conventional approach for protein biomarker measurement largely relies on immunoaffinity methods that typically can only be applied to analytes for which antibodies or other targeting means are available. Matrix assisted laser-assisted desorption/ionization imaging mass spectrometry (MALDI-IMS) is a specialized application of mass spectrometry that not only is well suited to the discovery of novel or unanticipated biomarkers, but also provides information about the spatial localization of biomarkers in tissue. We have been using MALDI-IMS to find traumarelated protein biomarkers in retina and optic nerve tissue from animal models subjected to ocular injury produced by either blast overpressure or mechanical torsion. Work to date by our group, using MALDI-IMS, found that the pattern of protein expression is modified in the injured ocular tissue as soon as 24 hr post-injury, compared to controls. Specific proteins may be up- or down-regulated by trauma, suggesting different tissue responses to a given injury. Ongoing work is directed at identifying the proteins affected and mapping their expression in the ocular tissue, anticipating that systematic analysis can be used to identify targets for prospective therapies for ocular trauma.

  6. Matrix-assisted laser desorption/ionization mass spectrometric analysis of aliphatic biodegradable photoluminescent polymers using new ionic liquid matrices.

    Serrano, Carlos A; Zhang, Yi; Yang, Jian; Schug, Kevin A

    2011-05-15

    In this study, two novel ionic liquid matrices (ILMs), N,N-diisopropylethylammonium 3-oxocoumarate and N,N-diisopropylethylammonium dihydroxymonooxoacetophenoate, were tested for the structural elucidation of recently developed aliphatic biodegradable polymers by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The polymers, formed by a condensation reaction of three components, citric acid, octane diol, and an amino acid, are fluorescent, but the exact mechanism behind their luminescent properties has not been fully elucidated. In the original studies, which introduced the polymer class (J. Yang et al., Proc. Natl. Acad. Sci. USA 2009, 106, 10086-10091), a hyper-conjugated cyclic structure was proposed as the source for the photoluminescent behavior. With the use of the two new ILMs, we present evidence that supports the presence of the proposed cyclization product. In addition, the new ILMs, when compared with a previously established ILM, N,N-diisopropylethylammonium α-cyano-3-hydroxycinnimate, provided similar signal intensities and maintained similar spectral profiles. This research also established that the new ILMs provided good spot-to-spot reproducibility and high ionization efficiency compared with corresponding crystalline matrix preparations. Many polymer features revealed through the use of the ILMs could not be observed with crystalline matrices. Ultimately, the new ILMs highlighted the composition of the synthetic polymers, as well as the loss of water that was expected for the formation of the proposed cyclic structure on the polymer backbone. Copyright © 2011 John Wiley & Sons, Ltd.

  7. Capsule Typing of Haemophilus influenzae by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry.

    Månsson, Viktor; Gilsdorf, Janet R; Kahlmeter, Gunnar; Kilian, Mogens; Kroll, J Simon; Riesbeck, Kristian; Resman, Fredrik

    2018-03-01

    Encapsulated Haemophilus influenzae strains belong to type-specific genetic lineages. Reliable capsule typing requires PCR, but a more efficient method would be useful. We evaluated capsule typing by using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Isolates of all capsule types (a-f and nontypeable; n = 258) and isogenic capsule transformants (types a-d) were investigated. Principal component and biomarker analyses of mass spectra showed clustering, and mass peaks correlated with capsule type-specific genetic lineages. We used 31 selected isolates to construct a capsule typing database. Validation with the remaining isolates (n = 227) showed 100% sensitivity and 92.2% specificity for encapsulated strains (a-f; n = 61). Blinded validation of a supplemented database (n = 50) using clinical isolates (n = 126) showed 100% sensitivity and 100% specificity for encapsulated strains (b, e, and f; n = 28). MALDI-TOF mass spectrometry is an accurate method for capsule typing of H. influenzae.

  8. Newborn screening by matrix-assisted laser desorption/ionization mass spectrometry based on parylene-matrix chip.

    Kim, Jo-Il; Noh, Joo-Yoon; Kim, Mira; Park, Jong-Min; Song, Hyun-Woo; Kang, Min-Jung; Pyun, Jae-Chul

    2017-08-01

    Newborn screening for diagnosis of phenylketonuria, homocystinuria, and maple syrup urine disease have been conducted by analyzing the concentration of target amino acids using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) based on parylene-matrix chip. Parylene-matrix chip was applied to MALDI-ToF MS analysis reducing the matrix peaks significantly at low mass-to-charge ratio range (m/z  0.98) and the LODs were ranging from 9.0 to 22.9 μg/mL. Effect of proteins in serum was estimated by comparing MALDI-ToF mass spectra of amino acids-spiked serum before and after the methanol extraction. Interference of other amino acids on analysis of target analyte was determined to be insignificant. From these results, MALDI-ToF MS based on parylene-matrix chip could be applicable to medical diagnosis of neonatal metabolic disorders. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Identification of Candida species isolated from vulvovaginitis using matrix assisted laser desorption ionization-time of flight mass spectrometry.

    Alizadeh, Majid; Kolecka, Anna; Boekhout, Teun; Zarrinfar, Hossein; Ghanbari Nahzag, Mohamad A; Badiee, Parisa; Rezaei-Matehkolaei, Ali; Fata, Abdolmajid; Dolatabadi, Somayeh; Najafzadeh, Mohammad J

    2017-12-01

    Vulvovaginal candidiasis (VVC) is a common problem in women. The purpose of this study was to identify Candida isolates by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) from women with vulvovaginitis that were referred to Ghaem Hospital, Mashhad, Iran. This study was conducted on 65 clinical samples isolated from women that were referred to Ghaem Hospital. All specimens were identified using phenotyping techniques, such as microscopy and culture on Sabouraud dextrose agar and corn meal agar. In addition, all isolates were processed for MALDI-TOF MS identification. Out of the 65 analyzed isolates, 61 (94%) samples were recognized by MALDI-TOF MS. However, the remaining four isolates (6%) had no reliable identification. According to the results, C. albicans (58.5%) was the most frequently isolated species, followed by C. tropicalis (16.9%), C. glabrata (7.7%), C. parapsilosis (7.7%), and guilliermondii (3.1%). As the findings indicated, MALDI TOF MS was successful in the identification of clinical Candida species. C. albicans was identified as the most common Candida species isolated from the women with VVC. Moreover, C. tropicalis was the most common species among the non- albicans Candida species.

  10. Detection of lung cancer using plasma protein profiling by matrix-assisted laser desorption/ionization mass spectrometry.

    Shevchenko, Valeriy E; Arnotskaya, Natalia E; Zaridze, David G

    2010-01-01

    There are no satisfactory plasma biomarkers which are available for the early detection and monitoring of lung cancer, one of the most frequent cancers worldwide. The aim of this study is to explore the application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) to plasma proteomic patterns to distinguish lung cancer patients from healthy individuals. The EDTA plasma samples have been pre-fractionated using magnetic bead kits functionalized with weak cation exchange coatings. We compiled MS protein profiles for 90 patients with squamous cell carcinomas (SCC) and compared them with profiles from 187 healthy controls. The MALDI-ToF spectra were analyzed statistically using ClinProTools bioinformatics software. Depending on the sample used, up to 441 peaks/spectrum could be detected in a mass range of 1000-20,000 Da; 33 of these proteins had statistically differential expression levels between SCC and control plasma (P 90%) in external validation test. These results suggest that plasma MALDI-ToF MS protein profiling can distinguish patients with SCC and also from healthy individuals with relatively high sensitivity and specificity and that MALDI- ToF MS is a potential tool for the screening of lung cancer.

  11. Rapid Identification of Intact Staphylococcal Bacteriophages Using Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry

    Dana Štveráková

    2018-04-01

    Full Text Available Staphylococcus aureus is a major causative agent of infections associated with hospital environments, where antibiotic-resistant strains have emerged as a significant threat. Phage therapy could offer a safe and effective alternative to antibiotics. Phage preparations should comply with quality and safety requirements; therefore, it is important to develop efficient production control technologies. This study was conducted to develop and evaluate a rapid and reliable method for identifying staphylococcal bacteriophages, based on detecting their specific proteins using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS profiling that is among the suggested methods for meeting the regulations of pharmaceutical authorities. Five different phage purification techniques were tested in combination with two MALDI-TOF MS matrices. Phages, either purified by CsCl density gradient centrifugation or as resuspended phage pellets, yielded mass spectra with the highest information value if ferulic acid was used as the MALDI matrix. Phage tail and capsid proteins yielded the strongest signals whereas the culture conditions had no effect on mass spectral quality. Thirty-seven phages from Myoviridae, Siphoviridae or Podoviridae families were analysed, including 23 siphophages belonging to the International Typing Set for human strains of S. aureus, as well as phages in preparations produced by Microgen, Bohemia Pharmaceuticals and MB Pharma. The data obtained demonstrate that MALDI-TOF MS can be used to effectively distinguish between Staphylococcus-specific bacteriophages.

  12. In Situ Analysis of Bacterial Lipopeptide Antibiotics by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.

    Debois, Delphine; Ongena, Marc; Cawoy, Hélène; De Pauw, Edwin

    2016-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) is a technique developed in the late 1990s enabling the two-dimensional mapping of a broad variety of biomolecules present at the surface of a sample. In many applications including pharmaceutical studies or biomarker discovery, the distribution of proteins, lipids or drugs, and metabolites may be visualized within tissue sections. More recently, MALDI MSI has become increasingly applied in microbiology where the versatility of the technique is perfectly suited to monitor the metabolic dynamics of bacterial colonies. The work described here is focused on the application of MALDI MSI to map secondary metabolites produced by Bacilli, especially lipopeptides, produced by bacterial cells during their interaction with their environment (bacteria, fungi, plant roots, etc.). This chapter addresses the advantages and challenges that the implementation of MALDI MSI to microbiological samples entails, including detailed protocols on sample preparation (from both microbiologist and mass spectrometrist points of view), matrix deposition, and data acquisition and interpretation. Lipopeptide images recorded from confrontation plates are also presented.

  13. Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry improves appropriateness of antibiotic treatment of bacteremia.

    Anne L M Vlek

    Full Text Available Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS allows the identification of microorganisms directly from positive blood culture broths. Use of the MALDI-TOF MS for rapid identification of microorganisms from blood culture broths can reduce the turnaround time to identification and may lead to earlier appropriate treatment of bacteremia. During February and April 2010, direct MALDI-TOF MS was routinely performed on all positive blood cultures. During December 2009 and March 2010 no direct MALDI-TOF MS was used. Information on antibiotic therapy was collected from the hospital and intensive care units' information systems from all positive blood cultures during the study period. In total, 253 episodes of bacteremia were included of which 89 during the intervention period and 164 during the control period. Direct performance of MALDI-TOF MS on positive blood culture broths reduced the time till species identification by 28.8-h and was associated with an 11.3% increase in the proportion of patients receiving appropriate antibiotic treatment 24 hours after blood culture positivity (64.0% in the control period versus 75.3% in the intervention period (p0.01. Routine implementation of this technique increased the proportion of patients on adequate antimicrobial treatment within 24 hours.

  14. Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry improves appropriateness of antibiotic treatment of bacteremia.

    Vlek, Anne L M; Bonten, Marc J M; Boel, C H Edwin

    2012-01-01

    Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows the identification of microorganisms directly from positive blood culture broths. Use of the MALDI-TOF MS for rapid identification of microorganisms from blood culture broths can reduce the turnaround time to identification and may lead to earlier appropriate treatment of bacteremia. During February and April 2010, direct MALDI-TOF MS was routinely performed on all positive blood cultures. During December 2009 and March 2010 no direct MALDI-TOF MS was used. Information on antibiotic therapy was collected from the hospital and intensive care units' information systems from all positive blood cultures during the study period. In total, 253 episodes of bacteremia were included of which 89 during the intervention period and 164 during the control period. Direct performance of MALDI-TOF MS on positive blood culture broths reduced the time till species identification by 28.8-h and was associated with an 11.3% increase in the proportion of patients receiving appropriate antibiotic treatment 24 hours after blood culture positivity (64.0% in the control period versus 75.3% in the intervention period (p0.01)). Routine implementation of this technique increased the proportion of patients on adequate antimicrobial treatment within 24 hours.

  15. Iron oxide nanomatrix facilitating metal ionization in matrix-assisted laser desorption/ionization mass spectrometry.

    Obena, Rofeamor P; Lin, Po-Chiao; Lu, Ying-Wei; Li, I-Che; del Mundo, Florian; Arco, Susan dR; Nuesca, Guillermo M; Lin, Chung-Chen; Chen, Yu-Ju

    2011-12-15

    The significance and epidemiological effects of metals to life necessitate the development of direct, efficient, and rapid method of analysis. Taking advantage of its simple, fast, and high-throughput features, we present a novel approach to metal ion detection by matrix-functionalized magnetic nanoparticle (matrix@MNP)-assisted MALDI-MS. Utilizing 21 biologically and environmentally relevant metal ion solutions, the performance of core and matrix@MNP against conventional matrixes in MALDI-MS and laser desorption ionization (LDI) MS were systemically tested to evaluate the versatility of matrix@MNP as ionization element. The matrix@MNPs provided 20- to >100-fold enhancement on detection sensitivity of metal ions and unambiguous identification through characteristic isotope patterns and accurate mass (<5 ppm), which may be attributed to its multifunctional role as metal chelator, preconcentrator, absorber, and reservoir of energy. Together with the comparison on the ionization behaviors of various metals having different ionization potentials (IP), we formulated a metal ionization mechanism model, alluding to the role of exciton pooling in matrix@MNP-assisted MALDI-MS. Moreover, the detection of Cu in spiked tap water demonstrated the practicability of this new approach as an efficient and direct alternative tool for fast, sensitive, and accurate determination of trace metal ions in real samples.

  16. The Effect of Collimating Lens Focusing on Laser Beam Shape in Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS).

    O'Rourke, Matthew B; Raymond, Benjamin B A; Djordjevic, Steven P; Padula, Matthew P

    2018-03-01

    Tissue imaging using matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a well-established technique that, in recent years, has seen wider adoption and novel application. Applications such imaging mass spectrometry (IMS) and biotyping are beginning to gain greater exposure and use; however, with limitations in optimization methods, producing the best result often relies on the ability to customize the physical characteristics of the instrumentation, a task that is challenging for most mass spectrometry laboratories. With this in mind, we have described the effect of making simple adjustments to the laser optics at the final collimating lens area, to adjust the laser beam size and shape in order to allow greater customization of the instrument for improving techniques such as IMS. We have therefore been able to demonstrate that improvements can be made without requiring the help of an electrical engineer or external funding in a way that only costs a small amount of time. Graphical Abstract ᅟ.

  17. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry: protocol standardization and database expansion for rapid identification of clinically important molds.

    Paul, Saikat; Singh, Pankaj; Rudramurthy, Shivaprakash M; Chakrabarti, Arunaloke; Ghosh, Anup K

    2017-12-01

    To standardize the matrix-assisted laser desorption ionization-time of flight mass spectrometry protocols and expansion of existing Bruker Biotyper database for mold identification. Four different sample preparation methods (protocol A, B, C and D) were evaluated. On analyzing each protein extraction method, reliable identification and best log scores were achieved through protocol D. The same protocol was used to identify 153 clinical isolates. Of these 153, 123 (80.3%) were accurately identified by using existing database and remaining 30 (19.7%) were not identified due to unavailability in database. On inclusion of missing main spectrum profile in existing database, all 153 isolates were identified. Matrix-assisted laser desorption ionization-time of flight mass spectrometry can be used for routine identification of clinically important molds.

  18. Performance of Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting

    Sleiman, Sue; Halliday, Catriona L.; Chapman, Belinda; Brown, Mitchell; Nitschke, Joanne; Lau, Anna F.

    2016-01-01

    We developed an Australian database for the identification of Aspergillus, Scedosporium, and Fusarium species (n = 28) by matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%; Fusarium, 84% versus 42%; and Scedosporium, 94% versus 18%, respectively). PMID:27252460

  19. Use of matrix-assisted laser desorption ionization-time of flight mass spectrometry for caspofungin susceptibility testing of Candida and Aspergillus species.

    De Carolis, Elena; Vella, Antonietta; Florio, Ada R; Posteraro, Patrizia; Perlin, David S; Sanguinetti, Maurizio; Posteraro, Brunella

    2012-07-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was evaluated for testing susceptibility to caspofungin of wild-type and fks mutant isolates of Candida and Aspergillus. Complete essential agreement was observed with the CLSI reference method, with categorical agreement for 94.1% of the Candida isolates tested. Thus, MALDI-TOF MS is a reliable and accurate method to detect fungal isolates with reduced caspofungin susceptibility.

  20. Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Caspofungin Susceptibility Testing of Candida and Aspergillus Species

    De Carolis, Elena; Vella, Antonietta; Florio, Ada R.; Posteraro, Patrizia; Perlin, David S.; Posteraro, Brunella

    2012-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) was evaluated for testing susceptibility to caspofungin of wild-type and fks mutant isolates of Candida and Aspergillus. Complete essential agreement was observed with the CLSI reference method, with categorical agreement for 94.1% of the Candida isolates tested. Thus, MALDI-TOF MS is a reliable and accurate method to detect fungal isolates with reduced caspofungin susceptibility. PMID:22535984

  1. Influence of Culture Media on Detection of Carbapenem Hydrolysis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Ramos, Ana Carolina; Carvalhaes, Cecília Godoy; Cordeiro-Moura, Jhonatha Rodrigo; Rockstroh, Anna Carolina; Machado, Antonia Maria Oliveira; Gales, Ana Cristina

    2016-07-01

    In this study, we evaluated the influence of distinct bacterial growth media on detection of carbapenemase hydrolysis by matrix-assisted laser desorption ionization-time of flight mass spectrometry. False-negative results were observed for OXA-25-, OXA-26-, and OXA-72-producing Acinetobacter baumannii isolates grown on MacConkey agar medium. The other culture media showed 100% sensitivity and 100% specificity for detecting carbapenemase. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  2. Natural products in Glycyrrhiza glabra (licorice) rhizome imaged at the cellular level by atmospheric pressure matrix-assisted laser desorption/ionization tandem mass spectrometry imaging

    Li, Bin; Bhandari, Dhaka Ram; Janfelt, Christian

    2014-01-01

    The rhizome of Glycyrrhiza glabra (licorice) was analyzed by high-resolution mass spectrometry imaging and tandem mass spectrometry imaging. An atmospheric pressure matrix-assisted laser desorption/ionization imaging ion source was combined with an orbital trapping mass spectrometer in order to o...... and saponins in legume species, combing the spatially resolved chemical information with morphological details at the microscopic level. Furthermore, the technique offers a scheme capable of high-throughput profiling of metabolites in plant tissues....

  3. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Direct Bacterial Identification from Positive Blood Culture Pellets ▿

    Prod'hom, Guy; Bizzini, Alain; Durussel, Christian; Bille, Jacques; Greub, Gilbert

    2010-01-01

    An ammonium chloride erythrocyte-lysing procedure was used to prepare a bacterial pellet from positive blood cultures for direct matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry analysis. Identification was obtained for 78.7% of the pellets tested. Moreover, 99% of the MALDI-TOF identifications were congruent at the species level when considering valid scores. This fast and accurate method is promising.

  4. Matrix-assisted laser desorption ionization-time of flight mass spectrometry for direct bacterial identification from positive blood culture pellets.

    Prod'hom, Guy; Bizzini, Alain; Durussel, Christian; Bille, Jacques; Greub, Gilbert

    2010-04-01

    An ammonium chloride erythrocyte-lysing procedure was used to prepare a bacterial pellet from positive blood cultures for direct matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry analysis. Identification was obtained for 78.7% of the pellets tested. Moreover, 99% of the MALDI-TOF identifications were congruent at the species level when considering valid scores. This fast and accurate method is promising.

  5. Matrix-Assisted Laser Desorption Ionization Mass Spectrometry of Compounds Containing Carboxyl Groups Using CdTe and CuO Nanoparticles

    Megumi Sakurai; Taro Sato; Jiawei Xu; Soichi Sato; Tatsuya Fujino

    2018-01-01

    Matrix-assisted laser desorption ionization mass spectrometry of compounds containing carboxyl groups was carried out by using semiconductor nanoparticles (CdTe and CuO) as the matrix. Salicylic acid (Sal), glucuronic acid (Glu), ibuprofen (Ibu), and tyrosine (Tyr) were ionized as deprotonated species (carboxylate anions) by using electrons ejected from CdTe after the photoexcitation. When CuO was used as the matrix, the peak intensity of Tyr became high compared with that obtained with CdTe....

  6. Rapid Identification of the Foodborne Pathogen Trichinella spp. by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Anne Mayer-Scholl

    Full Text Available Human trichinellosis occurs through consumption of raw or inadequately processed meat or meat products containing larvae of the parasitic nematodes of the genus Trichinella. Currently, nine species and three genotypes are recognized, of which T. spiralis, T. britovi and T. pseudospiralis have the highest public health relevance. To date, the differentiation of the larvae to the species and genotype level is based primarily on molecular methods, which can be relatively time consuming and labor intensive. Due to its rapidness and ease of use a matrix assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF MS reference spectra database using Trichinella strains of all known species and genotypes was created. A formicacid/acetonitrile protein extraction was carried out after pooling 10 larvae of each Trichinella species and genotype. Each sample was spotted 9 times using α-cyano 4-hydoxy cinnamic acid matrix and a MicroFlex LT mass spectrometer was used to acquire 3 spectra (m/z 2000 to 20000 Da from each spot resulting in 27 spectra/species or genotype. Following the spectra quality assessment, Biotyper software was used to create a main spectra library (MSP representing nine species and three genotypes of Trichinella. The evaluation of the spectra generated by MALDI-TOF MS revealed a classification which was comparable to the results obtained by molecular methods. Also, each Trichinella species utilized in this study was distinct and distinguishable with a high confidence level. Further, different conservation methods such as freezing and conservation in alcohol and the host species origin of the isolated larvae did not have a significant influence on the generated spectra. Therefore, the described MALDI-TOF MS can successfully be implemented for both genus and species level identification and represents a major step forward in the use of this technique in foodborne parasitology.

  7. Rapid screening of mixed edible oils and gutter oils by matrix-assisted laser desorption/ionization mass spectrometry

    Ng, Tsz-Tsun; So, Pui-Kin; Zheng, Bo [Food Safety and Technology Research Centre, State Key Laboratory of Chirosciences and Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom Kowloon, Hong Kong Special Administrative Region (China); Shenzhen Key Laboratory of Food Biological Safety Control and State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), Shenzhen Research Institute of The Hong Kong Polytechnic University, Shenzhen (China); Yao, Zhong-Ping, E-mail: zhongping.yao@polyu.edu.hk [Food Safety and Technology Research Centre, State Key Laboratory of Chirosciences and Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom Kowloon, Hong Kong Special Administrative Region (China); Shenzhen Key Laboratory of Food Biological Safety Control and State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), Shenzhen Research Institute of The Hong Kong Polytechnic University, Shenzhen (China)

    2015-07-16

    Highlights: • Simplified sample preparation method for direct analysis of edible oils by MALDI-MS. • Establishment of a preliminary MALDI-MS spectral database of edible oils. • Rapid screening of mixed edible oils and gutter oils. - Abstract: Authentication of edible oils is a long-term issue in food safety, and becomes particularly important with the emergence and wide spread of gutter oils in recent years. Due to the very high analytical demand and diversity of gutter oils, a high throughput analytical method and a versatile strategy for authentication of mixed edible oils and gutter oils are highly desirable. In this study, an improved matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) method has been developed for direct analysis of edible oils. This method involved on-target sample loading, automatic data acquisition and simple data processing. MALDI-MS spectra with high quality and high reproducibility have been obtained using this method, and a preliminary spectral database of edible oils has been set up. The authenticity of an edible oil sample can be determined by comparing its MALDI-MS spectrum and principal component analysis (PCA) results with those of its labeled oil in the database. This method is simple and the whole process only takes several minutes for analysis of one oil sample. We demonstrated that the method was sensitive to change in oil compositions and can be used for measuring compositions of mixed oils. The capability of the method for determining mislabeling enables it for rapid screening of gutter oils since fraudulent mislabeling is a common feature of gutter oils.

  8. Negative Ion In-Source Decay Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry for Sequencing Acidic Peptides

    McMillen, Chelsea L.; Wright, Patience M.; Cassady, Carolyn J.

    2016-05-01

    Matrix-assisted laser desorption/ionization (MALDI) in-source decay was studied in the negative ion mode on deprotonated peptides to determine its usefulness for obtaining extensive sequence information for acidic peptides. Eight biological acidic peptides, ranging in size from 11 to 33 residues, were studied by negative ion mode ISD (nISD). The matrices 2,5-dihydroxybenzoic acid, 2-aminobenzoic acid, 2-aminobenzamide, 1,5-diaminonaphthalene, 5-amino-1-naphthol, 3-aminoquinoline, and 9-aminoacridine were used with each peptide. Optimal fragmentation was produced with 1,5-diaminonphthalene (DAN), and extensive sequence informative fragmentation was observed for every peptide except hirudin(54-65). Cleavage at the N-Cα bond of the peptide backbone, producing c' and z' ions, was dominant for all peptides. Cleavage of the N-Cα bond N-terminal to proline residues was not observed. The formation of c and z ions is also found in electron transfer dissociation (ETD), electron capture dissociation (ECD), and positive ion mode ISD, which are considered to be radical-driven techniques. Oxidized insulin chain A, which has four highly acidic oxidized cysteine residues, had less extensive fragmentation. This peptide also exhibited the only charged localized fragmentation, with more pronounced product ion formation adjacent to the highly acidic residues. In addition, spectra were obtained by positive ion mode ISD for each protonated peptide; more sequence informative fragmentation was observed via nISD for all peptides. Three of the peptides studied had no product ion formation in ISD, but extensive sequence informative fragmentation was found in their nISD spectra. The results of this study indicate that nISD can be used to readily obtain sequence information for acidic peptides.

  9. Species Identification and Delineation of Pathogenic Mucorales by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Shao, Jin; Wan, Zhe; Li, Ruoyu; Yu, Jin

    2018-04-01

    This study aimed to validate the effectiveness of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based identification of filamentous fungi of the order Mucorales. A total of 111 isolates covering six genera preserved at the Research Center for Medical Mycology of Peking University were selected for MALDI-TOF MS analysis. We emphasized the study of 23 strains of Mucor irregularis predominantly isolated from patients in China. We first used the Bruker Filamentous Fungi library (v1.0) to identify all 111 isolates. To increase the identification rate, we created a compensatory in-house database, the Beijing Medical University (BMU) database, using 13 reference strains covering 6 species, including M. irregularis , Mucor hiemalis , Mucor racemosus , Cunninghamella bertholletiae , Cunninghamella phaeospora , and Cunninghamella echinulata All 111 isolates were then identified by MALDI-TOF MS using a combination of the Bruker library and BMU database. MALDI-TOF MS identified 55 (49.5%) and 74 (66.7%) isolates at the species and genus levels, respectively, using the Bruker Filamentous Fungi library v1.0 alone. A combination of the Bruker library and BMU database allowed MALDI-TOF MS to identify 90 (81.1%) and 111 (100%) isolates at the species and genus levels, respectively, with a significantly increased accuracy rate. MALDI-TOF MS poorly identified Mucorales when the Bruker library was used alone due to its lack of some fungal species. In contrast, this technique perfectly identified M. irregularis after main spectrum profiles (MSPs) of relevant reference strains were added to the Bruker library. With an expanded Bruker library, MALDI-TOF MS is an effective tool for the identification of pathogenic Mucorales. Copyright © 2018 American Society for Microbiology.

  10. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid identification of fungal rhinosinusitis pathogens.

    Huang, Yanfei; Wang, Jinglin; Zhang, Mingxin; Zhu, Min; Wang, Mei; Sun, Yufeng; Gu, Haitong; Cao, Jingjing; Li, Xue; Zhang, Shaoya; Lu, Xinxin

    2017-03-01

    Filamentous fungi are among the most important pathogens, causing fungal rhinosinusitis (FRS). Current laboratory diagnosis of FRS pathogens mainly relies on phenotypic identification by culture and microscopic examination, which is time consuming and expertise dependent. Although matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS has been employed to identify various fungi, its efficacy in the identification of FRS fungi is less clear. A total of 153 FRS isolates obtained from patients were analysed at the Clinical Laboratory at the Beijing Tongren Hospital affiliated to the Capital Medical University, between January 2014 and December 2015. They were identified by traditional phenotypic methods and Bruker MALDI-TOF MS (Bruker, Biotyper version 3.1), respectively. Discrepancies between the two methods were further validated by sequencing. Among the 153 isolates, 151 had correct species identification using MALDI-TOF MS (Bruker, Biot 3.1, score ≥2.0 or 2.3). MALDI-TOF MS enabled identification of some very closely related species that were indistinguishable by conventional phenotypic methods, including 1/10 Aspergillus versicolor, 3/20 Aspergillus flavus, 2/30 Aspergillus fumigatus and 1/20 Aspergillus terreus, which were misidentified by conventional phenotypic methods as Aspergillus nidulans, Aspergillus oryzae, Aspergillus japonicus and Aspergillus nidulans, respectively. In addition, 2/2 Rhizopus oryzae and 1/1 Rhizopus stolonifer that were identified only to the genus level by the phenotypic method were correctly identified by MALDI-TOF MS. MALDI-TOF MS is a rapid and accurate technique, and could replace the conventional phenotypic method for routine identification of FRS fungi in clinical microbiology laboratories.

  11. Microfluidic Isoelectric Focusing of Amyloid Beta Peptides Followed by Micropillar-Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry.

    Mikkonen, Saara; Jacksén, Johan; Roeraade, Johan; Thormann, Wolfgang; Emmer, Åsa

    2016-10-18

    A novel method for preconcentration and purification of the Alzheimer's disease related amyloid beta (Aβ) peptides by isoelectric focusing (IEF) in 75 nL microchannels combined with their analysis by micropillar-matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) is presented. A semiopen chip-based setup, consisting of open microchannels covered by a lid of a liquid fluorocarbon, was used. IEF was performed in a mixture of four small and chemically well-defined amphoteric carriers, glutamic acid, aspartyl-histidine (Asp-His), cycloserine (cSer), and arginine, which provided a stepwise pH gradient tailored for focusing of the C-terminal Aβ peptides with a pI of 5.3 in the boundary between cSer and Asp-His. Information about the focusing dynamics and location of the foci of Aβ peptides and other compounds was obtained using computer simulation and by performing MALDI-MS analysis directly from the open microchannel. With the established configuration, detection was performed by direct sampling of a nanoliter volume containing the focused Aβ peptides from the microchannel, followed by deposition of this volume onto a chip with micropillar MALDI targets. In addition to purification, IEF preconcentration provides at least a 10-fold increase of the MALDI-MS-signal. After immunoprecipitation and concentration of the eluate in the microchannel, IEF-micropillar-MALDI-MS is demonstrated to be a suitable platform for detection of Aβ peptides in human cerebrospinal fluid as well as in blood plasma.

  12. Microorganisms direct identification from blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Ferreira, L; Sánchez-Juanes, F; Porras-Guerra, I; García-García, M I; García-Sánchez, J E; González-Buitrago, J M; Muñoz-Bellido, J L

    2011-04-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows a fast and reliable bacterial identification from culture plates. Direct analysis of clinical samples may increase its usefulness in samples in which a fast identification of microorganisms can guide empirical treatment, such as blood cultures (BC). Three hundred and thirty BC, reported as positive by the automated BC incubation device, were processed by conventional methods for BC processing, and by a fast method based on direct MALDI-TOF MS. Three hundred and eighteen of them yield growth on culture plates, and 12 were false positive. The MALDI-TOF MS-based method reported that no peaks were found, or the absence of a reliable identification profile, in all these false positive BC. No mixed cultures were found. Among these 318 BC, we isolated 61 Gram-negatives (GN), 239 Gram-positives (GP) and 18 fungi. Microorganism identifications in GN were coincident with conventional identification, at the species level, in 83.3% of BC and, at the genus level, in 96.6%. In GP, identifications were coincident with conventional identification in 31.8% of BC at the species level, and in 64.8% at the genus level. Fungaemia was not reliably detected by MALDI-TOF. In 18 BC positive for Candida species (eight C. albicans, nine C. parapsilosis and one C. tropicalis), no microorganisms were identified at the species level, and only one (5.6%) was detected at the genus level. The results of the present study show that this fast, MALDI-TOF MS-based method allows bacterial identification directly from presumptively positive BC in a short time (<30 min), with a high accuracy, especially when GN bacteria are involved. © 2010 The Authors. Clinical Microbiology and Infection © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  13. Rapid screening of mixed edible oils and gutter oils by matrix-assisted laser desorption/ionization mass spectrometry

    Ng, Tsz-Tsun; So, Pui-Kin; Zheng, Bo; Yao, Zhong-Ping

    2015-01-01

    Highlights: • Simplified sample preparation method for direct analysis of edible oils by MALDI-MS. • Establishment of a preliminary MALDI-MS spectral database of edible oils. • Rapid screening of mixed edible oils and gutter oils. - Abstract: Authentication of edible oils is a long-term issue in food safety, and becomes particularly important with the emergence and wide spread of gutter oils in recent years. Due to the very high analytical demand and diversity of gutter oils, a high throughput analytical method and a versatile strategy for authentication of mixed edible oils and gutter oils are highly desirable. In this study, an improved matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) method has been developed for direct analysis of edible oils. This method involved on-target sample loading, automatic data acquisition and simple data processing. MALDI-MS spectra with high quality and high reproducibility have been obtained using this method, and a preliminary spectral database of edible oils has been set up. The authenticity of an edible oil sample can be determined by comparing its MALDI-MS spectrum and principal component analysis (PCA) results with those of its labeled oil in the database. This method is simple and the whole process only takes several minutes for analysis of one oil sample. We demonstrated that the method was sensitive to change in oil compositions and can be used for measuring compositions of mixed oils. The capability of the method for determining mislabeling enables it for rapid screening of gutter oils since fraudulent mislabeling is a common feature of gutter oils

  14. 1,8-Bis(dimethylamino)naphthalene/9-aminoacridine: A new binary matrix for lipid fingerprinting of intact bacteria by matrix assisted laser desorption ionization mass spectrometry

    Calvano, C.D., E-mail: cosimadamiana.calvano@uniba.it [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Monopoli, A.; Ditaranto, N. [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Palmisano, F. [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Centro Interdipartimentale di Ricerca S.M.A.R.T., Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy)

    2013-10-10

    Graphical abstract: -- Highlights: •New binary matrix for less ionizable lipid analysis with no interfering peaks. •Combined MALDI and X-ray photoelectron spectroscopy (XPS) analyses. •Fast lipid fingerprint on Gram positive and Gram negative bacteria by MALDI MS. •Mapping of phospholipids by XPS imaging. •Very fast membrane lipid extraction procedure. -- Abstract: The effectiveness of a novel binary matrix composed of 1,8-bis(dimethylamino)naphthalene (DMAN; proton sponge) and 9-aminoacridine (9AA) for the direct lipid analysis of whole bacterial cells by matrix assisted laser desorption ionization mass spectrometry (MALDI MS) is demonstrated. Deprotonated analyte signals nearly free of matrix-related ions were observed in negative ion mode. The effect of the most important factors (laser energy, pulse voltage, DMAN/9AA ratio, analyte/matrix ratio) was investigated using a Box–Behnken response surface design followed by multi-response optimization in order to simultaneously maximize signal-to-noise (S/N) ratio and resolution. The chemical surface composition of single or mixed matrices was explored by X-ray photoelectron spectroscopy (XPS). Moreover, XPS imaging was used to map the spatial distribution of a model phospholipid in single or binary matrices. The DMAN/9AA binary matrix was then successfully applied to the analysis of intact Gram positive (Lactobacillus sanfranciscensis) or Gram negative (Escherichia coli) microorganisms. About fifty major membrane components (free fatty acids, mono-, di- and tri-glycerides, phospholipids, glycolipids and cardiolipins) were quickly and easily detected over a mass range spanning from ca. 200 to ca. 1600 m/z. Moreover, mass spectra with improved S/N ratio (compared to single matrices), reduced chemical noise and no formation of matrix-clusters were invariably obtained demonstrating the potential of this binary matrix to improve sensitivity.

  15. 1,8-Bis(dimethylamino)naphthalene/9-aminoacridine: A new binary matrix for lipid fingerprinting of intact bacteria by matrix assisted laser desorption ionization mass spectrometry

    Calvano, C.D.; Monopoli, A.; Ditaranto, N.; Palmisano, F.

    2013-01-01

    Graphical abstract: -- Highlights: •New binary matrix for less ionizable lipid analysis with no interfering peaks. •Combined MALDI and X-ray photoelectron spectroscopy (XPS) analyses. •Fast lipid fingerprint on Gram positive and Gram negative bacteria by MALDI MS. •Mapping of phospholipids by XPS imaging. •Very fast membrane lipid extraction procedure. -- Abstract: The effectiveness of a novel binary matrix composed of 1,8-bis(dimethylamino)naphthalene (DMAN; proton sponge) and 9-aminoacridine (9AA) for the direct lipid analysis of whole bacterial cells by matrix assisted laser desorption ionization mass spectrometry (MALDI MS) is demonstrated. Deprotonated analyte signals nearly free of matrix-related ions were observed in negative ion mode. The effect of the most important factors (laser energy, pulse voltage, DMAN/9AA ratio, analyte/matrix ratio) was investigated using a Box–Behnken response surface design followed by multi-response optimization in order to simultaneously maximize signal-to-noise (S/N) ratio and resolution. The chemical surface composition of single or mixed matrices was explored by X-ray photoelectron spectroscopy (XPS). Moreover, XPS imaging was used to map the spatial distribution of a model phospholipid in single or binary matrices. The DMAN/9AA binary matrix was then successfully applied to the analysis of intact Gram positive (Lactobacillus sanfranciscensis) or Gram negative (Escherichia coli) microorganisms. About fifty major membrane components (free fatty acids, mono-, di- and tri-glycerides, phospholipids, glycolipids and cardiolipins) were quickly and easily detected over a mass range spanning from ca. 200 to ca. 1600 m/z. Moreover, mass spectra with improved S/N ratio (compared to single matrices), reduced chemical noise and no formation of matrix-clusters were invariably obtained demonstrating the potential of this binary matrix to improve sensitivity

  16. Semiconductor Nanomaterials-Based Fluorescence Spectroscopic and Matrix-Assisted Laser Desorption/Ionization (MALDI Mass Spectrometric Approaches to Proteome Analysis

    Suresh Kumar Kailasa

    2013-12-01

    Full Text Available Semiconductor quantum dots (QDs or nanoparticles (NPs exhibit very unusual physico-chemcial and optical properties. This review article introduces the applications of semiconductor nanomaterials (NMs in fluorescence spectroscopy and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS for biomolecule analysis. Due to their unique physico-chemical and optical properties, semiconductors NMs have created many new platforms for investigating biomolecular structures and information in modern biology. These semiconductor NMs served as effective fluorescent probes for sensing proteins and cells and acted as affinity or concentrating probes for enriching peptides, proteins and bacteria proteins prior to MALDI-MS analysis.

  17. Identification of barley and rye varieties using matrix- assisted laser desorption/ionisation time-of-flight mass spectrometry with neural networks

    Bloch, H.A.; Petersen, Marianne Kjerstine; Sperotto, Maria Maddalena

    2001-01-01

    developed, which combines analysis of alcohol-soluble wheat proteins (gliadins) using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry with neural networks. Here we have applied the same method for the identification of both barley (Hordeum vulgare L.) and rye (Secale cereale L.......) varieties. For barley, 95% of the mass spectra were correctly classified. This is an encouraging result, since in earlier experiments only a grouping into subsets of varieties was possible. However, the method was not useful in the classification of rye, due to the strong similarity between mass spectra...

  18. Performance of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting.

    Sleiman, Sue; Halliday, Catriona L; Chapman, Belinda; Brown, Mitchell; Nitschke, Joanne; Lau, Anna F; Chen, Sharon C-A

    2016-08-01

    We developed an Australian database for the identification of Aspergillus, Scedosporium, and Fusarium species (n = 28) by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%; Fusarium, 84% versus 42%; and Scedosporium, 94% versus 18%, respectively). Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  19. Detection and quantification of neurotensin in human brain tissue by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Gobom, J; Kraeuter, K O; Persson, R

    2000-01-01

    A method was developed for mass spectrometric detection of neurotensin (NT)-like immunoreactivity and quantification of NT in human brain tissue. The method is based on immunoprecipitation followed by analysis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF......-MS). The identity of the major component of the immunoprecipitates as neurotensin was confirmed by fragment ion analysis on an electrospray ionization quadrupole time-of-flight instrument. MALDI-TOF-MS quantification of NT was achieved using stable-isotope-labeled NT as the internal standard, yielding an error...

  20. Species identification of clinical isolates of anaerobic bacteria: a comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry systems

    Justesen, Ulrik Stenz; Holm, Anette; Knudsen, Elisa

    2011-01-01

    We compared two matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) systems (Shimadzu/SARAMIS and Bruker) on a collection of consecutive clinically important anaerobic bacteria (n = 290). The Bruker system had more correct identifications to the species level...... (67.2% versus 49.0%), but also more incorrect identifications (7.9% versus 1.4%). The system databases need to be optimized to increase identification levels. However, MALDI-TOF MS in its present version seems to be a fast and inexpensive method for identification of most clinically important...

  1. Laser pulse stacking method

    Moses, E.I.

    1992-12-01

    A laser pulse stacking method is disclosed. A problem with the prior art has been the generation of a series of laser beam pulses where the outer and inner regions of the beams are generated so as to form radially non-synchronous pulses. Such pulses thus have a non-uniform cross-sectional area with respect to the outer and inner edges of the pulses. The present invention provides a solution by combining the temporally non-uniform pulses in a stacking effect to thus provide a more uniform temporal synchronism over the beam diameter. 2 figs.

  2. Measurement of laser activated electron tunneling from semiconductor zinc oxide to adsorbed organic molecules by a matrix assisted laser desorption ionization mass spectrometer

    Zhong Hongying; Fu Jieying; Wang Xiaoli; Zheng Shi

    2012-01-01

    Highlights: ► Irradiation of photons with energies more than the band gap generates electron–hole pairs. ► Electron tunneling probability is dependent on the electron mobility. ► Tunneling electrons are captured by charge deficient atoms. ► Unpaired electrons induce cleavages of chemical bonds. - Abstract: Measurement of light induced heterogeneous electron transfer is important for understanding of fundamental processes involved in chemistry, physics and biology, which is still challenging by current techniques. Laser activated electron tunneling (LAET) from semiconductor metal oxides was observed and characterized by a MALDI (matrix assisted laser desorption ionization) mass spectrometer in this work. Nanoparticles of ZnO were placed on a MALDI sample plate. Free fatty acids and derivatives were used as models of organic compounds and directly deposited on the surface of ZnO nanoparticles. Irradiation of UV laser (λ = 355 nm) with energy more than the band gap of ZnO produces ions that can be detected in negative mode. When TiO 2 nanoparticles with similar band gap but much lower electron mobility were used, these ions were not observed unless the voltage on the sample plate was increased. The experimental results indicate that laser induced electron tunneling is dependent on the electron mobility and the strength of the electric field. Capture of low energy electrons by charge-deficient atoms of adsorbed organic molecules causes unpaired electron-directed cleavages of chemical bonds in a nonergodic pathway. In positive detection mode, electron tunneling cannot be observed due to the reverse moving direction of electrons. It should be able to expect that laser desorption ionization mass spectrometry is a new technique capable of probing the dynamics of electron tunneling. LAET offers advantages as a new ionization dissociation method for mass spectrometry.

  3. Ultrashort Laser Pulse Phenomena

    Diels, Jean-Claude

    2006-01-01

    Ultrashort Laser Pulse Phenomena, 2e serves as an introduction to the phenomena of ultra short laser pulses and describes how this technology can be used to examine problems in areas such as electromagnetism, optics, and quantum mechanics. Ultrashort Laser Pulse Phenomena combines theoretical backgrounds and experimental techniques and will serve as a manual on designing and constructing femtosecond (""faster than electronics"") systems or experiments from scratch. Beyond the simple optical system, the various sources of ultrashort pulses are presented, again with emphasis on the basic

  4. Short communication: Identification of subclinical cow mastitis pathogens in milk by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Barreiro, J R; Ferreira, C R; Sanvido, G B; Kostrzewa, M; Maier, T; Wegemann, B; Böttcher, V; Eberlin, M N; dos Santos, M V

    2010-12-01

    Subclinical mastitis is a common and easily disseminated disease in dairy herds. Its routine diagnosis via bacterial culture and biochemical identification is a difficult and time-consuming process. In this work, we show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows bacterial identification with high confidence and speed (1 d for bacterial growth and analysis). With the use of MALDI-TOF MS, 33 bacterial culture isolates from milk of different dairy cows from several farms were analyzed, and the results were compared with those obtained by classical biochemical methods. This proof-of-concept case demonstrates the reliability of MALDI-TOF MS bacterial identification, and its increased selectivity as illustrated by the additional identification of coagulase-negative Staphylococcus species and mixed bacterial cultures. Matrix-assisted laser desorption-ionization mass spectrometry considerably accelerates the diagnosis of mastitis pathogens, especially in cases of subclinical mastitis. More immediate and efficient animal management strategies for mastitis and milk quality control in the dairy industry can therefore be applied. Copyright © 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  5. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a new possibility for the identification and typing of anaerobic bacteria.

    Nagy, Elizabeth

    2014-01-01

    Anaerobic bacteria predominate in the normal flora of humans and are important, often life-threatening pathogens in mixed infections originating from the indigenous microbiota. The isolation and identification of anaerobes by phenotypic and DNA-based molecular methods at a species level is time-consuming and laborious. Following the successful adaptation of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the routine laboratory identification of bacteria, the extensive development of a database has been initiated to use this method for the identification of anaerobic bacteria. Not only frequently isolated anaerobic species, but also newly recognized and taxonomically rearranged genera and species can be identified using direct smear samples or whole-cell protein extraction, and even phylogenetically closely related species can be identified correctly by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Typing of anaerobic bacteria on a subspecies level, determination of antibiotic resistance and direct identification of blood culture isolates will revolutionize anaerobe bacteriology in the near future.

  6. Rapid identification of pathogens directly from blood culture bottles by Bruker matrix-assisted laser desorption laser ionization-time of flight mass spectrometry versus routine methods.

    Jamal, Wafaa; Saleem, Rola; Rotimi, Vincent O

    2013-08-01

    The use of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification of microorganisms directly from blood culture is an exciting dimension to the microbiologists. We evaluated the performance of Bruker SepsiTyper kit™ (STK) for direct identification of bacteria from positive blood culture. This was done in parallel with conventional methods. Nonrepetitive positive blood cultures from 160 consecutive patients were prospectively evaluated by both methods. Of 160 positive blood cultures, the STK identified 114 (75.6%) isolates and routine conventional method 150 (93%). Thirty-six isolates were misidentified or not identified by the kit. Of these, 5 had score of >2.000 and 31 had an unreliable low score of <1.7. Four of 8 yeasts were identified correctly. The average turnaround time using the STK was 35 min, including extraction steps and 30:12 to 36:12 h with routine method. The STK holds promise for timely management of bacteremic patients. Copyright © 2013 Elsevier Inc. All rights reserved.

  7. Correlation between phosphorylation ratios by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis and radioactivities by radioactive assay.

    Tsuchiya, Akira; Asai, Daisuke; Kang, Jeong-Hun; Mori, Takeshi; Niidome, Takuro; Katayama, Yoshiki

    2012-02-15

    To investigate the correlation between the counts per minute (CPM) by radioactivity assay and the phosphorylation ratio by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, we prepared 136 peptide substrates. The correlation coefficient of phosphorylation ratios to CPM was 0.77 for all samples. However, the more the numbers of positively charged amino acids increased, the more the correlation coefficient increased. Although positively charged amino acids can have an effect on the correlation results, MALDI-TOF MS analysis is a useful means for monitoring phosphorylated peptide and protein kinase activity instead of radioactivity assays. Copyright © 2011 Elsevier Inc. All rights reserved.

  8. Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of clinically important yeast species.

    Stevenson, Lindsay G; Drake, Steven K; Shea, Yvonne R; Zelazny, Adrian M; Murray, Patrick R

    2010-10-01

    We evaluated the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the rapid identification of yeast species. Using Bruker Daltonics MALDI BioTyper software, we created a spectral database library with m/z ratios of 2,000 to 20,000 Da for 109 type and reference strains of yeast (44 species in 8 genera). The database was tested for accuracy by use of 194 clinical isolates (23 species in 6 genera). A total of 192 (99.0%) of the clinical isolates were identified accurately by MALDI-TOF MS. The MALDI-TOF MS-based method was found to be reproducible and accurate, with low consumable costs and minimal preparation time.

  9. Detection and Quantification of 4-Methylimidazole in Cola by Matrix-assisted Laser Desorption Ionization Mass Spectrometry with Fe2O3 Nanoparticles on Zeolite.

    Fujii, Yosuke; Ding, Yuqi; Umezawa, Taichi; Akimoto, Takafumi; Xu, Jiawei; Uchida, Takashi; Fujino, Tatsuya

    2018-01-01

    Food additives generally used in carbonated drinks, such as 4-methylimidazole (4MI), caffeine (Caf?), citric acid (CA), and aspartame (Apm), were measured by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) using nanometer-sized particles of iron oxide (Fe 2 O 3 NPs). The quantification of 4MI in Coca Cola (C-cola) was carried out. In order to improve the reproducibility of the peak intensities, Fe 2 O 3 NPs loaded on ZSM5 zeolite were used as the matrix for quantification. By using 2-ethylimidazole (2EI) as the internal standard, the amount of 4MI in C-cola was determined to range from 88 to 65 μg/355 mL. The results agree with the published value (approx. 72 μg/355 mL). It was found that MALDI using Fe 2 O 3 was applicable to the quantification of 4MI in C-cola.

  10. [Separation and identification of bovine lactoferricin by high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/ time of flight mass spectrometry].

    An, Meichen; Liu, Ning

    2010-02-01

    A high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (HPLC-MALDI-TOF/TOF MS) method was developed for the separation and identification of bovine lactoferricin (LfcinB). Bovine lactoferrin was hydrolyzed by pepsin and then separated by ion exchange chromatography and reversed-phase liquid chromatography (RP-LC). The antibacterial activities of the fractions from RP-LC separation were determined and the protein concentration of the fraction with the highest activity was measured, whose sequence was indentified by MALDI-TOF/TOF MS. The relative molecular mass of LfcinB was 3 124.89 and the protein concentration was 18.20 microg/mL. The method of producing LfcinB proposed in this study has fast speed, high accuracy and high resolution.

  11. Metabolomic Analysis of Oxidative and Glycolytic Skeletal Muscles by Matrix-Assisted Laser Desorption/IonizationMass Spectrometric Imaging (MALDI MSI)

    Tsai, Yu-Hsuan; Garrett, Timothy J.; Carter, Christy S.; Yost, Richard A.

    2015-06-01

    Skeletal muscles are composed of heterogeneous muscle fibers that have different physiological, morphological, biochemical, and histological characteristics. In this work, skeletal muscles extensor digitorum longus, soleus, and whole gastrocnemius were analyzed by matrix-assisted laser desorption/ionization mass spectrometry to characterize small molecule metabolites of oxidative and glycolytic muscle fiber types as well as to visualize biomarker localization. Multivariate data analysis such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed to extract significant features. Different metabolic fingerprints were observed from oxidative and glycolytic fibers. Higher abundances of biomolecules such as antioxidant anserine as well as acylcarnitines were observed in the glycolytic fibers, whereas taurine and some nucleotides were found to be localized in the oxidative fibers.

  12. Composite glycerol/graphite/aromatic acid matrices for thin-layer chromatography/matrix-assisted laser desorption/ionization mass spectrometry of heterocyclic compounds.

    Esparza, Cesar; Borisov, R S; Varlamov, A V; Zaikin, V G

    2016-10-28

    New composite matrices have been suggested for the analysis of mixtures of different synthetic organic compounds (N-containing heterocycles and erectile dysfunction drugs) by thin layer chromatography/matrix-assisted laser desorption ionization time-of-flight mass spectrometry (TLC/MALDI-TOF). Different mixtures of classical MALDI matrices and graphite particles dispersed in glycerol were used for the registration of MALDI mass spectra directly from TLC plates after analytes separation. In most of cases, the mass spectra possessed [M+H] + ions; however, for some analytes only [M+Na] + and [M+K] + ions were observed. These ions have been used to generate visualized TLC chromatograms. The described approach increases the desorption/ionization efficiencies of analytes separated by TLC, prevent spot blurring, simplifies and decrease time for sample preparation. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures

    Kjellström, Sven; Jensen, Ole Nørregaard

    2003-01-01

    A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic...... matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS enabled the detection of the hydrophobic membrane protein bacteriorhodopsin as a component in a simple protein mixture. Peptide mixtures containing...... phosphorylated, glycosylated, or acylated peptides were successfully separated and analyzed by the in situ LLE-MALDI MS technique and demonstrate the potential of this method for enhanced separation and structural analysis of posttranslationally modified peptides in proteomics research....

  14. Reproducibility of serum protein profiling by systematic assessment using solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry

    Callesen, Anne K; Christensen, René Depont; Madsen, Jonna S

    2008-01-01

    for serum protein profiling we investigated a range of sample preparation techniques and developed a statistical method based on repeated analyses for evaluation of protein-profiling performance of MALDI MS. Two different solid-phase extraction (SPE) methods were investigated, namely custom......Protein profiling of human serum by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) is potentially a new diagnostic tool for early detection of human diseases, including cancer. Sample preparation is a key issue in MALDI MS and the analysis of complex samples such as serum......-made microcolumns and commercially available magnetic beads. Using these two methods, nineteen different sample preparation methods for serum profiling by MALDI MS were systematically tested with regard to matrix selection, stationary phase, selectivity, and reproducibility. Microcolumns were tested with regard...

  15. Hippocampal lipid differences in Alzheimer's disease: a human brain study using matrix-assisted laser desorption/ionization-imaging mass spectrometry.

    Mendis, Lakshini H S; Grey, Angus C; Faull, Richard L M; Curtis, Maurice A

    2016-10-01

    Alzheimer's disease (AD), the leading cause of dementia, is pathologically characterized by β-amyloid plaques and tau tangles. However, there is also evidence of lipid dyshomeostasis-mediated AD pathology. Given the structural diversity of lipids, mass spectrometry is a useful tool for studying lipid changes in AD. Although there have been a few studies investigating lipid changes in the human hippocampus in particular, there are few reports on how lipids change in each hippocampal subfield (e.g., Cornu Ammonis [CA] 1-4, dentate gyrus [DG] etc.). Since each subfield has its own function, we postulated that there could be lipid changes that are unique to each. We used matrix-assisted laser desorption/ionization-imaging mass spectrometry to investigate specific lipid changes in each subfield in AD. Data from the hippocampus region of six age- and gender-matched normal and AD pairs were analyzed with SCiLS lab 2015b software (SCiLS GmbH, Germany; RRID:SCR_014426), using an analysis workflow developed in-house. Hematoxylin, eosin, and luxol fast blue staining were used to precisely delineate each anatomical hippocampal subfield. Putative lipid identities, which were consistent with published data, were assigned using MS/MS. Both positively and negatively charged lipid ion species were abundantly detected in normal and AD tissue. While the distribution pattern of lipids did not change in AD, the abundance of some lipids changed, consistent with trends that have been previously reported. However, our results indicated that the majority of these lipid changes specifically occur in the CA1 region. Additionally, there were many lipid changes that were specific to the DG. Matrix-assisted laser desorption/ionization-imaging mass spectrometry and our analysis workflow provide a novel method to investigate specific lipid changes in hippocampal subfields. Future work will focus on elucidating the role that specific lipid differences in each subfield play in AD pathogenesis.

  16. Mesoporous tungsten titanate as matrix for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of biomolecules

    Shan Zhe; Han Lu; Yuan Minjia; Deng Chunhui; Zhao Dongyuan; Tu Bo; Yang Pengyuan

    2007-01-01

    In this paper, mesoporous tungsten titanate (WTiO) with different nano-pore structures was utilized as matrix for the analysis of short peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). Effect of characteristic features of mesoporous matrices on laser desorption/ionization process was investigated. Experiments showed that the ordered two-dimensional and three-dimensional mesoporous matrices were superior in performance to the non-ordered WTiO matrix. The dramatic enhancement of signal sensitivity by the ordered mesoporous matrices can be reasonably attributed to the ordered structure, which facilitated the understanding on structure-function relationship in mesoporous cavity for laser desorption process of adsorbed biomolecules. With the ordered mesoporous matrix, the short peptides are successfully detected. The presence of trace alkali metal salt effectively increased the analyte ion yields and the MALDI-TOFMS using the inorganic mesoporous matrices displayed a high salt tolerance. The developed technique also showed a satisfactory performance in peptide-mapping and amino-acid sequencing analysis

  17. Water ice as a matrix for film production by matrix-assisted pulsed laser evaporation (MAPLE)

    Rodrigo, K; Schou, J; Toftmann, B; Pedrys, R

    2007-01-01

    We have studied water ice as a matrix for the production of PEG (polyethylene glycol) films by MAPLE at 355 nm. The deposition rate is small compared with other matrices typically used in MAPLE, but the deposition of photofragments from the matrix can be avoided. At temperatures above -50deg. C of the target holder the deposition rate increases strongly, but the evaporation pressure in the MAPLE chamber also increases drastically

  18. Water ice as a matrix for film production by matrix assisted pulsed laser evaporation (MAPLE)

    Rodrigo, Katarzyna Agnieszka; Schou, Jørgen; Christensen, Bo Toftmann

    2007-01-01

    We have studied water ice as a matrix for the production of PEG (polyethylene glycol) films by MAPLE at 355 nm. The deposition rate is small compared with other matrices typically used in MAPLE, but the deposition of photofragments from the matrix can be avoided. At temperatures above -50 degrees C...... of the target holder the deposition rate increases strongly, but the evaporation pressure in the MAPLE chamber also increases drastically....

  19. Independent assessment of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) sample preparation quality : Effect of sample preparation on MALDI-MS of synthetic polymers

    Kooijman, Pieter C.; Kok, Sander; Honing, Maarten

    2017-01-01

    Rationale: Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) provides detailed and in-depth information about the molecular characteristics of synthetic polymers. To obtain the most accurate results the sample preparation parameters should be chosen to suit the sample and the

  20. Localization of an O-glycosylated site in the recombinant barley alpha-amylase 1 produced in yeast and correction of the amino acid sequence using matrix-assisted laser desorption/ionization mass spectrometry of peptide mixtures

    Andersen, Jens S.; Søgaard, M; Svensson, B

    1994-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of peptide mixtures was used to characterize recombinant barley alpha-amylase 1, produced in yeast. Three peptide mixtures were generated by cleavage with CNBr, digestion with endoproteinase Lys-C and Asp-N, respectively...

  1. Classification of wheat varieties: Use of two-dimensional gel electrophoresis for varieties that can not be classified by matrix assisted laser desorption/ionization-time of flight-mass spectrometry and an artificial neural network

    Jacobsen, Susanne; Nesic, Ljiljana; Petersen, Marianne Kjerstine

    2001-01-01

    Analyzing a gliadin extract by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI- TOF-MS) combined with an artificial neural network (ANN) is a suitable method for identification of wheat varieties. However, the ANN can not distinguish between all different wheat...

  2. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Analysis of Gram-Positive, Catalase-Negative Cocci Not Belonging to the Streptococcus or Enterococcus Genus and Benefits of Database Extension

    Christensen, Jens Jørgen; Dargis, Rimtas; Hammer, Monja

    2012-01-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry with a Bruker Daltonics microflex LT system was applied to 90 well-characterized catalase-negative, Gram-positive cocci not belonging to the streptococci or enterococci. Biotyper version 2.0.43.1 software...

  3. The importance of matrix-assisted laser desorption ionization–time of flight mass spectrometry for correct identification of Clostridium difficile isolated from chromID C. difficile chromogenic agar

    Jonathan H.K. Chen

    2017-10-01

    Full Text Available The clinical workflow of using chromogenic agar and matrix-assisted laser desorption ionization time-of-fight mass spectrometry (MALDI-TOF MS for Clostridium difficile identification was evaluated. The addition of MALDI-TOF MS identification after the chromID C. difficile chromogenic agar culture could significantly improve the diagnostic accuracy of C. difficile.

  4. Pulsed inductive HF laser

    Razhev, A M; Kargapol' tsev, E S [Institute of Laser Physics, Siberian Branch, Russian Academy of Sciences, Novosibirsk (Russian Federation); Churkin, D S; Demchuk, S V [Novosibirsk State University, Novosibirsk (Russian Federation)

    2016-03-31

    We report the results of experimentally investigated dependences of temporal, spectral and spatial characteristics of an inductive HF-laser generation on the pump conditions. Gas mixtures H{sub 2} – F{sub 2}(NF{sub 3} or SF6{sub 6}) and He(Ne) – H{sub 2} – F{sub 2}(NF{sub 3} or SF{sub 6}) were used as active media. The FWHM pulse duration reached 0.42 μs. This value corresponded to a pulsed power of 45 kW. For the first time, the emission spectrum of an inductive HF laser was investigated, which consisted of seven groups of bands with centres around the wavelengths of 2732, 2736, 2739, 2835, 2837, 2893 and 2913 nm. The cross section profile of the laser beam was a ring with a diameter of about 20 mm and width of about 5 mm. Parameters of laser operation in the repetitively pulsed regime were sufficiently stable. The amplitude instability of light pulses was no greater than 5% – 6%. (lasers)

  5. Pulsed atomic soliton laser

    Carr, L.D.; Brand, J.

    2004-01-01

    It is shown that simultaneously changing the scattering length of an elongated, harmonically trapped Bose-Einstein condensate from positive to negative and inverting the axial portion of the trap, so that it becomes expulsive, results in a train of self-coherent solitonic pulses. Each pulse is itself a nondispersive attractive Bose-Einstein condensate that rapidly self-cools. The axial trap functions as a waveguide. The solitons can be made robustly stable with the right choice of trap geometry, number of atoms, and interaction strength. Theoretical and numerical evidence suggests that such a pulsed atomic soliton laser can be made in present experiments

  6. Fundamental study of hydrogen-attachment-induced peptide fragmentation occurring in the gas phase and during the matrix-assisted laser desorption/ionization process.

    Asakawa, Daiki; Takahashi, Hidenori; Iwamoto, Shinichi; Tanaka, Koichi

    2018-05-09

    Mass spectrometry with hydrogen-radical-mediated fragmentation techniques has been used for the sequencing of proteins/peptides. The two methods, matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) and hydrogen attachment/abstraction dissociation (HAD) are known as hydrogen-radical-mediated fragmentation techniques. MALDI-ISD occurs during laser induced desorption processes, whereas HAD utilizes the association of hydrogen with peptide ions in the gas phase. In this study, the general mechanisms of MALDI-ISD and HAD of peptides were investigated. We demonstrated the fragmentation of four model peptides and investigated the fragment formation pathways using density functional theory (DFT) calculations. The current experimental and computational joint study indicated that MALDI-ISD and HAD produce aminoketyl radical intermediates, which immediately undergo radical-induced cleavage at the N-Cα bond located on the C-terminal side of the radical site, leading to the c'/z˙ fragment pair. In the case of MALDI-ISD, the z˙ fragments undergo a subsequent reaction with the matrix to give z' and matrix adducts of the z fragments. In contrast, the c' and z˙ fragments react with hydrogen atoms during the HAD processes, and various fragment species, such as c˙, c', z˙ and z', were observed in the HAD-MS/MS mass spectra.

  7. Differentiation of Lactobacillus brevis strains using Matrix-Assisted-Laser-Desorption-Ionization-Time-of-Flight Mass Spectrometry with respect to their beer spoilage potential.

    Kern, Carola C; Vogel, Rudi F; Behr, Jürgen

    2014-06-01

    Lactobacillus (L.) brevis is one of the most frequently encountered bacteria in beer-spoilage incidents. As the species Lactobacillus brevis comprises strains showing varying ability to grow in beer, ranging from growth in low hopped wheat to highly hopped pilsner beer, differentiation and classification of L. brevis with regard to their beer-spoiling ability is of vital interest for the brewing industry. Matrix-Assisted-Laser-Desorption-Ionization-Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) has been shown as a powerful tool for species and sub-species differentiation of bacterial isolates and is increasingly used for strain-level differentiation. Seventeen L. brevis strains, representative of different spoilage types, were characterized according to their tolerance to iso-alpha-acids and their growth in wheat-, lager- and pilsner beer. MALDI-TOF MS spectra were acquired to perform strain-level identification, cluster analysis and biomarker detection. Strain-level identification was achieved in 90% out of 204 spectra. Misidentification occurred nearly exclusively among strains belonging to the same spoilage type. Though spectra of strongly beer-spoiling strains showed remarkable similarity, no decisive single markers were detected to be present in all strains of one group. However, MALDI-TOF MS spectra can be reliably assigned to the corresponding strain and thus allow to track single strains and connect them to their physiological properties. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Multi-imaging of Cytokinin and Abscisic Acid on the Roots of Rice (Oryza sativa) Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Shiono, Katsuhiro; Hashizaki, Riho; Nakanishi, Toyofumi; Sakai, Tatsuko; Yamamoto, Takushi; Ogata, Koretsugu; Harada, Ken-Ichi; Ohtani, Hajime; Katano, Hajime; Taira, Shu

    2017-09-06

    Plant hormones act as important signaling molecules that regulate responses to abiotic stress as well as plant growth and development. Because their concentrations of hormones control the physiological responses in the target tissue, it is important to know the distributions and concentrations in the tissues. However, it is difficult to determine the hormone concentration on the plant tissue as a result of the limitations of conventional methods. Here, we report the first multi-imaging of two plant hormones, one of cytokinin [i.e., trans-zeatin (tZ)] and abscisic acid (ABA) using a new technology, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) imaging. Protonated signals of tZ (m/z 220.1) and ABA (m/z 265.3) were chosen on longitudinal sections of rice roots for MS imaging. tZ was broadly distributed about 40 mm behind the root apex but was barely detectable at the apex, whereas ABA was mainly detected at the root apex. Multi-imaging using MALDI-TOF-MS enabled the visualization of the localization and quantification of plant hormones. Thus, this tool is applicable to a wide range of plant species growing under various environmental conditions.

  9. Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry.

    Krismer, Jasmin; Sobek, Jens; Steinhoff, Robert F; Fagerer, Stephan R; Pabst, Martin; Zenobi, Renato

    2015-08-15

    The consequences of cellular heterogeneity, such as biocide persistence, can only be tackled by studying each individual in a cell population. Fluorescent tags provide tools for the high-throughput analysis of genomes, RNA transcripts, or proteins on the single-cell level. However, the analysis of lower-molecular-weight compounds that elude tagging is still a great challenge. Here, we describe a novel high-throughput microscale sample preparation technique for single cells that allows a mass spectrum to be obtained for each individual cell within a microbial population. The approach presented includes spotting Chlamydomonas reinhardtii cells, using a noncontact microarrayer, onto a specialized slide and controlled lysis of cells separated on the slide. Throughout the sample preparation, analytes were traced and individual steps optimized using autofluorescence detection of chlorophyll. The lysates of isolated cells are subjected to a direct, label-free analysis using matrix-assisted laser desorption ionization mass spectrometry. Thus, we were able to differentiate individual cells of two Chlamydomonas reinhardtii strains based on single-cell mass spectra. Furthermore, we showed that only population profiles with real single-cell resolution render a nondistorted picture of the phenotypes contained in a population. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. A survey of useful salt additives in matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry of lipids: introducing nitrates for improved analysis.

    Griffiths, Rian L; Bunch, Josephine

    2012-07-15

    Matrix-assisted laser desorption/ionization (MALDI) is a powerful technique for the direct analysis of lipids in complex mixtures and thin tissue sections, making it an extremely attractive method for profiling lipids in health and disease. Lipids are readily detected as [M+H](+), [M+Na](+) and [M+K](+) ions in positive ion MALDI mass spectrometry (MS) experiments. This not only decreases sensitivity, but can also lead to overlapping m/z values of the various adducts of different lipids. Additives can be used to promote formation of a particular adduct, improving sensitivity, reducing spectral complexity and enhancing structural characterization in collision-induced dissociation (CID) experiments. Li(+), Na(+), K(+), Cs(+) and NH(4)(+) cations were considered as a range of salt types (acetates, chlorides and nitrates) incorporated into DHB matrix solutions at concentrations between 5 and 80 mM. The study was extended to evaluate the effect of these additives on CID experiments of a lipid standard, after optimization of collision energy parameters. Experiments were performed on a hybrid quadrupole time-of-flight (QqTOF) instrument. The systematic evaluation of new and existing additives in MALDI-MS and MS/MS of lipids demonstrated the importance of additive cation and anion choice and concentration for tailoring spectral results. The recommended choice of additive depends on the desired outcomes of the experiment to be performed (MS or MS/MS). Nitrates are found to be particularly useful additives for lipid analysis. Copyright © 2012 John Wiley & Sons, Ltd.

  11. Proteogenomic biomarkers for identification of Francisella species and subspecies by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

    Durighello, Emie; Bellanger, Laurent; Ezan, Eric; Armengaud, Jean

    2014-10-07

    Francisella tularensis is the causative agent of tularemia. Because some Francisella strains are very virulent, this species is considered by the Centers for Disease Control and Prevention to be a potential category A bioweapon. A mass spectrometry method to quickly and robustly distinguish between virulent and nonvirulent Francisella strains is desirable. A combination of shotgun proteomics and whole-cell matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry on the Francisella tularensis subsp. holarctica LVS defined three protein biomarkers that allow such discrimination: the histone-like protein HU form B, the 10 kDa chaperonin Cpn10, and the 50S ribosomal protein L24. We established that their combined detection by whole-cell MALDI-TOF spectrum could enable (i) the identification of Francisella species, and (ii) the prediction of their virulence level, i.e., gain of a taxonomical level with the identification of Francisella tularensis subspecies. The detection of these biomarkers by MALDI-TOF mass spectrometry is straightforward because of their abundance and the absence of other abundant protein species closely related in terms of m/z. The predicted molecular weights for the three biomarkers and their presence as intense peaks were confirmed with MALDI-TOF/MS spectra acquired on Francisella philomiragia ATCC 25015 and on Francisella tularensis subsp. tularensis CCUG 2112, the most virulent Francisella subspecies.

  12. Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

    Jeong, Youngsu; Lee, Jonghee; Kim, Seongsoo

    2013-01-01

    The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field

  13. Evaluation of a simple protein extraction method for species identification of clinically relevant staphylococci by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Matsuda, Naoto; Matsuda, Mari; Notake, Shigeyuki; Yokokawa, Hirohide; Kawamura, Yoshiaki; Hiramatsu, Keiichi; Kikuchi, Ken

    2012-12-01

    In clinical microbiology, bacterial identification is labor-intensive and time-consuming. A solution for this problem is the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In this study, we evaluated a modified protein extraction method of identification performed on target plates (on-plate extraction method) with MALDI-TOF (Bruker Microflex LT with Biotyper version 3.0) and compared it to 2 previously described methods: the direct colony method and a standard protein extraction method (standard extraction method). We evaluated the species of 273 clinical strains and 14 reference strains of staphylococci. All isolates were characterized using the superoxide dismutase A sequence as a reference. For the species identification, the on-plate, standard extraction, and direct colony methods identified 257 isolates (89.5%), 232 isolates (80.8%), and 173 isolates (60.2%), respectively, with statistically significant differences among the three methods (P extraction method is at least as good as standard extraction in identification rate and has the advantage of a shorter processing time.

  14. Peptidylation for the determination of low-molecular-weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Tang, Feng; Cen, Si-Ying; He, Huan; Liu, Yi; Yuan, Bi-Feng; Feng, Yu-Qi

    2016-05-23

    Determination of low-molecular-weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has been a great challenge in the analytical research field. Here we developed a universal peptide-based derivatization (peptidylation) strategy for the sensitive analysis of low-molecular-weight compounds by MALDI-TOF-MS. Upon peptidylation, the molecular weights of target analytes increase, thus avoiding serious matrix ion interference in the low-molecular-weight region in MALDI-TOF-MS. Since peptides typically exhibit good signal response during MALDI-TOF-MS analysis, peptidylation endows high detection sensitivities of low-molecular-weight analytes. As a proof-of-concept, we analyzed low-molecular-weight compounds of aldehydes and thiols by the developed peptidylation strategy. Our results showed that aldehydes and thiols can be readily determined upon peptidylation, thus realizing the sensitive and efficient determination of low-molecular-weight compounds by MALDI-TOF-MS. Moreover, target analytes also can be unambiguously detected in biological samples using the peptidylation strategy. The established peptidylation strategy is a universal strategy and can be extended to the sensitive analysis of various low-molecular-weight compounds by MALDI-TOF-MS, which may be potentially used in areas such as metabolomics.

  15. Quantitative analysis of polyhexamethylene guanidine (PHMG) oligomers via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with an ionic-liquid matrix.

    Yoon, Donhee; Lee, Dongkun; Lee, Jong-Hyeon; Cha, Sangwon; Oh, Han Bin

    2015-01-30

    Quantifying polymers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) with a conventional crystalline matrix generally suffers from poor sample-to-sample or shot-to-shot reproducibility. An ionic-liquid matrix has been demonstrated to mitigate these reproducibility issues by providing a homogeneous sample surface, which is useful for quantifying polymers. In the present study, we evaluated the use of an ionic liquid matrix, i.e., 1-methylimidazolium α-cyano-4-hydroxycinnamate (1-MeIm-CHCA), to quantify polyhexamethylene guanidine (PHMG) samples that impose a critical health hazard when inhaled in the form of droplets. MALDI-TOF mass spectra were acquired for PHMG oligomers using a variety of ionic-liquid matrices including 1-MeIm-CHCA. Calibration curves were constructed by plotting the sum of the PHMG oligomer peak areas versus PHMG sample concentration with a variety of peptide internal standards. Compared with the conventional crystalline matrix, the 1-MeIm-CHCA ionic-liquid matrix had much better reproducibility (lower standard deviations). Furthermore, by using an internal peptide standard, good linear calibration plots could be obtained over a range of PMHG concentrations of at least 4 orders of magnitude. This study successfully demonstrated that PHMG samples can be quantitatively characterized by MALDI-TOFMS with an ionic-liquid matrix and an internal standard. Copyright © 2014 John Wiley & Sons, Ltd.

  16. Recent advances in matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) for in situ analysis of endogenous molecules in plants.

    Qin, Liang; Zhang, Yawen; Liu, Yaqin; He, Huixin; Han, Manman; Li, Yanyan; Zeng, Maomao; Wang, Xiaodong

    2018-04-17

    Mass spectrometry imaging (MSI) as a label-free and powerful imaging technique enables in situ evaluation of a tissue metabolome and/or proteome, becoming increasingly popular in the detection of plant endogenous molecules. The characterization of structure and spatial information of endogenous molecules in plants are both very important aspects to better understand the physiological mechanism of plant organism. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) is a commonly-used tissue imaging technique, which requires matrix to assist in situ detection of a variety of molecules on the surface of a tissue section. In previous studies, MALDI-MSI was mostly used for the detection of molecules from animal tissue sections, compared to plant samples due to cell structural limitations, such as plant cuticles, epicuticular waxes, and cell walls. Despite the enormous progress that has been made in tissue imaging, there is still a challenge for MALDI-MSI suitable for the imaging of endogenous compounds in plants. This review summarises the recent advances in MALDI-MSI, focusing on the application of in situ detection of endogenous molecules in different plant organs, i.e. root, stem, leaf, flower, fruit, and seed. Further improvements on instrumentation sensitivity, matrix selection, image processing and sample preparation will expand the application of MALDI-MSI in plant research. Copyright © 2018 John Wiley & Sons, Ltd.

  17. Flexible xxx-asp/asn and gly-xxx residues of equine cytochrome C in matrix-assisted laser desorption/ionization in-source decay mass spectrometry.

    Takayama, Mitsuo

    2012-01-01

    The backbone flexibility of a protein has been studied from the standpoint of the susceptibility of amino acid residues to in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Residues more susceptible to MALDI-ISD, namely Xxx-Asp/Asn and Gly-Xxx, were identified from the discontinuous intense peak of c'-ions originating from specific cleavage at N-Cα bonds of the backbone of equine cytochrome c. The identity of the residues susceptible to ISD was consistent with the known flexible backbone amides as estimated by hydrogen/deuterium exchange (HDX) experiments. The identity of these flexible amino acid residues (Asp, Asn, and Gly) is consistent with the fact that these residues are preferred in flexible secondary structure free from intramolecular hydrogen-bonded structures such as α-helix and β-sheet. The MALDI-ISD spectrum of equine cytochrome c gave not only intense N-terminal side c'-ions originating from N-Cα bond cleavage at Xxx-Asp/Asn and Gly-Xxx residues, but also C-terminal side complement z'-ions originating from the same cleavage sites. The present study implies that MALDI-ISD can give information about backbone flexibility of proteins, comparable with the protection factors estimated by HDX.

  18. Flexible Xxx–Asp/Asn and Gly–Xxx Residues of Equine Cytochrome c in Matrix-Assisted Laser Desorption/Ionization In-Source Decay Mass Spectrometry

    Takayama, Mitsuo

    2012-01-01

    The backbone flexibility of a protein has been studied from the standpoint of the susceptibility of amino acid residues to in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Residues more susceptible to MALDI-ISD, namely Xxx–Asp/Asn and Gly–Xxx, were identified from the discontinuous intense peak of c′-ions originating from specific cleavage at N–Cα bonds of the backbone of equine cytochrome c. The identity of the residues susceptible to ISD was consistent with the known flexible backbone amides as estimated by hydrogen/deuterium exchange (HDX) experiments. The identity of these flexible amino acid residues (Asp, Asn, and Gly) is consistent with the fact that these residues are preferred in flexible secondary structure free from intramolecular hydrogen-bonded structures such as α-helix and β-sheet. The MALDI-ISD spectrum of equine cytochrome c gave not only intense N-terminal side c′-ions originating from N–Cα bond cleavage at Xxx–Asp/Asn and Gly–Xxx residues, but also C-terminal side complement z′-ions originating from the same cleavage sites. The present study implies that MALDI-ISD can give information about backbone flexibility of proteins, comparable with the protection factors estimated by HDX. PMID:24349908

  19. Discrimination of Aspergillus isolates at the species and strain level by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry fingerprinting.

    Hettick, Justin M; Green, Brett J; Buskirk, Amanda D; Kashon, Michael L; Slaven, James E; Janotka, Erika; Blachere, Francoise M; Schmechel, Detlef; Beezhold, Donald H

    2008-09-15

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to generate highly reproducible mass spectral fingerprints for 12 species of fungi of the genus Aspergillus and 5 different strains of Aspergillus flavus. Prior to MALDI-TOF MS analysis, the fungi were subjected to three 1-min bead beating cycles in an acetonitrile/trifluoroacetic acid solvent. The mass spectra contain abundant peaks in the range of 5 to 20kDa and may be used to discriminate between species unambiguously. A discriminant analysis using all peaks from the MALDI-TOF MS data yielded error rates for classification of 0 and 18.75% for resubstitution and cross-validation methods, respectively. If a subset of 28 significant peaks is chosen, resubstitution and cross-validation error rates are 0%. Discriminant analysis of the MALDI-TOF MS data for 5 strains of A. flavus using all peaks yielded error rates for classification of 0 and 5% for resubstitution and cross-validation methods, respectively. These data indicate that MALDI-TOF MS data may be used for unambiguous identification of members of the genus Aspergillus at both the species and strain levels.

  20. Hydrophilic interaction chromatography coupled matrix assisted laser desorption/ionization mass spectrometry for molecular analysis of organic compounds in medicines, tea, and coffee.

    Wang, Ren-Qi; Bao, Kai; Croué, Jean-Philippe; Ng, Siu Choon

    2013-11-21

    Natural occurring organic compounds from food, natural organic matter, as well as metabolic products have received intense attention in current chemical and biological studies. Examination of unknown compounds in complex sample matrices is hampered by the limited choices for data readout and molecular elucidation. Herein, we report a generic method of hydrophilic interaction chromatography (HILIC) coupled with matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) for the rapid characterization of ingredients in pharmaceutical compounds, tea, and coffee. The analytes were first fractionated using a cationic HILIC column prior to MALDI-MS analyses. It was found that the retention times of a compound arising from different samples were consistent under the same conditions. Accordingly, molecules can be readily characterized by both the mass and chromatographic retention time. The retention behaviors of acidic and basic compounds on the cationic HILIC column were found to be significantly influenced by the pH of mobile phases, whereas neutral compounds depicted a constant retention time at different pH. The general HILIC-MALDI-MS method is feasible for fast screening of naturally occurring organic compounds. A series of homologs can be determined if they have the same retention behavior. Their structural features can be elucidated by considering their mass differences and hydrophilic properties as determined by HILIC chromatogram.

  1. Visualization of dynamic change in contraction-induced lipid composition in mouse skeletal muscle by matrix-assisted laser desorption/ionization imaging mass spectrometry.

    Goto-Inoue, Naoko; Manabe, Yasuko; Miyatake, Shouta; Ogino, Shinya; Morishita, Ai; Hayasaka, Takahiro; Masaki, Noritaka; Setou, Mitsutoshi; Fujii, Nobuharu L

    2012-06-01

    Lipids in skeletal muscle play a fundamental role both in normal muscle metabolism and in disease states. Skeletal muscle lipid accumulation is associated with several chronic metabolic disorders, including obesity, insulin resistance, and type 2 diabetes. However, it is poorly understood whether the lipid composition of skeletal muscle changes by contraction, due to the complexity of lipid molecular species. In this study, we used matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) to investigate changes in skeletal muscle lipid composition induced by contraction. We successfully observed the reduction of diacylglycerol and triacylglycerol, which are generally associated with muscle contraction. Interestingly, we found the accumulation of some saturated and mono-unsaturated fatty acids and poly-unsaturated fatty acids containing phosphatidylcholine in contracted muscles. Moreover, the distributions of several types of lipid were changed by contraction. Our results show that changes in the lipid amount, lipid composition, and energy metabolic activity can be evaluated in each local spot of cells and tissues at the same time using MALDI-IMS. In conclusion, MALDI-IMS is a powerful tool for studying lipid changes associated with contractions.

  2. Evaluation of synthase and hemisynthase activities of glucosamine-6-phosphate synthase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Gaucher-Wieczorek, Florence; Guérineau, Vincent; Touboul, David; Thétiot-Laurent, Sophie; Pelissier, Franck; Badet-Denisot, Marie-Ange; Badet, Bernard; Durand, Philippe

    2014-08-01

    Glucosamine-6-phosphate synthase (GlmS, EC 2.6.1.16) catalyzes the first and rate-limiting step in the hexosamine biosynthetic pathway, leading to the synthesis of uridine-5'-diphospho-N-acetyl-D-glucosamine, the major building block for the edification of peptidoglycan in bacteria, chitin in fungi, and glycoproteins in mammals. This bisubstrate enzyme converts D-fructose-6-phosphate (Fru-6P) and L-glutamine (Gln) into D-glucosamine-6-phosphate (GlcN-6P) and L-glutamate (Glu), respectively. We previously demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) allows determination of the kinetic parameters of the synthase activity. We propose here to refine the experimental protocol to quantify Glu and GlcN-6P, allowing determination of both hemisynthase and synthase parameters from a single assay kinetic experiment, while avoiding interferences encountered in other assays. It is the first time that MALDI-MS is used to survey the activity of a bisubstrate enzyme. Copyright © 2014 Elsevier Inc. All rights reserved.

  3. Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

    Clark, Andrew E.; Kaleta, Erin J.; Arora, Amit

    2013-01-01

    SUMMARY Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the “nuts and bolts” of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care. PMID:23824373

  4. Matrix-assisted laser desorption ionization-time of flight mass spectrometry: a fundamental shift in the routine practice of clinical microbiology.

    Clark, Andrew E; Kaleta, Erin J; Arora, Amit; Wolk, Donna M

    2013-07-01

    Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the "nuts and bolts" of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care.

  5. Feasibility of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) networking in university hospitals in Brussels.

    Martiny, D; Cremagnani, P; Gaillard, A; Miendje Deyi, V Y; Mascart, G; Ebraert, A; Attalibi, S; Dediste, A; Vandenberg, O

    2014-05-01

    The mutualisation of analytical platforms might be used to address rising healthcare costs. Our study aimed to evaluate the feasibility of networking a unique matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) system for common use in several university hospitals in Brussels, Belgium. During a one-month period, 1,055 successive bacterial isolates from the Brugmann University Hospital were identified on-site using conventional techniques; these same isolates were also identified using a MALDI-TOF MS system at the Porte de Hal Laboratory by sending target plates and identification projects via transportation and the INFECTIO_MALDI software (Infopartner, Nancy, France), respectively. The occurrence of transmission problems (MS networking always provided a faster identification result than conventional techniques, except when chromogenic culture media and oxidase tests were used (p MS networking could lead to substantial annual cost savings. MALDI-TOF MS networking presents many advantages, and few conventional techniques (optochin and oxidase tests) are required to ensure the same quality in patient care from the distant laboratory. Nevertheless, such networking should not be considered unless there is a reorganisation of workflow, efficient communication between teams, qualified technologists and a reliable IT department and helpdesk to manage potential connectivity problems.

  6. Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.

    Ayyadurai, Saravanan; Flaudrops, Christophe; Raoult, Didier; Drancourt, Michel

    2010-11-12

    Accurate identification is necessary to discriminate harmless environmental Yersinia species from the food-borne pathogens Yersinia enterocolitica and Yersinia pseudotuberculosis and from the group A bioterrorism plague agent Yersinia pestis. In order to circumvent the limitations of current phenotypic and PCR-based identification methods, we aimed to assess the usefulness of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) protein profiling for accurate and rapid identification of Yersinia species. As a first step, we built a database of 39 different Yersinia strains representing 12 different Yersinia species, including 13 Y. pestis isolates representative of the Antiqua, Medievalis and Orientalis biotypes. The organisms were deposited on the MALDI-TOF plate after appropriate ethanol-based inactivation, and a protein profile was obtained within 6 minutes for each of the Yersinia species. When compared with a 3,025-profile database, every Yersinia species yielded a unique protein profile and was unambiguously identified. In the second step of analysis, environmental and clinical isolates of Y. pestis (n = 2) and Y. enterocolitica (n = 11) were compared to the database and correctly identified. In particular, Y. pestis was unambiguously identified at the species level, and MALDI-TOF was able to successfully differentiate the three biotypes. These data indicate that MALDI-TOF can be used as a rapid and accurate first-line method for the identification of Yersinia isolates.

  7. Multivariate analysis of matrix-assisted laser desorption/ionization mass spectrometric data related to glycoxidation products of human globins in nephropathic patients.

    Lapolla, Annunziata; Ragazzi, Eugenio; Andretta, Barbara; Fedele, Domenico; Tubaro, Michela; Seraglia, Roberta; Molin, Laura; Traldi, Pietro

    2007-06-01

    To clarify the possible pathogenetic role of oxidation products originated from the glycation of proteins, human globins from nephropathic patients have been studied by matrix-assisted laser desorption/ionization mass spectrometry (MALDI), revealing not only unglycated and monoglycated globins, but also a series of different species. For the last ones, structural assignments were tentatively done on the basis of observed masses and expectations for the Maillard reaction pattern. Consequently, they must be considered only propositive, and the discussion which will follow must be considered in this view. In our opinion this approach does not seem to compromise the intended diagnostic use of the data because distinctions are valid even if the assignments are uncertain. We studied nine healthy subjects and 19 nephropathic patients and processed the data obtained from the MALDI spectra using a multivariate analysis. Our results showed that multivariate analytical techniques enable differential aspects of the profile of molecular species to be identified in the blood of end stage nephropathic patients. A correct grouping can be achieved by principal component analysis (PCA) and the results suggest that several products involved in carbonyl stress exist in nephropathic patients. These compounds may have a relevant role as specific markers of the pathological state.

  8. Identification of Cronobacter species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with an optimized analysis method.

    Wang, Qi; Zhao, Xiao-Juan; Wang, Zi-Wei; Liu, Li; Wei, Yong-Xin; Han, Xiao; Zeng, Jing; Liao, Wan-Jin

    2017-08-01

    Rapid and precise identification of Cronobacter species is important for foodborne pathogen detection, however, commercial biochemical methods can only identify Cronobacter strains to genus level in most cases. To evaluate the power of mass spectrometry based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF MS) for Cronobacter species identification, 51 Cronobacter strains (eight reference and 43 wild strains) were identified by both MALDI-TOF MS and 16S rRNA gene sequencing. Biotyper RTC provided by Bruker identified all eight reference and 43 wild strains as Cronobacter species, which demonstrated the power of MALDI-TOF MS to identify Cronobacter strains to genus level. However, using the Bruker's database (6903 main spectra products) and Biotyper software, the MALDI-TOF MS analysis could not identify the investigated strains to species level. When MALDI-TOF MS analysis was performed using the combined in-house Cronobacter database and Bruker's database, bin setting, and unweighted pair group method with arithmetic mean (UPGMA) clustering, all the 51 strains were clearly identified into six Cronobacter species and the identification accuracy increased from 60% to 100%. We demonstrated that MALDI-TOF MS was reliable and easy-to-use for Cronobacter species identification and highlighted the importance of establishing a reliable database and improving the current data analysis methods by integrating the bin setting and UPGMA clustering. Copyright © 2017. Published by Elsevier B.V.

  9. Decision peptide-driven: a free software tool for accurate protein quantification using gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry.

    Santos, Hugo M; Reboiro-Jato, Miguel; Glez-Peña, Daniel; Nunes-Miranda, J D; Fdez-Riverola, Florentino; Carvallo, R; Capelo, J L

    2010-09-15

    The decision peptide-driven tool implements a software application for assisting the user in a protocol for accurate protein quantification based on the following steps: (1) protein separation through gel electrophoresis; (2) in-gel protein digestion; (3) direct and inverse (18)O-labeling and (4) matrix assisted laser desorption ionization time of flight mass spectrometry, MALDI analysis. The DPD software compares the MALDI results of the direct and inverse (18)O-labeling experiments and quickly identifies those peptides with paralleled loses in different sets of a typical proteomic workflow. Those peptides are used for subsequent accurate protein quantification. The interpretation of the MALDI data from direct and inverse labeling experiments is time-consuming requiring a significant amount of time to do all comparisons manually. The DPD software shortens and simplifies the searching of the peptides that must be used for quantification from a week to just some minutes. To do so, it takes as input several MALDI spectra and aids the researcher in an automatic mode (i) to compare data from direct and inverse (18)O-labeling experiments, calculating the corresponding ratios to determine those peptides with paralleled losses throughout different sets of experiments; and (ii) allow to use those peptides as internal standards for subsequent accurate protein quantification using (18)O-labeling. In this work the DPD software is presented and explained with the quantification of protein carbonic anhydrase. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  10. Influence of secondary structure on in-source decay of protein in matrix-assisted laser desorption/ionization mass spectrometry.

    Takayama, Mitsuo; Osaka, Issey; Sakakura, Motoshi

    2012-01-01

    The susceptibility of the N-Cα bond of the peptide backbone to specific cleavage by in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) was studied from the standpoint of the secondary structure of three proteins. A naphthalene derivative, 5-amino-1-naphtol (5,1-ANL), was used as the matrix. The resulting c'-ions, which originate from the cleavage at N-Cα bonds in flexible secondary structures such as turn and bend, and are free from intra-molecular hydrogen-bonded α-helix structure, gave relatively intense peaks. Furthermore, ISD spectra of the proteins showed that the N-Cα bonds of specific amino acid residues, namely Gly-Xxx, Xxx-Asp, and Xxx-Asn, were more susceptible to MALDI-ISD than other amino acid residues. This is in agreement with the observation that Gly, Asp and Asn residues usually located in turns, rather than α-helix. The results obtained indicate that protein molecules embedded into the matrix crystal in the MALDI experiments maintain their secondary structures as determined by X-ray crystallography, and that MALDI-ISD has the capability for providing information concerning the secondary structure of protein.

  11. Evaluation of matrix-assisted laser desorption/ionization time of flight mass spectrometry for the identification of ceratopogonid and culicid larvae.

    Steinmann, I C; Pflüger, V; Schaffner, F; Mathis, A; Kaufmann, C

    2013-03-01

    Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was evaluated for the rapid identification of ceratopogonid larvae. Optimal sample preparation as evaluated with laboratory-reared biting midges Culicoides nubeculosus was the homogenization of gut-less larvae in 10% formic acid, and analysis of 0.2 mg/ml crude protein homogenate mixed with SA matrix at a ratio of 1:1.5. Using 5 larvae each of 4 ceratopogonid species (C. nubeculosus, C. obsoletus, C. decor, and Dasyhelea sp.) and of 2 culicid species (Aedes aegypti, Ae. japonicus), biomarker mass sets between 27 and 33 masses were determined. In a validation study, 67 larvae belonging to the target species were correctly identified by automated database-based identification (91%) or manual full comparison (9%). Four specimens of non-target species did not yield identification. As anticipated for holometabolous insects, the biomarker mass sets of adults cannot be used for the identification of larvae, and vice versa, because they share only very few similar masses as shown for C. nubeculosus, C. obsoletus, and Ae. japonicus. Thus, protein profiling by MALDI-TOF as a quick, inexpensive and accurate alternative tool is applicable to identify insect larvae of vector species collected in the field.

  12. Ammonium Sulfate Improves Detection of Hydrophilic Quaternary Ammonium Compounds through Decreased Ion Suppression in Matrix-Assisted Laser Desorption/Ionization Imaging Mass Spectrometry.

    Sugiyama, Eiji; Masaki, Noritaka; Matsushita, Shoko; Setou, Mitsutoshi

    2015-11-17

    Hydrophilic quaternary ammonium compounds (QACs) include derivatives of carnitine (Car) or choline, which are known to have essential bioactivities. Here we developed a technique for improving the detection of hydrophilic QACs using ammonium sulfate (AS) in matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS). In MALDI mass spectrometry for brain homogenates, the addition of AS greatly increased the signal intensities of Car, acetylcarnitine (AcCar), and glycerophosphocholine (GPC) by approximately 300-, 700-, and 2500-fold. The marked improvement required a higher AS concentration than that needed for suppressing the potassium adduction on phosphatidylcholine and 2,5-dihydroxybenzoic acid. Adding AS also increased the signal intensities of Car, AcCar, and GPC by approximately 10-, 20-, and 40-fold in MALDI-IMS. Consequently, the distributions of five hydrophilic QACs (Car, AcCar, GPC, choline, and phosphocholine) were simultaneously visualized by this technique. The distinct mechanism from other techniques such as improved matrix application, derivatization, or postionization suggests the great potential of AS addition to achieve higher sensitivity of MALDI-IMS for various analytes.

  13. Use of matrix assisted laser desorption ionisation-time of flight mass spectrometry in a paediatric clinical laboratory for identification of bacteria commonly isolated from cystic fibrosis patients.

    Desai, Ankita Patel; Stanley, Theresa; Atuan, Maria; McKey, Jonelle; Lipuma, John J; Rogers, Beverly; Jerris, Robert

    2012-09-01

    Matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) has been described as a rapid, accurate method for bacterial identification. To investigate the ability of the technique, using the unamended database supplied with the system, to identify bacteria commonly isolated in cystic fibrosis (CF) patients. Organisms commonly isolated from CF patients identified by MALDI-TOF MS were compared to conventional phenotypic and genotypic analyses. For MALDI-TOF MS, the direct colony technique was used routinely with one extraction procedure performed on a mucoid Pseudomonas aeruginosa. For 24 unique CF specimens, workload comparison and time to identification were assessed. Of 464 tested isolates, conventional (phenotypic and genotypic) identification compared to MALDI-TOF MS showed complete genus, species agreement in 92%, with genus agreement in 98%. This included 29 isolates within the Burkholderia cepacia complex. All 29 were correctly identified to the genus level and 24 of these were speciated. Time to identification with 47 bacterial isolates from 24 CF patients showed identification of 85% of isolates by MALDI-TOF MS at 48 h of incubation, compared to only 34% with conventional methods. Using the unamended database supplied with the system, MALDI-TOF MS provides rapid and reliable identification of bacteria isolated from CF specimens. Time to identification studies showed that the use of same day, same method for organism identification will decrease time to result and optimise microbiology workflow.

  14. Fast and reliable diagnosis of XDR Acinetobacter baumannii meningitis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Brunetti, Grazia; Ceccarelli, Giancarlo; Giordano, Alessandra; Navazio, Anna Sara; Vittozzi, Pietro; Venditti, Mario; Raponi, Giammarco

    2018-01-01

    Bacterial meningitis is a medical emergency needing quick and timely diagnosis. Even though meningitis caused by Acinetobacter baumannii is relatively rare, it is associated with high mortality rates especially in neurosurgery patients and represents a serious therapeutic problem due to the limited penetration of effective antibiotics into the cerebrospinal fluid. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) has been effectively used as a rapid method for microbial identification. In this case report we identified A. baumanni by MALDI-TOF technique directly from the CSF drawn from the external ventricular drainage of a patient with severe confusional state and signs of meningism. Simultaneously the antibiotic susceptibility test was performed by automated method from the pellet of the broth-enriched sample. The MALDI-TOF technique allowed microbial identification in less than 30 minutes, and the susceptibility test result was available in eight hours, thus allowing a fast diagnosis ready for prompt and targeted antimicrobial therapy.

  15. Identification of pathogenic microorganisms directly from positive blood vials by matrix-assisted laser desorption/ionization time of flight mass spectrometry

    Nonnemann, Bettina; Tvede, Michael; Bjarnsholt, Thomas

    2013-01-01

    Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is a promising and fast method for identifying fungi and bacteria directly from positive blood cultures. Various pre-treatment methods for MALDI-TOF MS identification have been reported for this purpose. In......-house results for identification of bacterial colonies by MALDI-TOF MS using a cut-off score of 1.5 did not reduce the diagnostic accuracy compared with the recommended cut-off score of 1.8. A 3-month consecutive study of positive blood cultures was carried out in our laboratory to evaluate whether...... the Sepsityper™ Kit (Bruker Daltonics) with Biotyper 2.0 software could be used as a fast diagnostic tool for bacteria and fungi and whether a 1.5 cut-off score could improve species identification compared with the recommended score of 1.8. Two hundred and fifty-six positive blood vials from 210 patients and 19...

  16. Matrix-assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Can Precisely Discriminate the Lineages of Listeria monocytogenes and Species of Listeria.

    Ojima-Kato, Teruyo; Yamamoto, Naomi; Takahashi, Hajime; Tamura, Hiroto

    2016-01-01

    The genetic lineages of Listeria monocytogenes and other species of the genus Listeria are correlated with pathogenesis in humans. Although matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has become a prevailing tool for rapid and reliable microbial identification, the precise discrimination of Listeria species and lineages remains a crucial issue in clinical settings and for food safety. In this study, we constructed an accurate and reliable MS database to discriminate the lineages of L. monocytogenes and the species of Listeria (L. monocytogenes, L. innocua, L. welshimeri, L. seeligeri, L. ivanovii, L. grayi, and L. rocourtiae) based on the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum (S10-GERMS) proteotyping method, which relies on both genetic information (genomics) and observed MS peaks in MALDI-TOF MS (proteomics). The specific set of eight biomarkers (ribosomal proteins L24, L6, L18, L15, S11, S9, L31 type B, and S16) yielded characteristic MS patterns for the lineages of L. monocytogenes and the different species of Listeria, and led to the construction of a MS database that was successful in discriminating between these organisms in MALDI-TOF MS fingerprinting analysis followed by advanced proteotyping software Strain Solution analysis. We also confirmed the constructed database on the proteotyping software Strain Solution by using 23 Listeria strains collected from natural sources.

  17. Differentiation of clinically relevant Mucorales Rhizopus microsporus and R. arrhizus by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

    Dolatabadi, Somayeh; Kolecka, Anna; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

    2015-07-01

    This study addresses the usefulness of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS for reliable identification of the two most frequently occurring clinical species of Rhizopus, namely Rhizopus arrhizus with its two varieties, arrhizus and delemar, and Rhizopus microsporus. The test-set comprised 38 isolates of clinical and environmental origin previously identified by internal transcribed spacer (ITS) sequencing of rDNA. Multi-locus sequence data targeting three gene markers (ITS, ACT, TEF ) showed two monophylic clades for Rhizopus arrhizus and Rhizopus microsporus (bootstrap values of 99 %). Cluster analysis confirmed the presence of two distinct clades within Rhizopus arrhizus representing its varieties arrhizus and delemar. The MALDI Biotyper 3.0 Microflex LT platform (Bruker Daltonics) was used to confirm the distinction between Rhizopus arrhizus and Rhizopus microsporus and the presence of two varieties within the species Rhizopus arrhizus. An in-house database of 30 reference main spectra (MSPs) was initially tested for correctness using commercially available databases of Bruker Daltonics. By challenging the database with the same strains of which an in-house database was created, automatic identification runs confirmed that MALDI-TOF MS is able to recognize the strains at the variety level. Based on principal component analysis, two MSP dendrograms were created and showed concordance with the multi-locus tree; thus, MALDI-TOF MS is a useful tool for diagnostics of mucoralean species.

  18. A novel cluster of Mycobacterium abscessus complex revealed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS).

    Suzuki, Hiromichi; Yoshida, Shiomi; Yoshida, Atsushi; Okuzumi, Katsuko; Fukusima, Atsuhito; Hishinuma, Akira

    2015-12-01

    Mycobacterium abscessus complex is a rapidly growing mycobacterium consisting of 3 subspecies, M. abscessus, Mycobacterium massiliense, and Mycobacterium bolletii. However, rapid and accurate species identification is difficult. We first evaluated a suitable protocol of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for distinguishing these subspecies. Then, we studied spectral signals by MALDI-TOF MS in 59 M. abscessus, 42 M. massiliense, and 2 M. bolletii. Among several specific spectral signals, 4 signals clearly differentiate M. massiliense from the other 2 subspecies, M. abscessus and M. bolletii. Moreover, 6 of the 42 M. massiliense isolates showed a spectral pattern similar to M. abscessus. These isolates correspond to the distinctive class of M. massiliense (cluster D) which is closer to M. abscessus by the previous variable number tandem repeat analysis. These results indicate that MALDI-TOF MS is not only useful for the identification of 3 subspecies of M. abscessus complex but also capable of distinguishing clusters of M. massiliense. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Stevenson, Lindsay G; Drake, Steven K; Murray, Patrick R

    2010-02-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry is a rapid, accurate method for identifying bacteria and fungi recovered on agar culture media. We report herein a method for the direct identification of bacteria in positive blood culture broths by MALDI-TOF mass spectrometry. A total of 212 positive cultures were examined, representing 32 genera and 60 species or groups. The identification of bacterial isolates by MALDI-TOF mass spectrometry was compared with biochemical testing, and discrepancies were resolved by gene sequencing. No identification (spectral score of blood culture broth. Of the bacteria with a spectral score of > or = 1.7, 162 (95.3%) of 170 isolates were correctly identified. All 8 isolates of Streptococcus mitis were misidentified as being Streptococcus pneumoniae isolates. This method provides a rapid, accurate, definitive identification of bacteria within 1 h of detection in positive blood cultures with the caveat that the identification of S. pneumoniae would have to be confirmed by an alternative test.

  20. Instant screening and verification of carbapenemase activity in Bacteroides fragilis in positive blood culture, using matrix-assisted laser desorption ionization--time of flight mass spectrometry.

    Johansson, Åsa; Nagy, Elisabeth; Sóki, József

    2014-08-01

    Rapid identification of isolates in positive blood cultures are of great importance to secure correct treatment of septicaemic patients. As antimicrobial resistance is increasing, rapid detection of resistance is crucial. Carbapenem resistance in Bacteroides fragilis associated with cfiA-encoded class B metallo-beta-lactamase is emerging. In our study we spiked blood culture bottles with 26 B. fragilis strains with various cfiA-status and ertapenem MICs. By using main spectra specific for cfiA-positive and cfiA-negative B. fragilis strains, isolates could be screened for resistance. To verify strains that were positive in the screening, a carbapenemase assay was performed where the specific peaks of intact and hydrolysed ertapenem were analysed with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). We show here that it is possible to correctly identify B. fragilis and to screen for enzymic carbapenem resistance directly from the pellet of positive blood cultures. The carbapenemase assay to verify the presence of the enzyme was successfully performed on the pellet from the direct identification despite the presence of blood components. The result of the procedure was achieved in 3 h. Also the Bruker mass spectrometric β-lactamase assay (MSBL assay) prototype software was proven not only to be based on an algorithm that correlated with the manual inspection of the spectra, but also to improve the interpretation by showing the variation in the dataset. © 2014 The Authors.

  1. Rapid identification of bacteria in positive blood culture by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Schmidt, V; Jarosch, A; März, P; Sander, C; Vacata, V; Kalka-Moll, W

    2012-03-01

    Blood culture is probably the most significant specimen used for the diagnosis of bacterial infections, especially for bloodstream infections. In the present study, we compared the resin-containing BD BACTEC™ Plus-Aerobic (Becton Dickinson), non-charcoal-containing BacT/Alert(®) SA (bioMérieux), and charcoal-containing BacT/Alert(®) FA (bioMérieux) blood culture bottles with direct identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). A total of 103 bacterial isolates, from clinical blood cultures, representing the most frequent 13 genera and 24 species were examined. Bacteria were extracted from positive blood culture broth by density centrifugation and then subjected to identification by MALDI-TOF MS using two different volumes and chemical treatments. Overall, correct identification by MALDI-TOF MS was obtained for the BD BACTEC™ Plus-Aerobic, BacT/Alert(®) SA, and BacT/Alert(®) FA blood culture bottles in 72%, 45.6%, and 23%, respectively, for gram-negative bacteria in 86.6%, 69.2%, and 47.1%, respectively, and for gram-positive bacteria in 60.0%, 28.8%, and 5.4%, respectively. The lack of identification was observed mainly with viridans streptococci. Depending on the blood culture bottles used in routine diagnostic procedures and the protocol for bacterial preparation, the applied MALDI-TOF MS represents an efficient and rapid method for direct bacterial identification.

  2. Rapid identification of bacteria from positive blood culture bottles by use of matrix-assisted laser desorption-ionization time of flight mass spectrometry fingerprinting.

    Christner, Martin; Rohde, Holger; Wolters, Manuel; Sobottka, Ingo; Wegscheider, Karl; Aepfelbacher, Martin

    2010-05-01

    Early and adequate antimicrobial therapy has been shown to improve the clinical outcome in bloodstream infections (BSI). To provide rapid pathogen identification for targeted treatment, we applied matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry fingerprinting to bacteria directly recovered from blood culture bottles. A total of 304 aerobic and anaerobic blood cultures, reported positive by a Bactec 9240 system, were subjected in parallel to differential centrifugation with subsequent mass spectrometry fingerprinting and reference identification using established microbiological methods. A representative spectrum of bloodstream pathogens was recovered from 277 samples that grew a single bacterial isolate. Species identification by direct mass spectrometry fingerprinting matched reference identification in 95% of these samples and worked equally well for aerobic and anaerobic culture bottles. Application of commonly used score cutoffs to classify the fingerprinting results led to an identification rate of 87%. Mismatching mostly resulted from insufficient bacterial numbers and preferentially occurred with Gram-positive samples. The respective spectra showed low concordance to database references and were effectively rejected by score thresholds. Spiking experiments and examination of the respective study samples even suggested applicability of the method to mixed cultures. With turnaround times around 100 min, the approach allowed for reliable pathogen identification at the day of blood culture positivity, providing treatment-relevant information within the critical phase of septic illness.

  3. A simple method for rapid microbial identification from positive monomicrobial blood culture bottles through matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Lin, Jung-Fu; Ge, Mao-Cheng; Liu, Tsui-Ping; Chang, Shih-Cheng; Lu, Jang-Jih

    2017-06-30

    Rapid identification of microbes in the bloodstream is crucial in managing septicemia because of its high disease severity, and direct identification from positive blood culture bottles through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can shorten the turnaround time. Therefore, we developed a simple method for rapid microbiological identification from positive blood cultures by using MALDI-TOF MS. We modified previously developed methods to propose a faster, simpler and more economical method, which includes centrifugation and hemolysis. Specifically, our method comprises two-stage centrifugation with gravitational acceleration (g) at 600g and 3000g, followed by the addition of a lysis buffer and another 3000g centrifugation. In total, 324 monomicrobial bacterial cultures were identified. The success rate of species identification was 81.8%, which is comparable with other complex methods. The identification success rate was the highest for Gram-negative aerobes (85%), followed by Gram-positive aerobes (78.2%) and anaerobes (67%). The proposed method requires less than 10 min, costs less than US$0.2 per usage, and facilitates batch processing. We conclude that this method is feasible for clinical use in microbiology laboratories, and can serve as a reference for treatments or further complementary diagnostic testing. Copyright © 2017. Published by Elsevier B.V.

  4. Direct identification of bacteria from positive BacT/ALERT blood culture bottles using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

    Mestas, Javier; Felsenstein, Susanna; Bard, Jennifer Dien

    2014-11-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is a fast and robust method for the identification of bacteria. In this study, we evaluate the performance of a laboratory-developed lysis method (LDT) for the rapid identification of bacteria from positive BacT/ALERT blood culture bottles. Of the 168 positive bottles tested, 159 were monomicrobial, the majority of which were Gram-positive organisms (61.0% versus 39.0%). Using a cut-off score of ≥1.7, 80.4% of the organisms were correctly identified to the species level, and the identification rate of Gram-negative organisms (90.3%) was found to be significantly greater than that of Gram-positive organisms (78.4%). The simplicity and cost-effectiveness of the LDT enable it to be fully integrated into the routine workflow of the clinical microbiology laboratory, allowing for rapid identification of Gram-positive and Gram-negative bacteria within an hour of blood culture positivity. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Direct identification of bacteria in blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a new methodological approach.

    Kroumova, Vesselina; Gobbato, Elisa; Basso, Elisa; Mucedola, Luca; Giani, Tommaso; Fortina, Giacomo

    2011-08-15

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has recently been demonstrated to be a powerful tool for the rapid identification of bacteria from growing colonies. In order to speed up the identification of bacteria, several authors have evaluated the usefulness of this MALDI-TOF MS technology for the direct and quick identification bacteria from positive blood cultures. The results obtained so far have been encouraging but have also shown some limitations, mainly related to the bacterial growth and to the presence of interference substances belonging to the blood cultures. In this paper, we present a new methodological approach that we have developed to overcome these limitations, based mainly on an enrichment of the sample into a growing medium before the extraction process, prior to mass spectrometric analysis. The proposed method shows important advantages for the identification of bacterial strains, yielding an increased identification score, which gives higher confidence in the results. Copyright © 2011 John Wiley & Sons, Ltd.

  6. Effects of solid-medium type on routine identification of bacterial isolates by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Anderson, Neil W; Buchan, Blake W; Riebe, Katherine M; Parsons, Lauren N; Gnacinski, Stacy; Ledeboer, Nathan A

    2012-03-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a rapid method for the identification of bacteria. Factors that may alter protein profiles, including growth conditions and presence of exogenous substances, could hinder identification. Bacterial isolates identified by conventional methods were grown on various media and identified using the MALDI Biotyper (Bruker Daltonics, Billerica, MA) using a direct smear method and an acid extraction method. Specimens included 23 Pseudomonas isolates grown on blood agar, Pseudocel (CET), and MacConkey agar (MAC); 20 Staphylococcus isolates grown on blood agar, colistin-nalidixic acid agar (CNA), and mannitol salt agar (MSA); and 25 enteric isolates grown on blood agar, xylose lysine deoxycholate agar (XLD), Hektoen enteric agar (HE), salmonella-shigella agar (SS), and MAC. For Pseudomonas spp., the identification rate to genus using the direct method was 83% from blood, 78% from MAC, and 94% from CET. For Staphylococcus isolates, the identification rate to genus using the direct method was 95% from blood, 75% from CNA, and 95% from MSA. For enteric isolates, the identification rate to genus using the direct method was 100% from blood, 100% from MAC, 100% from XLD, 92% from HE, and 87% from SS. Extraction enhanced identification rates. The direct method of MALDI-TOF analysis of bacteria from selective and differential media yields identifications of varied confidence. Notably, Staphylococci spp. from CNA exhibit low identification rates. Extraction enhances identification rates and is recommended for colonies from this medium.

  7. Applications of copolymer for rapid identification of bacteria in blood culture broths using matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Ashizawa, Kazuho; Murata, Syota; Terada, Takashi; Ito, Daisuke; Bunya, Masaru; Watanabe, Koji; Teruuchi, Yoko; Tsuchida, Sachio; Satoh, Mamoru; Nishimura, Motoi; Matsushita, Kazuyuki; Sugama, Yuji; Nomura, Fumio

    2017-08-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can be used to identify pathogens in blood culture samples. However, sample pretreatment is needed for direct identification of microbes in blood culture bottles. Conventional protocols are complex and time-consuming. Therefore, in this study, we developed a method for collecting bacteria using polyallylamine-polystyrene copolymer for application in wastewater treatment technology. Using representative bacterial species Escherichia coli and Staphylococcus capitis, we found that polyallylamine-polystyrene can form visible aggregates with bacteria, which can be identified using MALDI-TOF MS. The processing time of our protocol was as short as 15min. Hemoglobin interference in MALDI spectra analysis was significantly decreased in our method compared with the conventional method. In a preliminary experiment, we evaluated the use of our protocol to identify clinical isolates from blood culture bottles. MALDI-TOF MS-based identification of 17 strains from five bacterial species (E. coli, Klebsiella pneumoniae, Enterococcus faecalis, S. aureus, and S. capitis) collected by our protocol was satisfactory. Prospective large-scale studies are needed to further evaluate the clinical application of this novel and simple method of collecting bacteria in blood culture bottles. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

    Jeong, Youngsu; Lee, Jonghee; Kim, Seongsoo [Agency for Defense Development, Daejeon (Korea, Republic of)

    2013-09-15

    The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

  9. Atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry of friction modifier additives analyzed directly from base oil solutions.

    Widder, Lukas; Brennerb, Josef; Huttera, Herbert

    2014-01-01

    To develop new products and to apply measures of quality control quick and simple accessibility of additive composition in automo- tive lubrication is important. The aim of this study was to investigate the possibility of analyzing organic friction modifier additives by means of atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry [AP-MALDI-MS] from lubricant solu- tions without the use of additional separation techniques. Analyses of selected friction modifier ethoxylated tallow amines and oleic acid amide were compared using two ionization methods, positive-ion electrospray ionization (ESI) and AP-MALDI, using a LTQ Orbitrap mass spectrometer. Pure additives were characterized from solvent solutions, as well as from synthetic and mineral base oil mixtures. Detected ions of pure additive samples consisted mainly of [M + H]+, but also alkaLi metal adducts [M + Na]+ and [M + K]+ could be seen. Characterizations of blends of both friction modifiers from the base oil mixtures were carried out as well and showed significant inten- sities for several additive peaks. Thus, this work shows a method to directly analyze friction modifier additives used in the automotive industry from an oil blend via the use of AP-MALDI without any further separation steps. The method presented will further simplify the acquisition of data on lubricant composition and additives. Furthermore, it allows the perspective of analyzing additive reaction products directly from formulated oil blends.

  10. Evaluation of VITEK mass spectrometry (MS), a matrix-assisted laser desorption ionization time-of-flight MS system for identification of anaerobic bacteria.

    Lee, Wonmok; Kim, Myungsook; Yong, Dongeun; Jeong, Seok Hoon; Lee, Kyungwon; Chong, Yunsop

    2015-01-01

    By conventional methods, the identification of anaerobic bacteria is more time consuming and requires more expertise than the identification of aerobic bacteria. Although the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems are relatively less studied, they have been reported to be a promising method for the identification of anaerobes. We evaluated the performance of the VITEK MS in vitro diagnostic (IVD; 1.1 database; bioMérieux, France) in the identification of anaerobes. We used 274 anaerobic bacteria isolated from various clinical specimens. The results for the identification of the bacteria by VITEK MS were compared to those obtained by phenotypic methods and 16S rRNA gene sequencing. Among the 249 isolates included in the IVD database, the VITEK MS correctly identified 209 (83.9%) isolates to the species level and an additional 18 (7.2%) at the genus level. In particular, the VITEK MS correctly identified clinically relevant and frequently isolated anaerobic bacteria to the species level. The remaining 22 isolates (8.8%) were either not identified or misidentified. The VITEK MS could not identify the 25 isolates absent from the IVD database to the species level. The VITEK MS showed reliable identifications for clinically relevant anaerobic bacteria.

  11. Hydrophilic interaction chromatography coupled matrix assisted laser desorption/ionization mass spectrometry for molecular analysis of organic compounds in medicines, tea, and coffee

    Wang, Renqi

    2013-01-01

    Natural occurring organic compounds from food, natural organic matter, as well as metabolic products have received intense attention in current chemical and biological studies. Examination of unknown compounds in complex sample matrices is hampered by the limited choices for data readout and molecular elucidation. Herein, we report a generic method of hydrophilic interaction chromatography (HILIC) coupled with matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) for the rapid characterization of ingredients in pharmaceutical compounds, tea, and coffee. The analytes were first fractionated using a cationic HILIC column prior to MALDI-MS analyses. It was found that the retention times of a compound arising from different samples were consistent under the same conditions. Accordingly, molecules can be readily characterized by both the mass and chromatographic retention time. The retention behaviors of acidic and basic compounds on the cationic HILIC column were found to be significantly influenced by the pH of mobile phases, whereas neutral compounds depicted a constant retention time at different pH. The general HILIC-MALDI-MS method is feasible for fast screening of naturally occurring organic compounds. A series of homologs can be determined if they have the same retention behavior. Their structural features can be elucidated by considering their mass differences and hydrophilic properties as determined by HILIC chromatogram. © 2013 The Royal Society of Chemistry.

  12. Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry rapid detection of carbapenamase activity in Acinetobacter baumannii isolates

    Noha Abouseada

    2017-01-01

    Full Text Available Introduction: Carbapenamase-producing Acinetobacter baumannii are an increasing threat in hospitals and Intensive Care Units. Accurate and rapid detection of carbapenamase producers has a great impact on patient improvement and aids in implementation of infection control measures. Aim: In this study, we describe the use of matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI TOF MS to identify carbapenamase-producing A. baumannii isolates in up to 3 h. Isolates and Methods: A total of 50 A. baumannii isolates (of which 39 were carabapenamase producers were tested using MALDI TOF MS. Isolates were incubated for 3 h with 0.25 mg/ml up to 2 mg/ml of imipenem (IMP at 37°C. Supernatants were analysed by MALDI TOF to analyse peaks corresponding to IMP (300 Da and an IMP metabolite (254 Da using UltrafleXtreme (Bruker Daltonics, Bremen, Germany. Results: All carbapenamase-producing isolates were evidenced by the disappearance or reduction in intensity of the 300 Da peak of IPM and the appearance of a 254 Da peak of the IPM metabolite. In isolates that did not produce carbapenamase, the IPM 300 Da peak remained intact. Conclusion: MALDI TOF is a promising tool in the field of diagnostic microbiology that has the ability to transfer identification and antimicrobial susceptibility testing time from days to hours.

  13. Applications of interface controlled pulsed-laser deposited polymer films in field-effect transistors

    Adil, Danish; Ukah, Ndubuisi; Guha, Suchi; Gupta, Ram; Ghosh, Kartik

    2010-03-01

    Matrix assisted pulsed laser evaporation, a derivative of pulsed laser deposition (PLD), is an alternative method of depositing polymer and biomaterial films that allows homogeneous film coverage of high molecular weight organic materials for layer-by-layer growth without any laser induced damage. Polyfluorene (PF)-based conjugated polymers have attracted considerable attention in organic field-effect transistors (FETs). A co-polymer of PF (PFB) was deposited as a thin film using matrix assisted PLD employing a KrF excimer laser. Electrical characteristics of FETs fabricated using these PLD grown films were compared to those of FETs using spin-coated films. We show that threshold voltages, on/off ratios, and charge carrier motilities are significantly improved in PLD grown films. This is attributed to an improved dielectric-polymer interface.

  14. Distribution of erlotinib in rash and normal skin in cancer patients receiving erlotinib visualized by matrix assisted laser desorption/ionization mass spectrometry imaging.

    Nishimura, Meiko; Hayashi, Mitsuhiro; Mizutani, Yu; Takenaka, Kei; Imamura, Yoshinori; Chayahara, Naoko; Toyoda, Masanori; Kiyota, Naomi; Mukohara, Toru; Aikawa, Hiroaki; Fujiwara, Yasuhiro; Hamada, Akinobu; Minami, Hironobu

    2018-04-06

    The development of skin rashes is the most common adverse event observed in cancer patients treated with epidermal growth factor receptor-tyrosine kinase inhibitors such as erlotinib. However, the pharmacological evidence has not been fully revealed. Erlotinib distribution in the rashes was more heterogeneous than that in the normal skin, and the rashes contained statistically higher concentrations of erlotinib than adjacent normal skin in the superficial skin layer (229 ± 192 vs. 120 ± 103 ions/mm 2 ; P = 0.009 in paired t -test). LC-MS/MS confirmed that the concentration of erlotinib in the skin rashes was higher than that in normal skin in the superficial skin layer (1946 ± 1258 vs. 1174 ± 662 ng/cm 3 ; P = 0.028 in paired t -test). The results of MALDI-MSI and LC-MS/MS were well correlated (coefficient of correlation 0.879, P distribution of erlotinib in the skin tissue was visualized using non-labeled MALDI-MSI. Erlotinib concentration in the superficial layer of the skin rashes was higher than that in the adjacent normal skin. We examined patients with advanced pancreatic cancer who developed skin rashes after treatment with erlotinib and gemcitabine. We biopsied both the rash and adjacent normal skin tissues, and visualized and compared the distribution of erlotinib within the skin using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). The tissue concentration of erlotinib was also measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with laser microdissection.

  15. High energy HF pulsed lasers

    Patterson, E.L.; Gerber, R.A.

    1976-01-01

    Recent experiments show that pulsed HF lasers are capable of producing high energy with good efficiency. Preliminary experiments show that the laser radiation from the high-gain medium can be controlled with a low-power probe laser beam or with low-level feedback. These results indicate that the HF laser may have potential for second-generation laser fusion experiments

  16. Emerging and Future Applications of Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry in the Clinical Microbiology Laboratory: A Report of the Association for Molecular Pathology.

    Doern, Christopher D; Butler-Wu, Susan M

    2016-11-01

    The performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) for routine bacterial and yeast identification as well as direct-from-blood culture bottle identification has been thoroughly evaluated in the peer-reviewed literature. Microbiologists are now moving beyond these methods to apply MS to other areas of the diagnostic process. This review discusses the emergence of advanced matrix-assisted laser desorption ionization time-of-flight MS applications, including the identification of filamentous fungi and mycobacteria and the current and future state of antimicrobial resistance testing. Copyright © 2016 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  17. Capillary isoelectric focusing of probiotic bacteria from cow's milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification

    Horká, Marie; Karásek, Pavel; Šalplachta, Jiří; Růžička, F.; Vykydalová, Marie; Kubesová, Anna; Dráb, V.; Roth, Michal; Šlais, Karel

    2013-01-01

    Roč. 788, JUL (2013), s. 193-199 ISSN 0003-2670 R&D Projects: GA ČR(CZ) GAP106/12/0522; GA MV VG20102015023 Institutional support: RVO:68081715 Keywords : capillary isoelectric focusing * matrix-assisted laser desorption/ionization time-of-flight mass spectrometry * lactic acid bacteria Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.517, year: 2013

  18. Novel, Improved Sample Preparation for Rapid, Direct Identification from Positive Blood Cultures Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry

    Schubert, Sören; Weinert, Kirsten; Wagner, Chris; Gunzl, Beatrix; Wieser, Andreas; Maier, Thomas; Kostrzewa, Markus

    2011-01-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used for rapid and reliable identification of bacteria and yeast grown on agar plates. Moreover, MALDI-TOF MS also holds promise for bacterial identification from blood culture (BC) broths in hospital laboratories. The most important technical step for the identification of bacteria from positive BCs by MALDI-TOF MS is sample preparation to remove blood cells and host proteins. We present a m...

  19. Identification of Blood Culture Isolates Directly from Positive Blood Cultures by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and a Commercial Extraction System: Analysis of Performance, Cost, and Turnaround Time

    Lagacé-Wiens, Philippe R. S.; Adam, Heather J.; Karlowsky, James A.; Nichol, Kimberly A.; Pang, Paulette F.; Guenther, Jodi; Webb, Amanda A.; Miller, Crystal; Alfa, Michelle J.

    2012-01-01

    Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry represents a revolution in the rapid identification of bacterial and fungal pathogens in the clinical microbiology laboratory. Recently, MALDI-TOF has been applied directly to positive blood culture bottles for the rapid identification of pathogens, leading to reductions in turnaround time and potentially beneficial patient impacts. The development of a commercially available extraction kit (Bruker Sepsit...

  20. An in-house assay is superior to Sepsityper for direct matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification of yeast species in blood cultures.

    Bidart, Marie; Bonnet, Isabelle; Hennebique, Aurélie; Kherraf, Zine Eddine; Pelloux, Hervé; Berger, François; Cornet, Muriel; Bailly, Sébastien; Maubon, Danièle

    2015-05-01

    We developed an in-house assay for the direct identification, by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, of yeasts in blood culture. Sixty-one representative strains from 12 species were analyzed in spiked blood cultures. Our assay accurately identified 95 of 107 (88.8%) positive blood cultures and outperformed the commercial Sepsityper kit (81.7% identification). Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  1. Coumarins as new matrices for matrix-assisted laser-desorption/ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of hydrophobic compounds

    Wang, Hang, E-mail: hangwang@sjtu.edu.cn [Instrumental Analysis Center, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240 (China); Dai, Bona [Instrumental Analysis Center, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240 (China); Liu, Bin [Key Laboratory of Kidney Disease Pathogenesis and Intervention of Hubei Province, College of Medicine, Hubei Polytechnic University, Huangshi, Hubei 435003 (China); Lu, Han [Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine (SJTU-SM), 197, Rui Jin Er Road, Shanghai 200025 (China)

    2015-07-02

    Highlights: • Coumarins were used as new MALDI matrices. • Coumarins were used for MALDI-FT ICR MS detection of hydrophobic compounds. • DCA had improvement in detection sensitivity, stability, selectivity and reproducibility. • DCA was applied to sterols detection in yeast cells. - Abstract: Hydrophobic compounds with hydroxyl, aldehyde or ketone groups are generally difficult to detect using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), because these compounds have low proton affinity and are poorly ionized by MALDI. Herein, coumarins have been used as new matrices for MALDI-MS analysis of a variety of hydrophobic compounds with low ionization efficiency, including steroids, coenzyme Q10, a cyclic lipopeptide and cholesterol oleate. Five coumarins, including coumarin, umbelliferone, esculetin, 7-hydroxycoumarin-3-carboxylic acid (HCA) and 6,7-dihydroxycoumarin-3-carboxylic acid (DCA), were compared with the conventional matrices of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA). Coumarins with hydroxyl or carboxylic acid groups enabled detection. Taking DCA as an example, this matrix proved to be superior to DHB or CHCA in detection sensitivity, stability, spot-to-spot and sample-to-sample reproducibility, and accuracy. DCA increased the stability of the target compounds and decreased the loss of water. The [M + Na]{sup +} peaks were observed for all target compounds by adding NaCl as an additive, and the [M − H{sub 2}O + H]{sup +} and [M + H]{sup +} peaks decreased. DCA was selected for the identification of sterols in yeast cells, and thirteen sterols were detected by Fourier transform ion cyclotron resonance (FT ICR) mass spectrometry. This work demonstrates the potential of DCA as a new matrix for detection of hydrophobic molecules by MALDI-MS and provides an alternative tool for screening sterols in antifungal research.

  2. The Exploration of Peptide Biomarkers in Malignant Pleural Effusion of Lung Cancer Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

    Jing Xu

    2017-01-01

    Full Text Available Background. Diagnoses of malignant pleural effusion (MPE are a crucial problem in clinics. In our study, we compared the peptide profiles of MPE and tuberculosis pleural effusion (TPE to investigate the value of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS in diagnosis of MPE. Material and Methods. The 46 MPE and 32 TPE were randomly assigned to training set and validation set. Peptides were isolated by weak cation exchange magnetic beads and peaks in the m/z range of 800–10000 Da were analyzed. Comparing the peptide profile between 30 MPE and 22 TPE samples in training set by ClinProTools software, we screened the specific biomarkers and established a MALDI-TOF-MS classification of MPE. Finally, the other 16 MPE and 10 TPE were included to verify the model. We additionally determined carcinoembryonic antigen (CEA in MPE and TPE samples using electrochemiluminescent immunoassay method. Results. Five peptide peaks (917.37 Da, 4469.39 Da, 1466.5 Da, 4585.21 Da, and 3216.87 Da were selected to separate MPE and TPE by MALDI-TOF-MS. The sensitivity, specificity, and accuracy of the classification were 93.75%, 100%, and 96.15%, respectively, after blinded test. The sensitivity of CEA was significantly lower than MALDI-TOF-MS classification (P=0.035. Conclusions. The results indicate MALDI-TOF-MS is a potential method for diagnosing MPE.

  3. Use of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of molds of the Fusarium genus.

    Triest, David; Stubbe, Dirk; De Cremer, Koen; Piérard, Denis; Normand, Anne-Cécile; Piarroux, Renaud; Detandt, Monique; Hendrickx, Marijke

    2015-02-01

    The rates of infection with Fusarium molds are increasing, and a diverse number of Fusarium spp. belonging to different species complexes can cause infection. Conventional species identification in the clinical laboratory is time-consuming and prone to errors. We therefore evaluated whether matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a useful alternative. The 289 Fusarium strains from the Belgian Coordinated Collections of Microorganisms (BCCM)/Institute of Hygiene and Epidemiology Mycology (IHEM) culture collection with validated sequence-based identities and comprising 40 species were used in this study. An identification strategy was developed, applying a standardized MALDI-TOF MS assay and an in-house reference spectrum database. In vitro antifungal testing was performed to assess important differences in susceptibility between clinically relevant species/species complexes. We observed that no incorrect species complex identifications were made by MALDI-TOF MS, and 82.8% of the identifications were correct to the species level. This success rate was increased to 91% by lowering the cutoff for identification. Although the identification of the correct species complex member was not always guaranteed, antifungal susceptibility testing showed that discriminating between Fusarium species complexes can be important for treatment but is not necessarily required between members of a species complex. With this perspective, some Fusarium species complexes with closely related members can be considered as a whole, increasing the success rate of correct identifications to 97%. The application of our user-friendly MALDI-TOF MS identification approach resulted in a dramatic improvement in both time and accuracy compared to identification with the conventional method. A proof of principle of our MALDI-TOF MS approach in the clinical setting using recently isolated Fusarium strains demonstrated its validity. Copyright © 2015

  4. Independent assessment of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) sample preparation quality: A novel statistical approach for quality scoring.

    Kooijman, Pieter C; Kok, Sander J; Weusten, Jos J A M; Honing, Maarten

    2016-05-05

    Preparation of samples according to an optimized method is crucial for accurate determination of polymer sample characteristics by Matrix-Assisted Laser Desorption Ionization (MALDI) analysis. Sample preparation conditions such as matrix choice, cationization agent, deposition technique or even the deposition volume should be chosen to suit the sample of interest. Many sample preparation protocols have been developed and employed, yet finding the optimal sample preparation protocol remains a challenge. Because an objective comparison between the results of diverse protocols is not possible, "gut-feeling" or "good enough" is often decisive in the search for an optimum. This implies that sub-optimal protocols are used, leading to a loss of mass spectral information quality. To address this problem a novel analytical strategy based on MALDI imaging and statistical data processing was developed in which eight parameters were formulated to objectively quantify the quality of sample deposition and optimal MALDI matrix composition and finally sum up to an overall quality score of the sample deposition. These parameters can be established in a fully automated way using commercially available mass spectrometry imaging instruments without any hardware adjustments. With the newly developed analytical strategy the highest quality MALDI spots were selected, resulting in more reproducible and more valuable spectra for PEG in a variety of matrices. Moreover, our method enables an objective comparison of sample preparation protocols for any analyte and opens up new fields of investigation by presenting MALDI performance data in a clear and concise way. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Identification of rare pathogenic bacteria in a clinical microbiology laboratory: impact of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Seng, Piseth; Abat, Cedric; Rolain, Jean Marc; Colson, Philippe; Lagier, Jean-Christophe; Gouriet, Frédérique; Fournier, Pierre Edouard; Drancourt, Michel; La Scola, Bernard; Raoult, Didier

    2013-07-01

    During the past 5 years, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has become a powerful tool for routine identification in many clinical laboratories. We analyzed our 11-year experience in routine identification of clinical isolates (40 months using MALDI-TOF MS and 91 months using conventional phenotypic identification [CPI]). Among the 286,842 clonal isolates, 284,899 isolates of 459 species were identified. The remaining 1,951 isolates were misidentified and required confirmation using a second phenotypic identification for 670 isolates and using a molecular technique for 1,273 isolates of 339 species. MALDI-TOF MS annually identified 112 species, i.e., 36 species/10,000 isolates, compared to 44 species, i.e., 19 species/10,000 isolates, for CPI. Only 50 isolates required second phenotypic identifications during the MALDI-TOF MS period (i.e., 4.5 reidentifications/10,000 isolates) compared with 620 isolates during the CPI period (i.e., 35.2/10,000 isolates). We identified 128 bacterial species rarely reported as human pathogens, including 48 using phenotypic techniques (22 using CPI and 37 using MALDI-TOF MS). Another 75 rare species were identified using molecular methods. MALDI-TOF MS reduced the time required for identification by 55-fold and 169-fold and the cost by 5-fold and 96-fold compared with CPI and gene sequencing, respectively. MALDI-TOF MS was a powerful tool not only for routine bacterial identification but also for identification of rare bacterial species implicated in human infectious diseases. The ability to rapidly identify bacterial species rarely described as pathogens in specific clinical specimens will help us to study the clinical burden resulting from the emergence of these species as human pathogens, and MALDI-TOF MS may be considered an alternative to molecular methods in clinical laboratories.

  6. Speciation of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry†

    Mandrell, Robert E.; Harden, Leslie A.; Bates, Anna; Miller, William G.; Haddon, William F.; Fagerquist, Clifton K.

    2005-01-01

    Multiple strains of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis isolated from animal, clinical, or food samples have been analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Whole bacterial cells were harvested from colonies or confluent growth on agar and transferred directly into solvent and then to a spot of dried 3-methoxy-4-hydroxycinnamic acid (matrix). Multiple ions in the 5,000- to 15,000-Da mass range were evident in spectra for each strain; one or two ions in the 9,500- to 11,000-Da range were consistently high intensity. “Species-identifying” biomarker ions (SIBIs) were evident from analyses of multiple reference strains for each of the six species, including the genome strains C. jejuni NCTC 11168 and C. jejuni RM1221. Strains grown on nine different combinations of media and atmospheres yielded SIBI masses within ±5 Da with external instrument calibration. The highest-intensity C. jejuni SIBIs were cytosolic proteins, including GroES, HU/HCj, and RplL. Multiple intraspecies SIBIs, corresponding probably to nonsynonymous nucleotide polymorphisms, also provided some intraspecies strain differentiation. MALDI-TOF MS analysis of 75 additional Campylobacter strains isolated from humans, poultry, swine, dogs, and cats revealed (i) associations of SIBI type with source, (ii) strains previously speciated incorrectly, and (iii) “strains” composed of more than one species. MALDI-TOF MS provides an accurate, sensitive, and rapid method for identification of multiple Campylobacter species relevant to public health and food safety. PMID:16204551

  7. Speciation of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Mandrell, Robert E; Harden, Leslie A; Bates, Anna; Miller, William G; Haddon, William F; Fagerquist, Clifton K

    2005-10-01

    Multiple strains of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis isolated from animal, clinical, or food samples have been analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Whole bacterial cells were harvested from colonies or confluent growth on agar and transferred directly into solvent and then to a spot of dried 3-methoxy-4-hydroxycinnamic acid (matrix). Multiple ions in the 5,000- to 15,000-Da mass range were evident in spectra for each strain; one or two ions in the 9,500- to 11,000-Da range were consistently high intensity. "Species-identifying" biomarker ions (SIBIs) were evident from analyses of multiple reference strains for each of the six species, including the genome strains C. jejuni NCTC 11168 and C. jejuni RM1221. Strains grown on nine different combinations of media and atmospheres yielded SIBI masses within +/-5 Da with external instrument calibration. The highest-intensity C. jejuni SIBIs were cytosolic proteins, including GroES, HU/HCj, and RplL. Multiple intraspecies SIBIs, corresponding probably to nonsynonymous nucleotide polymorphisms, also provided some intraspecies strain differentiation. MALDI-TOF MS analysis of 75 additional Campylobacter strains isolated from humans, poultry, swine, dogs, and cats revealed (i) associations of SIBI type with source, (ii) strains previously speciated incorrectly, and (iii) "strains" composed of more than one species. MALDI-TOF MS provides an accurate, sensitive, and rapid method for identification of multiple Campylobacter species relevant to public health and food safety.

  8. High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories.

    van Veen, S Q; Claas, E C J; Kuijper, Ed J

    2010-03-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.

  9. Quantification of the PR-39 cathelicidin compound in porcine blood by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Smolira, Anna; Hałas, Stanisław; Wessely-Szponder, Joanna

    2015-10-15

    The PR-39 porcine cathelicidin occurs naturally in animal neutrophils. Its main function is antimicrobial activity, which potentially can be used in antibiotic treatments in veterinary medicine. Investigations concerning such a use require the detection and quantification of PR-39 in a given sample. The aim of this work is to determine the concentration of PR-39 contained in porcine blood. Prior to matrix-assisted laser desorption/ionization (MALDI) analysis, the porcine blood sample was subjected to crude extraction in order to release the active form of PR-39 from the neutrophil granules. Next, gel filtration chromatography was performed to separate PR-39 from other cathelicidins present in porcine blood. Positive ion MALDI time-of-flight (TOF) mass spectra of the resulting portion of lyophilisate with unknown PR-39 content were acquired in linear mode. To quantify PR-39 in the lyophilisate sample, the standard addition method was applied. The PR-39 concentration obtained in the lyophilisate sample was then converted into the peptide concentration in porcine blood. The linear fit function of the constructed calibration curve indicates an excellent correlation between the PR-39 peak intensity and the added quantity of synthetic PR-39 (R(2) = 0.994) and a low relative standard deviation of the slope = 1.98%. From the x-intercept of the straight line, we estimated the PR-39 concentration in porcine blood to be 20.5 ± 4.6 ng/mL. The MALDI method was successfully applied for the quantitative analysis of PR-39 found in porcine blood. Compared with other available methods, it is relatively easy, inexpensive and not time-consuming. Despite the method having lower accuracy than the enzyme-linked immunosorbent assay (ELISA), the results obtained here, by a much simpler method, are in good agreement with the literature data. Copyright © 2015 John Wiley & Sons, Ltd.

  10. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry for fast and accurate identification of clinically relevant Aspergillus species.

    Alanio, A; Beretti, J-L; Dauphin, B; Mellado, E; Quesne, G; Lacroix, C; Amara, A; Berche, P; Nassif, X; Bougnoux, M-E

    2011-05-01

    New Aspergillus species have recently been described with the use of multilocus sequencing in refractory cases of invasive aspergillosis. The classical phenotypic identification methods routinely used in clinical laboratories failed to identify them adequately. Some of these Aspergillus species have specific patterns of susceptibility to antifungal agents, and misidentification may lead to inappropriate therapy. We developed a matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS)-based strategy to adequately identify Aspergillus species to the species level. A database including the reference spectra of 28 clinically relevant species from seven Aspergillus sections (five common and 23 unusual species) was engineered. The profiles of young and mature colonies were analysed for each reference strain, and species-specific spectral fingerprints were identified. The performance of the database was then tested on 124 clinical and 16 environmental isolates previously characterized by partial sequencing of the β-tubulin and calmodulin genes. One hundred and thirty-eight isolates of 140 (98.6%) were correctly identified. Two atypical isolates could not be identified, but no isolate was misidentified (specificity: 100%). The database, including species-specific spectral fingerprints of young and mature colonies of the reference strains, allowed identification regardless of the maturity of the clinical isolate. These results indicate that MALDI-TOF MS is a powerful tool for rapid and accurate identification of both common and unusual species of Aspergillus. It can give better results than morphological identification in clinical laboratories. © 2010 The Authors. Clinical Microbiology and Infection © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  11. Prediction of Streptococcus uberis clinical mastitis risk using Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) in dairy herds.

    Archer, Simon C; Bradley, Andrew J; Cooper, Selin; Davies, Peers L; Green, Martin J

    2017-09-01

    The purpose of this study was to evaluate whether the risk of Streptococcus uberis clinical mastitis at cow level could be predicted from the historical presence of specific strains of S. uberis on dairy farms. Matrix-assisted laser desorption ionization time of flight mass spectrometry was used to identify S. uberis isolates potentially capable of contagious transmission. Data were available from 10,652 cows from 52 English and Welsh dairy farms over a 14 month period, and 521 isolates of S. uberis from clinical mastitis cases were available for analysis. As well as the temporal herd history of clinical mastitis associated with particular S. uberis strains, other exposure variables included cow parity, stage of lactation, milk yield, and somatic cell count. Observations were structured longitudinally as repeated weekly measures through the study period for each cow. Data were analyzed in a Bayesian framework using multilevel logistic regression models. Similarity of mass spectral profiles between isolates of S. uberis from consecutive clinical cases of mastitis in herds was used to indicate potential for contagious phenotypic characteristics. Cross validation showed that new isolates with these characteristics could be identified with an accuracy of 90% based on bacterial protein mass spectral characteristics alone. The cow-level risk in any week of these S. uberis clinical mastitis cases increased with the presence of the same specific strains of S. uberis in other cows in the herd during the previous 2 weeks. The final statistical model indicated there would be a 2-3 fold increase in the risk of S. uberis clinical mastitis associated with particular strains if these occurred in the herd 1 and 2 weeks previously. The results suggest that specific strains of S. uberis may be involved with contagious transmission, and predictions based on their occurrence could be used as an early warning surveillance system to enhance the control of S. uberis mastitis. Copyright

  12. Identification of Coagulase-Negative Staphylococci from Bovine Intramammary Infection by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Gonçalves, Juliano Leonel; Barreiro, Juliana Regina; Braga, Patrícia Aparecida de Campos; Prada e Silva, Luis Felipe; Eberlin, Marcos Nogueira

    2014-01-01

    Coagulase-negative staphylococci (CoNS) are among the main pathogens causing bovine intramammary infection (IMI) in many countries. However, one of the limitations related to the specific diagnosis of CoNS is the lack of an accurate, rapid, and convenient method that can differentiate the bacterial species comprising this group. The aim of this study was to evaluate the ability of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) to accurately identify CoNS species in dairy cow IMI. In addition, the study aimed to determine the frequency of CoNS species causing bovine IMI. A total of 108 bacterial isolates were diagnosed as CoNS by microbiological cultures from two milk samples collected from 21 dairy herds; the first sample was collected at the cow level (i.e., 1,242 composite samples from all quarters), while the second sample was collected at the mammary quarter level (i.e., 1,140 mammary samples collected from 285 cows). After CoNS isolation was confirmed by microbiological culture for both samples, all CoNS isolates (n = 108) were genotypically differentiated by PCR restriction fragment length polymorphism (RFLP) analysis of a partial groEL gene sequence and subjected to the MALDI-TOF MS identification procedure. MALDI-TOF MS correctly identified 103 (95.4%) of the CoNS isolates identified by PCR-RFLP at the species level. Eleven CoNS species isolated from bovine IMI were identified by PCR-RFLP, and the most prevalent species was Staphylococcus chromogenes (n = 80; 74.1%). In conclusion, MALDI-TOF MS may be a reliable alternative method for differentiating CoNS species causing bovine IMI. PMID:24622096

  13. Protein secondary structure and stability determined by combining exoproteolysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Villanueva, Josep; Villegas, Virtudes; Querol, Enrique; Avilés, Francesc X; Serrano, Luis

    2002-09-01

    In the post-genomic era, several projects focused on the massive experimental resolution of the three-dimensional structures of all the proteins of different organisms have been initiated. Simultaneously, significant progress has been made in the ab initio prediction of protein three-dimensional structure. One of the keys to the success of such a prediction is the use of local information (i.e. secondary structure). Here we describe a new limited proteolysis methodology, based on the use of unspecific exoproteases coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), to map quickly secondary structure elements of a protein from both ends, the N- and C-termini. We show that the proteolytic patterns (mass spectra series) obtained can be interpreted in the light of the conformation and local stability of the analyzed proteins, a direct correlation being observed between the predicted and the experimentally derived protein secondary structure. Further, this methodology can be easily applied to check rapidly the folding state of a protein and characterize mutational effects on protein conformation and stability. Moreover, given global stability information, this methodology allows one to locate the protein regions of increased or decreased conformational stability. All of this can be done with a small fraction of the amount of protein required by most of the other methods for conformational analysis. Thus limited exoproteolysis, together with MALDI-TOF MS, can be a useful tool to achieve quickly the elucidation of protein structure and stability. Copyright 2002 John Wiley & Sons, Ltd.

  14. Identification of proteins in a human pleural exudate using two-dimensional preparative liquid-phase electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry.

    Nilsson, C L; Puchades, M; Westman, A; Blennow, K; Davidsson, P

    1999-01-01

    Pleural effusion may occur in patients suffering from physical trauma or systemic disorders such as infection, inflammation, or cancer. In order to investigate proteins in a pleural exudate from a patient with severe pneumonia, we used a strategy that combined preparative two-dimensional liquid-phase electrophoresis (2-D LPE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and Western blotting. Preparative 2-D LPE is based on the same principles as analytical 2-D gel electrophoresis, except that the proteins remain in liquid phase during the entire procedure. In the first dimension, liquid-phase isoelectric focusing allows for the enrichment of proteins in liquid fractions. In the Rotofor cell, large volumes (up to 55 mL) and protein amounts (up to 1-2 g) can be loaded. Several low abundance proteins, cystatin C, haptoglobin, transthyretin, beta2-microglobulin, and transferrin, were detected after liquid-phase isoelectric focusing, through Western blotting analysis, in a pleural exudate (by definition, >25 g/L total protein). Direct MALDI-TOF-MS analysis of proteins in a Rotofor fraction is demonstrated as well. MALDI-TOF-MS analysis of a tryptic digest of a continuous elution sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) fraction confirmed the presence of cystatin C. By applying 2-D LPE, MALDI-TOF-MS, and Western blotting to the analysis of this pleural exudate, we were able to confirm the identity of proteins of potential diagnostic value. Our findings serve to illustrate the usefulness of this combination of methods in the analysis of pathological fluids.

  15. Rapid, simple, and highly sensitive analysis of drugs in biological samples using thin-layer chromatography coupled with matrix-assisted laser desorption/ionization mass spectrometry.

    Kuwayama, Kenji; Tsujikawa, Kenji; Miyaguchi, Hajime; Kanamori, Tatsuyuki; Iwata, Yuko T; Inoue, Hiroyuki

    2012-01-01

    Rapid and precise identification of toxic substances is necessary for urgent diagnosis and treatment of poisoning cases and for establishing the cause of death in postmortem examinations. However, identification of compounds in biological samples using gas chromatography and liquid chromatography coupled with mass spectrometry entails time-consuming and labor-intensive sample preparations. In this study, we examined a simple preparation and highly sensitive analysis of drugs in biological samples such as urine, plasma, and organs using thin-layer chromatography coupled with matrix-assisted laser desorption/ionization mass spectrometry (TLC/MALDI/MS). When the urine containing 3,4-methylenedioxymethamphetamine (MDMA) without sample dilution was spotted on a thin-layer chromatography (TLC) plate and was analyzed by TLC/MALDI/MS, the detection limit of the MDMA spot was 0.05 ng/spot. The value was the same as that in aqueous solution spotted on a stainless steel plate. All the 11 psychotropic compounds tested (MDMA, 4-hydroxy-3-methoxymethamphetamine, 3,4-methylenedioxyamphetamine, methamphetamine, p-hydroxymethamphetamine, amphetamine, ketamine, caffeine, chlorpromazine, triazolam, and morphine) on a TLC plate were detected at levels of 0.05-5 ng, and the type (layer thickness and fluorescence) of TLC plate did not affect detection sensitivity. In addition, when rat liver homogenate obtained after MDMA administration (10 mg/kg) was spotted on a TLC plate, MDMA and its main metabolites were identified using TLC/MALDI/MS, and the spots on a TLC plate were visualized by MALDI/imaging MS. The total analytical time from spotting of intact biological samples to the output of analytical results was within 30 min. TLC/MALDI/MS enabled rapid, simple, and highly sensitive analysis of drugs from intact biological samples and crude extracts. Accordingly, this method could be applied to rapid drug screening and precise identification of toxic substances in poisoning cases and

  16. Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of Molds of the Fusarium Genus

    Stubbe, Dirk; De Cremer, Koen; Piérard, Denis; Normand, Anne-Cécile; Piarroux, Renaud; Detandt, Monique; Hendrickx, Marijke

    2014-01-01

    The rates of infection with Fusarium molds are increasing, and a diverse number of Fusarium spp. belonging to different species complexes can cause infection. Conventional species identification in the clinical laboratory is time-consuming and prone to errors. We therefore evaluated whether matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a useful alternative. The 289 Fusarium strains from the Belgian Coordinated Collections of Microorganisms (BCCM)/Institute of Hygiene and Epidemiology Mycology (IHEM) culture collection with validated sequence-based identities and comprising 40 species were used in this study. An identification strategy was developed, applying a standardized MALDI-TOF MS assay and an in-house reference spectrum database. In vitro antifungal testing was performed to assess important differences in susceptibility between clinically relevant species/species complexes. We observed that no incorrect species complex identifications were made by MALDI-TOF MS, and 82.8% of the identifications were correct to the species level. This success rate was increased to 91% by lowering the cutoff for identification. Although the identification of the correct species complex member was not always guaranteed, antifungal susceptibility testing showed that discriminating between Fusarium species complexes can be important for treatment but is not necessarily required between members of a species complex. With this perspective, some Fusarium species complexes with closely related members can be considered as a whole, increasing the success rate of correct identifications to 97%. The application of our user-friendly MALDI-TOF MS identification approach resulted in a dramatic improvement in both time and accuracy compared to identification with the conventional method. A proof of principle of our MALDI-TOF MS approach in the clinical setting using recently isolated Fusarium strains demonstrated its validity. PMID:25411180

  17. Matrix-assisted laser desorption/ionization mass spectrometry imaging of cell cultures for the lipidomic analysis of potential lipid markers in human breast cancer invasion.

    Wang, Shujuan; Chen, Xiaowu; Luan, Hemi; Gao, Dan; Lin, Shuhai; Cai, Zongwei; Liu, Jianjun; Liu, Hongxia; Jiang, Yuyang

    2016-02-28

    Breast cancer is the leading cause of cancer death among women worldwide. Identification of lipid targets that play a role in breast cancer invasion may advance our understanding of the rapid progression of cancer and may lead to the development of new biomarkers for the disease. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) was applied for the lipidomic profiling of two poorly invasive and two highly invasive breast cancer cell lines to identify the differentially accumulated lipids related to the invasive phenotype. The four cell lines were individually grown on indium tin oxide (ITO)-coated glass slides, analyzed as cell cultures. The raster width and matrix for detection were optimized to improve detection sensitivity. Optimized MSI measurements were performed directly on the cell culture with 9-aminoacridine as matrix, resulting in 215 endogenous compounds detected in positive ion mode and 267 endogenous compounds in negative ion mode in all the four cell lines, representing the largest group of analytes that have been analyzed from cells by a single MSI study. In highly invasive cell lines, 31 lipids including phosphatidylglycerol (PG) and phosphatidic acids were found upregulated and eight lipids including sphingomyelin (SM) downregulated in negative ion mode. The products of de novo fatty acid synthesis incorporated into membrane phospholipids, like oleic-acid-containing PG, may be involved in mitochondrial dysfunction and thus affect the invasion of breast cancer cells. The deficiency of SM may be related to the disruption of apoptosis in highly invasive cancer cells. This work uncovered more analytes in cells by MSI than previous reports, providing a better visualization and novel insights to advance our understanding of the relationship between rapid progression of breast cancer and lipid metabolism. The most altered lipids may aid the discovery of diagnostic markers and therapeutic targets of breast cancer. Copyright

  18. Identification of Enterobacteriaceae by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using the VITEK MS system.

    Richter, S S; Sercia, L; Branda, J A; Burnham, C-A D; Bythrow, M; Ferraro, M J; Garner, O B; Ginocchio, C C; Jennemann, R; Lewinski, M A; Manji, R; Mochon, A B; Rychert, J A; Westblade, L F; Procop, G W

    2013-12-01

    This multicenter study evaluated the accuracy of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry identifications from the VITEK MS system (bioMérieux, Marcy l'Etoile, France) for Enterobacteriaceae typically encountered in the clinical laboratory. Enterobacteriaceae isolates (n = 965) representing 17 genera and 40 species were analyzed on the VITEK MS system (database v2.0), in accordance with the manufacturer's instructions. Colony growth (≤72 h) was applied directly to the target slide. Matrix solution (α-cyano-4-hydroxycinnamic acid) was added and allowed to dry before mass spectrometry analysis. On the basis of the confidence level, the VITEK MS system provided a species, genus only, or no identification for each isolate. The accuracy of the mass spectrometric identification was compared to 16S rRNA gene sequencing performed at MIDI Labs (Newark, DE). Supplemental phenotypic testing was performed at bioMérieux when necessary. The VITEK MS result agreed with the reference method identification for 96.7% of the 965 isolates tested, with 83.8% correct to the species level and 12.8% limited to a genus-level identification. There was no identification for 1.7% of the isolates. The VITEK MS system misidentified 7 isolates (0.7 %) as different genera. Three Pantoea agglomerans isolates were misidentified as Enterobacter spp. and single isolates of Enterobacter cancerogenus, Escherichia hermannii, Hafnia alvei, and Raoultella ornithinolytica were misidentified as Klebsiella oxytoca, Citrobacter koseri, Obesumbacterium proteus, and Enterobacter aerogenes, respectively. Eight isolates (0.8 %) were misidentified as a different species in the correct genus. The VITEK MS system provides reliable mass spectrometric identifications for Enterobacteriaceae.

  19. Multicenter Evaluation of the Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System for Identification of Gram-Positive Aerobic Bacteria

    Burnham, Carey-Ann D.; Bythrow, Maureen; Garner, Omai B.; Ginocchio, Christine C.; Jennemann, Rebecca; Lewinski, Michael A.; Manji, Ryhana; Mochon, A. Brian; Procop, Gary W.; Richter, Sandra S.; Sercia, Linda; Westblade, Lars F.; Ferraro, Mary Jane; Branda, John A.

    2013-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF) is gaining momentum as a tool for bacterial identification in the clinical microbiology laboratory. Compared with conventional methods, this technology can more readily and conveniently identify a wide range of organisms. Here, we report the findings from a multicenter study to evaluate the Vitek MS v2.0 system (bioMérieux, Inc.) for the identification of aerobic Gram-positive bacteria. A total of 1,146 unique isolates, representing 13 genera and 42 species, were analyzed, and results were compared to those obtained by nucleic acid sequence-based identification as the reference method. For 1,063 of 1,146 isolates (92.8%), the Vitek MS provided a single identification that was accurate to the species level. For an additional 31 isolates (2.7%), multiple possible identifications were provided, all correct at the genus level. Mixed-genus or single-choice incorrect identifications were provided for 18 isolates (1.6%). Although no identification was obtained for 33 isolates (2.9%), there was no specific bacterial species for which the Vitek MS consistently failed to provide identification. In a subset of 463 isolates representing commonly encountered important pathogens, 95% were accurately identified to the species level and there were no misidentifications. Also, in all but one instance, the Vitek MS correctly differentiated Streptococcus pneumoniae from other viridans group streptococci. The findings demonstrate that the Vitek MS system is highly accurate for the identification of Gram-positive aerobic bacteria in the clinical laboratory setting. PMID:23658261

  20. Multicenter evaluation of the Vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system for identification of Gram-positive aerobic bacteria.

    Rychert, Jenna; Burnham, Carey-Ann D; Bythrow, Maureen; Garner, Omai B; Ginocchio, Christine C; Jennemann, Rebecca; Lewinski, Michael A; Manji, Ryhana; Mochon, A Brian; Procop, Gary W; Richter, Sandra S; Sercia, Linda; Westblade, Lars F; Ferraro, Mary Jane; Branda, John A

    2013-07-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) is gaining momentum as a tool for bacterial identification in the clinical microbiology laboratory. Compared with conventional methods, this technology can more readily and conveniently identify a wide range of organisms. Here, we report the findings from a multicenter study to evaluate the Vitek MS v2.0 system (bioMérieux, Inc.) for the identification of aerobic Gram-positive bacteria. A total of 1,146 unique isolates, representing 13 genera and 42 species, were analyzed, and results were compared to those obtained by nucleic acid sequence-based identification as the reference method. For 1,063 of 1,146 isolates (92.8%), the Vitek MS provided a single identification that was accurate to the species level. For an additional 31 isolates (2.7%), multiple possible identifications were provided, all correct at the genus level. Mixed-genus or single-choice incorrect identifications were provided for 18 isolates (1.6%). Although no identification was obtained for 33 isolates (2.9%), there was no specific bacterial species for which the Vitek MS consistently failed to provide identification. In a subset of 463 isolates representing commonly encountered important pathogens, 95% were accurately identified to the species level and there were no misidentifications. Also, in all but one instance, the Vitek MS correctly differentiated Streptococcus pneumoniae from other viridans group streptococci. The findings demonstrate that the Vitek MS system is highly accurate for the identification of Gram-positive aerobic bacteria in the clinical laboratory setting.

  1. Specific Cα-C Bond Cleavage of β-Carbon-Centered Radical Peptides Produced by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

    Nagoshi, Keishiro; Yamakoshi, Mariko; Sakamoto, Kenya; Takayama, Mitsuo

    2018-04-01

    Radical-driven dissociation (RDD) of hydrogen-deficient peptide ions [M - H + H]·+ has been examined using matrix-assisted laser dissociation/ionization in-source decay mass spectrometry (MALDI-ISD MS) with the hydrogen-abstracting matrices 4-nitro-1-naphthol (4,1-NNL) and 5-nitrosalicylic acid (5-NSA). The preferential fragment ions observed in the ISD spectra include N-terminal [a] + ions and C-terminal [x]+, [y + 2]+, and [w]+ ions which imply that β-carbon (Cβ)-centered radical peptide ions [M - Hβ + H]·+ are predominantly produced in MALDI conditions. RDD reactions from the peptide ions [M - Hβ + H]·+ successfully explains the fact that both [a]+ and [x]+ ions arising from cleavage at the Cα-C bond of the backbone of Gly-Xxx residues are missing from the ISD spectra. Furthermore, the formation of [a]+ ions originating from the cleavage of Cα-C bond of deuterated Ala(d3)-Xxx residues indicates that the [a]+ ions are produced from the peptide ions [M - Hβ + H]·+ generated by deuteron-abstraction from Ala(d3) residues. It is suggested that from the standpoint of hydrogen abstraction via direct interactions between the nitro group of matrix and hydrogen of peptides, the generation of the peptide radical ions [M - Hβ + H]·+ is more favorable than that of the α-carbon (Cα)-centered radical ions [M - Hα + H]·+ and the amide nitrogen-centered radical ions [M - HN + H]·+, while ab initio calculations indicate that the formation of [M - Hα + H]·+ is energetically most favorable. [Figure not available: see fulltext.

  2. Matrix-assisted laser-desorption/ionization mass spectrometric imaging of olanzapine in a single hair using esculetin as a matrix.

    Wang, Hang; Wang, Ying; Wang, Ge; Hong, Lizhi

    2017-07-15

    Matrix-assisted laser desorption/ionization-mass spectrometric imaging (MALDI-MSI) for the analysis of intact hair is a powerful tool for monitoring changes in drug consumption. The embedding of a low drug concentration in the hydrophobic hair matrix makes it difficult to extract and detect, and requires an improved method to increase detection sensitivity. In this study, an MSI method using MALDI-Fourier transform ion cyclotron resonance was developed for direct identification and imaging of olanzapine in hair samples using the positive ion mode. Following decontamination, scalp hair samples from an olanzapine user were scraped from the proximal to the distal end three times, and 5mm hair sections were fixed onto an Indium-Tin-Oxide (ITO)-coated microscopic glass slide. Esculetin (6,7-dihydroxy-2H-chromen-2-one) was used as a new hydrophobic matrix to increase the affinity, extraction and ionization efficiency of olanzapine in the hair samples. The spatial distribution of olanzapine was observed using five single hairs from the same drug user. This matrix improves the affinity of olanzapine in hair for molecular imaging with mass spectrometry. This method may provide a detection power for olanzapine to the nanogram level per 5mm hair. Time course changes in the MSI results were also compared with quantitative HPLC-MS/MS for each 5mm segment of single hair shafts selected from the MALDI target. MALDI imaging intensities in single hairs showed good semi-quantitative correlation with the results from conventional HPLC-MS/MS. MALDI-MSI is suitable for monitoring drug intake with a high time resolution. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Comparison of Vitek Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Versus Conventional Methods in Candida Identification.

    Keçeli, Sema Aşkın; Dündar, Devrim; Tamer, Gülden Sönmez

    2016-02-01

    Candida species are generally identified by conventional methods such as germ tube or morphological appearance on corn meal agar, biochemical methods using API kits and molecular biological methods. Alternative to these methods, rapid and accurate identification methods of microorganisms called matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDİ-TOF MS) has recently been described. In this study, Candida identification results by API Candida kit, API 20C AUX kit and identifications on corn meal agar (CMA) are compared with the results obtained on Vitek-MS. All results were confirmed by sequencing internal transcribed spacer (ITS) regions of rDNA. Totally, 97 Candida strains were identified by germ tube test, CMA, API and Vitek-MS. Vitek-MS results were compatible with 74.2 % of API 20C AUX and 81.4 % of CMA results. The difference between the results of API Candida and API 20C AUX was detected. The ratio of discrepancy between Vitek-MS and API 20C AUX was 25.8 %. Candida species mostly identified as C. famata or C. tropicalis by and not compatible with API kits were identified as C. albicans by Vitek-MS. Sixteen Candida species having discrepant results with Vitek-MS, API or CMA were randomly chosen, and ITS sequence analysis was performed. The results of sequencing were compatible 56.2 % with API 20C AUX, 50 % with CMA and 93.7 % with Vitek-MS. When compared with conventional identification methods, MS results are more reliable and rapid for Candida identification. MS system may be used as routine identification method in clinical microbiology laboratories.

  4. Examination of the skin barrier repair/wound healing process using a living skin equivalent model and matrix-assisted laser desorption-ionization-mass spectrometry imaging.

    Lewis, E E L; Barrett, M R T; Freeman-Parry, L; Bojar, R A; Clench, M R

    2018-04-01

    Examination of the skin barrier repair/wound healing process using a living skin equivalent (LSE) model and matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) to identify lipids directly involved as potential biomarkers. These biomarkers may be used to determine whether an in vivo wound is going to heal for example if infected. An in vitro LSE model was wounded with a scalpel blade and assessed at day 4 post-wounding by histology and MALDI-MSI. Samples were sectioned at wound site and were either formalin-fixed paraffin-embedded (FFPE) for histology or snapped frozen (FF) for MSI analysis. The combination of using an in vitro wounded skin model with MSI allowed the identification of lipids involved in the skin barrier repair/wound healing process. The technique was able to highlight lipids directly in the wound site and distinguish differences in lipid distribution between the epidermis and wound site. This novel method of coupling an in vitro LSE with MSI allowed in-depth molecular analysis of the skin barrier repair/wound healing process. The technique allowed the identification of lipids directly involved in the skin barrier repair/wound healing process, indicating these biomarkers may be potentially be used within the clinic. These biomarkers will help to determine, which stage of the skin barrier repair/wound healing process the wound is in to provide the best treatment. © 2018 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  5. Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    De Carolis, E; Posteraro, B; Lass-Flörl, C; Vella, A; Florio, A R; Torelli, R; Girmenia, C; Colozza, C; Tortorano, A M; Sanguinetti, M; Fadda, G

    2012-05-01

    Accurate species discrimination of filamentous fungi is essential, because some species have specific antifungal susceptibility patterns, and misidentification may result in inappropriate therapy. We evaluated matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification through direct surface analysis of the fungal culture. By use of culture collection strains representing 55 species of Aspergillus, Fusarium and Mucorales, a reference database was established for MALDI-TOF MS-based species identification according to the manufacturer's recommendations for microflex measurements and MALDI BioTyper 2.0 software. The profiles of young and mature colonies were analysed for each of the reference strains, and species-specific spectral fingerprints were obtained. To evaluate the database, 103 blind-coded fungal isolates collected in the routine clinical microbiology laboratory were tested. As a reference method for species designation, multilocus sequencing was used. Eighty-five isolates were unequivocally identified to the species level (≥99% sequence similarity); 18 isolates producing ambiguous results at this threshold were initially rated as identified to the genus level only. Further molecular analysis definitively assigned these isolates to the species Aspergillus oryzae (17 isolates) and Aspergillus flavus (one isolate), concordant with the MALDI-TOF MS results. Excluding nine isolates that belong to the fungal species not included in our reference database, 91 (96.8%) of 94 isolates were identified by MALDI-TOF MS to the species level, in agreement with the results of the reference method; three isolates were identified to the genus level. In conclusion, MALDI-TOF MS is suitable for the routine identification of filamentous fungi in a medical microbiology laboratory. © 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.

  6. Reliable and reproducible method for rapid identification of Nocardia species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Toyokawa, Masahiro; Kimura, Keigo; Nishi, Isao; Sunada, Atsuko; Ueda, Akiko; Sakata, Tomomi; Asari, Seishi

    2013-01-01

    Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been challenged for the identification of Nocardia species. However, the standard ethanol-formic acid extraction alone is insufficient in allowing the membrane proteins of Nocardia species to be ionized by the matrix. We therefore aimed to establish our new extraction method for the MALDI-TOF MS-based identification of Nocardia species isolates. Our modified extraction procedure is through dissociation in 0.5% Tween-20 followed by bacterial heat-inactivation, mechanical breaking of the cell wall by acid-washed glass beads and protein extraction with formic acid and acetonitrile. As reference methods for species identification, full-length 16S rRNA gene sequencing and some phenotypical tests were used. In a first step, we made our own Nocardia database by analyzing 13 strains (13 different species including N. elegans, N. otitidiscaviarum, N. asiatica, N. abscessus, N. brasiliensis, N. thailandica, N. farcinica, N. nova, N. mikamii, N. cyriacigeorgica, N. asteroids, Nocardiopsis alba, and Micromonospora sp.) and registered to the MALDI BioTyper database. Then we established our database. The analysis of 12 challenge strains using the our database gave a 100% correct identification, including 8 strains identified to the species level and 4 strains to the genus level (N. elegans, N. nova, N. farcinica, Micromonospora sp.) according to the manufacture's log score specifications. In the estimation of reproducibility of our method intended for 4 strains, both within-run and between-run reproducibility were excellent. These data indicates that our method for rapid identification of Nocardia species is with reliability, reproducibility and cost effective.

  7. Matrix-assisted laser desorption/ionization mass spectrometric imaging for the rapid segmental analysis of methamphetamine in a single hair using umbelliferone as a matrix.

    Wang, Hang; Wang, Ying

    2017-07-04

    Segmental hair analysis offers a longer period for retrospective drug detection than blood or urine. Hair is a keratinous fiber and is strongly hydrophobic. The embedding of drugs in hydrophobic hair at low concentrations makes it difficult for extraction and detection with matrix-assisted laser desorption/ionization (MALDI) coupled with mass spectrometric imaging (MSI). In this study, a single scalp hair was longitudinally cut with a cryostat section to a length of 4 mm and fixed onto a stainless steel MALDI plate. Umbelliferone was used as a new hydrophobic matrix to enrich and assist the ionization efficiency of methamphetamine in the hair sample. MALDI-Fourier transform ion cyclotron resonance (FTICR)-MS profiling and imaging were performed for direct detection and mapping of methamphetamine on the longitudinal sections of the single hair sample in positive ion mode. Using MALDI-MSI, the distribution of methamphetamine was observed throughout five longitudinally sectioned hair samples from a drug abuser. The changes of methamphetamine were also semi-quantified by comparing the ratios of methamphetamine/internal standard (I.S). This method improves the detection sensitivity of target drugs embedded in a hair matrix for imaging with mass spectrometry. The method could provide a detection level of methamphetamine down to a nanogram per milligram incorporated into hair. The results were also compared with the conventional high performance liquid chromatography -tandem mass spectrometry (HPLC-MS/MS) method. Changes in the imaging results over time by the MSI method showed good semi-quantitative correlation to the results from the HPLC-MS/MS method. This study provides a powerful tool for drug abuse control and forensic medicine analysis in a narrow time frame, and a reduction in the sample amount required. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Cost Savings Realized by Implementation of Routine Microbiological Identification by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Tran, Anthony; Alby, Kevin; Kerr, Alan; Jones, Melissa; Gilligan, Peter H

    2015-08-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is an emerging technology for rapid identification of bacterial and fungal isolates. In comparison to conventional methods, this technology is much less labor intensive and can provide accurate and reliable results in minutes from a single isolated colony. We compared the cost of performing the bioMérieux Vitek MALDI-TOF MS with conventional microbiological methods to determine the amount saved by the laboratory by converting to the new technology. Identification costs for 21,930 isolates collected between April 1, 2013, and March 31, 2014, were directly compared for MALDI-TOF MS and conventional methodologies. These isolates were composed of commonly isolated organisms, including commonly encountered aerobic and facultative bacteria and yeast but excluding anaerobes and filamentous fungi. Mycobacterium tuberculosis complex and rapidly growing mycobacteria were also evaluated for a 5-month period during the study. Reagent costs and a total cost analysis that included technologist time in addition to reagent expenses and maintenance service agreement costs were analyzed as part of this study. The use of MALDI-TOF MS equated to a net savings of $69,108.61, or 87.8%, in reagent costs annually compared to traditional methods. When total costs are calculated to include technologist time and maintenance costs, traditional identification would have cost $142,532.69, versus $68,886.51 with the MALDI-TOF MS method, resulting in a laboratory savings of $73,646.18, or 51.7%, annually by adopting the new technology. The initial cost of the instrument at our usage level would be offset in about 3 years. MALDI-TOF MS not only represents an innovative technology for the rapid and accurate identification of bacterial and fungal isolates, it also provides a significant cost savings for the laboratory. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Cost Analysis of Implementing Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Plus Real-Time Antimicrobial Stewardship Intervention for Bloodstream Infections.

    Patel, Twisha S; Kaakeh, Rola; Nagel, Jerod L; Newton, Duane W; Stevenson, James G

    2017-01-01

    Studies evaluating rapid diagnostic testing plus stewardship intervention have consistently demonstrated improved clinical outcomes for patients with bloodstream infections. However, the cost of implementing new rapid diagnostic testing can be significant, and such testing usually does not generate additional revenue. There are minimal data evaluating the impact of adding matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for rapid organism identification and dedicating pharmacy stewardship personnel time on the total hospital costs. A cost analysis was performed utilizing patient data generated from the hospital cost accounting system and included additional costs of MALDI-TOF equipment, supplies and personnel, and dedicated pharmacist time for blood culture review and of making interventions to antimicrobial therapy. The cost analysis was performed from a hospital perspective for 3-month blocks before and after implementation of MALDI-TOF plus stewardship intervention. A total of 480 patients with bloodstream infections were included in the analysis: 247 in the preintervention group and 233 in the intervention group. Thirty-day mortality was significantly improved in the intervention group (12% versus 21%, P cost per bloodstream infection was lower in the intervention group ($42,580 versus $45,019). Intensive care unit cost per bloodstream infection accounted for the largest share of the total costs in each group and was also lower in the intervention group ($10,833 versus $13,727). Implementing MALDI-TOF plus stewardship review and intervention decreased mortality for patients with bloodstream infections. Despite the additional costs of implementing MALDI-TOF and of dedicating pharmacy stewardship personnel time to interventions, the total hospital costs decreased by $2,439 per bloodstream infection, for an approximate annual cost savings of $2.34 million. Copyright © 2016 American Society for Microbiology.

  10. Identification of bacteria in blood culture broths using matrix-assisted laser desorption-ionization Sepsityper™ and time of flight mass spectrometry.

    Kok, Jen; Thomas, Lee C; Olma, Thomas; Chen, Sharon C A; Iredell, Jonathan R

    2011-01-01

    Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) is a novel method for the direct identification of bacteria from blood culture broths. We evaluate for the first time, the performance of the MALDI Sepsityper™ Kit and MS for the identification of bacteria compared to standard phenotypic methods using the manufacturer's specified bacterial identification criteria (spectral scores ≥1.700-1.999 and ≥2.000 indicated identification to genus and species level, respectively). Five hundred and seven positive blood culture broths were prospectively examined, of which 379 (74.8%; 358 monomicrobial, 21 polymicrobial) were identified by MALDI-TOF MS; 195 (100%) and 132 (67.7%) of 195 gram-positive; and 163 (100%) and 149 (91.4%) of 163 gram-negative organisms from monomicrobial blood cultures were correctly identified to genus and species level, respectively. Spectral scores blood culture broths, including 31.6% and 32.3% of gram-positive and polymicrobial blood cultures, respectively. Significantly more gram-negative organisms were identified compared to gram-positive organisms at species level (pblood cultures were misidentified, but at species level only; including four monomicrobial blood cultures with Streptococcus oralis/mitis that were misidentified as Streptococcus pneumoniae. Positive predictive values for the direct identification of both gram-positive and gram-negative bacteria from monomicrobial blood culture broths to genus level were 100%. A diagnostic algorithm for positive blood culture broths that incorporates gram staining and MALDI-TOF MS should identify the majority of pathogens, particularly to genus level.

  11. Direct identification of bacteria from charcoal-containing blood culture bottles using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry.

    Wüppenhorst, N; Consoir, C; Lörch, D; Schneider, C

    2012-10-01

    Several protocols for direct matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) from positive blood cultures are currently used to speed up the diagnostic process of bacteraemia. Identification rates are high and results are accurate for the BACTEC™ system and for charcoal-free bottles. Only a few studies have evaluated protocols for charcoal-containing BacT/ALERT bottles reaching substantially lower identification rates. We established a new protocol for sample preparation from aerobic and anaerobic positive charcoal-containing BacT/ALERT blood culture bottles and measured the protein profiles (n = 167). Then, we integrated this protocol in the routine workflow of our laboratory (n = 212). During the establishment of our protocol, 74.3 % of bacteria were correctly identified to the species level, in 23.4 %, no result and in 2.4 %, a false identification were obtained. Reliable criteria for correct species identification were a score value ≥1.400 and a best match on rank 1-3 of the same species. Identification rates during routine workflow were 77.8 % for correct identification, 20.8 % for not identified samples and 1.4 % for discordant identification. In conclusion, our results indicate that MALDI-TOF MS is possible, even from charcoal-containing blood cultures. Reliable criteria for correct species identification are a score value ≥1.400 and a best match on rank 1-3 of a single species.

  12. Direct bacterial identification in positive blood cultures by use of two commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry systems.

    Chen, Jonathan H K; Ho, Pak-Leung; Kwan, Grace S W; She, Kevin K K; Siu, Gilman K H; Cheng, Vincent C C; Yuen, Kwok-Yung; Yam, Wing-Cheong

    2013-06-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of bacteria and fungi was recently introduced in microbiology laboratories. This technology could greatly improve the clinical management of patients and guidance for chemotherapy. In this study, we used a commercial MALDI Sepsityper extraction method to evaluate the performance of two commercial MALDI-TOF MS systems, the Vitek MS IVD (bioMérieux) and the Microflex LT Biotyper (Bruker Daltonics) for direct bacterial identification in positive blood cultures. In 181 monomicrobial cultures, both systems generated genus to species level identifications for >90% of the specimens (Biotyper, 177/181 [97.8%]; Vitek MS IVD, 167/181 [92.3%]). Overall, the Biotyper system generated significantly more accurate identifications than the Vitek MS IVD system (P = 0.016; 177 versus 167 out of 181 specimens). The Biotyper system identified the minority species among polymicrobial blood cultures. We also compared the performance of an in-house extraction method with that of the Sepsityper on both MALDI-TOF MS systems. The in-house method generated more correct identifications at the genus level than the Sepsityper (96.7% versus 93.5%) on the Biotyper system, whereas the two methods exhibited the same performance level (88.0% versus 88.0%) on the Vitek MS IVD system. Our study confirmed the practical advantages of MALDI-TOF MS, and our in-house extraction method reduced the reagent cost to $1 per specimen, with a shorter turnaround time of 3 h, which is highly cost-effective for a diagnostic microbiology service.

  13. Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry.

    Bernard La Scola

    Full Text Available BACKGROUND: With long delays observed between sampling and availability of results, the usefulness of blood cultures in the context of emergency infectious diseases has recently been questioned. Among methods that allow quicker bacterial identification from growing colonies, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF mass spectrometry was demonstrated to accurately identify bacteria routinely isolated in a clinical biology laboratory. In order to speed up the identification process, in the present work we attempted bacterial identification directly from blood culture bottles detected positive by the automate. METHODOLOGY/PRINCIPAL FINDINGS: We prospectively analysed routine MALDI-TOF identification of bacteria detected in blood culture by two different protocols involving successive centrifugations and then lysis by trifluoroacetic acid or formic acid. Of the 562 blood culture broths detected as positive by the automate and containing one bacterial species, 370 (66% were correctly identified. Changing the protocol from trifluoroacetic acid to formic acid improved identification of Staphylococci, and overall correct identification increased from 59% to 76%. Lack of identification was observed mostly with viridans streptococci, and only one false positive was observed. In the 22 positive blood culture broths that contained two or more different species, only one of the species was identified in 18 samples, no species were identified in two samples and false species identifications were obtained in two cases. The positive predictive value of bacterial identification using this procedure was 99.2%. CONCLUSIONS/SIGNIFICANCE: MALDI-TOF MS is an efficient method for direct routine identification of bacterial isolates in blood culture, with the exception of polymicrobial samples and viridans streptococci. It may replace routine identification performed on colonies, provided improvement for the specificity of blood culture

  14. Direct identification of pathogens from positive blood cultures using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry.

    Rodríguez-Sánchez, B; Sánchez-Carrillo, C; Ruiz, A; Marín, M; Cercenado, E; Rodríguez-Créixems, M; Bouza, E

    2014-07-01

    In recent years, matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) has proved a rapid and reliable method for the identification of bacteria and yeasts that have already been isolated. The objective of this study was to evaluate this technology as a routine method for the identification of microorganisms directly from blood culture bottles (BCBs), before isolation, in a large collection of samples. For this purpose, 1000 positive BCBs containing 1085 microorganisms have been analysed by conventional phenotypic methods and by MALDI-TOF MS. Discrepancies have been resolved using molecular methods: the amplification and sequencing of the 16S rRNA gene or the Superoxide Dismutase gene (sodA) for streptococcal isolates. MALDI-TOF predicted a species- or genus-level identification of 81.4% of the analysed microorganisms. The analysis by episode yielded a complete identification of 814 out of 1000 analysed episodes (81.4%). MALDI-TOF identification is available for clinicians within hours of a working shift, as oppose to 18 h later when conventional identification methods are performed. Moreover, although further improvement of sample preparation for polymicrobial BCBs is required, the identification of more than one pathogen in the same BCB provides a valuable indication of unexpected pathogens when their presence may remain undetected in Gram staining. Implementation of MALDI-TOF identification directly from the BCB provides a rapid and reliable identification of the causal pathogen within hours. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

  15. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry identification of large colony beta-hemolytic streptococci containing Lancefield groups A, C, and G.

    Jensen, Christian Salgård; Dam-Nielsen, Casper; Arpi, Magnus

    2015-08-01

    The aim of this study was to investigate whether large colony beta-hemolytic streptococci containing Lancefield groups A, C, and G can be adequately identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-ToF). Previous studies show varying results, with an identification rate from below 50% to 100%. Large colony beta-hemolytic streptococci containing Lancefield groups A, C, and G isolated from blood cultures between January 1, 2007 and May 1, 2012 were included in the study. Isolates were identified to the species level using a combination of phenotypic characteristics and 16s rRNA sequencing. The isolates were subjected to MALDI-ToF analysis. We used a two-stage approach starting with the direct method. If no valid result was obtained we proceeded to an extraction protocol. Scores above 2 were considered valid identification at the species level. A total of 97 Streptococcus pyogenes, 133 Streptococcus dysgalactiae, and 2 Streptococcus canis isolates were tested; 94%, 66%, and 100% of S. pyogenes, S. dysgalactiae, and S. canis, respectively, were correctly identified by MALDI-ToF. In most instances when the isolates were not identified by MALDI-ToF this was because MALDI-ToF was unable to differentiate between S. pyogenes and S. dysgalactiae. By removing two S. pyogenes reference spectra from the MALDI-ToF database the proportion of correctly identified isolates increased to 96% overall. MALDI-ToF is a promising method for discriminating between S. dysgalactiae, S. canis, and S. equi, although more strains need to be tested to clarify this.

  16. Development of a rapid and simplified protocol for direct bacterial identification from positive blood cultures by using matrix assisted laser desorption ionization time-of- flight mass spectrometry.

    Jakovljev, Aleksandra; Bergh, Kåre

    2015-11-06

    Bloodstream infections represent serious conditions carrying a high mortality and morbidity rate. Rapid identification of microorganisms and prompt institution of adequate antimicrobial therapy is of utmost importance for a successful outcome. Aiming at the development of a rapid, simplified and efficient protocol, we developed and compared two in-house preparatory methods for the direct identification of bacteria from positive blood culture flasks (BD BACTEC FX system) by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF MS). Both methods employed saponin and distilled water for erythrocyte lysis. In method A the cellular pellet was overlaid with formic acid on the MALDI TOF target plate for protein extraction, whereas in method B the pellet was exposed to formic acid followed by acetonitrile prior to placing on the target plate. Best results were obtained by method A. Direct identification was achieved for 81.9 % and 65.8 % (50.3 % and 26.2 % with scores >2.0) of organisms by method A and method B, respectively. Overall concordance with final identification was 100 % to genus and 97.9 % to species level. By applying a lower cut-off score value, the levels of identification obtained by method A and method B increased to 89.3 % and 77.8 % of organisms (81.9 % and 65.8 % identified with scores >1.7), respectively. Using the lowered score criteria, concordance with final results was obtained for 99.3 % of genus and 96.6 % of species identifications. The reliability of results, rapid performance (approximately 25 min) and applicability of in-house method A have contributed to implementation of this robust and cost-effective method in our laboratory.

  17. Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry.

    La Scola, Bernard; Raoult, Didier

    2009-11-25

    With long delays observed between sampling and availability of results, the usefulness of blood cultures in the context of emergency infectious diseases has recently been questioned. Among methods that allow quicker bacterial identification from growing colonies, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry was demonstrated to accurately identify bacteria routinely isolated in a clinical biology laboratory. In order to speed up the identification process, in the present work we attempted bacterial identification directly from blood culture bottles detected positive by the automate. We prospectively analysed routine MALDI-TOF identification of bacteria detected in blood culture by two different protocols involving successive centrifugations and then lysis by trifluoroacetic acid or formic acid. Of the 562 blood culture broths detected as positive by the automate and containing one bacterial species, 370 (66%) were correctly identified. Changing the protocol from trifluoroacetic acid to formic acid improved identification of Staphylococci, and overall correct identification increased from 59% to 76%. Lack of identification was observed mostly with viridans streptococci, and only one false positive was observed. In the 22 positive blood culture broths that contained two or more different species, only one of the species was identified in 18 samples, no species were identified in two samples and false species identifications were obtained in two cases. The positive predictive value of bacterial identification using this procedure was 99.2%. MALDI-TOF MS is an efficient method for direct routine identification of bacterial isolates in blood culture, with the exception of polymicrobial samples and viridans streptococci. It may replace routine identification performed on colonies, provided improvement for the specificity of blood culture broths growing viridans streptococci is obtained in the near future.

  18. The use of Gram stain and matrix-assisted laser desorption ionization time-of-flight mass spectrometry on positive blood culture: synergy between new and old technology.

    Fuglsang-Damgaard, David; Nielsen, Camilla Houlberg; Mandrup, Elisabeth; Fuursted, Kurt

    2011-10-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is promising as an alternative to more costly and cumbersome methods for direct identifications in blood cultures. We wanted to evaluate a simplified pre-treatment method for using MALDI-TOF-MS directly on positive blood cultures using BacT/Alert blood culture system, and to test an algorithm combining the result of the initial microscopy with the result suggested by MALDI-TOF-MS. Using the recommended cut-off score of 1.7 the best results were obtained among Gram-negative rods with correct identifications in 91% of Enterobacteriaceae, 83% in aerobic/non-fermentative Gram-negative rods, whereas results were more modest among Gram-positive cocci with correct identifications in 52% of Staphylococci, 54% in Enterococci and only 20% in Streptococci. Combining the results of Gram stain with the top reports by MALDI-TOF-MS, increased the sensitivity from 91% to 93% in the score range from 1.5 to 1.7 and from 48% to 85% in the score range from 1.3 to 1.5. Thus, using this strategy and accepting a cut-off at 1.3 instead of the suggested 1.7, overall sensitivity could be increased from 88.1% to 96.3%. MALDI-TOF-MS is an efficient method for direct routine identification of bacterial isolates in blood culture, especially when combined with the result of the Gram stain. © 2011 The Authors. APMIS © 2011 APMIS.

  19. Diagnostic performance of matrix-assisted laser desorption ionisation time-of-flight mass spectrometry in blood bacterial infections: a systematic review and meta-analysis.

    Scott, Jamie S; Sterling, Sarah A; To, Harrison; Seals, Samantha R; Jones, Alan E

    2016-07-01

    Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) has shown promise in decreasing time to identification of causative organisms compared to traditional methods; however, the utility of MALDI-TOF MS in a heterogeneous clinical setting is uncertain. To perform a systematic review on the operational performance of the Bruker MALDI-TOF MS system and evaluate published cut-off values compared to traditional blood cultures. A comprehensive literature search was performed. Studies were included if they performed direct MALDI-TOF MS analysis of blood culture specimens in human patients with suspected bacterial infections using the Bruker Biotyper software. Sensitivities and specificities of the combined studies were estimated using a hierarchical random effects linear model (REML) incorporating cut-off scores of ≥1.7 and ≥2.0. Fifty publications were identified, with 11 studies included after final review. The estimated sensitivity utilising a cut-off of ≥2.0 from the combined studies was 74.6% (95% CI = 67.9-89.3%), with an estimated specificity of 88.0% (95% CI = 74.8-94.7%). When assessing a cut-off of ≥1.7, the combined sensitivity increases to 92.8% (95% CI = 87.4-96.0%), but the estimated specificity decreased to 81.2% (95% CI = 61.9-96.6%). In this analysis, MALDI-TOF MS showed acceptable sensitivity and specificity in bacterial speciation with the current recommended cut-off point compared to blood cultures; however, lowering the cut-off point from ≥2.0 to ≥1.7 would increase the sensitivity of the test without significant detrimental effect on the specificity, which could improve clinician confidence in their results.

  20. Identification of bacteria in blood culture broths using matrix-assisted laser desorption-ionization Sepsityper™ and time of flight mass spectrometry.

    Jen Kok

    Full Text Available Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS is a novel method for the direct identification of bacteria from blood culture broths. We evaluate for the first time, the performance of the MALDI Sepsityper™ Kit and MS for the identification of bacteria compared to standard phenotypic methods using the manufacturer's specified bacterial identification criteria (spectral scores ≥1.700-1.999 and ≥2.000 indicated identification to genus and species level, respectively. Five hundred and seven positive blood culture broths were prospectively examined, of which 379 (74.8%; 358 monomicrobial, 21 polymicrobial were identified by MALDI-TOF MS; 195 (100% and 132 (67.7% of 195 gram-positive; and 163 (100% and 149 (91.4% of 163 gram-negative organisms from monomicrobial blood cultures were correctly identified to genus and species level, respectively. Spectral scores <1.700 (no identification were obtained in 128/507 (25.2% positive blood culture broths, including 31.6% and 32.3% of gram-positive and polymicrobial blood cultures, respectively. Significantly more gram-negative organisms were identified compared to gram-positive organisms at species level (p<0.0001. Five blood cultures were misidentified, but at species level only; including four monomicrobial blood cultures with Streptococcus oralis/mitis that were misidentified as Streptococcus pneumoniae. Positive predictive values for the direct identification of both gram-positive and gram-negative bacteria from monomicrobial blood culture broths to genus level were 100%. A diagnostic algorithm for positive blood culture broths that incorporates gram staining and MALDI-TOF MS should identify the majority of pathogens, particularly to genus level.

  1. Matrix-assisted laser desorption ionization time of flight mass spectrometry and diagnostic testing for prosthetic joint infection in the clinical microbiology laboratory.

    Peel, Trisha N; Cole, Nicolynn C; Dylla, Brenda L; Patel, Robin

    2015-03-01

    Identification of pathogen(s) associated with prosthetic joint infection (PJI) is critical for patient management. Historically, many laboratories have not routinely identified organisms such as coagulase-negative staphylococci to the species level. The advent of matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) has enhanced clinical laboratory capacity for accurate species-level identification. The aim of this study was to describe the species-level identification of microorganisms isolated from periprosthetic tissue and fluid specimens using MALDI-TOF MS alongside other rapid identification tests in a clinical microbiology laboratory. Results of rapid identification of bacteria isolated from periprosthetic joint fluid and/or tissue specimens were correlated with clinical findings at Mayo Clinic, Rochester, Minnesota, between May 2012 and May 2013. There were 178 PJI and 82 aseptic failure (AF) cases analyzed, yielding 770 organisms (median, 3/subject; range, 1-19/subject). MALDI-TOF MS was employed for the identification of 455 organisms (59%) in 197 subjects (123 PJIs and 74 AFs), with 89% identified to the species level using this technique. Gram-positive bacteria accounted for 68% and 93% of isolates in PJI and AF, respectively. However, the profile of species associated with infection compared to specimen contamination differed. Staphylococcus aureus and Staphylococcus caprae were always associated with infection, Staphylococcus epidermidis and Staphylococcus lugdunensis were equally likely to be a pathogen or a contaminant, whereas the other coagulase-negative staphylococci were more frequently contaminants. Most streptococcal and Corynebacterium isolates were pathogens. The likelihood that an organism was a pathogen or contaminant differed with the prosthetic joint location, particularly in the case of Propionibacterium acnes. MALDI-TOF MS is a valuable tool for the identification of bacteria isolated from patients

  2. Ceria nanocubic-ultrasonication assisted dispersive liquid-liquid microextraction coupled with matrix assisted laser desorption/ionization mass spectrometry for pathogenic bacteria analysis.

    Abdelhamid, Hani Nasser; Bhaisare, Mukesh L; Wu, Hui-Fen

    2014-03-01

    A new ceria (CeO2) nanocubic modified surfactant is used as the basis of a novel nano-based microextraction technique for highly sensitive detection of pathogenic bacteria (Pseudomonas aeruginosa and Staphylococcus aureus). The technique uses ultrasound enhanced surfactant-assisted dispersive liquid-liquid microextraction (UESA-DLLME) with and without ceria (CeO2) followed by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). In order to achieve high separation efficiency, we investigated the influential parameters, including extraction time of ultrasonication, type and volume of the extraction solvent and surfactant. Among various surfactants, the cationic surfactants can selectively offer better extraction efficiency on bacteria analysis than that of the anionic surfactants due to the negative charges of bacteria cell membranes. Extractions of the bacteria lysate from aqueous samples via UESA-DLLME-MALDI-MS were successfully achieved by using cetyltrimethyl ammonium bromide (CTAB, 10.0 µL, 1.0×10(-3) M) as surfactants in chlorobenzene (10.0 µL) and chloroform (10.0 µL) as the optimal extracting solvent for P. aeruginosa and S. aureus, respectively. Ceria nanocubic was synthesized, and functionalized with CTAB (CeO2@CTAB) and then characterized using transmission electron microscopy (TEM) and optical spectroscopy (UV and FTIR). CeO2@CTAB demonstrates high extraction efficiency, improve peaks ionization, and enhance resolution. The prime reasons for these improvements are due to the large surface area of nanoparticles, and its absorption that coincides with the wavelength of MALDI laser (337 nm, N2 laser). CeO2@CTAB-based microextraction offers lowest detectable concentrations tenfold lower than that of without nanoceria. The present approach has been successfully applied to detect pathogenic bacteria at low concentrations of 10(4)-10(5) cfu/mL (without ceria) and at 10(3)-10(4) cfu/mL (with ceria) from bacteria suspensions. Finally, the

  3. A SIMPLE AND RAPID MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY METHOD TO SCREEN FISH PLASMA SAMPLES FOR ESTROGEN-RESPONSIVE BIOMARKERS

    In this study, we describe and evaluate the performance of a simple and rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers using matrix assisted laster desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) couopled with a short...

  4. High power ultrashort pulse lasers

    Perry, M.D.

    1994-01-01

    Small scale terawatt and soon even petawatt (1000 terawatt) class laser systems are made possible by application of the chirped-pulse amplification technique to solid-state lasers combined with the availability of broad bandwidth materials. These lasers make possible a new class of high gradient accelerators based on the large electric fields associated with intense laser-plasma interactions or from the intense laser field directly. Here, we concentrate on the laser technology to produce these intense pulses. Application of the smallest of these systems to the production of high brightness electron sources is also introduced

  5. Pulsed laser thin film growth of di-octyl substituted polyfluorene and its co-polymers

    Gupta, R.K.; Ghosh, K.; Kahol, P.K. [Department of Physics, Astronomy and Materials Science, Missouri State University, Springfield, MO 65897 (United States); Yoon, J. [Department of Physics and Astronomy, University of Missouri, Columbia, MO 65211 (United States); Guha, S. [Department of Physics and Astronomy, University of Missouri, Columbia, MO 65211 (United States)], E-mail: guhas@missouri.edu

    2008-08-30

    Matrix-assisted pulsed laser deposition (PLD) allows a controlled layer-by-layer growth of polymer films. Di-octyl substituted polyfluorene (PF8) and its copolymers were deposited as thin films using matrix-assisted PLD by employing a KrF excimer laser with a fluence of 125 mJ/pulses. The optical and structural properties of these films are compared with spincoated films via Raman spectroscopy, absorption and photoluminescence. The Raman spectra of both PLD and spincoated films are similar indicating that the polymer films deposited via PLD maintain their molecular structure. Both the spincoated and the PLD grown PF8 films that were cast from toluene show the presence of the {beta} phase. Benzothiadiazole substituted PF8 (F8BT) and butyl phenyl-substituted PF8 (PFB) PLD grown films show a slightly broader emission compared to the spincoated films, which is attributed to an enhanced intermolecular interaction in the PLD grown thin films.

  6. Pulsed laser thin film growth of di-octyl substituted polyfluorene and its co-polymers

    Gupta, R.K.; Ghosh, K.; Kahol, P.K.; Yoon, J.; Guha, S.

    2008-01-01

    Matrix-assisted pulsed laser deposition (PLD) allows a controlled layer-by-layer growth of polymer films. Di-octyl substituted polyfluorene (PF8) and its copolymers were deposited as thin films using matrix-assisted PLD by employing a KrF excimer laser with a fluence of 125 mJ/pulses. The optical and structural properties of these films are compared with spincoated films via Raman spectroscopy, absorption and photoluminescence. The Raman spectra of both PLD and spincoated films are similar indicating that the polymer films deposited via PLD maintain their molecular structure. Both the spincoated and the PLD grown PF8 films that were cast from toluene show the presence of the β phase. Benzothiadiazole substituted PF8 (F8BT) and butyl phenyl-substituted PF8 (PFB) PLD grown films show a slightly broader emission compared to the spincoated films, which is attributed to an enhanced intermolecular interaction in the PLD grown thin films

  7. Pulsed laser thin film growth of di-octyl substituted polyfluorene and its co-polymers

    Gupta, R. K.; Ghosh, K.; Kahol, P. K.; Yoon, J.; Guha, S.

    2008-08-01

    Matrix-assisted pulsed laser deposition (PLD) allows a controlled layer-by-layer growth of polymer films. Di-octyl substituted polyfluorene (PF8) and its copolymers were deposited as thin films using matrix-assisted PLD by employing a KrF excimer laser with a fluence of 125 mJ/pulses. The optical and structural properties of these films are compared with spincoated films via Raman spectroscopy, absorption and photoluminescence. The Raman spectra of both PLD and spincoated films are similar indicating that the polymer films deposited via PLD maintain their molecular structure. Both the spincoated and the PLD grown PF8 films that were cast from toluene show the presence of the β phase. Benzothiadiazole substituted PF8 (F8BT) and butyl phenyl-substituted PF8 (PFB) PLD grown films show a slightly broader emission compared to the spincoated films, which is attributed to an enhanced intermolecular interaction in the PLD grown thin films.

  8. Typing of Ochrobactrum anthropi clinical isolates using automated repetitive extragenic palindromic-polymerase chain reaction DNA fingerprinting and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

    Quirino, Angela; Pulcrano, Giovanna; Rametti, Linda; Puccio, Rossana; Marascio, Nadia; Catania, Maria Rosaria; Matera, Giovanni; Liberto, Maria Carla; Focà, Alfredo

    2014-03-22

    Ochrobactrum anthropi (O. anthropi), is a non-fermenting gram-negative bacillus usually found in the environment. Nevertheless, during the past decade it has been identified as pathogenic to immunocompromised patients. In this study, we assessed the usefulness of the automated repetitive extragenic palindromic-polymerase chain reaction (rep-PCR-based DiversiLab™ system, bioMèrieux, France) and of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF MS) for typing of twentythree O. anthropi clinical isolates that we found over a four-months period (from April 2011 to August 2011) in bacteriemic patients admitted in the same operative unit of our hospital. Pulsed-field gel electrophoresis (PFGE), commonly accepted as the gold standard technique for typing, was also used. Analysis was carried out using the Pearson correlation coefficient to determine the distance matrice and the unweighted pair group method with arithmetic mean (UPGMA) to generate dendogram. Rep-PCR analysis identified four different patterns: three that clustered together with 97% or more pattern similarity, and one whose members showed < 95% pattern similarity. Interestingly, strains isolated later (from 11/06/2011 to 24/08/2011) displayed a pattern with 99% similarity. MALDI-TOF MS evaluation clustered the twentythree strains of O. anthropi into a single group containing four distinct subgroups, each comprising the majority of strains clustering below 5 distance levels, indicating a high similarity between the isolates. Our results indicate that these isolates are clonally-related and the methods used afforded a valuable contribution to the epidemiology, prevention and control of the infections caused by this pathogen.

  9. Direct detection of carbapenemase-associated proteins of Acinetobacter baumannii using nanodiamonds coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Chang, Kai-Chih; Chung, Chin-Yi; Yeh, Chen-Hsing; Hsu, Kuo-Hsiu; Chin, Ya-Ching; Huang, Sin-Siang; Liu, Bo-Rong; Chen, Hsi-An; Hu, Anren; Soo, Po-Chi; Peng, Wen-Ping

    2018-04-01

    The appearance and spread of carbapenem-resistant Acinetobacter baumannii (CRAB) pose a challenge for optimization of antibiotic therapies and outbreak preventions. The carbapenemase production can be detected through culture-based methods (e.g. Modified Hodge Test-MHT) and DNA based methods (e.g. Polymerase Chain Reaction-PCR). The culture-based methods are time-consuming, whereas those of PCR assays need only a few hours but due to its specificity, can only detect known genetic targets encoding carbapenem-resistance genes. Therefore, new approaches to detect carbapenemase-producing A. baumannii are of great importance. Here, we have developed a rapid and novel method using detonation nanodiamonds (DNDs) as a platform for concentration and extraction of A. baumannii carbapenemase-associated proteins prior to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF-MS) analysis. To concentrate and extract the A. baumannii carbapenemase-associated proteins, we tested several protein precipitation conditions and found a 0.5% trifluoroacetic acid (TFA) solution within the bacterial suspension could result in strong ion signals with DNDs. A total of 66 A. baumannii clinical-isolates including 51 carbapenem-resistant strains and 15 carbapenem-susceptible strains were tested. Our result showed that among the 51 carbapenem-resistant strains 49 strains had a signal at m/z ~40,279 (±87); among the 15 carbapenem-susceptible strains, 4 strains showed a signal at m/z ~40,279. With on-diamond digestion, we confirmed that the captured protein at m/z ~40,279 was related to ADC family extended-spectrum class C beta-lactamase, from A. baumannii. Using this ADC family protein as a biomarker (m/z ~ 40,279) for carbapenem susceptibility testing of A. baumannii, the sensitivity and the specificity could reach 96% and 73% as compared to traditional imipenem susceptibility testing (MIC results). However, the sensitivity and specificity of this method

  10. Identification of clinical isolates of Aspergillus, including cryptic species, by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

    Vidal-Acuña, M Reyes; Ruiz-Pérez de Pipaón, Maite; Torres-Sánchez, María José; Aznar, Javier

    2017-12-08

    An expanded library of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been constructed using the spectra generated from 42 clinical isolates and 11 reference strains, including 23 different species from 8 sections (16 cryptic plus 7 noncryptic species). Out of a total of 379 strains of Aspergillus isolated from clinical samples, 179 strains were selected to be identified by sequencing of beta-tubulin or calmodulin genes. Protein spectra of 53 strains, cultured in liquid medium, were used to construct an in-house reference database in the MALDI-TOF MS. One hundred ninety strains (179 clinical isolates previously identified by sequencing and the 11 reference strains), cultured on solid medium, were blindy analyzed by the MALDI-TOF MS technology to validate the generated in-house reference database. A 100% correlation was obtained with both identification methods, gene sequencing and MALDI-TOF MS, and no discordant identification was obtained. The HUVR database provided species level (score of ≥2.0) identification in 165 isolates (86.84%) and for the remaining 25 (13.16%) a genus level identification (score between 1.7 and 2.0) was obtained. The routine MALDI-TOF MS analysis with the new database, was then challenged with 200 Aspergillus clinical isolates grown on solid medium in a prospective evaluation. A species identification was obtained in 191 strains (95.5%), and only nine strains (4.5%) could not be identified at the species level. Among the 200 strains, A. tubingensis was the only cryptic species identified. We demonstrated the feasibility and usefulness of the new HUVR database in MALDI-TOF MS by the use of a standardized procedure for the identification of Aspergillus clinical isolates, including cryptic species, grown either on solid or liquid media. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For

  11. Exploration of polyamide structure-property relationships by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Barrère, Caroline; Rejaibi, Majed; Curat, Aurélien; Hubert-Roux, Marie; Lavanant, Hélène; Afonso, Carlos; Kebir, Nasreddine; Desilles, Nicolas; Lecamp, Laurence; Burel, Fabrice; Loutelier-Bourhis, Corinne

    2014-08-15

    Polyamides (PA) are among the most used classes of polymers because of their attractive properties. Depending on the nature and proportion of the co-monomers used for their synthesis, they can exhibit a very large range of melting temperatures (Tm ). This study aims at the correlation of data from mass spectrometry (MS) with differential scanning calorimetry (DSC) and X-ray diffraction analyses to relate molecular structure to physical properties such as melting temperature, enthalpy change and crystallinity rate. Six different PA copolymers with molecular weights around 3500 g mol(-1) were synthesized with varying proportions of different co-monomers (amino-acid AB/di-amine AA/di-acid BB). Their melting temperature, enthalpy change and crystallinity rate were measured by DSC and X-ray diffraction. Their structural characterization was carried out by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Because of the poor solubility of PA, a solvent-free sample preparation strategy was used with 2,5-dihydroxybenzoic acid (2,5-DHB) as the matrix and sodium iodide as the cationizing agent. The different proportions of the repeating unit types led to the formation of PA with melting temperatures ranging from 115°C to 185°C. The structural characterization of these samples by MALDI-TOF-MS revealed a collection of different ion distributions with different sequences of repeating units (AA, BB; AB/AA, BB and AB) in different proportions according to the mixture of monomers used in the synthesis. The relative intensities of these ion distributions were related to sample complexity and structure. They were correlated to DSC and X-ray results, to explain the observed physical properties. The structural information obtained by MALDI-TOF-MS provided a better understanding of the variation of the PA melting temperature and established a structure-properties relationship. This work will allow future PA designs to be monitored. Copyright

  12. Weak cation exchange magnetic beads coupled with matrix-assisted laser desorption ionization-time of flight-mass spectrometry in screening serum protein markers in osteopenia.

    He, Wei-Tao; Liang, Bo-Cheng; Shi, Zhen-Yu; Li, Xu-Yun; Li, Chun-Wen; Shi, Xiao-Lin

    2016-01-01

    The present study aimed at investigating the weak cation magnetic separation technology and matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) in screening serum protein markers of osteopenia from ten postmenopausal women and ten postmenopausal women without osteopenia as control group, to find a new method for screening biomarkers and establishing a diagnostic model for primary type I osteoporosis. Serum samples were collected from postmenopausal women with osteopenia and postmenopausal women with normal bone mass. Proteins were extracted from serum samples by weak cation exchange magnetic beads technology, and mass spectra acquisition was done by MALDI-TOF-MS. The visualization and comparison of data sets, statistical peak evaluation, model recognition, and discovery of biomarker candidates were handled by the proteinchip data analysis system software(ZJU-PDAS). The diagnostic models were established using genetic arithmetic based support vector machine (SVM). The SVM result with the highest Youden Index was selected as the model. Combinatorial Peaks having the highest accuracy in distinguishing different samples were selected as potential biomarker. From the two group serum samples, a total of 133 differential features were selected. Ten features with significant intensity differences were screened. In the pair-wise comparisons, processing of MALDI-TOF spectra resulted in the identification of ten differential features between postmenopausal women with osteopenia and postmenopausal women with normal bone mass. The difference of features by Youden index showed that the highest features had a mass to charge ratio of 1699 and 3038 Da. A diagnosis model was established with these two peaks as the candidate marker, and the specificity of the model is 100 %, the sensitivity was 90 % by leave-one-out cross validation test. The two groups of specimens in SVM results on the scatter plot could be clearly distinguished. The peak

  13. Direct analysis and identification of pathogenic Lichtheimia species by matrix-assisted laser desorption ionization-time of flight analyzer-mediated mass spectrometry.

    Schrödl, Wieland; Heydel, Tilo; Schwartze, Volker U; Hoffmann, Kerstin; Grosse-Herrenthey, Anke; Walther, Grit; Alastruey-Izquierdo, Ana; Rodriguez-Tudela, Juan Luis; Olias, Philipp; Jacobsen, Ilse D; de Hoog, G Sybren; Voigt, Kerstin

    2012-02-01

    Zygomycetes of the order Mucorales can cause life-threatening infections in humans. These mucormycoses are emerging and associated with a rapid tissue destruction and high mortality. The resistance of Mucorales to antimycotic substances varies between and within clinically important genera such as Mucor, Rhizopus, and Lichtheimia. Thus, an accurate diagnosis before onset of antimycotic therapy is recommended. Matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) is a potentially powerful tool to rapidly identify infectious agents on the species level. We investigated the potential of MALDI-TOF MS to differentiate Lichtheimia species, one of the most important agents of mucormycoses. Using the Bruker Daltonics FlexAnalysis (version 3.0) software package, a spectral database library with m/z ratios of 2,000 to 20,000 Da was created for 19 type and reference strains of clinically relevant Zygomycetes of the order Mucorales (12 species in 7 genera). The database was tested for accuracy by use of 34 clinical and environmental isolates of Lichtheimia comprising a total of five species. Our data demonstrate that MALDI-TOF MS can be used to clearly discriminate Lichtheimia species from other pathogenic species of the Mucorales. Furthermore, the method is suitable to discriminate species within the genus. The reliability and robustness of the MALDI-TOF-based identification are evidenced by high score values (above 2.3) for the designation to a certain species and by moderate score values (below 2.0) for the discrimination between clinically relevant (Lichtheimia corymbifera, L. ramosa, and L. ornata) and irrelevant (L. hyalospora and L. sphaerocystis) species. In total, all 34 strains were unequivocally identified by MALDI-TOF MS with score values of >1.8 down to the generic level, 32 out of 34 of the Lichtheimia isolates (except CNM-CM 5399 and FSU 10566) were identified accurately with score values of >2 (probable species

  14. Direct Analysis and Identification of Pathogenic Lichtheimia Species by Matrix-Assisted Laser Desorption Ionization–Time of Flight Analyzer-Mediated Mass Spectrometry

    Schrödl, Wieland; Heydel, Tilo; Schwartze, Volker U.; Hoffmann, Kerstin; Große-Herrenthey, Anke; Walther, Grit; Alastruey-Izquierdo, Ana; Rodriguez-Tudela, Juan Luis; Olias, Philipp; Jacobsen, Ilse D.; de Hoog, G. Sybren

    2012-01-01

    Zygomycetes of the order Mucorales can cause life-threatening infections in humans. These mucormycoses are emerging and associated with a rapid tissue destruction and high mortality. The resistance of Mucorales to antimycotic substances varies between and within clinically important genera such as Mucor, Rhizopus, and Lichtheimia. Thus, an accurate diagnosis before onset of antimycotic therapy is recommended. Matrix-assisted laser desorption ionization (MALDI)–time of flight (TOF) mass spectrometry (MS) is a potentially powerful tool to rapidly identify infectious agents on the species level. We investigated the potential of MALDI-TOF MS to differentiate Lichtheimia species, one of the most important agents of mucormycoses. Using the Bruker Daltonics FlexAnalysis (version 3.0) software package, a spectral database library with m/z ratios of 2,000 to 20,000 Da was created for 19 type and reference strains of clinically relevant Zygomycetes of the order Mucorales (12 species in 7 genera). The database was tested for accuracy by use of 34 clinical and environmental isolates of Lichtheimia comprising a total of five species. Our data demonstrate that MALDI-TOF MS can be used to clearly discriminate Lichtheimia species from other pathogenic species of the Mucorales. Furthermore, the method is suitable to discriminate species within the genus. The reliability and robustness of the MALDI-TOF-based identification are evidenced by high score values (above 2.3) for the designation to a certain species and by moderate score values (below 2.0) for the discrimination between clinically relevant (Lichtheimia corymbifera, L. ramosa, and L. ornata) and irrelevant (L. hyalospora and L. sphaerocystis) species. In total, all 34 strains were unequivocally identified by MALDI-TOF MS with score values of >1.8 down to the generic level, 32 out of 34 of the Lichtheimia isolates (except CNM-CM 5399 and FSU 10566) were identified accurately with score values of >2 (probable species

  15. Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach.

    Klein, Sabrina; Zimmermann, Stefan; Köhler, Christine; Mischnik, Alexander; Alle, Werner; Bode, Konrad A

    2012-03-01

    Sepsis is a major cause of mortality in hospitalized patients worldwide, with lethality rates ranging from 30 to 70 %. Sepsis is caused by a variety of different pathogens, and rapid diagnosis is of outstanding importance, as early and adequate antimicrobial therapy correlates with positive clinical outcome. In recent years, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) fingerprinting has become a powerful tool in microbiological diagnostics. The direct identification of micro-organisms in a positive blood culture by MALDI-TOF MS can shorten the diagnostic procedure significantly. Therefore, the aim of the present study was to evaluate whether identification rates could be improved by using the new Sepsityper kit from Bruker Daltonics for direct isolation and identification of bacteria from positive blood cultures by MALDI-TOF MS compared with the use of conventional separator gel columns, and to integrate the MALDI-TOF MS-based identification method into the routine course of blood culture diagnostics in the setting of a microbiological laboratory at a university hospital in Germany. The identification of Gram-negative bacteria by MALDI-TOF MS was significantly better using the Sepsityper kit compared with a separator gel tube-based method (99 and 68 % correct identification, respectively). For Gram-positive bacteria, only 73 % were correctly identified by MALDI-TOF with the Sepsityper kit and 59 % with the separator gel tube assay. A major problem of both methods was the poor identification of Gram-positive grape-like clustered cocci. As differentiation of Staphylococcus aureus from coagulase-negative staphylococci is of clinical importance, a PCR was additionally established that was capable of identifying S. aureus directly from positive blood cultures, thus closing this diagnostic gap. Another benefit of the PCR approach is the possibility of directly detecting the genes responsible for meticillin

  16. A matrix-assisted laser desorption/ionization mass spectroscopy method for the analysis of small molecules by integrating chemical labeling with the supramolecular chemistry of cucurbituril.

    Ding, Jun; Xiao, Hua-Ming; Liu, Simin; Wang, Chang; Liu, Xin; Feng, Yu-Qi

    2018-10-05

    Although several methods have realized the analysis of low molecular weight (LMW) compounds using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) by overcoming the problem of interference with MS signals in the low mass region derived from conventional organic matrices, this emerging field still requires strategies to address the issue of analyzing complex samples containing LMW components in addition to the LMW compounds of interest, and solve the problem of lack of universality. The present study proposes an integrated strategy that combines chemical labeling with the supramolecular chemistry of cucurbit [n]uril (CB [n]) for the MALDI MS analysis of LMW compounds in complex samples. In this strategy, the target LMW compounds are first labeled by introducing a series of bifunctional reagents that selectively react with the target analytes and also form stable inclusion complexes with CB [n]. Then, the labeled products act as guest molecules that readily and selectively form stable inclusion complexes with CB [n]. This strategy relocates the MS signals of the LMW compounds of interest from the low mass region suffering high interference to the high mass region where interference with low mass components is absent. Experimental results demonstrate that a wide range of LMW compounds, including carboxylic acids, aldehydes, amines, thiol, and cis-diols, can be successfully detected using the proposed strategy, and the limits of detection were in the range of 0.01-1.76 nmol/mL. In addition, the high selectivity of the labeling reagents for the target analytes in conjunction with the high selectivity of the binding between the labeled products and CB [n] ensures an absence of signal interference with the non-targeted LMW components of complex samples. Finally, the feasibility of the proposed strategy for complex sample analysis is demonstrated by the accurate and rapid quantitative analysis of aldehydes in saliva and herbal

  17. Laser system using ultra-short laser pulses

    Dantus, Marcos [Okemos, MI; Lozovoy, Vadim V [Okemos, MI; Comstock, Matthew [Milford, MI

    2009-10-27

    A laser system using ultrashort laser pulses is provided. In another aspect of the present invention, the system includes a laser, pulse shaper and detection device. A further aspect of the present invention employs a femtosecond laser and binary pulse shaping (BPS). Still another aspect of the present invention uses a laser beam pulse, a pulse shaper and a SHG crystal.

  18. High-power pulsed lasers

    Holzrichter, J.F.

    1980-01-01

    The ideas that led to the successful construction and operation of large multibeam fusion lasers at the Lawrence Livermore Laboratory are reviewed. These lasers are based on the use of Nd:glass laser materials. However, most of the concepts are applicable to any laser being designed for fusion experimentation. This report is a summary of lectures given by the author at the 20th Scottish University Summer School in Physics, on Laser Plasma Interaction. This report includes basic concepts of the laser plasma system, a discussion of lasers that are useful for short-pulse, high-power operation, laser design constraints, optical diagnostics, and system organization

  19. Rapid Detection and Identification of Candidemia by Direct Blood Culturing on Solid Medium by Use of Lysis-Centrifugation Method Combined with Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (MALDI-TOF MS)

    Idelevich, Evgeny A.; Grünastel, Barbara

    2016-01-01

    ABSTRACT Candida sepsis is a life-threatening condition with increasing prevalence. In this study, direct blood culturing on solid medium using a lysis-centrifugation procedure enabled successful Candida species identification by matrix-assisted laser desorption–ionization time of flight mass spectrometry on average 3.8 h (Sabouraud agar) or 7.4 h (chocolate agar) before the positivity signal for control samples in Bactec mycosis-IC/F or Bactec Plus aerobic/F bottles, respectively. Direct culturing on solid medium accelerated candidemia diagnostics compared to that with automated broth-based systems. PMID:27795344

  20. Rapid Detection and Identification of Candidemia by Direct Blood Culturing on Solid Medium by Use of Lysis-Centrifugation Method Combined with Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS).

    Idelevich, Evgeny A; Grünastel, Barbara; Becker, Karsten

    2017-01-01

    Candida sepsis is a life-threatening condition with increasing prevalence. In this study, direct blood culturing on solid medium using a lysis-centrifugation procedure enabled successful Candida species identification by matrix-assisted laser desorption-ionization time of flight mass spectrometry on average 3.8 h (Sabouraud agar) or 7.4 h (chocolate agar) before the positivity signal for control samples in Bactec mycosis-IC/F or Bactec Plus aerobic/F bottles, respectively. Direct culturing on solid medium accelerated candidemia diagnostics compared to that with automated broth-based systems. Copyright © 2016 American Society for Microbiology.

  1. Matrix-assisted laser desorption/ionization time of flight mass spectrometry for comprehensive indexing of East African ixodid tick species.

    Rothen, Julian; Githaka, Naftaly; Kanduma, Esther G; Olds, Cassandra; Pflüger, Valentin; Mwaura, Stephen; Bishop, Richard P; Daubenberger, Claudia

    2016-03-15

    The tick population of Africa includes several important genera belonging to the family Ixodidae. Many of these ticks are vectors of protozoan and rickettsial pathogens including Theileria parva that causes East Coast fever, a debilitating cattle disease endemic to eastern, central and southern Africa. Effective surveillance of tick-borne pathogens depends on accurate identification and mapping of their tick vectors. A simple and reproducible technique for rapid and reliable differentiation of large numbers of closely related field-collected ticks, which are often difficult and tedious to discriminate purely by morphology, will be an essential component of this strategy. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is increasingly becoming a useful tool in arthropod identification and has the potential to overcome the limitations of classical morphology-based species identification. In this study, we applied MALDI-TOF MS to a collection of laboratory and field ticks found in Eastern Africa. The objective was to determine the utility of this proteomic tool for reliable species identification of closely related afrotropical ticks. A total of 398 ixodid ticks from laboratory maintained colonies, extracted from the hides of animals or systematically collected from vegetation in Kenya, Sudan and Zimbabwe were analyzed in the present investigation. The cytochrome c oxidase I (COI) genes from 33 specimens were sequenced to confirm the tentatively assigned specimen taxa identity on the basis of morphological analyses. Subsequently, the legs of ticks were homogenized and analyzed by MALDI-TOF MS. A collection of reference mass spectra, based on the mass profiles of four individual ticks per species, was developed and deposited in the spectral database SARAMIS™. The ability of these superspectra (SSp.) to identify and reliably validate a set of ticks was demonstrated using the remaining individual 333 ticks. Ultimately, ten

  2. [Identification of mycobacteria by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry--using reference strains and clinical isolates of Mycobacterium].

    Niitsuma, Katsunao; Saito, Miwako; Koshiba, Shizuko; Kaneko, Michiyo

    2014-05-01

    Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) method is being played an important role for the inspection of clinical microorganism as a rapid and the price reduction. Mass spectra obtained by measuring become points of identification whether the peak pattern match any species mass spectral pattern. We currently use MALDI-TOF MS for rapid and accurate diagnosis of inactivated reference and clinical isolates of Mycobacterium because of the improved pretreatment techniques compared with former inspection methods that pose a higher risk of infection to the operator. The identification matching rate of score value (SV) peak pattern spectra was compared with that of conventional methods such as strain diffusion/amplification. Also, cultures were examined after a fixed number of days. Compared with the initial inspection technique, the pretreatment stage of current MALDI-TOF MS inspection techniques can improve the analysis of inactivated acid-fast bacteria that are often used as inspection criteria strains of clinical isolates. Next, we compared the concordance rate for identification between MALDI-TOF MS and conventional methods such as diffusion/amplification by comparison of peak pattern spectra and evaluated SV spectra to identify differences in the culture media after the retention period. In examination of 158 strains of clinical isolated Mycobacterium tuberculosis complex (MTC), the identification coincidence rate in the genus level in a matching pattern was 99.4%, when the species level was included 94.9%. About 37 strains of nontuberculous mycobacteria (NTM), the identification coincidence rate in the genus level was 94.6%. M. bovis BCG (Tokyo strain) in the reference strain was judged by the matching pattern to be MTC, and it suggested that they are M. tuberculosis and affinity species with high DNA homology. Nontuberculous mycobacterial M. gordonae strain JATA 33-01 shared peak pattern spectra, excluding the

  3. Matrix Assisted and/or Laser Desorption Ionization Quadrupole Ion Trap Time-of-Flight Mass Spectrometry of WO3 Clusters Formation in Gas Phase. Nanodiamonds, Fullerene, and Graphene Oxide Matrices

    Ausekar, Mayuri Vilas; Mawale, Ravi Madhukar; Pazdera, Pavel; Havel, Josef

    2018-03-01

    The formation of W x O y +●/-● clusters in the gas phase was studied by laser desorption ionization (LDI) and matrix assisted laser desorption ionization (MALDI) of solid WO3. LDI produced (WO3) n + ●/- ● ( n = 1-7) clusters. In MALDI, when using nano-diamonds (NDs), graphene oxide (GO), or fullerene (C60) matrices, higher mass clusters were generated. In addition to (WO3) n -● clusters, oxygen-rich or -deficient species were found in both LDI and MALDI (with the total number of clusters exceeding one hundred ≈ 137). This is the first time that such matrices have been used for the generation of(WO3) n + ●/-● clusters in the gas phase, while new high mass clusters (WO3) n -● ( n = 12-19) were also detected. [Figure not available: see fulltext.

  4. Coaxial short pulsed laser

    Nelson, M.A.; Davies, T.J.

    1975-08-01

    This invention relates to a laser system of rugged design suitable for use in a field environment. The laser itself is of coaxial design with a solid potting material filling the space between components. A reservoir is employed to provide a gas lasing medium between an electrode pair, each of which is connected to one of the coaxial conductors. (auth)

  5. Pulsed laser deposition of the lysozyme protein: an unexpected “Inverse MAPLE” process

    Schou, Jørgen; Matei, Andreea; Constantinescu, Catalin

    2012-01-01

    Films of organic materials are commonly deposited by laser assisted methods, such as MAPLE (matrix-assisted pulsed laser evaporation), where a few percent of the film material in the target is protected by a light-absorbing volatile matrix. Another possibility is to irradiate the dry organic...... the ejection and deposition of lysozyme. This can be called an “inverse MAPLE” process, since the ratio of “matrix” to film material in the target is 10:90, which is inverse of the typical MAPLE process where the film material is dissolved in the matrix down to several wt.%. Lysozyme is a well-known protein...

  6. Pulsed chemical laser

    Jacobson, T.V.; Kimbell, G.H.

    1975-01-01

    A hydrogen fluoride laser capable of operating super radiantly and at atmospheric pressure is described. A transverse electrical discharge is utilized to energize the reaction of a hydrogen donor to provide hydrogen fluoride in a metastable energy state which reverts to a stable state by laser action. A large range of hydrogen and fluorine donors is disclosed. A preferred pair of donors is sulphur hexafluoride and propane. Helium is frequently added to the gas mix to act as a buffer. (U.S.)

  7. Matrix-assisted laser desorption/ionization time-of-flight and nano-electrospray ionization ion trap mass spectrometric characterization of 1-cyano-2-substituted-benz[f]isoindole derivatives of peptides for fluorescence detection

    Linnemayr, K; Brückner, A; Körner, R

    1999-01-01

    A series of hexa- to decapeptides (molecular mass range 800-1200) were labeled with naphthalene-2,3-dicarboxaldehyde, which preferentially reacts with the primary amino groups of a peptide. A highly stable peptide conjugate is formed, which allows selective analysis by fluorescence at excitation...... and emission wavelengths of 420 and 490 nm, respectively. After removal of unreacted compounds, the peptide conjugates were characterized by matrix-assisted laser desorption/ionization (MALDI) time-of-flight and nano-electrospray ionization (ESI) ion trap mass spectrometry. They readily form both [M + H]+ ions...... by MALDI and both [M + H]+ and [M + 2H]2+ ions by ESI. Furthermore, the fragmentation behavior of the N-terminally tagged peptides, exhibiting an uncharged N-terminus, was investigated applying post-source decay fragmentation with a curved field reflector and collision-induced dissociation...

  8. A microwave-mediated saponification of galactosylceramide and galactosylceramide I3-sulfate and identification of their lyso-compounds by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Taketomi, T; Hara, A; Uemura, K; Kurahashi, H; Sugiyama, E

    1996-07-16

    Small amounts of galactosylceramide (cerebroside) and galactosylceramide I3-sulfate (sulfatide) obtained from porcine spinal cord and equine kidney were deacylated by a rapid method of microwave-mediated saponification to prepare their lyso-compounds. Mass spectra of their protonated or deprotonated molecular ion peaks were detected by recently developed new technology of a delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometer with reflector detector in positive or negative ion mode. Long chain bases of lysocerebroside and lysosulfatide were different between porcine spinal cord and equine kidney, but similar to each other in the same organ, suggesting their common synthetic pathway. It is noted that the new rapid method can be similarly applied to the deacylation of both cerebroside and sulfatide in contrast to our classical method which was able to be applied to cerebroside, but not to sulfatide.

  9. Proteome-based bacterial identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS): A revolutionary shift in clinical diagnostic microbiology.

    Nomura, Fumio

    2015-06-01

    Rapid and accurate identification of microorganisms, a prerequisite for appropriate patient care and infection control, is a critical function of any clinical microbiology laboratory. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a quick and reliable method for identification of microorganisms, including bacteria, yeast, molds, and mycobacteria. Indeed, there has been a revolutionary shift in clinical diagnostic microbiology. In the present review, the state of the art and advantages of MALDI-TOF MS-based bacterial identification are described. The potential of this innovative technology for use in strain typing and detection of antibiotic resistance is also discussed. This article is part of a Special Issue entitled: Medical Proteomics. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. The sensitivity of direct identification from positive BacT/ALERT™ (bioMérieux) blood culture bottles by matrix-assisted laser desorption ionization time-of-flight mass spectrometry is low.

    Szabados, F; Michels, M; Kaase, M; Gatermann, S

    2011-02-01

    Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been presented as a novel method for the direct identification of bacteria from positive blood culture bottles. The rate of the MALDI TOF MS-based identification in the present study from positive BacT/ALERT (bioMérieux, Marcy l'Etoile, France) blood culture bottles was 30%, which is far below the previously reported sensitivities using the BACTEC (Becton Dickinson, Franklin Lakes, NJ, USA) system. We also found evidence that the Biotyper algorithm did not identify a second pathogen in cases of positive BacT/ALERT blood culture bottles containing two different species. © 2010 The Authors. Journal Compilation © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  11. Rapid method for direct identification of bacteria in urine and blood culture samples by matrix-assisted laser desorption ionization time-of-flight mass spectrometry: intact cell vs. extraction method.

    Ferreira, L; Sánchez-Juanes, F; Muñoz-Bellido, J L; González-Buitrago, J M

    2011-07-01

    Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable technology for the identification of microorganisms with proteomics approaches. Here, we compare an intact cell method and a protein extraction method before application on the MALDI plate for the direct identification of microorganisms in both urine and blood culture samples from clinical microbiology laboratories. The results show that the intact cell method provides excellent results for urine and is a good initial method for blood cultures. The extraction method complements the intact cell method, improving microorganism identification from blood culture. Thus, we consider that MALDI-TOF MS performed directly on urine and blood culture samples, with the protocols that we propose, is a suitable technique for microorganism identification, as compared with the routine methods used in the clinical microbiology laboratory. © 2010 The Authors. Clinical Microbiology and Infection © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  12. Direct detection of the plant pathogens Burkholderia glumae, Burkholderia gladioli pv. gladioli, and Erwinia chrysanthemi pv. zeae in infected rice seedlings using matrix assisted laser desorption/ionization time-of-flight mass spectrometry.

    Kajiwara, Hideyuki

    2016-01-01

    The plant pathogens Burkholderia glumae, Burkholderia gladioli pv. gladioli, and Erwinia chrysanthemi pv. zeae were directly detected in extracts from infected rice seedlings by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This method did not require culturing of the pathogens on artificial medium. In the MALDI-TOF MS analysis, peaks originating from bacteria were found in extracts from infected rice seedlings. The spectral peaks showed significantly high scores, in spite of minor differences in spectra. The spectral peaks originating from host plant tissues did not affect this direct MALDI-TOF MS analysis for the rapid identification of plant pathogens. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Recognition of Streptococcus pseudoporcinus Colonization in Women as a Consequence of Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Group B Streptococcus Identification.

    Suwantarat, Nuntra; Grundy, Maureen; Rubin, Mayer; Harris, Renee; Miller, Jo-Anne; Romagnoli, Mark; Hanlon, Ann; Tekle, Tsigereda; Ellis, Brandon C; Witter, Frank R; Carroll, Karen C

    2015-12-01

    During a 14-month period of using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for group B streptococcus (GBS) identification, we recovered 32 (1%) Streptococcus pseudoporcinus isolates from 3,276 GBS screening cultures from female genital sources (25 isolates from pregnant women and 7 from nonpregnant women). An additional two S. pseudoporcinus isolates were identified from a urine culture and a posthysterectomy wound culture. These isolates were found to cross-react with three different GBS antigen agglutination kits, PathoDx (Remel) (93%), Prolex (Pro-Lab Diagnostics) (38%), and Streptex (Remel) (53%). New approaches to bacterial identification in routine clinical microbiology laboratories may affect the prevalence of S. pseudoporcinus. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  14. Identification by Molecular Methods and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry and Antifungal Susceptibility Profiles of Clinically Significant Rare Aspergillus Species in a Referral Chest Hospital in Delhi, India.

    Masih, Aradhana; Singh, Pradeep K; Kathuria, Shallu; Agarwal, Kshitij; Meis, Jacques F; Chowdhary, Anuradha

    2016-09-01

    Aspergillus species cause a wide spectrum of clinical infections. Although Aspergillus fumigatus and Aspergillus flavus remain the most commonly isolated species in aspergillosis, in the last decade, rare and cryptic Aspergillus species have emerged in diverse clinical settings. The present study analyzed the distribution and in vitro antifungal susceptibility profiles of rare Aspergillus species in clinical samples from patients with suspected aspergillosis in 8 medical centers in India. Further, a matrix-assisted laser desorption ionization-time of flight mass spectrometry in-house database was developed to identify these clinically relevant Aspergillus species. β-Tubulin and calmodulin gene sequencing identified 45 rare Aspergillus isolates to the species level, except for a solitary isolate. They included 23 less common Aspergillus species belonging to 12 sections, mainly in Circumdati, Nidulantes, Flavi, Terrei, Versicolores, Aspergillus, and Nigri Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identified only 8 (38%) of the 23 rare Aspergillus isolates to the species level. Following the creation of an in-house database with the remaining 14 species not available in the Bruker database, the MALDI-TOF MS identification rate increased to 95%. Overall, high MICs of ≥2 μg/ml were noted for amphotericin B in 29% of the rare Aspergillus species, followed by voriconazole in 20% and isavuconazole in 7%, whereas MICs of >0.5 μg/ml for posaconazole were observed in 15% of the isolates. Regarding the clinical diagnoses in 45 patients with positive rare Aspergillus species cultures, 19 (42%) were regarded to represent colonization. In the remaining 26 patients, rare Aspergillus species were the etiologic agent of invasive, chronic, and allergic bronchopulmonary aspergillosis, allergic fungal rhinosinusitis, keratitis, and mycetoma. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  15. Identification by Molecular Methods and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and Antifungal Susceptibility Profiles of Clinically Significant Rare Aspergillus Species in a Referral Chest Hospital in Delhi, India

    Masih, Aradhana; Singh, Pradeep K.; Kathuria, Shallu; Agarwal, Kshitij

    2016-01-01

    Aspergillus species cause a wide spectrum of clinical infections. Although Aspergillus fumigatus and Aspergillus flavus remain the most commonly isolated species in aspergillosis, in the last decade, rare and cryptic Aspergillus species have emerged in diverse clinical settings. The present study analyzed the distribution and in vitro antifungal susceptibility profiles of rare Aspergillus species in clinical samples from patients with suspected aspergillosis in 8 medical centers in India. Further, a matrix-assisted laser desorption ionization–time of flight mass spectrometry in-house database was developed to identify these clinically relevant Aspergillus species. β-Tubulin and calmodulin gene sequencing identified 45 rare Aspergillus isolates to the species level, except for a solitary isolate. They included 23 less common Aspergillus species belonging to 12 sections, mainly in Circumdati, Nidulantes, Flavi, Terrei, Versicolores, Aspergillus, and Nigri. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) identified only 8 (38%) of the 23 rare Aspergillus isolates to the species level. Following the creation of an in-house database with the remaining 14 species not available in the Bruker database, the MALDI-TOF MS identification rate increased to 95%. Overall, high MICs of ≥2 μg/ml were noted for amphotericin B in 29% of the rare Aspergillus species, followed by voriconazole in 20% and isavuconazole in 7%, whereas MICs of >0.5 μg/ml for posaconazole were observed in 15% of the isolates. Regarding the clinical diagnoses in 45 patients with positive rare Aspergillus species cultures, 19 (42%) were regarded to represent colonization. In the remaining 26 patients, rare Aspergillus species were the etiologic agent of invasive, chronic, and allergic bronchopulmonary aspergillosis, allergic fungal rhinosinusitis, keratitis, and mycetoma. PMID:27413188

  16. Matrix-assisted laser desorption/ionization coupled with quadrupole/orthogonal acceleration time-of-flight mass spectrometry for protein discovery, identification, and structural analysis.

    Baldwin, M A; Medzihradszky, K F; Lock, C M; Fisher, B; Settineri, T A; Burlingame, A L

    2001-04-15

    The design and operation of a novel UV-MALDI ionization source on a commercial QqoaTOF mass spectrometer (Applied Biosystem/MDS Sciex QSTAR Pulsar) is described. Samples are loaded on a 96-well target plate, the movement of which is under software control and can be readily automated. Unlike conventional high-energy MALDI-TOF, the ions are produced with low energies (5-10 eV) in a region of relatively low vacuum (8 mTorr). Thus, they are cooled by extensive low-energy collisions before selection in the quadrupole mass analyzer (Q1), potentially giving a quasi-continuous ion beam ideally suited to the oaTOF used for mass analysis of the fragment ions, although ion yields from individual laser shots may vary widely. Ion dissociation is induced by collisions with argon in an rf-only quadrupole cell, giving typical low-energy CID spectra for protonated peptide ions. Ions separated in the oaTOF are registered by a four-anode detector and time-to-digital converter and accumulated in "bins" that are 625 ps wide. Peak shapes depend upon the number of ion counts in adjacent bins. As expected, the accuracy of mass measurement is shown to be dependent upon the number of ions recorded for a particular peak. With internal calibration, mass accuracy better than 10 ppm is attainable for peaks that contain sufficient ions to give well-defined Gaussian profiles. By virtue of its high resolution, capability for accurate mass measurements, and sensitivity in the low-femotomole range, this instrument is ideally suited to protein identification for proteomic applications by generation of peptide tags, manual sequence interpretation, identification of modifications such as phosphorylation, and protein structural elucidation. Unlike the multiply charged ions typical of electrospray ionization, the singly charged MALDI-generated peptide ions show a linear dependence of optimal collision energy upon molecular mass, which is advantageous for automated operation. It is shown that the novel

  17. Laser beam diagnostics for kilowatt power pulsed YAG laser

    Liu, Yi; Leong, Keng H.

    1992-01-01

    There is a growing need for high power YAG laser beam diagnostics with the recent introduction of such lasers in laser material processing. In this paper, we will describe the use of a commercially available laser beam analyzer (Prometec) to profile the laser beam from a 1600 W pulsed Nd:YAG laser that has a 1 mm fiber optic beam delivery system. The selection of laser pulse frequency and pulse width for the measurement is discussed. Laser beam propagation parameters by various optical components such as fibers and lenses can be determined from measurements using this device. The importance of such measurements will be discussed

  18. Laser ablation comparison by picosecond pulses train and nanosecond pulse

    Lednev, V. N.; Filippov, M. N.; Bunkin, A. F.; Pershin, S. M.

    2015-12-01

    A comparison of laser ablation by a train of picosecond pulses and nanosecond pulses revealed a difference in laser craters, ablation thresholds, plasma sizes and spectral line intensities. Laser ablation with a train of picosecond pulses resulted in improved crater quality while ablated mass decreased up to 30%. A reduction in laser plasma dimensions for picosecond train ablation was observed while the intensity of atomic/ionic lines in the plasma spectra was greater by a factor of 2-4 indicating an improved excitation and atomization in the plasma.

  19. System for increasing laser pulse rate

    1980-01-01

    A technique of static elements is disclosed for combining a plurality of laser beams having time sequenced, pulsed radiation to achieve an augmented pulse rate. The technique may also be applied in a system for combining both time sequenced pulses and frequency distinct pulses for use in a system for isotope enrichment. (author)

  20. Photoemission using femtosecond laser pulses

    Srinivasan-Rao, T.; Tsang, T.; Fischer, J.

    1991-10-01

    Successful operation of short wavelength FEL requires an electron bunch of current >100 A and normalized emittance < 1 mm-mrad. Recent experiments show that RF guns with photocathodes as the electron source may be the ideal candidate for achieving these parameters. To reduce the emittance growth due to space charge and RF dynamics effects, the gun may have to operate at high field gradient (hence at high RF frequency) and a spot size small compared to the aperture. This may necessitate the laser pulse duration to be in the subpicosecond regime to reduce the energy spread. We will present the behavior of metal photocathodes upon irradiation with femtosecond laser beams, comparison of linear and nonlinear photoemission, and scalability to high currents. Theoretical estimate of the intrinsic emittance at the photocathode in the presence of the anomalous heating of the electrons, and the tolerance on the surface roughness of the cathode material will be discussed

  1. Antimicrobial activity of biopolymer–antibiotic thin films fabricated by advanced pulsed laser methods

    Cristescu, R., E-mail: rodica.cristescu@inflpr.ro [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Popescu, C.; Dorcioman, G.; Miroiu, F.M.; Socol, G.; Mihailescu, I.N. [National Institute for Lasers, Plasma and Radiation Physics, Lasers Department, P.O. Box MG-36, Bucharest-Magurele (Romania); Gittard, S.D.; Miller, P.R.; Narayan, R.J. [Biomedical Engineering, University of North Carolina and North Carolina State University, Chapel Hill, NC 27599-7575 (United States); Enculescu, M. [National Institute for Materials Physics, PO Box MG-7, Bucharest-Magurele (Romania); Chrisey, D.B. [Tulane University, Department of Physics and Engineering Physics, New Orleans, LA (United States)

    2013-08-01

    We report on thin film deposition by matrix assisted pulsed laser evaporation (MAPLE) of two polymer–drug composite thin film systems. A pulsed KrF* excimer laser source (λ = 248 nm, τ = 25 ns, ν = 10 Hz) was used to deposit composite thin films of poly(D,L-lactide) (PDLLA) containing several gentamicin concentrations. FTIR spectroscopy was used to demonstrate that MAPLE-transferred materials exhibited chemical structures similar to those of drop cast materials. Scanning electron microscopy data indicated that MAPLE may be used to fabricate thin films of good morphological quality. The activity of PDLLA–gentamicin composite thin films against Staphylococcus aureus bacteria was demonstrated using drop testing. The influence of drug concentration on microbial viability was also assessed. Our studies indicate that polymer–drug composite thin films prepared by MAPLE may be used to impart antimicrobial activity to implants, medical devices, and other contact surfaces.

  2. Reactive pulsed laser deposition with gas jet

    Rakowski, R.; Bartnik, A.; Fiedorowicz, H.; Jarocki, R.; Kostecki, J.; Szczurek, M.

    2001-01-01

    Different metal (Sn, Al, steel, Cu, W) thin films were synthesized by reactive pulsed laser deposition on steel, copper and glass wafers. In our work pulsed Nd:glass (10 J, 800μs) laser system was used. Jet of gas was created by electromagnetic valve perpendicularly to the laser beam. Nitrogen, oxygen and argon were used. We used several to tens laser shots to obtain visible with the naked eye layers. Thin layers were observed under an optical microscope. (author)

  3. Evaluation of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry in comparison to rpoB gene sequencing for species identification of bloodstream infection staphylococcal isolates.

    Spanu, T; De Carolis, E; Fiori, B; Sanguinetti, M; D'Inzeo, T; Fadda, G; Posteraro, B

    2011-01-01

    As a result of variable expression of biochemical characters, misidentification by conventional phenotypic means often occurs with clinical isolates belonging to Staphylococcus species. Therefore, we evaluated the use of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of 450 blood isolates of the most relevant staphylococcal species, using sequence analysis of the rpoB gene as the reference method. A correct species identification by MALDI-TOF was obtained in 99.3% (447/450), with only three isolates being misidentified. In addition, MALDI-TOF correctly identified all the staphylococcal subspecies studied, including Staphylococcus capitis subsp. capitis and subsp. urealyticus, Staphylococcus cohnii subsp. urealyticus, Staphylococcus hominis subsp. novobiosepticus and subsp. hominis, Staphylococcus saprophyticus subsp. saprophyticus, Staphylococcus schleiferi subsp. schleiferi and Staphylococcus sciuri subsp. sciuri. Thus, MALDI-TOF MS-based species identification of staphylococci can be routinely achieved without any substantial costs for consumables or the time needed for labour-intensive DNA sequence analysis. © 2010 The Authors. Journal Compilation © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  4. Heterotrophic monitoring at a drinking water treatment plant by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry after different drinking water treatments.

    Sala-Comorera, Laura; Blanch, Anicet R; Vilaró, Carles; Galofré, Belén; García-Aljaro, Cristina

    2017-10-01

    The aim of this work was to assess the suitability of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) for routine heterotrophic monitoring in a drinking water treatment plant. Water samples were collected from raw surface water and after different treatments during two campaigns over a 1-year period. Heterotrophic bacteria were studied and isolates were identified by MALDI-TOF MS. Moreover, the diversity index and the coefficient of population similarity were also calculated using biochemical fingerprinting of the populations studied. MALDI-TOF MS enabled us to characterize and detect changes in the bacterial community composition throughout the water treatment plant. Raw water showed a large and diverse population which was slightly modified after initial treatment steps (sand filtration and ultrafiltration). Reverse osmosis had a significant impact on the microbial diversity, while the final chlorination step produced a shift in the composition of the bacterial community. Although MALDI-TOF MS could not identify all the isolates since the available MALDI-TOF MS database does not cover all the bacterial diversity in water, this technique could be used to monitor bacterial changes in drinking water treatment plants by creating a specific protein profile database for tracking purposes.

  5. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

    Lamer, S; Jungblut, P R

    2001-03-10

    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

  6. Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing.

    Gitman, Melissa R; McTaggart, Lisa; Spinato, Joanna; Poopalarajah, Rahgavi; Lister, Erin; Husain, Shahid; Kus, Julianne V

    2017-07-01

    Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods. © Crown copyright 2017.

  7. Evaluation of the Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Aspergillus Species Directly from Growth on Solid Agar Media

    Ying Li

    2017-06-01

    Full Text Available We evaluated the accuracy of the Bruker Biotyper matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS system at identifying clinical isolates of Aspergillus species that were grown on agar media. A total of 381 non-duplicate Aspergillus isolates representing 21 different Aspergillus species identified by molecular analysis were included in this study. The Bruker Biotyper MALDI-TOF MS system was able to identify 30.2% (115/381 of the isolates to the species level (score values of ≥2.000 and 49.3% to the genus level (score values of 1.700–1.999. When the identification cutoff value was lowered from ≥2.000 to ≥1.700, the species-level identification rate increased to 79.5% with a slight rise of false identification from 2.6 to 5.0%. From another aspect, a correct species-level identification rate of 89% could be reached by the Bruker Biotyper MALDI-TOF MS system regardless of the score values obtained. The Bruker Biotyper MALDI-TOF MS system had a moderate performance in identification of Aspergillus directly inoculated on solid agar media. Continued expansion of the Bruker Biotyper MALDI-TOF MS database and adoption of alternative cutoff values for interpretation are required to improve the performance of the system for identifying highly diverse species of clinically encountered Aspergillus isolates.

  8. Evaluation of the Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Aspergillus Species Directly from Growth on Solid Agar Media.

    Li, Ying; Wang, He; Zhao, Yu-Pei; Xu, Ying-Chun; Hsueh, Po-Ren

    2017-01-01

    We evaluated the accuracy of the Bruker Biotyper matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) system at identifying clinical isolates of Aspergillus species that were grown on agar media. A total of 381 non-duplicate Aspergillus isolates representing 21 different Aspergillus species identified by molecular analysis were included in this study. The Bruker Biotyper MALDI-TOF MS system was able to identify 30.2% (115/381) of the isolates to the species level (score values of ≥2.000) and 49.3% to the genus level (score values of 1.700-1.999). When the identification cutoff value was lowered from ≥2.000 to ≥1.700, the species-level identification rate increased to 79.5% with a slight rise of false identification from 2.6 to 5.0%. From another aspect, a correct species-level identification rate of 89% could be reached by the Bruker Biotyper MALDI-TOF MS system regardless of the score values obtained. The Bruker Biotyper MALDI-TOF MS system had a moderate performance in identification of Aspergillus directly inoculated on solid agar media. Continued expansion of the Bruker Biotyper MALDI-TOF MS database and adoption of alternative cutoff values for interpretation are required to improve the performance of the system for identifying highly diverse species of clinically encountered Aspergillus isolates.

  9. Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing

    Gitman, Melissa R.; McTaggart, Lisa; Spinato, Joanna; Poopalarajah, Rahgavi; Lister, Erin; Husain, Shahid

    2017-01-01

    ABSTRACT Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods. PMID:28404678

  10. Imipenem-avibactam: a novel combination for the rapid detection of carbapenemase activity in Enterobacteriaceae and Acinetobacter baumannii by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Oviaño, Marina; Bou, Germán

    2017-02-01

    In the present study, we propose a novel matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for detecting carbapenemase-producing Enterobacteriaceae and Acinetobacter baumannii. For this, we analyzed a series of 131 isolates. Among them, a total of 115 Enterobacteriaceae: 79 of them carrying a carbapenemase enzyme (15bla KPC , 7bla NDM , 11bla IMP , 12bla VIM , and 34bla OXA-48 ) and 16 A. baumannii isolates: 15 of them carrying carbapenemases (10bla OXA-23, 2bla OXA-58, 2bla OXA-24 , and 1bla OXA-237 ). The rest of the isolates were noncarbapenemase producers and used as negative controls. The isolates were submitted to susceptibility testing using a combination of imipenem-avibactam and analysis by the MALDI-TOF Biotyper Compass software (Bruker Daltonik, Germany). The assay showed an overall sensitivity and specificity for carbapenemase detection of 98% and 100%, respectively. The combination of imipenem and avibactam displayed activity against KPC and OXA-48-producing Enterobacteriaceae and thus represents a new strategy for identifying and confirming these carbapenemases. However, the combination did not provide any benefit over A. baumannii. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Methylobacterium Species Promoting Rice and Barley Growth and Interaction Specificity Revealed with Whole-Cell Matrix-Assisted Laser Desorption/Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF/MS Analysis.

    Akio Tani

    Full Text Available Methylobacterium species frequently inhabit plant surfaces and are able to utilize the methanol emitted from plants as carbon and energy sources. As some of the Methylobacterium species are known to promote plant growth, significant attention has been paid to the mechanism of growth promotion and the specificity of plant-microbe interactions. By screening our Methylobacterium isolate collection for the high growth promotion effect in vitro, we selected some candidates for field and pot growth tests for rice and barley, respectively. We found that inoculation resulted in better ripening of rice seeds, and increased the size of barley grains but not the total yield. In addition, using whole-cell matrix-assister laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS analysis, we identified and classified Methylobacterium isolates from Methylobacterium-inoculated rice plants. The inoculated species could not be recovered from the rice plants, and in some cases, the Methylobacterium community structure was affected by the inoculation, but not with predomination of the inoculated species. The isolates from non-inoculated barley of various cultivars grown in the same field fell into just two species. These results suggest that there is a strong selection pressure at the species level of Methylobacterium residing on a given plant species, and that selection of appropriate species that can persist on the plant is important to achieve growth promotion.

  12. Comparative evaluation of matrix-assisted laser desorption ionisation-time of flight mass spectrometry and conventional phenotypic-based methods for identification of clinically important yeasts in a UK-based medical microbiology laboratory.

    Fatania, Nita; Fraser, Mark; Savage, Mike; Hart, Jason; Abdolrasouli, Alireza

    2015-12-01

    Performance of matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) was compared in a side-by side-analysis with conventional phenotypic methods currently in use in our laboratory for identification of yeasts in a routine diagnostic setting. A diverse collection of 200 clinically important yeasts (19 species, five genera) were identified by both methods using standard protocols. Discordant or unreliable identifications were resolved by sequencing of the internal transcribed spacer region of the rRNA gene. MALDI-TOF and conventional methods were in agreement for 182 isolates (91%) with correct identification to species level. Eighteen discordant results (9%) were due to rarely encountered species, hence the difficulty in their identification using traditional phenotypic methods. MALDI-TOF MS enabled rapid, reliable and accurate identification of clinically important yeasts in a routine diagnostic microbiology laboratory. Isolates with rare, unusual or low probability identifications should be confirmed using robust molecular methods. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  13. Dansyl-peptides matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) and tandem mass spectrometric (MS/MS) features improve the liquid chromatography/MALDI-MS/MS analysis of the proteome.

    Chiappetta, Giovanni; Ndiaye, Sega; Demey, Emmanuelle; Haddad, Iman; Marino, Gennaro; Amoresano, Angela; Vinh, Joëlle

    2010-10-30

    Peptide tagging is a useful tool to improve matrix-assisted laser desorption/ionization tandem mass spectrometric (MALDI-MS/MS) analysis. We present a new application of the use of the dansyl chloride (DNS-Cl). DNS-Cl is a specific primary amine reagent widely used in protein biochemistry. It adds a fluorescent dimethylaminonaphthalene moiety to the molecule. The evaluation of MALDI-MS and MS/MS analyses of dansylated peptides shows that dansylation raises the ionization efficiency of the most hydrophilic species compared with the most hydrophobic ones. Consequently, higher Mascot scores and protein sequence coverage are obtained by combining MS and MS/MS data of native and tagged samples. The N-terminal DNS-Cl sulfonation improves the peptide fragmentation and promotes the generation of b-fragments allowing better peptide sequencing. In addition, we set up a labeling protocol based on the microwave chemistry. Peptide dansylation proved to be a rapid and cheap method to improve the performance of liquid chromatography (LC)/MALDI-MS/MS analysis at the proteomic scale in terms of peptide detection and sequence coverage. Copyright © 2010 John Wiley & Sons, Ltd.

  14. Metabolite localization by atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging in whole-body sections and individual organs of the rove beetle Paederus riparius.

    Bhandari, Dhaka Ram; Schott, Matthias; Römpp, Andreas; Vilcinskas, Andreas; Spengler, Bernhard

    2015-03-01

    Mass spectrometry imaging provides for non-targeted, label-free chemical imaging. In this study, atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) was used for the first time to describe the chemical distribution of the defensive compounds pederin, pseudopederin, and pederon in tissue sections (16 μm thick) of the rove beetle Paederus riparius. The whole-insect tissue section was scanned with a 20-μm step size. Mass resolution of the orbital trapping mass spectrometer was set to 100,000 at m/z 200. Additionally, organ-specific compounds were identified for brain, nerve cord, eggs, gut, ovaries, and malpighian tubules. To confirm the distribution of the specific compounds, individual organs from the insect were dissected, and MSI experiments were performed on the dissected organs. Three ganglia of the nerve cord, with a dimension of 250-500 μm, were measured with 10-μm spatial resolution. High-quality m/z images, based on high spatial resolution and high mass accuracy were generated. These features helped to assign mass spectral peaks with high confidence. Mass accuracy of the imaging experiments was section. Without any labeling, we assigned key lipids for specific organs to describe their location in the body and to identify morphological structures with a specificity higher than with staining or immunohistology methods.

  15. Identification and Quantification of N-Acyl Homoserine Lactones Involved in Bacterial Communication by Small-Scale Synthesis of Internal Standards and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

    Leipert, Jan; Treitz, Christian; Leippe, Matthias; Tholey, Andreas

    2017-12-01

    N-acyl homoserine lactones (AHL) are small signal molecules involved in the quorum sensing of many gram-negative bacteria, and play an important role in biofilm formation and pathogenesis. Present analytical methods for identification and quantification of AHL require time-consuming sample preparation steps and are hampered by the lack of appropriate standards. By aiming at a fast and straightforward method for AHL analytics, we investigated the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Suitable MALDI matrices, including crystalline and ionic liquid matrices, were tested and the fragmentation of different AHL in collision-induced dissociation MS/MS was studied, providing information about characteristic marker fragments ions. Employing small-scale synthesis protocols, we established a versatile and cost-efficient procedure for fast generation of isotope-labeled AHL standards, which can be used without extensive purification and yielded accurate standard curves. Quantitative analysis was possible in the low pico-molar range, with lower limits of quantification reaching from 1 to 5 pmol for different AHL. The developed methodology was successfully applied in a quantitative MALDI MS analysis of low-volume culture supernatants of Pseudomonas aeruginosa. [Figure not available: see fulltext.

  16. A new scheme for strain typing of methicillin-resistant Staphylococcus aureus on the basis of matrix-assisted laser desorption ionization time-of-flight mass spectrometry by using machine learning approach.

    Wang, Hsin-Yao; Lee, Tzong-Yi; Tseng, Yi-Ju; Liu, Tsui-Ping; Huang, Kai-Yao; Chang, Yung-Ta; Chen, Chun-Hsien; Lu, Jang-Jih

    2018-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA), one of the most important clinical pathogens, conducts an increasing number of morbidity and mortality in the world. Rapid and accurate strain typing of bacteria would facilitate epidemiological investigation and infection control in near real time. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry is a rapid and cost-effective tool for presumptive strain typing. To develop robust method for strain typing based on MALDI-TOF spectrum, machine learning (ML) is a promising algorithm for the construction of predictive model. In this study, a strategy of building templates of specific types was used to facilitate generating predictive models of methicillin-resistant Staphylococcus aureus (MRSA) strain typing through various ML methods. The strain types of the isolates were determined through multilocus sequence typing (MLST). The area under the receiver operating characteristic curve (AUC) and the predictive accuracy of the models were compared. ST5, ST59, and ST239 were the major MLST types, and ST45 was the minor type. For binary classification, the AUC values of various ML methods ranged from 0.76 to 0.99 for ST5, ST59, and ST239 types. In multiclass classification, the predictive accuracy of all generated models was more than 0.83. This study has demonstrated that ML methods can serve as a cost-effective and promising tool that provides preliminary strain typing information about major MRSA lineages on the basis of MALDI-TOF spectra.

  17. Fingerprinting of egg and oil binders in painted artworks by matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of lipid oxidation by-products.

    Calvano, C D; van der Werf, I D; Palmisano, F; Sabbatini, L

    2011-06-01

    A matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based approach was applied for the detection of various lipid classes, such as triacylglycerols (TAGs) and phospholipids (PLs), and their oxidation by-products in extracts of small (50-100 μg) samples obtained from painted artworks. Ageing of test specimens under various conditions, including the presence of different pigments, was preliminarily investigated. During ageing, the TAGs and PLs content decreased, whereas the amount of diglycerides, short-chain oxidative products arising from TAGs and PLs, and oxidized TAGs and PLs components increased. The examination of a series of model paint samples gave a clear indication that specific ions produced by oxidative cleavage of PLs and/or TAGs may be used as markers for egg and drying oil-based binders. Their elemental composition and hypothetical structure are also tentatively proposed. Moreover, the simultaneous presence of egg and oil binders can be easily and unambiguously ascertained through the simultaneous occurrence of the relevant specific markers. The potential of the proposed approach was demonstrated for the first time by the analysis of real samples from a polyptych of Bartolomeo Vivarini (fifteenth century) and a "French school" canvas painting (seventeenth century).

  18. Evaluation of repetitive-PCR and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS for rapid strain typing of Bacillus coagulans.

    Jun Sato

    Full Text Available In order to establish rapid and accurate typing method for Bacillus coagulans strains which is important for controlling in some canned foods and tea-based beverages manufacturing because of the high-heat resistance of the spores and high tolerance of the vegetative cells to catechins and chemicals, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS and repetitive-PCR (rep-PCR were evaluated. For this purpose, 28 strains of B. coagulans obtained from various culture collections were tested. DNA sequence analyses of the genes encoding 16S rRNA and DNA gyrase classified the test strains into two and three groups, respectively, regardless of their phenotypes. Both MALDI-TOF MS and rep-PCR methods classified the test strains in great detail. Strains classified in each group showed similar phenotypes, such as carbohydrate utilization determined using API 50CH. In particular, the respective two pairs of strains which showed the same metabolic characteristic were classified into the same group by both MALDI-TOF MS and rep-PCR methods separating from the other strains. On the other hand, the other strains which have the different profiles of carbohydrate utilization were separated into different groups by these methods. These results suggested that the combination of MALDI-TOF MS and rep-PCR analyses was advantageous for the rapid and detailed typing of bacterial strains in respect to both phenotype and genotype.

  19. Evaluation of repetitive-PCR and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for rapid strain typing of Bacillus coagulans.

    Sato, Jun; Nakayama, Motokazu; Tomita, Ayumi; Sonoda, Takumi; Hasumi, Motomitsu; Miyamoto, Takahisa

    2017-01-01

    In order to establish rapid and accurate typing method for Bacillus coagulans strains which is important for controlling in some canned foods and tea-based beverages manufacturing because of the high-heat resistance of the spores and high tolerance of the vegetative cells to catechins and chemicals, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and repetitive-PCR (rep-PCR) were evaluated. For this purpose, 28 strains of B. coagulans obtained from various culture collections were tested. DNA sequence analyses of the genes encoding 16S rRNA and DNA gyrase classified the test strains into two and three groups, respectively, regardless of their phenotypes. Both MALDI-TOF MS and rep-PCR methods classified the test strains in great detail. Strains classified in each group showed similar phenotypes, such as carbohydrate utilization determined using API 50CH. In particular, the respective two pairs of strains which showed the same metabolic characteristic were classified into the same group by both MALDI-TOF MS and rep-PCR methods separating from the other strains. On the other hand, the other strains which have the different profiles of carbohydrate utilization were separated into different groups by these methods. These results suggested that the combination of MALDI-TOF MS and rep-PCR analyses was advantageous for the rapid and detailed typing of bacterial strains in respect to both phenotype and genotype.

  20. Microorganisms in cryopreserved semen and culture media used in the in vitro production (IVP) of bovine embryos identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS).

    Zampieri, Dávila; Santos, Vanessa G; Braga, Patrícia A C; Ferreira, Christina R; Ballottin, Daniela; Tasic, Ljubica; Basso, Andréa C; Sanches, Bruno V; Pontes, José H F; da Silva, Bárbara Pereira; Garboggini, Fabiana Fantinatti; Eberlin, Marcos N; Tata, Alessandra

    2013-09-01

    Commercial cattle breeders produce their own herd offspring for the dairy and beef market using artificial insemination. The procedure involves sanitary risks associated with the collection and commercialization of the germplasm, and the in vitro production and transfer of the bovine embryos must be monitored by strict health surveillance. To avoid the spreading of infectious diseases, one must rely on using controlled and monitored germplasm, media, and reagents that are guaranteed free of pathogens. In this article, we investigated the use of a new mass spectrometric approach for fast and accurate identification of bacteria and fungi in bovine semen and in culture media employed in the embryo in vitro production process. The microorganisms isolated from samples obtained in a commercial bovine embryo IVP setting were identified in a few minutes by their conserved peptide/protein profile, obtained applying matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), matched against a commercial database. The successful microorganisms MS identification has been confirmed by DNA amplification and sequencing. Therefore, the MS technique seems to offer a powerful tool for rapid and accurate microorganism identification in semen and culture media samples. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. Capitalizing Resolving Power of Density Gradient Ultracentrifugation by Freezing and Precisely Slicing Centrifuged Solution: Enabling Identification of Complex Proteins from Mitochondria by Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

    Haiqing Yu

    2016-01-01

    Full Text Available Density gradient centrifugation is widely utilized for various high purity sample preparations, and density gradient ultracentrifugation (DGU is often used for more resolution-demanding purification of organelles and protein complexes. Accurately locating different isopycnic layers and precisely extracting solutions from these layers play a critical role in achieving high-resolution DGU separations. In this technique note, we develop a DGU procedure by freezing the solution rapidly (but gently after centrifugation to fix the resolved layers and by slicing the frozen solution to fractionate the sample. Because the thickness of each slice can be controlled to be as thin as 10 micrometers, we retain virtually all the resolution produced by DGU. To demonstrate the effectiveness of this method, we fractionate complex V from HeLa mitochondria using a conventional technique and this freezing-slicing (F-S method. The comparison indicates that our F-S method can reduce complex V layer thicknesses by ~40%. After fractionation, we analyze complex V proteins directly on a matrix assisted laser desorption/ionization, time-of-flight mass spectrometer. Twelve out of fifteen subunits of complex V are positively identified. Our method provides a practical protocol to identify proteins from complexes, which is useful to investigate biomolecular complexes and pathways in various conditions and cell types.

  2. Matrix-assisted ultraviolet laser desorption/ionization time-of-flight mass spectrometry of beta-(1 --> 3), beta-(1 --> 4)-xylans from Nothogenia fastigiata using nor-harmane as matrix.

    Fukuyama, Yuko; Kolender, Adriana A; Nishioka, Masae; Nonami, Hiroshi; Matulewicz, María C; Erra-Balsells, Rosa; Cerezo, Alberto S

    2005-01-01

    Three xylan fractions isolated from the red seaweed Nothogenia fastigiata (Nemaliales) were analyzed by ultraviolet matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (UV-MALDI-TOFMS). UV-MALDI-TOFMS was carried out in the linear and reflectron modes, and as routine in the positive and negative ion modes. Of the several matrices tested, nor-harmane was the only effective one giving good spectra in the positive ion mode. The number-average molar masses of two of the fractions, calculated from the distribution profiles, were lower than those determined previously by (1)H NMR analysis, suggesting a decrease in the ionization efficiency with increasing molecular weight; weight-average molar mass and polydispersity index were also determined. As the xylans retained small but significant quantities of calcium salts, the influence of added Ca(2+) as CaCl(2) on UV-MALDI-MS was investigated. The simultaneous addition of sodium chloride and calcium chloride was also analyzed. Addition of sodium chloride did not change the distribution profile of the native sample showing that the inhibitory effect is due to Ca(2+) and not to Cl(-). Addition of calcium chloride with 1:1 analyte/salt molar ratio gave spectra with less efficient desorption/ionization of oligomers; the signals of these oligomers were completely suppressed when the addition of the salt became massive (1:100 analyte/salt molar ratio). Copyright (c) 2005 John Wiley & Sons, Ltd.

  3. New strategy for the determination of gliadins in maize- or rice-based foods matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: fractionation of gliadins from maize or rice prolamins by acidic treatment.

    Hernando, Alberto; Valdes, Israel; Méndez, Enrique

    2003-08-01

    A procedure for determining small quantities of gliadins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) in gluten-free foods containing relatively large amounts of prolamin proteins from maize or rice is described. We report for the first time that gliadins, the ethanol-soluble wheat prolamin fraction, can be quantitatively solubilized in 1.0 M acetic acid, while the corresponding ethanol-soluble maize or rice prolamin fraction remains insoluble in acetic acid. We describe a methodology for the detection of gliadins in maize and rice foods based on a two-step procedure of extraction (60% aqueous ethanol followed by 1 M acetic acid). Subsequent MALDI-TOFMS analysis of the resulting acidic extract from these gluten-free foods clearly confirms the presence of a typical mass pattern corresponding to gliadin components, ranging from 30 to 45 kDa. Depending on the percentages of maize or rice flours employed in the elaboration of these foods, the combined procedure enables levels of gliadins from 100 to 400 ppm to be detected. The efficiency of this combined procedure corroborates enzyme-linked immunosorbent assay data for a large number of maize/rice gluten-free foods by means of direct visualization of the characteristic gliadin mass pattern in maize or rice foods. Copyright 2003 John Wiley & Sons, Ltd.

  4. Direct identification of microorganisms from positive blood cultures using the lysis-filtration technique and matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): a multicentre study.

    Farina, Claudio; Arena, Fabio; Casprini, Patrizia; Cichero, Paola; Clementi, Massimo; Cosentino, Marina; Degl'Innocenti, Roberto; Giani, Tommaso; Luzzaro, Francesco; Mattei, Romano; Mauri, Carola; Nardone, Maria; Rossolini, Gian Maria; Serna Ortega, Paula Andrea; Vailati, Francesca

    2015-04-01

    Microbial identification from blood cultures is essential to institute optimal antibiotic therapy and improve survival possibilities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacteria and yeasts from positive blood cultures broths. The aim of this multicentre study was to evaluate the reliability of the lysis-filtration technique associated with MALDI-TOF MS to directly identify microorganisms from 765 positive blood cultures collected in six Italian hospitals. Overall, 675/765 (78.1%) blood isolates were correctly identified at the species level, with significant differences between Gram-negative and Gram-positive bacteria (92.6%, and 69.8%, respectively). Some difficulties arise in identifying Streptococcus pneumoniae, Staphylococcus aureus, yeasts and anaerobes. The lysis-filtration protocol is a suitable procedure in terms of performance in identifying microorganisms, but it is quite expensive and technically time-consuming since the time of filtration is not regular for all the samples. The application of the MALDI-TOF MS technique to the direct microbial identification from positive blood cultures is a very promising approach, even if more experience must be gained to minimize errors and costs.

  5. Novel, improved sample preparation for rapid, direct identification from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.

    Schubert, Sören; Weinert, Kirsten; Wagner, Chris; Gunzl, Beatrix; Wieser, Andreas; Maier, Thomas; Kostrzewa, Markus

    2011-11-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used for rapid and reliable identification of bacteria and yeast grown on agar plates. Moreover, MALDI-TOF MS also holds promise for bacterial identification from blood culture (BC) broths in hospital laboratories. The most important technical step for the identification of bacteria from positive BCs by MALDI-TOF MS is sample preparation to remove blood cells and host proteins. We present a method for novel, rapid sample preparation using differential lysis of blood cells. We demonstrate the efficacy and ease of use of this sample preparation and subsequent MALDI-TOF MS identification, applying it to a total of 500 aerobic and anaerobic BCs reported to be positive by a Bactec 9240 system. In 86.5% of all BCs, the microorganism species were correctly identified. Moreover, in 18/27 mixed cultures at least one isolate was correctly identified. A novel method that adjusts the score value for MALDI-TOF MS results is proposed, further improving the proportion of correctly identified samples. The results of the present study show that the MALDI-TOF MS-based method allows rapid (directly from positive BCs and with high accuracy. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  6. Direct identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS from positive blood culture bottles: An opportunity to customize growth conditions for fastidious organisms causing bloodstream infections

    Megha Sharma

    2017-01-01

    Full Text Available Culture-negative bacteraemia has been an enigmatic entity with respect to its aetiological agents. In an attempt to actively identify those positive blood cultures that escape isolation and detection on routine workflow, an additional step of MALDI-TOF MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry based detection was carried out directly from the flagged blood culture bottles. Blood samples from 200 blood culture bottles that beeped positive with automated (BACTEC system and showed no growth of organism on routine culture media, were subjected to analysis by MALDI-TOF MS. Forty seven of the 200 (23.5% bacterial aetiology could be established by bottle-based method. Based on these results, growth on culture medium could be achieved for the isolates by providing special growth conditions to the fastidious organisms. Direct identification by MALDI-TOF MS from BACTEC-positive bottles provided an opportunity to isolate those fastidious organisms that failed to grow on routine culture medium by providing them with necessary alterations in growth environment.

  7. Direct identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) from positive blood culture bottles: An opportunity to customize growth conditions for fastidious organisms causing bloodstream infections.

    Sharma, Megha; Gautam, Vikas; Mahajan, Monika; Rana, Sudesh; Majumdar, Manasi; Ray, Pallab

    2017-10-01

    Culture-negative bacteraemia has been an enigmatic entity with respect to its aetiological agents. In an attempt to actively identify those positive blood cultures that escape isolation and detection on routine workflow, an additional step of MALDI-TOF MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry) based detection was carried out directly from the flagged blood culture bottles. Blood samples from 200 blood culture bottles that beeped positive with automated (BACTEC) system and showed no growth of organism on routine culture media, were subjected to analysis by MALDI-TOF MS. Forty seven of the 200 (23.5%) bacterial aetiology could be established by bottle-based method. Based on these results, growth on culture medium could be achieved for the isolates by providing special growth conditions to the fastidious organisms. Direct identification by MALDI-TOF MS from BACTEC-positive bottles provided an opportunity to isolate those fastidious organisms that failed to grow on routine culture medium by providing them with necessary alterations in growth environment.

  8. Development of an improved rapid BACpro® protocol and a method for direct identification from blood-culture-positive bottles using matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Yonezawa, Takatoshi; Watari, Tomohisa; Ashizawa, Kazuho; Hanada, Daisuke; Yanagiya, Takako; Watanabe, Naoki; Terada, Takashi; Tomoda, Yutaka; Fujii, Satoshi

    2018-05-01

    Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been incorporated into pathogenic bacterial identification methods and has improved their rapidity. Various methods have been reported to directly identify bacteria with MALDI-TOF MS by pretreating culture medium in blood culture bottles. Rapid BACpro® (Nittobo Medical Co., Ltd.) is a pretreatment kit for effective collection of bacteria with cationic copolymers. However, the Rapid BACpro® pretreatment kit is adapted only for MALDI Biotyper (Bruker Daltonics K.K.), and there has been a desire to expand its use to VITEK MS (VMS; bioMerieux SA). We improved the protocol and made it possible to analyze with VMS. The culture medium bacteria collection method was changed to a method with centrifugation after hemolysis using saponin; the cationic copolymer concentration was changed to 30% of the original concentration; the sequence with which reagents were added was changed; and a change was made to an ethanol/formic acid extraction method. The improved protocol enhanced the identification performance. When VMS was used, the identification rate was 100% with control samples. With clinical samples, the identification agreement rate with the cell smear method was 96.3%. The improved protocol is effective in blood culture rapid identification, being both simpler and having an improved identification performance compared with the original. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. Graphene/TiO2 nanocomposite based solid-phase extraction and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for lipidomic profiling of avocado (Persea americana Mill.).

    Shen, Qing; Yang, Mei; Li, Linqiu; Cheung, Hon-Yeung

    2014-12-10

    Phospholipids possess important physiological, structural and nutritional functions in biological systems. This study described a solid-phase extraction (SPE) method, employing graphene and titanium dioxide (G/TiO2) nanocomposite as sorbent, for the selective isolation and enrichment of phospholipids from avocado (Persea americana Mill.). Based on the principal that the phosphoryl group in the phospholipid can interact with TiO2 via a bridging bidentate mode, an optimum condition was established for SPE, and was successfully applied to prepare avocado samples. The extracts were monitored by matrix-assisted laser desorption ionization time-of-flight/tandem mass spectrometry (MALDI-TOF/MS) in both positive-ion and negative-ion modes. Results showed that phospholipids could be efficiently extracted in a clean manner by G/TiO2 based SPE. In addition, the signals of phospholipids were enhanced while the noise was reduced. Some minor peaks became more obvious. In conclusion, the nanocomposite material of G/TiO2 was proved to be a promising sorbent for selective separation of phospholipids from crude lipid extract. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  10. Genetic, phenotypic and matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based identification of anaerobic bacteria and determination of their antimicrobial susceptibility at a University Hospital in Japan.

    Yunoki, Tomoyuki; Matsumura, Yasufumi; Nakano, Satoshi; Kato, Karin; Hotta, Go; Noguchi, Taro; Yamamoto, Masaki; Nagao, Miki; Takakura, Shunji; Ichiyama, Satoshi

    2016-05-01

    The accuracies of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and the phenotypic method using VITEK 2 were compared to the accuracy of 16S rRNA sequence analysis for the identification of 170 clinically isolated anaerobes. The antimicrobial susceptibility of the isolates was also evaluated. Genetic analysis identified 21 Gram-positive species in 14 genera and 29 Gram-negative species in 11 genera. The most frequently isolated genera were Prevotella spp. (n = 46), Bacteroides spp. (n = 25) and Clostridium spp. (n = 25). MALDI-TOF MS correctly identified more isolates compared with VITEK 2 at the species (80 vs. 58%, respectively; p anaerobic agents indicated that the isolates of the three most frequently identified anaerobic genera exhibited good antimicrobial susceptibility. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  11. Analysis of picosecond pulsed laser melted graphite

    Steinbeck, J.; Braunstein, G.; Speck, J.; Dresselhaus, M.S.; Huang, C.Y.; Malvezzi, A.M.; Bloembergen, N.

    1986-01-01

    A Raman microprobe and high resolution TEM have been used to analyze the resolidified region of liquid carbon generated by picosecond pulse laser radiation. From the relative intensities of the zone center Raman-allowed mode for graphite at 1582 cm -1 and the disorder-induced mode at 1360 cm -1 , the average graphite crystallite size in the resolidified region is determined as a function of position. By comparison with Rutherford backscattering spectra and Raman spectra from nonosecond pulsed laser melting experiments, the disorder depth for picosecond pulsed laser melted graphite is determined as a function of irradiating energy density. Comparisons of TEM micrographs for nanosecond and picosecond pulsed laser melting experiments show that the structure of the laser disordered regions in graphite are similar and exhibit similar behavior with increasing laser pulse fluence

  12. Effect of laser pulse energies in laser induced breakdown spectroscopy in double-pulse configuration

    Benedetti, P.A.; Cristoforetti, G.; Legnaioli, S.; Palleschi, V.; Pardini, L.; Salvetti, A.; Tognoni, E.

    2005-01-01

    In this paper, the effect of laser pulse energy on double-pulse laser induced breakdown spectroscopy signal is studied. In particular, the energy of the first pulse has been changed, while the second pulse energy is held fixed. A systematic study of the laser induced breakdown spectroscopy signal dependence on the interpulse delay is performed, and the results are compared with the ones obtained with a single laser pulse of energy corresponding to the sum of the two pulses. At the same time, the crater formed at the target surface is studied by video-confocal microscopy, and the variation in crater dimensions is correlated to the enhancement of the laser induced breakdown spectroscopy signal. The results obtained are consistent with the interpretation of the double-pulse laser induced breakdown spectroscopy signal enhancement in terms of the changes in ambient gas pressure produced by the shock wave induced by the first laser pulse

  13. Using solvent-free sample preparation to promote protonation of poly(ethylene oxide)s with labile end-groups in matrix-assisted laser desorption/ionisation.

    Mazarin, Michael; Phan, Trang N T; Charles, Laurence

    2008-12-01

    Protonation is usually required to observe intact ions during matrix-assisted laser desorption/ionization (MALDI) of polymers containing fragile end-groups while cation adduction induces chain-end degradation. These polymers, generally obtained via living free radical polymerization techniques, are terminated with a functionality in which a bond is prone to homolytic cleavage, as required by the polymerization process. A solvent-free sample preparation method was used here to avoid salt contaminant from the solvent traditionally used in the dried-droplet MALDI procedure. Solvent-based and solvent-free sample preparations were compared for a series of three poly(ethylene oxide) polymers functionalized with a labile end-group in a nitroxide-mediated polymerization reaction, using 2,4,6-trihydroxyacetophenone (THAP) as the matrix without any added salt. Intact oligomer ions could only be produced as protonated molecules in solvent-free MALDI while sodium adducts of degraded polymers were formed from the dried-droplet samples. Although MALDI analysis was performed at the laser threshold, fragmentation of protonated macromolecules was still observed to occur. However, in contrast to sodiated molecules, dissociation of protonated oligomers does not involve the labile C--ON bond of the end-group. As the macromolecule size increased, protonation appeared to be less efficient and sodium adduction became the dominant ionization process, although no sodium salt was added in the preparation. Formation of sodiated degraded macromolecules would be dictated by increasing cation affinity as the size of the oligomers increases and would reveal the presence of salts at trace levels in the MALDI samples.

  14. Utility of imaging mass spectrometry (IMS) by matrix-assisted laser desorption ionization (MALDI) on an ion trap mass spectrometer in the analysis of drugs and metabolites in biological tissues.

    Drexler, Dieter M; Garrett, Timothy J; Cantone, Joseph L; Diters, Richard W; Mitroka, James G; Prieto Conaway, Maria C; Adams, Stephen P; Yost, Richard A; Sanders, Mark

    2007-01-01

    The properties and potential liabilities of drug candidate are investigated in detailed ADME assays and in toxicity studies, where findings are placed in context of exposure to dosed drug and metabolites. The complex nature of biological samples may necessitate work-up procedures prior to high performance liquid chromatography-mass spectrometric (HPLC-MS) analysis of endogenous or xenobiotic compounds. This concept can readily be applied to biological fluids such as blood or urine, but in localized samples such as organs and tissues potentially important spatial, thus anatomical, information is lost during sample preparation as the result of homogenization and extraction procedures. However, the localization of test article or spatial identification of metabolites may be critical to the understanding of the mechanism of target-organ toxicity and its relevance to clinical safety. Tissue imaging mass spectrometry (IMS) by matrix-assisted laser desorption ionization (MALDI) and ion trap mass spectrometry (MS) with higher order mass spectrometric scanning functions was utilized for localization of dosed drug or metabolite in tissue. Laser capture microscopy (LCM) was used to obtain related samples from tissue for analyses by standard MALDI-MS and HPLC-MS. In a toxicology study, rats were administered with a high dosage of a prodrug for 2 weeks. Birefringent microcrystalline material (10-25 microm) was observed in histopathologic formalin-fixed tissue samples. Direct analysis by IMS provided the identity of material in the microcrystals as circulating active drug while maintaining spatial orientation. Complementary data from visual cross-polarized light microscopy as well as standard MALDI-MS and HPLC-MS experiments on LCM samples validated the qualitative results obtained by IMS. Furthermore, the HPLC-MS analysis on the LCM samples afforded a semi-quantitative assessment of the crystalline material in the tissue samples. IMS by MALDI ion trap MS proved sensitive

  15. MoS_2/Ag nanohybrid: A novel matrix with synergistic effect for small molecule drugs analysis by negative-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Zhao, Yaju; Deng, Guoqing; Liu, Xiaohui; Sun, Liang; Li, Hui; Cheng, Quan; Xi, Kai; Xu, Danke

    2016-01-01

    This paper reports a facile synthesis of molybdenum disulfide nanosheets/silver nanoparticles (MoS_2/Ag) hybrid and its use as an effective matrix in negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The nanohybrid exerts a strong synergistic effect, leading to high performance detection of small molecule analytes including amino acids, peptides, fatty acids and drugs. The enhancement of laser desorption/ionization (LDI) efficiency is largely attributed to the high surface roughness and large surface area for analyte adsorption, better dispersibility, increased thermal conductivity and enhanced UV energy absorption as compared to pure MoS_2. Moreover, both Ag nanoparticles and the edge of the MoS_2 layers function as deprotonation sites for proton capture, facilitating the charging process in negative ion mode and promoting formation of negative ions. As a result, the MoS_2/Ag nanohybrid proves to be a highly attractive matrix in MALDI-TOF MS, with desired features such as high desorption/ionization efficiency, low fragmentation interference, high salt tolerance, and no sweet-spots for mass signal. These characteristic properties allowed for simultaneous analysis of eight different drugs and quantification of acetylsalicylic acid in the spiked human serum. This work demonstrates for the first time the fabrication and application of a novel MoS_2/Ag hybrid, and provides a new platform for use in the rapid and high throughput analysis of small molecules by mass spectrometry. - Highlights: • MoS_2/Ag nanohybrid was applied as a novel matrix in negative-ion MALDI-TOF MS. • The MoS_2/Ag nanohybrid exerted synergistic effect on the detection of small molecules. • The MoS_2/Ag nanohybrid showed good signal reproducibility and low background interferences comparing to organic matrices. • MoS_2/Ag allows simultaneous analysis of multiple drugs and quantification of acetylsalicylic acid in spiked serum samples.

  16. In situ analysis of plant tissue underivatized carbohydrates and on-probe enzymatic degraded starch by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry by using carbon nanotubes as matrix.

    Gholipour, Yousef; Nonami, Hiroshi; Erra-Balsells, Rosa

    2008-12-15

    Underivatized carbohydrates of tulip bulb and leaf tissues were characterized in situ by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) by using carbon nanotubes (CNTs) as matrix. Two sample preparation methods--(i) depositing CNTs on the fresh tissue slices placed on the probe and (ii) locating semitransparent tissues on a dried layer of CNTs on the probe--were examined. Furthermore, practicability of in situ starch analysis by MALDI-TOF MS was examined by detection of glucose originated from on-probe amyloglucosidase-catalyzed degradation of starch on the tissue surface. Besides, CNTs could efficiently desorb/ionize natural mono-, di-, and oligosaccharides extracted from tulip bulb tissues as well as glucose resulting from starch enzymatic degradation in vitro. These results were compared with those obtained by in situ MALDI-TOF MS analysis of similar tissues. Positive ion mode showed superior signal reproducibility. CNTs deposited under semitransparent tissue could also desorb/ionize neutral carbohydrates, leading to nearly complete elimination of matrix cluster signals but with an increase in tissue-originated signals. Furthermore, several experiments were carried out to compare the efficiency of 2,5-dihydroxybenzoic acid, nor-harmane, alpha-cyano-4-hydroxycinnamic acid, and CNTs as matrices for MALDI of neutral carbohydrates from the intact plant tissue surface and for enzymatic tissue starch degradation; these results are discussed in brief. Among matrices studied, the lowest laser power was needed to acquire carbohydrate signals with high signal-to-noise ratio and resolution when CNTs were used.

  17. Ultrashort pulse laser technology laser sources and applications

    Schrempel, Frank; Dausinger, Friedrich

    2016-01-01

    Ultrashort laser pulses with durations in the femtosecond range up to a few picoseconds provide a unique method for precise materials processing or medical applications. Paired with the recent developments in ultrashort pulse lasers, this technology is finding its way into various application fields. The book gives a comprehensive overview of the principles and applications of ultrashort pulse lasers, especially applied to medicine and production technology. Recent advances in laser technology are discussed in detail. This covers the development of reliable and cheap low power laser sources as well as high average power ultrashort pulse lasers for large scale manufacturing. The fundamentals of laser-matter-interaction as well as processing strategies and the required system technology are discussed for these laser sources with respect to precise materials processing. Finally, different applications within medicine, measurement technology or materials processing are highlighted.

  18. Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System for Identification of Nocardia, Rhodococcus, Kocuria, Gordonia, Tsukamurella, and Listeria Species

    Lee, Tai-Fen; Du, Shin-Hei; Teng, Shih-Hua; Liao, Chun-Hsing; Sheng, Wang-Hui; Teng, Lee-Jene

    2014-01-01

    We evaluated whether the Bruker Biotyper matrix-associated laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system provides accurate species-level identifications of 147 isolates of aerobically growing Gram-positive rods (GPRs). The bacterial isolates included Nocardia (n = 74), Listeria (n = 39), Kocuria (n = 15), Rhodococcus (n = 10), Gordonia (n = 7), and Tsukamurella (n = 2) species, which had all been identified by conventional methods, molecular methods, or both. In total, 89.7% of Listeria monocytogenes, 80% of Rhodococcus species, 26.7% of Kocuria species, and 14.9% of Nocardia species (n = 11, all N. nova and N. otitidiscaviarum) were correctly identified to the species level (score values, ≥2.0). A clustering analysis of spectra generated by the Bruker Biotyper identified six clusters of Nocardia species, i.e., cluster 1 (N. cyriacigeorgica), cluster 2 (N. brasiliensis), cluster 3 (N. farcinica), cluster 4 (N. puris), cluster 5 (N. asiatica), and cluster 6 (N. beijingensis), based on the six peaks generated by ClinProTools with the genetic algorithm, i.e., m/z 2,774.477 (cluster 1), m/z 5,389.792 (cluster 2), m/z 6,505.720 (cluster 3), m/z 5,428.795 (cluster 4), m/z 6,525.326 (cluster 5), and m/z 16,085.216 (cluster 6). Two clusters of L. monocytogenes spectra were also found according to the five peaks, i.e., m/z 5,594.85, m/z 6,184.39, and m/z 11,187.31, for cluster 1 (serotype 1/2a) and m/z 5,601.21 and m/z 11,199.33 for cluster 2 (serotypes 1/2b and 4b). The Bruker Biotyper system was unable to accurately identify Nocardia (except for N. nova and N. otitidiscaviarum), Tsukamurella, or Gordonia species. Continuous expansion of the MALDI-TOF MS databases to include more GPRs is necessary. PMID:24759706

  19. Short pulse laser systems for biomedical applications

    Mitra, Kunal

    2017-01-01

    This book presents practical information on the clinical applications of short pulse laser systems and the techniques for optimizing these applications in a manner that will be relevant to a broad audience, including engineering and medical students as well as researchers, clinicians, and technicians. Short pulse laser systems are useful for both subsurface tissue imaging and laser induced thermal therapy (LITT), which hold great promise in cancer diagnostics and treatment. Such laser systems may be used alone or in combination with optically active nanoparticles specifically administered to the tissues of interest for enhanced contrast in imaging and precise heating during LITT. Mathematical and computational models of short pulse laser-tissue interactions that consider the transient radiative transport equation coupled with a bio-heat equation considering the initial transients of laser heating were developed to analyze the laser-tissue interaction during imaging and therapy. Experiments were first performe...

  20. Holograms made with a pulsed dye laser

    Fernandez-Guasti, M.; Iturbe-Castillo, D.; Silva-Perez, A.; Gil-Villegas, A.; Gonzalez-Torres, H.; Lopez-Guerrero, R.

    1989-01-01

    We report the obtention of holograms with a nitrogen pumped dye laser, whose source is inherently pulsed. We review the advantages and posibilities of holograms of moving objects which are impossible to make with CW lasers. The lasers used in these experiments were designed and built in the quantum optics laboratory at the Universidad Autonoma Metropolitana-Iztapalapa. (Author)

  1. Intense isolated attosecond pulse generation from relativistic laser plasmas using few-cycle laser pulses

    Ma, Guangjin; Dallari, William; Borot, Antonin; Tsakiris, George D.; Veisz, Laszlo; Krausz, Ferenc; Yu, Wei

    2015-01-01

    We have performed a systematic study through particle-in-cell simulations to investigate the generation of attosecond pulse from relativistic laser plasmas when laser pulse duration approaches the few-cycle regime. A significant enhancement of attosecond pulse energy has been found to depend on laser pulse duration, carrier envelope phase, and plasma scale length. Based on the results obtained in this work, the potential of attaining isolated attosecond pulses with ∼100 μJ energy for photons >16 eV using state-of-the-art laser technology appears to be within reach

  2. Assisted inhibition effect of acetylcholinesterase with n-octylphosphonic acid and application in high sensitive detection of organophosphorous pesticides by matrix-assisted laser desorption/ionization Fourier transform mass spectrometry.

    Cai, Tingting; Zhang, Li; Wang, Haoyang; Zhang, Jing; Guo, Yinlong

    2011-11-14

    A simple and practical approach to improve the sensitivity of acetylcholinesterase (AChE)-inhibited method has been developed for monitoring organophosphorous (OP) pesticide residues. In this work, matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS) was used to detect AChE activity. Due to its good salt-tolerance and low sample consumption, MALDI-FTMS facilitates rapid and high-throughput screening of OP pesticides. Here we describe a new method to obtain low detection limits via employing external reagents. Among candidate compounds, n-octylphosphonic acid (n-Octyl-PA) displays assistant effect to enhance AChE inhibition by OP pesticides. In presence of n-Octyl-PA, the percentages of AChE inhibition still kept correlation with OP pesticide concentrations. The detection limits were improved significantly even by 10(2)-10(3) folds in comparison with conventional enzyme-inhibited methods. Different detection limits of OP pesticides with different toxicities were as low as 0.005 μg L(-1) for high toxic pesticides and 0.05 μg L(-1) for low toxic pesticides. Besides, the reliability of results from this method to analyze cowpea samples had been demonstrated by liquid-chromatography tandem mass spectrometry (LC-MS/MS). The application of this commercial available assistant agent shows great promise to detect OP compounds in complicated biological matrix and broadens the mind for high sensitivity detection of OP pesticide residues in agricultural products. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Independent assessment of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) sample preparation quality: Effect of sample preparation on MALDI-MS of synthetic polymers.

    Kooijman, Pieter C; Kok, Sander; Honing, Maarten

    2017-02-28

    Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) provides detailed and in-depth information about the molecular characteristics of synthetic polymers. To obtain the most accurate results the sample preparation parameters should be chosen to suit the sample and the aim of the experiment. Because the underlying principles of MALDI are still not fully known, a priori determination of optimal sample preparation protocols is often not possible. Employing an automated sample preparation quality assessment method recently presented by us we quantified the sample preparation quality obtained using various sample preparation protocols. Six conventional matrices with and without added potassium as a cationization agent and six ionic liquid matrices (ILMs) were assessed using poly(ethylene glycol) (PEG), polytetrahydrofuran (PTHF) and poly(methyl methacrylate) (PMMA) as samples. All sample preparation protocols were scored and ranked based on predefined quality parameters and spot-to-spot repeatability. Clearly distinctive preferences were observed in matrix identity and cationization agent for PEG, PTHF and PMMA, as the addition of an excess of potassium cationization agent results in an increased score for PMMA and a contrasting matrix-dependent effect for PTHF and PEG. The addition of excess cationization agent to sample mixtures dissipates any overrepresentation of high molecular weight polymer species. Our results show reduced ionization efficiency and similar sample deposit homogeneity for all tested ILMs, compared with well-performing conventional MALDI matrices. The results published here represent a start in the unsupervised quantification of sample preparation quality for MALDI samples. This method can select the best sample preparation parameters for any synthetic polymer sample and the results can be used to formulate hypotheses on MALDI principles. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  4. Analysis of phloem protein patterns from different organs of Cucurbita maxima Duch. by matrix-assisted laser desorption/ionization time of flight mass spectroscopy combined with sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    Kehr, J; Haebel, S; Blechschmidt-Schneider, S; Willmitzer, L; Steup, M; Fisahn, J

    1999-02-01

    Sieve tubes mediate the long-distance transport of nutrients and signals between source and sink organs of plants. To detect mobile phloem proteins that are differentially distributed in source and sink organs of Cucurbita maxima, we used both one-dimensional gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Both techniques revealed that phloem protein patterns depend on the sampling site: whilst several proteins were consistently observed in all phloem samples studied others appeared to occur in a organ-specific manner. For a characterization and identification of distinct phloem polypeptides, two approaches were chosen. First, protein bands resolved by SDS-PAGE were eluted from the polyacrylamide gel and the masses of the proteins were then determined by MALDI-TOF MS. Second, proteins resolved by SDS-PAGE were subjected to proteolytic degradation and the resulting peptides were analyzed by MALDI-TOF MS: the masses of the proteolytic peptides were used for a database search. By the latter approach, three mobile phloem compounds were identified as the phloem-specific protein PP2 (D.E. Bostwick et al., 1992, The Plant Cell 4, 1539-1548) a chymotrypsin and an aspartic proteinase inhibitor. None of the other polypeptides studied corresponded to any of the protein sequences present in the database. Furthermore, MALDI-TOF MS analyses indicated that some of the mobile phloem proteins occur in a covalently modified form and that the extent of the modification depends upon the plant organ.

  5. Misidentification of Aspergillus nomius and Aspergillus tamarii as Aspergillus flavus: characterization by internal transcribed spacer, β-Tubulin, and calmodulin gene sequencing, metabolic fingerprinting, and matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Tam, Emily W T; Chen, Jonathan H K; Lau, Eunice C L; Ngan, Antonio H Y; Fung, Kitty S C; Lee, Kim-Chung; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2014-04-01

    Aspergillus nomius and Aspergillus tamarii are Aspergillus species that phenotypically resemble Aspergillus flavus. In the last decade, a number of case reports have identified A. nomius and A. tamarii as causes of human infections. In this study, using an internal transcribed spacer, β-tubulin, and calmodulin gene sequencing, only 8 of 11 clinical isolates reported as A. flavus in our clinical microbiology laboratory by phenotypic methods were identified as A. flavus. The other three isolates were A. nomius (n = 2) or A. tamarii (n = 1). The results corresponded with those of metabolic fingerprinting, in which the A. flavus, A. nomius, and A. tamarii strains were separated into three clusters based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC MS) analysis. The first two patients with A. nomius infections had invasive aspergillosis and chronic cavitary and fibrosing pulmonary and pleural aspergillosis, respectively, whereas the third patient had A. tamarii colonization of the airway. Identification of the 11 clinical isolates and three reference strains by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) showed that only six of the nine strains of A. flavus were identified correctly. None of the strains of A. nomius and A. tamarii was correctly identified. β-Tubulin or the calmodulin gene should be the gene target of choice for identifying A. flavus, A. nomius, and A. tamarii. To improve the usefulness of MALDI-TOF MS, the number of strains for each species in MALDI-TOF MS databases should be expanded to cover intraspecies variability.

  6. Misidentification of Aspergillus nomius and Aspergillus tamarii as Aspergillus flavus: Characterization by Internal Transcribed Spacer, β-Tubulin, and Calmodulin Gene Sequencing, Metabolic Fingerprinting, and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Tam, Emily W. T.; Chen, Jonathan H. K.; Lau, Eunice C. L.; Ngan, Antonio H. Y.; Fung, Kitty S. C.; Lee, Kim-Chung; Lam, Ching-Wan; Yuen, Kwok-Yung

    2014-01-01

    Aspergillus nomius and Aspergillus tamarii are Aspergillus species that phenotypically resemble Aspergillus flavus. In the last decade, a number of case reports have identified A. nomius and A. tamarii as causes of human infections. In this study, using an internal transcribed spacer, β-tubulin, and calmodulin gene sequencing, only 8 of 11 clinical isolates reported as A. flavus in our clinical microbiology laboratory by phenotypic methods were identified as A. flavus. The other three isolates were A. nomius (n = 2) or A. tamarii (n = 1). The results corresponded with those of metabolic fingerprinting, in which the A. flavus, A. nomius, and A. tamarii strains were separated into three clusters based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC MS) analysis. The first two patients with A. nomius infections had invasive aspergillosis and chronic cavitary and fibrosing pulmonary and pleural aspergillosis, respectively, whereas the third patient had A. tamarii colonization of the airway. Identification of the 11 clinical isolates and three reference strains by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) showed that only six of the nine strains of A. flavus were identified correctly. None of the strains of A. nomius and A. tamarii was correctly identified. β-Tubulin or the calmodulin gene should be the gene target of choice for identifying A. flavus, A. nomius, and A. tamarii. To improve the usefulness of MALDI-TOF MS, the number of strains for each species in MALDI-TOF MS databases should be expanded to cover intraspecies variability. PMID:24452174

  7. A peptidomic approach for monitoring and characterising peptide cyanotoxins produced in Italian lakes by matrix-assisted laser desorption/ionisation and quadrupole time-of-flight mass spectrometry.

    Ferranti, Pasquale; Nasi, Antonella; Bruno, Milena; Basile, Adriana; Serpe, Luigi; Gallo, Pasquale

    2011-05-15

    In recent years, the occurrence of cyanobacterial blooms in eutrophic freshwaters has been described all over the world, including most European countries. Blooms of cyanobacteria may produce mixtures of toxic secondary metabolites, called cyanotoxins. Among these, the most studied are microcystins, a group of cyclic heptapeptides, because of their potent hepatotoxicity and activity as tumour promoters. Other peptide cyanotoxins have been described whose structure and toxicity have not been thoroughly studied. Herein we present a peptidomic approach aimed to characterise and quantify the peptide cyanotoxins produced in two Italian lakes, Averno and Albano. The procedure was based on matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry mass spectrometry (MALDI-TOF-MS) analysis for rapid detection and profiling of the peptide mixture complexity, combined with liquid chromatography/electrospray ionisation quadrupole time-of- flight tandem mass spectrometry (LC/ESI-Q-TOF-MS/MS) which provided unambiguous structural identification of the main compounds, as well as accurate quantitative analysis of microcystins. In the case of Lake Averno, a novel variant of microcystin-RR and two novel anabaenopeptin variants (Anabaenopeptins B(1) and Anabaenopeptin F(1)), presenting homoarginine in place of the commonly found arginine, were detected and characterised. In Lake Albano, the peculiar peptide patterns in different years were compared, as an example of the potentiality of the peptidomic approach for fast screening analysis, prior to fine structural analysis and determination of cyanotoxins, which included six novel aeruginosin variants. This approach allows for wide range monitoring of cyanobacteria blooms, and to collect data for evaluating possible health risks to consumers, through the panel of the compounds produced along different years. Copyright © 2011 John Wiley & Sons, Ltd.

  8. Source-identifying biomarker ions between environmental and clinical Burkholderia pseudomallei using whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).

    Niyompanich, Suthamat; Jaresitthikunchai, Janthima; Srisanga, Kitima; Roytrakul, Sittiruk; Tungpradabkul, Sumalee

    2014-01-01

    Burkholderia pseudomallei is the causative agent of melioidosis, which is an endemic disease in Northeast Thailand and Northern Australia. Environmental reservoirs, including wet soils and muddy water, serve as the major sources for contributing bacterial infection to both humans and animals. The whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (whole-cell MALDI-TOF MS) has recently been applied as a rapid, accurate, and high-throughput tool for clinical diagnosis and microbiological research. In this present study, we employed a whole-cell MALDI-TOF MS approach for assessing its potency in clustering a total of 11 different B. pseudomallei isolates (consisting of 5 environmental and 6 clinical isolates) with respect to their origins and to further investigate the source-identifying biomarker ions belonging to each bacterial group. The cluster analysis demonstrated that six out of eleven isolates were grouped correctly to their sources. Our results revealed a total of ten source-identifying biomarker ions, which exhibited statistically significant differences in peak intensity between average environmental and clinical mass spectra using ClinProTools software. Six out of ten mass ions were assigned as environmental-identifying biomarker ions (EIBIs), including, m/z 4,056, 4,214, 5,814, 7,545, 7,895, and 8,112, whereas the remaining four mass ions were defined as clinical-identifying biomarker ions (CIBIs) consisting of m/z 3,658, 6,322, 7,035, and 7,984. Hence, our findings represented, for the first time, the source-specific biomarkers of environmental and clinical B. pseudomallei.

  9. Rapid Differentiation of Haemophilus influenzae and Haemophilus haemolyticus by Use of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry with ClinProTools Mass Spectrum Analysis.

    Chen, Jonathan H K; Cheng, Vincent C C; Wong, Chun-Pong; Wong, Sally C Y; Yam, Wing-Cheong; Yuen, Kwok-Yung

    2017-09-01

    Haemophilus influenzae is associated with severe invasive disease, while Haemophilus haemolyticus is considered part of the commensal flora in the human respiratory tract. Although the addition of a custom mass spectrum library into the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system could improve identification of these two species, the establishment of such a custom database is technically complicated and requires a large amount of resources, which most clinical laboratories cannot afford. In this study, we developed a mass spectrum analysis model with 7 mass peak biomarkers for the identification of H. influenzae and H. haemolyticus using the ClinProTools software. We evaluated the diagnostic performance of this model using 408 H. influenzae and H. haemolyticus isolates from clinical respiratory specimens from 363 hospitalized patients and compared the identification results with those obtained with the Bruker IVD MALDI Biotyper. The IVD MALDI Biotyper identified only 86.9% of H. influenzae (311/358) and 98.0% of H. haemolyticus (49/50) clinical isolates to the species level. In comparison, the ClinProTools mass spectrum model could identify 100% of H. influenzae (358/358) and H. haemolyticus (50/50) clinical strains to the species level and significantly improved the species identification rate (McNemar's test, P mass spectrometry to handle closely related bacterial species when the proprietary spectrum library failed. This approach should be useful for the differentiation of other closely related bacterial species. Copyright © 2017 American Society for Microbiology.

  10. Analysis of the differentially expressed low molecular weight peptides in human serum via an N-terminal isotope labeling technique combining nano-liquid chromatography/matrix-assisted laser desorption/ionization mass spectrometry.

    Leng, Jiapeng; Zhu, Dong; Wu, Duojiao; Zhu, Tongyu; Zhao, Ningwei; Guo, Yinlong

    2012-11-15

    Peptidomics analysis of human serum is challenging due to the low abundance of serum peptides and interference from the complex matrix. This study analyzed the differentially expressed (DE) low molecular weight peptides in human serum integrating a DMPITC-based N-terminal isotope labeling technique with nano-liquid chromatography and matrix-assisted laser desorption/ionization mass spectrometry (nano-LC/MALDI-MS). The workflow introduced a [d(6)]-4,6-dimethoxypyrimidine-2-isothiocyanate (DMPITC)-labeled mixture of aliquots from test samples as the internal standard. The spiked [d(0)]-DMPITC-labeled samples were separated by nano-LC then spotted on the MALDI target. Both quantitative and qualitative studies for serum peptides were achieved based on the isotope-labeled peaks. The DMPITC labeling technique combined with nano-LC/MALDI-MS not only minimized the errors in peptide quantitation, but also allowed convenient recognition of the labeled peptides due to the 6 Da mass difference. The data showed that the entire research procedure as well as the subsequent data analysis method were effective, reproducible, and sensitive for the analysis of DE serum peptides. This study successfully established a research model for DE serum peptides using DMPITC-based N-terminal isotope labeling and nano-LC/MALDI-MS. Application of the DMPITC-based N-terminal labeling technique is expected to provide a promising tool for the investigation of peptides in vivo, especially for the analysis of DE peptides under different biological conditions. Copyright © 2012 John Wiley & Sons, Ltd.

  11. Early identification of microorganisms in blood culture prior to the detection of a positive signal in the BACTEC FX system using matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    Wang, Ming-Cheng; Lin, Wei-Hung; Yan, Jing-Jou; Fang, Hsin-Yi; Kuo, Te-Hui; Tseng, Chin-Chung; Wu, Jiunn-Jong

    2015-08-01

    Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is a valuable method for rapid identification of blood stream infection (BSI) pathogens. Integration of MALDI-TOF MS and blood culture system can speed the identification of causative BSI microorganisms. We investigated the minimal microorganism concentrations of common BSI pathogens required for positive blood culture using BACTEC FX and for positive identification using MALDI-TOF MS. The time to detection with positive BACTEC FX and minimal incubation time with positive MALDI-TOF MS identification were determined for earlier identification of common BSI pathogens. The minimal microorganism concentrations required for positive blood culture using BACTEC FX were >10(7)-10(8) colony forming units/mL for most of the BSI pathogens. The minimal microorganism concentrations required for identification using MALDI-TOF MS were > 10(7) colony forming units/mL. Using simulated BSI models, one can obtain enough bacterial concentration from blood culture bottles for successful identification of five common Gram-positive and Gram-negative bacteria using MALDI-TOF MS 1.7-2.3 hours earlier than the usual time to detection in blood culture systems. This study provides an approach to earlier identification of BSI pathogens prior to the detection of a positive signal in the blood culture system using MALDI-TOF MS, compared to current methods. It can speed the time for identification of BSI pathogens and may have benefits of earlier therapy choice and on patient outcome. Copyright © 2013. Published by Elsevier B.V.

  12. Species level identification of coagulase negative Staphylococcus spp. from buffalo using matrix-assisted laser desorption ionization-time of flight mass spectrometry and cydB real-time quantitative PCR.

    Pizauro, Lucas J L; de Almeida, Camila C; Soltes, Glenn A; Slavic, Durda; Rossi-Junior, Oswaldo D; de Ávila, Fernando A; Zafalon, Luiz F; MacInnes, Janet I

    2017-05-01

    Incorrect identification of Staphylococcus spp. can have serious clinical and zoonotic repercussions. Accordingly, the aim of this study was to determine if matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and/or cydB real- time quantitative PCR (qPCR) could be used to accurately identify coagulase negative Staphylococcus spp. (CoNS) obtained from buffalo milk and milking environment samples. Seventy-five of 84 CoNS isolates could be identified to the species level (score value >1.99) using MALDI-TOF MS. However, as determined by cytochrome d ubiquinol oxidase subunit II (cydB) qPCR and by 16S RNA and cydB gene sequencing, 10S. agnetis strains were wrongly identified as S. hyicus by MALDI-TOF MS. In addition, 9 isolates identified by MALDI-TOF only to the genus level (score values between 1.70 and 1.99) could be identified to species by cydB qPCR. Our findings suggest that MALDI-TOF MS is a reliable method for rapid identification of S. chromogenes and S. epidermidis (species of interest both in human and veterinary medicine) and may be able to correctly identify other Staphylococcus spp. However, at present not all Staphylococcus spp. found in buffalo milk can be accurately identified by MALDI-TOF MS and for these organisms, the cydB qPCR developed in the current study may provide a reliable alternative method for rapid identification of CoNS species. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Delineation of Stenotrophomonas maltophilia isolates from cystic fibrosis patients by fatty acid methyl ester profiles and matrix-assisted laser desorption/ionization time-of-flight mass spectra using hierarchical cluster analysis and principal component analysis.

    Vidigal, Pedrina Gonçalves; Mosel, Frank; Koehling, Hedda Luise; Mueller, Karl Dieter; Buer, Jan; Rath, Peter Michael; Steinmann, Joerg

    2014-12-01

    Stenotrophomonas maltophilia is an opportunist multidrug-resistant pathogen that causes a wide range of nosocomial infections. Various cystic fibrosis (CF) centres have reported an increasing prevalence of S. maltophilia colonization/infection among patients with this disease. The purpose of this study was to assess specific fingerprints of S. maltophilia isolates from CF patients (n = 71) by investigating fatty acid methyl esters (FAMEs) through gas chromatography (GC) and highly abundant proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and to compare them with isolates obtained from intensive care unit (ICU) patients (n = 20) and the environment (n = 11). Principal component analysis (PCA) of GC-FAME patterns did not reveal a clustering corresponding to distinct CF, ICU or environmental types. Based on the peak area index, it was observed that S. maltophilia isolates from CF patients produced significantly higher amounts of fatty acids in comparison with ICU patients and the environmental isolates. Hierarchical cluster analysis (HCA) based on the MALDI-TOF MS peak profiles of S. maltophilia revealed the presence of five large clusters, suggesting a high phenotypic diversity. Although HCA of MALDI-TOF mass spectra did not result in distinct clusters predominantly composed of CF isolates, PCA revealed the presence of a distinct cluster composed of S. maltophilia isolates from CF patients. Our data suggest that S. maltophilia colonizing CF patients tend to modify not only their fatty acid patterns but also their protein patterns as a response to adaptation in the unfavourable environment of the CF lung. © 2014 The Authors.

  14. Thymol treatment of bacteria prior to matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis aids in identifying certain bacteria at the subspecies level.

    Holland, Ricky D; Wilkes, Jon G; Cooper, Willie M; Alusta, Pierre; Williams, Anna; Pearce, Bruce; Beaudoin, Michael; Buzatu, Dan

    2014-12-15

    The identification of bacteria based on mass spectra produced by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) has become routine since its introduction in 1996. The major drawback is that bacterial patterns produced by MALDI are dependent on sample preparation prior to analysis. This results in poor reproducibility in identifying bacterial types and between laboratories. The need for a more broadly applicable and useful sample handling procedure is warranted. Thymol was added to the suspension solvent of bacteria prior to MALDI analysis. The suspension solvent consisted of ethanol, water and TFA. The bacterium was added to the thymol suspension solvent and heated. An aliquot of the bacterial suspension was mixed directly with the matrix solution at a 9:1 ratio, matrix/bacteria solution, respectively. The mixture was then placed on the MALDI plate and allowed to air dry before MALDI analysis. The thymol method improved the quality of spectra and number of peaks when compared to other sample preparation procedures studied. The bacterium-identifying biomarkers assigned to four strains of E. coli were statistically 95% reproducible analyzed on three separate days. The thymol method successfully differentiated between the four E. coli strains. In addition, the thymol procedure could identify nine out of ten S. enterica serovars over a 3-day period and nine S. Typhimurium strains from the other ten serovars 90% of the time over the same period. The thymol method can identify certain bacteria at the sub-species level and yield reproducible results over time. It improves the quality of spectra by increasing the number of peaks when compared to the other sample preparation methods assessed in this study. Published in 2014. This article is a U.S. Government work and is in the public domain in the USA. Published in 2014. This article is a U.S. Government work and is in the public domain in the USA.

  15. Phospholipid Topography of Whole-Body Sections of the Anopheles stephensi Mosquito, Characterized by High-Resolution Atmospheric-Pressure Scanning Microprobe Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.

    Khalil, Saleh M; Römpp, Andreas; Pretzel, Jette; Becker, Katja; Spengler, Bernhard

    2015-11-17

    High-resolution atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) has been employed to study the molecular anatomical structure of rodent malaria vector Anopheles stephensi mosquitoes. A dedicated sample preparation method was developed which suits both, the special tissue properties of the sample and the requirements of high-resolution MALDI imaging. Embedding in 5% carboxymethylcellulose (CMC) was used to maintain the tissue integrity of the whole mosquitoes, being very soft, fragile, and difficult to handle. Individual lipid compounds, specifically representing certain cell types, tissue areas, or organs, were detected and imaged in 20 μm-thick whole-body tissue sections at a spatial resolution of 12 μm per image pixel. Mass spectrometric data and information quality were based on a mass resolution of 70,000 (at m/z 200) and a mass accuracy of better than 2 ppm in positive-ion mode on an orbital trapping mass spectrometer. A total of 67 imaged lipids were assigned by database search and, in a number of cases, identified via additional MS/MS fragmentation studies directly from tissue. This is the first MSI study at 12 μm spatial resolution of the malaria vector Anopheles. The study provides insights into the molecular anatomy of Anopheles stephensi and the distribution and localization of major classes of glycerophospholipids and sphingolipids. These data can be a basis for future experiments, investigating, e.g., the metabolism of Plasmodium-infected and -uninfected Anopheles mosquitoes.

  16. Ribosomal subunit protein typing using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification and discrimination of Aspergillus species.

    Nakamura, Sayaka; Sato, Hiroaki; Tanaka, Reiko; Kusuya, Yoko; Takahashi, Hiroki; Yaguchi, Takashi

    2017-04-26

    Accurate identification of Aspergillus species is a very important subject. Mass spectral fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is generally employed for the rapid identification of fungal isolates. However, the results are based on simple mass spectral pattern-matching, with no peak assignment and no taxonomic input. We propose here a ribosomal subunit protein (RSP) typing technique using MALDI-TOF MS for the identification and discrimination of Aspergillus species. The results are concluded to be phylogenetic in that they reflect the molecular evolution of housekeeping RSPs. The amino acid sequences of RSPs of genome-sequenced strains of Aspergillus species were first verified and compared to compile a reliable biomarker list for the identification of Aspergillus species. In this process, we revealed that many amino acid sequences of RSPs (about 10-60%, depending on strain) registered in the public protein databases needed to be corrected or newly added. The verified RSPs were allocated to RSP types based on their mass. Peak assignments of RSPs of each sample strain as observed by MALDI-TOF MS were then performed to set RSP type profiles, which were then further processed by means of cluster analysis. The resulting dendrogram based on RSP types showed a relatively good concordance with the tree based on β-tubulin gene sequences. RSP typing was able to further discriminate the strains belonging to Aspergillus section Fumigati. The RSP typing method could be applied to identify Aspergillus species, even for species within section Fumigati. The discrimination power of RSP typing appears to be comparable to conventional β-tubulin gene analysis. This method would therefore be suitable for species identification and discrimination at the strain to species level. Because RSP typing can characterize the strains within section Fumigati, this method has potential as a powerful and reliable tool in

  17. Matrix-assisted laser desorption/ionization mass spectrometric analysis of poly(3,4-ethylenedioxythiophene) in solid-state dye-sensitized solar cells: comparison of in situ photoelectrochemical polymerization in aqueous micellar and organic media.

    Zhang, Jinbao; Ellis, Hanna; Yang, Lei; Johansson, Erik M J; Boschloo, Gerrit; Vlachopoulos, Nick; Hagfeldt, Anders; Bergquist, Jonas; Shevchenko, Denys

    2015-04-07

    Solid-state dye-sensitized solar cells (sDSCs) are devoid of such issues as electrolyte evaporation or leakage and electrode corrosion, which are typical for traditional liquid electrolyte-based DSCs. Poly(3,4-ethylenedioxythiophene) (PEDOT) is one of the most popular and efficient p-type conducting polymers that are used in sDSCs as a solid-state hole-transporting material. The most convenient way to deposit this insoluble polymer into the dye-sensitized mesoporous working electrode is in situ photoelectrochemical polymerization. Apparently, the structure and the physicochemical properties of the generated conducting polymer, which determine the photovoltaic performance of the corresponding solar cell, can be significantly affected by the preparation conditions. Therefore, a simple and fast analytical method that can reveal information on polymer chain length, possible chemical modifications, and impurities is strongly required for the rapid development of efficient solar energy-converting devices. In this contribution, we applied matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for the analysis of PEDOT directly on sDSCs. It was found that the PEDOT generated in aqueous micellar medium possesses relatively shorter polymeric chains than the PEDOT deposited from an organic medium. Furthermore, the micellar electrolyte promotes a transformation of one of the thiophene terminal units to thiophenone. The introduction of a carbonyl group into the PEDOT molecule impedes the growth of the polymer chain and reduces the conductivity of the final polymer film. Both the simplicity of sample preparation (only application of the organic matrix onto the solar cell is needed) and the rapidity of analysis hold the promise of making MALDI MS an essential tool for the physicochemical characterization of conducting polymer-based sDSCs.

  18. A rapid matrix-assisted laser desorption ionization-time of flight mass spectrometry-based method for single-plasmid tracking in an outbreak of carbapenem-resistant Enterobacteriaceae.

    Lau, Anna F; Wang, Honghui; Weingarten, Rebecca A; Drake, Steven K; Suffredini, Anthony F; Garfield, Mark K; Chen, Yong; Gucek, Marjan; Youn, Jung-Ho; Stock, Frida; Tso, Hanna; DeLeo, Jim; Cimino, James J; Frank, Karen M; Dekker, John P

    2014-08-01

    Carbapenem-resistant Enterobacteriaceae (CRE) have spread globally and represent a serious and growing threat to public health. Rapid methods for tracking plasmids carrying carbapenemase genes could greatly benefit infection control efforts. Here, we demonstrate that real-time, direct tracking of a single plasmid in a bacterial strain responsible for an outbreak is possible using a commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system. In this case, we retrospectively tracked the bla(KPC) carbapenemase gene-bearing pKpQIL plasmid responsible for a CRE outbreak that occurred at the NIH Clinical Center in 2011. An ∼ 11,109-Da MS peak corresponding to a gene product of the bla(KPC) pKpQIL plasmid was identified and characterized using a combination of proteomics and molecular techniques. This plasmid peak was present in spectra from retrospectively analyzed K. pneumoniae outbreak isolates, concordant with results from whole-genome sequencing, and absent from a diverse control set of bla(KPC)-negative clinical Enterobacteriaceae isolates. Notably, the gene characterized here is located adjacent to the bla(KPC) Tn4401 transposon on the pKpQIL plasmid. Sequence analysis demonstrates the presence of this gene in other bla(KPC) Tn4401-containing plasmids and suggests that this signature MS peak may be useful in tracking other plasmids conferring carbapenem resistance. Plasmid identification using this MALDI-TOF MS method was accomplished in as little as 10 min from isolated colonies and 30 min from positive (spiked) blood cultures, demonstrating the potential clinical utility for real-time plasmid tracking in an outbreak. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  19. Direct Identification and Antimicrobial Susceptibility Testing of Bacteria From Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry and the Vitek 2 System.

    Jo, Sung Jin; Park, Kang Gyun; Han, Kyungja; Park, Dong Jin; Park, Yeon-Joon

    2016-03-01

    We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMérieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.

  20. A Rapid Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry-Based Method for Single-Plasmid Tracking in an Outbreak of Carbapenem-Resistant Enterobacteriaceae

    Lau, Anna F.; Wang, Honghui; Weingarten, Rebecca A.; Drake, Steven K.; Suffredini, Anthony F.; Garfield, Mark K.; Chen, Yong; Gucek, Marjan; Youn, Jung-Ho; Stock, Frida; Tso, Hanna; DeLeo, Jim; Cimino, James J.; Frank, Karen M.

    2014-01-01

    Carbapenem-resistant Enterobacteriaceae (CRE) have spread globally and represent a serious and growing threat to public health. Rapid methods for tracking plasmids carrying carbapenemase genes could greatly benefit infection control efforts. Here, we demonstrate that real-time, direct tracking of a single plasmid in a bacterial strain responsible for an outbreak is possible using a commercial matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system. In this case, we retrospectively tracked the blaKPC carbapenemase gene-bearing pKpQIL plasmid responsible for a CRE outbreak that occurred at the NIH Clinical Center in 2011. An ∼11,109-Da MS peak corresponding to a gene product of the blaKPC pKpQIL plasmid was identified and characterized using a combination of proteomics and molecular techniques. This plasmid peak was present in spectra from retrospectively analyzed K. pneumoniae outbreak isolates, concordant with results from whole-genome sequencing, and absent from a diverse control set of blaKPC-negative clinical Enterobacteriaceae isolates. Notably, the gene characterized here is located adjacent to the blaKPC Tn4401 transposon on the pKpQIL plasmid. Sequence analysis demonstrates the presence of this gene in other blaKPC Tn4401-containing plasmids and suggests that this signature MS peak may be useful in tracking other plasmids conferring carbapenem resistance. Plasmid identification using this MALDI-TOF MS method was accomplished in as little as 10 min from isolated colonies and 30 min from positive (spiked) blood cultures, demonstrating the potential clinical utility for real-time plasmid tracking in an outbreak. PMID:24850353

  1. Direct bacterial identification from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry: A systematic review and meta-analysis.

    Ruiz-Aragón, Jesús; Ballestero-Téllez, Mónica; Gutiérrez-Gutiérrez, Belén; de Cueto, Marina; Rodríguez-Baño, Jesús; Pascual, Álvaro

    2017-10-27

    The rapid identification of bacteraemia-causing pathogens could assist clinicians in the timely prescription of targeted therapy, thereby reducing the morbidity and mortality of this infection. In recent years, numerous techniques that rapidly and directly identify positive blood cultures have been marketed, with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) being one of the most commonly used. The aim of this systematic review and meta-analysis was to evaluate the accuracy of MALDI-TOF (Bruker ® ) for the direct identification of positive blood culture bottles. A meta-analysis was performed to summarize the results of the 32 studies evaluated. The overall quality of the studies was moderate. For Gram-positive bacteria, overall rates of correct identification of the species ranged from 0.17 to 0.98, with a cumulative rate (random-effects model) of 0.72 (95% CI: 0.64-0.80). For Gram-negative bacteria, correct identification rates ranged from 0.66 to 1.00, with a cumulative effect of 0.92 (95% CI: 0.88-0.95). For Enterobacteriaceae, the rate was 0.96 (95% CI: 0.94-0.97). MALDI-TOF mass spectrometry shows high accuracy for the correct identification of Gram-negative bacteria, particularly Enterobacteriaceae, directly from positive blood culture bottles, and moderate accuracy for the identification of Gram-positive bacteria (low for some species). Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  2. Identification of blood culture isolates directly from positive blood cultures by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and a commercial extraction system: analysis of performance, cost, and turnaround time.

    Lagacé-Wiens, Philippe R S; Adam, Heather J; Karlowsky, James A; Nichol, Kimberly A; Pang, Paulette F; Guenther, Jodi; Webb, Amanda A; Miller, Crystal; Alfa, Michelle J

    2012-10-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry represents a revolution in the rapid identification of bacterial and fungal pathogens in the clinical microbiology laboratory. Recently, MALDI-TOF has been applied directly to positive blood culture bottles for the rapid identification of pathogens, leading to reductions in turnaround time and potentially beneficial patient impacts. The development of a commercially available extraction kit (Bruker Sepsityper) for use with the Bruker MALDI BioTyper has facilitated the processing required for identification of pathogens directly from positive from blood cultures. We report the results of an evaluation of the accuracy, cost, and turnaround time of this method for 61 positive monomicrobial and 2 polymicrobial cultures representing 26 species. The Bruker MALDI BioTyper with the Sepsityper gave a valid (score, >1.7) identification for 85.2% of positive blood cultures with no misidentifications. The mean reduction in turnaround time to identification was 34.3 h (P MALDI-TOF was used for all blood cultures and 26.5 h in a more practical setting where conventional identification or identification from subcultures was required for isolates that could not be directly identified by MALDI-TOF. Implementation of a MALDI-TOF-based identification system for direct identification of pathogens from blood cultures is expected to be associated with a marginal increase in operating costs for most laboratories. However, the use of MALDI-TOF for direct identification is accurate and should result in reduced turnaround time to identification.

  3. Bacterial rapid identification with matrix assisted laser desorption/ionization time-of-flight mass spectrometry: development of an 'in-house method' and comparison with Bruker Sepsityper(®) kit.

    Frédéric Ric, S; Antoine, M; Bodson, A; Lissoir, B

    2015-10-01

    The objective of this study was to compare an in-house matrix-assisted laser desorption ionization with time of flight (MALDI-TOF) method and a commercial MALDI-TOF kit (Sepsityper(®) kit) for direct bacterial identification in positive blood cultures. We also evaluated the time saved and the cost associated with the rapid identification techniques. We used the BACTEC(®) automated system for detecting positive blood cultures. Direct identification using Sepsityper kit and the in-house method were compared with conventional identification by MALDI-TOF using pure bacterial culture on the solid phase. We also evaluated different cut-off scores for rapid bacterial identification. In total, 127 positive blood vials were selected. The rate of rapid identification with the MALDI Sepsityper kit was 25.2% with the standard cut-off and 33.9% with the enlarged cut-off, while the results for the in-house method were 44.1 and 61.4%, respectively. Error rates with the enlarged cut-off were 6.98 (n = 3) and 2.56% (n = 2) for Sepsityper and the in-house method, respectively. Identification rates were higher for gram-negative bacteria. Direct bacterial identification succeeded in supplying rapid identification of the causative organism in cases of sepsis. The time taken to obtain a result was nearly 24  hours shorter for the direct bacterial identification methods than for conventional MALDI-TOF on solid phase culture. Compared with the Sepsityper kit, the in-house method offered better results and fewer errors, was more cost-effective and easier to use.

  4. Expression of Raf kinase inhibitor protein in human hepatoma tissues by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight methods.

    Tsao, D A; Shiau, Y F; Tseng, C S; Chang, H R

    2016-01-01

    Hepatocellular carcinoma (HCC) is the most common malignant liver tumor. To reduce the mortality and improve the effectiveness of therapy, it is important to search for changes in tumor-specific biomarkers whose function may involve in disease progression and which may be useful as potential therapeutic targets. Materials and Mehtods: In this study, we use two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to observe proteome alterations of 12 tissue pairs isolated from HCC patients: Normal and tumorous tissue. Comparing the tissue types with each other, 40 protein spots corresponding to fifteen differentially expressed between normal and cancer part of HCC patients. Raf kinase inhibitor protein (RKIP), an inhibitor of Raf-mediated activation of mitogen-activated protein kinase/extracellular signal-regulated kinase, may play an important role in cancer metastasis and cell proliferation and migration of human hepatoma cells. RKIP may be considered as a marker for HCC, because its expression level changes considerably in HCC compared with normal tissue. In addition, we used the methods of Western blotting and real time-polymerase chain reaction to analysis the protein expression and gene expression of RKIP. The result showed RKIP protein and gene expression in tumor part liver tissues of HCC patient is lower than peritumorous non-neoplastic liver tissue of the corresponding HCC samples. These results strongly suggest that RKIP may be considered to be a marker for HCC and RKIP are down-regulated in liver cancer cell.

  5. The Technical and Biological Reproducibility of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Based Typing: Employment of Bioinformatics in a Multicenter Study.

    Oberle, Michael; Wohlwend, Nadia; Jonas, Daniel; Maurer, Florian P; Jost, Geraldine; Tschudin-Sutter, Sarah; Vranckx, Katleen; Egli, Adrian

    2016-01-01

    The technical, biological, and inter-center reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS) typing data has not yet been explored. The aim of this study is to compare typing data from multiple centers employing bioinformatics using bacterial strains from two past outbreaks and non-related strains. Participants received twelve extended spectrum betalactamase-producing E. coli isolates and followed the same standard operating procedure (SOP) including a full-protein extraction protocol. All laboratories provided visually read spectra via flexAnalysis (Bruker, Germany). Raw data from each laboratory allowed calculating the technical and biological reproducibility between centers using BioNumerics (Applied Maths NV, Belgium). Technical and biological reproducibility ranged between 96.8-99.4% and 47.6-94.4%, respectively. The inter-center reproducibility showed a comparable clustering among identical isolates. Principal component analysis indicated a higher tendency to cluster within the same center. Therefore, we used a discriminant analysis, which completely separated the clusters. Next, we defined a reference center and performed a statistical analysis to identify specific peaks to identify the outbreak clusters. Finally, we used a classifier algorithm and a linear support vector machine on the determined peaks as classifier. A validation showed that within the set of the reference center, the identification of the cluster was 100% correct with a large contrast between the score with the correct cluster and the next best scoring cluster. Based on the sufficient technical and biological reproducibility of MALDI-TOF MS based spectra, detection of specific clusters is possible from spectra obtained from different centers. However, we believe that a shared SOP and a bioinformatics approach are required to make the analysis robust and reliable.

  6. The Technical and Biological Reproducibility of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS Based Typing: Employment of Bioinformatics in a Multicenter Study.

    Michael Oberle

    Full Text Available The technical, biological, and inter-center reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS typing data has not yet been explored. The aim of this study is to compare typing data from multiple centers employing bioinformatics using bacterial strains from two past outbreaks and non-related strains.Participants received twelve extended spectrum betalactamase-producing E. coli isolates and followed the same standard operating procedure (SOP including a full-protein extraction protocol. All laboratories provided visually read spectra via flexAnalysis (Bruker, Germany. Raw data from each laboratory allowed calculating the technical and biological reproducibility between centers using BioNumerics (Applied Maths NV, Belgium.Technical and biological reproducibility ranged between 96.8-99.4% and 47.6-94.4%, respectively. The inter-center reproducibility showed a comparable clustering among identical isolates. Principal component analysis indicated a higher tendency to cluster within the same center. Therefore, we used a discriminant analysis, which completely separated the clusters. Next, we defined a reference center and performed a statistical analysis to identify specific peaks to identify the outbreak clusters. Finally, we used a classifier algorithm and a linear support vector machine on the determined peaks as classifier. A validation showed that within the set of the reference center, the identification of the cluster was 100% correct with a large contrast between the score with the correct cluster and the next best scoring cluster.Based on the sufficient technical and biological reproducibility of MALDI-TOF MS based spectra, detection of specific clusters is possible from spectra obtained from different centers. However, we believe that a shared SOP and a bioinformatics approach are required to make the analysis robust and reliable.

  7. Application of targeted quantitative proteomics analysis in human cerebrospinal fluid using a liquid chromatography matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometer (LC MALDI TOF/TOF) platform.

    Pan, Sheng; Rush, John; Peskind, Elaine R; Galasko, Douglas; Chung, Kathryn; Quinn, Joseph; Jankovic, Joseph; Leverenz, James B; Zabetian, Cyrus; Pan, Catherine; Wang, Yan; Oh, Jung Hun; Gao, Jean; Zhang, Jianpeng; Montine, Thomas; Zhang, Jing

    2008-02-01

    Targeted quantitative proteomics by mass spectrometry aims to selectively detect one or a panel of peptides/proteins in a complex sample and is particularly appealing for novel biomarker verification/validation because it does not require specific antibodies. Here, we demonstrated the application of targeted quantitative proteomics in searching, identifying, and quantifying selected peptides in human cerebrospinal spinal fluid (CSF) using a matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometer (MALDI TOF/TOF)-based platform. The approach involved two major components: the use of isotopic-labeled synthetic peptides as references for targeted identification and quantification and a highly selective mass spectrometric analysis based on the unique characteristics of the MALDI instrument. The platform provides high confidence for targeted peptide detection in a complex system and can potentially be developed into a high-throughput system. Using the liquid chromatography (LC) MALDI TOF/TOF platform and the complementary identification strategy, we were able to selectively identify and quantify a panel of targeted peptides in the whole proteome of CSF without prior depletion of abundant proteins. The effectiveness and robustness of the approach associated with different sample complexity, sample preparation strategies, as well as mass spectrometric quantification were evaluated. Other issues related to chromatography separation and the feasibility for high-throughput analysis were also discussed. Finally, we applied targeted quantitative proteomics to analyze a subset of previously identified candidate markers in CSF samples of patients with Parkinson's disease (PD) at different stages and Alzheimer's disease (AD) along with normal controls.

  8. Yersinia enterocolitica in Diagnostic Fecal Samples from European Dogs and Cats: Identification by Fourier Transform Infrared Spectroscopy and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Stamm, Ivonne; Hailer, Mandy; Depner, Barbara; Kopp, Peter A.

    2013-01-01

    Yersinia enterocolitica is the main cause of yersiniosis in Europe, one of the five main bacterial gastrointestinal diseases of humans. Beside pigs, companion animals, especially dogs and cats, were repeatedly discussed in the past as a possible source of pathogenic Y. enterocolitica. To investigate the presence and types of Y. enterocolitica in companion animals, a total of 4,325 diagnostic fecal samples from dogs and 2,624 samples from cats were tested. The isolates obtained were differentiated by using matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and Fourier transform infrared spectroscopy (FT-IR). Isolated Y. enterocolitica strains were bioserotyped. The detection of the ail gene by PCR and confirmation by FT-IR were used as a pathogenicity marker. Y. enterocolitica strains were isolated from 198 (4.6%) of the dog and 8 (0.3%) of the cat fecal samples investigated. One hundred seventy-nine isolates from dogs were analyzed in detail. The virulence factor Ail was detected in 91.6% of isolates. Isolates of biotype 4 (54.7%) and, to a lesser extent, biotypes 2 (23.5%), 3 (11.2%), and 5 (2.2%) were detected. The remaining 8.4% of strains belonged to the ail-negative biotype 1A. All 7 isolates from cats that were investigated in detail were ail positive. These results indicate that companion animals could be a relevant reservoir for a broad range of presumptively human-pathogenic Y. enterocolitica types. MALDI-TOF MS and FT-IR proved to be valuable methods for the rapid identification of Y. enterocolitica, especially in regard to the large number of samples that were investigated in a short time frame. PMID:23284028

  9. Yersinia enterocolitica in diagnostic fecal samples from European dogs and cats: identification by fourier transform infrared spectroscopy and matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Stamm, Ivonne; Hailer, Mandy; Depner, Barbara; Kopp, Peter A; Rau, Jörg

    2013-03-01

    Yersinia enterocolitica is the main cause of yersiniosis in Europe, one of the five main bacterial gastrointestinal diseases of humans. Beside pigs, companion animals, especially dogs and cats, were repeatedly discussed in the past as a possible source of pathogenic Y. enterocolitica. To investigate the presence and types of Y. enterocolitica in companion animals, a total of 4,325 diagnostic fecal samples from dogs and 2,624 samples from cats were tested. The isolates obtained were differentiated by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Fourier transform infrared spectroscopy (FT-IR). Isolated Y. enterocolitica strains were bioserotyped. The detection of the ail gene by PCR and confirmation by FT-IR were used as a pathogenicity marker. Y. enterocolitica strains were isolated from 198 (4.6%) of the dog and 8 (0.3%) of the cat fecal samples investigated. One hundred seventy-nine isolates from dogs were analyzed in detail. The virulence factor Ail was detected in 91.6% of isolates. Isolates of biotype 4 (54.7%) and, to a lesser extent, biotypes 2 (23.5%), 3 (11.2%), and 5 (2.2%) were detected. The remaining 8.4% of strains belonged to the ail-negative biotype 1A. All 7 isolates from cats that were investigated in detail were ail positive. These results indicate that companion animals could be a relevant reservoir for a broad range of presumptively human-pathogenic Y. enterocolitica types. MALDI-TOF MS and FT-IR proved to be valuable methods for the rapid identification of Y. enterocolitica, especially in regard to the large number of samples that were investigated in a short time frame.

  10. Rapid and easy detection of low-level resistance to vancomycin in methicillin-resistant Staphylococcus aureus by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Asakura, Kota; Azechi, Takuya; Sasano, Hiroshi; Matsui, Hidehito; Hanaki, Hideaki; Miyazaki, Motoyasu; Takata, Tohru; Sekine, Miwa; Takaku, Tomoiku; Ochiai, Tomonori; Komatsu, Norio; Shibayama, Keigo; Katayama, Yuki; Yahara, Koji

    2018-01-01

    Vancomycin-intermediately resistant Staphylococcus aureus (VISA) and heterogeneous VISA (hVISA) are associated with treatment failure. hVISA contains only a subpopulation of cells with increased minimal inhibitory concentrations, and its detection is problematic because it is classified as vancomycin-susceptible by standard susceptibility testing and the gold-standard method for its detection is impractical in clinical microbiology laboratories. Recently, a research group developed a machine-learning classifier to distinguish VISA and hVISA from vancomycin-susceptible S. aureus (VSSA) according to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) data. Nonetheless, the sensitivity of hVISA classification was found to be 76%, and the program was not completely automated with a graphical user interface. Here, we developed a more accurate machine-learning classifier for discrimination of hVISA from VSSA and VISA among MRSA isolates in Japanese hospitals by means of MALDI-TOF MS data. The classifier showed 99% sensitivity of hVISA classification. Furthermore, we clarified the procedures for preparing samples and obtaining MALDI-TOF MS data and developed all-in-one software, hVISA Classifier, with a graphical user interface that automates the classification and is easy for medical workers to use; it is publicly available at https://github.com/bioprojects/hVISAclassifier. This system is useful and practical for screening MRSA isolates for the hVISA phenotype in clinical microbiology laboratories and thus should improve treatment of MRSA infections.

  11. Microscale Solubility Measurements of Matrix-Assisted Laser Desorption-Ionization (MALDI) Matrices Using Attenuated Total Reflection (ATR) Fourier Transform Infrared Spectroscopy (FT-IR) Coupled with Partial Least Squares (PLS) Analysis.

    Gorre, Elsa; Owens, Kevin G

    2016-11-01

    In this work an attenuated total reflection Fourier transform infrared (FT-IR) absorption based method is used to measure the solubility of two matrix-assisted laser desorption-ionization (MALDI) matrices in a few pure solvents and mixtures of acetonitrile and water using low microliter amounts of solution. Results from a method that averages the values obtained from multiple calibration curves created by manual peak picking are compared to those predicted using a partial least squares (PLS) chemometrics approach. The PLS method provided solubility values that were in good agreement with the manual method with significantly greater ease of analysis. As a test, the solubility of adipic acid in acetone was measured using the two methods of analysis, and the values are in good agreement with solubility values reported in literature. The solubilities of the MALDI matrices α-cyano-4-hydroxy cinnamic acid (CHCA) and sinapinic acid (SA) were measured in a series of mixtures made from acetonitrile (ACN) and water; surprisingly, the results show a highly nonlinear trend. While both CHCA and SA show solubility values of less than 10 mg/mL in the pure solvents, the solubility value for SA increases to 56.3 mg/mL in a 75:25 v/v ACN:water mixture. This can have a significant effect on the matrix-to-analyte ratios in the MALDI experiment when sample protocols call for preparation of a saturated solution of the matrix in the chosen solvent system. © The Author(s) 2016.

  12. Propagating Characteristics of Pulsed Laser in Rain

    Jing Guo

    2015-01-01

    Full Text Available To understand the performance of laser ranging system under the rain weather condition, we need to know the propagating characteristics of laser pulse in rain. In this paper, the absorption and attenuation coefficients were calculated based on the scattering theories in discrete stochastic media, and the propagating characteristics of laser pulse in rain were simulated and analyzed using Monte-Carlo method. Some simulation results were verified by experiments, and the simulation results are well matched with the experimental data, with the maximal deviation not less than 7.5%. The results indicated that the propagating laser beam would be attenuated and distorted due to the scattering and absorption of raindrops, and the energy attenuation and pulse shape distortion strongly depended on the laser pulse widths.

  13. LASER PROCESSING ON SINGLE CRYSTALS BY UV PULSE LASER

    龍見, 雅美; 佐々木, 徹; 高山, 恭宜

    2009-01-01

    Laser processing by using UV pulsed laser was carried out on single crystal such as sapphire and diamond in order to understand the fundamental laser processing on single crystal. The absorption edges of diamond and sapphire are longer and shorter than the wave length of UV laser, respectively. The processed regions by laser with near threshold power of processing show quite different state in each crystal.

  14. Pulsed Laser Deposition: passive and active waveguides

    Jelínek, Miroslav; Flory, F.; Escoubas, L.

    2009-01-01

    Roč. 34, č. 4 (2009), s. 438-449 ISSN 0268-1900 R&D Projects: GA ČR GA202/06/0216 Institutional research plan: CEZ:AV0Z10100522 Keywords : PLD * pulsed laser deposition * laser ablation * passive waveguides * active waveguides * waveguide laser * sensors * thin films * butane detection Subject RIV: BH - Optics, Masers, Lasers Impact factor: 0.384, year: 2009

  15. Pulsed Laser Cladding of Ni Based Powder

    Pascu, A.; Stanciu, E. M.; Croitoru, C.; Roata, I. C.; Tierean, M. H.

    2017-06-01

    The aim of this paper is to optimize the operational parameters and quality of one step Metco Inconel 718 atomized powder laser cladded tracks, deposited on AISI 316 stainless steel substrate by means of a 1064 nm high power pulsed laser, together with a Precitec cladding head manipulated by a CLOOS 7 axes robot. The optimization of parameters and cladding quality has been assessed through Taguchi interaction matrix and graphical output. The study demonstrates that very good cladded layers with low dilution and increased mechanical proprieties could be fabricated using low laser energy density by involving a pulsed laser.

  16. Superluminous laser pulse in an active medium

    Fisher, D.L.; Tajima, T.

    1993-12-01

    Physical conditions are obtained to make the propagation velocity of a laser pulse and thus the phase velocity of the excited wake be at any desired value, including that equal to or greater than the speed of light. The provision of an active-plasma laser medium with an appropriately shaped pulse allows not only replenishment of laser energy loss to the wakefield but also acceleration of the group velocity of photons. A stationary solitary solution in the accelerated frame is obtained from the model equations and simulations thereof for the laser, plasma and atoms. This approach has applications in photonics and telecommunications as well as wakefield accelerators

  17. MoS{sub 2}/Ag nanohybrid: A novel matrix with synergistic effect for small molecule drugs analysis by negative-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Zhao, Yaju, E-mail: daisy19900911@hotmail.com [State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023 (China); Deng, Guoqing, E-mail: denggqq@sina.com [Department of Polymer Science and Engineering, Nanjing University, Nanjing, 210023 (China); Liu, Xiaohui, E-mail: lcswyh@126.com [State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023 (China); Sun, Liang, E-mail: sunliang@nju.edu.cn [State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023 (China); Li, Hui, E-mail: lihui@nju.edu.cn [State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023 (China); Cheng, Quan, E-mail: quan.cheng@ucr.edu [Department of Chemistry, University of California, Riverside, CA, 92521 (United States); Xi, Kai, E-mail: xikai@nju.edu.cn [Department of Polymer Science and Engineering, Nanjing University, Nanjing, 210023 (China); Xu, Danke, E-mail: xudanke@nju.edu.cn [State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023 (China)

    2016-09-21

    This paper reports a facile synthesis of molybdenum disulfide nanosheets/silver nanoparticles (MoS{sub 2}/Ag) hybrid and its use as an effective matrix in negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The nanohybrid exerts a strong synergistic effect, leading to high performance detection of small molecule analytes including amino acids, peptides, fatty acids and drugs. The enhancement of laser desorption/ionization (LDI) efficiency is largely attributed to the high surface roughness and large surface area for analyte adsorption, better dispersibility, increased thermal conductivity and enhanced UV energy absorption as compared to pure MoS{sub 2}. Moreover, both Ag nanoparticles and the edge of the MoS{sub 2} layers function as deprotonation sites for proton capture, facilitating the charging process in negative ion mode and promoting formation of negative ions. As a result, the MoS{sub 2}/Ag nanohybrid proves to be a highly attractive matrix in MALDI-TOF MS, with desired features such as high desorption/ionization efficiency, low fragmentation interference, high salt tolerance, and no sweet-spots for mass signal. These characteristic properties allowed for simultaneous analysis of eight different drugs and quantification of acetylsalicylic acid in the spiked human serum. This work demonstrates for the first time the fabrication and application of a novel MoS{sub 2}/Ag hybrid, and provides a new platform for use in the rapid and high throughput analysis of small molecules by mass spectrometry. - Highlights: • MoS{sub 2}/Ag nanohybrid was applied as a novel matrix in negative-ion MALDI-TOF MS. • The MoS{sub 2}/Ag nanohybrid exerted synergistic effect on the detection of small molecules. • The MoS{sub 2}/Ag nanohybrid showed good signal reproducibility and low background interferences comparing to organic matrices. • MoS{sub 2}/Ag allows simultaneous analysis of multiple drugs and quantification of

  18. Metal processing with ultrashort laser pulses

    Banks, Paul S.; Felt, M. D.; Komashko, Aleksey M.; Perry, Michael D.; Rubenchik, Alexander M.; Stuart, Brent C.

    2000-08-01

    Femtosecond laser ablation has been shown to produce well-defined cuts and holes in metals with minimal heat effect to the remaining material. Ultrashort laser pulse processing shows promise as an important technique for materials processing. We will discuss the physical effects associated with processing based experimental and modeling results. Intense ultra-short laser pulse (USLP) generates high pressures and temperatures in a subsurface layer during the pulse, which can strongly modify the absorption. We carried out simulations of USLP absorption versus material and pulse parameters. The ablation rate as function of the laser parameters has been estimated. Since every laser pulse removes only a small amount of material, a practical laser processing system must have high repetition rate. We will demonstrate that planar ablation is unstable and the initially smooth crater bottom develops a corrugated pattern after many tens of shots. The corrugation growth rate, angle of incidence and the polarization of laser electric field dependence will be discussed. In the nonlinear stage, the formation of coherent structures with scales much larger than the laser wavelength was observed. Also, there appears to be a threshold fluence above which a narrow, nearly perfectly circular channel forms after a few hundred shots. Subsequent shots deepen this channel without significantly increasing its diameter. The role of light absorption in the hole walls will be discussed.

  19. Short Pulse Laser Applications Design

    Town, R.J.; Clark, D.S.; Kemp, A.J.; Lasinski, B.F.; Tabak, M.

    2008-01-01

    We are applying our recently developed, LDRD-funded computational simulation tool to optimize and develop applications of Fast Ignition (FI) for stockpile stewardship. This report summarizes the work performed during a one-year exploratory research LDRD to develop FI point designs for the National Ignition Facility (NIF). These results were sufficiently encouraging to propose successfully a strategic initiative LDRD to design and perform the definitive FI experiment on the NIF. Ignition experiments on the National Ignition Facility (NIF) will begin in 2010 using the central hot spot (CHS) approach, which relies on the simultaneous compression and ignition of a spherical fuel capsule. Unlike this approach, the fast ignition (FI) method separates fuel compression from the ignition phase. In the compression phase, a laser such as NIF is used to implode a shell either directly, or by x rays generated from the hohlraum wall, to form a compact dense (∼300 g/cm 3 ) fuel mass with an areal density of ∼3.0 g/cm 2 . To ignite such a fuel assembly requires depositing ∼20kJ into a ∼35 (micro)m spot delivered in a short time compared to the fuel disassembly time (∼20ps). This energy is delivered during the ignition phase by relativistic electrons generated by the interaction of an ultra-short high-intensity laser. The main advantages of FI over the CHS approach are higher gain, a lower ignition threshold, and a relaxation of the stringent symmetry requirements required by the CHS approach. There is worldwide interest in FI and its associated science. Major experimental facilities are being constructed which will enable 'proof of principle' tests of FI in integrated subignition experiments, most notably the OMEGA-EP facility at the University of Rochester's Laboratory of Laser Energetics and the FIREX facility at Osaka University in Japan. Also, scientists in the European Union have recently proposed the construction of a new FI facility, called HiPER, designed to

  20. Guiding of laser pulses in plasma waveguides created by linearly-polarized femtosecond laser pulses

    Lemos, N.; Cardoso, L.; Geada, J.; Figueira, G.; Albert, F.; Dias, J. M.

    2018-01-01

    We experimentally demonstrate that plasma waveguides produced with ultra-short laser pulses (sub-picosecond) in gas jets are capable of guiding high intensity laser pulses. This scheme has the unique ability of guiding a high-intensity laser pulse in a plasma waveguide created by the same laser system in the very simple and stable experimental setup. A hot plasma column was created by a femtosecond class laser that expands into an on-axis parabolic low density profile suitable to act as a wav...

  1. Bringing Pulsed Laser Welding into Production

    Olsen, Flemmming Ove

    1996-01-01

    In this paper, some research and develop-ment activities within pulsed laser welding technology at the Tech-nical University of Denmark will be described. The laser group at the Insti-tute for Manufacturing Technology has nearly 20 years of experience in laser materials process-ing. Inter......-nationally the group is mostly known for its contri-butions to the development of the laser cutting process, but further it has been active within laser welding, both in assisting industry in bringing laser welding into production in several cases and in performing fundamental R & D. In this paper some research...... activities concerning the weldability of high alloyed austenitic stainless steels for mass production industry applying industrial lasers for fine welding will be described. Studies on hot cracking sensitivity of high alloyed austenitic stainless steel applying both ND-YAG-lasers and CO2-lasers has been...

  2. Survey on modern pulsed high power lasers

    Witte, K.J.

    1985-01-01

    The requirements to be met by lasers for particle acceleration are partially similar to those already known for fusion lasers. The power level wanted in both caes is up to 100 TW or even more. The pulse durations favourable for laser accelerators are in the range from 1 ps to 1000 ps whereas fusion lasers require several ns. The energy range for laser accelerators is thus correspondingly smaller than that for fusion lasers: 1-100 kJ versus several 100 kJ. The design criteria of lasers meeting the requirements are discussed in the following. The CO 2 , iodine, Nd:glass and excimer lasers are treated in detail. The high repetition rate aspect will not be particularly addressed since for the present generation of lasers the wanted rates of far above 1 Hz are completely out of scope. Moreover, for the demonstration of principle these rates are not needed. (orig./HSI)

  3. Laser-pulsed relativistic electron gun

    Sherman, N.K.

    1986-01-01

    A relativistic (β ≅ 0.8) electron gun with good emittance and subnanosecond pulse duration which can be synchronized to picosecond laser pulses is being developed at NRC for use in studies of particle acceleration by lasers. Bursts of electron pulses exceeding 280 keV in energy have been extracted into air form a laser-driven vacuum photodiode. Trains of 5 ps pulses of ultraviolet UV light illuminate a magnesium cathode. Photoelectrons emitted from the cathode are accelerated in a graded electrostatic potential set up by a 360 kV Marx-generator. The UV pulses are obtained by doubling the frequency of a 606 nm dye laser modelocked at 160 MHz. Electron energies were measured by residual range in an echelon of Al foils. Total charge per burst was measured by picoammeter. Time structure of the bursts has been examined with plastic scintillator and a fast photomultiplier. Tests on a low voltage photodiode achieved a current density of 180 A/cm/sup 2/ from an Mg cathode, with quantum efficiency of 2.4 x 10/sup -6/ electron per UV photon. The brevity and intensity of the laser pulses cause the electric charge collected per pulse to increase linearly with bias voltage rather than according to the Langmuir-Child law. Gun emittance is about 150 mm-msr and beam brightness is about 1A/cm/sup 2/-sr. Estimated duration of individual electron pulses of a burst is about 400 ps with instantaneous current of about 0.1 mA. Energy spread within one pulse is expected to be about 15%. This gun has the potential to be a useful source of relativistic electrons for laser acceleration studies

  4. A pulsed laser polarization monitor for PEP

    Prescott, C.

    1975-01-01

    Back scattered circularly polarized laser photons are considered as a monitor for electron beam polarization. The up-down asymmetry of up to 10 percent can be measured using a wire ionization chamber with submillimeter resolution. With a pulsed laser backgrounds are to expected to be large

  5. Electromagnetically induced transparency with broadband laser pulses

    Yavuz, D. D.

    2007-01-01

    We suggest a scheme to slow and stop broadband laser pulses inside an atomic medium using electromagnetically induced transparency. Extending the suggestion of Harris et al. [Phys. Rev. Lett. 70, 552 (1993)], the key idea is to use matched Fourier components for the probe and coupling laser beams

  6. Laser Ablation of Biological Tissue Using Pulsed CO2 Laser

    Hashishin, Yuichi; Sano, Shu; Nakayama, Takeyoshi

    2010-01-01

    Laser scalpels are currently used as a form of laser treatment. However, their ablation mechanism has not been clarified because laser excision of biological tissue occurs over a short time scale. Biological tissue ablation generates sound (laser-induced sound). This study seeks to clarify the ablation mechanism. The state of the gelatin ablation was determined using a high-speed video camera and the power reduction of a He-Ne laser beam. The aim of this study was to clarify the laser ablation mechanism by observing laser excision using the high-speed video camera and monitoring the power reduction of the He-Ne laser beam. We simulated laser excision of a biological tissue by irradiating gelatin (10 wt%) with radiation from a pulsed CO 2 laser (wavelength: 10.6 μm; pulse width: 80 ns). In addition, a microphone was used to measure the laser-induced sound. The first pulse caused ablation particles to be emitted in all directions; these particles were subsequently damped so that they formed a mushroom cloud. Furthermore, water was initially evaporated by laser irradiation and then tissue was ejected.

  7. Computer control of pulsed tunable dye lasers

    Thattey, S.S.; Dongare, A.S.; Suri, B.M.; Nair, L.G.

    1992-01-01

    Pulsed tunable dye lasers are being used extensively for spectroscopic and photo-chemical experiments, and a system for acquisition and spectral analysis of a volume of data generated will be quite useful. The development of a system for wavelength tuning and control of tunable dye lasers and an acquisition system for spectral data generated in experiments with these lasers are described. With this system, it is possible to control the tuning of three lasers, and acquire data in four channels, simultaneously. It is possible to arrive at the desired dye laser wavelength with a reproducibility of ± 0.012 cm -1 , which is within the absorption width (atomic interaction) caused by pulsed dye lasers of linewidth 0.08 cm -1 . The spectroscopic data generated can be analyzed for spectral identification within absolute accuracy ± 0.012 cm -1 . (author). 6 refs., 11 figs

  8. Ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride as a novel matrix for rapid quantitative and qualitative determination of serum free fatty acids by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry

    Zhang, Yaping [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Wang, Yanmin [Department of Clinical Laboratory, Heze Municipal Hospital, Shandong (China); Guo, Shuai; Guo, Yumei; Liu, Hui [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Li, Zhili, E-mail: lizhili@ibms.pumc.edu.cn [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China)

    2013-09-10

    Graphical abstract: -- Highlights: •A novel MALDI matrix for the detection of serum free fatty acids is ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride. •Multiple point internal standard calibration curves were constructed for nine FFAs, respectively, with excellent correlation coefficients between 0.991 and 0.999. •The MALDI-MS approach was used to rapidly differentiate the patients with and without hyperglycemia and healthy controls. -- Abstract: The blood free fatty acids (FFAs), which provide energy to the cell and act as substrates in the synthesis of fats, lipoproteins, liposaccharides, and eicosanoids, involve in a number of important physiological processes. In the present study, matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) with ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride (ATNEDC) as a novel MALDI matrix in a negative ion mode was employed to directly quantify serum FFAs. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standard (IS, C{sub 17:0}) versus their corresponding intensity ratios were constructed for C{sub 14:0}, C{sub 16:1}, C{sub 16:0}, C{sub 18:0}, C{sub 18:1}, C{sub 18:2}, C{sub 18:3}, C{sub 20:4}, and C{sub 22:6}, respectively, in their mixture, with correlation coefficients between 0.991 and 0.999 and limits of detection (LODs) between 0.2 and 5.4 μM, along with the linear dynamic range of more than two orders of magnitude. The results indicate that the multiple point internal standard calibration could reduce the impact of ion suppression and improve quantification accuracy in the MALDI mode. The quantitative results of nine FFAs from 339 serum samples, including 161 healthy controls, 118 patients with hyperglycemia and 60 patients without hyperglycemia show that FFAs levels in hyperglycemic patient sera are significantly higher than those in healthy

  9. Identification of Low Molecular Weight Glutenin Alleles by Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) in Common Wheat (Triticum aestivum L.)

    Islam, Shahidul; Applebee, Marie; Appels, Rudi; Yan, Yueming; Ma, Wujun

    2015-01-01

    Low molecular weight glutenin subunits (LMW-GS) play an important role in determining dough properties and breadmaking quality. However, resolution of the currently used methodologies for analyzing LMW-GS is rather low which prevents an efficient use of genetic variations associated with these alleles in wheat breeding. The aim of the current study is to evaluate and develop a rapid, simple, and accurate method to differentiate LMW-GS alleles using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. A set of standard single LMW-GS allele lines as well as a suite of well documented wheat cultivars were collected from France, CIMMYT, and Canada. Method development and optimization were focused on protein extraction procedures and MALDI-TOF instrument settings to generate reproducible diagnostic spectrum peak profiles for each of the known wheat LMW-GS allele. Results revealed a total of 48 unique allele combinations among the studied genotypes. Characteristic MALDI-TOF peak patterns were obtained for 17 common LMW-GS alleles, including 5 (b, a or c, d, e, f), 7 (a, b, c, d or i, f, g, h) and 5 (a, b, c, d, f) patterns or alleles for the Glu-A3, Glu-B3, and Glu-D3 loci, respectively. In addition, some reproducible MALDI-TOF peak patterns were also obtained that did not match with any known alleles. The results demonstrated a high resolution and throughput nature of MALDI-TOF technology in analyzing LMW-GS alleles, which is suitable for application in wheat breeding programs in processing a large number of wheat lines with high accuracy in limited time. It also suggested that the variation of LMW-GS alleles is more abundant than what has been defined by the current nomenclature system that is mainly based on SDS-PAGE system. The MALDI-TOF technology is useful to differentiate these variations. An international joint effort may be needed to assign allele symbols to these newly identified alleles and determine their effects on end

  10. Ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride as a novel matrix for rapid quantitative and qualitative determination of serum free fatty acids by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry

    Zhang, Yaping; Wang, Yanmin; Guo, Shuai; Guo, Yumei; Liu, Hui; Li, Zhili

    2013-01-01

    Graphical abstract: -- Highlights: •A novel MALDI matrix for the detection of serum free fatty acids is ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride. •Multiple point internal standard calibration curves were constructed for nine FFAs, respectively, with excellent correlation coefficients between 0.991 and 0.999. •The MALDI-MS approach was used to rapidly differentiate the patients with and without hyperglycemia and healthy controls. -- Abstract: The blood free fatty acids (FFAs), which provide energy to the cell and act as substrates in the synthesis of fats, lipoproteins, liposaccharides, and eicosanoids, involve in a number of important physiological processes. In the present study, matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) with ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride (ATNEDC) as a novel MALDI matrix in a negative ion mode was employed to directly quantify serum FFAs. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standard (IS, C 17:0 ) versus their corresponding intensity ratios were constructed for C 14:0 , C 16:1 , C 16:0 , C 18:0 , C 18:1 , C 18:2 , C 18:3 , C 20:4 , and C 22:6 , respectively, in their mixture, with correlation coefficients between 0.991 and 0.999 and limits of detection (LODs) between 0.2 and 5.4 μM, along with the linear dynamic range of more than two orders of magnitude. The results indicate that the multiple point internal standard calibration could reduce the impact of ion suppression and improve quantification accuracy in the MALDI mode. The quantitative results of nine FFAs from 339 serum samples, including 161 healthy controls, 118 patients with hyperglycemia and 60 patients without hyperglycemia show that FFAs levels in hyperglycemic patient sera are significantly higher than those in healthy controls and patients without

  11. Matrix-assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) as a Reliable Tool to Identify Species of Catalase-negative Gram-positive Cocci not Belonging to the Streptococcus Genus.

    Almuzara, Marisa; Barberis, Claudia; Velázquez, Viviana Rojas; Ramirez, Maria Soledad; Famiglietti, Angela; Vay, Carlos

    2016-01-01

    To evaluate the performance of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) by using 190 Catalase-negative Gram-Positive Cocci (GPC) clinical isolates. All isolates were identified by conventional phenotypic tests following the proposed scheme by Ruoff and Christensen and MALDI-TOF MS (Bruker Daltonics, BD, Bremen, Germany). Two different extraction methods (direct transfer formic acid method on spot and ethanol formic acid extraction method) and different cut-offs for genus/specie level identification were used. The score cut-offs recommended by the manufacturer (≥ 2.000 for species-level, 1.700 to 1.999 for genus level and genus level, ≥ 1.700 for species-level and score genus or species. MALDI-TOF MS identification was considered correct when the result obtained from MS database agreed with the phenotypic identification result. When both methods gave discordant results, the 16S rDNA or sodA genes sequencing was considered as the gold standard identification method. The results obtained by MS concordant with genes sequencing, although discordant with conventional phenotyping, were considered correct. MS results discordant with 16S or sod A identification were considered incorrect. Using the score cut-offs recommended by the manufacturer, 97.37% and 81.05% were correctly identified to genus and species level, respectively. On the other hand, using lower cut-off scores for identification, 97.89% and 94.21% isolates were correctly identified to genus and species level respectively by MALDI-TOF MS and no significant differences between the results obtained with two extraction methods were obtained. The results obtained suggest that MALDI-TOF MS has the potential of being an accurate tool for Catalase-negative GPC identification even for those species with difficult diagnosis as Helcococcus , Abiotrophia , Granulicatella , among others. Nevertheless, expansion of the library, especially including more strains with

  12. Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health*

    Reiding, Karli R.; Ruhaak, L. Renee; Uh, Hae-Won; el Bouhaddani, Said; van den Akker, Erik B.; Plomp, Rosina; McDonnell, Liam A.; Houwing-Duistermaat, Jeanine J.; Slagboom, P. Eline; Beekman, Marian; Wuhrer, Manfred

    2017-01-01

    Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species. Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. Overall, the

  13. Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health.

    Reiding, Karli R; Ruhaak, L Renee; Uh, Hae-Won; El Bouhaddani, Said; van den Akker, Erik B; Plomp, Rosina; McDonnell, Liam A; Houwing-Duistermaat, Jeanine J; Slagboom, P Eline; Beekman, Marian; Wuhrer, Manfred

    2017-02-01

    Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex 4 HexNAc 2 to Hex 7 HexNAc 6 dHex 1 Neu5Ac 4 , as well as that of 39 glycosylation traits derived thereof. Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. Overall

  14. Determination of spatial distribution of melamine-cyanuric acid crystals in rat kidney tissue by histology and imaging matrix-assisted laser desorption/ionization quadrupole time-of-flight mass spectrometry.

    Kim, Chae-Wook; Yun, Jun-Won; Bae, Il-Hong; Lee, Joon-Seok; Kang, Hyun-Jin; Joo, Kyung-Mi; Jeong, Hye-Jin; Chung, Jin-Ho; Park, Young-Ho; Lim, Kyung-Min

    2010-01-01

    After the outbreak of acute renal failure associated with melamine-contaminated pet food, many attempts have been made to uncover the mechanism underlying the renal toxicity caused by melamine and melamine-related compounds. Using rat models, we investigated the renal crystal formation following the ingestion of a melamine-cyanuric acid mixture (M+CA, 1:1) to gain insight into the M+CA-induced renal toxicity. M+CA did not induce toxicity in precision-cut kidney slices, suggesting that M+CA does not have a direct nephrotoxicity. On the contrary, oral administration of M+CA for 3 days induced nephrotoxicity as determined by increased serum blood urea nitrogen and creatinine, reduced creatinine clearance, and enlarged kidneys in the animals treated with 50 mg/kg M+CA (melamine, 25 mg/kg, and cyanuric acid, 25 mg/kg; 2 of 10 animals) and 100 mg/kg M+CA (9 of 9 animals). While urine crystals were found in all animals treated with M+CA (25-100 mg/kg), histological examination revealed that renal crystals could be observed only in the kidneys of animals showing signs of nephrotoxicity. Remarkably, at 50 mg/kg M+CA, crystals were observed mainly in the medulla region of the kidney, while at 100 mg/kg, crystals were disseminated throughout the cortex and medulla regions. To further investigate the crystal formation by M+CA, matrix-assisted laser desorption/ionization quadrupole time-of-flight (MALDI-Q-TOF) imaging mass spectrometry detecting melamine distribution through monitoring the product ion (m/z 85, M + H) from melamine (m/z 127, M + H) was developed to directly obtain the image of melamine distribution in the kidney. The distribution image of melamine in kidney tissue confirmed that dense points of melamine were located only in the medulla region at 50 mg/kg M+CA, while at 100 mg/kg, they were disseminated widely from the cortex to medulla. These results demonstrated that M+CA ingestion could lead to crystal formation in kidney tubules along the osmotic gradient and

  15. Frequency modulation of semiconductor disk laser pulses

    Zolotovskii, I O; Korobko, D A; Okhotnikov, O G [Ulyanovsk State University, Ulyanovsk (Russian Federation)

    2015-07-31

    A numerical model is constructed for a semiconductor disk laser mode-locked by a semiconductor saturable absorber mirror (SESAM), and the effect that the phase modulation caused by gain and absorption saturation in the semiconductor has on pulse generation is examined. The results demonstrate that, in a laser cavity with sufficient second-order dispersion, alternating-sign frequency modulation of pulses can be compensated for. We also examine a model for tuning the dispersion in the cavity of a disk laser using a Gires–Tournois interferometer with limited thirdorder dispersion. (control of radiation parameters)

  16. Hybrid Pulsed Nd:YAG Laser

    Miller, Sawyer; Trujillo, Skyler; Fort Lewis College Laser Group Team

    This work concerns the novel design of an inexpensive pulsed Nd:YAG laser, consisting of a hybrid Kerr Mode Lock (KLM) and Q-switch pulse. The two pulse generation systems work independently, non simultaneously of each other, thus generating the ability for the user to easily switch between ultra-short pulse widths or large energy density pulses. Traditionally, SF57 glass has been used as the Kerr medium. In this work, novel Kerr mode-locking mediums are being investigated including: tellurite compound glass (TeO2), carbon disulfide (CS2), and chalcogenide glass. These materials have a nonlinear index of refraction orders of magnitude,(n2), larger than SF57 glass. The Q-switched pulse will utilize a Pockels cell. As the two pulse generation systems cannot be operated simultaneously, the Pockels cell and Kerr medium are attached to kinematic mounts, allowing for quick interchange between systems. Pulse widths and repetition rates will vary between the two systems. A goal of 100 picosecond pulse widths are desired for the mode-locked system. A goal of 10 nanosecond pulse widths are desired for the Q-switch system, with a desired repetition rate of 50 Hz. As designed, the laser will be useful in imaging applications.

  17. Fundamentals of laser-assisted fabrication of inorganic and organic films

    Schou, Jørgen

    2008-01-01

    The standard method for producing films by laser-assisted methods, Pulsed Laser Deposition (PLD) will be reviewed. The films considered are usually inorganic films, but also films of organic materials have been produced. Also the deposition of organic films by MAPLE (Matrix Assisted Pulsed Laser...

  18. Matrix-assisted peptide synthesis on nanoparticles.

    Khandadash, Raz; Machtey, Victoria; Weiss, Aryeh; Byk, Gerardo

    2014-09-01

    We report a new method for multistep peptide synthesis on polymeric nanoparticles of differing sizes. Polymeric nanoparticles were functionalized via their temporary embedment into a magnetic inorganic matrix that allows multistep peptide synthesis. The matrix is removed at the end of the process for obtaining nanoparticles functionalized with peptides. The matrix-assisted synthesis on nanoparticles was proved by generating various biologically relevant peptides. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.

  19. Laser-induced ion emission during polymer deposition from a flash-frozen water ice matrix

    Rodrigo, K.; Toftmann, Bo; Schou, Jørgen

    2004-01-01

    Flash-frozen water solutions of 1% weight PEG (polyethylene glycol) at -50 degreesC were used as targets at a laser wavelength of 355 nm for polymer deposition with Matrix-Assisted Pulsed Laser Evaporation (MAPLE). For medium laser fluences the transfer of PEG material to the substrate was accomp......Flash-frozen water solutions of 1% weight PEG (polyethylene glycol) at -50 degreesC were used as targets at a laser wavelength of 355 nm for polymer deposition with Matrix-Assisted Pulsed Laser Evaporation (MAPLE). For medium laser fluences the transfer of PEG material to the substrate...

  20. Pattern Laser Annealing by a Pulsed Laser

    Komiya, Yoshio; Hoh, Koichiro; Murakami, Koichi; Takahashi, Tetsuo; Tarui, Yasuo

    1981-10-01

    Preliminary experiments with contact-type pattern laser annealing were made for local polycrystallization of a-Si, local evaporation of a-Si and local formation of Ni-Si alloy. These experiments showed that the mask patterns can be replicated as annealed regions with a resolution of a few microns on substrates. To overcome shortcomings due to the contact type pattern annealing, a projection type reduction pattern laser annealing system is proposed for resistless low temperature pattern forming processes.

  1. Pulse power technology application to lasers

    Prestwich, K.R.

    1975-01-01

    Recent developments of intense relativistic electron beam accelerators and the associated pulse power technology are reviewed. The design of specific accelerators for gas laser excitation sources is discussed. A 3 MV, 800 kA, 24 ns electron beam accelerator under development for the electron beam fusion program is described along with the low jitter multichannel oil-dielectric rail switches developed for this application. This technology leads to the design of a 20 kJ, short pulse accelerator optimized gas laser excitation with radially converging electron beams. Other gas laser research requirements have led to the development of an accelerator that will produce a 0.5 MV, 20 kJ, 1 μs electron beam pulse. (auth)

  2. Pulsed laser illumination of photovoltaic cells

    Yater, Jane A.; Lowe, Roland A.; Jenkins, Phillip P.; Landis, Geoffrey A.

    1995-01-01

    In future space missions, free electron lasers (FEL) may be used to illuminate photovoltaic receivers to provide remote power. Both the radio-frequency (RF) and induction FEL produce pulsed rather than continuous output. In this work we investigate cell response to pulsed laser light which simulates the RF FEL format. The results indicate that if the pulse repetition is high, cell efficiencies are only slightly reduced compared to constant illumination at the same wavelength. The frequency response of the cells is weak, with both voltage and current outputs essentially dc in nature. Comparison with previous experiments indicates that the RF FEL pulse format yields more efficient photovoltaic conversion than does an induction FEL format.

  3. Excimer Pumped Pulsed Tunable Dye Laser

    Littman, Michael G.

    1988-06-01

    It has been recently shown and reported for the first time at this meeting, that Excimer pumping of a single-mode, short-cavity, grazing-incidence, longitudinally-pumped pulsed dye laser is feasible. In this paper the key concepts upon which this latest development is based are presented and are in a somewhat unusual form. This manuscript describes five specific dye laser examples. The five examples represent a progression from the simplest type of dye laser to the single-mode version mentioned above. The examples thus serve as a tutorial introduction to potential users of dye lasers. The article is organized into five sections or STEPS, each of which describes a different pulsed dye laser. Since the subtle points about dye lasers are best appreciated only after one actually attempts to build a working model, a PROCEDURES category is included in which details about the construction of the particular form of laser are given. As one reads through this category, think of it as looking over the shoulder of the laser builder. The NOTES category which follows is a brief but essential discussion explaining why various components and procedures are used, as well as how laser performance specifications are obtained. This subsection can he viewed as a discussion with the laser builder concerning the reasons for specific actions and choices made in the assembly of the example laser. The last category contains COMMENTS which provide additional related information pertaining to the example laser that goes beyond the earlier annotated discussion. If you like, these are the narrator's comments. At the end of the article, after the five sequential forms of the laser have been presented, there is a brief summation.

  4. Molecular dynamics study of lubricant depletion by pulsed laser heating

    Seo, Young Woo; Rosenkranz, Andreas; Talke, Frank E.

    2018-05-01

    In this study, molecular dynamics simulations were performed to numerically investigate the effect of pulsed laser heating on lubricant depletion. The maximum temperature, the lubricant depletion width, the number of evaporated lubricant beads and the number of fragmented lubricant chains were studied as a function of laser peak power, pulse duration and repetition rate. A continuous-wave laser and a square pulse laser were simulated and compared to a Gaussian pulse laser. With increasing repetition rate, pulsed laser heating was found to approach continuous-wave laser heating.

  5. Ultrashort pulsed laser technology development program

    Manke, Gerald C.

    2014-10-01

    The Department of Navy has been pursuing a technology development program for advanced, all-fiber, Ultra Short Pulsed Laser (USPL) systems via Small Business Innovative Research (SBIR) programs. Multiple topics have been published to promote and fund research that encompasses every critical component of a standard USPL system and enable the demonstration of mJ/pulse class systems with an all fiber architecture. This presentation will summarize published topics and funded programs.

  6. Ultrashort pulse laser deposition of thin films

    Perry, Michael D.; Banks, Paul S.; Stuart, Brent C.

    2002-01-01

    Short pulse PLD is a viable technique of producing high quality films with properties very close to that of crystalline diamond. The plasma generated using femtosecond lasers is composed of single atom ions with no clusters producing films with high Sp.sup.3 /Sp.sup.2 ratios. Using a high average power femtosecond laser system, the present invention dramatically increases deposition rates to up to 25 .mu.m/hr (which exceeds many CVD processes) while growing particulate-free films. In the present invention, deposition rates is a function of laser wavelength, laser fluence, laser spot size, and target/substrate separation. The relevant laser parameters are shown to ensure particulate-free growth, and characterizations of the films grown are made using several diagnostic techniques including electron energy loss spectroscopy (EELS) and Raman spectroscopy.

  7. Amorphization of silicon by femtosecond laser pulses

    Jia, Jimmy; Li Ming; Thompson, Carl V.

    2004-01-01

    We have used femtosecond laser pulses to drill submicron holes in single crystal silicon films in silicon-on-insulator structures. Cross-sectional transmission electron microscopy and energy dispersive x-ray analysis of material adjacent to the ablated holes indicates the formation of a layer of amorphous Si. This demonstrates that even when material is ablated using femtosecond pulses near the single pulse ablation threshold, sufficient heating of the surrounding material occurs to create a molten zone which solidifies so rapidly that crystallization is bypassed

  8. Organic semiconductor rubrene thin films deposited by pulsed laser evaporation of solidified solutions

    Majewska, N.; Gazda, M.; Jendrzejewski, R.; Majumdar, S.; Sawczak, M.; Śliwiński, G.

    2017-08-01

    Organic semiconductor rubrene (C42H28) belongs to most preferred spintronic materials because of the high charge carrier mobility up to 40 cm2(V·s)-1. However, the fabrication of a defect-free, polycrystalline rubrene for spintronic applications represents a difficult task. We report preparation and properties of rubrene thin films deposited by pulsed laser evaporation of solidified solutions. Samples of rubrene dissolved in aromatic solvents toluene, xylene, dichloromethane and 1,1-dichloroethane (0.23-1% wt) were cooled to temperatures in the range of 16.5-163 K and served as targets. The target ablation was provided by a pulsed 1064 nm or 266 nm laser. For films of thickness up to 100 nm deposited on Si, glass and ITO glass substrates, the Raman and AFM data show presence of the mixed crystalline and amorphous rubrene phases. Agglomerates of rubrene crystals are revealed by SEM observation too, and presence of oxide/peroxide (C42H28O2) in the films is concluded from matrix-assisted laser desorption/ionization time-of-flight spectroscopic analysis.

  9. Pulsed laser deposition of hydroxyapatite thin films

    Koch, C.F.; Johnson, S.; Kumar, D.; Jelínek, Miroslav; Chrisey, D.B.; Doraiswamy, A.; Jin, C.; Narayan, R.J.; Mihailescu, I. N.

    2007-01-01

    Roč. 27, - (2007), s. 484-494 ISSN 0928-4931 Institutional research plan: CEZ:AV0Z10100522 Keywords : hydroxyapatite * pulsed laser deposition * bioactive ceramic s Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.486, year: 2007

  10. Quantum Computation with Ultrafast Laser Pulse Shaping

    Home; Journals; Resonance – Journal of Science Education; Volume 10; Issue 6. Quantum Computation with Ultrafast Laser Pulse Shaping. Debabrata Goswami. General Article Volume 10 Issue 6 June 2005 pp 8-14. Fulltext. Click here to view fulltext PDF. Permanent link:

  11. Femtosecond laser pulse written Volume Bragg Gratings

    Richter Daniel

    2013-11-01

    Full Text Available Femtosecond laser pulses can be applied for structuring a wide range of ransparent materials. Here we want to show how to use this ability to realize Volume-Bragg-Gratings in various- mainly non-photosensitive - glasses. We will further present the characteristics of the realized gratings and a few elected applications that have been realized.

  12. Electron in the ultrashort laser pulse

    Pardy, Miroslav

    2003-01-01

    Roč. 42, č. 1 (2003), s. 99-110 ISSN 0020-7748 R&D Projects: GA MŠk LN00A100 Institutional research plan: CEZ:AV0Z2043910 Keywords : laser pulse, Volkov solution, compton effect Subject RIV: BE - The oretical Physics Impact factor: 0.476, year: 2003

  13. Investigation of laser plasma instabilities using picosecond laser pulses

    Kline, J L; Montgomery, D S; Yin, L; Flippo, K A; Shimada, T; Johnson, R P; Rose, H A; Albright, B J; Hardin, R A

    2008-01-01

    A new short-pulse version of the single-hot-spot configuration has been implemented to enhance the performance of experiments to understand Stimulated Raman Scattering. The laser pulse length was reduced from ∼200 to ∼3 ps. The reduced pulse length improves the experiment by minimizing effects such as plasma hydrodynamic evolution and ponderomotive filamentation of the interaction beam. In addition, the shortened laser pulses allow full length 2D particle-in-cell simulations of the experiments. Using the improved single-hot-spot configuration, a series of experiments to investigate kλ D scaling of SRS has been performed. Details of the experimental setup and initial results will be presented

  14. Fundamentals of laser pulse irradiation of silicon

    Rimini, E.; Baeri, P.; Russo, G.

    1985-01-01

    A computer model has been developed to describe the space and time evolution of carrier concentration, carrier energy and lattice temperature during nanosecond and picosecond laser pulse irradiation of Si single crystals. In particular the dynamic response has been evaluated for energy density of the ps laser pulse below and above the density threshold for surface melting. The obtained data allow a comparison with time-resolved reflectivity measurements reported in the literature. The available data are fitted by the computer model assuming a relaxation time for the energy transfer from the carriers to the lattice of 1 ps. The validity of the thermal model used to describe laser annealing in the nanosecond regime is assessed. (author)

  15. Phase Noise Comparision of Short Pulse Laser Systems

    S. Zhang; S. V. Benson; J. Hansknecht; D. Hardy; G. Neil; Michelle D. Shinn

    2006-12-01

    This paper describes the phase noise measurement on several different mode-locked laser systems that have completely different gain media and configurations including a multi-kW free-electron laser. We will focus on the state of the art short pulse lasers, especially the drive lasers for photocathode injectors. A comparison between the phase noise of the drive laser pulses, electron bunches and FEL pulses will also be presented.

  16. Characterization and modulation of femtosecond laser pulse

    Dorrer, Christophe

    1999-01-01

    This work brings some solutions to the characterization and control of femtosecond laser pulses. Spectral interferometry has been extensively studied; whereas this is a rather old technique, it has found new specific applications to short pulses. Several important points concerning the experimental implementation of this technique are treated. Sources of errors have been tracked and simple solutions have been found to enhance its reliability. A recently demonstrated technique for the complete characterization of short pulses has been used to characterize short pulses from Chirped Pulse Amplification Systems. This transposition of shearing interferometry to the optical frequency domain, known as Spectral Phase Interferometry for Direct Electric-field Reconstruction (SPlDER), is conceptually very interesting: for example, the inversion from the experimental data to the electric field to be characterized is completely algebraic. A reliable tool for the characterization and optimization of Chirped pulse amplification systems has been built on this principle. This is the first single-shot real-time characterization implementation of this technique. An improvement of the method has also allowed the first single-shot real-time characterization of a short pulse using a single mono-dimensional integrative detector and an algebraic inversion of the experimental data. The control of these pulses is also of prior interest. Through a collaboration with Thomson CSF-LCR, the demonstration of the use of an optically addressed light valve at the Fourier plane of a zero-dispersion line for spectral phase modulation has been made. This device allows a high-resolution control of the spectral phase of a short pulse. It is a well-adapted tool for the correction of the residual spectral phase, at the output of Chirped Pulse Amplification systems and the temporal synthesis of shaped pulses for specific experiments. (author) [fr

  17. Double nanosecond pulses generation in ytterbium fiber laser

    Veiko, V. P.; Samokhvalov, A. A., E-mail: samokhvalov.itmo@gmail.com; Yakovlev, E. B.; Zhitenev, I. Yu.; Kliushin, A. N. [Saint-Petersburg State University of Information Technologies, Mechanics and Optics, Kronverksky Pr. 49, Saint Petersburg (Russian Federation); Lednev, V. N. [Prokhorov General Physics Institute of the Russian Academy of Sciences, Vavilov Str., 38, Moscow (Russian Federation); National University of Science and Technology MISiS, Leninskyave., 4, Moscow (Russian Federation); Pershin, S. M. [Prokhorov General Physics Institute of the Russian Academy of Sciences, Vavilov Str., 38, Moscow (Russian Federation)

    2016-06-15

    Double pulse generation mode for nanosecond ytterbium fiber laser was developed. Two sequential 60-200 ns laser pulses with variable delay between them were generated by acousto-optic modulator opening with continuous diode pumping. A custom radio frequency generator was developed to produce two sequential “opening” radio pulses with a delay of 0.2–1 μs. It was demonstrated that double pulse generation did not decrease the average laser power while providing the control over the laser pulse power profile. Surprisingly, a greater peak power in the double pulse mode was observed for the second laser pulse. Laser crater studies and plasma emission measurements revealed an improved efficiency of laser ablation in the double pulse mode.

  18. Laser-supported detonation waves and pulsed laser propulsion

    Kare, J.

    1990-01-01

    A laser thermal rocket uses the energy of a large remote laser, possibly ground-based, to heat an inert propellant and generate thrust. Use of a pulsed laser allows the design of extremely simple thrusters with very high performance compared to chemical rockets. The temperatures, pressures, and fluxes involved in such thrusters (10 4 K, 10 2 atmospheres, 10 7 w/cm 2 ) typically result in the creation of laser-supported detonation (LSD) waves. The thrust cycle thus involves a complex set of transient shock phenomena, including laser-surface interactions in the ignition of the LSD wave, laser-plasma interactions in the LSD wave itself, and high-temperature nonequilibrium chemistry behind the LSD wave. The SDIO Laser Propulsion Program is investigating these phenomena as part of an overall effort to develop the technology for a low-cost Earth-to-orbit laser launch system. We will summarize the Program's approach to developing a high performance thruster, the double-pulse planar thruster, and present an overview of some results obtained to date, along with a discussion of the many research question still outstanding in this area

  19. Pulsed Power for Solid-State Lasers

    Gagnon, W; Albrecht, G; Trenholme, J; Newton, M

    2007-04-19

    Beginning in the early 1970s, a number of research and development efforts were undertaken at U.S. National Laboratories with a goal of developing high power lasers whose characteristics were suitable for investigating the feasibility of laser-driven fusion. A number of different laser systems were developed and tested at ever larger scale in pursuit of the optimum driver for laser fusion experiments. Each of these systems had associated with it a unique pulsed power option. A considerable amount of original and innovative engineering was carried out in support of these options. Ultimately, the Solid-state Laser approach was selected as the optimum driver for the application. Following this, the Laser Program at the Lawrence Livermore National Laboratory and the University of Rochester undertook aggressive efforts directed at developing the technology. In particular, at Lawrence Livermore National Laboratory, a series of laser systems beginning with the Cyclops laser and culminating in the present with the National Ignition Facility were developed and tested. As a result, a large amount of design information for solid-state laser pulsed power systems has been documented. Some of it is in the form of published papers, but most of it is buried in internal memoranda, engineering reports and LLNL annual reports. One of the goals of this book is to gather this information into a single useable format, such that it is easily accessed and understood by other engineers and physicists for use with future designs. It can also serve as a primer, which when seriously studied, makes the subsequent reading of original work and follow-up references considerably easier. While this book deals only with the solid-state laser pulsed power systems, in the bibliography we have included a representative cross section of papers and references from much of the very fine work carried out at other institutions in support of different laser approaches. Finally, in recent years, there has

  20. Laser-Induced Damage with Femtosecond Pulses

    Kafka, Kyle R. P.

    The strong electric fields of focused femtosecond laser pulses lead to non-equilibrium dynamics in materials, which, beyond a threshold intensity, causes laser-induced damage (LID). Such a strongly non-linear and non-perturbative process renders important LID observables like fluence and intensity thresholds and damage morphology (crater) extremely difficult to predict quantitatively. However, femtosecond LID carries a high degree of precision, which has been exploited in various micro/nano-machining and surface engineering applications, such as human eye surgery and super-hydrophobic surfaces. This dissertation presents an array of experimental studies which have measured the damage behavior of various materials under femtosecond irradiation. Precision experiments were performed to produce extreme spatio-temporal confinement of the femtosecond laser-solid damage interaction on monocrystalline Cu, which made possible the first successful direct-benchmarking of LID simulation with realistic damage craters. A technique was developed to produce laser-induced periodic surface structures (LIPSS) in a single pulse (typically a multi-pulse phenomenon), and was used to perform a pump-probe study which revealed asynchronous LIPSS formation on copper. Combined with 1-D calculations, this new experimental result suggests more drastic electron heating than expected. Few-cycle pulses were used to study the LID performance and morphology of commercial ultra-broadband optics, which had not been systematically studied before. With extensive surface analysis, various morphologies were observed, including LIPSS, swelling (blisters), simple craters, and even ring-shaped structures, which varied depending on the coating design, number of pulses, and air/vacuum test environment. Mechanisms leading to these morphologies are discussed, many of which are ultrafast in nature. The applied damage behavior of multi-layer dielectric mirrors was measured and compared between long pulse (150 ps

  1. Laser-driven hydrothermal process studied with excimer laser pulses

    Mariella, Raymond; Rubenchik, Alexander; Fong, Erika; Norton, Mary; Hollingsworth, William; Clarkson, James; Johnsen, Howard; Osborn, David L.

    2017-08-01

    Previously, we discovered [Mariella et al., J. Appl. Phys. 114, 014904 (2013)] that modest-fluence/modest-intensity 351-nm laser pulses, with insufficient fluence/intensity to ablate rock, mineral, or concrete samples via surface vaporization, still removed the surface material from water-submerged target samples with confinement of the removed material, and then dispersed at least some of the removed material into the water as a long-lived suspension of nanoparticles. We called this new process, which appears to include the generation of larger colorless particles, "laser-driven hydrothermal processing" (LDHP) [Mariella et al., J. Appl. Phys. 114, 014904 (2013)]. We, now, report that we have studied this process using 248-nm and 193-nm laser light on submerged concrete, quartzite, and obsidian, and, even though light at these wavelengths is more strongly absorbed than at 351 nm, we found that the overall efficiency of LDHP, in terms of the mass of the target removed per Joule of laser-pulse energy, is lower with 248-nm and 193-nm laser pulses than with 351-nm laser pulses. Given that stronger absorption creates higher peak surface temperatures for comparable laser fluence and intensity, it was surprising to observe reduced efficiencies for material removal. We also measured the nascent particle-size distributions that LDHP creates in the submerging water and found that they do not display the long tail towards larger particle sizes that we had observed when there had been a multi-week delay between experiments and the date of measuring the size distributions. This is consistent with transient dissolution of the solid surface, followed by diffusion-limited kinetics of nucleation and growth of particles from the resulting thin layer of supersaturated solution at the sample surface.

  2. Crack imaging by pulsed laser spot thermography

    Li, T; Almond, D P; Rees, D A S; Weekes, B

    2010-01-01

    A surface crack close to a spot heated by a laser beam impedes lateral heat flow and produces alterations to the shape of the thermal image of the spot that can be monitored by thermography. A full 3D simulation has been developed to simulate heat flow from a laser heated spot in the proximity of a crack. The modelling provided an understanding of the ways that different parameters affect the thermal images of laser heated spots. It also assisted in the development of an efficient image processing strategy for extracting the scanned cracks. Experimental results show that scanning pulsed laser spot thermography has considerable potential as a remote, non-contact crack imaging technique.

  3. Pulse Compression Techniques for Laser Generated Ultrasound

    Anastasi, R. F.; Madaras, E. I.

    1999-01-01

    Laser generated ultrasound for nondestructive evaluation has an optical power density limit due to rapid high heating that causes material damage. This damage threshold limits the generated ultrasound amplitude, which impacts nondestructive evaluation inspection capability. To increase ultrasound signal levels and improve the ultrasound signal-to-noise ratio without exceeding laser power limitations, it is possible to use pulse compression techniques. The approach illustrated here uses a 150mW laser-diode modulated with a pseudo-random sequence and signal correlation. Results demonstrate the successful generation of ultrasonic bulk waves in aluminum and graphite-epoxy composite materials using a modulated low-power laser diode and illustrate ultrasound bandwidth control.

  4. Pulsed laser radiation therapy of skin tumors

    Kozlov, A.P.; Moskalik, K.G.

    1980-11-15

    Radiation from a neodymium laser was used to treat 846 patients with 687 precancerous lesions or benign tumors of the skin, 516 cutaneous carcinomas, 33 recurrences of cancer, 51 melanomas, and 508 metastatic melanomas in the skin. The patients have been followed for three months to 6.5 years. No relapses have been observed during this period. Metastases to regional lymph nodes were found in five patients with skin melanoma. Pulsed laser radiation may be successfully used in the treatment of precancerous lesions and benign tumors as well as for skin carcinoma and its recurrences, and for skin melanoma. Laser radiation is more effective in the treatment of tumors inaccessible to radiation therapy and better in those cases in which surgery may have a bad cosmetic or even mutilating effect. Laser beams can be employed in conjunction with chemo- or immunotherapy.

  5. High laser-fluence deposition of organic materials in water ice matrices by ''MAPLE''

    Christensen, Bo Toftmann; Rodrigo, K.; Schou, Jørgen

    2005-01-01

    Matrix assisted pulsed laser evaporation (MAPLE) is a deposition technique for organic material. Water ice was used as a matrix for the biotechnologically important guest material, polyethylene glycol (PEG), for concentrations from 0.5 to 4 wt.%. The target was irradiated with 6 ns laser pulses...

  6. Acousto-optic replication of ultrashort laser pulses

    Yushkov, Konstantin B.; Molchanov, Vladimir Ya.; Ovchinnikov, Andrey V.; Chefonov, Oleg V.

    2017-10-01

    Precisely controlled sequences of ultrashort laser pulses are required in various scientific and engineering applications. We developed a phase-only acousto-optic pulse shaping method for replication of ultrashort laser pulses in a TW laser system. A sequence of several Fourier-transform-limited pulses is generated from a single femtosecond laser pulse by means of applying a piecewise linear phase modulation over the whole emission spectrum. Analysis demonstrates that the main factor which limits maximum delay between the pulse replicas is spectral resolution of the acousto-optic dispersive delay line used for pulse shaping. In experiments with a Cr:forsterite laser system, we obtained delays from 0.3 to 3.5 ps between two replicas of 190 fs transform-limited pulses at the central wavelength of laser emission, 1230 nm.

  7. Pulsed laser triggered high speed microfluidic switch

    Wu, Ting-Hsiang; Gao, Lanyu; Chen, Yue; Wei, Kenneth; Chiou, Pei-Yu

    2008-10-01

    We report a high-speed microfluidic switch capable of achieving a switching time of 10 μs. The switching mechanism is realized by exciting dynamic vapor bubbles with focused laser pulses in a microfluidic polydimethylsiloxane (PDMS) channel. The bubble expansion deforms the elastic PDMS channel wall and squeezes the adjacent sample channel to control its fluid and particle flows as captured by the time-resolved imaging system. A switching of polystyrene microspheres in a Y-shaped channel has also been demonstrated. This ultrafast laser triggered switching mechanism has the potential to advance the sorting speed of state-of-the-art microscale fluorescence activated cell sorting devices.

  8. Optical reprogramming with ultrashort femtosecond laser pulses

    Uchugonova, Aisada; Breunig, Hans G.; Batista, Ana; König, Karsten

    2015-03-01

    The use of sub-15 femtosecond laser pulses in stem cell research is explored with particular emphasis on the optical reprogramming of somatic cells. The reprogramming of somatic cells into induced pluripotent stem (iPS) cells can be evoked through the ectopic expression of defined transcription factors. Conventional approaches utilize retro/lenti-viruses to deliver genes/transcription factors as well as to facilitate the integration of transcription factors into that of the host genome. However, the use of viruses may result in insertional mutations caused by the random integration of genes and as a result, this may limit the use within clinical applications due to the risk of the formation of cancer. In this study, a new approach is demonstrated in realizing non-viral reprogramming through the use of ultrashort laser pulses, to introduce transcription factors into the cell so as to generate iPS cells.

  9. Brief review on pulse laser propulsion

    Yu, Haichao; Li, Hanyang; Wang, Yan; Cui, Lugui; Liu, Shuangqiang; Yang, Jun

    2018-03-01

    Pulse laser propulsion (PLP) is an advanced propulsion concept can be used across a variety of fields with a wide range of applications. PLP reflects superior payload as well as decreased launch costs in comparison with other conventional methods of producing thrust, such as chemical propulsion or electric propulsion. Numerous researchers have attempted to exploit the potential applications of PLP. This paper first reviews concepts relevant to PLP, including the propulsion modes, breakdown regimes, and propulsion efficiency; the propulsion targets for different materials with the pulse laser are then discussed in detail, including the propulsion of solid and liquid microspheres. PLP applications such as the driven microsatellite, target surface particle removal, and orbital debris removal are also discussed. Although the PLP has been applied to a variety of fields, further research is yet warranted to establish its application in the aerospace field.

  10. Laser Pulse Heating of Spherical Metal Particles

    Michael I. Tribelsky

    2011-12-01

    Full Text Available We consider the general problem of laser pulse heating of spherical metal particles with the sizes ranging from nanometers to millimeters. We employ the exact Mie solution of the diffraction problem and solve the heat-transfer equation to determine the maximum temperature rise at the particle surface as a function of optical and thermometric parameters of the problem. Primary attention is paid to the case when the thermal diffusivity of the particle is much larger than that of the environment, as it is in the case of metal particles in fluids. We show that, in this case, for any given duration of the laser pulse, the maximum temperature rise as a function of the particle size reaches a maximum at a certain finite size of the particle. We suggest simple approximate analytical expressions for this dependence, which cover the entire parameter range of the problem and agree well with direct numerical simulations.

  11. Pulsed laser damage to optical fibers

    Allison, S.W.; Gillies, G.T.; Magnuson, D.W.; Pagano, T.S.

    1985-01-01

    This paper describes some observations of pulsed laser damage to optical fibers with emphasis on a damage mode characterized as a linear fracture along the outer core of a fiber. Damage threshold data are presented which illustrate the effects of the focusing lens, end-surface preparation, and type of fiber. An explanation based on fiber-beam misalignment is given and is illustrated by a simple experiment and ray trace

  12. A ns-Pulse Laser Microthruster

    Phipps, Claude R.; Luke, James R.; Helgeson, Wesley; Johnson, Richard

    2006-01-01

    We have developed a prototype device which demonstrates the feasibility of using ns-duration laser pulses in a laser microthruster. Relative to the ms-duration thrusters which we have demonstrated in the past, this change offers the use of any target material, the use of reflection-mode target illumination, and adjustable specific impulse. Specific impulse is adjusted by varying laser intensity on target. In this way, we were able to vary specific impulse from 200s to 3,200s on gold. We used a Concepts Research, Inc. microchip laser with 170mW average optical power, 8kHz repetition rate and 20μJ pulse energy for many of the measurements. Thrust was in the 100nN - 1μN range for all the work, requiring development of an extremely sensitive, low-noise thrust stand. We will discuss the design of metallic fuel delivery systems. Ablation efficiency near 100% was observed. Results obtained on metallic fuel systems agreed with simulations. We also report time-of-flight measurements on ejected metal ions, which gave velocities up to 80km/s

  13. Liquid Atomization Induced by Pulse Laser Reflection underneath Liquid Surface

    Utsunomiya, Yuji; Kajiwara, Takashi; Nishiyama, Takashi; Nagayama, Kunihito; Kubota, Shiro; Nakahara, Motonao

    2009-05-01

    We observed a novel effect of pulse laser reflection at the interface between transparent materials with different refractive indices. The electric field intensity doubles when a laser beam is completely reflected from a material with a higher refractive index to a material with a lower index. This effect appreciably reduces pulse laser ablation threshold of transparent materials. We performed experiments to observe the entire ablation process for laser incidence on the water-air interface using pulse laser shadowgraphy with high-resolution film; the minimum laser fluence for laser ablation at the water-air interface was approximately 12-16 J/cm2. We confirmed that this laser ablation occurs only when the laser beam is incident on the water-air interface from water. Many slender liquid ligaments extend like a milk crown and seem to be atomized at the tip. Their detailed structures can be resolved only by pulse laser photography using high-resolution film.

  14. Pulsed Nd-YAG laser in endodontics

    Ragot-Roy, Brigitte; Severin, Claude; Maquin, Michel

    1994-12-01

    The purpose of this study was to establish an operative method in endodontics. The effect of a pulsed Nd:YAG laser on root canal dentin has been examined with a scanning electron microscope. Our first experimentation was to observe the impacts carried out perpendicularly to root canal surface with a 200 micrometers fiber optic in the presence of dye. Secondarily, the optical fiber was used as an endodontic instrument with black dye. The irradiation was performed after root canal preparation (15/100 file or 40/100 file) or directly into the canal. Adverse effects are observed. The results show that laser irradiation on root canal dentin surfaces induces a nonhomogeneous modified dentin layer, melted and resolidified dentin closed partially dentinal tubules. The removal of debris is not efficient enough. The laser treatment seems to be indicated only for endodontic and periapical spaces sterilization after conventional root canal preparation.

  15. Picoseconds pulse generation and pulse width determination processes of a distributed feedback dye laser

    Abdul Ghani, B.; Hammadi, M.

    2004-08-01

    A mathematical model has been developed to describe the dynamic emission of Nd-glass, distributed feedback dye laser (DFDL), and periodical grating temperature. The suggested model allows the investigation of the time behavior of Nd-glass laser and DFDL pulsed. Moreover, it allows studying the effect of the laser input parameters of Nd-glass laser on the spectral characteristics of the output DFDL pulses such as pulse width, delay time, and time separation

  16. Regimes of self-pulsing in photonic crystal Fano lasers

    Rasmussen, Thorsten Svend; Yu, Yi; Mørk, Jesper

    2017-01-01

    Laser self-pulsing was a property exclusive to macroscopic laser systems until recently, where self-pulsing laser operation was demonstrated experimentally and theoretically in a microscopic photonic crystal Fano laser [1]. We now provide a detailed theoretical analysis of the self......-pulsing mechanism and laser characteristics with numerical simulations to demonstrate the parameter dependence of the self-pulsing regime and its limitations, indicating how the design may be optimised for applications in e.g. integrated on-chip communication systems....

  17. Dye laser spectrometer for the analysis of pulsed vacuum arcs

    Hargis, P.J. Jr.; Robertson, M.M.

    1975-01-01

    A pulsed dye laser spectrometer which is used to obtain detailed single shot spectroscopic measurements of the plasma in a pulsed vacuum arc was developed. The capabilities of this spectrometer are indicated by the detection of laser induced fluorescence signals from 10 6 neutral Ti atoms in the plasma of a pulsed vacuum arc with a Ti anode. (U.S.)

  18. Drilling of Copper Using a Dual-Pulse Femtosecond Laser

    Chung-Wei Cheng

    2016-02-01

    Full Text Available The drilling of copper using a dual-pulse femtosecond laser with wavelength of 800 nm, pulse duration of 120 fs and a variable pulse separation time (0.1–150 ps is investigated theoretically. A one-dimensional two-temperature model with temperature-dependent material properties is considered, including dynamic optical properties and the thermal-physical properties. Rapid phase change and phase explosion models are incorporated to simulate the material ablation process. Numerical results show that under the same total laser fluence of 4 J/cm2, a dual-pulse femtosecond laser with a pulse separation time of 30–150 ps can increase the ablation depth, compared to the single pulse. The optimum pulse separation time is 85 ps. It is also demonstrated that a dual pulse with a suitable pulse separation time for different laser fluences can enhance the ablation rate by about 1.6 times.

  19. Wavelength stabilisation during current pulsing of tapered laser

    Jensen, Ole Bjarlin

    2009-01-01

    The use of external feedback to stabilise the frequency of a tapered laser during current pulsing is reported. Using this technique more than 20 W of peak power in 60 ns pulses from the tapered laser is obtained and owing to the external feedback, the laser is tunable in the 778-808 nm range...

  20. Detection of diamond in ore using pulsed laser Raman spectroscopy

    Lamprecht, GH

    2007-10-01

    Full Text Available is necessary for correcting for fluorescence of minerals and diamond itself. Various pulsed laser wavelengths from 266 to 1064 nm were used, as well as cw lasers for comparison. Wavelength scans of the regions of interest, indicated that pulsed lasers at 532...

  1. Theory of Self-pulsing in Photonic Crystal Fano Lasers

    Rasmussen, Thorsten Svend; Yu, Yi; Mørk, Jesper

    2017-01-01

    -dispersive Fano mirror, the laser frequency and the threshold gain. The model is based upon a combination of conventional laser rate equations and coupled-mode theory. The dynamical model is used to demonstrate how the laser has two regimes of operation, continuous-wave output and self-pulsing, and these regimes......Laser self-pulsing was a phenomenon exclusive to macroscopic lasers until recently, where self-starting laser pulsation in a microscopic photonic crystal Fano laser was reported. In this paper a theoretical model is developed to describe the Fano laser, including descriptions of the highly...

  2. Effects of laser wavelengths and pulse energy ratio on the emission enhancement in dual pulse LIBS

    Ahmed, Rizwan; Iqbal, Javed; Baig, M Aslam

    2015-01-01

    We present new studies on the effects of laser wavelengths, pulse energy ratio and interpulse delay between two laser pulses in the collinear dual pulse configuration of laser-induced breakdown spectroscopy (LIBS) on an iron sample in air using the fundamental (1064 nm) and the second harmonics (532 nm) of Nd:YAG lasers. In the dual pulse LIBS, an optimum value of interpulse delay with an appropriate combination of laser wavelengths, and laser pulse energy ratio, yields a 30 times signal intensity enhancement in the neutral iron lines as compared with single pulse LIBS. A comparison in the spatial variations of electron temperature along the axis of the plume expansion in single and double pulse LIBS has also been studied. (letter)

  3. Measurement Issues In Pulsed Laser Propulsion

    Sinko, John E.; Scharring, Stefan; Eckel, Hans-Albert; Roeser, Hans-Peter; Sasoh, Akihiro

    2010-01-01

    Various measurement techniques have been used throughout the over 40-year history of laser propulsion. Often, these approaches suffered from inconsistencies in definitions of the key parameters that define the physics of laser ablation impulse generation. Such parameters include, but are not limited to the pulse energy, spot area, imparted impulse, and ablated mass. The limits and characteristics of common measurement techniques in each of these areas will be explored as they relate to laser propulsion. The idea of establishing some standardization system for laser propulsion data is introduced in this paper, so that reported results may be considered and studied by the general community with more certain understanding of particular merits and limitations. In particular, it is the intention to propose a minimum set of requirements a literature study should meet. Some international standards for measurements are already published, but modifications or revisions of such standards may be necessary for application to laser ablation propulsion. Issues relating to development of standards will be discussed, as well as some examples of specific experimental circumstances in which standardization would have prevented misinterpretation or misuse of past data.

  4. Repetitively pulsed, double discharge TEA CO/sub 2/ laser

    Hamilton, D C; James, D J; Ramsden, S A

    1975-10-01

    The design and operation of a repetitively pulsed TEA CO/sub 2/ laser is described. Average powers of up to 400 W at a repetition frequency of 200 pulses/s have been obtained. The system has also been used to provide long pulses (over 20 ..mu..s) and tunable single axial mode pulses.

  5. Scattering of Femtosecond Laser Pulses on the Negative Hydrogen Ion

    Astapenko, V. A.; Moroz, N. N.

    2018-05-01

    Elastic scattering of ultrashort laser pulses (USLPs) on the negative hydrogen ion is considered. Results of calculations of the USLP scattering probability are presented and analyzed for pulses of two types: the corrected Gaussian pulse and wavelet pulse without carrier frequency depending on the problem parameters.

  6. Optimizing chirped laser pulse parameters for electron acceleration in vacuum

    Akhyani, Mina; Jahangiri, Fazel; Niknam, Ali Reza; Massudi, Reza, E-mail: r-massudi@sbu.ac.ir [Laser and Plasma Research Institute, Shahid Beheshti University, Tehran 1983969411 (Iran, Islamic Republic of)

    2015-11-14

    Electron dynamics in the field of a chirped linearly polarized laser pulse is investigated. Variations of electron energy gain versus chirp parameter, time duration, and initial phase of laser pulse are studied. Based on maximizing laser pulse asymmetry, a numerical optimization procedure is presented, which leads to the elimination of rapid fluctuations of gain versus the chirp parameter. Instead, a smooth variation is observed that considerably reduces the accuracy required for experimentally adjusting the chirp parameter.

  7. Development of short pulse laser pumped x-ray lasers

    Dunn, J; Osterheld, A L; Hunter, J R; Shlyaptsev, V N

    2000-01-01

    X-ray lasers have been extensively studied around the world since the first laboratory demonstration on the Novette laser at LLNL in 1984 [l]. The characteristic properties of short wavelength, high monochromaticity, collimation and coherence make x-ray lasers useful for various applications. These include demonstrations of biological imaging within the water window, interferometry of laser plasmas and radiography of laser-heated surfaces. One of the critical issues has been the high power pump required to produce the inversion. The power scaling as a function of x-ray laser wavelength follows a -k4 to law. The shortest x-ray laser wavelength of ∼ 35 (angstrom) demonstrated for Ni-like All was at the limit of Nova laser capabilities. By requiring large, high power lasers such as Nova, the shot rate and total number of shots available have limited the rapid development of x-ray lasers and applications. In fact over the last fifteen years the main thrust has been to develop more efficient, higher repetition rate x-ray lasers that can be readily scaled to shorter wavelengths. The recent state of progress in the field can be found in references. The objective of the project was to develop a soft x-ray laser (XRL) pumped by a short pulse laser of a few joules. In effect to demonstrate a robust, worlung tabletop x-ray laser at LLNL for the first time. The transient collisional scheme as proposed by Shlyaptsev et al [8, 9] was the candidate x-ray laser for study. The successful endeavour of any scientific investigation is often based upon prudent early decisions and the choice of this scheme was both sound and fruitful. It had been demonstrated very recently for Ne-like Ti at 326 A using a small tabletop laser [10] but had not yet reached its full potential. We chose this scheme for several reasons: (a) it was a collisional-type x-ray laser which has been historically the most robust; (b) it had the promise of high efficiency and low energy threshold for lasing; (c) the

  8. Pulsed laser deposition—invention or discovery?

    Venkatesan, T

    2014-01-01

    The evolution of pulsed laser deposition had been an exciting process of invention and discovery, with the development of high T c superconducting films as the main driver. It has become the method of choice in research and development for rapid prototyping of multicomponent inorganic materials for preparing a variety of thin films, heterostructures and atomically sharp interfaces, and has become an indispensable tool for advancing oxide electronics. In this paper I will give a personal account of the invention and development of this process at Bellcore/Rutgers, the opportunity, challenges and mostly the extraordinary excitement that was generated, typical of any disruptive technology. (paper)

  9. Three-dimensional laser pulse intensity diagnostic for photoinjectors

    Heng Li

    2011-11-01

    Full Text Available Minimizing the electron-beam emittance of photoinjectors is an important task for maximizing the brightness of the next-generation x-ray facilities, such as free-electron lasers and energy recovery linacs. Optimally shaped laser pulses can significantly reduce emittance. A reliable diagnostic for the laser pulse intensity is required for this purpose. We demonstrate measurement of three-dimensional spatiotemporal intensity profiles, with spatial resolution of 20  μm and temporal resolution of 130 fs. The capability is illustrated by measurements of stacked soliton pulses and pulses from a dissipative-soliton laser.

  10. Arbitrary temporal shape pulsed fiber laser based on SPGD algorithm

    Jiang, Min; Su, Rongtao; Zhang, Pengfei; Zhou, Pu

    2018-06-01

    A novel adaptive pulse shaping method for a pulsed master oscillator power amplifier fiber laser to deliver an arbitrary pulse shape is demonstrated. Numerical simulation has been performed to validate the feasibility of the scheme and provide meaningful guidance for the design of the algorithm control parameters. In the proof-of-concept experiment, information on the temporal property of the laser is exchanged and evaluated through a local area network, and the laser adjusted the parameters of the seed laser according to the monitored output of the system automatically. Various pulse shapes, including a rectangular shape, ‘M’ shape, and elliptical shape are achieved through experimental iterations.

  11. Driver circuit for pulse modulation of a semiconductor laser

    Ueki, A.

    1975-01-01

    A pulse modulation driver circuit for a semiconductor laser is disclosed which discriminates among input pulse signals composed of binary codes to detect the occurrence of a pulse having a code of ''I'' following a pulse having a code of ''0''. Detection of this pattern is used to control the driver to increase either or both the width or peak value of the pulse having a code of 1. The effect of this is to eliminate a pattern effect in the light emitted by the semiconductor laser caused by an attenuation of the population inversion in the laser. (U.S.)

  12. Channeling and stability of laser pulses in plasmas

    Sprangle, P.; Krall, J.; Esarey, E.

    1995-01-01

    A laser pulse propagating in a plasma is found to undergo a combination of hose and modulation instabilities. The coupled equations for the laser beam envelope and centroid are derived and solved for a laser pulse of finite length propagating through either a uniform plasma or preformed plasma density channel. The laser envelope equation describes the pulse self-focusing and optical guiding in plasmas and is used to analyze the self-modulation instability. The laser centroid equation describes the transverse motion of the laser pulse (hosing) in plasmas. Significant coupling between the centroid and envelope motion as well as harmonic generation in the envelope can occur. In addition, the transverse profile of the generated wake field is strongly affected by the laser hose instability. Methods to reduce the laser hose instability are demonstrated. copyright 1995 American Institute of Physics

  13. Cubic phase control of ultrashort laser pulses

    Mecseki, K.; Erdelyi, M.; Kovacs, A.P.; Szabo, G.

    2006-01-01

    Complete test of publication follows. The temporal shape of an ultrashort laser pulse may change upon propagating through a linear dispersive medium having a phase shift ψω. The change can be characterized by the Taylor-coefficients of the phase shift which are calculated around the central frequency ω 0 of the pulse. Measurements and independent control of the group delay dispersion (GDD, ψ'(ω 0 )) and the third order dispersion (TOD, ψ'(ω 0 )) are important in several research fields, particularly in the generation of ultrashort laser pulses by chirped pulse amplification (CPA) and pulse shaping for molecular control. The GDD and the TOD of an ideal pulse compressor are equal to the negative of the corresponding dispersion coefficients of the medium. However, in the case of prism-pair and grating-pair compressor is different from the ratio of the coefficients of the medium to be compensated for. Therefore it is necessary to develop so-called cubic compressors that are able to control the TOD of the pulse, yet, do not affect the GDD. In this paper a new cubic compressor setup is investigated theoretically and experimentally, which resembles the set-up proposed by White, however, we control the GDD and the TOD by the position of a birefringent, semi-cylinder crystal place around the focal point of an achromatic lens. For the evaluation of the phase shift introduced by the proposed cubic compressor, a ray tracing program was written. The program allows optimizing the compressor parameters, such as the radius of the crystal, magnification of the lens etc. Calcite was applied because it is a strong birefringent material. Calculations showed that there is a trajectory, along which shifting the crystal the TOD can be tuned independently of the GDD. The value of the TOD changed in a relatively wide range between -3.15 x 10 5 fs 3 and -1.67 x 10 5 fs 3 . Although the defocus also affects the angular dispersion of the pulse leaving the compressor, if does not exceed

  14. PHASE NOISE COMPARISON OF SHORT PULSE LASER SYSTEMS

    Shukui Zhang; Stephen Benson; John Hansknecht; David Hardy; George Neil; Michelle D. Shinn

    2006-08-27

    This paper describes phase noise measurements of several different laser systems that have completely different gain media and configurations including a multi-kW free-electron laser. We will focus on state-of-the-art short pulse lasers, especially drive lasers for photocathode injectors. Phase noise comparison of the FEL drive laser, electron beam and FEL laser output also will be presented.

  15. 25 years of pulsed laser deposition

    Lorenz, Michael; Ramachandra Rao, M. S.

    2014-01-01

    It is our pleasure to introduce this special issue appearing on the occasion of the 25th anniversary of pulsed laser deposition (PLD), which is today one of the most versatile growth techniques for oxide thin films and nanostructures. Ever since its invention, PLD has revolutionized the research on advanced functional oxides due to its ability to yield high-quality thin films, multilayers and heterostructures of a variety of multi-element material systems with rather simple technical means. We appreciate that the use of lasers to deposit films via ablation (now termed PLD) has been known since the 1960s after the invention of the first ruby laser. However, in the first two decades, PLD was something of a 'sleeping beauty' with only a few publications per year, as shown below. This state of hibernation ended abruptly with the advent of high T c superconductor research when scientists needed to grow high-quality thin films of multi-component high T c oxide systems. When most of the conventional growth techniques failed, the invention of PLD by T (Venky) Venkatesan clearly demonstrated that the newly discovered high-T c superconductor, YBa2Cu3O7-δ , could be stoichiometrically deposited as a high-quality nm-thin film with PLD [1]. As a remarkable highlight of this special issue, Venkatesan gives us his very personal reminiscence on these particularly innovative years of PLD beginning in 1986 [2]. After Venky's first paper [1], the importance of this invention was realized worldwide and the number of publications on PLD increased exponentially, as shown in figure 1. Figure 1. Figure 1. Published items per year with title or topic PLD. Data from Thomson Reuters Web of Knowledge in September 2013. After publication of Venky's famous paper in 1987 [1], the story of PLD's success began with a sudden jump in the number of publications, about 25 years ago. A first PLD textbook covering its basic understanding was soon published, in 1994, by Chrisey and Hubler [3]. Within a

  16. Laser and intense pulsed light hair removal technologies

    Haedersdal, M; Beerwerth, F; Nash, J F

    2011-01-01

    Light-based hair removal (LHR) is one of the fastest growing, nonsurgical aesthetic cosmetic procedures in the United States and Europe. A variety of light sources including lasers, e.g. alexandrite laser (755 nm), pulsed diode lasers (800, 810 nm), Nd:YAG laser (1064 nm) and broad-spectrum intense...

  17. Stimulated brillouin backscatter of a short-pulse laser

    Hinkel, D.E.; Williams, E.A.; Berger, R.L.

    1994-01-01

    Stimulated Brillouin backscattering (SBBS) from a short-pulse laser, where the pulse length is short compared to the plasma length, is found to be qualitatively different than in the long pulse regime, where the pulse length is long compared to the plasma length. We find that after an initial transient of order the laser pulse length transit time, the instability reaches a steady state in the variables x' = x - V g t, t' = t, where V g is the pulse group velocity. In contrast, SBBS in a long pulse can be absolutely unstable and grows indefinitely, or until nonlinearities intervene. We find that the motion of the laser pulse induces Doppler related effects that substantially modify the backscattered spectrum at higher intensities, where the instability is strongly coupled (i.e. , has a growth rate large compared to the ion acoustic frequency)

  18. New methods of generation of ultrashort laser pulses for ranging

    Jelinkova, Helena; Hamal, Karel; Kubecek, V.; Prochazka, Ivan

    1993-01-01

    To reach the millimeter satellite laser ranging accuracy, the goal for nineties, new laser ranging techniques have to be applied. To increase the laser ranging precision, the application of the ultrashort laser pulses in connection with the new signal detection and processing techniques, is inevitable. The two wavelength laser ranging is one of the ways to measure the atmospheric dispersion to improve the existing atmospheric correction models and hence, to increase the overall system ranging accuracy to the desired value. We are presenting a review of several nonstandard techniques of ultrashort laser pulses generation, which may be utilized for laser ranging: compression of the nanosecond pulses using stimulated Brillouin and Raman backscattering; compression of the mode-locked pulses using Raman backscattering; passive mode-locking technique with nonlinear mirror; and passive mode-locking technique with the negative feedback.

  19. Development of frequency tunable Ti:sapphire laser and dye laser pumped by a pulsed Nd:YAG laser

    Yi, Jong Hoon; Horn, Roland; Wendt, K.

    2001-01-01

    We investigated lasing characteristics of two kinds of tunable laser, liquid dye laser and solid Ti:sapphire crystal laser, pumped by high pulse repetition rate Nd:YAG laser. Dye laser showed drastically reduced pulsewidth compared with that of pump laser and it also contained large amount of amplified spontaneous emission. Ti:sapphire laser showed also reduced pulsewidth. But, the laser conversion pump laser and Ti:sapphire laser pulse, we used a Brewster-cut Pockel's cell for Q-switching. The laser was frequency doubled by a type I BBO crystal outside of the cavity.

  20. Pulse laser ablation at water-air interface

    Utsunomiya, Yuji; Kajiwara, Takashi; Nishiyama, Takashi; Nagayama, Kunihito; Kubota, Shiro

    2010-06-01

    We studied a new pulse laser ablation phenomenon on a liquid surface layer, which is caused by the difference between the refractive indices of the two materials involved. The present study was motivated by our previous study, which showed that laser ablation can occur at the interface between a transparent material and a gas or liquid medium when the laser pulse is focused through the transparent material. In this case, the ablation threshold fluence is reduced remarkably. In the present study, experiments were conducted in water and air in order to confirm this phenomenon for a combination of two fluid media with different refractive indices. This phenomenon was observed in detail by pulse laser shadowgraphy. A high-resolution film was used to record the phenomenon with a Nd:YAG pulse laser with 10-ns duration as a light source. The laser ablation phenomenon on the liquid surface layer caused by a focused Nd:YAG laser pulse with 1064-nm wavelength was found to be followed by the splashing of the liquid surface, inducing a liquid jet with many ligaments. The liquid jet extension velocity was around 1000 m/s in a typical case. The liquid jet decelerated drastically due to rapid atomization at the tips of the ligaments. The liquid jet phenomenon was found to depend on the pulse laser parameters such as the laser fluence on the liquid surface, laser energy, and laser beam pattern. The threshold laser fluence for the generation of a liquid jet was 20 J/cm2. By increasing the incident laser energy with a fixed laser fluence, the laser focused area increased, which eventually led to an increase in the size of the plasma column. The larger the laser energy, the larger the jet size and the longer the temporal behavior. The laser beam pattern was found to have significant effects on the liquid jet’s velocity, shape, and history.

  1. Pulsed power supplies for laser flashlamps. Final report

    Bird, W.L. Jr.; Driga, M.D.; Mayhall, D.J.T.; Brennan, M.

    1978-10-01

    A preliminary engineering design of a compensated pulse alternator for driving laser flashlamps is presented. The work performed by the Center for Electromechanics at The University of Texas at Austin also includes the optimization and revision of the prototype design for a compensated pulse alternator power supply for the NOVA laser system at Lawrence Livermore Laboratory

  2. Electron acceleration by a self-diverging intense laser pulse

    Singh, K.P.; Gupta, D.N.; Tripathi, V.K.; Gupta, V.L.

    2004-01-01

    Electron acceleration by a laser pulse having a Gaussian radial and temporal profile of intensity has been studied. The interaction region is vacuum followed by a gas. The starting point of the gas region has been chosen around the point at which the peak of the pulse interacts with the electron. The tunnel ionization of the gas causes a defocusing of the laser pulse and the electron experiences the action of a ponderomotive deceleration at the trailing part of the pulse with a lower intensity rather than an acceleration at the rising part of the laser pulse with a high intensity, and thus gains net energy. The initial density of the neutral gas atoms should be high enough to properly defocus the pulse; otherwise the electron experiences some deceleration during the trailing part of the pulse and the net energy gain is reduced. The rate of tunnel ionization increases with the increase in the laser intensity and the initial density of neutral gas atoms, and with the decreases in the laser spot size, which causes more defocusing of the laser pulse. The required initial density of neutral gas atoms decreases with the increase in the laser intensity and also with the decrease in the laser spot size

  3. Ultrashort laser pulses and electromagnetic pulse generation in air and on dielectric surfaces

    Sprangle, P.; Penano, J.R.; Hafizi, B.; Kapetanakos, C.A.

    2004-01-01

    Intense, ultrashort laser pulses propagating in the atmosphere have been observed to emit sub-THz electromagnetic pulses (EMPS). The purpose of this paper is to analyze EMP generation from the interaction of ultrashort laser pulses with air and with dielectric surfaces and to determine the efficiency of conversion of laser energy to EMP energy. In our self-consistent model the laser pulse partially ionizes the medium, forms a plasma filament, and through the ponderomotive forces associated with the laser pulse, drives plasma currents which are the source of the EMP. The propagating laser pulse evolves under the influence of diffraction, Kerr focusing, plasma defocusing, and energy depletion due to electron collisions and ionization. Collective effects and recombination processes are also included in the model. The duration of the EMP in air, at a fixed point, is found to be a few hundred femtoseconds, i.e., on the order of the laser pulse duration plus the electron collision time. For steady state laser pulse propagation the flux of EMP energy is nonradiative and axially directed. Radiative EMP energy is present only for nonsteady state or transient laser pulse propagation. The analysis also considers the generation of EMP on the surface of a dielectric on which an ultrashort laser pulse is incident. For typical laser parameters, the power and energy conversion efficiency from laser radiation to EMP radiation in both air and from dielectric surfaces is found to be extremely small, -8 . Results of full-scale, self-consistent, numerical simulations of atmospheric and dielectric surface EMP generation are presented. A recent experiment on atmospheric EMP generation is also simulated

  4. Electron acceleration by laser produced wake field: Pulse shape effect

    Malik, Hitendra K.; Kumar, Sandeep; Nishida, Yasushi

    2007-12-01

    Analytical expressions are obtained for the longitudinal field (wake field: Ex), density perturbations ( ne') and the potential ( ϕ) behind a laser pulse propagating in a plasma with the pulse duration of the electron plasma period. A feasibility study on the wake field is carried out with Gaussian-like (GL) pulse, rectangular-triangular (RT) pulse and rectangular-Gaussian (RG) pulse considering one-dimensional weakly nonlinear theory ( ne'/n0≪1), and the maximum energy gain acquired by an electron is calculated for all these three types of the laser pulse shapes. A comparative study infers that the RT pulse yields the best results: In its case maximum electron energy gain is 33.5 MeV for a 30 fs pulse duration whereas in case of GL (RG) pulse of the same duration the gain is 28.6 (28.8)MeV at the laser frequency of 1.6 PHz and the intensity of 3.0 × 10 18 W/m 2. The field of the wake and hence the energy gain get enhanced for the higher laser frequency, larger pulse duration and higher laser intensity for all types of the pulses.

  5. Active manipulation of the selective alignment by two laser pulses

    Zeng-Qiang, Yang; Zhi-Rong, Guo; Gui-Xian, Ge

    2010-01-01

    This paper solves numerically the full time-dependent Schrödinger equation based on the rigid rotor model, and proposes a novel strategy to determine the optimal time delay of the two laser pulses to manipulate the molecular selective alignment. The results illustrate that the molecular alignment generated by the first pulse can be suppressed or enhanced selectively, the relative populations of even and odd rotational states in the final rotational wave packet can be manipulated selectively by precisely inserting the peak of the second laser pulse at the time when the slope for the alignment parameter by the first laser locates a local maximum for the even rotational states and a local minimum for the odds, and vice versa. The selective alignment can be further optimised by selecting the intensity ratio of the two laser pulses on the condition that the total laser intensity and pulse duration are kept constant. (atomic and molecular physics)

  6. Ultrafast pulse lasers jump to macro applications

    Griebel, Martin; Lutze, Walter; Scheller, Torsten

    2016-03-01

    Ultrafast Lasers have been proven for several micro applications, e.g. stent cutting, for many years. Within its development of applications Jenoptik has started to use ultrafast lasers in macro applications in the automotive industry. The JenLas D2.fs-lasers with power output control via AOM is an ideal tool for closed loop controlled material processing. Jenoptik enhanced his well established sensor controlled laser weakening process for airbag covers to a new level. The patented process enables new materials using this kind of technology. One of the most sensitive cover materials is genuine leather. As a natural product it is extremely inhomogeneous and sensitive for any type of thermal load. The combination of femtosecond pulse ablation and closed loop control by multiple sensor array opens the door to a new quality level of defined weakening. Due to the fact, that the beam is directed by scanning equipment the process can be split in multiple cycles additionally reducing the local energy input. The development used the 5W model as well as the latest 10W release of JenLas D2.fs and achieved amazing processing speeds which directly fulfilled the requirements of the automotive industry. Having in mind that the average cycle time of automotive processes is about 60s, trials had been done of processing weakening lines in genuine leather of 1.2mm thickness. Parameters had been about 15 cycles with 300mm/s respectively resulting in an average speed of 20mm/s and a cycle time even below 60s. First samples had already given into functional and aging tests and passed successfully.

  7. Hose-Modulation Instability of Laser Pulses in Plasmas

    Sprangle, P.; Krall, J.; Esarey, E.

    1994-01-01

    A laser pulse propagating in a uniform plasma or a preformed plasma density channel is found to undergo a combination of hose and modulation instabilities, provided the pulse centroid has an initial tilt. Coupled equations for the laser centroid and envelope are derived and solved for a finite-length laser pulse. Significant coupling between the centroid and the envelope, harmonic generation in the envelope, and strong modification of the wake field can occur. Methods to reduce the growth rate of the laser hose instability are demonstrated

  8. Nonlinear laser pulse response in a crystalline lens.

    Sharma, R P; Gupta, Pradeep Kumar; Singh, Ram Kishor; Strickland, D

    2016-04-01

    The propagation characteristics of a spatial Gaussian laser pulse have been studied inside a gradient-index structured crystalline lens with constant-density plasma generated by the laser-tissue interaction. The propagation of the laser pulse is affected by the nonlinearities introduced by the generated plasma inside the crystalline lens. Owing to the movement of plasma species from a higher- to a lower-temperature region, an increase in the refractive index occurs that causes the focusing of the laser pulse. In this study, extended paraxial approximation has been applied to take into account the evolution of the radial profile of the Gaussian laser pulse. To examine the propagation characteristics, variation of the beam width parameter has been observed as a function of the laser power and initial beam radius. The cavitation bubble formation, which plays an important role in the restoration of the elasticity of the crystalline lens, has been investigated.

  9. Evaluation of cytogenetic effects of very short laser pulsed radiations

    Guedeney, G.; Courant, D.; Malarbet, J.-L.; Dolloy, M.-T.; Court, L.

    1992-01-01

    The aim of this study is to evaluate the capacity of a laser, delivering very short pulses in the near infrared spectrum with a high pulse ratio frequency, to induce genetic modification on biological tissues. Chromatid exchanges and chromosomal aberrations studies are used to test potential effect on human lymphocytes. The laser irradiation induces a significant increase of acentric fragments but the absence of dicentric suggests that a repetitive very short pulses irradiation has a relatively low capacity to induce genetic abnormalities. (author)

  10. Thermal Effect of Pulsed Laser on Human Skin

    N. C. Majumdar; V. K. Kochhar

    1985-01-01

    An attempt has been made to derive from theoretical considerations, some idea about safety limits of exposure with regard to radiant energy skin burns. This may be regarded as a preliminary enquiry in respect of thermal tissue damage by pulsed laser radiation, since the effects of isolated single pulses from ruby laser only have been considered. The study needs to be extended to other wavelengths as well as to trains of pulses.

  11. Chirp of monolithic colliding pulse mode-locked diode lasers

    Hofmann, M.; Bischoff, S.; Franck, Thorkild

    1997-01-01

    Spectrally resolved streak camera measurements of picosecond pulses emitted by hybridly colliding pulse mode-locked (CPM) laser diodes are presented in this letter. Depending on the modulation frequency both blue-chirped (upchirped) and red-chirped (downchirped) pulses can be observed. The two...... different regimes and the transition between them are characterized experimentally and the behavior is explained on the basis of our model for the CPM laser dynamics. (C) 1997 American Institute of Physics....

  12. Parametric study on femtosecond laser pulse ablation of Au films

    Ni Xiaochang; Wang Chingyue; Yang Li; Li Jianping; Chai Lu; Jia Wei; Zhang Ruobing; Zhang Zhigang

    2006-01-01

    Ablation process of 1 kHz rate femtosecond lasers (pulse duration 148 fs, wavelength 775 nm) with Au films on silica substrates has been systemically studied. The single-pulse threshold can be obtained directly. For the multiple pulses the ablation threshold varies with the number of pulses applied to the surface due to the incubation effect. From the plot of accumulated laser fluence N x φ th (N) and the number of laser pulses N, incubation coefficient of Au film can be obtained (s = 0.765). As the pulse energy is increased, the single pulse ablation rate is increasing following two ablation logarithmic regimes, which can be explained by previous research

  13. Water spray assisted ultrashort laser pulse ablation

    Silvennoinen, M.; Kaakkunen, J.J.J.; Paivasaari, K.; Vahimaa, P.

    2013-01-01

    Highlights: ► We show the novel method to use multibeam processing with ultrashort pulses efficiently. ► Sprayed thin water layer on ablation zone enhances ablation rate and quality. ► In some cases this method also enables ablation of the deeper and straighter holes compared to ones made without the water layer. ► Method also makes possible to directly write features without the self-organizing structures. - Abstract: We have studied femtosecond ablation under sprayed thin water film and its influence and benefits compared with ablation in the air atmosphere. These have been studied in case of the hole and the groove ablation using IR femtosecond laser. Water enhances the ablation rate and in some situations it makes possible to ablate the holes with a higher aspect ratio. While ablating the grooves, the water spray allows using the high fluences without the generation of the self-organized structures.

  14. Pulsed laser-induced SEU in integrated circuits

    Buchner, S.; Kang, K.; Stapor, W.J.; Campbell, A.B.; Knudson, A.R.; McDonald, P.; Rivet, S.

    1990-01-01

    The authors have used a pulsed picosecond laser to measure the threshold for single event upset (SEU) and single event latchup (SEL) for two different kinds of integrated circuits. The relative thresholds show good agreement with published ion upset data. The consistency of the results together with the advantages of using a laser system suggest that the pulsed laser can be used for SEU/SEL hardness assurance of integrated circuits

  15. Characteristics and Applications of Spatiotemporally Focused Femtosecond Laser Pulses

    Chenrui Jing

    2016-12-01

    Full Text Available Simultaneous spatial and temporal focusing (SSTF of femtosecond laser pulses gives rise to strong suppression of nonlinear self-focusing during the propagation of the femtosecond laser beam. In this paper, we begin with an introduction of the principle of SSTF, followed by a review of our recent experimental results on the characterization and application of the spatiotemporally focused pulses for femtosecond laser micromachining. Finally, we summarize all of the results and give a future perspective of this technique.

  16. CO2 laser pulse shortening by laser ablation of a metal target

    Donnelly, T.; Mazoyer, M.; Lynch, A.; O'Sullivan, G.; O'Reilly, F.; Dunne, P.; Cummins, T.

    2012-01-01

    A repeatable and flexible technique for pulse shortening of laser pulses has been applied to transversely excited atmospheric (TEA) CO 2 laser pulses. The technique involves focusing the laser output onto a highly reflective metal target so that plasma is formed, which then operates as a shutter due to strong laser absorption and scattering. Precise control of the focused laser intensity allows for timing of the shutter so that different temporal portions of the pulse can be reflected from the target surface before plasma formation occurs. This type of shutter enables one to reduce the pulse duration down to ∼2 ns and to remove the low power, long duration tails that are present in TEA CO 2 pulses. The transmitted energy is reduced as the pulse duration is decreased but the reflected power is ∼10 MW for all pulse durations. A simple laser heating model verifies that the pulse shortening depends directly on the plasma formation time, which in turn is dependent on the applied laser intensity. It is envisaged that this plasma shutter will be used as a tool for pulse shaping in the search for laser pulse conditions to optimize conversion efficiency from laser energy to useable extreme ultraviolet (EUV) radiation for EUV source development.

  17. Concave pulse shaping of a circularly polarized laser pulse from non-uniform overdense plasmas

    Hur, Min Sup [School of Natural Science, UNIST, BanYeon-Ri 100, Ulju-gun, Ulsan, 689-798 (Korea, Republic of); Kulagin, Victor V. [Sternberg Astronomical Institute, Moscow State University, Universitetsky prosp. 13, Moscow, 119992 (Russian Federation); Suk, Hyyong, E-mail: hysuk@gist.ac.kr [Department of Physics and Photon Science, GIST, 123 Cheomdan-gwangiro, Buk-gu, Gwangju, 500-712 (Korea, Republic of)

    2015-03-20

    Pulse shaping of circularly polarized laser pulses in nonuniform overdense plasmas are investigated numerically. Specifically we show by two-dimensional particle-in-cell simulations the generation of a concave pulse front of a circularly polarized, a few tens of petawatt laser pulse from a density-tapered, overdense plasma slab. The concept used for the transverse-directional shaping is the differential transmittance depending on the plasma density, and the laser intensity. For suitable selection of the slab parameters for the concave pulse shaping, we studied numerically the pulse transmittance, which can be used for further parameter design of the pulse shaping. The concavely shaped circularly polarized pulse is expected to add more freedom in controlling the ion-beam characteristics in the RPDA regime. - Highlights: • Laser pulse shaping for a concave front by non-uniform overdense plasma was studied. • Particle-in-cell (PIC) simulations were used for the investigation. • A laser pulse can be shaped by a density-tapered overdense plasma. • The concave and sharp pulse front are useful in many laser–plasma applications. • They are important for ion acceleration, especially in the radiation pressure dominant regime.

  18. Control of laser pulse waveform in longitudinally excited CO2 laser by adjustment of excitation circuit

    Uno, Kazuyuki; Jitsuno, Takahisa

    2018-05-01

    In a longitudinally excited CO2 laser that had a 45 cm-long discharge tube with a 1:1:2 mixture of CO2/N2/He gas at a pressure of 3.0 kPa, we realized the generation of a short laser pulse with a spike pulse width of about 200 ns and a pulse tail length of several tens of microseconds, control of the energy ratio of the spike pulse part to the pulse tail part in the short laser pulse, the generation of a long laser pulse with a pulse width of several tens of microseconds, and control of the pulse width in the long laser pulse, by using four types of excitation circuits in which the capacitance was adjusted. In the short laser pulse, the energy ratio was in the range 1:14-1:112. In the long laser pulse, the pulse width was in the range 25.7-82.7 μs.

  19. CTS and CZTS for solar cells made by pulsed laser deposition and pulsed electron deposition

    Ettlinger, Rebecca Bolt

    This thesis concerns the deposition of thin films for solar cells using pulsed laser deposition (PLD) and pulsed electron deposition (PED). The aim was to deposit copper tin sulfide (CTS) and zinc sulfide (ZnS) by pulsed laser deposition to learn about these materials in relation to copper zinc tin...... time. We compared the results of CZTS deposition by PLD at DTU in Denmark to CZTS made by PED at IMEM-CNR, where CIGS solar cells have successfully been fabricated at very low processing temperatures. The main results of this work were as follows: Monoclinic-phase CTS films were made by pulsed laser...... deposition followed by high temperature annealing. The films were used to understand the double band gap that we and other groups observed in the material. The Cu-content of the CTS films varied depending on the laser fluence (the laser energy per pulse and per area). The material transfer from...

  20. Pulsed laser dewetting of nickel catalyst for carbon nanofiber growth

    Guan, Y F; Pearce, R C; Simpson, M L; Rack, P D; Melechko, A V; Hensley, D K

    2008-01-01

    We present a pulsed laser dewetting technique that produces single nickel catalyst particles from lithographically patterned disks for subsequent carbon nanofiber growth through plasma enhanced chemical vapor deposition. Unlike the case for standard heat treated Ni catalyst disks, for which multiple nickel particles and consequently multiple carbon nanofibers (CNFs) are observed, single vertically aligned CNFs could be obtained from the laser dewetted catalyst. Different laser dewetting parameters were tested in this study, such as the laser energy density and the laser processing time measured by the total number of laser pulses. Various nickel disk radii and thicknesses were attempted and the resultant number of carbon nanofibers was found to be a function of the initial disk dimension and the number of laser pulses

  1. New laser system for highly sensitive clinical pulse oximetry

    Hamza, Mostafa; Hamza, Mohammad

    1996-04-01

    This paper describes the theory and design of a new pulse oximeter in which laser diodes and other compact laser sources are used for the measurement of oxygen saturation in patients who are at risk of developing hypoxemia. The technique depends upon illuminating special sites of the skin of the patient with radiation from modulated laser sources at selected wavelengths. The specific laser wavelengths are chosen based on the absorption characteristics of oxyhemoglobin, reduced hemoglobin and other interfering sources for obtaining more accurate measurements. The laser radiation transmitted through the tissue is detected and signal processing based on differential absorption laser spectroscopy is done in such a way to overcome the primary performance limitations of the conventionally used pulse oximetry. The new laser pulse oximeter can detect weak signals and is not affected by other light sources such as surgical lamps, phototherapy units, etc. The detailed description and operating characteristics of this system are presented.

  2. Energy losses estimation during pulsed-laser seam welding

    Šebestová, Hana; Havelková, M.; Chmelíčková, H.

    2014-01-01

    Roč. 45, č. 3 (2014), s. 1116-1121 ISSN 1073-5615 R&D Projects: GA MŠk(CZ) LG13007 Institutional support: RVO:68378271 Keywords : laser welding * pulsed-laser * Nd:YAG laser Subject RIV: JP - Industrial Processing Impact factor: 1.461, year: 2014

  3. Optical pulse generation using fiber lasers and integrated optics

    Wilcox, R.B.; Browning, D.F.; Burkhart, S.C.; VanWonterghem, B.W.

    1995-01-01

    We have demonstrated an optical pulse forming system using fiber and integrated optics, and have designed a multiple-output system for a proposed fusion laser facility. Our approach is an advancement over previous designs for fusion lasers, and an unusual application of fiber lasers and integrated optics

  4. Explosive Nucleosynthesis Study Using Laser Driven γ-ray Pulses

    Takehito Hayakawa

    2017-03-01

    Full Text Available We propose nuclear experiments using γ-ray pulses provided from high field plasma generated by high peak power laser. These γ-ray pulses have the excellent features of extremely short pulse, high intensity, and continuous energy distribution. These features are suitable for the study of explosive nucleosyntheses in novae and supernovae, such as the γ process and ν process. We discuss how to generate suitable γ-ray pulses and the nuclear astrophysics involved.

  5. Fragmentation dynamics of molecular hydrogen in strong ultrashort laser pulses

    Rudenko, A; Feuerstein, B; Zrost, K; Jesus, V L B de; Ergler, T; Dimopoulou, C; Schroeter, C D; Moshammer, R; Ullrich, J

    2005-01-01

    We present the results of a systematic experimental study of dissociation and Coulomb explosion of molecular hydrogen induced by intense ultrashort (7-25 fs) laser pulses. Using coincident recoil-ion momentum spectroscopy we can distinguish the contributions from dissociation and double ionization even if they result in the same kinetic energies of the fragments. The dynamics of all fragmentation channels drastically depends on the pulse duration, and for 7 fs pulses becomes extremely sensitive to the pulse shape

  6. Laser cleaning of pulsed laser deposited rhodium films for fusion diagnostic mirrors

    Uccello, A.; Maffini, A.; Dellasega, D.; Passoni, M.

    2013-01-01

    Highlights: ► Pulsed laser deposition is exploited to produce Rh films for first mirrors. ► Pulsed laser deposition is exploited to produce tokamak-like C contaminants. ► Rh laser damage threshold has been evaluated for infrared pulses. ► Laser cleaning of C contaminated Rh films gives promising results. -- Abstract: In this paper an experimental investigation on the laser cleaning process of rhodium films, potentially candidates to be used as tokamak first mirrors (FMs), from redeposited carbon contaminants is presented. A relevant issue that lowers mirror's performance during tokamak operations is the redeposition of sputtered material from the first wall on their surface. Among all the possible techniques, laser cleaning, in which a train of laser pulses is launched to the surface that has to be treated, is a method to potentially mitigate this problem. The same laser system (Q-switched Nd:YAG laser with a fundamental wavelength of 1064-nm and 7-ns pulses) has been employed with three aims: (i) production by pulsed laser deposition (PLD) of Rh film mirrors, (ii) production by PLD of C deposits with controlled morphology, and (iii) investigation of the laser cleaning method onto C contaminated Rh samples. The evaluation of Rh films laser damage threshold, as a function of fluence and number of pulses, is discussed. Then, the C/Rh films have been cleaned by the laser beam. The exposed zones have been characterized by visual inspection and scanning electron microscopy (SEM), showing promising results

  7. Effect of laser pulse parameters on the size and fluorescence of nanodiamonds formed upon pulsed-laser irradiation

    Bai, Peikang [School of Materials Science and Engineering, North University of China, Taiyuan 030051 (China); Hu, Shengliang, E-mail: hsliang@yeah.net [Key Laboratory of Instrumentation Science and Dynamic Measurement (North University of China), Ministry of Education, National Key Laboratory Science and Technology on Electronic Test and Measurement, Taiyuan 030051 (China); School of Materials Science and Engineering, North University of China, Taiyuan 030051 (China); Zhang, Taiping; Sun, Jing [School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China); Cao, Shirui [School of Materials Science and Engineering, North University of China, Taiyuan 030051 (China)

    2010-07-15

    The size of nanodiamonds formed upon laser irradiation could be easily controlled over simply adjusting laser pulse parameters. The stable size and structure of nanodiamonds were mostly determined by laser power density and pulse width. Both large nanodiamonds with multiply twinning structure (MTS) and small nanodiamonds with single crystalline structure (SCS) emitted strong visible light after surface passivation, and their fluorescence quantum yield (QY) was 4.6% and 7.1%, respectively.

  8. Effect of laser pulse parameters on the size and fluorescence of nanodiamonds formed upon pulsed-laser irradiation

    Bai, Peikang; Hu, Shengliang; Zhang, Taiping; Sun, Jing; Cao, Shirui

    2010-01-01

    The size of nanodiamonds formed upon laser irradiation could be easily controlled over simply adjusting laser pulse parameters. The stable size and structure of nanodiamonds were mostly determined by laser power density and pulse width. Both large nanodiamonds with multiply twinning structure (MTS) and small nanodiamonds with single crystalline structure (SCS) emitted strong visible light after surface passivation, and their fluorescence quantum yield (QY) was 4.6% and 7.1%, respectively.

  9. Pulse generation and preamplification for long pulse beamlines of Orion laser facility.

    Hillier, David I; Winter, David N; Hopps, Nicholas W

    2010-06-01

    We describe the pulse generation, shaping, and preamplification system for the nanosecond beamlines of the Orion laser facility. The system generates shaped laser pulses of up to approximately 1 J of 100 ps-5 ns duration with a programmable temporal profile. The laser has a 30th-power supergaussian spatial profile and is diffraction limited. The system is capable of imposing 2D smoothing by spectral dispersion upon the beam, which will produce a nonuniformity of 10% rms at the target.

  10. Generation and amplification of nanaosecond duration multiline hf laser pulses

    Getzinger, R.L.; Ware, K.D.; Carpenter, J.P.

    1976-01-01

    High-power, fast-rising pulses of hydrogen fluoride laser energy suitable for laser-fusion target interaction experiments can in principle be generated by directing an electro-optically shuttered oscillator pulse through one or more electron-beam driven amplifiers. A three-stage HF master oscillator-power amplifier (MOPA) configuration was constructed and tested using SF 6 -C 2 H 6 in which an E-O generated 4-ns-FWHM pulse was amplified in an electron-beam-excited third stage and subsequently isolated with a Brewster angle splitter. Independent experiments in which a 100-ns-FWHM pilot pulse interacted with the power amplifier demonstrated for the first time complete extraction of the available laser energy. These two results provide strong evidence that with upgrading to H 2 -F 2 , it should be possible to obtain nanosecond duration pulses with power levels sufficient for meaningful laser fusion target coupling experiments

  11. Short-pulse laser interactions with disordered materials and liquids

    Phinney, L.M.; Goldman, C.H.; Longtin, J.P.; Tien, C.L. [Univ. of California, Berkeley, CA (United States)

    1995-12-31

    High-power, short-pulse lasers in the picosecond and subpicosecond range are utilized in an increasing number of technologies, including materials processing and diagnostics, micro-electronics and devices, and medicine. In these applications, the short-pulse radiation interacts with a wide range of media encompassing disordered materials and liquids. Examples of disordered materials include porous media, polymers, organic tissues, and amorphous forms of silicon, silicon nitride, and silicon dioxide. In order to accurately model, efficiently control, and optimize short-pulse, laser-material interactions, a thorough understanding of the energy transport mechanisms is necessary. Thus, fractals and percolation theory are used to analyze the anomalous diffusion regime in random media. In liquids, the thermal aspects of saturable and multiphoton absorption are examined. Finally, a novel application of short-pulse laser radiation to reduce surface adhesion forces in microstructures through short-pulse laser-induced water desorption is presented.

  12. Prepulse effect on intense femtosecond laser pulse propagation in gas

    Giulietti, Antonio; Tomassini, Paolo; Galimberti, Marco; Giulietti, Danilo; Gizzi, Leonida A.; Koester, Petra; Labate, Luca; Ceccotti, Tiberio; D'Oliveira, Pascal; Auguste, Thierry; Monot, Pascal; Martin, Philippe

    2006-01-01

    The propagation of an ultrashort laser pulse can be affected by the light reaching the medium before the pulse. This can cause a serious drawback to possible applications. The propagation in He of an intense 60-fs pulse delivered by a Ti:sapphire laser in the chirped pulse amplification (CPA) mode has been investigated in conditions of interest for laser-plasma acceleration of electrons. The effects of both nanosecond amplified spontaneous emission and picosecond pedestals have been clearly identified. There is evidence that such effects are basically of refractive nature and that they are not detrimental for the propagation of a CPA pulse focused to moderately relativistic intensity. The observations are fully consistent with numerical simulations and can contribute to the search of a stable regime for laser acceleration

  13. Research on temperature characteristics of laser energy meter absorber irradiated by ms magnitude long pulse laser

    Li, Nan; Qiao, Chunhong; Fan, Chengyu; Zhang, Jinghui; Yang, Gaochao

    2017-10-01

    The research on temperature characteristics for large-energy laser energy meter absorber is about continuous wave (CW) laser before. For the measuring requirements of millisecond magnitude long pulse laser energy, the temperature characteristics for absorber are numerically calculated and analyzed. In calculation, the temperature field distributions are described by heat conduction equations, and the metal cylinder cavity is used for absorber model. The results show that, the temperature of absorber inwall appears periodic oscillation with pulse structure, the oscillation period and amplitude respectively relate to the pulse repetition frequency and single pulse energy. With the wall deep increasing, the oscillation amplitude decreases rapidly. The temperature of absorber outerwall is without periodism, and rises gradually with time. The factors to affect the temperature rise of absorber are single pulse energy, pulse width and repetition frequency. When the laser irradiation stops, the temperature between absorber inwall and outerwall will reach agreement rapidly. After special technology processing to enhance the capacity of resisting laser damage for absorber inwall, the ms magnitude long pulse laser energy can be obtained with the method of measuring the temperature of absorber outerwall. Meanwhile, by optimization design of absorber structure, when the repetition frequency of ms magnitude pulse laser is less than 10Hz, the energy of every pulse for low repetition frequency pulse sequence can be measured. The work offers valuable references for the design of ms magnitude large-energy pulse laser energy meter.

  14. Efficient delivery of 60 J pulse energy of long pulse Nd:YAG laser ...

    2014-02-09

    Feb 9, 2014 ... Most of today's industrial Nd:YAG lasers use fibre-optic beam delivery. ... optical fibre and successfully delivered up to 60 J of pulse energy with .... and electrical pump input to laser output conversion efficiency is about 5%. ... [3] W Koechner, Solid state laser engineering, 5th edn (Springer, Berlin, 1999).

  15. Double pulse laser ablation and plasma: Laser induced breakdown spectroscopy signal enhancement

    Babushok, V.I.; DeLucia, F.C.; Gottfried, J.L.; Munson, C.A.; Miziolek, A.W.

    2006-01-01

    A review of recent results of the studies of double laser pulse plasma and ablation for laser induced breakdown spectroscopy applications is presented. The double pulse laser induced breakdown spectroscopy configuration was suggested with the aim of overcoming the sensitivity shortcomings of the conventional single pulse laser induced breakdown spectroscopy technique. Several configurations have been suggested for the realization of the double pulse laser induced breakdown spectroscopy technique: collinear, orthogonal pre-spark, orthogonal pre-heating and dual pulse crossed beam modes. In addition, combinations of laser pulses with different wavelengths, different energies and durations were studied, thus providing flexibility in the choice of wavelength, pulse width, energy and pulse sequence. The double pulse laser induced breakdown spectroscopy approach provides a significant enhancement in the intensity of laser induced breakdown spectroscopy emission lines up to two orders of magnitude greater than a conventional single pulse laser induced breakdown spectroscopy. The double pulse technique leads to a better coupling of the laser beam with the plasma plume and target material, thus providing a more temporally effective energy delivery to the plasma and target. The experimental results demonstrate that the maximum effect is obtained at some optimum separation delay time between pulses. The optimum value of the interpulse delay depends on several factors, such as the target material, the energy level of excited states responsible for the emission, and the type of enhancement process considered. Depending on the specified parameter, the enhancement effects were observed on different time scales ranging from the picosecond time level (e.g., ion yield, ablation mass) up to the hundred microsecond level (e.g., increased emission intensity for laser induced breakdown spectroscopy of submerged metal target in water). Several suggestions have been proposed to explain

  16. Higher order harmonic generation in the intense laser pulse

    Parvizi, R.; Bahrampour, A.; Karimi, M.

    2006-01-01

    The high intensity pulse of laser field ionizes the atoms and electrons are going to the continuum states of atoms. electrons absorb energy from the strong laser field. The back ground electromagnetic field causes to come back the electrons to ground states of atoms and the absorbed energy is emitted as a high order odd harmonics of incident light. The intensity of emitted harmonics depends on the material atoms and the laser pulse shape. I this paper the effects of step pulse duration on the high order harmonic radiated by the Argon, Helium, and Hydrogen atoms are reported.

  17. Development of high power pulsed CO2 laser

    Nakai, Sadao; Matoba, Masafumi; Fujita, Hisanori; Daido, Hiroyuki; Inoue, Mitsuo

    1982-01-01

    The inertial nuclear fusion research using pellet implosion has rapidly progressed accompanying laser technique improvement and output increase. As the high output lasers for this purpose, Nd glass lasers or CO 2 lasers are used. The CO 2 lasers possess the characteristics required as reactor lasers, i.e., high efficiency, high frequency repetition, possibility of scale-up and economy. So, the technical development of high power CO 2 lasers assuming also as reactor drivers has been performed at a quick pace together with the research on the improvement of efficiency of pellet implosion by 10 μm laser beam. The Institute of Laser Engineering, Osaka University, stated to build a laser system LEKKO No. 8 of 8 beams and 10 kJ based on the experiences in laser systems LEKKO No. 1 and LEKKO No. 2, and the system LEKKO No. 8 was completed in March, 1981. The operation tests for one year since then has indicated as the laser characteristics that the system performance was as designed initially. This paper reviews the structure, problems and present status of the large scale CO 2 lasers. In other words, the construction of laser system, CO 2 laser proper, oscillator, booster amplifier, prevention of parasitic oscillation, non-linear pulse propagation and fairing of output pulse form, system control and beam alignment, and high power problems are described. The results obtained are to be reported in subsequent issues. (Wakatsuki, Y.)

  18. Stabilizing laser energy density on a target during pulsed laser deposition of thin films

    Dowden, Paul C.; Jia, Quanxi

    2016-05-31

    A process for stabilizing laser energy density on a target surface during pulsed laser deposition of thin films controls the focused laser spot on the target. The process involves imaging an image-aperture positioned in the beamline. This eliminates changes in the beam dimensions of the laser. A continuously variable attenuator located in between the output of the laser and the imaged image-aperture adjusts the energy to a desired level by running the laser in a "constant voltage" mode. The process provides reproducibility and controllability for deposition of electronic thin films by pulsed laser deposition.

  19. Pulsed-laser atom-probe field-ion microscopy

    Kellogg, G.L.; Tsong, T.T.

    1980-01-01

    A time-of-flight atom-probe field-ion microscope has been developed which uses nanosecond laser pulses to field evaporate surface species. The ability to operate an atom-probe without using high-voltage pulses is advantageous for several reasons. The spread in energy arising from the desorption of surface species prior to the voltage pulse attaining its maximum amplitude is eliminated, resulting in increased mass resolution. Semiconductor and insulator samples, for which the electrical resistivity is too high to transmit a short-duration voltage pulse, can be examined using pulsed-laser assisted field desorption. Since the electric field at the surface can be significantly smaller, the dissociation of molecular adsorbates by the field can be reduced or eliminated, permitting well-defined studies of surface chemical reactions. In addition to atom-probe operation, pulsed-laser heating of field emitters can be used to study surface diffusion of adatoms and vacancies over a wide range of temperatures. Examples demonstrating each of these advantages are presented, including the first pulsed-laser atom-probe (PLAP) mass spectra for both metals (W, Mo, Rh) and semiconductors (Si). Molecular hydrogen, which desorbs exclusively as atomic hydrogen in the conventional atom probe, is shown to desorb undissociatively in the PLAP. Field-ion microscope observations of the diffusion and dissociation of atomic clusters, the migration of adatoms, and the formation of vacancies resulting from heating with a 7-ns laser pulse are also presented

  20. Photodisruption in biological tissues using femtosecond laser pulses

    Shen, Nan

    Transparent materials do not ordinarily absorb visible or near-infrared light. However, the intensity of a tightly focused femtosecond laser pulse is great enough that nonlinear absorption of the laser energy takes place in transparent materials, leading to optical breakdown and permanent material modification. Because the absorption process is nonlinear, absorption and material modification are confined to the extremely small focal volume. Optical breakdown in transparent or semi-transparent biological tissues depends on intensity rather than energy. As a result, focused femtosecond pulses induce optical breakdown with significantly less pulse energy than is required with longer pulses. The use of femtosecond pulses therefore minimizes the amount of energy deposited into the targeted region of the sample, minimizing mechanical and thermal effects that lead to collateral damage in adjacent tissues. We demonstrate photodisruptive surgery in animal skin tissue and single cells using 100-fs laser pulses. In mouse skin, we create surface incisions and subsurface cavities with much less collateral damage to the surrounding tissue than is produced with picosecond pulses. Using pulses with only a few nanojoules of energy obtained from an unamplified femtosecond oscillator, we destroy single mitochondria in live cells without affecting cell viability, providing insights into the structure of the mitochondrial network. An apparatus is constructed to perform subcellular surgery and multiphoton 3D laser scanning imaging simultaneously with a single laser and objective lens.