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Sample records for force correlation spectroscopy

  1. Acoustic force spectroscopy

    Sitters, G.; Kamsma, D.; Thalhammer, G.; Ritsch-Marte, M.; Peterman, E.J.G.; Wuite, G.J.L.

    2015-01-01

    Force spectroscopy has become an indispensable tool to unravel the structural and mechanochemical properties of biomolecules. Here we extend the force spectroscopy toolbox with an acoustic manipulation device that can exert forces from subpiconewtons to hundreds of piconewtons on thousands of

  2. Local mobility in lipid domains of supported bilayers characterized by atomic force microscopy and fluorescence correlation spectroscopy.

    Frankel, Daniel J.; Buranda, T. (University of New Mexico, Albuquerque, NM); Burns, Alan Richard

    2005-01-01

    Fluorescence correlation spectroscopy (FCS) is used to examine mobility of labeled probes at specific sites in supported bilayers consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid domains in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Those sites are mapped beforehand with simultaneous atomic force microscopy and submicron confocal fluorescence imaging, allowing characterization of probe partitioning between gel DPPC and disordered liquid DOPC domains with corresponding topography of domain structure. We thus examine the relative partitioning and mobility in gel and disordered liquid phases for headgroup- and tailgroup-labeled GM1 ganglioside probes and for headgroup- and tailgroup-labeled phospholipid probes. For the GM1 probes, large differences in mobility between fluid and gel domains are observed; whereas unexpected mobility is observed in submicron gel domains for the phospholipid probes. We attribute the latter to domain heterogeneities that could be induced by the probe. Furthermore, fits to the FCS data for the phospholipid probes in the DOPC fluid phase require two components (fast and slow). Although proximity to the glass substrate may be a factor, local distortion of the probe by the fluorophore could also be important. Overall, we observe nonideal aspects of phospholipid probe mobility and partitioning that may not be restricted to supported bilayers.

  3. Handbook of Molecular Force Spectroscopy

    Noy, Aleksandr

    2008-01-01

    "...Noy's Handbook of Molecular Force Spectroscopy is both a timely and useful summary of fundamental aspects of molecular force spectroscopy, and I believe it would make a worthwhile addition to any good scientific library. New research groups that are entering this field would be well advisedto study this handbook in detail before venturing into the exciting and challenging world of molecular force spectroscopy." Matthew F. Paige, University of Saskatchewan, Journal of the American Chemical Society Modern materials science and biophysics are increasingly focused on studying and controlling intermolecular interactions on the single-molecule level. Molecular force spectroscopy was developed in the past decade as the result of several unprecedented advances in the capabilities of modern scientific instrumentation, and defines a number of techniques that use mechanical force measurements to study interactions between single molecules and molecular assemblies in chemical and biological systems. Examples of these...

  4. Packing force data correlations

    Heiman, S.M.

    1994-01-01

    One of the issues facing valve maintenance personnel today deals with an appropriate methodology for installing and setting valve packing that will minimize leak rates, yet ensure functionality of the the valve under all anticipated operating conditions. Several variables can affect a valve packing's ability to seal, such as packing bolt torque, stem finish, and lubrication. Stem frictional force can be an excellent overall indicator of some of the underlying conditions that affect the sealing characteristics of the packing and the best parameter to use when adjusting the packing. This paper addresses stem friction forces, analytically derives the equations related to these forces, presents a methodology for measuring these forces on valve stems, and attempts to correlate the data directly to the underlying variables

  5. Variable angle correlation spectroscopy

    Lee, Y.K.; Lawrence Berkeley Lab., CA

    1994-05-01

    In this dissertation, a novel nuclear magnetic resonance (NMR) technique, variable angle correlation spectroscopy (VACSY) is described and demonstrated with 13 C nuclei in rapidly rotating samples. These experiments focus on one of the basic problems in solid state NMR: how to extract the wealth of information contained in the anisotropic component of the NMR signal while still maintaining spectral resolution. Analysis of the anisotropic spectral patterns from poly-crystalline systems reveal information concerning molecular structure and dynamics, yet in all but the simplest of systems, the overlap of spectral patterns from chemically distinct sites renders the spectral analysis difficult if not impossible. One solution to this problem is to perform multi-dimensional experiments where the high-resolution, isotropic spectrum in one dimension is correlated with the anisotropic spectral patterns in the other dimensions. The VACSY technique incorporates the angle between the spinner axis and the static magnetic field as an experimental parameter that may be incremented during the course of the experiment to help correlate the isotropic and anisotropic components of the spectrum. The two-dimensional version of the VACSY experiments is used to extract the chemical shift anisotropy tensor values from multi-site organic molecules, study molecular dynamics in the intermediate time regime, and to examine the ordering properties of partially oriented samples. The VACSY technique is then extended to three-dimensional experiments to study slow molecular reorientations in a multi-site polymer system

  6. Molecular Force Spectroscopy on Cells

    Liu, Baoyu; Chen, Wei; Zhu, Cheng

    2015-04-01

    Molecular force spectroscopy has become a powerful tool to study how mechanics regulates biology, especially the mechanical regulation of molecular interactions and its impact on cellular functions. This force-driven methodology has uncovered a wealth of new information of the physical chemistry of molecular bonds for various biological systems. The new concepts, qualitative and quantitative measures describing bond behavior under force, and structural bases underlying these phenomena have substantially advanced our fundamental understanding of the inner workings of biological systems from the nanoscale (molecule) to the microscale (cell), elucidated basic molecular mechanisms of a wide range of important biological processes, and provided opportunities for engineering applications. Here, we review major force spectroscopic assays, conceptual developments of mechanically regulated kinetics of molecular interactions, and their biological relevance. We also present current challenges and highlight future directions.

  7. Principles and applications of force spectroscopy using atomic force microscopy

    Kim, Young Kyu; Kim, Woong; Park, Joon Won [Dept. of Chemistry, Pohang University of Science and Technology, Pohang (Korea, Republic of)

    2016-12-15

    Single-molecule force spectroscopy is a powerful technique for addressing single molecules. Unseen structures and dynamics of molecules have been elucidated using force spectroscopy. Atomic force microscope (AFM)-based force spectroscopy studies have provided picoNewton force resolution, subnanometer spatial resolution, stiffness of substrates, elasticity of polymers, and thermodynamics and kinetics of single-molecular interactions. In addition, AFM has enabled mapping the distribution of individual molecules in situ, and the quantification of single molecules has been made possible without modification or labeling. In this review, we describe the basic principles, sample preparation, data analysis, and applications of AFM-based force spectroscopy and its future.

  8. Correlation ion mobility spectroscopy

    Pfeifer, Kent B [Los Lunas, NM; Rohde, Steven B [Corrales, NM

    2008-08-26

    Correlation ion mobility spectrometry (CIMS) uses gating modulation and correlation signal processing to improve IMS instrument performance. Closely spaced ion peaks can be resolved by adding discriminating codes to the gate and matched filtering for the received ion current signal, thereby improving sensitivity and resolution of an ion mobility spectrometer. CIMS can be used to improve the signal-to-noise ratio even for transient chemical samples. CIMS is especially advantageous for small geometry IMS drift tubes that can otherwise have poor resolution due to their small size.

  9. Force spectroscopy of hepatocytic extracellular matrix components

    Yongsunthon, R., E-mail: YongsuntR@Corning.com [Corning Incorporated, SP-FR-01, R1S32D, Corning, NY 14831 (United States); Baker, W.A.; Bryhan, M.D.; Baker, D.E.; Chang, T.; Petzold, O.N.; Walczak, W.J.; Liu, J.; Faris, R.A.; Senaratne, W.; Seeley, L.A.; Youngman, R.E. [Corning Incorporated, SP-FR-01, R1S32D, Corning, NY 14831 (United States)

    2009-07-15

    We present atomic force microscopy and force spectroscopy data of live hepatocytes (HEPG2/C3A liver cell line) grown in Eagle's Minimum Essential Medium, a complex solution of salts and amino acids commonly used for cell culture. Contact-mode imaging and force spectroscopy of this system allowed correlation of cell morphology and extracellular matrix (ECM) properties with substrate properties. Force spectroscopy analysis of cellular 'footprints' indicated that the cells secrete large polymers (e.g., 3.5 {mu}m contour length and estimated MW 1000 kDa) onto their substrate surface. Although definitive identification of the polymers has not yet been achieved, fluorescent-labeled antibody staining has specified the presence of ECM proteins such as collagen and laminin in the cellular footprints. The stretched polymers appear to be much larger than single molecules of known ECM components, such as collagen and heparan sulfate proteoglycan, thus suggesting that the cells create larger entangled, macromolecular structures from smaller components. There is strong evidence which suggests that the composition of the ECM is greatly influenced by the hydrophobicity of the substrate surface, with preferential production and/or adsorption of larger macromolecules on hydrophobic surfaces.

  10. PLA-PEG nanocapsules radiolabeled with 99mTechnetium-HMPAO: release properties and physicochemical characterization by atomic force microscopy and photon correlation spectroscopy.

    Pereira, Maira Alves; Mosqueira, Vanessa Carla Furtado; Vilela, José Mário Carneiro; Andrade, Margareth Spangler; Ramaldes, Gilson Andrade; Cardoso, Valbert Nascimento

    2008-01-01

    The present work describes the preparation, characterization and labelling of conventional and surface-modified nanocapsules (NC) with 99m Tc-HMPAO. The size, size distribution and homogeneity were determined by photon correlation spectroscopy (PCS) and zeta potential by laser doppler anemometry. The morphology and the structural organization were evaluated by atomic force microscopy (AFM). The stability and release profile of the NC were determined in vitro in plasma. The results showed that the use of methylene blue induces significant increase in the encapsulation efficiency of 99m Tc-HMPAO, from 24.4 to 49.8% in PLA NC and 22.37 to 52.93% in the case of PLA-PEG NC (P<0.05) by improving the complex stabilization. The average diameter of NC calculated by PCS varied from 216 to 323 nm, while the average diameter determined by AFM varied from 238 to 426 nm. The AFM analysis of diameter/height ratios suggested a greater homogeneity of the surface-modified PLA-PEG nanocapsules compared to PLA NC concerning their flattening properties. The in vitro release of the 99m Tc-HMPAO in plasma medium was faster for the conventional PLA NC than for the surface-modified NC. For the latter, 60% of the radioactivity remained associated with NC, even after 12h of incubation. The results suggest that the surface-modified 99m Tc-HMPAO-PLA-PEG NC was more stable against label leakage in the presence of proteins and could present better performance as radiotracer in vivo.

  11. Small amplitude atomic force spectroscopy

    de Beer, Sissi; van den Ende, Henricus T.M.; Ebeling, Daniel; Mugele, Friedrich Gunther; Bhushan, Bharat

    2011-01-01

    Over the years atomic force microscopy has developed from a pure imaging technique to a tool that can be employed for measuring quantitative tip–sample interaction forces. In this chapter we provide an overview of various techniques to extract quantitative tip–sample forces focusing on both

  12. Atomic Force Microscope for Imaging and Spectroscopy

    Pike, W. T.; Hecht, M. H.; Anderson, M. S.; Akiyama, T.; Gautsch, S.; deRooij, N. F.; Staufer, U.; Niedermann, Ph.; Howald, L.; Mueller, D.

    2000-01-01

    We have developed, built, and tested an atomic force microscope (AFM) for extraterrestrial applications incorporating a micromachined tip array to allow for probe replacement. It is part of a microscopy station originally intended for NASA's 2001 Mars lander to identify the size, distribution, and shape of Martian dust and soil particles. As well as imaging topographically down to nanometer resolution, this instrument can be used to reveal chemical information and perform infrared and Raman spectroscopy at unprecedented resolution.

  13. Single molecule atomic force microscopy and force spectroscopy of chitosan.

    Kocun, Marta; Grandbois, Michel; Cuccia, Louis A

    2011-02-01

    Atomic force microscopy (AFM) and AFM-based force spectroscopy was used to study the desorption of individual chitosan polymer chains from substrates with varying chemical composition. AFM images of chitosan adsorbed onto a flat mica substrate show elongated single strands or aggregated bundles. The aggregated state of the polymer is consistent with the high level of flexibility and mobility expected for a highly positively charged polymer strand. Conversely, the visualization of elongated strands indicated the presence of stabilizing interactions with the substrate. Surfaces with varying chemical composition (glass, self-assembled monolayer of mercaptoundecanoic acid/decanethiol and polytetrafluoroethylene (PTFE)) were probed with chitosan modified AFM tips and the corresponding desorption energies, calculated from plateau-like features, were attributed to the desorption of individual polymer strands. Desorption energies of 2.0±0.3×10(-20)J, 1.8±0.3×10(-20)J and 3.5±0.3×10(-20)J were obtained for glass, SAM of mercaptoundecanoic/dodecanethiol and PTFE, respectively. These single molecule level results can be used as a basis for investigating chitosan and chitosan-based materials for biomaterial applications. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

    Neuman, Keir C.; Nagy, Attila

    2012-01-01

    Single-molecule force spectroscopy has emerged as a powerful tool to investigate the forces and motions associated with biological molecules and enzymatic activity. The most common force spectroscopy techniques are optical tweezers, magnetic tweezers and atomic force microscopy. These techniques are described and illustrated with examples highlighting current capabilities and limitations. PMID:18511917

  15. Statistical filtering in fluorescence microscopy and fluorescence correlation spectroscopy

    Macháň, Radek; Kapusta, Peter; Hof, Martin

    Roč. 406 , č. 20 (2014), s. 4797-4813 ISSN 1618-2642 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : Filtered fluorescence correlation spectroscopy * Fluorescence lifetime correlation spectroscopy * Fluorescence spectral correlation spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.436, year: 2014

  16. Obesity-related differences in neural correlates of force control.

    Mehta, Ranjana K; Shortz, Ashley E

    2014-01-01

    Greater body segment mass due to obesity has shown to impair gross and fine motor functions and reduce balance control. While recent studies suggest that obesity may be linked with altered brain functions involved in fine motor tasks, this association is not well investigated. The purpose of this study was to examine the neural correlates of motor performance in non-obese and obese adults during force control of two upper extremity muscles. Nine non-obese and eight obese young adults performed intermittent handgrip and elbow flexion exertions at 30% of their respective muscle strengths for 4 min. Functional near infrared spectroscopy was employed to measure neural activity in the prefrontal cortex bilaterally, joint steadiness was computed using force fluctuations, and ratings of perceived exertions (RPEs) were obtained to assess perceived effort. Obesity was associated with higher force fluctuations and lower prefrontal cortex activation during handgrip exertions, while RPE scores remained similar across both groups. No obesity-related differences in neural activity, force fluctuation, or RPE scores were observed during elbow flexion exertions. The study is one of the first to examine obesity-related differences on prefrontal cortex activation during force control of the upper extremity musculature. The study findings indicate that the neural correlates of motor activity in the obese may be muscle-specific. Future work is warranted to extend the investigation to monitoring multiple motor-function related cortical regions and examining obesity differences with different task parameters (e.g., longer duration, increased precision demands, larger muscles, etc.).

  17. Quantifying Young's moduli of protein fibrils and particles with bimodal force spectroscopy.

    Gilbert, Jay; Charnley, Mirren; Cheng, Christopher; Reynolds, Nicholas P; Jones, Owen G

    2017-10-19

    Force spectroscopy is a means of obtaining mechanical information of individual nanometer-scale structures in composite materials, such as protein assemblies for use in consumer films or gels. As a recently developed force spectroscopy technique, bimodal force spectroscopy relates frequency shifts in cantilevers simultaneously excited at multiple frequencies to the elastic properties of the contacted material, yet its utility for quantitative characterization of biopolymer assemblies has been limited. In this study, a linear correlation between experimental frequency shift and Young's modulus of polymer films was used to calibrate bimodal force spectroscopy and quantify Young's modulus of two protein nanostructures: β-lactoglobulin fibrils and zein nanoparticles. Cross-sectional Young's modulus of protein fibrils was determined to be 1.6 GPa while the modulus of zein nanoparticles was determined as 854 MPa. Parallel measurement of β-lactoglobulin fibril by a competing pulsed-force technique found a higher cross-sectional Young's modulus, highlighting the importance of comparative calibration against known standards in both pulsed and bimodal force spectroscopies. These findings demonstrate a successful procedure for measuring mechanical properties of individual protein assemblies with potential use in biological or packaging applications using bimodal force spectroscopy.

  18. Force-detected nanoscale absorption spectroscopy in water at room temperature using an optical trap

    Parobek, Alexander; Black, Jacob W.; Kamenetska, Maria; Ganim, Ziad

    2018-04-01

    Measuring absorption spectra of single molecules presents a fundamental challenge for standard transmission-based instruments because of the inherently low signal relative to the large background of the excitation source. Here we demonstrate a new approach for performing absorption spectroscopy in solution using a force measurement to read out optical excitation at the nanoscale. The photoinduced force between model chromophores and an optically trapped gold nanoshell has been measured in water at room temperature. This photoinduced force is characterized as a function of wavelength to yield the force spectrum, which is shown to be correlated to the absorption spectrum for four model systems. The instrument constructed for these measurements combines an optical tweezer with frequency domain absorption spectroscopy over the 400-800 nm range. These measurements provide proof-of-principle experiments for force-detected nanoscale spectroscopies that operate under ambient chemical conditions.

  19. Analysis of DNA interactions using single-molecule force spectroscopy.

    Ritzefeld, Markus; Walhorn, Volker; Anselmetti, Dario; Sewald, Norbert

    2013-06-01

    Protein-DNA interactions are involved in many biochemical pathways and determine the fate of the corresponding cell. Qualitative and quantitative investigations on these recognition and binding processes are of key importance for an improved understanding of biochemical processes and also for systems biology. This review article focusses on atomic force microscopy (AFM)-based single-molecule force spectroscopy and its application to the quantification of forces and binding mechanisms that lead to the formation of protein-DNA complexes. AFM and dynamic force spectroscopy are exciting tools that allow for quantitative analysis of biomolecular interactions. Besides an overview on the method and the most important immobilization approaches, the physical basics of the data evaluation is described. Recent applications of AFM-based force spectroscopy to investigate DNA intercalation, complexes involving DNA aptamers and peptide- and protein-DNA interactions are given.

  20. Charge-state dynamics in electrostatic force spectroscopy

    Ondráček, Martin; Hapala, Prokop; Jelínek, Pavel

    2016-01-01

    Roč. 27, č. 27 (2016), 1-13, č. článku 274005. ISSN 0957-4484 R&D Projects: GA ČR(CZ) GA14-02079S Institutional support: RVO:68378271 Keywords : atomic force microscopy * electron tunneling * redox nanoswitches * electrostatic force spectroscopy Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 3.440, year: 2016

  1. Spectroscopy and atomic force microscopy of biomass.

    Tetard, L; Passian, A; Farahi, R H; Kalluri, U C; Davison, B H; Thundat, T

    2010-05-01

    Scanning probe microscopy has emerged as a powerful approach to a broader understanding of the molecular architecture of cell walls, which may shed light on the challenge of efficient cellulosic ethanol production. We have obtained preliminary images of both Populus and switchgrass samples using atomic force microscopy (AFM). The results show distinctive features that are shared by switchgrass and Populus. These features may be attributable to the lignocellulosic cell wall composition, as the collected images exhibit the characteristic macromolecular globule structures attributable to the lignocellulosic systems. Using both AFM and a single case of mode synthesizing atomic force microscopy (MSAFM) to characterize Populus, we obtained images that clearly show the cell wall structure. The results are of importance in providing a better understanding of the characteristic features of both mature cells as well as developing plant cells. In addition, we present spectroscopic investigation of the same samples.

  2. Noise Spectroscopy in Strongly Correlated Oxides

    Alsaqqa, Ali M.

    Strongly correlated materials are an interesting class of materials, thanks to the novel electronic and magnetic phenomena they exhibit as a result of the interplay of various degrees of freedom. This gives rise to an array of potential applications, from Mott-FET to magnetic storage. Many experimental probes have been used to study phase transitions in strongly correlated oxides. Among these, resistance noise spectroscopy, together with conventional transport measurements, provides a unique viewpoint to understand the microscopic dynamics near the phase transitions in these oxides. In this thesis, utilizing noise spectroscopy and transport measurements, four different strongly correlated materials were studied: (1) neodymium nickel oxide (NdNiO 3) ultrathin films, (2) vanadium dioxide (VO2) microribbons, (3) copper vanadium bronze (CuxV2O 5) microribbons and (4) niobium triselenide (NbSe3) microribbons. Ultra thin films of rare-earth nickelates exhibit several temperature-driven phase transitions. In this thesis, we studied the metal-insulator and Neel transitions in a series of NdNiO3 films with different lattice mismatches. Upon colling down, the metal-insulator phase transition is accompanied by a structural (orthorohombic to monoclinic) and magnetic (paramagnetic to antiferromagnetic) transitions as well, making the problem more interesting and complex at the same time. The noise is of the 1/f type and is Gaussian in the high temperature phase, however deviations are seen in the low temperature phases. Below the metal-insulator transition, noise magnitude increases by orders of magnitude: a sign of inhomogeneous electrical conduction as result of phase separation. This is further assured by the non-Gaussian noise signature. At very low temperatures (T switches between Gaussian and non-Gaussian over several hours, possibly arising from dynamically competing ground states. VO2 is one of the most widely studied strongly correlated oxides and is important from the

  3. Compressive Force Spectroscopy: From Living Cells to Single Proteins.

    Wang, Jiabin; Liu, Meijun; Shen, Yi; Sun, Jielin; Shao, Zhifeng; Czajkowsky, Daniel Mark

    2018-03-23

    One of the most successful applications of atomic force microscopy (AFM) in biology involves monitoring the effect of force on single biological molecules, often referred to as force spectroscopy. Such studies generally entail the application of pulling forces of different magnitudes and velocities upon individual molecules to resolve individualistic unfolding/separation pathways and the quantification of the force-dependent rate constants. However, a less recognized variation of this method, the application of compressive force, actually pre-dates many of these "tensile" force spectroscopic studies. Further, beyond being limited to the study of single molecules, these compressive force spectroscopic investigations have spanned samples as large as living cells to smaller, multi-molecular complexes such as viruses down to single protein molecules. Correspondingly, these studies have enabled the detailed characterization of individual cell states, subtle differences between seemingly identical viral structures, as well as the quantification of rate constants of functionally important, structural transitions in single proteins. Here, we briefly review some of the recent achievements that have been obtained with compressive force spectroscopy using AFM and highlight exciting areas of its future development.

  4. Optical Spectroscopy and Imaging of Correlated Spin Orbit Phases

    2016-06-14

    Unlimited UU UU UU UU 14-06-2016 15-Mar-2013 14-Mar-2016 Final Report: Optical Spectroscopy and Imaging of Correlated Spin-Orbit Phases The views...Box 12211 Research Triangle Park, NC 27709-2211 Ultrafast optical spectroscopy , nonlinear optical spectroscopy , iridates, cuprates REPORT...California Blvd. Pasadena, CA 91125 -0001 ABSTRACT Number of Papers published in peer-reviewed journals: Final Report: Optical Spectroscopy and

  5. Two-focus fluorescence correlation spectroscopy

    Dertinger, T.

    2007-05-01

    Fluorescence Correlation Spectroscopy (FCS) has been invented more than 30 years ago and experienced a renaissance after stable and affordable laser sources and low-noise single-photon detectors have become available. Its ability to measure diffusion coefficients at nanomolar concentrations of analyte made it a widely used tool in biophysics. However, in recent years it has been shown by many authors that aberrational (e.g. astigmatism) and photophysical effects (e.g. optical saturation) may influence the result of an FCS experiment dramatically, so that a precise and reliable estimation of the diffusion coefficient is no longer possible. In this thesis, we report on the development, implementation, and application of a new and robust modification of FCS that we termed two-focus FCS (2fFCS) and which fulfils two requirements: (i) It introduces an external ruler into the measurement by generating two overlapping laser foci of precisely known and fixed distance. (ii) These two foci and corresponding detection regions are generated in such a way that the corresponding molecule detection functions (MDFs) are sufficiently well described by a simple two-parameter model yielding accurate diffusion coefficients when applied to 2fFCS data analysis. Both these properties enable us to measure absolute values of the diffusion coefficient with an accuracy of a few percent. Moreover, it will turn out that the new technique is robust against refractive index mismatch, coverslide thickness deviations, and optical saturation effects, which so often trouble conventional FCS measurements. This thesis deals mainly with the introduction of the new measurement scheme, 2fFCS, but also presents several applications with far-reaching importance. (orig.)

  6. Two-focus fluorescence correlation spectroscopy

    Dertinger, T.

    2007-05-15

    Fluorescence Correlation Spectroscopy (FCS) has been invented more than 30 years ago and experienced a renaissance after stable and affordable laser sources and low-noise single-photon detectors have become available. Its ability to measure diffusion coefficients at nanomolar concentrations of analyte made it a widely used tool in biophysics. However, in recent years it has been shown by many authors that aberrational (e.g. astigmatism) and photophysical effects (e.g. optical saturation) may influence the result of an FCS experiment dramatically, so that a precise and reliable estimation of the diffusion coefficient is no longer possible. In this thesis, we report on the development, implementation, and application of a new and robust modification of FCS that we termed two-focus FCS (2fFCS) and which fulfils two requirements: (i) It introduces an external ruler into the measurement by generating two overlapping laser foci of precisely known and fixed distance. (ii) These two foci and corresponding detection regions are generated in such a way that the corresponding molecule detection functions (MDFs) are sufficiently well described by a simple two-parameter model yielding accurate diffusion coefficients when applied to 2fFCS data analysis. Both these properties enable us to measure absolute values of the diffusion coefficient with an accuracy of a few percent. Moreover, it will turn out that the new technique is robust against refractive index mismatch, coverslide thickness deviations, and optical saturation effects, which so often trouble conventional FCS measurements. This thesis deals mainly with the introduction of the new measurement scheme, 2fFCS, but also presents several applications with far-reaching importance. (orig.)

  7. Charge-Transfer Complexes Studied by Dynamic Force Spectroscopy

    Jurriaan Huskens

    2013-03-01

    Full Text Available In this paper, the strength and kinetics of two charge-transfer complexes, naphthol-methylviologen and pyrene-methylviologen, are studied using dynamic force spectroscopy. The dissociation rates indicate an enhanced stability of the pyrene-methylviologen complex, which agrees with its higher thermodynamic stability compared to naphthol-methylviologen complex.

  8. Investigation of specific interactions between T7 promoter and T7 RNA polymerase by force spectroscopy using atomic force microscope.

    Zhang, Xiaojuan; Yao, Zhixuan; Duan, Yanting; Zhang, Xiaomei; Shi, Jinsong; Xu, Zhenghong

    2018-01-11

    The specific recognition and binding of promoter and RNA polymerase is the first step of transcription initiation in bacteria and largely determines transcription activity. Therefore, direct analysis of the interaction between promoter and RNA polymerase in vitro may be a new strategy for promoter characterization, to avoid interference due to the cell's biophysical condition and other regulatory elements. In the present study, the specific interaction between T7 promoter and T7 RNA polymerase was studied as a model system using force spectroscopy based on atomic force microscope (AFM). The specific interaction between T7 promoter and T7 RNA polymerase was verified by control experiments, and the rupture force in this system was measured as 307.2 ± 6.7 pN. The binding between T7 promoter mutants with various promoter activities and T7 RNA polymerase was analyzed. Interaction information including rupture force, rupture distance and binding percentage were obtained in vitro , and reporter gene expression regulated by these promoters was also measured according to a traditional promoter activity characterization method in vivo Using correlation analysis, it was found that the promoter strength characterized by reporter gene expression was closely correlated with rupture force and the binding percentage by force spectroscopy. These results indicated that the analysis of the interaction between promoter and RNA polymerase using AFM-based force spectroscopy was an effective and valid approach for the quantitative characterization of promoters. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  9. Fluorescence Lifetime Correlation Spectroscopy (FLCS): Concepts, Applications and Outlook

    Kapusta, Peter; Macháň, Radek; Benda, A.; Hof, Martin

    2012-01-01

    Roč. 13, č. 10 (2012), s. 12890-12910 E-ISSN 1422-0067 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : fluorescence correlation spectroscopy (FCS) * time correlated single photon counting (TCSPC) * fluorescence cross-correlation spectroscopy (FCCS) Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.464, year: 2012

  10. Athermalization in atomic force microscope based force spectroscopy using matched microstructure coupling.

    Torun, H; Finkler, O; Degertekin, F L

    2009-07-01

    The authors describe a method for athermalization in atomic force microscope (AFM) based force spectroscopy applications using microstructures that thermomechanically match the AFM probes. The method uses a setup where the AFM probe is coupled with the matched structure and the displacements of both structures are read out simultaneously. The matched structure displaces with the AFM probe as temperature changes, thus the force applied to the sample can be kept constant without the need for a separate feedback loop for thermal drift compensation, and the differential signal can be used to cancel the shift in zero-force level of the AFM.

  11. Minimizing pulling geometry errors in atomic force microscope single molecule force spectroscopy.

    Rivera, Monica; Lee, Whasil; Ke, Changhong; Marszalek, Piotr E; Cole, Daniel G; Clark, Robert L

    2008-10-01

    In atomic force microscopy-based single molecule force spectroscopy (AFM-SMFS), it is assumed that the pulling angle is negligible and that the force applied to the molecule is equivalent to the force measured by the instrument. Recent studies, however, have indicated that the pulling geometry errors can drastically alter the measured force-extension relationship of molecules. Here we describe a software-based alignment method that repositions the cantilever such that it is located directly above the molecule's substrate attachment site. By aligning the applied force with the measurement axis, the molecule is no longer undergoing combined loading, and the full force can be measured by the cantilever. Simulations and experimental results verify the ability of the alignment program to minimize pulling geometry errors in AFM-SMFS studies.

  12. Single molecule force spectroscopy: methods and applications in biology

    Shen Yi; Hu Jun

    2012-01-01

    Single molecule measurements have transformed our view of biomolecules. Owing to the ability of monitoring the activity of individual molecules, we now see them as uniquely structured, fluctuating molecules that stochastically transition between frequently many substrates, as two molecules do not follow precisely the same trajectory. Indeed, it is this discovery of critical yet short-lived substrates that were often missed in ensemble measurements that has perhaps contributed most to the better understanding of biomolecular functioning resulting from single molecule experiments. In this paper, we give a review on the three major techniques of single molecule force spectroscopy, and their applications especially in biology. The single molecular study of biotin-streptavidin interactions is introduced as a successful example. The problems and prospects of the single molecule force spectroscopy are discussed, too. (authors)

  13. Single-cell force spectroscopy of pili-mediated adhesion

    Sullan, Ruby May A.; Beaussart, Audrey; Tripathi, Prachi; Derclaye, Sylvie; El-Kirat-Chatel, Sofiane; Li, James K.; Schneider, Yves-Jacques; Vanderleyden, Jos; Lebeer, Sarah; Dufrêne, Yves F.

    2013-12-01

    Although bacterial pili are known to mediate cell adhesion to a variety of substrates, the molecular interactions behind this process are poorly understood. We report the direct measurement of the forces guiding pili-mediated adhesion, focusing on the medically important probiotic bacterium Lactobacillus rhamnosus GG (LGG). Using non-invasive single-cell force spectroscopy (SCFS), we quantify the adhesion forces between individual bacteria and biotic (mucin, intestinal cells) or abiotic (hydrophobic monolayers) surfaces. On hydrophobic surfaces, bacterial pili strengthen adhesion through remarkable nanospring properties, which - presumably - enable the bacteria to resist high shear forces under physiological conditions. On mucin, nanosprings are more frequent and adhesion forces larger, reflecting the influence of specific pili-mucin bonds. Interestingly, these mechanical responses are no longer observed on human intestinal Caco-2 cells. Rather, force curves exhibit constant force plateaus with extended ruptures reflecting the extraction of membrane nanotethers. These single-cell analyses provide novel insights into the molecular mechanisms by which piliated bacteria colonize surfaces (nanosprings, nanotethers), and offer exciting avenues in nanomedicine for understanding and controlling the adhesion of microbial cells (probiotics, pathogens).

  14. Atomic force microscopy and force spectroscopy on the assessment of protein folding and functionality.

    Carvalho, Filomena A; Martins, Ivo C; Santos, Nuno C

    2013-03-01

    Atomic force microscopy (AFM) applied to biological systems can, besides generating high-quality and well-resolved images, be employed to study protein folding via AFM-based force spectroscopy. This approach allowed remarkable advances in the measurement of inter- and intramolecular interaction forces with piconewton resolution. The detection of specific interaction forces between molecules based on the AFM sensitivity and the manipulation of individual molecules greatly advanced the understanding of intra-protein and protein-ligand interactions. Apart from the academic interest in the resolution of basic scientific questions, this technique has also key importance on the clarification of several biological questions of immediate biomedical relevance. Force spectroscopy is an especially appropriate technique for "mechanical proteins" that can provide crucial information on single protein molecules and/or domains. Importantly, it also has the potential of combining in a single experiment spatial and kinetic measurements. Here, the main principles of this methodology are described, after which the ability to measure interactions at the single-molecule level is discussed, in the context of relevant protein-folding examples. We intend to demonstrate the potential of AFM-based force spectroscopy in the study of protein folding, especially since this technique is able to circumvent some of the difficulties typically encountered in classical thermal/chemical denaturation studies. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. 25 ns software correlator for photon and fluorescence correlation spectroscopy

    Magatti, Davide; Ferri, Fabio

    2003-02-01

    A 25 ns time resolution, multi-tau software correlator developed in LABVIEW based on the use of a standard photon counting unit, a fast timer/counter board (6602-PCI National Instrument) and a personal computer (PC) (1.5 GHz Pentium 4) is presented and quantitatively discussed. The correlator works by processing the stream of incoming data in parallel according to two different algorithms: For large lag times (τ⩾100 μs), a classical time-mode (TM) scheme, based on the measure of the number of pulses per time interval, is used; differently, for τ⩽100 μs a photon-mode (PM) scheme is adopted and the time sequence of the arrival times of the photon pulses is measured. By combining the two methods, we developed a system capable of working out correlation functions on line, in full real time for the TM correlator and partially in batch processing for the PM correlator. For the latter one, the duty cycle depends on the count rate of the incoming pulses, being ˜100% for count rates ⩽3×104 Hz, ˜15% at 105 Hz, and ˜1% at 106 Hz. For limitations imposed by the fairly small first-in, first-out (FIFO) buffer available on the counter board, the maximum count rate permissible for a proper functioning of the PM correlator is limited to ˜105 Hz. However, this limit can be removed by using a board with a deeper FIFO. Similarly, the 25 ns time resolution is only limited by maximum clock frequency available on the 6602-PCI and can be easily improved by using a faster clock. When tested on dilute solutions of calibrated latex spheres, the overall performances of the correlator appear to be comparable with those of commercial hardware correlators, but with several nontrivial advantages related to its flexibility, low cost, and easy adaptability to future developments of PC and data acquisition technology.

  16. Functionalization of gold and nanocrystalline diamond atomic force microscope tips for single molecule force spectroscopy

    Drew, Michael E.

    The atomic force microscope (AFM) has fueled interest in nanotechnology because of its ability to image surfaces at the nanometer level and act as a molecular force sensor. Functionalization of the surface of an AFM tip surface in a stable, controlled manner expands the capabilities of the AFM and enables additional applications in the fields of single molecule force spectroscopy and nanolithography. Two AFM tip functionalizations are described: the assembly of tripodal molecular tips onto gold AFM tips and the photochemical attachment of terminal alkenes to nanocrystalline diamond (NCD) AFM tips. Two separate tripodal molecules with different linker lengths and a monopodal molecule terminated with biotin were synthesized to attach to a gold AFM tip for single molecule force spectroscopy. The immobilization of these molecules was examined by contact angle measurements, spectroscopic ellipsometry, infrared, and near edge x-ray absorption fine structure (NEXAFS) spectroscopy. All three molecules displayed rupture forces that agreed with previously reported values for the biotin--avidin rupture. The tripodal molecular tip displayed narrower distribution in their force histograms than the monopodal molecular tip. The performance of the tripodal molecular tip was compared to the monopodal molecular tip in single molecule force spectroscopy studies. Over repeated measurements, the distribution of forces for the monopodal molecular tip shifted to lower forces, whereas the distribution for the tripodal molecular tip remained constant throughout. Loading rate dependence and control experiments further indicated that the rupture forces of the tripod molecular tips were specific to the biotin--NeutrAvidin interaction. The second functionalization method used the photochemical attachment of undecylenic acid to NCD AFM tips. The photochemical attachment of undecylenic acid to hydrogen-terminated NCD wafer surfaces was investigated by contact angle measurements, x

  17. Disability correlates in Canadian Armed Forces Regular Force Veterans.

    Thompson, James M; Pranger, Tina; Sweet, Jill; VanTil, Linda; McColl, Mary Ann; Besemann, Markus; Shubaly, Colleen; Pedlar, David

    2015-01-01

    This study was undertaken to inform disability mitigation for military veterans by identifying personal, environmental, and health factors associated with activity limitations. A sample of 3154 Canadian Armed Forces Regular Force Veterans who were released during 1998-2007 participated in the 2010 Survey on Transition to Civilian Life. Associations between personal and environmental factors, health conditions and activity limitations were explored using ordinal logistic regression. The prevalence of activity reduction in life domains was higher than the Canadian general population (49% versus 21%), as was needing assistance with at least one activity of daily living (17% versus 5%). Prior to adjusting for health conditions, disability odds were elevated for increased age, females, non-degree post-secondary graduation, low income, junior non-commissioned members, deployment, low social support, low mastery, high life stress, and weak sense of community belonging. Reduced odds were found for private/recruit ranks. Disability odds were highest for chronic pain (10.9), any mental health condition (2.7), and musculoskeletal conditions (2.6), and there was a synergistic additive effect of physical and mental health co-occurrence. Disability, measured as activity limitation, was associated with a range of personal and environmental factors and health conditions, indicating multifactorial and multidisciplinary approaches to disability mitigation.

  18. Probing living bacterial adhesion by single cell force spectroscopy using atomic force microscopy

    Zeng, Guanghong; Ogaki, Ryosuke; Regina, Viduthalai R.

    be considered. We have therefore developed a simple and versatile method to make single-cell bacterial probes for measuring single cell adhesion with atomic force microscopy (AFM).[1] A single-cell probe was readily made by picking up a bacterial cell from a glass surface using a tipless AFM cantilever coated...... random immobilization is obtained by submerging the cantilever in a bacterial suspension. The reported method provides a general platform for investigating single cell interactions of bacteria with different surfaces and other cells by AFM force spectroscopy, thus improving our understanding....... The strain-dependent susceptibility to bacterial colonization on conventional PLL-g-PEG illustrates how bacterial diversity challenges development of “universal” antifouling coatings, and AFM single-cell force spectroscopy was proven to be a powerful tool to provide insights into the molecular mechanisms...

  19. Hooke: an open software platform for force spectroscopy.

    Sandal, Massimo; Benedetti, Fabrizio; Brucale, Marco; Gomez-Casado, Alberto; Samorì, Bruno

    2009-06-01

    Hooke is an open source, extensible software intended for analysis of atomic force microscope (AFM)-based single molecule force spectroscopy (SMFS) data. We propose it as a platform on which published and new algorithms for SMFS analysis can be integrated in a standard, open fashion, as a general solution to the current lack of a standard software for SMFS data analysis. Specific features and support for file formats are coded as independent plugins. Any user can code new plugins, extending the software capabilities. Basic automated dataset filtering and semi-automatic analysis facilities are included. Software and documentation are available at (http://code.google.com/p/hooke). Hooke is a free software under the GNU Lesser General Public License.

  20. Medical applications of atomic force microscopy and Raman spectroscopy.

    Choi, Samjin; Jung, Gyeong Bok; Kim, Kyung Sook; Lee, Gi-Ja; Park, Hun-Kuk

    2014-01-01

    This paper reviews the recent research and application of atomic force microscopy (AFM) and Raman spectroscopy techniques, which are considered the multi-functional and powerful toolkits for probing the nanostructural, biomechanical and physicochemical properties of biomedical samples in medical science. We introduce briefly the basic principles of AFM and Raman spectroscopy, followed by diagnostic assessments of some selected diseases in biomedical applications using them, including mitochondria isolated from normal and ischemic hearts, hair fibers, individual cells, and human cortical bone. Finally, AFM and Raman spectroscopy applications to investigate the effects of pharmacotherapy, surgery, and medical device therapy in various medicines from cells to soft and hard tissues are discussed, including pharmacotherapy--paclitaxel on Ishikawa and HeLa cells, telmisartan on angiotensin II, mitomycin C on strabismus surgery and eye whitening surgery, and fluoride on primary teeth--and medical device therapy--collagen cross-linking treatment for the management of progressive keratoconus, radiofrequency treatment for skin rejuvenation, physical extracorporeal shockwave therapy for healing of Achilles tendinitis, orthodontic treatment, and toothbrushing time to minimize the loss of teeth after exposure to acidic drinks.

  1. Image correlation spectroscopy: mapping correlations in space, time, and reciprocal space.

    Wiseman, Paul W

    2013-01-01

    This chapter presents an overview of two recent implementations of image correlation spectroscopy (ICS). The background theory is presented for spatiotemporal image correlation spectroscopy and image cross-correlation spectroscopy (STICS and STICCS, respectively) as well as k-(reciprocal) space image correlation spectroscopy (kICS). An introduction to the background theory is followed by sections outlining procedural aspects for properly implementing STICS, STICCS, and kICS. These include microscopy image collection, sampling in space and time, sample and fluorescent probe requirements, signal to noise, and background considerations that are all required to properly implement the ICS methods. Finally, procedural steps for immobile population removal and actual implementation of the ICS analysis programs to fluorescence microscopy image time stacks are described. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Probing of miniPEGγ-PNA-DNA Hybrid Duplex Stability with AFM Force Spectroscopy.

    Dutta, Samrat; Armitage, Bruce A; Lyubchenko, Yuri L

    2016-03-15

    Peptide nucleic acids (PNA) are synthetic polymers, the neutral peptide backbone of which provides elevated stability to PNA-PNA and PNA-DNA hybrid duplexes. It was demonstrated that incorporation of diethylene glycol (miniPEG) at the γ position of the peptide backbone increased the thermal stability of the hybrid duplexes (Sahu, B. et al. J. Org. Chem. 2011, 76, 5614-5627). Here, we applied atomic force microscopy (AFM) based single molecule force spectroscopy and dynamic force spectroscopy (DFS) to test the strength and stability of the hybrid 10 bp duplex. This hybrid duplex consisted of miniPEGγ-PNA and DNA of the same length (γ(MP)PNA-DNA), which we compared to a DNA duplex with a homologous sequence. AFM force spectroscopy data obtained at the same conditions showed that the γ(MP)PNA-DNA hybrid is more stable than the DNA counterpart, 65 ± 15 pN vs 47 ± 15 pN, respectively. The DFS measurements performed in a range of pulling speeds analyzed in the framework of the Bell-Evans approach yielded a dissociation constant, koff ≈ 0.030 ± 0.01 s⁻¹ for γ(MP)PNA-DNA hybrid duplex vs 0.375 ± 0.18 s⁻¹ for the DNA-DNA duplex suggesting that the hybrid duplex is much more stable. Correlating the high affinity of γ(MP)PNA-DNA to slow dissociation kinetics is consistent with prior bulk characterization by surface plasmon resonance. Given the growing interest in γ(MP)PNA as well as other synthetic DNA analogues, the use of single molecule experiments along with computational analysis of force spectroscopy data will provide direct characterization of various modifications as well as higher order structures such as triplexes and quadruplexes.

  3. Effects of solute--solvent attractive forces on hydrophobic correlations

    Pratt, L.R.; Chandler, D.

    1980-01-01

    A theory is presented for the effect of slowly varying attractive forces on correlations between nonpolar solutes in dilute aqueous solution. We find that hydrophobic correlations are sensitive to relatively long range slowly varying interactions. Thus, it is possible to make qualitative changes in these correlations by introducing small changes in the attractive forces. Several model calculations are presented to illustrate these facts. The contributions of the Lennard-Jones attractive forces to the computer simulation results of Pangali, Rao, and Berne are calculated. For this case it is found that the potential of mean force between spherical nonpolar solutes is hardly affected by inclusion of attractive forces. However, the osmotic second virial coefficient is dominated by the contributions of the attractive forces. For spherical solutes which provide a reasonable model for the methane molecule, inclusion of attractive forces produces a qualitative change in the methane--methane potential of mean force. The connection between these effects of slowly varying attractive forces and the enthalpic part of Ben-Naim's deltaA/sup H/I is discussed

  4. Taking nanomedicine teaching into practice with atomic force microscopy and force spectroscopy.

    Carvalho, Filomena A; Freitas, Teresa; Santos, Nuno C

    2015-12-01

    Atomic force microscopy (AFM) is a useful and powerful tool to study molecular interactions applied to nanomedicine. The aim of the present study was to implement a hands-on atomic AFM course for graduated biosciences and medical students. The course comprises two distinct practical sessions, where students get in touch with the use of an atomic force microscope by performing AFM scanning images of human blood cells and force spectroscopy measurements of the fibrinogen-platelet interaction. Since the beginning of this course, in 2008, the overall rating by the students was 4.7 (out of 5), meaning a good to excellent evaluation. Students were very enthusiastic and produced high-quality AFM images and force spectroscopy data. The implementation of the hands-on AFM course was a success, giving to the students the opportunity of contact with a technique that has a wide variety of applications on the nanomedicine field. In the near future, nanomedicine will have remarkable implications in medicine regarding the definition, diagnosis, and treatment of different diseases. AFM enables students to observe single molecule interactions, enabling the understanding of molecular mechanisms of different physiological and pathological processes at the nanoscale level. Therefore, the introduction of nanomedicine courses in bioscience and medical school curricula is essential. Copyright © 2015 The American Physiological Society.

  5. Distribution of diffusion times determined by fluorescence (lifetime) correlation spectroscopy

    Pánek, Jiří; Loukotová, Lenka; Hrubý, Martin; Štěpánek, Petr

    2018-01-01

    Roč. 51, č. 8 (2018), s. 2796-2804 ISSN 0024-9297 R&D Projects: GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : polymer solution * fluorescence correlation spectroscopy * diffusion time distribution Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 5.835, year: 2016

  6. Investigating single molecule adhesion by atomic force spectroscopy.

    Stetter, Frank W S; Kienle, Sandra; Krysiak, Stefanie; Hugel, Thorsten

    2015-02-27

    Atomic force spectroscopy is an ideal tool to study molecules at surfaces and interfaces. An experimental protocol to couple a large variety of single molecules covalently onto an AFM tip is presented. At the same time the AFM tip is passivated to prevent unspecific interactions between the tip and the substrate, which is a prerequisite to study single molecules attached to the AFM tip. Analyses to determine the adhesion force, the adhesion length, and the free energy of these molecules on solid surfaces and bio-interfaces are shortly presented and external references for further reading are provided. Example molecules are the poly(amino acid) polytyrosine, the graft polymer PI-g-PS and the phospholipid POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine). These molecules are desorbed from different surfaces like CH3-SAMs, hydrogen terminated diamond and supported lipid bilayers under various solvent conditions. Finally, the advantages of force spectroscopic single molecule experiments are discussed including means to decide if truly a single molecule has been studied in the experiment.

  7. Orthodontic Force Application in Correlation with Salivary Lactate Dehydrogenase Activity

    Erik Husin

    2013-07-01

    Full Text Available Orthodontic tooth movement generate mechanical forces to periodontal ligament and alveolar bone. The forces correlate with initial responses of periodontal tissues and involving many metabolic changes. One of the metabolic changes detected in saliva is lactate dehydrogenase (LDH activity. Objectives: To evaluate the correlation between orthodontic interrupted force application, lactate dehydrogenase activity and the distance of tooth movement. Methods: upper premolar, pre-retraction of upper canine and 1, 7, 14, 21 and 28 days post-retraction of upper canine with 100g interrupted orthodontic force. Results: duration of force (F=11.926 p 14 and 28 days post-retraction of canine. The region of retraction correlated with the distance of tooth movement (F=7.377 p=0.007. The duration of force correlated with the distance of tooth movement (F=66.554 p=0.000. retraction of canine. Conclusion: This study concluded that orthodontic interrupted force application on canine could increase the distance of tooth movement and LDH activity in saliva.

  8. Fluorescence Correlation Spectroscopy Using Octadecylrhodamine B as a Specific Micelle-Binding Fluorescent Tag, Light Scattering and Tapping Mode Atomic Force Microscopy Studies of Amphiphilic Water-Soluble Block Copolymer Micelles

    Humpolíčková, J.; Procházka, K.; Hof, Martin; Tuzar, Zdeněk; Špírková, Milena

    2003-01-01

    Roč. 19, - (2003), s. 4111-4119 ISSN 0743-7463 R&D Projects: GA MŠk LN00A032; GA ČR GA203/01/0536; GA ČR GA203/01/0735 Institutional research plan: CEZ:AV0Z4050913; CEZ:AV0Z4040901 Keywords : fluorescence * spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.098, year: 2003

  9. Advancements of two dimensional correlation spectroscopy in protein researches

    Tao, Yanchun; Wu, Yuqing; Zhang, Liping

    2018-05-01

    The developments of two-dimensional correlation spectroscopy (2DCOS) applications in protein studies are discussed, especially for the past two decades. The powerful utilities of 2DCOS combined with various analytical techniques in protein studies are summarized. The emphasis is on the vibration spectroscopic techniques including IR, NIR, Raman and optical activity (ROA), as well as vibration circular dichroism (VCD) and fluorescence spectroscopy. In addition, some new developments, such as hetero-spectral 2DCOS, moving-window correlation, and model based correlation, are also reviewed for their utility in the investigation of the secondary structure, denaturation, folding and unfolding changes of protein. Finally, the new possibility and challenges of 2DCOS in protein research are highlighted as well.

  10. Probing new light force-mediators by isotope shift spectroscopy

    Berengut, Julian C.; Budker, Dmitry; California Univ., Berkeley, CA; Lawrence Berkeley National Laboratory, Berkeley, CA; Delaunay, Cedric

    2017-04-01

    In this Letter we explore the potential of probing new light force-carriers, with spin-independent couplings to the electron and the neutron, using precision isotope shift spectroscopy. We develop a formalism to interpret linear King plots as bounds on new physics with minimal theory inputs. We focus only on bounding the new physics contributions that can be calculated independently of the Standard Model nuclear effects. We apply our method to existing Ca"+ data and project its sensitivity to possibly existing new bosons using narrow transitions in other atoms and ions (specifically, Sr and Yb). Future measurements are expected to improve the relative precision by five orders of magnitude, and can potentially lead to an unprecedented sensitivity for bosons within the 10 keV to 10 MeV mass range.

  11. Probing new light force-mediators by isotope shift spectroscopy

    Berengut, Julian C. [New South Wales Univ., Sydney, NSW (Australia). School of Physics; Budker, Dmitry [Mainz Univ. (Germany). Helmholtz-Inst. Mainz; California Univ., Berkeley, CA (United States). Physics Dept.; Lawrence Berkeley National Laboratory, Berkeley, CA (United States). Nuclear Science Div.; Delaunay, Cedric [Savoie Mont Blanc Univ., Annecy-le-Vieux (France). Laboratoire d' Annecy-le-Vieux de Physique Theorique LAPTh; and others

    2017-04-15

    In this Letter we explore the potential of probing new light force-carriers, with spin-independent couplings to the electron and the neutron, using precision isotope shift spectroscopy. We develop a formalism to interpret linear King plots as bounds on new physics with minimal theory inputs. We focus only on bounding the new physics contributions that can be calculated independently of the Standard Model nuclear effects. We apply our method to existing Ca{sup +} data and project its sensitivity to possibly existing new bosons using narrow transitions in other atoms and ions (specifically, Sr and Yb). Future measurements are expected to improve the relative precision by five orders of magnitude, and can potentially lead to an unprecedented sensitivity for bosons within the 10 keV to 10 MeV mass range.

  12. Transition paths in single-molecule force spectroscopy.

    Cossio, Pilar; Hummer, Gerhard; Szabo, Attila

    2018-03-28

    In a typical single-molecule force spectroscopy experiment, the ends of the molecule of interest are connected by long polymer linkers to a pair of mesoscopic beads trapped in the focus of two laser beams. At constant force load, the total extension, i.e., the end-to-end distance of the molecule plus linkers, is measured as a function of time. In the simplest systems, the measured extension fluctuates about two values characteristic of folded and unfolded states, with occasional transitions between them. We have recently shown that molecular (un)folding rates can be recovered from such trajectories, with a small linker correction, as long as the characteristic time of the bead fluctuations is shorter than the residence time in the unfolded (folded) state. Here, we show that accurate measurements of the molecular transition path times require an even faster apparatus response. Transition paths, the trajectory segments in which the molecule (un)folds, are properly resolved only if the beads fluctuate more rapidly than the end-to-end distance of the molecule. Therefore, over a wide regime, the measured rates may be meaningful but not the transition path times. Analytic expressions for the measured mean transition path times are obtained for systems diffusing anisotropically on a two-dimensional free energy surface. The transition path times depend on the properties both of the molecule and of the pulling device.

  13. Quantifying cellular mechanics and adhesion in renal tubular injury using single cell force spectroscopy.

    Siamantouras, Eleftherios; Hills, Claire E; Squires, Paul E; Liu, Kuo-Kang

    2016-05-01

    Tubulointerstitial fibrosis represents the major underlying pathology of diabetic nephropathy where loss of cell-to-cell adhesion is a critical step. To date, research has predominantly focussed on the loss of cell surface molecular binding events that include altered protein ligation. In the current study, atomic force microscopy single cell force spectroscopy (AFM-SCFS) was used to quantify changes in cellular stiffness and cell adhesion in TGF-β1 treated kidney cells of the human proximal tubule (HK2). AFM indentation of TGF-β1 treated HK2 cells showed a significant increase (42%) in the elastic modulus (stiffness) compared to control. Fluorescence microscopy confirmed that increased cell stiffness is accompanied by reorganization of the cytoskeleton. The corresponding changes in stiffness, due to F-actin rearrangement, affected the work of detachment by changing the separation distance between two adherent cells. Overall, our novel data quantitatively demonstrate a correlation between cellular elasticity, adhesion and early morphologic/phenotypic changes associated with tubular injury. Diabetes affects many patients worldwide. One of the long term problems is diabetic nephropathy. Here, the authors utilized atomic force microscopy single cell force spectroscopy (AFM- SCFS) to study cellular stiffness and cell adhesion after TGF1 treatment in human proximal tubule kidney cells. The findings would help further understand the overall disease mechanism in diabetic patients. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. An open source/real-time atomic force microscope architecture to perform customizable force spectroscopy experiments.

    Materassi, Donatello; Baschieri, Paolo; Tiribilli, Bruno; Zuccheri, Giampaolo; Samorì, Bruno

    2009-08-01

    We describe the realization of an atomic force microscope architecture designed to perform customizable experiments in a flexible and automatic way. Novel technological contributions are given by the software implementation platform (RTAI-LINUX), which is free and open source, and from a functional point of view, by the implementation of hard real-time control algorithms. Some other technical solutions such as a new way to estimate the optical lever constant are described as well. The adoption of this architecture provides many degrees of freedom in the device behavior and, furthermore, allows one to obtain a flexible experimental instrument at a relatively low cost. In particular, we show how such a system has been employed to obtain measures in sophisticated single-molecule force spectroscopy experiments [Fernandez and Li, Science 303, 1674 (2004)]. Experimental results on proteins already studied using the same methodologies are provided in order to show the reliability of the measure system.

  15. Single molecule force spectroscopy at high data acquisition: A Bayesian nonparametric analysis

    Sgouralis, Ioannis; Whitmore, Miles; Lapidus, Lisa; Comstock, Matthew J.; Pressé, Steve

    2018-03-01

    Bayesian nonparametrics (BNPs) are poised to have a deep impact in the analysis of single molecule data as they provide posterior probabilities over entire models consistent with the supplied data, not just model parameters of one preferred model. Thus they provide an elegant and rigorous solution to the difficult problem encountered when selecting an appropriate candidate model. Nevertheless, BNPs' flexibility to learn models and their associated parameters from experimental data is a double-edged sword. Most importantly, BNPs are prone to increasing the complexity of the estimated models due to artifactual features present in time traces. Thus, because of experimental challenges unique to single molecule methods, naive application of available BNP tools is not possible. Here we consider traces with time correlations and, as a specific example, we deal with force spectroscopy traces collected at high acquisition rates. While high acquisition rates are required in order to capture dwells in short-lived molecular states, in this setup, a slow response of the optical trap instrumentation (i.e., trapped beads, ambient fluid, and tethering handles) distorts the molecular signals introducing time correlations into the data that may be misinterpreted as true states by naive BNPs. Our adaptation of BNP tools explicitly takes into consideration these response dynamics, in addition to drift and noise, and makes unsupervised time series analysis of correlated single molecule force spectroscopy measurements possible, even at acquisition rates similar to or below the trap's response times.

  16. Kinoform optics applied to X-ray photon correlation spectroscopy.

    Sandy, A R; Narayanan, S; Sprung, M; Su, J-D; Evans-Lutterodt, K; Isakovic, A F; Stein, A

    2010-05-01

    Moderate-demagnification higher-order silicon kinoform focusing lenses have been fabricated to facilitate small-angle X-ray photon correlation spectroscopy (XPCS) experiments. The geometric properties of such lenses, their focusing performance and their applicability for XPCS measurements are described. It is concluded that one-dimensional vertical X-ray focusing via silicon kinoform lenses significantly increases the usable coherent flux from third-generation storage-ring light sources for small-angle XPCS experiments.

  17. Two-dimensional correlation spectroscopy in polymer study

    Park, Yeonju; Noda, Isao; Jung, Young Mee

    2015-01-01

    This review outlines the recent works of two-dimensional correlation spectroscopy (2DCOS) in polymer study. 2DCOS is a powerful technique applicable to the in-depth analysis of various spectral data of polymers obtained under some type of perturbation. The powerful utility of 2DCOS combined with various analytical techniques in polymer studies and noteworthy developments of 2DCOS used in this field are also highlighted. PMID:25815286

  18. Combination of laser correlation and dielectric spectroscopy in albumin investigations

    Nepomnyashchaya, E; Cheremiskina, A; Velichko, E; Aksenov, E; Bogomaz, T

    2015-01-01

    Joint use of laser correlation and dielectric spectroscopies for studies of biomolecular properties of albumin in water solution is considered. The conditions and parameters of the experiments are discussed. Similar behaviours of albumin molecular sizes and maximum frequency of peak of dielectric dissipation factor with increasing acidity were revealed. Using the suggested approach, biomolecular aggregation dynamics and changes in electrophysical properties on transition from one molecular structure to another may be investigated. (paper)

  19. Recent mathematical developments in 2D correlation spectroscopy

    Noda, I.

    2000-03-01

    Recent mathematical developments in the field of 2D correlation spectroscopy, especially those related to the statistical theory, are reported. The notion of correlation phase angle is introduced. The significance of correlation phase angle between dynamic fluctuations of signals measured at two different spectral variables may be linked to more commonly known statistical concepts, such as coherence and correlation coefficient. This treatment provides the direct mathematical connection between the synchronous 2D correlation spectrum with a continuous form of the variance-covariance matrix. Moreover, it gives the background for the formal definition of the disrelation spectrum, which may be used as a heuristic substitution for the asynchronous 2D spectrum. The 2D correlation intensity may be separated into two independent factors representing the normalized extent of signal fluctuation coherence (i.e., correlation coefficient) and the magnitude of spectral intensity changes (i.e., variance). Such separation offers a convenient way to artificially enhance the discriminating power of 2D correlation spectra.

  20. Going Vertical To Improve the Accuracy of Atomic Force Microscopy Based Single-Molecule Force Spectroscopy.

    Walder, Robert; Van Patten, William J; Adhikari, Ayush; Perkins, Thomas T

    2018-01-23

    Single-molecule force spectroscopy (SMFS) is a powerful technique to characterize the energy landscape of individual proteins, the mechanical properties of nucleic acids, and the strength of receptor-ligand interactions. Atomic force microscopy (AFM)-based SMFS benefits from ongoing progress in improving the precision and stability of cantilevers and the AFM itself. Underappreciated is that the accuracy of such AFM studies remains hindered by inadvertently stretching molecules at an angle while measuring only the vertical component of the force and extension, degrading both measurements. This inaccuracy is particularly problematic in AFM studies using double-stranded DNA and RNA due to their large persistence length (p ≈ 50 nm), often limiting such studies to other SMFS platforms (e.g., custom-built optical and magnetic tweezers). Here, we developed an automated algorithm that aligns the AFM tip above the DNA's attachment point to a coverslip. Importantly, this algorithm was performed at low force (10-20 pN) and relatively fast (15-25 s), preserving the connection between the tip and the target molecule. Our data revealed large uncorrected lateral offsets for 100 and 650 nm DNA molecules [24 ± 18 nm (mean ± standard deviation) and 180 ± 110 nm, respectively]. Correcting this offset yielded a 3-fold improvement in accuracy and precision when characterizing DNA's overstretching transition. We also demonstrated high throughput by acquiring 88 geometrically corrected force-extension curves of a single individual 100 nm DNA molecule in ∼40 min and versatility by aligning polyprotein- and PEG-based protein-ligand assays. Importantly, our software-based algorithm was implemented on a commercial AFM, so it can be broadly adopted. More generally, this work illustrates how to enhance AFM-based SMFS by developing more sophisticated data-acquisition protocols.

  1. Investigation of polyelectrolyte desorption by single molecule force spectroscopy

    Friedsam, C; Seitz, M; Gaub, H E

    2004-01-01

    Single molecule force spectroscopy has evolved into a powerful method for the investigation of intra- and intermolecular interactions at the level of individual molecules. Many examples, including the investigation of the dynamic properties of complex biological systems as well as the properties of covalent bonds or intermolecular transitions within individual polymers, are reported in the literature. The technique has recently been extended to the systematic investigation of desorption processes of individual polyelectrolyte molecules adsorbed on generic surfaces. The stable covalent attachment of polyelectrolyte molecules to the AFM-tip provides the possibility of performing long-term measurements with the same set of molecules and therefore allows the in situ observation of the impact of environmental changes on the adsorption behaviour of individual molecules. Different types of interactions, e.g. electrostatic or hydrophobic interactions, that determine the adsorption process could be identified and characterized. The experiments provided valuable details that help to understand the nature and the properties of non-covalent interactions, which is helpful with regard to biological systems as well as for technical applications. Apart from this, desorption experiments can be utilized to characterize the properties of surfaces or polymer coatings. Therefore they represent a versatile tool that can be further developed in terms of various aspects

  2. Oxygen measurement by multimode diode lasers employing gas correlation spectroscopy.

    Lou, Xiutao; Somesfalean, Gabriel; Chen, Bin; Zhang, Zhiguo

    2009-02-10

    Multimode diode laser (MDL)-based correlation spectroscopy (COSPEC) was used to measure oxygen in ambient air, thereby employing a diode laser (DL) having an emission spectrum that overlaps the oxygen absorption lines of the A band. A sensitivity of 700 ppm m was achieved with good accuracy (2%) and linearity (R(2)=0.999). For comparison, measurements of ambient oxygen were also performed by tunable DL absorption spectroscopy (TDLAS) technique employing a vertical cavity surface emitting laser. We demonstrate that, despite slightly degraded sensitivity, the MDL-based COSPEC-based oxygen sensor has the advantages of high stability, low cost, ease-of-use, and relaxed requirements in component selection and instrument buildup compared with the TDLAS-based instrument.

  3. Resonant Optical Gradient Force Interaction for Nano-Imaging and-Spectroscopy

    2016-07-19

    New J. Phys. 18 (2016) 053042 doi:10.1088/1367-2630/18/5/053042 PAPER Resonant optical gradient force interaction for nano-imaging and -spectroscopy...HonghuaUYang andMarkus BRaschke Department of Physics , Department of Chemistry, and JILA,University of Colorado, Boulder, CO80309,USA E-mail...honghua.yang@colorado.edu andmarkus.raschke@colorado.edu Keywords:nano spectroscopy, optical force, near-field optics Abstract The optical gradient force

  4. Detection of elemental mercury by multimode diode laser correlation spectroscopy.

    Lou, Xiutao; Somesfalean, Gabriel; Svanberg, Sune; Zhang, Zhiguo; Wu, Shaohua

    2012-02-27

    We demonstrate a method for elemental mercury detection based on correlation spectroscopy employing UV laser radiation generated by sum-frequency mixing of two visible multimode diode lasers. Resonance matching of the multimode UV laser is achieved in a wide wavelength range and with good tolerance for various operating conditions. Large mode-hops provide an off-resonance baseline, eliminating interferences from other gas species with broadband absorption. A sensitivity of 1 μg/m3 is obtained for a 1-m path length and 30-s integration time. The performance of the system shows promise for mercury monitoring in industrial applications.

  5. Electron correlation effects in XUV photoabsorption spectroscopy of atoms

    Codling, K.

    1976-01-01

    Reference is made to sophisticated experiments involving the measurement of the angular distribution of photo-ejected electrons, coincidence electrons and ion spectroscopy, which can only be interpreted in terms of electron correlation effects. After an introductory review of previous work, the lectures fall under the following headings: experimental procedures (light sources, monochromators, absorption cells, limitations on the simple photoasbsorption experiment, and complementary techniques); experimental results (discrete states in the continuum, gross features in the photoionisation continuum (rare gases, alkalis, alkaline earths, rare earths, transition elements)). (U.K.)

  6. Force induced unzipping of DNA with long range correlated noise

    Lam, Pui-Man; Zhen, Yi

    2011-01-01

    We derive and solve a Fokker–Planck equation for the stationary distribution of the free energy, in a model of unzipping of double-stranded DNA under external force. The autocorrelation function of the random DNA sequence can be of a general form, including long range correlations. In the case of Ornstein–Uhlenbeck noise, characterized by a finite correlation length, our result reduces to the exact result of Allahverdyan et al, with the average number of unzipped base pairs going as (X) ∼ 1/f 2 in the white noise limit, where f is the deviation from the critical force. In the case of long range correlated noise, where the integrated autocorrelation is divergent, we find that (X) is finite at f = 0, with its value decreasing as the correlations become of longer range. This shows that long range correlations actually stabilize the DNA sequence against unzipping. Our result is also in agreement with the findings of Allahverdyan et al obtained using numerical generation of the long range correlated noise

  7. Comparative analysis of heat transfer correlations for forced convection boiling

    Guglielmini, G.; Nannei, E.; Pisoni, C.

    1978-01-01

    A critical survey was conducted of the most relevant correlations of boiling heat transfer in forced convection flow. Most of the investigations carried out on partial nucleate boiling and fully developed nucleate boiling have led to the formulation of correlations that are not able to cover a wide range of operating conditions, due to the empirical approach of the problem. A comparative analysis is therefore required in order to delineate the relative accuracy of the proposed correlations, on the basis of the experimental data presently available. The survey performed allows the evaluation of the accuracy of the different calculating procedure; the results obtained, moreover, indicate the most reliable heat transfer correlations for the different operating conditions investigated. This survey was developed for five pressure range (up to 180bar) and for both saturation and subcooled boiling condition

  8. Single-molecule force-conductance spectroscopy of hydrogen-bonded complexes

    Pirrotta, Alessandro; De Vico, Luca; Solomon, Gemma C.; Franco, Ignacio

    2017-03-01

    The emerging ability to study physical properties at the single-molecule limit highlights the disparity between what is observable in an ensemble of molecules and the heterogeneous contributions of its constituent parts. A particularly convenient platform for single-molecule studies are molecular junctions where forces and voltages can be applied to individual molecules, giving access to a series of electromechanical observables that can form the basis of highly discriminating multidimensional single-molecule spectroscopies. Here, we computationally examine the ability of force and conductance to inform about molecular recognition events at the single-molecule limit. For this, we consider the force-conductance characteristics of a prototypical class of hydrogen bonded bimolecular complexes sandwiched between gold electrodes. The complexes consist of derivatives of a barbituric acid and a Hamilton receptor that can form up to six simultaneous hydrogen bonds. The simulations combine classical molecular dynamics of the mechanical deformation of the junction with non-equilibrium Green's function computations of the electronic transport. As shown, in these complexes hydrogen bonds mediate transport either by directly participating as a possible transport pathway or by stabilizing molecular conformations with enhanced conductance properties. Further, we observe that force-conductance correlations can be very sensitive to small changes in the chemical structure of the complexes and provide detailed information about the behavior of single molecules that cannot be gleaned from either measurement alone. In fact, there are regions during the elongation that are only mechanically active, others that are only conductance active, and regions where both force and conductance changes as the complex is mechanically manipulated. The implication is that force and conductance provide complementary information about the evolution of molecules in junctions that can be used to

  9. Scanning, non-contact, hybrid broadband diffuse optical spectroscopy and diffuse correlation spectroscopy system.

    Johansson, Johannes D; Mireles, Miguel; Morales-Dalmau, Jordi; Farzam, Parisa; Martínez-Lozano, Mar; Casanovas, Oriol; Durduran, Turgut

    2016-02-01

    A scanning system for small animal imaging using non-contact, hybrid broadband diffuse optical spectroscopy (ncDOS) and diffuse correlation spectroscopy (ncDCS) is presented. The ncDOS uses a two-dimensional spectrophotometer retrieving broadband (610-900 nm) spectral information from up to fifty-seven source-detector distances between 2 and 5 mm. The ncDCS data is simultaneously acquired from four source-detector pairs. The sample is scanned in two dimensions while tracking variations in height. The system has been validated with liquid phantoms, demonstrated in vivo on a human fingertip during an arm cuff occlusion and on a group of mice with xenoimplanted renal cell carcinoma.

  10. Two-dimensional fluorescence lifetime correlation spectroscopy. 2. Application.

    Ishii, Kunihiko; Tahara, Tahei

    2013-10-03

    In the preceding article, we introduced the theoretical framework of two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS). In this article, we report the experimental implementation of 2D FLCS. In this method, two-dimensional emission-delay correlation maps are constructed from the photon data obtained with the time-correlated single photon counting (TCSPC), and then they are converted to 2D lifetime correlation maps by the inverse Laplace transform. We develop a numerical method to realize reliable transformation, employing the maximum entropy method (MEM). We apply the developed actual 2D FLCS to two real systems, a dye mixture and a DNA hairpin. For the dye mixture, we show that 2D FLCS is experimentally feasible and that it can identify different species in an inhomogeneous sample without any prior knowledge. The application to the DNA hairpin demonstrates that 2D FLCS can disclose microsecond spontaneous dynamics of biological molecules in a visually comprehensible manner, through identifying species as unique lifetime distributions. A FRET pair is attached to the both ends of the DNA hairpin, and the different structures of the DNA hairpin are distinguished as different fluorescence lifetimes in 2D FLCS. By constructing the 2D correlation maps of the fluorescence lifetime of the FRET donor, the equilibrium dynamics between the open and the closed forms of the DNA hairpin is clearly observed as the appearance of the cross peaks between the corresponding fluorescence lifetimes. This equilibrium dynamics of the DNA hairpin is clearly separated from the acceptor-missing DNA that appears as an isolated diagonal peak in the 2D maps. The present study clearly shows that newly developed 2D FLCS can disclose spontaneous structural dynamics of biological molecules with microsecond time resolution.

  11. Transfer of spectral weight in spectroscopies of correlated electron systems

    Rozenberg, M.J.; Kotliar, G.; Kajueter, H.

    1996-01-01

    We study the transfer of spectral weight in the photoemission and optical spectra of strongly correlated electron systems. Within the local impurity self-consistent approximation, that becomes exact in the limit of large lattice coordination, we consider and compare two models of correlated electrons, the Hubbard model and the periodic Anderson model. The results are discussed in regard to recent experiments. In the Hubbard model, we predict an anomalous enhancement optical spectral weight as a function of temperature in the correlated metallic state which is in qualitative agreement with optical measurements in V 2 O 3 . We argue that anomalies observed in the spectroscopy of the metal are connected to the proximity to a crossover region in the phase diagram of the model. In the insulating phase, we obtain excellent agreement with the experimental data, and present a detailed discussion on the role of magnetic frustration by studying the k-resolved single-particle spectra. The results for the periodic Anderson model are discussed in connection to recent experimental data of the Kondo insulators Ce 3 Bi 4 Pt 3 and FeSi. The model can successfully explain the thermal filling of the optical gap and the corresponding changes in the photoemission density of states. The temperature dependence of the optical sum rule is obtained, and its relevance to the interpretation of the experimental data discussed. Finally, we argue that the large scattering rate measured in Kondo insulators cannot be described by the periodic Anderson model. copyright 1996 The American Physical Society

  12. LASER CORRELATION SPECTROSCOPY (LCS AND ITS CLINICAL PERSPECTIVES IN OPHTHALMOLOGY

    Karganov Mikhail

    2015-12-01

    Full Text Available The method of laser correlation spectroscopy (LCS is based on the analysis of the spectrum of quasielastic light scatter during coherent monochromatic laser irradiation of micro-particles in biological fluids (blood serum, urine, oropharyngeal washout fluid, tear fluid etc.. Spectrum provides information on dynamic processes in the analyzed system: translation motion of scattering particles and their orientation and conformation dynamics. Special procedures of cluster analysis make it possible to find out to which linkage group a particular spectrum belongs. LCS allows evaluation of sub-fractional composition of biological fluids in a wide range of molecular sizes (from 1 to 10,000 nm, which determines principal novelty of this approach in ophthalmology.

  13. Split and delay photon correlation spectroscopy with a visible light

    Rasch, Marten

    2016-04-01

    The development and performance of a setup constructed with the aim for the split pulse photon correlation spectroscopy is presented in this thesis. The double pulse time structure is accomplished with help of an Acusto-Optic Modulator (AOM) crystal, which mimics the splitting and delaying of photon pulses. The setup provides double pulses and allows to control the pulse width and delay and to synchronize them into one camera exposure window. The performance of the setup was successfully verified in a proof of principle experiment with a model system of polystyrene particles following Brownian motion. The measured radius of particles obtained with from the split pulse experiment (R h =(2.567±0.097) μm) is in agreement with the particle size provided by the manufacturer (R=(2.26±0.08) μm). The achieved results show higher statistics compared to a standard Dynamic Light Scattering (DLS) measurement.

  14. Dynamic force spectroscopy of oppositely charged polyelectrolyte brushes

    Spruijt, E.; Cohen Stuart, M.A.; Gucht, van der J.

    2010-01-01

    Ion pairing is the main driving force in the formation of polyelectrolyte complexes, which find widespread use in micellar assemblies, drug carriers, and coatings. In this paper we examine the actual ion pairing forces in a polyelectrolyte complex between two oppositely charged polyelectrolyte

  15. Resolving Fast, Confined Diffusion in Bacteria with Image Correlation Spectroscopy.

    Rowland, David J; Tuson, Hannah H; Biteen, Julie S

    2016-05-24

    By following single fluorescent molecules in a microscope, single-particle tracking (SPT) can measure diffusion and binding on the nanometer and millisecond scales. Still, although SPT can at its limits characterize the fastest biomolecules as they interact with subcellular environments, this measurement may require advanced illumination techniques such as stroboscopic illumination. Here, we address the challenge of measuring fast subcellular motion by instead analyzing single-molecule data with spatiotemporal image correlation spectroscopy (STICS) with a focus on measurements of confined motion. Our SPT and STICS analysis of simulations of the fast diffusion of confined molecules shows that image blur affects both STICS and SPT, and we find biased diffusion rate measurements for STICS analysis in the limits of fast diffusion and tight confinement due to fitting STICS correlation functions to a Gaussian approximation. However, we determine that with STICS, it is possible to correctly interpret the motion that blurs single-molecule images without advanced illumination techniques or fast cameras. In particular, we present a method to overcome the bias due to image blur by properly estimating the width of the correlation function by directly calculating the correlation function variance instead of using the typical Gaussian fitting procedure. Our simulation results are validated by applying the STICS method to experimental measurements of fast, confined motion: we measure the diffusion of cytosolic mMaple3 in living Escherichia coli cells at 25 frames/s under continuous illumination to illustrate the utility of STICS in an experimental parameter regime for which in-frame motion prevents SPT and tight confinement of fast diffusion precludes stroboscopic illumination. Overall, our application of STICS to freely diffusing cytosolic protein in small cells extends the utility of single-molecule experiments to the regime of fast confined diffusion without requiring advanced

  16. Establishing the diffuse correlation spectroscopy signal relationship with blood flow.

    Boas, David A; Sakadžić, Sava; Selb, Juliette; Farzam, Parisa; Franceschini, Maria Angela; Carp, Stefan A

    2016-07-01

    Diffuse correlation spectroscopy (DCS) measurements of blood flow rely on the sensitivity of the temporal autocorrelation function of diffusively scattered light to red blood cell (RBC) mean square displacement (MSD). For RBCs flowing with convective velocity [Formula: see text], the autocorrelation is expected to decay exponentially with [Formula: see text], where [Formula: see text] is the delay time. RBCs also experience shear-induced diffusion with a diffusion coefficient [Formula: see text] and an MSD of [Formula: see text]. Surprisingly, experimental data primarily reflect diffusive behavior. To provide quantitative estimates of the relative contributions of convective and diffusive movements, we performed Monte Carlo simulations of light scattering through tissue of varying vessel densities. We assumed laminar vessel flow profiles and accounted for shear-induced diffusion effects. In agreement with experimental data, we found that diffusive motion dominates the correlation decay for typical DCS measurement parameters. Furthermore, our model offers a quantitative relationship between the RBC diffusion coefficient and absolute tissue blood flow. We thus offer, for the first time, theoretical support for the empirically accepted ability of the DCS blood flow index ([Formula: see text]) to quantify tissue perfusion. We find [Formula: see text] to be linearly proportional to blood flow, but with a proportionality modulated by the hemoglobin concentration and the average blood vessel diameter.

  17. Multiplexed single-molecule force spectroscopy using a centrifuge.

    Yang, Darren; Ward, Andrew; Halvorsen, Ken; Wong, Wesley P

    2016-03-17

    We present a miniature centrifuge force microscope (CFM) that repurposes a benchtop centrifuge for high-throughput single-molecule experiments with high-resolution particle tracking, a large force range, temperature control and simple push-button operation. Incorporating DNA nanoswitches to enable repeated interrogation by force of single molecular pairs, we demonstrate increased throughput, reliability and the ability to characterize population heterogeneity. We perform spatiotemporally multiplexed experiments to collect 1,863 bond rupture statistics from 538 traceable molecular pairs in a single experiment, and show that 2 populations of DNA zippers can be distinguished using per-molecule statistics to reduce noise.

  18. Optimized Free Energies from Bidirectional Single-Molecule Force Spectroscopy

    Minh, David D. L.; Adib, Artur B.

    2008-05-01

    An optimized method for estimating path-ensemble averages using data from processes driven in opposite directions is presented. Based on this estimator, bidirectional expressions for reconstructing free energies and potentials of mean force from single-molecule force spectroscopy—valid for biasing potentials of arbitrary stiffness—are developed. Numerical simulations on a model potential indicate that these methods perform better than unidirectional strategies.

  19. An in vitro correlation of mechanical forces and metastatic capacity

    Indra, Indrajyoti; Undyala, Vishnu; Kandow, Casey; Thirumurthi, Umadevi; Beningo, Karen A; Dembo, Micah

    2011-01-01

    Mechanical forces have a major influence on cell migration and are predicted to significantly impact cancer metastasis, yet this idea is currently poorly defined. In this study we have asked if changes in traction stress and migratory properties correlate with the metastatic progression of tumor cells. For this purpose, four murine breast cancer cell lines derived from the same primary tumor, but possessing increasing metastatic capacity, were tested for adhesion strength, traction stress, focal adhesion organization and for differential migration rates in two-dimensional and three-dimensional environments. Using traction force microscopy (TFM), we were surprised to find an inverse relationship between traction stress and metastatic capacity, such that force production decreased as the metastatic capacity increased. Consistent with this observation, adhesion strength exhibited an identical profile to the traction data. A count of adhesions indicated a general reduction in the number as metastatic capacity increased but no difference in the maturation as determined by the ratio of nascent to mature adhesions. These changes correlated well with a reduction in active beta-1 integrin with increasing metastatic ability. Finally, in two dimensions, wound healing, migration and persistence were relatively low in the entire panel, maintaining a downward trend with increasing metastatic capacity. Why metastatic cells would migrate so poorly prompted us to ask if the loss of adhesive parameters in the most metastatic cells indicated a switch to a less adhesive mode of migration that would only be detected in a three-dimensional environment. Indeed, in three-dimensional migration assays, the most metastatic cells now showed the greatest linear speed. We conclude that traction stress, adhesion strength and rate of migration do indeed change as tumor cells progress in metastatic capacity and do so in a dimension-sensitive manner

  20. Reverse engineering of an affinity-switchable molecular interaction characterized by atomic force microscopy single-molecule force spectroscopy.

    Anselmetti, Dario; Bartels, Frank Wilco; Becker, Anke; Decker, Björn; Eckel, Rainer; McIntosh, Matthew; Mattay, Jochen; Plattner, Patrik; Ros, Robert; Schäfer, Christian; Sewald, Norbert

    2008-02-19

    Tunable and switchable interaction between molecules is a key for regulation and control of cellular processes. The translation of the underlying physicochemical principles to synthetic and switchable functional entities and molecules that can mimic the corresponding molecular functions is called reverse molecular engineering. We quantitatively investigated autoinducer-regulated DNA-protein interaction in bacterial gene regulation processes with single atomic force microscopy (AFM) molecule force spectroscopy in vitro, and developed an artificial bistable molecular host-guest system that can be controlled and regulated by external signals (UV light exposure and thermal energy). The intermolecular binding functionality (affinity) and its reproducible and reversible switching has been proven by AFM force spectroscopy at the single-molecule level. This affinity-tunable optomechanical switch will allow novel applications with respect to molecular manipulation, nanoscale rewritable molecular memories, and/or artificial ion channels, which will serve for the controlled transport and release of ions and neutral compounds in the future.

  1. Quantifying DNA melting transitions using single-molecule force spectroscopy

    Calderon, Christopher P; Chen, W-H; Harris, Nolan C; Kiang, C-H; Lin, K-J

    2009-01-01

    We stretched a DNA molecule using an atomic force microscope (AFM) and quantified the mechanical properties associated with B and S forms of double-stranded DNA (dsDNA), molten DNA, and single-stranded DNA. We also fit overdamped diffusion models to the AFM time series and used these models to extract additional kinetic information about the system. Our analysis provides additional evidence supporting the view that S-DNA is a stable intermediate encountered during dsDNA melting by mechanical force. In addition, we demonstrated that the estimated diffusion models can detect dynamical signatures of conformational degrees of freedom not directly observed in experiments.

  2. Quantifying DNA melting transitions using single-molecule force spectroscopy

    Calderon, Christopher P [Department of Computational and Applied Mathematics, Rice University, Houston, TX (United States); Chen, W-H; Harris, Nolan C; Kiang, C-H [Department of Physics and Astronomy, Rice University, Houston, TX (United States); Lin, K-J [Department of Chemistry, National Chung Hsing University, Taichung, Taiwan (China)], E-mail: chkiang@rice.edu

    2009-01-21

    We stretched a DNA molecule using an atomic force microscope (AFM) and quantified the mechanical properties associated with B and S forms of double-stranded DNA (dsDNA), molten DNA, and single-stranded DNA. We also fit overdamped diffusion models to the AFM time series and used these models to extract additional kinetic information about the system. Our analysis provides additional evidence supporting the view that S-DNA is a stable intermediate encountered during dsDNA melting by mechanical force. In addition, we demonstrated that the estimated diffusion models can detect dynamical signatures of conformational degrees of freedom not directly observed in experiments.

  3. Reassessment of forced convection heat transfer correlations for refrigerant-12

    Celata, G.P.; Cuomo, M.; D'Annibale, F.; Farello, G.E.; Setaro, T.

    1986-01-01

    In the frame of a Refrigerant-12 experiment on postulated accidental transients in Pressurized Water Reactors under way at Heat Transfer Laboratory (ENEA Casaccia Research Center), an assessment of the main correlation available in scientific literature, for the different heat transfer regions encountered when a liquid is boiled in a confined heated channel, has been performed. Considering a vertical tube uniformly heated over its length with CHF at the exit, the following heat transfer regimes may be individuated: convective heat transfer to liquid, subcooled boiling, saturated nucleate boiling, forced convective heat transfer through liquid film (annular flow regime) and thermal crisis. From the comparison of computed values with an original ENEA dataset, the best correlations in predicting Refrigerant-12 data have been individuated. In a few cases, though preserving the original structure of the correlations, mainly developed for water, it was necessary to adjust some coefficients by means of best-fit procedures through our experimental data. The work has been performed in the frame of the ENEA Thermal Reactor Department Safety Research Project

  4. Dissipation and oscillatory solvation forces in confined liquids studied by small amplitude atomic force spectroscopy

    de Beer, Sissi; van den Ende, Henricus T.M.; Mugele, Friedrich

    2010-01-01

    We determine conservative and dissipative tip–sample interaction forces from the amplitude and phase response of acoustically driven atomic force microscope (AFM) cantilevers using a non-polar model fluid (octamethylcyclotetrasiloxane, which displays strong molecular layering) and atomically flat

  5. Inference of protein diffusion probed via fluorescence correlation spectroscopy

    Tsekouras, Konstantinos

    2015-03-01

    Fluctuations are an inherent part of single molecule or few particle biophysical data sets. Traditionally, ``noise'' fluctuations have been viewed as a nuisance, to be eliminated or minimized. Here we look on how statistical inference methods - that take explicit advantage of fluctuations - have allowed us to draw an unexpected picture of single molecule diffusional dynamics. Our focus is on the diffusion of proteins probed using fluorescence correlation spectroscopy (FCS). First, we discuss how - in collaboration with the Bustamante and Marqusee labs at UC Berkeley - we determined using FCS data that individual enzymes are perturbed by self-generated catalytic heat (Riedel et al, Nature, 2014). Using the tools of inference, we found how distributions of enzyme diffusion coefficients shift in the presence of substrate revealing that enzymes performing highly exothermic reactions dissipate heat by transiently accelerating their center of mass following a catalytic reaction. Next, when molecules diffuse in the cell nucleus they often appear to diffuse anomalously. We analyze FCS data - in collaboration with Rich Day at the IU Med School - to propose a simple model for transcription factor binding-unbinding in the nucleus to show that it may give rise to apparent anomalous diffusion. Here inference methods extract entire binding affinity distributions for the diffusing transcription factors, allowing us to precisely characterize their interactions with different components of the nuclear environment. From this analysis, we draw key mechanistic insight that goes beyond what is possible by simply fitting data to ``anomalous diffusion'' models.

  6. Theory of L -edge spectroscopy of strongly correlated systems

    Lüder, Johann; Schött, Johan; Brena, Barbara; Haverkort, Maurits W.; Thunström, Patrik; Eriksson, Olle; Sanyal, Biplab; Di Marco, Igor; Kvashnin, Yaroslav O.

    2017-12-01

    X-ray absorption spectroscopy measured at the L edge of transition metals (TMs) is a powerful element-selective tool providing direct information about the correlation effects in the 3 d states. The theoretical modeling of the 2 p →3 d excitation processes remains to be challenging for contemporary ab initio electronic structure techniques, due to strong core-hole and multiplet effects influencing the spectra. In this work, we present a realization of the method combining the density-functional theory with multiplet ligand field theory, proposed in Haverkort et al. [Phys. Rev. B 85, 165113 (2012), 10.1103/PhysRevB.85.165113]. In this approach, a single-impurity Anderson model (SIAM) is constructed, with almost all parameters obtained from first principles, and then solved to obtain the spectra. In our implementation, we adopt the language of the dynamical mean-field theory and utilize the local density of states and the hybridization function, projected onto TM 3 d states, in order to construct the SIAM. The developed computational scheme is applied to calculate the L -edge spectra for several TM monoxides. A very good agreement between the theory and experiment is found for all studied systems. The effect of core-hole relaxation, hybridization discretization, possible extensions of the method as well as its limitations are discussed.

  7. Progress in the Correlative Atomic Force Microscopy and Optical Microscopy

    Lulu Zhou

    2017-04-01

    Full Text Available Atomic force microscopy (AFM has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

  8. Assessment of sacrococcygeal pressure ulcers using diffuse correlation spectroscopy

    Diaz, David; Lafontant, Alec; Neidrauer, Michael; Weingarten, Michael S.; DiMaria-Ghalili, Rose Ann; Fried, Guy W.; Rece, Julianne; Lewin, Peter A.; Zubkov, Leonid

    2016-03-01

    Microcirculation is essential for proper supply of oxygen and nutritive substances to the biological tissue and the removal of waste products of metabolism. The determination of microcirculatory blood flow (mBF) is therefore of substantial interest to clinicians for assessing tissue health; particularly in pressure ulceration and suspected deep tissue injury. The goal of this pilot clinical study was to assess deep-tissue pressure ulceration by non-invasively measuring mBF using Diffuse Correlation Spectroscopy (DCS). DCS provides information about the flow of red blood cells in the capillary network by measuring the temporal autocorrelation function of scattering light intensity. A novel optical probe was developed in order to obtain measurements under the load of the subject's body as pressure is applied (ischemia) and then released (reperfusion) on sacrococcygeal tissue in a hospital bed. Prior to loading measurements, baseline readings of the sacral region were obtained by measuring the subjects in a side-lying position. DCS measurements from the sacral region of twenty healthy volunteers have been compared to those of two patients who initially had similar non-blanchable redness. The temporal autocorrelation function of scattering light intensity of the patient whose redness later disappeared was similar to that of the average healthy subject. The second patient, whose redness developed into an advanced pressure ulcer two weeks later, had a substantial decrease in blood flow while under the loading position compared to healthy subjects. Preliminary results suggest the developed system may potentially predict whether non-blanchable redness will manifest itself as advanced ulceration or dissipate over time.

  9. A wind loading correlation for an isolated square heliostat, part 1: lift and drag forces

    Roos, TH

    2012-05-01

    Full Text Available dataset to high accuracy. Correlations for the lift and drag forces are presented. A companion paper presents correlations for the side forces and correlations for moments about the three principal axes, and discusses the behavior of the correlations....

  10. Non-equilibrium umbrella sampling applied to force spectroscopy of soft matter.

    Gao, Y X; Wang, G M; Williams, D R M; Williams, Stephen R; Evans, Denis J; Sevick, E M

    2012-02-07

    Physical systems often respond on a timescale which is longer than that of the measurement. This is particularly true in soft matter where direct experimental measurement, for example in force spectroscopy, drives the soft system out of equilibrium and provides a non-equilibrium measure. Here we demonstrate experimentally for the first time that equilibrium physical quantities (such as the mean square displacement) can be obtained from non-equilibrium measurements via umbrella sampling. Our model experimental system is a bead fluctuating in a time-varying optical trap. We also show this for simulated force spectroscopy on a complex soft molecule--a piston-rotaxane.

  11. Atomic force microscope-assisted scanning tunneling spectroscopy under ambient conditions.

    Vakhshouri, Amin; Hashimoto, Katsushi; Hirayama, Yoshiro

    2014-12-01

    We have developed a method of atomic force microscopy (AFM)-assisted scanning tunneling spectroscopy (STS) under ambient conditions. An AFM function is used for rapid access to a selected position prior to performing STS. The AFM feedback is further used to suppress vertical thermal drift of the tip-sample distance during spectroscopy, enabling flexible and stable spectroscopy measurements at room temperature. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Antibody-Unfolding and Metastable-State Binding in Force Spectroscopy and Recognition Imaging

    Kaur, Parminder; Qiang-Fu; Fuhrmann, Alexander; Ros, Robert; Kutner, Linda Obenauer; Schneeweis, Lumelle A.; Navoa, Ryman; Steger, Kirby; Xie, Lei; Yonan, Christopher; Abraham, Ralph; Grace, Michael J.; Lindsay, Stuart

    2011-01-01

    Force spectroscopy and recognition imaging are important techniques for characterizing and mapping molecular interactions. In both cases, an antibody is pulled away from its target in times that are much less than the normal residence time of the antibody on its target. The distribution of pulling lengths in force spectroscopy shows the development of additional peaks at high loading rates, indicating that part of the antibody frequently unfolds. This propensity to unfold is reversible, indicating that exposure to high loading rates induces a structural transition to a metastable state. Weakened interactions of the antibody in this metastable state could account for reduced specificity in recognition imaging where the loading rates are always high. The much weaker interaction between the partially unfolded antibody and target, while still specific (as shown by control experiments), results in unbinding on millisecond timescales, giving rise to rapid switching noise in the recognition images. At the lower loading rates used in force spectroscopy, we still find discrepancies between the binding kinetics determined by force spectroscopy and those determined by surface plasmon resonance—possibly a consequence of the short tethers used in recognition imaging. Recognition imaging is nonetheless a powerful tool for interpreting complex atomic force microscopy images, so long as specificity is calibrated in situ, and not inferred from equilibrium binding kinetics. PMID:21190677

  13. Atomic force and shear force based tip-enhanced Raman spectroscopy and imaging

    Kharintsev, S.S.; Hoffmann, G.G.; Dorozhkin, P.S.; With, de G.; Loos, J.

    2007-01-01

    Underlying near-field optibal effects on the nanoscale have stimulated the development of apertureless vibrational spectroscopy and imaging with ultrahigh spatial resolution. We demonstrate tip-enhanced Raman spectra of single-walled carbon nanotubes (SWCNTs), recorded with a scanning near-field

  14. Understanding image contrast formation in TiO.sub.2./sub. with force spectroscopy

    Yurtsever, A.; Fernandez-Torre, D.; González, C.; Jelínek, Pavel; Pou, P.; Sugimoto, Y.; Abe, M.; Pérez, R.; Morita, S.

    2012-01-01

    Roč. 85, č. 12 (2012), "125416-1"-"125416-9" ISSN 1098-0121 R&D Projects: GA MŠk(CZ) ME10076 Institutional research plan: CEZ:AV0Z10100521 Keywords : DFT * AFM * force spectroscopy * atomic resolution Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 3.767, year: 2012

  15. Investigation of the heparin-thrombin interaction by dynamic force spectroscopy.

    Wang, Congzhou; Jin, Yingzi; Desai, Umesh R; Yadavalli, Vamsi K

    2015-06-01

    The interaction between heparin and thrombin is a vital step in the blood (anti)coagulation process. Unraveling the molecular basis of the interactions is therefore extremely important in understanding the mechanisms of this complex biological process. In this study, we use a combination of an efficient thiolation chemistry of heparin, a self-assembled monolayer-based single molecule platform, and a dynamic force spectroscopy to provide new insights into the heparin-thrombin interaction from an energy viewpoint at the molecular scale. Well-separated single molecules of heparin covalently attached to mixed self-assembled monolayers are demonstrated, whereby interaction forces with thrombin can be measured via atomic force microscopy-based spectroscopy. Further these interactions are studied at different loading rates and salt concentrations to directly obtain kinetic parameters. An increase in the loading rate shows a higher interaction force between the heparin and thrombin, which can be directly linked to the kinetic dissociation rate constant (koff). The stability of the heparin/thrombin complex decreased with increasing NaCl concentration such that the off-rate was found to be driven primarily by non-ionic forces. These results contribute to understanding the role of specific and nonspecific forces that drive heparin-thrombin interactions under applied force or flow conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Numerical analysis of dynamic force spectroscopy using the torsional harmonic cantilever

    Solares, Santiago D; Hoelscher, Hendrik

    2010-01-01

    A spectral analysis method has been recently introduced by Stark et al (2002 Proc. Natl Acad. Sci. USA 99 8473-8) and implemented by Sahin et al (2007 Nat. Nanotechnol. 2 507-14) using a T-shaped cantilever design, the torsional harmonic cantilever (THC), which is capable of performing simultaneous tapping-mode atomic force microscopy imaging and force spectroscopy. Here we report on numerical simulations of the THC system using a simple dual-mass flexural-torsional model, which is applied in combination with Fourier data processing software to illustrate the spectroscopy process for quality factors corresponding to liquid, air and vacuum environments. We also illustrate the acquisition of enhanced topographical images and deformed surface contours under the application of uniform forces, and compare the results to those obtained with a previously reported linear dual-spring-mass model.

  17. Atomic force microscopy imaging and single molecule recognition force spectroscopy of coat proteins on the surface of Bacillus subtilis spore.

    Tang, Jilin; Krajcikova, Daniela; Zhu, Rong; Ebner, Andreas; Cutting, Simon; Gruber, Hermann J; Barak, Imrich; Hinterdorfer, Peter

    2007-01-01

    Coat assembly in Bacillus subtilis serves as a tractable model for the study of the self-assembly process of biological structures and has a significant potential for use in nano-biotechnological applications. In the present study, the morphology of B. subtilis spores was investigated by magnetically driven dynamic force microscopy (MAC mode atomic force microscopy) under physiological conditions. B. subtilis spores appeared as prolate structures, with a length of 0.6-3 microm and a width of about 0.5-2 microm. The spore surface was mainly covered with bump-like structures with diameters ranging from 8 to 70 nm. Besides topographical explorations, single molecule recognition force spectroscopy (SMRFS) was used to characterize the spore coat protein CotA. This protein was specifically recognized by a polyclonal antibody directed against CotA (anti-CotA), the antibody being covalently tethered to the AFM tip via a polyethylene glycol linker. The unbinding force between CotA and anti-CotA was determined as 55 +/- 2 pN. From the high-binding probability of more than 20% in force-distance cycles it is concluded that CotA locates in the outer surface of B. subtilis spores. Copyright (c) 2007 John Wiley & Sons, Ltd.

  18. Nanopuller-open data acquisition platform for AFM force spectroscopy experiments

    Pawlak, Konrad; Strzelecki, Janusz

    2016-01-01

    Atomic Force Microscope (AFM) is a widely used tool in force spectroscopy studies. Presently, this instrument is accessible from numerous vendors, albeit commercial solutions are expensive and almost always hardware and software closed. Approaches for open setups were published, as with modern low cost and readily available piezoelectric actuators, data acquisition interfaces and optoelectronic components building such force spectroscopy AFM is relatively easy. However, suitable software to control such laboratory made instrument was not released. Developing it in the lab requires significant time and effort. Our Nanopuller software described in this paper is intended to eliminate this obstacle. With only minimum adjustments this program can be used to control and acquire data with any suitable National Instruments universal digital/analog interface and piezoelectric actuator analog controller, giving significant freedom and flexibility in designing force spectroscopy experiment. Since the full code, written in a graphical LabVIEW environment is available, our Nanopuller can be easily customized. In this paper we describe the program and test its performance in controlling different setups. Successful and accurate force curve acquisition for standard samples (single molecules of I27O reference titin polyprotein and DNA as well as red blood cells) is shown. - Highlights: • We created open data acquisition software for performing Atomic Force Microscopy force measurements with custom laboratory made setups. • The software allows large flexibility in atomic force microscope design with minimum adjustment necessary. • The software is written in LabVIEW, allowing easy customization. • We successfully tested the program on two different hardware configurations by stretching single macromolecules and indenting cells.

  19. Nanopuller-open data acquisition platform for AFM force spectroscopy experiments

    Pawlak, Konrad; Strzelecki, Janusz

    2016-05-15

    Atomic Force Microscope (AFM) is a widely used tool in force spectroscopy studies. Presently, this instrument is accessible from numerous vendors, albeit commercial solutions are expensive and almost always hardware and software closed. Approaches for open setups were published, as with modern low cost and readily available piezoelectric actuators, data acquisition interfaces and optoelectronic components building such force spectroscopy AFM is relatively easy. However, suitable software to control such laboratory made instrument was not released. Developing it in the lab requires significant time and effort. Our Nanopuller software described in this paper is intended to eliminate this obstacle. With only minimum adjustments this program can be used to control and acquire data with any suitable National Instruments universal digital/analog interface and piezoelectric actuator analog controller, giving significant freedom and flexibility in designing force spectroscopy experiment. Since the full code, written in a graphical LabVIEW environment is available, our Nanopuller can be easily customized. In this paper we describe the program and test its performance in controlling different setups. Successful and accurate force curve acquisition for standard samples (single molecules of I27O reference titin polyprotein and DNA as well as red blood cells) is shown. - Highlights: • We created open data acquisition software for performing Atomic Force Microscopy force measurements with custom laboratory made setups. • The software allows large flexibility in atomic force microscope design with minimum adjustment necessary. • The software is written in LabVIEW, allowing easy customization. • We successfully tested the program on two different hardware configurations by stretching single macromolecules and indenting cells.

  20. Taking Nanomedicine Teaching into Practice with Atomic Force Microscopy and Force Spectroscopy

    Carvalho, Filomena A.; Freitas, Teresa; Santos, Nuno C.

    2015-01-01

    Atomic force microscopy (AFM) is a useful and powerful tool to study molecular interactions applied to nanomedicine. The aim of the present study was to implement a hands-on atomic AFM course for graduated biosciences and medical students. The course comprises two distinct practical sessions, where students get in touch with the use of an atomic…

  1. Gas detection by correlation spectroscopy employing a multimode diode laser.

    Lou, Xiutao; Somesfalean, Gabriel; Zhang, Zhiguo

    2008-05-01

    A gas sensor based on the gas-correlation technique has been developed using a multimode diode laser (MDL) in a dual-beam detection scheme. Measurement of CO(2) mixed with CO as an interfering gas is successfully demonstrated using a 1570 nm tunable MDL. Despite overlapping absorption spectra and occasional mode hops, the interfering signals can be effectively excluded by a statistical procedure including correlation analysis and outlier identification. The gas concentration is retrieved from several pair-correlated signals by a linear-regression scheme, yielding a reliable and accurate measurement. This demonstrates the utility of the unsophisticated MDLs as novel light sources for gas detection applications.

  2. Correlation Spectroscopy of Surfaces, Thin Films, and Nanostructures

    Berakdar, Jamal

    2004-01-01

    Here, leading scientists present an overview of the most modern experimental and theoretical methods for studying electronic correlations on surfaces, in thin films and in nanostructures. In particular, they describe in detail coincidence techniques for studying many-particle correlations while. critically examining the informational content of such processes from a theoretical point viewpoint. Furthermore, the book considers the current state of incorporating many-body effects into theoretical approaches. Covered topics:. -Auger-electron photoelectron coincidence experiments and theories. -Co

  3. Unraveling protein-protein interactions in clathrin assemblies via atomic force spectroscopy.

    Jin, Albert J; Lafer, Eileen M; Peng, Jennifer Q; Smith, Paul D; Nossal, Ralph

    2013-03-01

    Atomic force microscopy (AFM), single molecule force spectroscopy (SMFS), and single particle force spectroscopy (SPFS) are used to characterize intermolecular interactions and domain structures of clathrin triskelia and clathrin-coated vesicles (CCVs). The latter are involved in receptor-mediated endocytosis (RME) and other trafficking pathways. Here, we subject individual triskelia, bovine-brain CCVs, and reconstituted clathrin-AP180 coats to AFM-SMFS and AFM-SPFS pulling experiments and apply novel analytics to extract force-extension relations from very large data sets. The spectroscopic fingerprints of these samples differ markedly, providing important new information about the mechanism of CCV uncoating. For individual triskelia, SMFS reveals a series of events associated with heavy chain alpha-helix hairpin unfolding, as well as cooperative unraveling of several hairpin domains. SPFS of clathrin assemblies exposes weaker clathrin-clathrin interactions that are indicative of inter-leg association essential for RME and intracellular trafficking. Clathrin-AP180 coats are energetically easier to unravel than the coats of CCVs, with a non-trivial dependence on force-loading rate. Published by Elsevier Inc.

  4. AFM-based force spectroscopy on polystyrene brushes: effect of brush thickness on protein adsorption.

    Hentschel, Carsten; Wagner, Hendrik; Smiatek, Jens; Heuer, Andreas; Fuchs, Harald; Zhang, Xi; Studer, Armido; Chi, Lifeng

    2013-02-12

    Herein we present a study on nonspecific binding of proteins at highly dense packed hydrophobic polystyrene brushes. In this context, an atomic force microscopy tip was functionalized with concanavalin A to perform single-molecule force spectroscopy measurements on polystyrene brushes with thicknesses of 10 and 60 nm, respectively. Polystyrene brushes with thickness of 10 nm show an almost two times stronger protein adsorption than brushes with a thickness of 60 nm: 72 pN for the thinner and 38 pN for the thicker layer, which is in qualitative agreement with protein adsorption studies conducted macroscopically by fluorescence microscopy.

  5. Communication: atomic force detection of single-molecule nonlinear optical vibrational spectroscopy.

    Saurabh, Prasoon; Mukamel, Shaul

    2014-04-28

    Atomic Force Microscopy (AFM) allows for a highly sensitive detection of spectroscopic signals. This has been first demonstrated for NMR of a single molecule and recently extended to stimulated Raman in the optical regime. We theoretically investigate the use of optical forces to detect time and frequency domain nonlinear optical signals. We show that, with proper phase matching, the AFM-detected signals closely resemble coherent heterodyne-detected signals. Applications are made to AFM-detected and heterodyne-detected vibrational resonances in Coherent Anti-Stokes Raman Spectroscopy (χ((3))) and sum or difference frequency generation (χ((2))).

  6. Principal Component Analysis Based Two-Dimensional (PCA-2D) Correlation Spectroscopy: PCA Denoising for 2D Correlation Spectroscopy

    Jung, Young Mee

    2003-01-01

    Principal component analysis based two-dimensional (PCA-2D) correlation analysis is applied to FTIR spectra of polystyrene/methyl ethyl ketone/toluene solution mixture during the solvent evaporation. Substantial amount of artificial noise were added to the experimental data to demonstrate the practical noise-suppressing benefit of PCA-2D technique. 2D correlation analysis of the reconstructed data matrix from PCA loading vectors and scores successfully extracted only the most important features of synchronicity and asynchronicity without interference from noise or insignificant minor components. 2D correlation spectra constructed with only one principal component yield strictly synchronous response with no discernible a asynchronous features, while those involving at least two or more principal components generated meaningful asynchronous 2D correlation spectra. Deliberate manipulation of the rank of the reconstructed data matrix, by choosing the appropriate number and type of PCs, yields potentially more refined 2D correlation spectra

  7. Quantifying bacterial adhesion on antifouling polymer brushes via single-cell force spectroscopy

    Rodriguez-Emmenegger, Cesar; Janel, S.; de los Santos Pereira, Andres; Bruns, M.; Lafont, F.

    2015-01-01

    Roč. 6, č. 31 (2015), s. 5740-5751 ISSN 1759-9954 R&D Projects: GA ČR(CZ) GJ15-09368Y; GA MŠk(CZ) ED1.1.00/02.0109 Grant - others:OPPK(XE) CZ.2.16/3.1.00/21545 Program:OPPK Institutional support: RVO:61389013 Keywords : antifouling polymer brushes * single-cell force spectroscopy * bacterial adhesion Subject RIV: BO - Biophysics Impact factor: 5.687, year: 2015

  8. 1H-15N correlation spectroscopy of nanocrystalline proteins

    Morcombe, Corey R.; Paulson, Eric K.; Gaponenko, Vadim; Byrd, R. Andrew; Zilm, Kurt W.

    2005-01-01

    The limits of resolution that can be obtained in 1 H- 15 N 2D NMR spectroscopy of isotopically enriched nanocrystalline proteins are explored. Combinations of frequency switched Lee-Goldburg (FSLG) decoupling, fast magic angle sample spinning (MAS), and isotopic dilution via deuteration are investigated as methods for narrowing the amide 1 H resonances. Heteronuclear decoupling of 15 N from the 1 H resonances is also studied. Using human ubiquitin as a model system, the best resolution is most easily obtained with uniformly 2 H and 15 N enriched protein where the amides have been exchanged in normal water, MAS at ∼20 kHz, and WALTZ-16 decoupling of the 15 N nuclei. The combination of these techniques results in average 1 H lines of only ∼0.26 ppm full width at half maximum. Techniques for optimizing instrument stability and 15 N decoupling are described for achieving the best possible performance in these experiments

  9. Magnetic resonance spectroscopy of normal appearing white matter in early relapsing-remitting multiple sclerosis: correlations between disability and spectroscopy

    Foronda Jesus

    2004-06-01

    Full Text Available Abstract Background What currently appears to be irreversible axonal loss in normal appearing white matter, measured by proton magnetic resonance spectroscopy is of great interest in the study of Multiple Sclerosis. Our aim is to determine the axonal damage in normal appearing white matter measured by magnetic resonance spectroscopy and to correlate this with the functional disability measured by Multiple Sclerosis Functional Composite scale, Neurological Rating Scale, Ambulation Index scale, and Expanded Disability Scale Score. Methods Thirty one patients (9 male and 22 female with relapsing remitting Multiple Sclerosis and a Kurtzke Expanded Disability Scale Score of 0–5.5 were recruited from four hospitals in Andalusia, Spain and included in the study. Magnetic resonance spectroscopy scans and neurological disability assessments were performed the same day. Results A statistically significant correlation was found (r = -0.38 p Conclusions There is correlation between disability (measured by Expanded Disability Scale Score and the NAA/Cr ratio in normal appearing white matter. The lack of correlation between the NAA/Cr ratio and the Multiple Sclerosis Functional Composite score indicates that the Multiple Sclerosis Functional Composite is not able to measure irreversible disability and would be more useful as a marker in stages where axonal damage is not a predominant factor.

  10. Study on Senna alata and its different extracts by Fourier transform infrared spectroscopy and two-dimensional correlation infrared spectroscopy

    Adiana, M. A.; Mazura, M. P.

    2011-04-01

    Senna alata L. commonly known as candle bush belongs to the family of Fabaceae and the plant has been reported to possess anti-inflammatory, analgesic, laxative and antiplatelet-aggregating activity. In order to develop a rapid and effective analysis method for studying integrally the main constituents in the medicinal materials and their extracts, discriminating the extracts from different extraction process, comparing the categories of chemical constituents in the different extracts and monitoring the qualities of medicinal materials, we applied Fourier transform infrared spectroscopy (FT-IR) associated with second derivative infrared spectroscopy and two-dimensional infrared correlation spectroscopy (2D-IR) to study the main constituents of S. alata and its different extracts (extracted by hexane, dichloromethane, ethyl acetate and methanol in turn). The findings indicated that FT-IR and 2D-IR can provide many holistic variation rules of chemical constituents. Use of the macroscopical fingerprint characters of FT-IR and 2D-IR spectrum can identify the main chemical constituents in medicinal materials and their extracts, but also compare the components differences among similar samples. In a conclusion, FT-IR spectroscopy combined with 2D correlation analysis provides a powerful method for the quality control of traditional medicines.

  11. Peculiarities of plasma homeostasis in the patients with rectal cancer according to laser correlation spectroscopy findings

    Byilenko, O.A.; Bazhora, Yu.Yi.; Sokolov, V.M.; Andronov, D.Yu.

    1997-01-01

    Laser correlation spectroscopy was used to investigate plasma homeostasis in 82 patients with rectal cancer. The spectra of the blood plasma from 21 donors of the transfusion station were used as the control. The blood plasma homeostasis changes reheated with laser correlation spectrometry in the patients with rectal cancer allow to use them for diagnosis of this pathology

  12. Superheavy-element spectroscopy: Correlations along element 115 decay chains

    Rudolph, D.; Forsberg, U.; Sarmiento, L. G.; Golubev, P.; Fahlander, C.

    2016-05-01

    Following a brief summary of the region of the heaviest atomic nuclei yet created in the laboratory, data on more than hundred α-decay chains associated with the production of element 115 are combined to investigate time and energy correlations along the observed decay chains. Several of these are analysed using a new method for statistical assessments of lifetimes in sets of decay chains.

  13. Force spectroscopy studies on protein-ligand interactions: a single protein mechanics perspective.

    Hu, Xiaotang; Li, Hongbin

    2014-10-01

    Protein-ligand interactions are ubiquitous and play important roles in almost every biological process. The direct elucidation of the thermodynamic, structural and functional consequences of protein-ligand interactions is thus of critical importance to decipher the mechanism underlying these biological processes. A toolbox containing a variety of powerful techniques has been developed to quantitatively study protein-ligand interactions in vitro as well as in living systems. The development of atomic force microscopy-based single molecule force spectroscopy techniques has expanded this toolbox and made it possible to directly probe the mechanical consequence of ligand binding on proteins. Many recent experiments have revealed how ligand binding affects the mechanical stability and mechanical unfolding dynamics of proteins, and provided mechanistic understanding on these effects. The enhancement effect of mechanical stability by ligand binding has been used to help tune the mechanical stability of proteins in a rational manner and develop novel functional binding assays for protein-ligand interactions. Single molecule force spectroscopy studies have started to shed new lights on the structural and functional consequence of ligand binding on proteins that bear force under their biological settings. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  14. A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

    Gong, S.; Labanca, I.; Rech, I.; Ghioni, M. [Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy)

    2014-10-15

    Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds.

  15. A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

    Gong, S.; Labanca, I.; Rech, I.; Ghioni, M.

    2014-01-01

    Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds

  16. AFM-based force spectroscopy measurements of mature amyloid fibrils of the peptide glucagon

    Dong, M. D.; Hovgaard, M. B.; Mamdouh, W.

    2008-01-01

    We report on the mechanical characterization of individual mature amyloid fibrils by atomic force microscopy (AFM) and AFM-based single-molecule force spectroscopy (SMFS). These self-assembling materials, formed from the 29-residue amphiphatic peptide hormone glucagon, were found to display...... a reversible elastic behaviour. Based on AFM morphology and SMFS studies, we suggest that the observed elasticity is due to a force-induced conformational transition which is reversible due to the beta-helical conformation of protofibrils, allowing a high degree of extension. The elastic properties...... of such mature fibrils contribute to their high stability, suggesting that the internal hydrophobic interactions of amyloid fibrils are likely to be of fundamental importance in the assembly of amyloid fibrils and therefore for the understanding of the progression of their associated pathogenic disorders...

  17. Force spectroscopy of collagen fibers to investigate their mechanical properties and structural organization.

    Gutsmann, Thomas; Fantner, Georg E; Kindt, Johannes H; Venturoni, Manuela; Danielsen, Signe; Hansma, Paul K

    2004-05-01

    Tendons are composed of collagen and other molecules in a highly organized hierarchical assembly, leading to extraordinary mechanical properties. To probe the cross-links on the lower level of organization, we used a cantilever to pull substructures out of the assembly. Advanced force probe technology, using small cantilevers (length exponential increase in force and two different periodic rupture events, one with strong bonds (jumps in force of several hundred pN) with a periodicity of 78 nm and one with weak bonds (jumps in force of <7 pN) with a periodicity of 22 nm. We demonstrate a good correlation between the measured mechanical behavior of collagen fibers and their appearance in the micrographs taken with the atomic force microscope.

  18. Influence of effective three-body force on the spectroscopy of 19O

    Haung, W.; Song, H.; Wang, Z.; Kuo, T.T.S.

    1983-01-01

    The purpose of the present paper is to investigate the influence of effective three-body force on the spectroscopy of 19 O. The model space was chosen as the configuration space which consists of the j-j coupling states of three valence neutrons in the s-d shell. The effective interaction including two- and three-body forces was then derived in the framework of the folded diagram method (FDM). Besides two traditional three-body terms, there is another kind of three-body force, the folded one constructed with two two-body diagrams, in FDM. The G-matrix elements of soft core Reid force were used in the numerical calculations. In the case of lacking the G-matrix elements, we adopted the matrix elements of M-3Y force as the equivalents. The results show that the influence of the effective three-body forces on the spectrum of 19 O is not of importance, but the part coming from the folded three-body term is worth noting

  19. Characterisation of cellular adhesion reinforcement by multiple bond force spectroscopy in alveolar epithelial cells.

    Nguyen, Ngoc-Minh; Angely, Christelle; Andre Dias, Sofia; Planus, Emmanuelle; Filoche, Marcel; Pelle, Gabriel; Louis, Bruno; Isabey, Daniel

    2017-07-01

    Integrin-mediated adhesion is a key process by which cells physically connect with their environment, and express sensitivity and adaptation through mechanotransduction. A critical step of cell adhesion is the formation of the first bonds which individually generate weak contacts (∼tens pN) but can sustain thousand times higher forces (∼tens nN) when associated. We propose an experimental validation by multiple bond force spectroscopy (MFS) of a stochastic model predicting adhesion reinforcement permitted by non-cooperative, multiple bonds on which force is homogeneously distributed (called parallel bond configuration). To do so, spherical probes (diameter: 6.6 μm), specifically coated by RGD-peptide to bind integrins, are used to statically indent and homogenously stretch the multiple bonds created for short contact times (2 s) between the bead and the surface of epithelial cells (A549). Using different separation speeds (v = 2, 5, 10 μm/s) and measuring cellular Young's modulus as well as the local stiffness preceding local rupture events, we obtain cell-by-cell the effective loading rates both at the global cell level and at the local level of individual constitutive bonds. Local rupture forces are in the range: f*=60-115 pN , whereas global rupture (detachment) forces reach F*=0.8-1.7 nN . Global and local rupture forces both exhibit linear dependencies with the effective loading rate, the slopes of these two linear relationships providing an estimate of the number of independent integrin bonds constituting the tested multiple bond structure (∼12). The MFS method enables to validate the reinforcement of integrin-mediated adhesion induced by the multiple bond configuration in which force is homogeneously distributed amongst parallel bonds. Local rupture events observed in the course of a spectroscopy manoeuver (MFS) lead to rupture force values considered in the literature as single-integrin bonds. Adhesion reinforcement permitted by the parallel

  20. Reduced density-matrix functional theory: Correlation and spectroscopy.

    Di Sabatino, S; Berger, J A; Reining, L; Romaniello, P

    2015-07-14

    In this work, we explore the performance of approximations to electron correlation in reduced density-matrix functional theory (RDMFT) and of approximations to the observables calculated within this theory. Our analysis focuses on the calculation of total energies, occupation numbers, removal/addition energies, and spectral functions. We use the exactly solvable Hubbard dimer at 1/4 and 1/2 fillings as test systems. This allows us to analyze the underlying physics and to elucidate the origin of the observed trends. For comparison, we also report the results of the GW approximation, where the self-energy functional is approximated, but no further hypothesis is made concerning the approximations of the observables. In particular, we focus on the atomic limit, where the two sites of the dimer are pulled apart and electrons localize on either site with equal probability, unless a small perturbation is present: this is the regime of strong electron correlation. In this limit, using the Hubbard dimer at 1/2 filling with or without a spin-symmetry-broken ground state allows us to explore how degeneracies and spin-symmetry breaking are treated in RDMFT. We find that, within the used approximations, neither in RDMFT nor in GW, the signature of strong correlation is present, when looking at the removal/addition energies and spectral function from the spin-singlet ground state, whereas both give the exact result for the spin-symmetry broken case. Moreover, we show how the spectroscopic properties change from one spin structure to the other.

  1. Knee adduction moment and medial contact force--facts about their correlation during gait.

    Ines Kutzner

    Full Text Available The external knee adduction moment is considered a surrogate measure for the medial tibiofemoral contact force and is commonly used to quantify the load reducing effect of orthopedic interventions. However, only limited and controversial data exist about the correlation between adduction moment and medial force. The objective of this study was to examine whether the adduction moment is indeed a strong predictor for the medial force by determining their correlation during gait. Instrumented knee implants with telemetric data transmission were used to measure tibiofemoral contact forces in nine subjects. Gait analyses were performed simultaneously to the joint load measurements. Skeletal kinematics, as well as the ground reaction forces and inertial parameters, were used as inputs in an inverse dynamics approach to calculate the external knee adduction moment. Linear regression analysis was used to analyze the correlation between adduction moment and medial force for the whole stance phase and separately for the early and late stance phase. Whereas only moderate correlations between adduction moment and medial force were observed throughout the whole stance phase (R(2 = 0.56 and during the late stance phase (R(2 = 0.51, a high correlation was observed at the early stance phase (R(2 = 0.76. Furthermore, the adduction moment was highly correlated to the medial force ratio throughout the whole stance phase (R(2 = 0.75. These results suggest that the adduction moment is a surrogate measure, well-suited to predicting the medial force ratio throughout the whole stance phase or medial force during the early stance phase. However, particularly during the late stance phase, moderate correlations and high inter-individual variations revealed that the predictive value of the adduction moment is limited. Further analyses are necessary to examine whether a combination of other kinematic, kinetic or neuromuscular factors may lead to a more

  2. Sensing of silver nanoparticles on/in endothelial cells using atomic force spectroscopy.

    Kolodziejczyk, Agnieszka; Jakubowska, Aleksandra; Kucinska, Magdalena; Wasiak, Tomasz; Komorowski, Piotr; Makowski, Krzysztof; Walkowiak, Bogdan

    2018-05-10

    Endothelial cells, due to their location, are interesting objects for atomic force spectroscopy study. They constitute a barrier between blood and vessel tissues located deeper, and therefore they are the first line of contact with various substances present in blood, eg, drugs or nanoparticles. This work intends to verify whether the mechanical response of immortalized human umbilical vein endothelial cells (EA.hy926), when exposed to silver nanoparticles, as measured using force spectroscopy, could be effectively used as a bio-indicator of the physiological state of the cells. Silver nanoparticles were characterized with transmission electron microscopy and dynamic light scattering techniques. Tetrazolium salt reduction test was used to determine cell viability after treatment with silver nanoparticles. An elasticity of native cells was examined in the Hanks' buffer whereas fixed cells were softly fixed with formaldehyde. Additional aspect of the work is the comparative force spectroscopy utilizing AFM probes of ball-shape and conical geometries, in order to understand what changes in cell elasticity, caused by SNPs, were detectable with each probe. As a supplement to elasticity studies, cell morphology observation by atomic force microscopy and detection of silver nanoparticles inside cells using transmission electron microscopy were also performed. Cells exposed to silver nanoparticles at the highest selected concentrations (3.6 μg/mL, 16 μg/mL) are less elastic. It may be associated with the reorganization of the cellular cytoskeleton and the "strengthening" of the cell cortex caused by presence of silver nanoparticles. This observation does not depend on cell fixation. Agglomerates of silver nanoparticles were observed on the cell membrane as well as inside the cells. Copyright © 2018 John Wiley & Sons, Ltd.

  3. Two-dimensional optical correlation spectroscopy applied to liquid/glass dynamics

    Lazonder, Kees; Pshenichnikov, Maxim S.; Wiersma, Douwe A.; Corkum, Paul; Jonas, David M.; Miller, R.J. Dwayne.; Weiner, Andrew M.

    2007-01-01

    Correlation spectroscopy was used to study the effects of temperature and phase changes on liquid and glass solvent dynamics. By assessing the eccentricity of the elliptic shape of a 2D optical correlation spectrum the value of the underlying frequency-frequency correlation function can be retrieved through a very simple relationship. This method yielded both intuitive clues and a quantitative measure of the dynamics of the system.

  4. Dielectrophoretic positioning of single nanoparticles on atomic force microscope tips for tip-enhanced Raman spectroscopy.

    Leiterer, Christian; Deckert-Gaudig, Tanja; Singh, Prabha; Wirth, Janina; Deckert, Volker; Fritzsche, Wolfgang

    2015-05-01

    Tip-enhanced Raman spectroscopy, a combination of Raman spectroscopy and scanning probe microscopy, is a powerful technique to detect the vibrational fingerprint of molecules at the nanometer scale. A metal nanoparticle at the apex of an atomic force microscope tip leads to a large enhancement of the electromagnetic field when illuminated with an appropriate wavelength, resulting in an increased Raman signal. A controlled positioning of individual nanoparticles at the tip would improve the reproducibility of the probes and is quite demanding due to usually serial and labor-intensive approaches. In contrast to commonly used submicron manipulation techniques, dielectrophoresis allows a parallel and scalable production, and provides a novel approach toward reproducible and at the same time affordable tip-enhanced Raman spectroscopy tips. We demonstrate the successful positioning of an individual plasmonic nanoparticle on a commercial atomic force microscope tip by dielectrophoresis followed by experimental proof of the Raman signal enhancing capabilities of such tips. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Optimizing 1-μs-Resolution Single-Molecule Force Spectroscopy on a Commercial Atomic Force Microscope.

    Edwards, Devin T; Faulk, Jaevyn K; Sanders, Aric W; Bull, Matthew S; Walder, Robert; LeBlanc, Marc-Andre; Sousa, Marcelo C; Perkins, Thomas T

    2015-10-14

    Atomic force microscopy (AFM)-based single-molecule force spectroscopy (SMFS) is widely used to mechanically measure the folding and unfolding of proteins. However, the temporal resolution of a standard commercial cantilever is 50-1000 μs, masking rapid transitions and short-lived intermediates. Recently, SMFS with 0.7-μs temporal resolution was achieved using an ultrashort (L = 9 μm) cantilever on a custom-built, high-speed AFM. By micromachining such cantilevers with a focused ion beam, we optimized them for SMFS rather than tapping-mode imaging. To enhance usability and throughput, we detected the modified cantilevers on a commercial AFM retrofitted with a detection laser system featuring a 3-μm circular spot size. Moreover, individual cantilevers were reused over multiple days. The improved capabilities of the modified cantilevers for SMFS were showcased by unfolding a polyprotein, a popular biophysical assay. Specifically, these cantilevers maintained a 1-μs response time while eliminating cantilever ringing (Q ≅ 0.5). We therefore expect such cantilevers, along with the instrumentational improvements to detect them on a commercial AFM, to accelerate high-precision AFM-based SMFS studies.

  6. Investigating the binding behaviour of two avidin-based testosterone binders using molecular recognition force spectroscopy.

    Rangl, Martina; Leitner, Michael; Riihimäki, Tiina; Lehtonen, Soili; Hytönen, Vesa P; Gruber, Hermann J; Kulomaa, Markku; Hinterdorfer, Peter; Ebner, Andreas

    2014-02-01

    Molecular recognition force spectroscopy, a biosensing atomic force microscopy technique allows to characterise the dissociation of ligand-receptor complexes at the molecular level. Here, we used molecular recognition force spectroscopy to study the binding capability of recently developed testosterone binders. The two avidin-based proteins called sbAvd-1 and sbAvd-2 are expected to bind both testosterone and biotin but differ in their binding behaviour towards these ligands. To explore the ligand binding and dissociation energy landscape of these proteins, we tethered biotin or testosterone to the atomic force microscopy probe while the testosterone-binding protein was immobilized on the surface. Repeated formation and rupture of the ligand-receptor complex at different pulling velocities allowed determination of the loading rate dependence of the complex-rupturing force. In this way, we obtained the molecular dissociation rate (k(off)) and energy landscape distances (x(β)) of the four possible complexes: sbAvd-1-biotin, sbAvd-1-testosterone, sbAvd-2-biotin and sbAvd-2-testosterone. It was found that the kinetic off-rates for both proteins and both ligands are similar. In contrast, the x(β) values, as well as the probability of complex formations, varied considerably. In addition, competitive binding experiments with biotin and testosterone in solution differ significantly for the two testosterone-binding proteins, implying a decreased cross-reactivity of sbAvd-2. Unravelling the binding behaviour of the investigated testosterone-binding proteins is expected to improve their usability for possible sensing applications. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Single-cell force spectroscopy as a technique to quantify human red blood cell adhesion to subendothelial laminin.

    Maciaszek, Jamie L; Partola, Kostyantyn; Zhang, Jing; Andemariam, Biree; Lykotrafitis, George

    2014-12-18

    Single-cell force spectroscopy (SCFS), an atomic force microscopy (AFM)-based assay, enables quantitative study of cell adhesion while maintaining the native state of surface receptors in physiological conditions. Human healthy and pathological red blood cells (RBCs) express a large number of surface proteins which mediate cell-cell interactions, or cell adhesion to the extracellular matrix. In particular, RBCs adhere with high affinity to subendothelial matrix laminin via the basal cell adhesion molecule and Lutheran protein (BCAM/Lu). Here, we established SCFS as an in vitro technique to study human RBC adhesion at baseline and following biochemical treatment. Using blood obtained from healthy human subjects, we recorded adhesion forces from single RBCs attached to AFM cantilevers as the cell was pulled-off of substrates coated with laminin protein. We found that an increase in the overall cell adhesion measured via SCFS is correlated with an increase in the resultant total force measured on 1 µm(2) areas of the RBC membrane. Further, we showed that SCFS can detect significant changes in the adhesive response of RBCs to modulation of the cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) pathway. Lastly, we identified variability in the RBC adhesion force to laminin amongst the human subjects, suggesting that RBCs maintain diverse levels of active BCAM/Lu adhesion receptors. By using single-cell measurements, we established a powerful new method for the quantitative measurement of single RBC adhesion with specific receptor-mediated binding. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Fluorescence correlation spectroscopy: Ultrasensitive detection in clear and turbid media

    Tahari, Abdel Kader

    In this work, I describe the development of a simple, inexpensive, and powerful alternative technique to detect and analyze, without enrichment, extremely low concentrations of cells, bacteria, viruses, and protein aggregates in turbid fluids for clinical and biotechnological applications. The anticipated applications of this technique are many. They range from the determination of the somatic cell count in milk for the dairy industry, to the enumeration and characterization of microorganisms in environmental microbiology and the food industry, and to the fast and ultrasensitive detection of protein aggregates for the diagnosis of Alzheimer's and other neurodegenerative diseases in clinical medicine. A prototype instrument has been built and allowed the detection and quantification of particles down to a few per milliliter in short scanning times. It consists of a small microscope that has a horizontal geometry and a mechanical instrument that holds a cylindrical cuvette (1 cm in diameter) with two motors that provide a rotational and a slower vertical inversion motions. The illumination focus is centered about 200 mum from the wall of the cuvette inside the sample. The total volume that is explored is large (˜1ml/min for bright particles). The data is analyzed with a correlation filter program based on particle passage pattern recognition. I will also describe further work on improving the sensitivity of the technique, expanding it for multiple-species discrimination and enumeration, and testing the prototype device in actual clinical and biotechnological applications. The main clinical application of this project seeks to establish conditions and use this new technique to quantify and size-analyze oligomeric complexes of the Alzheimer's disease beta-peptide in cerebrospinal fluid and other body fluids as a molecular biomarker for persons at risk of Alzheimer's disease dementia. The technology could potentially be extended to the diagnosis and therapeutic

  9. Elastic Properties of Nucleic Acids by Single-Molecule Force Spectroscopy.

    Camunas-Soler, Joan; Ribezzi-Crivellari, Marco; Ritort, Felix

    2016-07-05

    We review the current knowledge on the use of single-molecule force spectroscopy techniques to extrapolate the elastic properties of nucleic acids. We emphasize the lesser-known elastic properties of single-stranded DNA. We discuss the importance of accurately determining the elastic response in pulling experiments, and we review the simplest models used to rationalize the experimental data as well as the experimental approaches used to pull single-stranded DNA. Applications used to investigate DNA conformational transitions and secondary structure formation are also highlighted. Finally, we provide an overview of the effects of salt and temperature and briefly discuss the effects of contour length and sequence dependence.

  10. Correlates of Forced Sex Among Populations of Men Who Have Sex with Men in Thailand

    Guadamuz, Thomas E.; Wimonsate, Wipas; Varangrat, Anchalee; Phanuphak, Praphan; Jommaroeng, Rapeepun; Mock, Philip A.; Tappero, Jordan W.; van Griensven, Frits

    2009-01-01

    Although forced sex is a correlate of HIV infection, its prevalence and associated risks are not well described among men who have sex with men (MSM) in developing-country settings. Between March and October 2005, we assessed the prevalence of forced sex and correlates among populations of MSM (this includes general MSM, male sex workers, and male-to-female transgender persons) in Thailand using a community-based sample. Participants were enrolled from venues around Bangkok, Chiangmai, and Ph...

  11. Atomic force microscopy and spectroscopy to probe single membrane proteins in lipid bilayers.

    Sapra, K Tanuj

    2013-01-01

    The atomic force microscope (AFM) has opened vast avenues hitherto inaccessible to the biological scientist. The high temporal (millisecond) and spatial (nanometer) resolutions of the AFM are suited for studying many biological processes in their native conditions. The AFM cantilever stylus is aptly termed as a "lab on a tip" owing to its versatility as an imaging tool as well as a handle to manipulate single bonds and proteins. Recent examples assert that the AFM can be used to study the mechanical properties and monitor processes of single proteins and single cells, thus affording insight into important mechanistic details. This chapter specifically focuses on practical and analytical protocols of single-molecule AFM methodologies related to high-resolution imaging and single-molecule force spectroscopy of membrane proteins. Both these techniques are operator oriented, and require specialized working knowledge of the instrument, theoretical, and practical skills.

  12. Elastin-like Polypeptide Linkers for Single-Molecule Force Spectroscopy.

    Ott, Wolfgang; Jobst, Markus A; Bauer, Magnus S; Durner, Ellis; Milles, Lukas F; Nash, Michael A; Gaub, Hermann E

    2017-06-27

    Single-molecule force spectroscopy (SMFS) is by now well established as a standard technique in biophysics and mechanobiology. In recent years, the technique has benefitted greatly from new approaches to bioconjugation of proteins to surfaces. Indeed, optimized immobilization strategies for biomolecules and refined purification schemes are being steadily adapted and improved, which in turn has enhanced data quality. In many previously reported SMFS studies, poly(ethylene glycol) (PEG) was used to anchor molecules of interest to surfaces and/or cantilever tips. The limitation, however, is that PEG exhibits a well-known trans-trans-gauche to all-trans transition, which results in marked deviation from standard polymer elasticity models such as the worm-like chain, particularly at elevated forces. As a result, the assignment of unfolding events to protein domains based on their corresponding amino acid chain lengths is significantly obscured. Here, we provide a solution to this problem by implementing unstructured elastin-like polypeptides as linkers to replace PEG. We investigate the suitability of tailored elastin-like polypeptides linkers and perform direct comparisons to PEG, focusing on attributes that are critical for single-molecule force experiments such as linker length, monodispersity, and bioorthogonal conjugation tags. Our results demonstrate that by avoiding the ambiguous elastic response of mixed PEG/peptide systems and instead building the molecular mechanical systems with only a single bond type with uniform elastic properties, we improve data quality and facilitate data analysis and interpretation in force spectroscopy experiments. The use of all-peptide linkers allows alternative approaches for precisely defining elastic properties of proteins linked to surfaces.

  13. High-throughput single-molecule force spectroscopy for membrane proteins

    Bosshart, Patrick D.; Casagrande, Fabio; Frederix, Patrick L. T. M.; Ratera, Merce; Bippes, Christian A.; Müller, Daniel J.; Palacin, Manuel; Engel, Andreas; Fotiadis, Dimitrios

    2008-09-01

    Atomic force microscopy-based single-molecule force spectroscopy (SMFS) is a powerful tool for studying the mechanical properties, intermolecular and intramolecular interactions, unfolding pathways, and energy landscapes of membrane proteins. One limiting factor for the large-scale applicability of SMFS on membrane proteins is its low efficiency in data acquisition. We have developed a semi-automated high-throughput SMFS (HT-SMFS) procedure for efficient data acquisition. In addition, we present a coarse filter to efficiently extract protein unfolding events from large data sets. The HT-SMFS procedure and the coarse filter were validated using the proton pump bacteriorhodopsin (BR) from Halobacterium salinarum and the L-arginine/agmatine antiporter AdiC from the bacterium Escherichia coli. To screen for molecular interactions between AdiC and its substrates, we recorded data sets in the absence and in the presence of L-arginine, D-arginine, and agmatine. Altogether ~400 000 force-distance curves were recorded. Application of coarse filtering to this wealth of data yielded six data sets with ~200 (AdiC) and ~400 (BR) force-distance spectra in each. Importantly, the raw data for most of these data sets were acquired in one to two days, opening new perspectives for HT-SMFS applications.

  14. High-throughput single-molecule force spectroscopy for membrane proteins

    Bosshart, Patrick D; Casagrande, Fabio; Frederix, Patrick L T M; Engel, Andreas; Fotiadis, Dimitrios; Ratera, Merce; Palacin, Manuel; Bippes, Christian A; Mueller, Daniel J

    2008-01-01

    Atomic force microscopy-based single-molecule force spectroscopy (SMFS) is a powerful tool for studying the mechanical properties, intermolecular and intramolecular interactions, unfolding pathways, and energy landscapes of membrane proteins. One limiting factor for the large-scale applicability of SMFS on membrane proteins is its low efficiency in data acquisition. We have developed a semi-automated high-throughput SMFS (HT-SMFS) procedure for efficient data acquisition. In addition, we present a coarse filter to efficiently extract protein unfolding events from large data sets. The HT-SMFS procedure and the coarse filter were validated using the proton pump bacteriorhodopsin (BR) from Halobacterium salinarum and the L-arginine/agmatine antiporter AdiC from the bacterium Escherichia coli. To screen for molecular interactions between AdiC and its substrates, we recorded data sets in the absence and in the presence of L-arginine, D-arginine, and agmatine. Altogether ∼400 000 force-distance curves were recorded. Application of coarse filtering to this wealth of data yielded six data sets with ∼200 (AdiC) and ∼400 (BR) force-distance spectra in each. Importantly, the raw data for most of these data sets were acquired in one to two days, opening new perspectives for HT-SMFS applications

  15. High-throughput single-molecule force spectroscopy for membrane proteins

    Bosshart, Patrick D; Casagrande, Fabio; Frederix, Patrick L T M; Engel, Andreas; Fotiadis, Dimitrios [M E Mueller Institute for Structural Biology, Biozentrum of the University of Basel, CH-4056 Basel (Switzerland); Ratera, Merce; Palacin, Manuel [Institute for Research in Biomedicine, Barcelona Science Park, Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona and Centro de Investigacion Biomedica en Red de Enfermedades Raras, E-08028 Barcelona (Spain); Bippes, Christian A; Mueller, Daniel J [BioTechnology Center, Technical University, Tatzberg 47, D-01307 Dresden (Germany)], E-mail: andreas.engel@unibas.ch, E-mail: dimitrios.fotiadis@mci.unibe.ch

    2008-09-24

    Atomic force microscopy-based single-molecule force spectroscopy (SMFS) is a powerful tool for studying the mechanical properties, intermolecular and intramolecular interactions, unfolding pathways, and energy landscapes of membrane proteins. One limiting factor for the large-scale applicability of SMFS on membrane proteins is its low efficiency in data acquisition. We have developed a semi-automated high-throughput SMFS (HT-SMFS) procedure for efficient data acquisition. In addition, we present a coarse filter to efficiently extract protein unfolding events from large data sets. The HT-SMFS procedure and the coarse filter were validated using the proton pump bacteriorhodopsin (BR) from Halobacterium salinarum and the L-arginine/agmatine antiporter AdiC from the bacterium Escherichia coli. To screen for molecular interactions between AdiC and its substrates, we recorded data sets in the absence and in the presence of L-arginine, D-arginine, and agmatine. Altogether {approx}400 000 force-distance curves were recorded. Application of coarse filtering to this wealth of data yielded six data sets with {approx}200 (AdiC) and {approx}400 (BR) force-distance spectra in each. Importantly, the raw data for most of these data sets were acquired in one to two days, opening new perspectives for HT-SMFS applications.

  16. Local detection of X-ray spectroscopies with an in-situ Atomic Force Microscope

    Rodrigues, M S; Dhez, O; Denmat, S Le; Felici, R; Comin, F; Chevrier, J

    2008-01-01

    The in situ combination of Scanning Probe Microscopies with X-ray microbeams adds a variety of new possibilities to the panoply of synchrotron radiation techniques. This paper describes an optics-free Atomic Force Microscope that can be directly installed on most of the synchrotron radiation end-stations for combined X-ray and atomic force microscopy experiments. The instrument can be used for atomic force imaging of the investigated sample or to locally measure the X-ray absorption or diffraction, or it can also be used to mechanically interact with the sample while simultaneously taking spectroscopy or diffraction measurements. The local character of these measurements is intrinsically linked with the use of the Atomic Force Microscope tip. It is the sharp tip that gives the opportunity to measure the photons flux impinging on it, or to locally measure the absorption coefficient or the shape of the diffraction pattern. At the end an estimation of the limits of the various techniques presented is also discussed.

  17. Probing static disorder in Arrhenius kinetics by single-molecule force spectroscopy.

    Kuo, Tzu-Ling; Garcia-Manyes, Sergi; Li, Jingyuan; Barel, Itay; Lu, Hui; Berne, Bruce J; Urbakh, Michael; Klafter, Joseph; Fernández, Julio M

    2010-06-22

    The widely used Arrhenius equation describes the kinetics of simple two-state reactions, with the implicit assumption of a single transition state with a well-defined activation energy barrier DeltaE, as the rate-limiting step. However, it has become increasingly clear that the saddle point of the free-energy surface in most reactions is populated by ensembles of conformations, leading to nonexponential kinetics. Here we present a theory that generalizes the Arrhenius equation to include static disorder of conformational degrees of freedom as a function of an external perturbation to fully account for a diverse set of transition states. The effect of a perturbation on static disorder is best examined at the single-molecule level. Here we use force-clamp spectroscopy to study the nonexponential kinetics of single ubiquitin proteins unfolding under force. We find that the measured variance in DeltaE shows both force-dependent and independent components, where the force-dependent component scales with F(2), in excellent agreement with our theory. Our study illustrates a novel adaptation of the classical Arrhenius equation that accounts for the microscopic origins of nonexponential kinetics, which are essential in understanding the rapidly growing body of single-molecule data.

  18. Antigen-antibody biorecognition events as discriminated by noise analysis of force spectroscopy curves.

    Bizzarri, Anna Rita; Cannistraro, Salvatore

    2014-08-22

    Atomic force spectroscopy is able to extract kinetic and thermodynamic parameters of biomolecular complexes provided that the registered unbinding force curves could be reliably attributed to the rupture of the specific complex interactions. To this aim, a commonly used strategy is based on the analysis of the stretching features of polymeric linkers which are suitably introduced in the biomolecule-substrate immobilization procedure. Alternatively, we present a method to select force curves corresponding to specific biorecognition events, which relies on a careful analysis of the force fluctuations of the biomolecule-functionalized cantilever tip during its approach to the partner molecules immobilized on a substrate. In the low frequency region, a characteristic 1/f (α) noise with α equal to one (flickering noise) is found to replace white noise in the cantilever fluctuation power spectrum when, and only when, a specific biorecognition process between the partners occurs. The method, which has been validated on a well-characterized antigen-antibody complex, represents a fast, yet reliable alternative to the use of linkers which may involve additional surface chemistry and reproducibility concerns.

  19. Commercial counterboard for 10 ns software correlator for photon and fluorescence correlation spectroscopy

    Molteni, Matteo; Ferri, Fabio

    2016-11-01

    A 10 ns time resolution, multi-tau software correlator, capable of computing simultaneous autocorrelation (A-A, B-B) and cross (A-B) correlation functions at count rates up to ˜10 MHz, with no data loss, has been developed in LabVIEW and C++ by using the National Instrument timer/counterboard (NI PCIe-6612) and a fast Personal Computer (PC) (Intel Core i7-4790 Processor 3.60 GHz ). The correlator works by using two algorithms: for large lag times (τ ≳ 1 μs), a classical time-mode scheme, based on the measure of the number of pulses per time interval, is used; differently, for τ ≲ 1 μs a photon-mode (PM) scheme is adopted and the correlation function is retrieved from the sequence of the photon arrival times. Single auto- and cross-correlation functions can be processed online in full real time up to count rates of ˜1.8 MHz and ˜1.2 MHz, respectively. Two autocorrelation (A-A, B-B) and a cross correlation (A-B) functions can be simultaneously processed in full real time only up to count rates of ˜750 kHz. At higher count rates, the online processing takes place in a delayed modality, but with no data loss. When tested with simulated correlation data and latex spheres solutions, the overall performances of the correlator appear to be comparable with those of commercial hardware correlators, but with several nontrivial advantages related to its flexibility, low cost, and easy adaptability to future developments of PC and data acquisition technology.

  20. Investigation of adhesion and mechanical properties of human glioma cells by single cell force spectroscopy and atomic force microscopy.

    Andolfi, Laura; Bourkoula, Eugenia; Migliorini, Elisa; Palma, Anita; Pucer, Anja; Skrap, Miran; Scoles, Giacinto; Beltrami, Antonio Paolo; Cesselli, Daniela; Lazzarino, Marco

    2014-01-01

    Active cell migration and invasion is a peculiar feature of glioma that makes this tumor able to rapidly infiltrate into the surrounding brain tissue. In our recent work, we identified a novel class of glioma-associated-stem cells (defined as GASC for high-grade glioma--HG--and Gasc for low-grade glioma--LG) that, although not tumorigenic, act supporting the biological aggressiveness of glioma-initiating stem cells (defined as GSC for HG and Gsc for LG) favoring also their motility. Migrating cancer cells undergo considerable molecular and cellular changes by remodeling their cytoskeleton and cell interactions with surrounding environment. To get a better understanding about the role of the glioma-associated-stem cells in tumor progression, cell deformability and interactions between glioma-initiating stem cells and glioma-associated-stem cells were investigated. Adhesion of HG/LG-cancer cells on HG/LG-glioma-associated stem cells was studied by time-lapse microscopy, while cell deformability and cell-cell adhesion strengths were quantified by indentation measurements by atomic force microscopy and single cell force spectroscopy. Our results demonstrate that for both HG and LG glioma, cancer-initiating-stem cells are softer than glioma-associated-stem cells, in agreement with their neoplastic features. The adhesion strength of GSC on GASC appears to be significantly lower than that observed for Gsc on Gasc. Whereas, GSC spread and firmly adhere on Gasc with an adhesion strength increased as compared to that obtained on GASC. These findings highlight that the grade of glioma-associated-stem cells plays an important role in modulating cancer cell adhesion, which could affect glioma cell migration, invasion and thus cancer aggressiveness. Moreover this work provides evidence about the importance of investigating cell adhesion and elasticity for new developments in disease diagnostics and therapeutics.

  1. Quantification of leakage from large unilamellar lipid vesicles by fluorescence correlation spectroscopy

    Kristensen, Kasper; Henriksen, Jonas Rosager; Andresen, Thomas Lars

    2014-01-01

    Fluorescence correlation spectroscopy (FCS) is a powerful experimental technique that in recent years has found numerous applications for studying biological phenomena. In this article, we scrutinize one of these applications, namely, FCS as a technique for studying leakage of fluorescent molecul...

  2. Second Harmonic Correlation Spectroscopy: Theory and Principles for Determining Surface Binding Kinetics.

    Sly, Krystal L; Conboy, John C

    2017-06-01

    A novel application of second harmonic correlation spectroscopy (SHCS) for the direct determination of molecular adsorption and desorption kinetics to a surface is discussed in detail. The surface-specific nature of second harmonic generation (SHG) provides an efficient means to determine the kinetic rates of adsorption and desorption of molecular species to an interface without interference from bulk diffusion, which is a significant limitation of fluorescence correlation spectroscopy (FCS). The underlying principles of SHCS for the determination of surface binding kinetics are presented, including the role of optical coherence and optical heterodyne mixing. These properties of SHCS are extremely advantageous and lead to an increase in the signal-to-noise (S/N) of the correlation data, increasing the sensitivity of the technique. The influence of experimental parameters, including the uniformity of the TEM00 laser beam, the overall photon flux, and collection time are also discussed, and are shown to significantly affect the S/N of the correlation data. Second harmonic correlation spectroscopy is a powerful, surface-specific, and label-free alternative to other correlation spectroscopic methods for examining surface binding kinetics.

  3. Point-point and point-line moving-window correlation spectroscopy and its applications

    Zhou, Qun; Sun, Suqin; Zhan, Daqi; Yu, Zhiwu

    2008-07-01

    In this paper, we present a new extension of generalized two-dimensional (2D) correlation spectroscopy. Two new algorithms, namely point-point (P-P) correlation and point-line (P-L) correlation, have been introduced to do the moving-window 2D correlation (MW2D) analysis. The new method has been applied to a spectral model consisting of two different processes. The results indicate that P-P correlation spectroscopy can unveil the details and re-constitute the entire process, whilst the P-L can provide general feature of the concerned processes. Phase transition behavior of dimyristoylphosphotidylethanolamine (DMPE) has been studied using MW2D correlation spectroscopy. The newly proposed method verifies that the phase transition temperature is 56 °C, same as the result got from a differential scanning calorimeter. To illustrate the new method further, a lysine and lactose mixture has been studied under thermo perturbation. Using the P-P MW2D, the Maillard reaction of the mixture was clearly monitored, which has been very difficult using conventional display of FTIR spectra.

  4. Sequence-dependent unfolding kinetics of DNA hairpins studied by nanopore force spectroscopy

    Renner, Stephan; Bessonov, Andrey; Simmel, Friedrich C; Gerland, Ulrich

    2010-01-01

    Nanopore force spectroscopy is used to study the unzipping kinetics of two DNA hairpin molecules with a 12 base pair long stem containing two contiguous stretches of six GC and six AT base pairs in interchanged order. Even though the thermodynamic stabilities of the two structures are nearly the same, they differ greatly in their unzipping kinetics. When the GC segment has to be broken before the AT segment, the unfolding rate is orders of magnitude smaller than in the opposite case. We also investigated hairpins with stem regions consisting only of AT or GC base pairs. The pure AT hairpins translocate much faster than the other hairpins, whereas the pure GC hairpins translocate on similar timescales to the hairpins with only an initial GC segment. For each hairpin, nanopore force spectroscopy is performed for different loading rates and the resulting unzipping distributions are mathematically transformed to a master curve that yields the unfolding rate as a function of applied voltage. This is compared with a stochastic model of the unfolding process for the two sequences for different voltages. The results can be rationalized in terms of the different natures of the free energy landscapes for the unfolding process.

  5. Study on Angelica and its different extracts by Fourier transform infrared spectroscopy and two-dimensional correlation IR spectroscopy

    Liu, Hong-xia; Sun, Su-qin; Lv, Guang-hua; Chan, Kelvin K. C.

    2006-05-01

    In order to develop a rapid and effective analysis method for studying integrally the main constituents in the medicinal materials and their extracts, discriminating the extracts from different extraction process, comparing the categories of chemical constituents in the different extracts and monitoring the qualities of medicinal materials, we applied Fourier transform infrared spectroscopy (FT-IR) associated with second derivative infrared spectroscopy and two-dimensional correlation infrared spectroscopy (2D-IR) to study the main constituents in traditional Chinese medicine Angelica and its different extracts (extracted by petroleum ether, ethanol and water in turn). The findings indicated that FT-IR spectrum can provide many holistic variation rules of chemical constituents. Use of the macroscopical fingerprint characters of FT-IR and 2D-IR spectrum can not only identify the main chemical constituents in medicinal materials and their different extracts, but also compare the components differences among the similar samples. This analytical method is highly rapid, effective, visual and accurate for pharmaceutical research.

  6. The correlation between surface electromyography and bite force of mastication muscles in Asian young adults.

    Yen, Cheng-I; Mao, Shih-Hsuan; Chen, Chih-Hao; Chen, Chien-Tzung; Lee, Ming-Yih

    2015-05-01

    Mastication function is related to mandible movement, muscle strength, and bite force. No standard device for measuring bite force has been developed. A linear relationship between electromyographic activity and bite force has been reported by several investigators, but data on the reliability of this relationship remain limited in Asian young adults. The purpose of this study was to develop a clinically applicable, reliable, quantitative, and noninvasive system to measure the kinetic mastication function and observe the correlation between surface electromyography (sEMG) and bite force. The study group consisted of 41 young healthy adults (24 men and 17 women). Surface electromyography was used to evaluate bilateral temporalis and masseter muscle activities, and an occlusal bite force system was used concurrently to measure the bite force during maximal voluntary biting. Bilateral symmetry was compared, and the correlation between EMG and bite force was calculated. The sEMG signals were 107.7±55.0 μV and 106.0±56.0 μV (P=0.699) on right and left temporalis muscles and 183.7±86.2 μV and 194.8±94.3 μV (P=0.121) on right and left masseter muscles, respectively. The bite force was 5.0±3.2 kg on the right side and 5.7±4.0 kg on the left side (P=0.974). A positive correlation between sEMG and bite force was observed. The correlation coefficient between the temporalis muscle and bite force was 0.512, and that between the masseter muscle and bite force was 0.360. No significant difference between the bilateral electromyographic activities of the temporalis and masseter muscles and bilateral bite force was observed in young healthy adults in Taiwan. A positive correlation between sEMG signals and bite force was noted. By combining sEMG and bite force, we developed a clinically applicable, quantitative, reliable, and noninvasive system for evaluating mastication function by using characteristics of biofeedback.

  7. SO2 EMISSION MEASUREMENT BY DOAS (DIFFERENTIAL OPTICAL ABSORPTION SPECTROSCOPY AND COSPEC (CORRELATION SPECTROSCOPY AT MERAPI VOLCANO (INDONESIA

    Hanik Humaida

    2010-06-01

    Full Text Available The SO2 is one of the volcanic gases that can use as indicator of volcano activity. Commonly, SO2 emission is measured by COSPEC (Correlation Spectroscopy. This equipment has several disadvantages; such as heavy, big in size, difficulty in finding spare part, and expensive. DOAS (Differential Optical Absorption Spectroscopy is a new method for SO2 emission measurement that has advantages compares to the COSPEC. Recently, this method has been developed. The SO2 gas emission measurement of Gunung Merapi by DOAS has been carried out at Kaliadem, and also by COSPEC method as comparation. The differences of the measurement result of both methods are not significant. However, the differences of minimum and maximum result of DOAS method are smaller than that of the COSPEC. It has range between 51 ton/day and 87 ton/day for DOAS and 87 ton/day and 201 ton/day for COSPEC. The measurement of SO2 gas emission evaluated with the seismicity data especially the rockfall showed the presence of the positive correlation. It may cause the gas pressure in the subsurface influencing instability of 2006 eruption lava.   Keywords: SO2 gas, Merapi, DOAS, COSPEC

  8. Correlative Raman spectroscopy and focused ion beam for targeted phase boundary analysis of titania polymorphs

    Mangum, John S.; Chan, Lisa H.; Schmidt, Ute; Garten, Lauren M.; Ginley, David S.; Gorman, Brian P.

    2018-05-01

    Site-specific preparation of specimens using focused ion beam instruments for transmission electron microscopy is at the forefront of targeting regions of interest for nanoscale characterization. Typical methods of pinpointing desired features include electron backscatter diffraction for differentiating crystal structures and energy-dispersive X-Ray spectroscopy for probing compositional variations. Yet there are situations, notably in the titanium dioxide system, where these techniques can fail. Differentiating between the brookite and anatase polymorphs of titania is either excessively laborious or impossible with the aforementioned techniques. However, due to differences in bonding structure, Raman spectroscopy serves as an ideal candidate for polymorph differentiation. In this work, a correlative approach utilizing Raman spectroscopy for targeted focused ion beam specimen preparation was employed. Dark field imaging and diffraction in the transmission electron microscope confirmed the region of interest located via Raman spectroscopy and demonstrated the validity of this new method. Correlative Raman spectroscopy, scanning electron microscopy, and focused ion beam is shown to be a promising new technique for identifying site-specific preparation of nanoscale specimens in cases where conventional approaches do not suffice.

  9. Determining the static electronic and vibrational energy correlations via two-dimensional electronic-vibrational spectroscopy

    Dong, Hui; Lewis, Nicholas H. C.; Oliver, Thomas A. A.; Fleming, Graham R., E-mail: grfleming@lbl.gov [Department of Chemistry, University of California, Berkeley, California 94720 (United States); Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, Californial 94720 (United States); Kavli Energy NanoSciences Institute at Berkeley, Berkeley, California 94720 (United States)

    2015-05-07

    Changes in the electronic structure of pigments in protein environments and of polar molecules in solution inevitably induce a re-adaption of molecular nuclear structure. Both changes of electronic and vibrational energies can be probed with visible or infrared lasers, such as two-dimensional electronic spectroscopy or vibrational spectroscopy. The extent to which the two changes are correlated remains elusive. The recent demonstration of two-dimensional electronic-vibrational (2DEV) spectroscopy potentially enables a direct measurement of this correlation experimentally. However, it has hitherto been unclear how to characterize the correlation from the spectra. In this paper, we present a theoretical formalism to demonstrate the slope of the nodal line between the excited state absorption and ground state bleach peaks in the spectra as a characterization of the correlation between electronic and vibrational transition energies. We also show the dynamics of the nodal line slope is correlated to the vibrational spectral dynamics. Additionally, we demonstrate the fundamental 2DEV spectral line-shape of a monomer with newly developed response functions.

  10. Two-dimensional NMR spectroscopy: correlated, homonuclear-correlated, and nuclear Overhauser spectroscopy. January 1975-December 1988 (Citations from the INSPEC: Information Services for the Physics and Engineering Communities data base). Report for January 1975-December 1988

    1988-12-01

    This bibliography contains citations concerning the enhanced analytical techniques of two-dimensional nuclear magnetic resonance (2-D NMR). Applications to specific molecules, biomolecules, and compounds as well as comparisons of three 2-D NMR techniques: correlated spectroscopy (COSY), nuclear Overhauser (NOSEY), and homonuclear-correlated spectroscopy (HOMCOR). (Contains 190 citations fully indexed and including a title list.)

  11. Resolving dual binding conformations of cellulosome cohesin-dockerin complexes using single-molecule force spectroscopy.

    Jobst, Markus A; Milles, Lukas F; Schoeler, Constantin; Ott, Wolfgang; Fried, Daniel B; Bayer, Edward A; Gaub, Hermann E; Nash, Michael A

    2015-10-31

    Receptor-ligand pairs are ordinarily thought to interact through a lock and key mechanism, where a unique molecular conformation is formed upon binding. Contrary to this paradigm, cellulosomal cohesin-dockerin (Coh-Doc) pairs are believed to interact through redundant dual binding modes consisting of two distinct conformations. Here, we combined site-directed mutagenesis and single-molecule force spectroscopy (SMFS) to study the unbinding of Coh:Doc complexes under force. We designed Doc mutations to knock out each binding mode, and compared their single-molecule unfolding patterns as they were dissociated from Coh using an atomic force microscope (AFM) cantilever. Although average bulk measurements were unable to resolve the differences in Doc binding modes due to the similarity of the interactions, with a single-molecule method we were able to discriminate the two modes based on distinct differences in their mechanical properties. We conclude that under native conditions wild-type Doc from Clostridium thermocellum exocellulase Cel48S populates both binding modes with similar probabilities. Given the vast number of Doc domains with predicted dual binding modes across multiple bacterial species, our approach opens up new possibilities for understanding assembly and catalytic properties of a broad range of multi-enzyme complexes.

  12. Reconsideration of dynamic force spectroscopy analysis of streptavidin-biotin interactions.

    Taninaka, Atsushi; Takeuchi, Osamu; Shigekawa, Hidemi

    2010-05-13

    To understand and design molecular functions on the basis of molecular recognition processes, the microscopic probing of the energy landscapes of individual interactions in a molecular complex and their dependence on the surrounding conditions is of great importance. Dynamic force spectroscopy (DFS) is a technique that enables us to study the interaction between molecules at the single-molecule level. However, the obtained results differ among previous studies, which is considered to be caused by the differences in the measurement conditions. We have developed an atomic force microscopy technique that enables the precise analysis of molecular interactions on the basis of DFS. After verifying the performance of this technique, we carried out measurements to determine the landscapes of streptavidin-biotin interactions. The obtained results showed good agreement with theoretical predictions. Lifetimes were also well analyzed. Using a combination of cross-linkers and the atomic force microscope that we developed, site-selective measurement was carried out, and the steps involved in bonding due to microscopic interactions are discussed using the results obtained by site-selective analysis.

  13. Single molecule force spectroscopy data and BD- and MD simulations on the blood protein von Willebrand factor

    Sandra Posch

    2016-09-01

    Full Text Available We here give information for a deeper understanding of single molecule force spectroscopy (SMFS data through the example of the blood protein von Willebrand factor (VWF. It is also shown, how fitting of rupture forces versus loading rate profiles in the molecular dynamics (MD loading-rate range can be used to demonstrate the qualitative agreement between SMFS and MD simulations. The recently developed model by Bullerjahn, Sturm, and Kroy (BSK was used for this demonstration. Further, Brownian dynamics (BD simulations, which can be utilized to estimate the lifetimes of intramolecular VWF interactions under physiological shear, are described. For interpretation and discussion of the methods and data presented here, we would like to directly point the reader to the related research paper, “Mutual A domain interactions in the force sensing protein von Willebrand Factor” (Posch et al., 2016 [1]. Keywords: Atomic force microscopy, Single molecule force spectroscopy, Molecular dynamics simulation, Brownian dynamics simulation, von Willebrand factor

  14. Analysis of dispersive interactions at polymer/TiAlN interfaces by means of dynamic force spectroscopy.

    Wiesing, M; de Los Arcos, T; Gebhard, M; Devi, A; Grundmeier, G

    2017-12-20

    The structural and electronic origins of the interactions between polycarbonate and sputter deposited TiAlN were analysed using a combined electron and force spectroscopic approach. Interaction forces were measured by means of dynamic force spectroscopy and the surface polarizability was analysed by X-ray photoelectron valence band spectroscopy. It could be shown that the adhesive interactions between polycarbonate and TiAlN are governed by van der Waals forces. Different surface cleansing and oxidizing treatments were investigated and the effect of the surface chemistry on the force interactions was analysed. Intense surface oxidation resulted in a decreased adhesion force by a factor of two due to the formation of a 2 nm thick Ti 0.21 Al 0.45 O surface oxide layer. The origin of the residual adhesion forces caused by the mixed Ti 0.21 Al 0.45 O surface oxide was clarified by considering the non-retarded Hamaker coefficients as calculated by Lifshitz theory, based on optical data from Reflection Electron Energy Loss Spectroscopy. This disclosed increased dispersion forces of Ti 0.21 Al 0.45 O due to the presence of Ti(iv) ions and related Ti 3d band optical transitions.

  15. [Classification of results of studying blood plasma with laser correlation spectroscopy based on semiotics of preclinical and clinical states].

    Ternovoĭ, K S; Kryzhanovskiĭ, G N; Musiĭchuk, Iu I; Noskin, L A; Klopov, N V; Noskin, V A; Starodub, N F

    1998-01-01

    The usage of laser correlation spectroscopy for verification of preclinical and clinical states is substantiated. Developed "semiotic" classifier for solving the problems of preclinical and clinical states is presented. The substantiation of biological algorithms as well as the mathematical support and software for the proposed classifier for the data of laser correlation spectroscopy of blood plasma are presented.

  16. Correlation between near infrared spectroscopy and electrical techniques in measuring skin moisture content

    Mohamad, M; Sabbri, A R M; Jafri, M Z Mat; Omar, A F

    2014-01-01

    Near infrared (NIR) spectroscopy technique serves as an important tool for the measurement of moisture content of skin owing to the advantages it has over the other techniques. The purpose of the study is to develop a correlation between NIR spectrometer with electrical conventional techniques for skin moisture measurement. A non-invasive measurement of moisture content of skin was performed on different part of human face and hand under control environment (temperature 21 ± 1 °C, relative humidity 45 ± 5 %). Ten healthy volunteers age between 21-25 (male and female) participated in this study. The moisture content of skin was measured using DermaLab ® USB Moisture Module, Scalar Moisture Checker and NIR spectroscopy (NIRQuest). Higher correlation was observed between NIRQuest and Dermalab moisture probe with a coefficient of determination (R 2 ) above 70 % for all the subjects. However, the value of R 2 between NIRQuest and Moisture Checker was observed to be lower with the R 2 values ranges from 51.6 to 94.4 %. The correlation of NIR spectroscopy technique successfully developed for measuring moisture content of the skin. The analysis of this correlation can help to establish novel instruments based on an optical system in clinical used especially in the dermatology field

  17. The Pharmaceutical Capping Process-Correlation between Residual Seal Force, Torque Moment, and Flip-off Removal Force.

    Mathaes, Roman; Mahler, Hanns-Christian; Vorgrimler, Lothar; Steinberg, Henrik; Dreher, Sascha; Roggo, Yves; Nieto, Alejandra; Brown, Helen; Roehl, Holger; Adler, Michael; Luemkemann, Joerg; Huwyler, Joerg; Lam, Philippe; Stauch, Oliver; Mohl, Silke; Streubel, Alexander

    2016-01-01

    The majority of parenteral drug products are manufactured in glass vials with an elastomeric rubber stopper and a crimp cap. The vial sealing process is a critical process step during fill-and-finish operations, as it defines the seal quality of the final product. Different critical capping process parameters can affect rubber stopper defects, rubber stopper compression, container closure integrity, and also crimp cap quality. A sufficiently high force to remove the flip-off button prior to usage is required to ensure quality of the drug product unit by the flip-off button during storage, transportation, and until opening and use. Therefore, the final product is 100% visually inspected for lose or defective crimp caps, which is subjective as well as time- and labor-intensive. In this study, we sealed several container closure system configurations with different capping equipment settings (with corresponding residual seal force values) to investigate the torque moment required to turn the crimp cap. A correlation between torque moment and residual seal force has been established. The torque moment was found to be influenced by several parameters, including diameter of the vial head, type of rubber stopper (serum or lyophilized) and type of crimp cap (West(®) or Datwyler(®)). In addition, we measured the force required to remove the flip-off button of a sealed container closure system. The capping process had no influence on measured forces; however, it was possible to detect partially crimped vials. In conclusion, a controlled capping process with a defined target residual seal force range leads to a tight crimp cap on a sealed container closure system and can ensure product quality. The majority of parenteral drug products are manufactured in a glass vials with an elastomeric rubber stopper and a crimp cap. The vial sealing process is a critical process step during fill-and-finish operations, as it defines the seal quality of the final product. An adequate force

  18. Improving estimation of kinetic parameters in dynamic force spectroscopy using cluster analysis

    Yen, Chi-Fu; Sivasankar, Sanjeevi

    2018-03-01

    Dynamic Force Spectroscopy (DFS) is a widely used technique to characterize the dissociation kinetics and interaction energy landscape of receptor-ligand complexes with single-molecule resolution. In an Atomic Force Microscope (AFM)-based DFS experiment, receptor-ligand complexes, sandwiched between an AFM tip and substrate, are ruptured at different stress rates by varying the speed at which the AFM-tip and substrate are pulled away from each other. The rupture events are grouped according to their pulling speeds, and the mean force and loading rate of each group are calculated. These data are subsequently fit to established models, and energy landscape parameters such as the intrinsic off-rate (koff) and the width of the potential energy barrier (xβ) are extracted. However, due to large uncertainties in determining mean forces and loading rates of the groups, errors in the estimated koff and xβ can be substantial. Here, we demonstrate that the accuracy of fitted parameters in a DFS experiment can be dramatically improved by sorting rupture events into groups using cluster analysis instead of sorting them according to their pulling speeds. We test different clustering algorithms including Gaussian mixture, logistic regression, and K-means clustering, under conditions that closely mimic DFS experiments. Using Monte Carlo simulations, we benchmark the performance of these clustering algorithms over a wide range of koff and xβ, under different levels of thermal noise, and as a function of both the number of unbinding events and the number of pulling speeds. Our results demonstrate that cluster analysis, particularly K-means clustering, is very effective in improving the accuracy of parameter estimation, particularly when the number of unbinding events are limited and not well separated into distinct groups. Cluster analysis is easy to implement, and our performance benchmarks serve as a guide in choosing an appropriate method for DFS data analysis.

  19. Single Cell Force Spectroscopy for Quantification of Cellular Adhesion on Surfaces

    Christenson, Wayne B.

    Cell adhesion is an important aspect of many biological processes. The atomic force microscope (AFM) has made it possible to quantify the forces involved in cellular adhesion using a technique called single cell force spectroscopy (SCFS). AFM based SCFS offers versatile control over experimental conditions for probing directly the interaction between specific cell types and specific proteins, surfaces, or other cells. Transmembrane integrins are the primary proteins involved in cellular adhesion to the extra cellular matix (ECM). One of the chief integrins involved in the adhesion of leukocyte cells is alpha Mbeta2 (Mac-1). The experiments in this dissertation quantify the adhesion of Mac-1 expressing human embryonic kidney (HEK Mac-1), platelets, and neutrophils cells on substrates with different concentrations of fibrinogen and on fibrin gels and multi-layered fibrinogen coated fibrin gels. It was shown that multi-layered fibrinogen reduces the adhesion force of these cells considerably. A novel method was developed as part of this research combining total internal reflection microscopy (TIRFM) with SCFS allowing for optical microscopy of HEK Mac-1 cells interacting with bovine serum albumin (BSA) coated glass after interacting with multi-layered fibrinogen. HEK Mac-1 cells are able to remove fibrinogen molecules from the multi-layered fibrinogen matrix. An analysis methodology for quantifying the kinetic parameters of integrin-ligand interactions from SCFS experiments is proposed, and the kinetic parameters of the Mac-1 fibrinogen bond are quantified. Additional SCFS experiments quantify the adhesion of macrophages and HEK Mac-1 cells on functionalized glass surfaces and normal glass surfaces. Both cell types show highest adhesion on a novel functionalized glass surface that was prepared to induce macrophage fusion. These experiments demonstrate the versatility of AFM based SCFS, and how it can be applied to address many questions in cellular biology offering

  20. Spatio-Temporal Image Correlation Spectroscopy Measurements of Flow Demonstrated in Microfluidic Channels

    Rossow, Molly; Mantulin, William W.; Gratton, Enrico

    2009-01-01

    Accurate blood flow measurements during surgery can improve the operations chance of success. We developed Near-infrared Spatio-Temporal Image Spectroscopy (NIR-STICS), which has the potential to make blood flow measurements that are difficult to accomplish with existing methods. Specifically, we propose the technique and we show feasibility on phantom measurements. NIR-STICS has the potential of measuring the fluid velocity in small blood vessels (less than 1mm in diameter) and of creating a map of blood flow rates over an area of approximately 1cm2. NIR-STICS employs near-infrared spectroscopy to probe inside blood vessel walls and spatio-temporal image correlation spectroscopy to directly—without the use of a model—extract fluid velocity from the fluctuations within an image. Here we present computer simulations and experiments on a phantom system that demonstrate the effectiveness of NIR-STICS. PMID:19405744

  1. Spatiotemporal image correlation spectroscopy measurements of flow demonstrated in microfluidic channels

    Rossow, Molly; Mantulin, William W.; Gratton, Enrico

    2009-03-01

    Accurate blood flow measurements during surgery can improve an operation's chance of success. We developed near-infrared spatio-temporal image spectroscopy (NIR-STICS), which has the potential to make blood flow measurements that are difficult to accomplish with existing methods. Specifically, we propose the technique and we show feasibility on phantom measurements. NIR-STICS has the potential of measuring the fluid velocity in small blood vessels (less than 1 mm in diameter) and of creating a map of blood flow rates over an area of approximately 1 cm2. NIR-STICS employs near-infrared spectroscopy to probe inside blood vessel walls and spatiotemporal image correlation spectroscopy to directly-without the use of a model-extract fluid velocity from the fluctuations within an image. We present computer simulations and experiments on a phantom system that demonstrate the effectiveness of NIR-STICS.

  2. Sensitivity of near-infrared spectroscopy and diffuse correlation spectroscopy to brain hemodynamics: simulations and experimental findings during hypercapnia

    Selb, Juliette; Boas, David A.; Chan, Suk-Tak; Evans, Karleyton C.; Buckley, Erin M.; Carp, Stefan A.

    2014-01-01

    Abstract. Near-infrared spectroscopy (NIRS) and diffuse correlation spectroscopy (DCS) are two diffuse optical technologies for brain imaging that are sensitive to changes in hemoglobin concentrations and blood flow, respectively. Measurements for both modalities are acquired on the scalp, and therefore hemodynamic processes in the extracerebral vasculature confound the interpretation of cortical hemodynamic signals. The sensitivity of NIRS to the brain versus the extracerebral tissue and the contrast-to-noise ratio (CNR) of NIRS to cerebral hemodynamic responses have been well characterized, but the same has not been evaluated for DCS. This is important to assess in order to understand their relative capabilities in measuring cerebral physiological changes. We present Monte Carlo simulations on a head model that demonstrate that the relative brain-to-scalp sensitivity is about three times higher for DCS (0.3 at 3 cm) than for NIRS (0.1 at 3 cm). However, because DCS has higher levels of noise due to photon-counting detection, the CNR is similar for both modalities in response to a physiologically realistic simulation of brain activation. Even so, we also observed higher CNR of the hemodynamic response during graded hypercapnia in adult subjects with DCS than with NIRS. PMID:25453036

  3. Sensitivity of near-infrared spectroscopy and diffuse correlation spectroscopy to brain hemodynamics: simulations and experimental findings during hypercapnia.

    Selb, Juliette; Boas, David A; Chan, Suk-Tak; Evans, Karleyton C; Buckley, Erin M; Carp, Stefan A

    2014-07-01

    Near-infrared spectroscopy (NIRS) and diffuse correlation spectroscopy (DCS) are two diffuse optical technologies for brain imaging that are sensitive to changes in hemoglobin concentrations and blood flow, respectively. Measurements for both modalities are acquired on the scalp, and therefore hemodynamic processes in the extracerebral vasculature confound the interpretation of cortical hemodynamic signals. The sensitivity of NIRS to the brain versus the extracerebral tissue and the contrast-to-noise ratio (CNR) of NIRS to cerebral hemodynamic responses have been well characterized, but the same has not been evaluated for DCS. This is important to assess in order to understand their relative capabilities in measuring cerebral physiological changes. We present Monte Carlo simulations on a head model that demonstrate that the relative brain-to-scalp sensitivity is about three times higher for DCS (0.3 at 3 cm) than for NIRS (0.1 at 3 cm). However, because DCS has higher levels of noise due to photon-counting detection, the CNR is similar for both modalities in response to a physiologically realistic simulation of brain activation. Even so, we also observed higher CNR of the hemodynamic response during graded hypercapnia in adult subjects with DCS than with NIRS.

  4. Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy

    Laňková, Martina; Humpolíčková, Jana; Vosolsobě, S.; Cit, Zdeněk; Lacek, Jozef; Čovan, Martin; Čovanová, Milada; Hof, Martin; Petrášek, Jan

    2016-01-01

    Roč. 22, č. 2 (2016), s. 290-299 ISSN 1431-9276 R&D Projects: GA ČR(CZ) GAP305/11/2476; GA ČR(CZ) GPP501/12/P951 Institutional support: RVO:61389030 ; RVO:61388955 Keywords : raster image correlation spectroscopy * fluorescence recovery after photobleaching * auxin influx Subject RIV: EB - Genetics ; Molecular Biology; CF - Physical ; Theoretical Chemistry (UFCH-W) Impact factor: 1.891, year: 2016

  5. Insight into Resolution Enhancement in Generalized Two-Dimensional Correlation Spectroscopy

    Ma, Lu; Sikirzhytski, Vitali; Hong, Zhenmin; Lednev, Igor K.; Asher, Sanford A.

    2013-01-01

    Generalized two-dimensional correlation spectroscopy (2D COS) can be used to enhance spectral resolution in order to help differentiate highly overlapped spectral bands. Despite the numerous extensive 2D COS investigations, the origin of the 2D spectral resolution enhancement mechanism(s) are not completely understood. In the work here we studied the 2D COS of simulated spectra in order to develop new insights into the dependence of the 2D COS spectral features on the overlapping band separat...

  6. Quantification of transuranic elements by time interval correlation spectroscopy of the detected neutrons

    Baeten; Bruggeman; Paepen; Carchon

    2000-03-01

    The non-destructive quantification of transuranic elements in nuclear waste management or in safeguards verifications is commonly performed by passive neutron assay techniques. To minimise the number of unknown sample-dependent parameters, Neutron Multiplicity Counting (NMC) is applied. We developed a new NMC-technique, called Time Interval Correlation Spectroscopy (TICS), which is based on the measurement of Rossi-alpha time interval distributions. Compared to other NMC-techniques, TICS offers several advantages.

  7. The use of photon correlation spectroscopy method for determining diffusion coefficient in brine and herring flesh

    Shumanova M.V.

    2015-03-01

    Full Text Available The process fish salting has been studied by the method of photon correlation spectroscopy; the distribution of salt concentration in the solution and herring flesh with skin has been found, diffusion coefficients and salt concentrations used for creating a mathematical model of the salting technology have been worked out; the possibility of determination by this method the coefficient of dynamic viscosity of solutions and different media (minced meat etc. has been considered

  8. Data acquisition card for fluctuation correlation spectroscopy allowing full access to the detected photon sequence

    Eid, JS; Muller, JD; Gratton, E

    2000-01-01

    Typically, fluctuation correlation spectroscopy (FCS) data acquisition cards measure the number of photon events per time interval (i.e., bin) - time mode. Commercial FCS cards combine the bins through hardware in order to calculate the autocorrelation function. Such a design therefore does not yield the time resolved photon sequence, but only the autocorrelation of that sequence. A different acquisition method which measures the number of time intervals between photon events has been impleme...

  9. Integrin and glycocalyx mediated contributions to cell adhesion identified by single cell force spectroscopy

    Boettiger, D; Wehrle-Haller, B

    2010-01-01

    The measurement of cell adhesion using single cell force spectroscopy methods was compared with earlier methods for measuring cell adhesion. This comparison provided a means and rationale for separating components of the measurement retract curve that were due to interactions between the substrate and the glycocalyx, and interactions that were due to cell surface integrins binding to a substrate-bound ligand. The glycocalyx adhesion was characterized by multiple jumps with dispersed jump sizes that extended from 5 to 30 μm from the origin. The integrin mediated adhesion was represented by the F max (maximum detachment force), was generally within the first 5 μm and commonly detached with a single rupture cascade. The integrin peak (F max ) increases with time and the rate of increase shows large cell to cell variability with a peak ∼ 50 nN s -1 and an average rate of increase of 75 pN s -1 . This is a measure of the rate of increase in the number of adhesive integrin-ligand bonds/cell as a function of contact time.

  10. 2D Vis/NIR correlation spectroscopy of cooked chicken meats

    Liu, Yongliang; Chen, Yud-Ren; Ozaki, Yukihiro

    2000-03-01

    Cooking of chicken meats was investigated by the generalized two-dimensional visible/near-infrared (2D Vis/NIR) correlation spectroscopy. Synchronous and asynchronous spectra in the 400-700 nm visible region suggested that the 445 and 560 nm bands be ascribed to deoxymyoglobin and oxymyoglobin, and at least one of the 475, 520, and 585 nm bands is assignable to the denatured species (metmyoglobin). The asynchronous 2D NIR correlation spectrum showed that CH bands change their spectral intensities before the OH/NH groups during the cooking process, indicating that CH fractions are easily oxidized and degraded. In addition, strong correlation peaks were observed correlating the bands in the visible and NIR spectral regions.

  11. Force Spectroscopy of the Plasmodium falciparum Vaccine Candidate Circumsporozoite Protein Suggests a Mechanically Pliable Repeat Region.

    Patra, Aditya Prasad; Sharma, Shobhona; Ainavarapu, Sri Rama Koti

    2017-02-10

    The most effective vaccine candidate of malaria is based on the Plasmodium falciparum circumsporozoite protein (CSP), a major surface protein implicated in the structural strength, motility, and immune evasion properties of the infective sporozoites. It is suspected that reversible conformational changes of CSP are required for infection of the mammalian host, but the detailed structure and dynamic properties of CSP remain incompletely understood, limiting our understanding of its function in the infection. Here, we report the structural and mechanical properties of the CSP studied using single-molecule force spectroscopy on several constructs, one including the central region of CSP, which is rich in NANP amino acid repeats (CSP rep ), and a second consisting of a near full-length sequence without the signal and anchor hydrophobic domains (CSP ΔHP ). Our results show that the CSP rep is heterogeneous, with 40% of molecules requiring virtually no mechanical force to unfold (<10 piconewtons (pN)), suggesting that these molecules are mechanically compliant and perhaps act as entropic springs, whereas the remaining 60% are partially structured with low mechanical resistance (∼70 pN). CSP ΔHP having multiple force peaks suggests specifically folded domains, with two major populations possibly indicating the open and collapsed forms. Our findings suggest that the overall low mechanical resistance of the repeat region, exposed on the outer surface of the sporozoites, combined with the flexible full-length conformations of CSP, may provide the sporozoites not only with immune evasion properties, but also with lubricating capacity required during its navigation through the mosquito and vertebrate host tissues. We anticipate that these findings would further assist in the design and development of future malarial vaccines. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Force-induced unzipping of DNA with long-range correlated sequence

    Allahverdyan, A. E.; Gevorkian, Zh. S.

    2002-01-01

    We consider force-induced unzipping transition for a heterogeneous DNA model with a long-range correlated base-sequence. It is shown that as compared to the uncorrelated situation, long-range correlations smear the unzipping phase-transition, change its universality class and lead to non-self-averaging: the averaged behavior strongly differs from the typical ones. Several basic scenarios for this typical behavior are revealed and explained. The results can be relevant for explaining the biolo...

  13. Preventing probe induced topography correlated artifacts in Kelvin Probe Force Microscopy

    Polak, L.; Wijngaarden, Rinke J.

    2016-01-01

    Kelvin Probe Force Microscopy (KPFM) on samples with rough surface topography can be hindered by topography correlated artifacts. We show that, with the proper experimental configuration and using homogeneously metal coated probes, we are able to obtain amplitude modulation (AM) KPFM results on a

  14. Femtosecond spectroscopy in semiconductors: a key to coherences, correlations and quantum kinetics

    Axt, V M; Kuhn, T

    2004-01-01

    The application of femtosecond spectroscopy to the study of ultrafast dynamics in semiconductor materials and nanostructures is reviewed with particular emphasis on the physics that can be learned from it. Excitation with ultrashort optical pulses in general results in the creation of coherent superpositions and correlated many-particle states. The review comprises a discussion of the dynamics of this correlated many-body system during and after pulsed excitation as well as its analysis by means of refined measurements and advanced theories. After an introduction of basic concepts-such as coherence, correlation and quantum kinetics-a brief overview of the most important experimental techniques and theoretical approaches is given. The remainder of this paper is devoted to specific results selected in order to highlight how femtosecond spectroscopy gives access to the physics of coherences, correlations and quantum kinetics involving charge, spin and lattice degrees of freedom. First examples deal with the dynamics of basic laser-induced coherences that can be observed, e.g. in quantum beat spectroscopy, in coherent control measurements or in experiments using few-cycle pulses. The phenomena discussed here are basic in the sense that they can be understood to a large extent on the mean-field level of the theory. Nevertheless, already on this level it is found that semiconductors behave substantially differently from atomic systems. Subsequent sections report on the occurrence of coherences and correlations beyond the mean-field level that are mediated either by carrier-phonon or carrier-carrier interactions. The corresponding analysis gives deep insight into fundamental issues such as the energy-time uncertainty, pure dephasing in quantum dot structures, the role of two-pair or even higher correlations and the build-up of screening. Finally results are presented concerning the ultrafast dynamics of resonantly coupled excitations, where a combination of different

  15. Fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy reveal the cytoplasmic origination of loaded nuclear RISC in vivo in human cells.

    Ohrt, Thomas; Mütze, Jörg; Staroske, Wolfgang; Weinmann, Lasse; Höck, Julia; Crell, Karin; Meister, Gunter; Schwille, Petra

    2008-11-01

    Studies of RNA interference (RNAi) provide evidence that in addition to the well-characterized cytoplasmic mechanisms, nuclear mechanisms also exist. The mechanism by which the nuclear RNA-induced silencing complex (RISC) is formed in mammalian cells, as well as the relationship between the RNA silencing pathways in nuclear and cytoplasmic compartments is still unknown. Here we show by applying fluorescence correlation and cross-correlation spectroscopy (FCS/FCCS) in vivo that two distinct RISC exist: a large approximately 3 MDa complex in the cytoplasm and a 20-fold smaller complex of approximately 158 kDa in the nucleus. We further show that nuclear RISC, consisting only of Ago2 and a short RNA, is loaded in the cytoplasm and imported into the nucleus. The loaded RISC accumulates in the nucleus depending on the presence of a target, based on an miRNA-like interaction with impaired cleavage of the cognate RNA. Together, these results suggest a new RISC shuttling mechanism between nucleus and cytoplasm ensuring concomitant gene regulation by small RNAs in both compartments.

  16. Force spectroscopy of membrane hardness of SH-SY5Y neuroblastoma cells before and after differentiation

    Kwon, Sangwoo; Yang, Woochul; Choi, Yun Kyong; Park, Jung Keuck

    2014-05-01

    Atomic force microscopy (AFM) is utilized in many studies for measuring the structure and the physical characteristics of soft and bio materials. In particular, the force spectroscopy function in the AFM system allows us to explore the mechanical properties of bio cells. In this study, we probe the variation in the membrane hardness of human neuroblastoma SH-SY5Y cells (SH-cells) before and after differentiation by using force spectroscopy. The SH-cell, which is usually differentiated by using a chemical treatment with retinoic acid (RA), is a neuronal cell line employed widely as an in-vitro model for neuroscience research. In force spectroscopy, the force-distance curves are obtained from both the original and the RA-treated cells while the AFM tip approaches and pushes on the cell membranes. The slope deduced from linear region in the force-distance curve is the spring constant and corresponds to the hardness of the cell membrane. The spring constant of the RA-treated cells (0.597 ± 0.010 nN/nm) was smaller than that of the original cells (0.794 ± 0.010 nN/nm), reflecting a hardness decrease in the cells differentiated with the RA treatments. The results clearly demonstrated that the differentiated cells are softer than the original cells. The change in the elasticity of the differentiated cells might be caused by morphological modification during differentiation process. We suggest that force spectroscopy can be employed as a novel method to determine the degree of differentiation of stem cells into various functional cells.

  17. Spectroscopy

    Hellman, Hal

    1968-01-01

    This booklet discusses spectroscopy, the study of absorption of radiation by matter, including X-ray, gamma-ray, microwave, mass spectroscopy, as well as others. Spectroscopy has produced more fundamental information to the study of the detailed structure of matter than any other tools.

  18. Stimulated Raman spectroscopy and nanoscopy of molecules using near field photon induced forces without resonant electronic enhancement gain

    Tamma, Venkata Ananth [CaSTL Center, Department of Chemistry, University of California, Irvine, California 92697 (United States); Huang, Fei; Kumar Wickramasinghe, H., E-mail: hkwick@uci.edu [Department of Electrical Engineering and Computer Science, 142 Engineering Tower, University of California, Irvine, California 92697 (United States); Nowak, Derek [Molecular Vista, Inc., 6840 Via Del Oro, San Jose, California 95119 (United States)

    2016-06-06

    We report on stimulated Raman spectroscopy and nanoscopy of molecules, excited without resonant electronic enhancement gain, and recorded using near field photon induced forces. Photon-induced interaction forces between the sharp metal coated silicon tip of an Atomic Force Microscope (AFM) and a sample resulting from stimulated Raman excitation were detected. We controlled the tip to sample spacing using the higher order flexural eigenmodes of the AFM cantilever, enabling the tip to come very close to the sample. As a result, the detection sensitivity was increased compared with previous work on Raman force microscopy. Raman vibrational spectra of azobenzene thiol and l-phenylalanine were measured and found to agree well with published results. Near-field force detection eliminates the need for far-field optical spectrometer detection. Recorded images show spatial resolution far below the optical diffraction limit. Further optimization and use of ultrafast pulsed lasers could push the detection sensitivity towards the single molecule limit.

  19. Characterization of polymer surface structure and surface mechanical behaviour by sum frequency generation surface vibrational spectroscopy and atomic force microscopy

    Opdahl, Aric; Koffas, Telly S; Amitay-Sadovsky, Ella; Kim, Joonyeong; Somorjai, Gabor A

    2004-01-01

    Sum frequency generation (SFG) vibrational spectroscopy and atomic force microscopy (AFM) have been used to study polymer surface structure and surface mechanical behaviour, specifically to study the relationships between the surface properties of polymers and their bulk compositions and the environment to which the polymer is exposed. The combination of SFG surface vibrational spectroscopy and AFM has been used to study surface segregation behaviour of polyolefin blends at the polymer/air and polymer/solid interfaces. SFG surface vibrational spectroscopy and AFM experiments have also been performed to characterize the properties of polymer/liquid and polymer/polymer interfaces, focusing on hydrogel materials. A method was developed to study the surface properties of hydrogel contact lens materials at various hydration conditions. Finally, the effect of mechanical stretching on the surface composition and surface mechanical behaviour of phase-separated polyurethanes, used in biomedical implant devices, has been studied by both SFG surface vibrational spectroscopy and AFM. (topical review)

  20. Nanostructure and force spectroscopy analysis of human peripheral blood CD4+ T cells using atomic force microscopy

    Hu Mingqian; Wang Jiongkun; Cai Jiye; Wu Yangzhe; Wang Xiaoping

    2008-01-01

    To date, nanoscale imaging of the morphological changes and adhesion force of CD4 + T cells during in vitro activation remains largely unreported. In this study, we used atomic force microscopy (AFM) to study the morphological changes and specific binding forces in resting and activated human peripheral blood CD4 + T cells. The AFM images revealed that the volume of activated CD4 + T cells increased and the ultrastructure of these cells also became complex. Using a functionalized AFM tip, the strength of the specific binding force of the CD4 antigen-antibody interaction was found to be approximately three times that of the unspecific force. The adhesion forces were not randomly distributed over the surface of a single activated CD4 + T cell, indicated that the CD4 molecules concentrated into nanodomains. The magnitude of the adhesion force of the CD4 antigen-antibody interaction did not change markedly with the activation time. Multiple bonds involved in the CD4 antigen-antibody interaction were measured at different activation times. These results suggest that the adhesion force involved in the CD4 antigen-antibody interaction is highly selective and of high affinity

  1. Nanostructure and force spectroscopy analysis of human peripheral blood CD4+ T cells using atomic force microscopy.

    Hu, Mingqian; Wang, Jiongkun; Cai, Jiye; Wu, Yangzhe; Wang, Xiaoping

    2008-09-12

    To date, nanoscale imaging of the morphological changes and adhesion force of CD4(+) T cells during in vitro activation remains largely unreported. In this study, we used atomic force microscopy (AFM) to study the morphological changes and specific binding forces in resting and activated human peripheral blood CD4(+) T cells. The AFM images revealed that the volume of activated CD4(+) T cells increased and the ultrastructure of these cells also became complex. Using a functionalized AFM tip, the strength of the specific binding force of the CD4 antigen-antibody interaction was found to be approximately three times that of the unspecific force. The adhesion forces were not randomly distributed over the surface of a single activated CD4(+) T cell, indicated that the CD4 molecules concentrated into nanodomains. The magnitude of the adhesion force of the CD4 antigen-antibody interaction did not change markedly with the activation time. Multiple bonds involved in the CD4 antigen-antibody interaction were measured at different activation times. These results suggest that the adhesion force involved in the CD4 antigen-antibody interaction is highly selective and of high affinity.

  2. Spectroscopy

    Walker, S

    1976-01-01

    The three volumes of Spectroscopy constitute the one comprehensive text available on the principles, practice and applications of spectroscopy. By giving full accounts of those spectroscopic techniques only recently introduced into student courses - such as Mössbauer spectroscopy and photoelectron spectroscopy - in addition to those techniques long recognised as being essential in chemistry teaching - sucha as e.s.r. and infrared spectroscopy - the book caters for the complete requirements of undergraduate students and at the same time provides a sound introduction to special topics for graduate students.

  3. Slow dynamics in an azopolymer molecular layer studied by x-ray photon correlation spectroscopy

    Orsi, D.; Fluerasu, A.; Cristofolini, L.; Fontana, M.P.; Pontecorvo, E.; Caronna, C.; Zontone, F.; Madsen, A.

    2010-01-01

    We report the results of x-ray photon correlation spectroscopy (XPCS) experiments on multilayers of a photosensitive azo-polymer which can be softened by photoisomerization. Time correlation functions have been measured at different temperatures and momentum transfers (q) and under different illumination conditions (dark, UV or visible). The correlation functions are well described by the Kohlrausch-Williams-Watts (KWW) form with relaxation times that are proportional to q -1 . The characteristic relaxation times follow the same Vogel-Fulcher-Tammann law describing the bulk viscosity of this polymer. The out-of-equilibrium relaxation dynamics following a UV photoperturbation are accelerated, which is in agreement with a fluidification effect previously measured by rheology. The transient dynamics are characterized by two times correlation function, and dynamical heterogeneity is evidenced by calculating the variance χ of the degree of correlation as a function of ageing time. A clear peak in χ appears at a well defined time τ C which scales with q -1 and with the ageing time, in a similar fashion as previously reported in colloidal suspensions (O. Dauchot et al. Phys. Rev. Lett. 95 265701 (2005)). From an accurate analysis of the correlation functions we could demonstrate a temperature and light dependent cross-over from compressed KWW to simple exponential behavior.

  4. The Utilization of Spin Polarized Photoelectron Spectroscopy as a Probe of Electron Correlation with an Ultimate Goal of Pu

    Tobin, James; Yu, Sung; Chung, Brandon; Morton, Simon; Komesu, Takashi; Waddill, George

    2008-01-01

    We are developing the technique of spin-polarized photoelectron spectroscopy as a probe of electron correlation with the ultimate goal of resolving the Pu electronic structure controversy. Over the last several years, we have demonstrated the utility of spin polarized photoelectron spectroscopy for determining the fine details of the electronic structure in complex systems such as those shown in the paper.

  5. Correlation of proton MR spectroscopy of primary intracranial tumours and histopathological findings: preliminarily findings

    Mandel, C.; Birchall, D.; Ellison, D.; Crawford, P.J.

    2002-01-01

    consistently raised in tumours, with elevated Cho:Crnormal and Cho:Chnormal constant findings. Preliminary analysis indicates that the tumoral Cho:Cr and Cho:NAA ratios demonstrate the closest correlation to histological grade. Full statistical analysis will be presented. Proton MR spectroscopy is a useful adjunct to the investigation of primary intracranial tumours. It is in a relatively early phase of clinical validation, but has the potential for use as a form of non-invasive biopsy in selected patients. Copyright (2002) Blackwell Science Pty Ltd

  6. MDM2-MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance.

    Moscetti, Ilaria; Teveroni, Emanuela; Moretti, Fabiola; Bizzarri, Anna Rita; Cannistraro, Salvatore

    Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2-MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2-MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD ) in the micromolar range for the MDM2-MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2-MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2-MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation.

  7. Substrate binding to SGLT1 investigated by single molecule force spectroscopy

    Neundlinger, I. J.

    2010-01-01

    D-glucose serves as one of the most important fuels in various organism due to its fundamental role in ATP-, protein and lipid synthesis. Thus, sustaining glucose homeostasis is a crucial issue of life as disorders can cause severe malfunctions such as glucose-galactose-malabsorbtion (GGM). Sodium-glucose co-transporter, SGLTs, especially the high affinity transporter SGLT1, play a crucial role in accumulation of glucose in the cell as they facilitate transport of the sugar into the cytoplasma across the cell membrane by a Na+-electrochemical potential. Even recently, members of the SGLT transporter family have become a therapeutic target for the treatment of hyperglycemia in type 2 diabetes. Hence, it is of particular importance to gain insights on the dynamic behavior of SGLTs during substrate binding and transport across the cell membrane on the single molecular level. In the present study, the Atomic Force Microscope (AFM) was employed to investigate the dynamic properties of the sodium-glucose co-transporter SGLT1 upon substrate binding under nearly physiological conditions. Hereto, new glucose derivatives were synthesized in order to probe the recognition efficiency of these molecules to SGLT1 embedded in the plasma membrane of living cells. A well established coupling protocol was used to covalently link (i) amino-modified D-glucose owning a conserved pyranose ring, (ii) 1-thio-β-D-glucose having a sulphur atom at C1 of the pyranose ring and (iii) the competitive inhibitor phlorizin to the AFM tip via poly(ethylene)glycol (PEG)-tether using different functional end groups and varying lengths. Binding characteristics, e.g. binding probability, interaction forces, influence of substances (glucose, phlorizin, sodium) and of molecule-linker compounds were obtained by performing single molecular recognition force spectroscopy (SMRFS) measurements. Moreover, temperature controlled radioactive binding/transport assays and SMRFS experiments yielded insights into

  8. Absolute choline concentration measured by quantitative proton MR spectroscopy correlates with cell density in meningioma

    Yue, Qiang [University of Tsukuba, Department of Neurosurgery, Institute of Clinical Medicine, Tsukuba Science City, Ibaraki (Japan)]|[West China Hospital of Sichuan University, Huaxi MR Research Center, Department of Radiology, Chengdu (China); Shibata, Yasushi; Kawamura, Hiraku; Matsumura, Akira [University of Tsukuba, Department of Neurosurgery, Institute of Clinical Medicine, Tsukuba Science City, Ibaraki (Japan); Isobe, Tomonori [Kitasato University, Department of Medical Technology, School of Allied Health Sciences, Minato, Tokyo (Japan); Anno, Izumi [University of Tsukuba, Department of Radiology, Institute of Clinical Medicine, Tsukuba, Ibaraki (Japan); Gong, Qi-Yong [West China Hospital of Sichuan University, Huaxi MR Research Center, Department of Radiology, Chengdu (China)]|[University of Liverpool, Division of Medical Imaging, Faculty of Medicine, Liverpool (United Kingdom)

    2009-01-15

    This study was aimed to investigate the relationship between quantitative proton magnetic resonance spectroscopy (1H-MRS) and pathological changes in meningioma. Twenty-two meningioma cases underwent single voxel 1H-MRS (point-resolved spectroscopy sequence, repetition time/echo time = 2,000 ms/68, 136, 272 ms). Absolute choline (Cho) concentration was calculated using tissue water as the internal reference and corrected according to intra-voxel cystic/necrotic parts. Pathological specimens were stained with MIB-1 antibody to measure cell density and proliferation index. Correlation analysis was performed between absolute Cho concentration and cell density and MIB-1 labeled proliferation index. Average Cho concentration of all meningiomas before correction was 2.95 {+-} 0.86 mmol/kg wet weight. It was increased to 3.23 {+-} 1.15 mmol/kg wet weight after correction. Average cell density of all meningiomas was 333 {+-} 119 cells/HPF, and average proliferation index was 2.93 {+-} 5.72%. A linear, positive correlation between cell density and Cho concentration was observed (r = 0.650, P = 0.001). After correction of Cho concentration, the correlation became more significant (r = 0.737, P < 0.001). However, no significant correlation between Cho concentration and proliferation index was found. There seemed to be a positive correlation trend after correction of Cho concentration but did not reach significant level. Absolute Cho concentration, especially Cho concentration corrected according to intra-voxel cystic/necrotic parts, reflects cell density of meningioma. (orig.)

  9. PyCorrFit-generic data evaluation for fluorescence correlation spectroscopy.

    Müller, Paul; Schwille, Petra; Weidemann, Thomas

    2014-09-01

    We present a graphical user interface (PyCorrFit) for the fitting of theoretical model functions to experimental data obtained by fluorescence correlation spectroscopy (FCS). The program supports many data file formats and features a set of tools specialized in FCS data evaluation. The Python source code is freely available for download from the PyCorrFit web page at http://pycorrfit.craban.de. We offer binaries for Ubuntu Linux, Mac OS X and Microsoft Windows. © The Author 2014. Published by Oxford University Press.

  10. Application of perturbed angular correlation spectroscopy in IgG immunoglobulins

    Silva, A.S.; Amaral, A.A.; Lapolli, A.L.; Carbonari, A.W.

    2009-01-01

    In the present work, the technique of perturbed angular correlation (PAC) spectroscopy was used to measure the electric hyperfine field at IgG immunoglobulins using 111 In → 111 Cd and 181 Hf → 181 Ta probe nuclei. The biological materials studied in this work were originating from the immunological response of different mice lineages infected by the Y strain of T. cruzi. The samples were measured at room temperature (295K) and at liquid nitrogen temperature (77K). The PAC results showed that, samples measured with 111 In obtained better results when they were compared with samples measured with 181 Hf. (author)

  11. Correlations between the orientation of magnetic recording media determined by Moessbauer spectroscopy and magnetic methods

    Pott, R.A.; Koch, W.; Leitner, L.

    1986-01-01

    The orientation of the easy magnetization axis of magnetic particles is a key parameter of the recording performance of magnetic recording media. Usually the orientation is measured by magnetic methods, but the applicability of the Moessbauer Spectroscopy has also been shown in the past. The authors show and discuss the correlations between the results obtained by magnetic and Moessbauer measurements for the example of several magnetic tapes. They demonstrate that by a combination of both methods one is even able to estimate the mean canting angles distribution width of the easy axis of magnetization. (Auth.)

  12. Application of perturbed angular correlation spectroscopy in IgG immunoglobulins

    Silva, A.S.; Amaral, A.A.; Lapolli, A.L.; Carbonari, A.W. [Instituto de Pesquisas Energeticas e Nucleares (IPEN-CNEN/SP), Sao Paulo, SP (Brazil)], e-mail: asilva@usp.br

    2009-07-01

    In the present work, the technique of perturbed angular correlation (PAC) spectroscopy was used to measure the electric hyperfine field at IgG immunoglobulins using {sup 111}In {yields}{sup 111}Cd and {sup 181}Hf {yields} {sup 181}Ta probe nuclei. The biological materials studied in this work were originating from the immunological response of different mice lineages infected by the Y strain of T. cruzi. The samples were measured at room temperature (295K) and at liquid nitrogen temperature (77K). The PAC results showed that, samples measured with {sup 111}In obtained better results when they were compared with samples measured with {sup 181}Hf. (author)

  13. CONTIN XPCS: Software for Inverse Transform Analysis of X-Ray Photon Correlation Spectroscopy Dynamics.

    Andrews, Ross N; Narayanan, Suresh; Zhang, Fan; Kuzmenko, Ivan; Ilavsky, Jan

    2018-02-01

    X-ray photon correlation spectroscopy (XPCS) and dynamic light scattering (DLS) both reveal dynamics using coherent scattering, but X-rays permit investigating of dynamics in a much more diverse array of materials. Heterogeneous dynamics occur in many such materials, and we showed how classic tools employed in analysis of heterogeneous DLS dynamics extend to XPCS, revealing additional information that conventional Kohlrausch exponential fitting obscures. This work presents the software implementation of inverse transform analysis of XPCS data called CONTIN XPCS, an extension of traditional CONTIN that accommodates dynamics encountered in equilibrium XPCS measurements.

  14. In vivo time-gated diffuse correlation spectroscopy at quasi-null source-detector separation.

    Pagliazzi, M; Sekar, S Konugolu Venkata; Di Sieno, L; Colombo, L; Durduran, T; Contini, D; Torricelli, A; Pifferi, A; Mora, A Dalla

    2018-06-01

    We demonstrate time domain diffuse correlation spectroscopy at quasi-null source-detector separation by using a fast time-gated single-photon avalanche diode without the need of time-tagging electronics. This approach allows for increased photon collection, simplified real-time instrumentation, and reduced probe dimensions. Depth discriminating, quasi-null distance measurement of blood flow in a human subject is presented. We envision the miniaturization and integration of matrices of optical sensors of increased spatial resolution and the enhancement of the contrast of local blood flow changes.

  15. Quantitative Studies of Antimicrobial Peptide Pore Formation in Large Unilamellar Vesicles by Fluorescence Correlation Spectroscopy (FCS)

    Kristensen, Kasper; Henriksen, Jonas Rosager; Andresen, Thomas Lars

    2013-01-01

    In spite of intensive research efforts over the past decades, the mechanisms by which membrane-active antimicrobial peptides interact with phospholipid membranes are not yet fully elucidated. New tools that can be used to characterize antimicrobial peptide-lipid membrane interactions are therefore...... to quantify leakage from large unilamellar vesicles is associated with a number of experimental pitfalls. Based on theoretical and experimental considerations, we discuss how to properly design experiments to avoid these pitfalls. Subsequently, we apply fluorescence correlation spectroscopy to quantify...

  16. Correlation analysis for forced vibration test of the Hualien large scale seismic test (LSST) program

    Sugawara, Y.; Sugiyama, T.; Kobayashi, T.; Yamaya, H.; Kitamura, E.

    1995-01-01

    The correlation analysis for a forced vibration test of a 1/4-scale containment SSI test model constructed in Hualien, Taiwan was carried out for the case of after backfilling. Prior to this correlation analysis, the structural properties were revised to adjust the calculated fundamental frequency in the fixed base condition to that derived from the test results. A correlation analysis was carried out using the Lattice Model which was able to estimate the soil-structure effects with embedment. The analysis results coincide well with test results and it is concluded that the mathematical soil-structure interaction model established by the correlation analysis is efficient in estimating the dynamic soil-structure interaction effect with embedment. This mathematical model will be applied as a basic model for simulation analysis of earthquake observation records. (author). 3 refs., 12 figs., 2 tabs

  17. Three-body correlations and conditional forces in suspensions of active hard disks

    Härtel, Andreas; Richard, David; Speck, Thomas

    2018-01-01

    Self-propelled Brownian particles show rich out-of-equilibrium physics, for instance, the motility-induced phase separation (MIPS). While decades of studying the structure of liquids have established a deep understanding of passive systems, not much is known about correlations in active suspensions. In this work we derive an approximate analytic theory for three-body correlations and forces in systems of active Brownian disks starting from the many-body Smoluchowski equation. We use our theory to predict the conditional forces that act on a tagged particle and their dependence on the propulsion speed of self-propelled disks. We identify preferred directions of these forces in relation to the direction of propulsion and the positions of the surrounding particles. We further relate our theory to the effective swimming speed of the active disks, which is relevant for the physics of MIPS. To test and validate our theory, we additionally run particle-resolved computer simulations, for which we explicitly calculate the three-body forces. In this context, we discuss the modeling of active Brownian swimmers with nearly hard interaction potentials. We find very good agreement between our simulations and numerical solutions of our theory, especially for the nonequilibrium pair-distribution function. For our analytical results, we carefully discuss their range of validity in the context of the different levels of approximation we applied. This discussion allows us to study the individual contribution of particles to three-body forces and to the emerging structure. Thus, our work sheds light on the collective behavior, provides the basis for further studies of correlations in active suspensions, and makes a step towards an emerging liquid state theory.

  18. Detrended partial cross-correlation analysis of two nonstationary time series influenced by common external forces

    Qian, Xi-Yuan; Liu, Ya-Min; Jiang, Zhi-Qiang; Podobnik, Boris; Zhou, Wei-Xing; Stanley, H. Eugene

    2015-06-01

    When common factors strongly influence two power-law cross-correlated time series recorded in complex natural or social systems, using detrended cross-correlation analysis (DCCA) without considering these common factors will bias the results. We use detrended partial cross-correlation analysis (DPXA) to uncover the intrinsic power-law cross correlations between two simultaneously recorded time series in the presence of nonstationarity after removing the effects of other time series acting as common forces. The DPXA method is a generalization of the detrended cross-correlation analysis that takes into account partial correlation analysis. We demonstrate the method by using bivariate fractional Brownian motions contaminated with a fractional Brownian motion. We find that the DPXA is able to recover the analytical cross Hurst indices, and thus the multiscale DPXA coefficients are a viable alternative to the conventional cross-correlation coefficient. We demonstrate the advantage of the DPXA coefficients over the DCCA coefficients by analyzing contaminated bivariate fractional Brownian motions. We calculate the DPXA coefficients and use them to extract the intrinsic cross correlation between crude oil and gold futures by taking into consideration the impact of the U.S. dollar index. We develop the multifractal DPXA (MF-DPXA) method in order to generalize the DPXA method and investigate multifractal time series. We analyze multifractal binomial measures masked with strong white noises and find that the MF-DPXA method quantifies the hidden multifractal nature while the multifractal DCCA method fails.

  19. Renormalization of correlations in a quasiperiodically forced two-level system: quadratic irrationals

    Mestel, B D; Osbaldestin, A H

    2004-01-01

    Generalizing from the case of golden mean frequency to a wider class of quadratic irrationals, we extend our renormalization analysis of the self-similarity of correlation functions in a quasiperiodically forced two-level system. We give a description of all piecewise-constant periodic orbits of an additive functional recurrence generalizing that present in the golden mean case. We establish a criterion for periodic orbits to be globally bounded, and also calculate the asymptotic height of the main peaks in the correlation function

  20. Combined IR-Raman vs vibrational sum-frequency heterospectral correlation spectroscopy

    Roy, Sandra; Beutier, Clémentine; Hore, Dennis K.

    2018-06-01

    Vibrational sum-frequency generation spectroscopy is a valuable probe of surface structure, particularly when the same molecules are present in one of the adjacent bulk solid or solution phases. As a result of the non-centrosymmetric requirement of SFG, the signal generated is a marker of the extent to which the molecules are ordered in an arrangement that breaks the up-down symmetry at the surface. In cases where the accompanying changes in the bulk are of interest in understanding and interpreting the surface structure, simultaneous analysis of the bulk IR absorption or bulk Raman scattering is helpful, and may be used in heterospectral surface-bulk two-dimensional correlation. We demonstrate that, in such cases, generating a new type of bulk spectrum that combines the IR and Raman amplitudes is a better candidate than the individual IR and Raman spectra for the purpose of correlation with the SFG signal.

  1. Probing the photoluminescence properties of gold nanoclusters by fluorescence lifetime correlation spectroscopy

    Yuan, C. T.; Lin, T. N.; Shen, J. L.; Lin, C. A.; Chang, W. H.; Cheng, H. W.; Tang, J.

    2013-01-01

    Gold nanoclusters (Au NCs) have attracted much attention for promising applications in biological imaging owing to their tiny sizes and biocompatibility. So far, most efforts have been focused on the strategies for fabricating high-quality Au NCs and then characterized by conventional ensemble measurement. Here, a fusion single-molecule technique combining fluorescence correlation spectroscopy and time-correlated single-photon counting can be successfully applied to probe the photoluminescence (PL) properties for sparse Au NCs. In this case, the triplet-state dynamics and diffusion process can be observed simultaneously and the relevant time constants can be derived. This work provides a complementary insight into the PL mechanism at the molecular levels for Au NCs in solution

  2. Temperature-Corrected Oxygen Detection Based on Multi-Mode Diode Laser Correlation Spectroscopy

    Xiutao Lou

    2013-01-01

    Full Text Available Temperature-corrected oxygen measurements were performed by using multi-mode diode laser correlation spectroscopy at temperatures ranging between 300 and 473 K. The experiments simulate in situ monitoring of oxygen in coal-combustion exhaust gases at the tail of the flue. A linear relationship with a correlation coefficient of −0.999 was found between the evaluated concentration and the gas temperature. Temperature effects were either auto-corrected by keeping the reference gas at the same conditions as the sample gas, or rectified by using a predetermined effective temperature-correction coefficient calibrated for a range of absorption wavelengths. Relative standard deviations of the temperature-corrected oxygen concentrations obtained by different schemes and at various temperatures were estimated, yielding a measurement precision of 0.6%.

  3. Correlation between laser-induced breakdown spectroscopy signal and moisture content

    Liu, Yuan [Townes Laser Institute, CREOL - The College of Optics and Photonics, University of Central Florida, 4000 Central Florida Boulevard, Orlando, FL 32816 (United States); Gigant, Lionel [Townes Laser Institute, CREOL - The College of Optics and Photonics, University of Central Florida, 4000 Central Florida Boulevard, Orlando, FL 32816 (United States); Universite Bordeaux 1, 351 cours de la Liberation 33405 Talence Cedex (France); Baudelet, Matthieu, E-mail: baudelet@creol.ucf.edu [Townes Laser Institute, CREOL - The College of Optics and Photonics, University of Central Florida, 4000 Central Florida Boulevard, Orlando, FL 32816 (United States); Richardson, Martin [Townes Laser Institute, CREOL - The College of Optics and Photonics, University of Central Florida, 4000 Central Florida Boulevard, Orlando, FL 32816 (United States)

    2012-07-15

    The possibility of using Laser-Induced Breakdown Spectroscopy (LIBS) for measuring the moisture content of fresh food samples is studied. The normalized line emission of oxygen is highly correlated with the moisture content of the sample, cheese in our case, and can be used as a moisture marker in situations where oxygen interference from the matrix is not a critical issue. The linear correlation between the oxygen signal and the moisture content in the sample shows great potential for using LIBS as an alternative spectroscopic method for moisture monitoring. - Highlights: Black-Right-Pointing-Pointer Quantitative moisture measurement by LIBS. Black-Right-Pointing-Pointer Use of matrix effects and normalization for physical information on the sample. Black-Right-Pointing-Pointer Use of signal from oxygen and CN radical in air background for moisture measurement.

  4. A method of Moessbauer Fourier spectroscopy for determination of the biopolimer coordinate correlation functions

    Basovets, S.K.; Krupyanskij, Yu.F.; Kurinov, I.V.; Suzdalev, I.P.; Goldanskij, V.I.; Uporov, I.V.; Shaitan, K.V.; Rubin, A.B.

    1988-01-01

    A method of Moessbauer Fourier spectroscopy is developed to determine the correlation function of coordinates of a macromolecular system. The method does not require the use of an a priori dynamic model. The application of the method to the analysis of RSMR data for human serum albumin has demonstrated considerable changes in the dynamic behavior of the protein globule when the temperature is changed from 270 to 310 K. The main conclusions of the present work is the simultaneous observation of low-frequency (τ≥10 -9 sec) and high-frequency (τ -9 sec) large-scaled motions, that is the two-humped distribution of correlation times of protein motions. (orig.)

  5. Correlation between laser-induced breakdown spectroscopy signal and moisture content

    Liu, Yuan; Gigant, Lionel; Baudelet, Matthieu; Richardson, Martin

    2012-01-01

    The possibility of using Laser-Induced Breakdown Spectroscopy (LIBS) for measuring the moisture content of fresh food samples is studied. The normalized line emission of oxygen is highly correlated with the moisture content of the sample, cheese in our case, and can be used as a moisture marker in situations where oxygen interference from the matrix is not a critical issue. The linear correlation between the oxygen signal and the moisture content in the sample shows great potential for using LIBS as an alternative spectroscopic method for moisture monitoring. - Highlights: ► Quantitative moisture measurement by LIBS. ► Use of matrix effects and normalization for physical information on the sample. ► Use of signal from oxygen and CN radical in air background for moisture measurement.

  6. Correlations Between Magnetic Flux and Levitation Force of HTS Bulk Above a Permanent Magnet Guideway

    Huang, Huan; Zheng, Jun; Zheng, Botian; Qian, Nan; Li, Haitao; Li, Jipeng; Deng, Zigang

    2017-10-01

    In order to clarify the correlations between magnetic flux and levitation force of the high-temperature superconducting (HTS) bulk, we measured the magnetic flux density on bottom and top surfaces of a bulk superconductor while vertically moving above a permanent magnet guideway (PMG). The levitation force of the bulk superconductor was measured simultaneously. In this study, the HTS bulk was moved down and up for three times between field-cooling position and working position above the PMG, followed by a relaxation measurement of 300 s at the minimum height position. During the whole processes, the magnetic flux density and levitation force of the bulk superconductor were recorded and collected by a multipoint magnetic field measurement platform and a self-developed maglev measurement system, respectively. The magnetic flux density on the bottom surface reflected the induced field in the superconductor bulk, while on the top, it reveals the penetrated magnetic flux. The results show that the magnetic flux density and levitation force of the bulk superconductor are in direct correlation from the viewpoint of inner supercurrent. In general, this work is instructive for understanding the connection of the magnetic flux density, the inner current density and the levitation behavior of HTS bulk employed in a maglev system. Meanwhile, this magnetic flux density measurement method has enriched present experimental evaluation methods of maglev system.

  7. NRC test results and operations experience provide insights for a new gate valve stem force correlation

    Watkins, John C.; Steele, Robert Jr.; DeWall, Kevin G.; Weidenhamer, G.H.; Rothberg, O.O.

    1994-01-01

    This paper presents the results of testing sponsored by the NRC to assess valve and motor operator performance under varying pressure and fluid conditions. This effort included an examination of the methods used by the industry to predict the required stem force of a valve, and research to provide guidelines for the extrapolation of in situ test results to design basis conditions.Years ago, when most of these valves were originally installed, the industry used a set of equations to determine analytically that the valves' motor-operators were large enough and the control switches were set high enough to close the valves at their design basis conditions. Our research has identified several inconsistencies with the industry's existing gate valve stem force equation and has challenged the overly simplistic assumptions inherent in its use. This paper discusses the development of the INEL correlation, which serves as the basis for a method to bound the stem force necessary to close flexwedge gate valves whose operational characteristics have been shown to be predictable. As utilities undertake to provide assurance of their valves' operability, this ability to predict analytically the required stem force is especially important for valves that cannot be tested at design basis conditions. For such valves, the results of tests conducted at less severe conditions can be used with the INEL correlation to make the necessary prediction. ((orig.))

  8. Condom Use Behaviors and Correlates of Use in the Botswana Defence Force

    Tran, Bonnie Robin; Thomas, Anne Goldzier; Ditsela, Mooketsi; Vaida, Florin; Phetogo, Robert; Kelapile, David; Chambers, Christina; Haubrich, Richard; Shaffer, Richard

    2013-01-01

    Preventing HIV infection is a priority for militaries. HIV prevention research is needed to monitor existing programs, identify areas for modification, and develop new interventions. Correct and consistent condom use is highly effective against HIV. However, use among soldiers is lower than ideal. This study describes condom use behaviors and examines correlates of use in the Botswana Defence Force (BDF). Analyses were based on 211 male personnel, aged 18–30, who completed a cross-sectional s...

  9. Two-dimensional MR spectroscopy of minimal hepatic encephalopathy and neuropsychological correlates in vivo.

    Singhal, Aparna; Nagarajan, Rajakumar; Hinkin, Charles H; Kumar, Rajesh; Sayre, James; Elderkin-Thompson, Virginia; Huda, Amir; Gupta, Rakesh K; Han, Steven-Huy; Thomas, M Albert

    2010-07-01

    To evaluate regional cerebral metabolic and structural changes in patients with minimal hepatic encephalopathy (MHE) using two-dimensional (2D) MR spectroscopy (MRS) and T( (1) )-weighted MRI, to correlate the observed MR changes with neuropsychological (NP) test scores, and to compare the diagnostic accuracy of MRI, 2D MRS, and NP tests in discriminating between patients and healthy subjects. Thirty-three MHE patients and 30 healthy controls were investigated. The 2D localized correlated spectroscopy (L-COSY) was performed in the frontal and occipital brain on a 1.5 Tesla (T) MR scanner. The NP test battery included 15 tests, grouped into 6 cognitive domains. Globus pallidus signal intensities were calculated from T(1)-weighted images. The 2D MRS showed significant differences in ratios of the following metabolite(s) peaks with respect to creatine (Cr): decreased myo-inositol (mI), choline (Ch), mICh, and increased (glutamate plus glutamine) (Glx) in patients compared with healthy subjects in both occipital and frontal lobes. Frontal lobe taurine also showed a decline in patients. The NP test results revealed declines in cognitive speed, motor function, executive function, and global cognitive status. Significant correlations were found between the altered metabolites and NP tests. Alteration in the mICh/Cr ratio was noted as a powerful discriminant between healthy subjects and the patients. The study demonstrates that relative metabolite levels determined by 2D MRS, in particular mICh/Cr, provide the best diagnostic prediction for MHE. The results suggest that depletions of myo-inositol, choline and taurine with respect to creatine correlate with measures of neuropsychological impairment. (c) 2010 Wiley-Liss, Inc.

  10. Tip-surface interactions at redox responsive poly(ferrocenylsilane) (PFS) interface by AFM-based force spectroscopy

    Chung Hongjing; Song Jing; Vancso, G. Julius

    2009-01-01

    Poly(ferrocenylsilanes) (PFS) belong to the class of redox responsive organometallic polymers. Atomic force microscopy (AFM)-based single molecule force spectroscopy (SMFS) was used earlier to study single chain PFS response and redox energy driven single chain PFS molecular motors. Here we present further AFM investigations of force interactions between tip and a grafted PFS surface under potential control in electrochemical redox cycles. Typical tip-Au interaction is considered as reference in the force measurements. First the electrostatic component in the diffused double layer (DL) in NaClO 4 electrolyte environment was considered for a 'grafted to' PFS, which dominated the interplay between the tip and sample surface. The DL forces can also hinder the physisorption of PFS chain onto the tip when the voltage was applied at -0.1 V. On the other hand, if the tip contacted the PFS surface prior to the electrochemical process, physisorption of PFS chains governed the overall interaction regardless of subsequently applied surface potential. In addition, prolonged contact time, t c , may also contribute to the stability of tip-PFS bridging and detection of electrostatic forces between the tip-PFS interface. The results showed that tip-substrate interaction forces without PFS grafts have negligibly small force contributions under similar, electrochemically controlled, conditions used in single PFS chain based molecular motors.

  11. Accelerating two-dimensional nuclear magnetic resonance correlation spectroscopy via selective coherence transfer

    Ye, Qimiao; Chen, Lin; Qiu, Wenqi; Lin, Liangjie; Sun, Huijun; Cai, Shuhui; Wei, Zhiliang; Chen, Zhong

    2017-01-01

    Nuclear magnetic resonance (NMR) spectroscopy serves as an important tool for both qualitative and quantitative analyses of various systems in chemistry, biology, and medicine. However, applications of one-dimensional 1H NMR are often restrained by the presence of severe overlap among different resonances. The advent of two-dimensional (2D) 1H NMR constitutes a promising alternative by extending the crowded resonances into a plane and thereby alleviating the spectral congestions. However, the enhanced ability in discriminating resonances is achieved at the cost of extended experimental duration due to necessity of various scans with progressive delays to construct the indirect dimension. Therefore, in this study, we propose a selective coherence transfer (SECOT) method to accelerate acquisitions of 2D correlation spectroscopy by converting chemical shifts into spatial positions within the effective sample length and then performing an echo planar spectroscopic imaging module to record the spatial and spectral information, which generates 2D correlation spectrum after 2D Fourier transformation. The feasibility and effectiveness of SECOT have been verified by a set of experiments under both homogeneous and inhomogeneous magnetic fields. Moreover, evaluations of SECOT for quantitative analyses are carried out on samples with a series of different concentrations. Based on these experimental results, the SECOT may open important perspectives for fast, accurate, and stable investigations of various chemical systems both qualitatively and quantitatively.

  12. Intracellular localization and dynamics of Hypericin loaded PLLA nanocarriers by image correlation spectroscopy.

    Penjweini, Rozhin; Deville, Sarah; D'Olieslaeger, Lien; Berden, Mandy; Ameloot, Marcel; Ethirajan, Anitha

    2015-11-28

    The study of cell-nanoparticle interactions is an important aspect for understanding drug delivery using nanocarriers. In this regard, advances in fluorescence based microscopy are useful for the investigation of temporal and spatial behavior of nanoparticles (NPs) within the intracellular environment. In this work, we focus on the delivery of the naturally-occurring hydrophobic photosensitizer Hypericin in human lung carcinoma A549 cells by using biodegradable poly L-lactic acid NPs. For the first time, Hypericin containing NPs are prepared by combining the miniemulsion technique with the solvent evaporation method. This approach yields an efficient loading of the NPs with Hypericin and allows for additional cargo molecules. To monitor the release of Hypercin from the NPs, an additional fluorescent lipophilic dye Coumarin-6 is incorporated in the NPs. Temporal and spatiotemporal image correlation spectroscopy is used to determine the fate of the NPs carrying the potential cargo. Both directed and non-directed motions are detected. By using image cross-correlation spectroscopy and specific fluorescent labeling of endosomes, lysosomes and mitochondria, the dynamics of the cargo loaded NPs in association with the organelles is studied. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Sensorimotor and neuropsychological correlates of force perturbations that induce stepping in older adults.

    Sturnieks, Daina L; Menant, Jasmine; Vanrenterghem, Jos; Delbaere, Kim; Fitzpatrick, Richard C; Lord, Stephen R

    2012-07-01

    Inappropriate stepping in response to unexpected balance perturbations is more prevalent in older people and in those at risk of falling. This study examined responses to force-controlled waist pulls in young and older people, and sought to identify physiological and cognitive correlates of the force threshold for stepping. 242 older (79.7±4.2 years) and 15 young (29.5±5.3 years) adults underwent waist pull perturbations and assessments of physiological and neuropsychological functioning, general health and falls efficacy. Perturbation force that induced stepping, stepping strategy and number of steps were measured. The older group withstood less forceful perturbations with a feet-in-place strategy, compared to young. Likewise, older adults with high falls risk withstood less force than those with low risk. After controlling for body weight and gender, sway and lower limb strength were independent predictors of anterior stepping thresholds, reaction time was an independent predictor of posterior thresholds, and executive functioning and lower limb strength were independent predictors of the lateral thresholds. These results suggest that balance, strength and agility training, in addition to cognitive exercises may enhance the ability to withstand unexpected balance perturbations and reduce the risk of falls in older people. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. A novel aptasensor based on single-molecule force spectroscopy for highly sensitive detection of mercury ions.

    Li, Qing; Michaelis, Monika; Wei, Gang; Colombi Ciacchi, Lucio

    2015-08-07

    We have developed a novel aptasensor based on single-molecule force spectroscopy (SMFS) capable of detecting mercury ions (Hg(2+)) with sub-nM sensitivity. The single-strand (ss) DNA aptamer used in this work is rich in thymine (T) and readily forms T-Hg(2+)-T complexes in the presence of Hg(2+). The aptamer was conjugated to an atomic force microscope (AFM) probe, and the adhesion force between the probe and a flat graphite surface was measured by single-molecule force spectroscopy (SMFS). The presence of Hg(2+) ions above a concentration threshold corresponding to the affinity constant of the ions for the aptamer (about 5 × 10(9) M(-1)) could be easily detected by a change of the measured adhesion force. With our chosen aptamer, we could reach an Hg(2+) detection limit of 100 pM, which is well below the maximum allowable level of Hg(2+) in drinking water. In addition, this aptasensor presents a very high selectivity for Hg(2+) over other metal cations, such as K(+), Ca(2+), Zn(2+), Fe(2+), and Cd(2+). Furthermore, the effects of the ionic strength and loading rate on the Hg(2+) detection were evaluated. Its simplicity, reproducibility, high selectivity and sensitivity make our SMFS-based aptasensor advantageous with respect to other current Hg(2+) sensing methods. It is expected that our strategy can be exploited for monitoring the pollution of water environments and the safety of potentially contaminated food.

  15. Insights into the Interactions of Amino Acids and Peptides with Inorganic Materials Using Single-Molecule Force Spectroscopy.

    Das, Priyadip; Duanias-Assaf, Tal; Reches, Meital

    2017-03-06

    The interactions between proteins or peptides and inorganic materials lead to several interesting processes. For example, combining proteins with minerals leads to the formation of composite materials with unique properties. In addition, the undesirable process of biofouling is initiated by the adsorption of biomolecules, mainly proteins, on surfaces. This organic layer is an adhesion layer for bacteria and allows them to interact with the surface. Understanding the fundamental forces that govern the interactions at the organic-inorganic interface is therefore important for many areas of research and could lead to the design of new materials for optical, mechanical and biomedical applications. This paper demonstrates a single-molecule force spectroscopy technique that utilizes an AFM to measure the adhesion force between either peptides or amino acids and well-defined inorganic surfaces. This technique involves a protocol for attaching the biomolecule to the AFM tip through a covalent flexible linker and single-molecule force spectroscopy measurements by atomic force microscope. In addition, an analysis of these measurements is included.

  16. Gamma Spectroscopy

    Niemantsverdriet, J.W.; Butz, Tilman; Ertl, G.; Knözinger, H.; Schüth, F.

    2008-01-01

    No abstract. The sections in this article are 1 Introduction 2 Mössbauer Spectroscopy 3 Time-Differential Perturbed Angular Correlations (TDPAC) 4 Conclusions and Outlook Keywords: Mössbauer spectroscopy; gamma spectroscopy; perturbed angular correlation; TDPAC

  17. The Correlation Between Dislocations and Vacancy Defects Using Positron Annihilation Spectroscopy

    Pang, Jinbiao; Li, Hui; Zhou, Kai; Wang, Zhu

    2012-07-01

    An analysis program for positron annihilation lifetime spectra is only applicable to isolated defects, but is of no use in the presence of defective correlations. Such limitations have long caused problems for positron researchers in their studies of complicated defective systems. In order to solve this problem, we aim to take a semiconductor material, for example, to achieve a credible average lifetime of single crystal silicon under plastic deformation at different temperatures using positron life time spectroscopy. By establishing reasonable positron trapping models with defective correlations and sorting out four lifetime components with multiple parameters, as well as their respective intensities, information is obtained on the positron trapping centers, such as the positron trapping rates of defects, the density of the dislocation lines and correlation between the dislocation lines, and the vacancy defects, by fitting with the average lifetime with the aid of Matlab software. These results give strong grounds for the existence of dislocation-vacancy correlation in plastically deformed silicon, and lay a theoretical foundation for the analysis of positron lifetime spectra when the positron trapping model involves dislocation-related defects.

  18. Fourier Transform Infrared (FTIR) Spectroscopy, Ultraviolet Resonance Raman (UVRR) Spectroscopy, and Atomic Force Microscopy (AFM) for Study of the Kinetics of Formation and Structural Characterization of Tau Fibrils.

    Ramachandran, Gayathri

    2017-01-01

    Kinetic studies of tau fibril formation in vitro most commonly employ spectroscopic probes such as thioflavinT fluorescence and laser light scattering or negative stain transmission electron microscopy. Here, I describe the use of Fourier transform infrared (FTIR) spectroscopy, ultraviolet resonance Raman (UVRR) spectroscopy, and atomic force microscopy (AFM) as complementary probes for studies of tau aggregation. The sensitivity of vibrational spectroscopic techniques (FTIR and UVRR) to secondary structure content allows for measurement of conformational changes that occur when the intrinsically disordered protein tau transforms into cross-β-core containing fibrils. AFM imaging serves as a gentle probe of structures populated over the time course of tau fibrillization. Together, these assays help further elucidate the structural and mechanistic complexity inherent in tau fibril formation.

  19. Ultra-short laser pulse ablation using shear-force feedback: Femtosecond laser induced breakdown spectroscopy feasibility study

    Samek, Ota; Kurowski, Andre; Kittel, Silke; Kukhlevsky, Sergei; Hergenroeder, Roland

    2005-01-01

    This work reports on a feasibility study of proximity ablation using femtosecond pulses. Ultra-short pulses were launched to a bare tapered optical fiber and delivered to the sample. The tip-sample distance was controlled by means of shear-force feedback. Consequently, ablation craters with submicrometer dimensions were obtained. Potential analytical applications for Laser Induced Breakdown Spectroscopy (LIBS) technique, such as e.g. inclusions in steel or bio cells, are suggested

  20. Dissipative and electrostatic force spectroscopy of indium arsenide quantum dots by non-contact atomic force microscopy

    Stomp, Romain-Pierre

    This thesis is devoted to the studies of self-assembled InAs quantum dots (QD) by low-temperature Atomic Force Microscopy (AFM) in frequency modulation mode. Several spectroscopic methods are developed to investigate single electron charging from a two-dimensional electron gas (2DEG) to an individual InAs QD. Furthermore, a new technique to measure the absolute tip-sample capacitance is also demonstrated. The main observables are the electrostatic force between the metal-coated AFM tip and sample as well as the sample-induced energy dissipation, and therefore no tunneling current has to be collected at the AFM tip. Measurements were performed by recording simultaneously the shift in the resonant frequency and the Q-factor degradation of the oscillating cantilever either as a function of tip-sample voltage or distance. The signature of single electron charging was detected as an abrupt change in the frequency shift as well as corresponding peaks in the dissipation. The main experimental features in the force agree well with the semi-classical theory of Coulomb blockade by considering the free energy of the system. The observed dissipation peaks can be understood as a back-action effect on the oscillating cantilever beam due to the fluctuation in time of electrons tunneling back and forth between the 2DEG and the QD. It was also possible to extract the absolute value of the tip-sample capacitance, as a consequence of the spectroscopic analysis of the electrostic force as a function of tip-sample distance for different values of the applied voltage. At the same time, the contact potential difference and the residual non-capacitive force could also be determined as a function of tip-sample distance.

  1. Chemical profiling and adulteration screening of Aquilariae Lignum Resinatum by Fourier transform infrared (FT-IR) spectroscopy and two-dimensional correlation infrared (2D-IR) spectroscopy

    Qu, Lei; Chen, Jian-bo; Zhang, Gui-Jun; Sun, Su-qin; Zheng, Jing

    2017-03-01

    As a kind of expensive perfume and valuable herb, Aquilariae Lignum Resinatum (ALR) is often adulterated for economic motivations. In this research, Fourier transform infrared (FT-IR) spectroscopy is employed to establish a simple and quick method for the adulteration screening of ALR. First, the principal chemical constituents of ALR are characterized by FT-IR spectroscopy at room temperature and two-dimensional correlation infrared (2D-IR) spectroscopy with thermal perturbation. Besides the common cellulose and lignin compounds, a certain amount of resin is the characteristic constituent of ALR. Synchronous and asynchronous 2D-IR spectra indicate that the resin (an unstable secondary metabolite) is more sensitive than cellulose and lignin (stable structural constituents) to the thermal perturbation. Using a certified ALR sample as the reference, the infrared spectral correlation threshold is determined by 30 authentic samples and 6 adulterated samples. The spectral correlation coefficient of an authentic ALR sample to the standard reference should be not less than 0.9886 (p = 0.01). Three commercial adulterated ALR samples are identified by the correlation threshold. Further interpretation of the infrared spectra of the adulterated samples indicates the common adulterating methods - counterfeiting with other kind of wood, adding ingredient such as sand to increase the weight, and adding the cheap resin such as rosin to increase the content of resin compounds. Results of this research prove that FT-IR spectroscopy can be used as a simple and accurate quality control method of ALR.

  2. Probing the nanoscale interaction forces and elastic properties of organic and inorganic materials using force-distance (F-D) spectroscopy

    Vincent, Abhilash

    Due to their therapeutic applications such as radical scavenging, MRI contrast imaging, Photoluminescence imaging, drug delivery, etc., nanoparticles (NPs) have a significant importance in bio-nanotechnology. The reason that prevents the utilizing NPs for drug delivery in medical field is mostly due to their biocompatibility issues (incompatibility can lead to toxicity and cell death). Changes in the surface conditions of NPs often lead to NP cytotoxicity. Investigating the role of NP surface properties (surface charges and surface chemistry) on their interactions with biomolecules (Cells, protein and DNA) could enhance the current understanding of NP cytotoxicity. Hence, it is highly beneficial to the nanotechnology community to bring more attention towards the enhancement of surface properties of NPs to make them more biocompatible and less toxic to biological systems. Surface functionalization of NPs using specific ligand biomolecules have shown to enhance the protein adsorption and cellular uptake through more favorable interaction pathways. Cerium oxide NPs (CNPs also known as nanoceria) are potential antioxidants in cell culture models and understanding the nature of interaction between cerium oxide NPs and biological proteins and cells are important due to their therapeutic application (especially in site specific drug delivery systems). The surface charges and surface chemistry of CNPs play a major role in protein adsorption and cellular uptake. Hence, by tuning the surface charges and by selecting proper functional molecules on the surface, CNPs exhibiting strong adhesion to biological materials can be prepared. By probing the nanoscale interaction forces acting between CNPs and protein molecules using Atomic Force Microscopy (AFM) based force-distance (F-D) spectroscopy, the mechanism of CNP-protein adsorption and CNP cellular uptake can be understood more quantitatively. The work presented in this dissertation is based on the application of AFM in

  3. Childhood temporal lobe epilepsy: correlation between electroencephalography and magnetic resonance spectroscopy: a case-control study.

    Azab, Seham Fa; Sherief, Laila M; Saleh, Safaa H; Elshafeiy, Mona M; Siam, Ahmed G; Elsaeed, Wafaa F; Arafa, Mohamed A; Bendary, Eman A; Sherbiny, Hanan S; Elbehedy, Rabab M; Aziz, Khalid A

    2015-04-18

    The diagnosis of epilepsy should be made as early as possible to give a child the best chance for treatment success and also to decrease complications such as learning difficulties and social and behavioral problems. In this study, we aimed to assess the ability of magnetic resonance spectroscopy (MRS) in detecting the lateralization side in patients with Temporal lobe epilepsy (TLE) in correlation with EEG and MRI findings. This was a case-control study including 40 patients diagnosed (clinically and by EEG) as having temporal lobe epilepsy aged 8 to 14 years (mean, 10.4 years) and 20 healthy children with comparable age and gender as the control group. All patients were subjected to clinical examination, interictal electroencephalography and magnetic resonance imaging (MRI). Proton magnetic resonance spectroscopic examination (MRS) was performed to the patients and the controls. According to the findings of electroencephalography, our patients were classified to three groups: Group 1 included 20 patients with unitemporal (lateralized) epileptic focus, group 2 included 12 patients with bitemporal (non-lateralized) epileptic focus and group 3 included 8 patients with normal electroencephalography. Magnetic resonance spectroscopy could lateralize the epileptic focus in 19 patients in group 1, nine patients in group2 and five patients in group 3 with overall lateralization of (82.5%), while electroencephalography was able to lateralize the focus in (50%) of patients and magnetic resonance imaging detected lateralization of mesial temporal sclerosis in (57.5%) of patients. Magnetic resonance spectroscopy is a promising tool in evaluating patients with epilepsy and offers increased sensitivity to detect temporal pathology that is not obvious on structural MRI imaging.

  4. A nu-space for image correlation spectroscopy: characterization and application to measure protein transport in live cells

    Potvin-Trottier, Laurent; Chen, Lingfeng; Horwitz, Alan Rick; Wiseman, Paul W.

    2013-08-01

    We introduce a new generalized theoretical framework for image correlation spectroscopy (ICS). Using this framework, we extend the ICS method in time-frequency (ν, nu) space to map molecular flow of fluorescently tagged proteins in individual living cells. Even in the presence of a dominant immobile population of fluorescent molecules, nu-space ICS (nICS) provides an unbiased velocity measurement, as well as the diffusion coefficient of the flow, without requiring filtering. We also develop and characterize a tunable frequency-filter for spatio-temporal ICS (STICS) that allows quantification of the density, the diffusion coefficient and the velocity of biased diffusion. We show that the techniques are accurate over a wide range of parameter space in computer simulation. We then characterize the retrograde flow of adhesion proteins (α6- and αLβ2-GFP integrins and mCherry-paxillin) in CHO.B2 cells plated on laminin and intercellular adhesion molecule 1 (ICAM-1) ligands respectively. STICS with a tunable frequency filter, in conjunction with nICS, measures two new transport parameters, the density and transport bias coefficient (a measure of the diffusive character of a flow/biased diffusion), showing that molecular flow in this cell system has a significant diffusive component. Our results suggest that the integrin-ligand interaction, along with the internal myosin-motor generated force, varies for different integrin-ligand pairs, consistent with previous results.

  5. Fluorescence Spectroscopy Applied to Monitoring Biodiesel Degradation: Correlation with Acid Value and UV Absorption Analyses.

    Vasconcelos, Maydla Dos Santos; Passos, Wilson Espíndola; Lescanos, Caroline Honaiser; Pires de Oliveira, Ivan; Trindade, Magno Aparecido Gonçalves; Caires, Anderson Rodrigues Lima; Muzzi, Rozanna Marques

    2018-01-01

    The techniques used to monitor the quality of the biodiesel are intensely discussed in the literature, partly because of the different oil sources and their intrinsic physicochemical characteristics. This study aimed to monitor the thermal degradation of the fatty acid methyl esters of Sesamum indicum L. and Raphanus sativus L. biodiesels (SILB and RSLB, resp.). The results showed that both biodiesels present a high content of unsaturated fatty acids, ∼84% (SILB) and ∼90% (RSLB). The SILB had a high content of polyunsaturated linoleic fatty acid (18  :  2), about 49%, and the oleic monounsaturated (18  :  1), ∼34%. On the other hand, RSLB presented a considerable content of linolenic fatty acid (18  :  3), ∼11%. The biodiesel samples were thermal degraded at 110°C for 48 hours, and acid value, UV absorption, and fluorescence spectroscopy analysis were carried out. The results revealed that both absorption and fluorescence presented a correlation with acid value as a function of degradation time by monitoring absorptions at 232 and 270 nm as well as the emission at 424 nm. Although the obtained correlation is not completely linear, a direct correlation was observed in both cases, revealing that both properties can be potentially used for monitoring the biodiesel degradation.

  6. Insight into resolution enhancement in generalized two-dimensional correlation spectroscopy.

    Ma, Lu; Sikirzhytski, Vitali; Hong, Zhenmin; Lednev, Igor K; Asher, Sanford A

    2013-03-01

    Generalized two-dimensional correlation spectroscopy (2D-COS) can be used to enhance spectral resolution in order to help differentiate highly overlapped spectral bands. Despite the numerous extensive 2D-COS investigations, the origin of the 2D spectral resolution enhancement mechanism(s) is not completely understood. In the work here, we studied the 2D-COS of simulated spectra in order to develop new insights into the dependence of 2D-COS spectral features on the overlapping band separations, their intensities and bandwidths, and their band intensity change rates. We found that the features in the 2D-COS maps that are derived from overlapping bands were determined by the spectral normalized half-intensities and the total intensity changes of the correlated bands. We identified the conditions required to resolve overlapping bands. In particular, 2D-COS peak resolution requires that the normalized half-intensities of a correlating band have amplitudes between the maxima and minima of the normalized half-intensities of the overlapping bands.

  7. Probing GFP-actin diffusion in living cells using fluorescence correlation spectroscopy

    Engelke, Hanna; Heinrich, Doris; Rädler, Joachim O.

    2010-01-01

    The cytoskeleton of eukaryotic cells is continuously remodeled by polymerization and depolymerization of actin. Consequently, the relative content of polymerized filamentous actin (F-actin) and monomeric globular actin (G-actin) is subject to temporal and spatial fluctuations. Since fluorescence correlation spectroscopy (FCS) can measure the diffusion of fluorescently labeled actin it seems likely that FCS allows us to determine the dynamics and hence indirectly the structural properties of the cytoskeleton components with high spatial resolution. To this end we investigate the FCS signal of GFP-actin in living Dictyostelium discoideum cells and explore the inherent spatial and temporal signatures of the actin cytoskeleton. Using the free green fluorescent protein (GFP) as a reference, we find that actin diffusion inside cells is dominated by G-actin and slower than diffusion in diluted cell extract. The FCS signal in the dense cortical F-actin network near the cell membrane is probed using the cytoskeleton protein LIM and is found to be slower than cytosolic G-actin diffusion. Furthermore, we show that polymerization of the cytoskeleton induced by Jasplakinolide leads to a substantial decrease of G-actin diffusion. Pronounced fluctuations in the distribution of the FCS correlation curves can be induced by latrunculin, which is known to induce actin waves. Our work suggests that the FCS signal of GFP-actin in combination with scanning or spatial correlation techniques yield valuable information about the local dynamics and concomitant cytoskeletal properties

  8. Anterior insula GABA levels correlate with emotional aspects of empathy: a proton magnetic resonance spectroscopy study.

    Qianfeng Wang

    Full Text Available Empathy is a multidimensional construct referring to the capacity to understand and share the emotional and affective states of another person. Cerebral γ-aminobutyric acid (GABA-ergic levels are associated with a variety of neurological and psychiatric disorders. However, the role of the GABA system in different dimensions of empathy has not been investigated.Thirty-two right-handed healthy volunteers took part in this study. We used proton magnetic resonance spectroscopy to determine GABA concentrations in the anterior insula (AI and the anterior cingulate cortex (ACC and to examine the relationship between the GABA concentrations and the subcomponents of empathy evaluated by the Interpersonal Reactivity Index (IRI.Pearson correlation analyses (two-tailed showed that AI GABA was significantly associated with the empathy concern score (r = 0.584, p<0.05 and the personal distress score (r = 0.538, p<0.05 but not significantly associated with other empathy subscales. No significant correlation was found between ACC GABA and empathy subscores.Left AI GABA was positively correlated with the emotional aspects of empathy. These preliminary findings call into question whether AI GABA alterations might predict empathy dysfunction in major psychiatric disorders such as autism and schizophrenia, which have been described as deficits in emotional empathic abilities.

  9. Fluorescence Spectroscopy Applied to Monitoring Biodiesel Degradation: Correlation with Acid Value and UV Absorption Analyses

    Maydla dos Santos Vasconcelos

    2018-01-01

    Full Text Available The techniques used to monitor the quality of the biodiesel are intensely discussed in the literature, partly because of the different oil sources and their intrinsic physicochemical characteristics. This study aimed to monitor the thermal degradation of the fatty acid methyl esters of Sesamum indicum L. and Raphanus sativus L. biodiesels (SILB and RSLB, resp.. The results showed that both biodiesels present a high content of unsaturated fatty acids, ∼84% (SILB and ∼90% (RSLB. The SILB had a high content of polyunsaturated linoleic fatty acid (18  :  2, about 49%, and the oleic monounsaturated (18  :  1, ∼34%. On the other hand, RSLB presented a considerable content of linolenic fatty acid (18  :  3, ∼11%. The biodiesel samples were thermal degraded at 110°C for 48 hours, and acid value, UV absorption, and fluorescence spectroscopy analysis were carried out. The results revealed that both absorption and fluorescence presented a correlation with acid value as a function of degradation time by monitoring absorptions at 232 and 270 nm as well as the emission at 424 nm. Although the obtained correlation is not completely linear, a direct correlation was observed in both cases, revealing that both properties can be potentially used for monitoring the biodiesel degradation.

  10. Spectroscopy

    Berg, Rolf W.

    This introductory booklet covers the basics of molecular spectroscopy, infrared and Raman methods, instrumental considerations, symmetry analysis of molecules, group theory and selection rules, as well as assignments of fundamental vibrational modes in molecules.......This introductory booklet covers the basics of molecular spectroscopy, infrared and Raman methods, instrumental considerations, symmetry analysis of molecules, group theory and selection rules, as well as assignments of fundamental vibrational modes in molecules....

  11. Forced convection heat transfer correlation for finned plates in a duct

    Chae, Myeong-Seon; Moon, Je-Young; Chung, Bum-Jin

    2014-01-01

    Forced convection heat transfer experiments were conducted for plate-fin in a duct using various fin spacing, fin height, duct width, Reynolds number for Prandtl numbers 2,014. Based upon analogy concept, mass transfer rate were measured instead of heat transfer rates. The heat transfer rates were enhanced with the increase of fin height and decrease of fin spacing as they increase the heat transfer area. Meanwhile, heat transfer rates were impaired with the increase of the duct width as the bypass flows increased to tip clearance region. Forced convection heat transfer correlations were developed for laminar and turbulent flow conditions and for narrow and wide ducts. The work draws attention to the tip clearance on the heat transfer of the finned plate in a duct. (author)

  12. Easy monitoring of velocity fields in microfluidic devices using spatiotemporal image correlation spectroscopy.

    Travagliati, Marco; Girardo, Salvatore; Pisignano, Dario; Beltram, Fabio; Cecchini, Marco

    2013-09-03

    Spatiotemporal image correlation spectroscopy (STICS) is a simple and powerful technique, well established as a tool to probe protein dynamics in cells. Recently, its potential as a tool to map velocity fields in lab-on-a-chip systems was discussed. However, the lack of studies on its performance has prevented its use for microfluidics applications. Here, we systematically and quantitatively explore STICS microvelocimetry in microfluidic devices. We exploit a simple experimental setup, based on a standard bright-field inverted microscope (no fluorescence required) and a high-fps camera, and apply STICS to map liquid flow in polydimethylsiloxane (PDMS) microchannels. Our data demonstrates optimal 2D velocimetry up to 10 mm/s flow and spatial resolution down to 5 μm.

  13. Comparison of nanoparticle diffusion using fluorescence correlation spectroscopy and differential dynamic microscopy within concentrated polymer solutions

    Shokeen, Namita; Issa, Christopher; Mukhopadhyay, Ashis

    2017-12-01

    We studied the diffusion of nanoparticles (NPs) within aqueous entangled solutions of polyethylene oxide (PEO) by using two different optical techniques. Fluorescence correlation spectroscopy, a method widely used to investigate nanoparticle dynamics in polymer solution, was used to measure the long-time diffusion coefficient (D) of 25 nm radius particles within high molecular weight, Mw = 600 kg/mol PEO in water solutions. Differential dynamic microscopy (DDM) was used to determine the wave-vector dependent dynamics of NPs within the same polymer solutions. Our results showed good agreement between the two methods, including demonstration of normal diffusion and almost identical diffusion coefficients obtained by both techniques. The research extends the scope of DDM to study the dynamics and rheological properties of soft matter at a nanoscale. The measured diffusion coefficients followed a scaling theory, which can be explained by the coupling between polymer dynamics and NP motion.

  14. Fluorescence Correlation Spectroscopy to Study Diffusion of Polymer Chains within Layered Hydrogen-Bonded Polymer Films

    Pristinski, Denis; Kharlampieva, Evguenia; Sukhishvili, Svetlana

    2002-03-01

    Fluorescence Correlation Spectroscopy (FCS) has been used to probe molecular motions within polymer multilayers formed by hydrogen-bonding sequential self-assembly. Polyethylene glycol (PEG) molecules were end-labeled with the fluorescent tags, and self-assembled with polymethacrylic acid (PMAA) using layer-by-layer deposition. We have found that molecules included in the top adsorbed layer have significant mobility at the millisecond time scale, probably due to translational diffusion. However, their dynamics deviate from classical Brownian motion with a single diffusion time. Possible reasons for the deviation are discussed. We found that motions were significantly slowed with increasing depth within the PEG/PMAA multilayer. This phenomena occured in a narrow pH range around 4.0 in which intermolecular interactions were relatively weak.

  15. Confined detection volume of fluorescence correlation spectroscopy by bare fiber probes.

    Lu, Guowei; Lei, Franck H; Angiboust, Jean-François; Manfait, Michel

    2010-04-01

    A fiber-tip-based near-field fluorescence correlation spectroscopy (FCS) has been developed for confining the detection volume to sub-diffraction-limited dimensions. This near-field FCS is based on near-field illumination by coupling a scanning near-field optical microscope (SNOM) to a conventional confocal FCS. Single-molecule FCS analysis at 100 nM Rhodamine 6G has been achieved by using bare chemically etched, tapered fiber tips. The detection volume under control of the SNOM system has been reduced over one order of magnitude compared to that of the conventional confocal FCS. Related factors influencing the near-field FCS performance are investigated and discussed in detail. In this proof-of-principle study, the preliminary experimental results suggest that the fiber-tip-based near-field FCS might be a good alternative to realize localized analysis at the single-molecule level.

  16. Sizes of water-soluble luminescent quantum dots measured by fluorescence correlation spectroscopy

    Zhang Pudun; Li Liang; Dong Chaoqing; Qian Huifeng; Ren Jicun

    2005-01-01

    In this paper, fluorescence correlation spectroscopy (FCS) was applied to measure the size of water-soluble quantum dots (QDs). The measurements were performed on a home-built FCS system based on the Stokes-Einstein equation. The obtained results showed that for bare CdTe QDs the sizes from FCS were larger than the ones from transmission electron microscopy (TEM). The brightness of QDs was also evaluated using FCS technique. It was found that the stability of the surface chemistry of QDs would be significantly improved by capping it with hard-core shell. Our data demonstrated that FCS is a simple, fast, and effective method for characterizing the fluorescent quantum dots, and is especially suitable for determining the fluorescent nanoparticles less than 10 nm in water solution

  17. Effect of electron correlation on the forced electric dipole transition probabilities in fsup(N) systems

    Jankowski, K.; Smentek-Mielczarek, L.

    1981-01-01

    Results of model studies of the impact of electron correlation on the forced electric dipole transition probabilities between states of the 4fsup(N) configuration are reported for the [ 3 P] 0 - [ 3 F] 4 , [ 3 H] 4 transitions in Pr 3+ : LaCl 3 and for [ 7 F] 0 - [ 5 D] 2 , [ 7 F] 1 - [ 5 D] 1 hypersensitive transitions in Eu 3+ : LaCl 3 . For the former system the correlation effects cause a modification of earlier results by 40-95 per cent, whereas for the latter the probability changes by as much as two orders of magnitude. The great changes found in the case of hypersensitive transitions suggest that electron correlation effects may belong to the most important factors determining the nature of these transitions. Several types of effective correlation operators are considered and their relative importance is discussed. The results indicate that intermediate configurations including g orbitals are very important for the description of correlation effects. (author)

  18. Correlating yeast cell stress physiology to changes in the cell surface morphology: atomic force microscopic studies.

    Canetta, Elisabetta; Walker, Graeme M; Adya, Ashok K

    2006-07-06

    Atomic Force Microscopy (AFM) has emerged as a powerful biophysical tool in biotechnology and medicine to investigate the morphological, physical, and mechanical properties of yeasts and other biological systems. However, properties such as, yeasts' response to environmental stresses, metabolic activities of pathogenic yeasts, cell-cell/cell-substrate adhesion, and cell-flocculation have rarely been investigated so far by using biophysical tools. Our recent results obtained by AFM on one strain each of Saccharomyces cerevisiae and Schizosaccharomyces pombe show a clear correlation between the physiology of environmentally stressed yeasts and the changes in their surface morphology. The future directions of the AFM related techniques in relation to yeasts are also discussed.

  19. Exploring the energy landscape of biopolymers using single molecule force spectroscopy and molecular simulations

    Hyeon, Changbong

    2010-01-01

    In recent years, single molecule force techniques have opened a new avenue to decipher the folding landscapes of biopolymers by allowing us to watch and manipulate the dynamics of individual proteins and nucleic acids. In single molecule force experiments, quantitative analyses of measurements employing sound theoretical models and molecular simulations play central role more than any other field. With a brief description of basic theories for force mechanics and molecular simulation techniqu...

  20. Characterization of protein adsorption onto FePt nanoparticles using dual-focus fluorescence correlation spectroscopy

    Pauline Maffre

    2011-07-01

    Full Text Available Using dual-focus fluorescence correlation spectroscopy, we have analyzed the adsorption of three human blood serum proteins, namely serum albumin, apolipoprotein A-I and apolipoprotein E4, onto polymer-coated, fluorescently labeled FePt nanoparticles (~12 nm diameter carrying negatively charged carboxyl groups on their surface. For all three proteins, a step-wise increase in hydrodynamic radius with protein concentration was observed, strongly suggesting the formation of protein monolayers that enclose the nanoparticles. Consistent with this interpretation, the absolute increase in hydrodynamic radius can be correlated with the molecular shapes of the proteins known from X-ray crystallography and solution experiments, indicating that the proteins bind on the nanoparticles in specific orientations. The equilibrium dissociation coefficients, measuring the affinity of the proteins to the nanoparticles, were observed to differ by almost four orders of magnitude. These variations can be understood in terms of the electrostatic properties of the proteins. From structure-based calculations of the surface potentials, positively charged patches of different extents can be revealed, through which the proteins interact electrostatically with the negatively charged nanoparticle surfaces.

  1. Effects of surface topography on SERS response: Correlating nanoscopy with spectroscopy

    Das, Sumit Kumar; Ghosh, Manash; Chowdhury, Joydeep

    2018-05-01

    This paper reports for the first time the hidden correlation between the topographical features of the bilayer Langmuir-Blodgett (LB) film substrates of stearic acid (SA) incubated in Au@Ag nanocolloids over various dipping times (DTs) with their corresponding SERS responses. The topographies of the as prepared substrates are investigated from the statistical considerations in terms of lateral correlation length, interface width, Hurst and Lyapnov exponents. The real space of the substrates are mapped directly from the FESEM and AFM images of the bilayer LB film of SA immersed in Au@Ag nanocolloids over various DTs ranging between 6 and 72 h. The SERS spectra of the Rhodamine 6G molecules adsorbed on the as prepared substrates have been reported. The statistical parameters of the substrates that exhibit maximum SERS efficacy have been suggested. The far field distributions in presence and in absence of Raman dipole together with spatial distribution of the near field from the hottest spot of the as prepared substrate have also been reported. To our knowledge, this is the first report that links nanoscopy with SERS spectroscopy from statistical considerations and is expected to open a new window towards the fabrication of more efficient and reproducible SERS active substrates in future endeavours.

  2. Determination of Dynamics of Plant Plasma Membrane Proteins with Fluorescence Recovery and Raster Image Correlation Spectroscopy.

    Laňková, Martina; Humpolíčková, Jana; Vosolsobě, Stanislav; Cit, Zdeněk; Lacek, Jozef; Čovan, Martin; Čovanová, Milada; Hof, Martin; Petrášek, Jan

    2016-04-01

    A number of fluorescence microscopy techniques are described to study dynamics of fluorescently labeled proteins, lipids, nucleic acids, and whole organelles. However, for studies of plant plasma membrane (PM) proteins, the number of these techniques is still limited because of the high complexity of processes that determine the dynamics of PM proteins and the existence of cell wall. Here, we report on the usage of raster image correlation spectroscopy (RICS) for studies of integral PM proteins in suspension-cultured tobacco cells and show its potential in comparison with the more widely used fluorescence recovery after photobleaching method. For RICS, a set of microscopy images is obtained by single-photon confocal laser scanning microscopy (CLSM). Fluorescence fluctuations are subsequently correlated between individual pixels and the information on protein mobility are extracted using a model that considers processes generating the fluctuations such as diffusion and chemical binding reactions. As we show here using an example of two integral PM transporters of the plant hormone auxin, RICS uncovered their distinct short-distance lateral mobility within the PM that is dependent on cytoskeleton and sterol composition of the PM. RICS, which is routinely accessible on modern CLSM instruments, thus represents a valuable approach for studies of dynamics of PM proteins in plants.

  3. Monitoring the staling of wheat bread using 2D MIR-NIR correlation spectroscopy

    Ringsted, Tine; Siesler, Heinz Wilhelm; Engelsen, Søren Balling

    2017-01-01

    Staling of bread is a major source of food waste and efficient monitoring of it can help the food industry in the development of anti-staling recipes. While the staling fingerprint in the mid-infrared region is fairly well established this paper set out to find the most informative parts...... of the near-infrared spectra with respect to staling. For this purpose, two-dimensional correlation spectroscopy on near- and mid-infrared spectra of wheat bread crumb during aging was employed for the first time. The important mid-infrared absorption band at 1047 cm−1 related to amylopectin retrogradation...... was found to correlate positively with increased bread hardness and to co-vary with the near-infrared band at 910 nm in the short wavelength region (r2 = 0.88 to hardness), the near-infrared band at 1688 nm in the 1. overtone region (r2 = 0.97 to hardness) and to the near-infrared band in the long...

  4. A Correlation for Forced Convective Boiling Heat Transfer of Refrigerants in a Microfin Tube

    Momoki, Satoru; Yu, Jian; Koyama, Shigeru; Fujii, Tetsu; Honda, Hiroshi

    The experimental study is reported on the forced convective boiling of pure refrigerants HCFC22, HFC134a and HCFC123 flowing in a horizontal microfin tube. The local heat transfer coefficient defined based on the actual inside surface area is measured in the ranges of mass velocity of 200 to 400 kg/m2s, heat flux of 5 to 64 kW/m2 and reduced pressure of 0.07 to 0.24. Using the Chen-type model, a new correlation for microfin tubes is proposed considering the enhancement effect of microfins on both the convective heat transfer and the nucleate boiling components. In the convective heat transfer component, the correlation to predict the heat transfer coefficient of liquid-only flow is determined from preliminary experiments on single-phase flow in microfin tubes, and the two-phase flow enhancement factor is determined from the present experimental data. For the nucleate boiling component, the correlation of Takamatsu et al. for smooth tube is modified. The prediction of the present correlation agrees well with present experimental data, and is available for several microfin tubes which were tested by other researchers.

  5. Micropillar displacements by cell traction forces are mechanically correlated with nuclear dynamics

    Li, Qingsen; Makhija, Ekta; Hameed, F.M. [Mechanobiology Institute, National University of Singapore (Singapore); Shivashankar, G.V., E-mail: shiva.gvs@gmail.com [Mechanobiology Institute, National University of Singapore (Singapore); Department of Biological Sciences, National University of Singapore (Singapore)

    2015-05-29

    Cells sense physical cues at the level of focal adhesions and transduce them to the nucleus by biochemical and mechanical pathways. While the molecular intermediates in the mechanical links have been well studied, their dynamic coupling is poorly understood. In this study, fibroblast cells were adhered to micropillar arrays to probe correlations in the physical coupling between focal adhesions and nucleus. For this, we used novel imaging setup to simultaneously visualize micropillar deflections and EGFP labeled chromatin structure at high spatial and temporal resolution. We observed that micropillar deflections, depending on their relative positions, were positively or negatively correlated to nuclear and heterochromatin movements. Our results measuring the time scales between micropillar deflections and nucleus centroid displacement are suggestive of a strong elastic coupling that mediates differential force transmission to the nucleus. - Highlights: • Correlation between focal adhesions and nucleus studied using novel imaging setup. • Micropillar and nuclear displacements were measured at high resolution. • Correlation timescales show strong elastic coupling between cell edge and nucleus.

  6. CORRELATION BETWEEN COORDINATION AND PERSONALITY TRAITS OF SOLDIERS IN BATTLE UNIT OF SLOVENIAN ARMED FORCES

    Maja Pori

    2009-11-01

    Full Text Available The aim of the study was to investigate the correlation between motor ability of coordination and personality traits of Slovenian soldiers. The subject sample consisted of 94 soldiers in a battle unit of Slovenian Armed Forces (SAF who were serving in the first brigade (age 26,5 ± 3,4 years. Motor ability of coordination was assessed with two motor tasks (polygon bac- kwards and figure 8 duck test. The structure of personality traits was measured with a FPI (Freiburg Personality Inventory included 114 items and measured 9 personality traits of order I (neuroticism, impulsivity, depression, irritability, sociability, calmness, dominance, suppression, sincerity and 3 personality traits of order II (extroversion, emotional istability, masculinity. The correlation between coordination and personality traits was estimated by the Pearson’s correlation coefficient. The results show that soldiers who did worse in motor test polygon backwards were more neurotic, suppressed, and impulsive. They also tend to be more sociable. The correlation between second test of coordination shows that better soldiers in this test are more extrovert or less introvert.

  7. Chemical Force Spectroscopy Evidence Supporting the Layer-by-Layer Model of Organic Matter Binding to Iron (oxy)Hydroxide Mineral Surfaces

    Chassé , Alexander W.; Ohno, Tsutomu; Higgins, Steven R.; Amirbahman, Aria; Yildirim, Nadir; Parr, Thomas B.

    2015-01-01

    © 2015 American Chemical Society. The adsorption of dissolved organic matter (DOM) to metal (oxy)hydroxide mineral surfaces is a critical step for C sequestration in soils. Although equilibrium studies have described some of the factors controlling this process, the molecular-scale description of the adsorption process has been more limited. Chemical force spectroscopy revealed differing adhesion strengths of DOM extracted from three soils and a reference peat soil material to an iron (oxy)hydroxide mineral surface. The DOM was characterized using ultrahigh-resolution negative ion mode electrospray ionization Fourier Transform ion cyclotron resonance mass spectrometry. The results indicate that carboxyl-rich aromatic and N-containing aliphatic molecules of DOM are correlated with high adhesion forces. Increasing molecular mass was shown to decrease the adhesion force between the mineral surface and the DOM. Kendrick mass defect analysis suggests that mechanisms involving two carboxyl groups result in the most stable bond to the mineral surface. We conceptualize these results using a layer-by-layer "onion" model of organic matter stabilization on soil mineral surfaces.

  8. Chemical Force Spectroscopy Evidence Supporting the Layer-by-Layer Model of Organic Matter Binding to Iron (oxy)Hydroxide Mineral Surfaces

    Chassé, Alexander W.

    2015-08-18

    © 2015 American Chemical Society. The adsorption of dissolved organic matter (DOM) to metal (oxy)hydroxide mineral surfaces is a critical step for C sequestration in soils. Although equilibrium studies have described some of the factors controlling this process, the molecular-scale description of the adsorption process has been more limited. Chemical force spectroscopy revealed differing adhesion strengths of DOM extracted from three soils and a reference peat soil material to an iron (oxy)hydroxide mineral surface. The DOM was characterized using ultrahigh-resolution negative ion mode electrospray ionization Fourier Transform ion cyclotron resonance mass spectrometry. The results indicate that carboxyl-rich aromatic and N-containing aliphatic molecules of DOM are correlated with high adhesion forces. Increasing molecular mass was shown to decrease the adhesion force between the mineral surface and the DOM. Kendrick mass defect analysis suggests that mechanisms involving two carboxyl groups result in the most stable bond to the mineral surface. We conceptualize these results using a layer-by-layer "onion" model of organic matter stabilization on soil mineral surfaces.

  9. The interaction of 2-mercaptobenzimidazole with human serum albumin as determined by spectroscopy, atomic force microscopy and molecular modeling.

    Li, Yuqin; Jia, Baoxiu; Wang, Hao; Li, Nana; Chen, Gaopan; Lin, Yuejuan; Gao, Wenhua

    2013-04-01

    The interaction of 2-mercaptobenzimidazole (MBI) with human serum albumin (HSA) was studied in vitro by equilibrium dialysis under normal physiological conditions. This study used fluorescence, ultraviolet-visible spectroscopy (UV-vis), Fourier transform infrared (FT-IR), circular dichroism (CD) and Raman spectroscopy, atomic force microscopy (AFM) and molecular modeling techniques. Association constants, the number of binding sites and basic thermodynamic parameters were used to investigate the quenching mechanism. Based on the fluorescence resonance energy transfer, the distance between the HSA and MBI was 2.495 nm. The ΔG(0), ΔH(0), and ΔS(0) values across temperature indicated that the hydrophobic interaction was the predominant binding Force. The UV, FT-IR, CD and Raman spectra confirmed that the HSA secondary structure was altered in the presence of MBI. In addition, the molecular modeling showed that the MBI-HSA complex was stabilized by hydrophobic forces, which resulted from amino acid residues. The AFM results revealed that the individual HSA molecule dimensions were larger after interaction with MBI. Overall, this study suggested a method for characterizing the weak intermolecular interaction. In addition, this method is potentially useful for elucidating the toxigenicity of MBI when it is combined with the biomolecular function effect, transmembrane transport, toxicological testing and other experiments. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. spectroscopy

    Aghomotsegin

    2015-10-14

    Oct 14, 2015 ... characterized by using phenotypic, API and Fourier transform infrared (FTIR) spectroscopy methods. One hundred and fifty-seven (157) strains were isolated from 13 cheese samples, and identification test was performed for 83 strains. At the end of the study, a total of 22 Lactococcus sp., 36 Enterecoccus ...

  11. Neural correlates of belief-bias reasoning under time pressure: a near-infrared spectroscopy study.

    Tsujii, Takeo; Watanabe, Shigeru

    2010-04-15

    The dual-process theory of reasoning explained the belief-bias effect, the tendency for human reasoning to be erroneously biased when logical conclusions are incongruent with belief about the world, by proposing a belief-based fast heuristic system and a logic-based slow analytic system. Although the claims were supported by behavioral findings that the belief-bias effect was enhanced when subjects were not given sufficient time for reasoning, the neural correlates were still unknown. The present study therefore examined the relationship between the time-pressure effect and activity in the inferior frontal cortex (IFC) during belief-bias reasoning using near-infrared spectroscopy (NIRS). Forty-eight subjects performed congruent and incongruent reasoning tasks, involving long-span (20 s) and short-span trials (10 s). Behavioral analysis found that only incongruent reasoning performance was impaired by the time-pressure of short-span trials. NIRS analysis found that the time-pressure decreased right IFC activity during incongruent trials. Correlation analysis showed that subjects with enhanced right IFC activity could perform better in incongruent trials, while subjects for whom the right IFC activity was impaired by the time-pressure could not maintain better reasoning performance. These findings suggest that the right IFC may be responsible for the time-pressure effect in conflicting reasoning processes. When the right IFC activity was impaired in the short-span trials in which subjects were not given sufficient time for reasoning, the subjects may rely on the fast heuristic system, which result in belief-bias responses. We therefore offer the first demonstration of neural correlates of time-pressure effect on the IFC activity in belief-bias reasoning. Copyright 2009 Elsevier Inc. All rights reserved.

  12. MR spectroscopy of prostate cancer: correlation study of metabolic characters with Gleason score

    Wang Xiaoying; Li Feiyu; Jiang Xuexiang; Shan Gangzhi; Zhou Liangping; Ding Jianping

    2006-01-01

    Objective: Using magnetic resonance spectroscopy (MRS) to measure the metabolic ratio [(choline + creatine)/citrate, CC/C] of prostate cancer(PCa) and to probe the correlation between the value of CC/C ratio and Gleason score. Methods: Twenty-one cases of PCa proved by operation or systemic biopsy were examined by MRS. The prostate was divided into 6 regions (left/right bottom, middle and tip), and the CC/C value of each region was measured. After biopsy, all the puncture locations were marked and enrolled in one of the regions mentioned above and the Gleason scores were recorded. The ratio of CC/C measured by MRS was compared with Gleason score in the corresponding regions. Results: The average CC/C ratio of the 74 regions with PCa was 2.13±0.82, whereas the average CC/C ratio of the 52 regions without PCa was 0.59±0.20. The difference of CC/C value was statistically significant (t=7.72, P=0.00). The ratios of CC/C in the regions of PCa were correlated with Gleason score (r=0.659, P=0.01). In group 1 (Gleason score≥7), the average CC/C ratio was 2.61±0.79. However, in group 2 (Gleasonscore <7) the average CC/C ratio was 1.69±0.59. There was statistically difference between the two groups (t=3.06, P=0.01). Conclusion: The metabolic ratio of CC/C is correlated to the Gleason score of PCa. MRS may be a useful method to evaluate the malignancy of PCa noninvasively. (authors)

  13. Analysis of Chuanxiong Rhizoma and its active components by Fourier transform infrared spectroscopy combined with two-dimensional correlation infrared spectroscopy.

    Guo, Yizhen; Lv, Beiran; Wang, Jingjuan; Liu, Yang; Sun, Suqin; Xiao, Yao; Lu, Lina; Xiang, Li; Yang, Yanfang; Qu, Lei; Meng, Qinghong

    2016-01-15

    As complicated mixture systems, active components of Chuanxiong Rhizoma are very difficult to identify and discriminate. In this paper, the macroscopic IR fingerprint method including Fourier transform infrared spectroscopy (FT-IR), the second derivative infrared spectroscopy (SD-IR) and two-dimensional correlation infrared spectroscopy (2DCOS-IR), was applied to study and identify Chuanxiong raw materials and its different segmented production of HPD-100 macroporous resin. Chuanxiong Rhizoma is rich in sucrose. In the FT-IR spectra, water eluate is more similar to sucrose than the powder and the decoction. Their second derivative spectra amplified the differences and revealed the potentially characteristic IR absorption bands and combined with the correlation coefficient, concluding that 50% ethanol eluate had more ligustilide than other eluates. Finally, it can be found from 2DCOS-IR spectra that proteins were extracted by ethanol from Chuanxiong decoction by HPD-100 macroporous resin. It was demonstrated that the above three-step infrared spectroscopy could be applicable for quick, non-destructive and effective analysis and identification of very complicated and similar mixture systems of traditional Chinese medicines. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Rapid identification of Pterocarpus santalinus and Dalbergia louvelii by FTIR and 2D correlation IR spectroscopy

    Zhang, Fang-Da; Xu, Chang-Hua; Li, Ming-Yu; Huang, An-Min; Sun, Su-Qin

    2014-07-01

    Since Pterocarpus santalinus and Dalbergia louvelii, which are of precious Rosewood, are very similar in their appearance and anatomy characteristics, cheaper Hongmu D. louvelii is often illegally used to impersonate valuable P. santalinus, especially in Chinese furniture manufacture. In order to develop a rapid and effective method for easy confused wood furniture differentiation, we applied tri-step identification method, i.e., conventional infrared spectroscopy (FT-IR), second derivative infrared (SD-IR) spectroscopy and two-dimensional correlation infrared (2DCOS-IR) spectroscopy to investigate P. santalinus and D. louvelii furniture. According to FT-IR and SD-IR spectra, it has been found two unconditional stable difference at 848 cm-1 and 700 cm-1 and relative stable differences at 1735 cm-1, 1623 cm-1, 1614 cm-1, 1602 cm-1, 1509 cm-1, 1456 cm-1, 1200 cm-1, 1158 cm-1, 1055 cm-1, 1034 cm-1 and 895 cm-1 between D. louvelii and P. santalinus IR spectra. The stable discrepancy indicates that the category of extractives is different between the two species. Besides, the relative stable differences imply that the content of holocellulose in P. santalinus is more than that of D. louvelii, whereas the quantity of extractives in D. louvelii is higher. Furthermore, evident differences have been observed in their 2DCOS-IR spectra of 1550-1415 cm-1 and 1325-1030 cm-1. P. santalinus has two strong auto-peaks at 1459 cm-1 and 1467 cm-1, three mid-strong auto-peaks at 1518 cm-1, 1089 cm-1 and 1100 cm-1 and five weak auto-peaks at 1432 cm-1, 1437 cm-1, 1046 cm-1, 1056 cm-1 and 1307 cm-1 while D. louvelii has four strong auto-peaks at 1465 cm-1, 1523 cm-1, 1084 cm-1 and 1100 cm-1, four mid-strong auto-peaks at 1430 cm-1, 1499 cm-1, 1505 cm-1 and 1056 cm-1 and two auto-peaks at 1540 cm-1 and 1284 cm-1. This study has proved that FT-IR integrated with 2DCOS-IR could be applicable for precious wood furniture authentication in a direct, rapid and holistic manner.

  15. Chemical profiling and adulteration screening of Aquilariae Lignum Resinatum by Fourier transform infrared (FT-IR) spectroscopy and two-dimensional correlation infrared (2D-IR) spectroscopy.

    Qu, Lei; Chen, Jian-Bo; Zhang, Gui-Jun; Sun, Su-Qin; Zheng, Jing

    2017-03-05

    As a kind of expensive perfume and valuable herb, Aquilariae Lignum Resinatum (ALR) is often adulterated for economic motivations. In this research, Fourier transform infrared (FT-IR) spectroscopy is employed to establish a simple and quick method for the adulteration screening of ALR. First, the principal chemical constituents of ALR are characterized by FT-IR spectroscopy at room temperature and two-dimensional correlation infrared (2D-IR) spectroscopy with thermal perturbation. Besides the common cellulose and lignin compounds, a certain amount of resin is the characteristic constituent of ALR. Synchronous and asynchronous 2D-IR spectra indicate that the resin (an unstable secondary metabolite) is more sensitive than cellulose and lignin (stable structural constituents) to the thermal perturbation. Using a certified ALR sample as the reference, the infrared spectral correlation threshold is determined by 30 authentic samples and 6 adulterated samples. The spectral correlation coefficient of an authentic ALR sample to the standard reference should be not less than 0.9886 (p=0.01). Three commercial adulterated ALR samples are identified by the correlation threshold. Further interpretation of the infrared spectra of the adulterated samples indicates the common adulterating methods - counterfeiting with other kind of wood, adding ingredient such as sand to increase the weight, and adding the cheap resin such as rosin to increase the content of resin compounds. Results of this research prove that FT-IR spectroscopy can be used as a simple and accurate quality control method of ALR. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Characterization of gold nanoparticle films: Rutherford backscattering spectroscopy, scanning electron microscopy with image analysis, and atomic force microscopy

    Pia C. Lansåker

    2014-10-01

    Full Text Available Gold nanoparticle films are of interest in several branches of science and technology, and accurate sample characterization is needed but technically demanding. We prepared such films by DC magnetron sputtering and recorded their mass thickness by Rutherford backscattering spectroscopy. The geometric thickness dg—from the substrate to the tops of the nanoparticles—was obtained by scanning electron microscopy (SEM combined with image analysis as well as by atomic force microscopy (AFM. The various techniques yielded an internally consistent characterization of the films. In particular, very similar results for dg were obtained by SEM with image analysis and by AFM.

  17. Correlation Networks from Flows. The Case of Forced and Time-Dependent Advection-Diffusion Dynamics.

    Liubov Tupikina

    Full Text Available Complex network theory provides an elegant and powerful framework to statistically investigate different types of systems such as society, brain or the structure of local and long-range dynamical interrelationships in the climate system. Network links in climate networks typically imply information, mass or energy exchange. However, the specific connection between oceanic or atmospheric flows and the climate network's structure is still unclear. We propose a theoretical approach for verifying relations between the correlation matrix and the climate network measures, generalizing previous studies and overcoming the restriction to stationary flows. Our methods are developed for correlations of a scalar quantity (temperature, for example which satisfies an advection-diffusion dynamics in the presence of forcing and dissipation. Our approach reveals that correlation networks are not sensitive to steady sources and sinks and the profound impact of the signal decay rate on the network topology. We illustrate our results with calculations of degree and clustering for a meandering flow resembling a geophysical ocean jet.

  18. Ground reaction force analysed with correlation coefficient matrix in group of stroke patients.

    Szczerbik, Ewa; Krawczyk, Maciej; Syczewska, Małgorzata

    2014-01-01

    Stroke is the third cause of death in contemporary society and causes many disorders. Clinical scales, ground reaction force (GRF) and objective gait analysis are used for assessment of patient's rehabilitation progress during treatment. The goal of this paper is to assess whether signal correlation coefficient matrix applied to GRF can be used for evaluation of the status of post-stroke patients. A group of patients underwent clinical assessment and instrumented gait analysis simultaneously three times. The difference between components of patient's GRF (vertical, fore/aft, med/lat) and normal ones (reference GRF of healthy subjects) was calculated as correlation coefficient. Patients were divided into two groups ("worse" and "better") based on the clinical functional scale tests done at the beginning of rehabilitation process. The results obtained by these two groups were compared using statistical analysis. An increase of median value of correlation coefficient is observed in all components of GRF, but only in non-paretic leg. Analysis of GRF signal can be helpful in assessment of post-stroke patients during rehabilitation. Improvement in stroke patients was observed in non-paretic leg of the "worse" group. GRF analysis should not be the only tool for objective validation of patient's improvement, but could be used as additional source of information.

  19. Interaction of an anticancer peptide fragment of azurin with p53 and its isolated domains studied by atomic force spectroscopy.

    Bizzarri, Anna Rita; Santini, Simona; Coppari, Emilia; Bucciantini, Monica; Di Agostino, Silvia; Yamada, Tohru; Beattie, Craig W; Cannistraro, Salvatore

    2011-01-01

    p28 is a 28-amino acid peptide fragment of the cupredoxin azurin derived from Pseudomonas aeruginosa that preferentially penetrates cancerous cells and arrests their proliferation in vitro and in vivo. Its antitumor activity reportedly arises from post-translational stabilization of the tumor suppressor p53 normally downregulated by the binding of several ubiquitin ligases. This would require p28 to specifically bind to p53 to inhibit specific ligases from initiating proteosome-mediated degradation. In this study, atomic force spectroscopy, a nanotechnological approach, was used to investigate the interaction of p28 with full-length p53 and its isolated domains at the single molecule level. Analysis of the unbinding forces and the dissociation rate constant suggest that p28 forms a stable complex with the DNA-binding domain of p53, inhibiting the binding of ubiquitin ligases other than Mdm2 to reduce proteasomal degradation of p53.

  20. Linear correlation for identification of materials by laser induced breakdown spectroscopy: Improvement via spectral filtering and masking

    Gornushkin, I.B., E-mail: igor.gornushkin@bam.d [BAM Federal Institute for Materials Research and Testing, Berlin (Germany); Panne, U. [BAM Federal Institute for Materials Research and Testing, Berlin (Germany); Winefordner, J.D. [University of Florida, Gainesville, Florida (United States)

    2009-10-15

    The purpose of this work is to improve the performance of a linear correlation method used for material identification in laser induced breakdown spectroscopy. The improved correlation procedure is proposed based on the selection and use of only essential spectral information and ignoring empty spectral fragments. The method is tested on glass samples of forensic interest. The 100% identification capability of the new method is demonstrated in contrast to the traditional approach where the identification rate falls below 100% for many samples.

  1. Three-dimensional scanning force/tunneling spectroscopy at room temperature

    Sugimoto, Yoshiaki; Ueda, Keiichi; Abe, Masayuki; Morita, Seizo

    2012-01-01

    We simultaneously measured the force and tunneling current in three-dimensional (3D) space on the Si(111)-(7 × 7) surface using scanning force/tunneling microscopy at room temperature. The observables, the frequency shift and the time-averaged tunneling current were converted to the physical quantities of interest, i.e. the interaction force and the instantaneous tunneling current. Using the same tip, the local density of states (LDOS) was mapped on the same surface area at constant height by measuring the time-averaged tunneling current as a function of the bias voltage at every lateral position. LDOS images at negative sample voltages indicate that the tip apex is covered with Si atoms, which is consistent with the Si-Si covalent bonding mechanism for AFM imaging. A measurement technique for 3D force/current mapping and LDOS imaging on the equivalent surface area using the same tip was thus demonstrated. (paper)

  2. Single-molecule force-conductance spectroscopy of hydrogen-bonded complexes

    Pirrotta, Alessandro; De Vico, Luca; Solomon, Gemma C.

    2017-01-01

    to inform about molecular recognition events at the single-molecule limit. For this, we consider the force-conductance characteristics of a prototypical class of hydrogen bonded bimolecular complexes sandwiched between gold electrodes. The complexes consist of derivatives of a barbituric acid and a Hamilton...... is mechanically manipulated. The implication is that force and conductance provide complementary information about the evolution of molecules in junctions that can be used to interrogate basic structure-transport relations at the single-molecule limit....

  3. Diagnosis of non-exudative (DRY) age related macular degeneration by non-invasive photon-correlation spectroscopy.

    Fankhauser, Franz Ii; Ott, Maria; Munteanu, Mihnea

    2016-01-01

    Photon-correlation spectroscopy (PCS) (quasi-elastic light scattering spectroscopy, dynamic light scattering spectroscopy) allows the non-invasively reveal of local dynamics and local heterogeneities of macromolecular systems. The capability of this technique to diagnose the retinal pathologies by in-vivo investigations of spatial anomalies of retinas displaying non-exudative senile macular degeneration was evaluated. Further, the potential use of the technique for the diagnosis of the macular degeneration was analyzed and displayed by the Receiver Operating Curve (ROC). The maculae and the peripheral retina of 73 normal eyes and of 26 eyes afflicted by an early stage of non-exudative senile macular degeneration were characterized by time-correlation functions and analyzed in terms of characteristic decay times and apparent size distributions. The characteristics of the obtained time-correlation functions of the eyes afflicted with nonexudative macular degeneration and of normal eyes differed significantly, which could be referred to a significant change of the nano- and microstructure of the investigated pathologic maculas. Photon-correlation spectroscopy is able to assess the macromolecular and microstructural aberrations in the macula afflicted by non-exudative, senile macular degeneration. It has been demonstrated that macromolecules of this disease show a characteristic abnormal behavior in the macula.

  4. a Study of the Concentration Dependence of Macromolecular Diffusion Using Photon Correlation Spectroscopy.

    Marlowe, Robert Lloyd

    The dynamic light scattering technique of photon correlation spectroscopy has been used to investigate the dependence of the mutual diffusion coefficient of a macromolecular system upon concentration. The first part of the research was devoted to the design and construction of a single-clipping autocorrelator based on newly-developed integrated circuits. The resulting 128 channel instrument can perform real time autocorrelation for sample time intervals >(, )10 (mu)s, and batch processed autocorrelation for intervals down to 3 (mu)s. An improved design for a newer, all-digital autocorrelator is given. Homodyne light scattering experiments were then undertaken on monodisperse solutions of polystyrene spheres. The single-mode TEM(,oo) beam of an argon-ion laser ((lamda) = 5145 (ANGSTROM)) was used as the light source; all solutions were studied at room temperature. The scattering angle was varied from 30(DEGREES) to 110(DEGREES). Excellent agreement with the manufacturer's specification for the particle size was obtained from the photon correlation studies. Finally, aqueous solutions of the globular protein ovalbumin, ranging in concentration from 18.9 to 244.3 mg/ml, were illuminated under the same conditions of temperature and wavelength as before; the homodyne scattered light was detected at a fixed scattering angle of 30(DEGREES). The single-clipped photocount autocorrelation function was analyzed using the homodyne exponential integral method of Meneely et al. The resulting diffusion coefficients showed a general linear dependence upon concentration, as predicted by the generalized Stokes-Einstein equation. However, a clear peak in the data was evident at c (TURNEQ) 100 mg/ml, which could not be explained on the basis of a non -interacting particle theory. A semi-quantitative approach based on the Debye-Huckel theory of electrostatic interactions is suggested as the probable cause for the peak's rise, and an excluded volume effect for its decline.

  5. Using fluorescence correlation spectroscopy to study conformational changes in denatured proteins.

    Sherman, Eilon; Itkin, Anna; Kuttner, Yosef Yehuda; Rhoades, Elizabeth; Amir, Dan; Haas, Elisha; Haran, Gilad

    2008-06-01

    Fluorescence correlation spectroscopy (FCS) is a sensitive analytical tool that allows dynamics and hydrodynamics of biomolecules to be studied under a broad range of experimental conditions. One application of FCS of current interest is the determination of the size of protein molecules in the various states they sample along their folding reaction coordinate, which can be accessed through the measurement of diffusion coefficients. It has been pointed out that the analysis of FCS curves is prone to artifacts that may lead to erroneous size determination. To set the stage for FCS studies of unfolded proteins, we first show that the diffusion coefficients of small molecules as well as proteins can be determined accurately even in the presence of high concentrations of co-solutes that change the solution refractive index significantly. Indeed, it is found that the Stokes-Einstein relation between the measured diffusion coefficient and solution viscosity holds even in highly concentrated glycerol or guanidinium hydrochloride (GuHCl) solutions. These measurements form the basis for an investigation of the structure of the denatured state of two proteins, the small protein L and the larger, three-domain protein adenylate kinase (AK). FCS is found useful for probing expansion in the denatured state beyond the unfolding transition. It is shown that the denatured state of protein L expands as the denaturant concentration increases, in a process akin to the transition from a globule to a coil in polymers. This process continues at least up to 5 M GuHCl. On the other hand, the denatured state of AK does not seem to expand much beyond 2 M GuHCl, a result that is in qualitative accord with single-molecule fluorescence histograms. Because both the unfolding transition and the coil-globule transition of AK occur at a much lower denaturant concentration than those of protein L, a possible correlation between the two phenomena is suggested.

  6. Wilson's disease: 31P and 1H MR spectroscopy and clinical correlation

    Sinha, Sanjib; Taly, A.B.; Prashanth, L.K.; Ravishankar, S.; Vasudev, M.K.

    2010-01-01

    Proton ( 1 H) magnetic resonance spectroscopy (MRS) changes are noted in Wilson's disease (WD). However, there are no studies regarding membrane phospholipid abnormality using 31 P MRS in these patients. We aimed to analyze the striatal spectroscopic abnormalities using 31 P and 1 H MRS in WD. Forty patients of WD (treated, 29; untreated,11) and 30 controls underwent routine MR image sequences and in vivo 2-D 31 P and 1 H MRS of basal ganglia using an image-selected technique on a 1.5-T MRI scanner. Statistical analysis was done using Student's t test. The mean durations of illness and treatment were 6.2 ± 7.4 and 4.8 ± 5.9 years, respectively. MRI images were abnormal in all the patients. 1 H MRS revealed statistically significant reduction of N-acetyl aspartate (NAA)/choline (Cho) and NAA/creatine ratios in striatum ( 1 H MRS) of treated patients compared to controls. The mean values of phosphomonoesters (PME) (p 31 P MRS study. The duration of illness correlated well with increased PME/PDE [p < 0.001], PME/TPh [p < 0.05], and PDE/TPh [p < 0.05] and decreased NAA/Cho [p < 0.05] ratios. There was correlation of MRI score and reduced NAA/Cho ratio with disease severity. The PME/PDE ratio (right) was elevated in the treated group [p < 0.001] compared to untreated group. There is reduced breakdown and/or increased synthesis of membrane phospholipids and increased neuronal damage in basal ganglia in patients with WD. (orig.)

  7. Investigation of hyperfine interactions in DNA nitrogenous bases using perturbed angular correlation spectroscopy

    Silva, Andreia dos Santos; Carbonari, Artur Wilson; Lapolli, Andre Luis; Saxena, Rajendra Narain; Saitovitch, Henrique

    2013-01-01

    Perturbed γγ angular correlations (PAC) spectroscopy has been used to study the DNA nitrogenous bases (adenine, cytosine, guanine, thymine), using 111 In→ 111 Cd and 111m Cd→ 111 Cd probe nuclei. One of the advantages of applying PAC technique to biological molecules is that the experiments can be carried out on molecules in aqueous solution [1], approaching the function of molecules under conditions that are close to in vivo conditions. The measurements were carried out for DNA nitrogenous bases molecules at 295 K and 77 K in order to investigate dynamic and static hyperfine interactions, respectively. The interpretation of the results was based on the measurements of dynamic interaction characterized by the decay constant from which valuable information on the macroscopic behavior of the molecules was obtained [2; 3]. On the other hand, PAC measurements at low temperature showed interaction frequency (ν Q ), asymmetry parameter (η) and the distribution of the quadrupole frequency (δ). These parameters provide a local microscopic description of the chemical environment in the neighborhood of the probe nuclei. Results showed differences in the hyperfine interactions of probe nuclei bound to the studied biomolecules. Such differences were observed by variations in the hyperfine parameters, which depended on the type of biomolecule and the results also showed that the probe nuclei bounded at the molecules in some cases and at others did not. (author)

  8. Confined diffusion in tubular structures analyzed by fluorescence correlation spectroscopy on a mirror

    Etienne, Emilien; Lenne, Pierre-Francois; Sturgis, James N.; Rigneault, Herve

    2006-01-01

    In fluorescence correlation spectroscopy (FCS) analysis it is generally assumed that molecular species diffuse freely in volumes much larger than the three-dimensional FCS observation volume. However, this standard assumption is not valid in many measurement conditions, particularly in tubular structures with diameters in the micrometer range, such as those found in living cells (organelles, dendrites) and microfluidic devices (capillaries,reaction chambers). As a result the measured autocorrelation functions (ACFs) deviate from those predicted for free diffusion, and this can shift the measured diffusion coefficient by as much as ∼50% when the tube diameter is comparable with the axial extension of the FCS observation volume. We show that the range of validity of the FCS measurements can be drastically improved if the tubular structures are located in the close vicinity of a mirror on which FCS is performed. In this case a new fluctuation time in the ACF, arising from the diffusion of fluorescent probes in optical fringes,permits measurement of the real diffusion coefficient within the tubular structure without assumptions about either the confined geometry orthe FCS observation volume geometry. We show that such a measurement can be done when the tubular structure contains at least one pair of dark and bright fringes resulting from interference between the incoming and the reflected excitation beams on the mirror surface. Measurement of the diffusion coefficient of the enhanced green fluorescent protein (EGFP) and IscS-EGFP in the cytoplasm of living Escherichiacoli illustrates the capabilities of the technique

  9. On the performance of bioanalytical fluorescence correlation spectroscopy measurements in a multiparameter photon-counting microscope

    Mazouchi, Amir; Liu Baoxu; Bahram, Abdullah [Department of Physics, Institute for Optical Sciences, University of Toronto, Toronto (Canada); Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, ON, L5L 1C6 (Canada); Gradinaru, Claudiu C., E-mail: claudiu.gradinaru@utoronto.ca [Department of Physics, Institute for Optical Sciences, University of Toronto, Toronto (Canada); Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, ON, L5L 1C6 (Canada)

    2011-02-28

    Fluorescence correlation spectroscopy (FCS) data acquisition and analysis routines were developed and implemented in a home-built, multiparameter photon-counting microscope. Laser excitation conditions were investigated for two representative fluorescent probes, Rhodamine110 and enhanced green fluorescent protein (EGFP). Reliable local concentrations and diffusion constants were obtained by fitting measured FCS curves, provided that the excitation intensity did not exceed 20% of the saturation level for each fluorophore. Accurate results were obtained from FCS measurements for sample concentrations varying from pM to {mu}M range, as well as for conditions of high background signals. These experimental constraints were found to be determined by characteristics of the detection system and by the saturation behavior of the fluorescent probes. These factors actually limit the average number of photons that can be collected from a single fluorophore passing through the detection volume. The versatility of our setup and the data analysis capabilities were tested by measuring the mobility of EGFP in the nucleus of Drosophila cells under conditions of high concentration and molecular crowding. As a bioanalytical application, we studied by FCS the binding affinity of a novel peptide-based drug to the cancer-regulating STAT3 protein and corroborated the results with fluorescence polarization analysis derived from the same photon data.

  10. Comparison of photon correlation spectroscopy with photosedimentation analysis for the determination of aqueous colloid size distributions

    Rees, Terry F.

    1990-01-01

    Colloidal materials, dispersed phases with dimensions between 0.001 and 1 μm, are potential transport media for a variety of contaminants in surface and ground water. Characterization of these colloids, and identification of the parameters that control their movement, are necessary before transport simulations can be attempted. Two techniques that can be used to determine the particle-size distribution of colloidal materials suspended in natural waters are compared. Photon correlation Spectroscopy (PCS) utilizes the Doppler frequency shift of photons scattered off particles undergoing Brownian motion to determine the size of colloids suspended in water. Photosedimentation analysis (PSA) measures the time-dependent change in optical density of a suspension of colloidal particles undergoing centrifugation. A description of both techniques, important underlying assumptions, and limitations are given. Results for a series of river water samples show that the colloid-size distribution means are statistically identical as determined by both techniques. This also is true of the mass median diameter (MMD), even though MMD values determined by PSA are consistently smaller than those determined by PCS. Because of this small negative bias, the skew parameters for the distributions are generally smaller for the PCS-determined distributions than for the PSA-determined distributions. Smaller polydispersity indices for the distributions are also determined by PCS.

  11. Photolithography and Fluorescence Correlation Spectroscopy used to examine the rates of exchange in reverse micelle systems

    Norris, Zach; Mawson, Cara; Johnson, Kyron; Kessler, Sarah; Rebecca, Anne; Wolf, Nathan; Lim, Michael; Nucci, Nathaniel

    Reverse micelles are molecular complexes that encapsulate a nanoscale pool of water in a surfactant shell dissolved in non-polar solvent. These complexes have a wide range of applications, and in all cases, the degree to which reverse micelles (RM) exchange their contents is relevant for their use. Despite its importance, this aspect of RM behavior is poorly understood. Photolithography is employed here to create micro and nano scale fluidic systems in which mixing rates can be precisely measured using fluorescence correlation spectroscopy (FCS). Micro-channel patterns are etched using reactive ion etching process into a layer of silicon dioxide on crystalline silicon substrates. Solutions containing mixtures of reverse micelles, proteins, and fluorophores are placed into reservoirs in the patterns, while diffusion and exchange between RMs is monitored using a FCS system built from a modified confocal Raman spectrometer. Using this approach, the diffusion and exchange rates for RM systems are measured as a function of the components of the RM mixture. Funding provided by Rowan University.

  12. Mapping the dynamical organization of the cell nucleus through fluorescence correlation spectroscopy.

    Stortz, Martin; Angiolini, Juan; Mocskos, Esteban; Wolosiuk, Alejandro; Pecci, Adali; Levi, Valeria

    2018-05-01

    The hierarchical organization of the cell nucleus into specialized open reservoirs and the nucleoplasm overcrowding impose restrictions to the mobility of biomolecules and their interactions with nuclear targets. These properties determine that many nuclear functions such as transcription, replication, splicing or DNA repair are regulated by complex, dynamical processes that do not follow simple rules. Advanced fluorescence microscopy tools and, in particular, fluorescence correlation spectroscopy (FCS) provide complementary and exquisite information on the dynamics of fluorescent labeled molecules moving through the nuclear space and are helping us to comprehend the complexity of the nuclear structure. Here, we describe how FCS methods can be applied to reveal the dynamical organization of the nucleus in live cells. Specifically, we provide instructions for the preparation of cellular samples with fluorescent tagged proteins and detail how FCS can be easily instrumented in commercial confocal microscopes. In addition, we describe general rules to set the parameters for one and two-color experiments and the required controls for these experiments. Finally, we review the statistical analysis of the FCS data and summarize the use of numerical simulations as a complementary approach that helps us to understand the complex matrix of molecular interactions network within the nucleus. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Evidence for atomic scale disorder in indium nitride from perturbed angular correlation spectroscopy

    Dogra, R; Shrestha, S K; Byrne, A P; Ridgway, M C; Edge, A V J; Vianden, R; Penner, J; Timmers, H

    2005-01-01

    The crystal lattice of bulk grains and state-of-the-art films of indium nitride was investigated at the atomic scale with perturbed angular correlation spectroscopy using the 111 In/Cd radioisotope probe. The probe was introduced during sample synthesis, by diffusion and by ion implantation. The mean quadrupole interaction frequency ν Q = 28 MHz was observed at the indium probe site in all types of indium nitride samples with broad frequency distributions. The observed small, but non-zero, asymmetry parameter indicates broken symmetry around the probe atoms. Results have been compared with theoretical calculations based on the point charge model. The consistency of the experimental results and their independence of the preparation technique suggest that the origin of the broad frequency distribution is inherent to indium nitride, indicating a high degree of disorder at the atomic scale. Due to the low dissociation temperature of indium nitride, furnace and rapid thermal annealing at atmospheric pressure reduce the lattice disorder only marginally

  14. Cell cycle-dependent mobility of Cdc45 determined in vivo by fluorescence correlation spectroscopy.

    Ronan Broderick

    Full Text Available Eukaryotic DNA replication is a dynamic process requiring the co-operation of specific replication proteins. We measured the mobility of eGFP-Cdc45 by Fluorescence Correlation Spectroscopy (FCS in vivo in asynchronous cells and in cells synchronized at the G1/S transition and during S phase. Our data show that eGFP-Cdc45 mobility is faster in G1/S transition compared to S phase suggesting that Cdc45 is part of larger protein complex formed in S phase. Furthermore, the size of complexes containing Cdc45 was estimated in asynchronous, G1/S and S phase-synchronized cells using gel filtration chromatography; these findings complemented the in vivo FCS data. Analysis of the mobility of eGFP-Cdc45 and the size of complexes containing Cdc45 and eGFP-Cdc45 after UVC-mediated DNA damage revealed no significant changes in diffusion rates and complex sizes using FCS and gel filtration chromatography analyses. This suggests that after UV-damage, Cdc45 is still present in a large multi-protein complex and that its mobility within living cells is consistently similar following UVC-mediated DNA damage.

  15. Study of molybdenum oxide by means of Perturbed Angular Correlations (PAC) and Mössbauer spectroscopy

    Among transition-metal oxides, the molybdenum oxide compounds are particularly attractive due to the structural (2D) anisotropy and to the ability of the molybdenum ion to change its oxidation state. These properties make it suitable for applications on, e.g., chemical sensors, solar cells, catalytic and optoelectronic devices. At ISOLDE we aim studying the incorporation of selected dopants by ion implantation, using the nuclear techniques of Perturbed Angular Correlations (PAC) and Mössbauer spectroscopy (MS). Both techniques make use of highly diluted radioactive probe nuclei, which interact – as atomic-sized tips – with the host atoms and defects. The objectives of this project are to study at the atomic scale the probe’s local environment, its electronic configuration and polarization, the probe’s lattice sites, point defects and its recombination dynamics. In the case of e-$\\gamma$ PAC, the electron mobility on the host can be further studied, e.g., as a function of temperature.

  16. Mapping vortex-like hydrodynamic flow in microfluidic networks using fluorescence correlation spectroscopy

    Ke Liu; Yu Tian; Burrows, Sean M.; Reif, Randall D. [Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409-1061 (United States); Pappas, Dimitri, E-mail: d.pappas@ttu.edu [Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409-1061 (United States)

    2009-09-28

    The ability to quickly measure flow parameters in microfluidic devices is critical for micro total analysis system ({mu}TAS) applications. Macrofluidic methods to assess flow suffer from limitations that have made conventional methods unsuitable for the flow behavior profiling. Single molecule fluorescence correlation spectroscopy (FCS) has been employed in our study to characterize the fluidic vortex generating at a T-shape junction of microscale channels. Due to its high spatial and temporal resolution, the corresponding magnitudes relative to different flow rates in the main channel can be quantitatively differentiated using flow time ({tau}{sub F}) measurements of dye molecules traversing the detection volume in buffer solution. Despite the parabolic flow in the channel upstream, a heterogeneous distribution of flow has been detected across the channel intersection. In addition, our current observations also confirmed the aspect of vortex-shaped flow in low-shear design that was developed previously for cell culture. This approach not only overcomes many technical barriers for examining hydrodynamic vortices and movements in miniature structures without physically integrating any probes, but it is also especially useful for the hydrodynamic studies in polymer-glass based micro -reactor and -mixer.

  17. Quantitative measurement of intracellular transport of nanocarriers by spatio-temporal image correlation spectroscopy

    Coppola, S; Pozzi, D; De Sanctis, S Candeloro; Caracciolo, G; Digman, M A; Gratton, E

    2013-01-01

    Spatio-temporal image correlation spectroscopy (STICS) is a powerful technique for assessing the nature of particle motion in complex systems although it has been rarely used to investigate the intracellular dynamics of nanocarriers so far. Here we introduce a method for characterizing the mode of motion of nanocarriers and for quantifying their transport parameters on different length scales from single-cell to subcellular level. Using this strategy we were able to study the mechanisms responsible for the intracellular transport of DOTAP–DOPC/DNA (DOTAP: 1,2-dioleoyl-3-trimethylammonium-propane; DOPC: dioleoylphosphocholine) and DC-Chol–DOPE/DNA (DC-Chol: 3β-[N-(N,N-dimethylaminoethane)-carbamoyl] cholesterol; DOPE: dioleoylphosphatidylethanolamine) lipoplexes in CHO-K1 (CHO: Chinese hamster ovary) live cells. Measurement of both diffusion coefficients and velocity vectors (magnitude and direction) averaged over regions of the cell revealed the presence of distinct modes of motion. Lipoplexes diffused slowly on the cell surface (diffusion coefficient: D ≈ 0.003 μm 2 s −1 ). In the cytosol, the lipoplexes’ motion was characterized by active transport with average velocity v ≈ 0.03 μm 2 s −1 and random motion. The method permitted us to generate an intracellular transport map showing several regions of concerted motion of lipoplexes. (paper)

  18. Longitudinal, transcranial measurement of functional activation in the rat brain by diffuse correlation spectroscopy.

    Blanco, Igor; Zirak, Peyman; Dragojević, Tanja; Castellvi, Clara; Durduran, Turgut; Justicia, Carles

    2017-10-01

    Neural activity is an important biomarker for the presence of neurodegenerative diseases, cerebrovascular alterations, and brain trauma; furthermore, it is a surrogate marker for treatment effects. These pathologies may occur and evolve in a long time-period, thus, noninvasive, transcutaneous techniques are necessary to allow a longitudinal follow-up. In the present work, we have customized noninvasive, transcutaneous, diffuse correlation spectroscopy (DCS) to localize changes in cerebral blood flow (CBF) induced by neural activity. We were able to detect changes in CBF in the somatosensory cortex by using a model of electrical forepaw stimulation in rats. The suitability of DCS measurements for longitudinal monitoring was demonstrated by performing multiple sessions with the same animals at different ages (from 6 to 18 months). In addition, functional DCS has been cross-validated by comparison with functional magnetic resonance imaging (fMRI) in the same animals in a subset of the time-points. The overall results obtained with transcutaneous DCS demonstrates that it can be utilized in longitudinal studies safely and reproducibly to locate changes in CBF induced by neural activity in the small animal brain.

  19. Quantitative measurement of intracellular transport of nanocarriers by spatio-temporal image correlation spectroscopy

    Coppola, S.; Pozzi, D.; Candeloro De Sanctis, S.; Digman, M. A.; Gratton, E.; Caracciolo, G.

    2013-03-01

    Spatio-temporal image correlation spectroscopy (STICS) is a powerful technique for assessing the nature of particle motion in complex systems although it has been rarely used to investigate the intracellular dynamics of nanocarriers so far. Here we introduce a method for characterizing the mode of motion of nanocarriers and for quantifying their transport parameters on different length scales from single-cell to subcellular level. Using this strategy we were able to study the mechanisms responsible for the intracellular transport of DOTAP-DOPC/DNA (DOTAP: 1,2-dioleoyl-3-trimethylammonium-propane; DOPC: dioleoylphosphocholine) and DC-Chol-DOPE/DNA (DC-Chol: 3β-[N-(N,N-dimethylaminoethane)-carbamoyl] cholesterol; DOPE: dioleoylphosphatidylethanolamine) lipoplexes in CHO-K1 (CHO: Chinese hamster ovary) live cells. Measurement of both diffusion coefficients and velocity vectors (magnitude and direction) averaged over regions of the cell revealed the presence of distinct modes of motion. Lipoplexes diffused slowly on the cell surface (diffusion coefficient: D ≈ 0.003 μm2 s-1). In the cytosol, the lipoplexes’ motion was characterized by active transport with average velocity v ≈ 0.03 μm2 s-1 and random motion. The method permitted us to generate an intracellular transport map showing several regions of concerted motion of lipoplexes.

  20. Electrohydrodynamic properties of succinoglycan as probed by fluorescence correlation spectroscopy, potentiometric titration and capillary electrophoresis.

    Duval, Jérôme F L; Slaveykova, Vera I; Hosse, Monika; Buffle, Jacques; Wilkinson, Kevin J

    2006-10-01

    The electrostatic, hydrodynamic and conformational properties of aqueous solutions of succinoglycan have been analyzed by fluorescence correlation spectroscopy (FCS), proton titration, and capillary electrophoresis (CE) over a large range of pH values and electrolyte (NaCl) concentrations. Using the theoretical formalism developed previously for the electrokinetic properties of soft, permeable particles, a quantitative analysis for the electro-hydrodynamics of succinoglycan is performed by taking into account, in a self-consistent manner, the measured values of the diffusion coefficients, electric charge densities, and electrophoretic mobilities. For that purpose, two limiting conformations for the polysaccharide in solution are tested, i.e. succinoglycan behaves as (i) a spherical, random coil polymer or (ii) a rodlike particle with charged lateral chains. The results show that satisfactory modeling of the titration data for ionic strengths larger than 50 mM can be accomplished using both geometries over the entire range of pH values. Electrophoretic mobilities measured for sufficiently large pH values (pH > 5-6) are in line with predictions based on either model. The best manner to discriminate between these two conceptual models is briefly discussed. For low pH values (pH < 5), both models indicate aggregation, resulting in an increase of the hydrodynamic permeability and a decrease of the diffusion coefficient.

  1. Heat shock-induced interactions among nuclear HSFs detected by fluorescence cross-correlation spectroscopy

    Pack, Chan-Gi, E-mail: changipack@amc.seoul.kr [Asan Institute for Life Sciences, University of Ulsan, College of Medicine, Asan Medical Center, Seoul 138-736 (Korea, Republic of); Ahn, Sang-Gun [Dept. of Pathology, College of Dentistry, Chosun University, Seosuk-dong, Dong-gu, Gwangju 501-759 (Korea, Republic of)

    2015-07-31

    The cellular response to stress is primarily controlled in cells via transcriptional activation by heat shock factor 1 (HSF1). HSF1 is well-known to form homotrimers for activation upon heat shock and subsequently bind to target DNAs, such as heat-shock elements, by forming stress granules. A previous study demonstrated that nuclear HSF1 and HSF2 molecules in live cells interacted with target DNAs on the stress granules. However, the process underlying the binding interactions of HSF family in cells upon heat shock remains unclear. This study demonstrate for the first time that the interaction kinetics among nuclear HSF1, HSF2, and HSF4 upon heat shock can be detected directly in live cells using dual color fluorescence cross-correlation spectroscopy (FCCS). FCCS analyses indicated that the binding between HSFs was dramatically changed by heat shock. Interestingly, the recovery kinetics of interaction between HSF1 molecules after heat shock could be represented by changes in the relative interaction amplitude and mobility. - Highlights: • The binding interactions among nuclear HSFs were successfully detected. • The binding kinetics between HSF1s during recovery was quantified. • HSF2 and HSF4 strongly formed hetero-complex, even before heat shock. • Nuclear HSF2 and HSF4 bound to HSF1 only after heat shock.

  2. Exchange-dynamics of a neutral hydrophobic dye in micellar solutions studied by Fluorescence Correlation Spectroscopy.

    Bordello, Jorge; Novo, Mercedes; Al-Soufi, Wajih

    2010-05-15

    The dynamics of the exchange of the moderately hydrophobic neutral dye Coumarine 152 between the aqueous phase and the phase formed by neutral Triton X-100 micelles is studied by Fluorescence Correlation Spectroscopy. The changes in the photophysical properties of the dye in presence of the micelles are discussed. The low quantum yield, the low saturation threshold and the necessary high energetic excitation of this dye requires a careful selection of the experimental conditions in order to obtain dynamic and diffusional properties with reasonable precision. It is shown that the contrast between the brightness of free and bound dye has a strong influence on the sensitivity of the FCS experiment. The entry rate constant of the dye to the micelles, k(+)=(0.8±0.3)×10(10) M(-1) s(-1), is very near to the diffusion controlled limit. The high association equilibrium constant of K=(129±3)×10(3) M(-1) is mainly determined by the low exit rate constant, k(-)=(0.6±0.2)×10(5) s(-1). Copyright © 2010 Elsevier Inc. All rights reserved.

  3. Decreased microvascular cerebral blood flow assessed by diffuse correlation spectroscopy after repetitive concussions in mice.

    Buckley, Erin M; Miller, Benjamin F; Golinski, Julianne M; Sadeghian, Homa; McAllister, Lauren M; Vangel, Mark; Ayata, Cenk; Meehan, William P; Franceschini, Maria Angela; Whalen, Michael J

    2015-12-01

    Repetitive concussions are associated with long-term cognitive dysfunction that can be attenuated by increasing the time intervals between concussions; however, biomarkers of the safest rest interval between injuries remain undefined. We hypothesize that deranged cerebral blood flow (CBF) is a candidate biomarker for vulnerability to repetitive concussions. Using a mouse model of human concussion, we examined the effect of single and repetitive concussions on cognition and on an index of CBF (CBFi) measured with diffuse correlation spectroscopy. After a single mild concussion, CBFi was reduced by 35±4% at 4 hours (Pconcussions spaced 1 day apart, CBFi was also reduced from preinjury levels 4 hours after each concussion but had returned to preinjury levels by 72 hours after the final concussion. Interestingly, in this repetitive concussion model, lower CBFi values measured both preinjury and 4 hours after the third concussion were associated with worse performance on the Morris water maze assessed 72 hours after the final concussion. We conclude that low CBFi measured either before or early on in the evolution of injury caused by repetitive concussions could be a useful predictor of cognitive outcome.

  4. Fluorescence correlation spectroscopy to study antibody binding and stoichiometry of complexes

    Swift, Kerry M.; Matayoshi, Edmund D.

    2008-02-01

    FCS (fluorescence correlation spectroscopy) was used to study the association at the single molecule level of tumor necrosis factor alpha (TNF-α) and two of its protein antagonists Humira (TM) (adalimumab), a fully humanized monoclonal antibody, and Enbrel (TM) (etanercept), a soluble form of the TNF receptor. Single molecule approaches potentially have the advantage not only of enhanced sensitivity, but also of observing at equilibrium the details that would otherwise be lost in classical ensemble experiments where heterogeneity is averaged. We prepared fluorescent conjugates of the protein drugs and their biological target, the trimeric soluble form of TNF-α. The bivalency of adalimumab and the trimeric nature of TNF-α potentially allow several forms of associative complexes that may differ in stoichiometry. Detailed knowledge of this reaction may be relevant to understanding adalimumab's pharmacological properties. Our FCS data showed that a single trimeric TNF-α can bind up to three adalimumab molecules. Under some conditions even larger complexes are formed, apparently the result of cross-linking of TNF-α trimers by adalimumab. In addition, distinct differences between Humira and Enbrel were observed in their association with TNF-α.

  5. Dynamics and rheology under continuous shear flow studied by x-ray photon correlation spectroscopy

    Fluerasu, Andrei [Brookhaven National Laboratory, NSLS-II, Upton, NY 11973 (United States); Kwasniewski, Pawel; Caronna, Chiara; Madsen, Anders [European Synchrotron Radiation Facility, ID10 (Troika), Grenoble 38043 (France); Destremaut, Fanny; Salmon, Jean-Baptiste [LOF, UMR 5258 CNRS-Rhodia Bordeaux 1, 33608 Pessac (France)], E-mail: fluerasu@bnl.gov

    2010-03-15

    X-ray photon correlation spectroscopy (XPCS) has emerged as a unique technique allowing the measurement of dynamics of materials on mesoscopic lengthscales. One of the most common problems associated with the use of bright x-ray beams is beam-induced radiation damage, and this is likely to become an even more limiting factor at future synchrotron and free-electron laser sources. Flowing the sample during data acquisition is one of the simplest methods allowing the radiation damage to be limited. In addition to distributing the dose over many different scatterers, the method also enables new functionalities such as time-resolved studies. Here, we further develop a recently proposed experimental technique that combines XPCS and continuously flowing samples. More specifically, we use a model colloidal suspension to show how the macroscopic advective response to flow and the microscopic dissipative dynamics (diffusion) can be quantified from the x-ray data. Our results show very good quantitative agreement with a Poisseuille-flow hydrodynamical model combined with Brownian mechanics. The method has many potential applications, e.g. in the study of dynamics of glasses and gels under continuous shear/flow, protein aggregation processes and the interplay between dynamics and rheology in complex fluids.

  6. Benchmarking fully analytic DFT force fields for vibrational spectroscopy: A study on halogenated compounds

    Pietropolli Charmet, Andrea; Cornaton, Yann

    2018-05-01

    This work presents an investigation of the theoretical predictions yielded by anharmonic force fields having the cubic and quartic force constants are computed analytically by means of density functional theory (DFT) using the recursive scheme developed by M. Ringholm et al. (J. Comput. Chem. 35 (2014) 622). Different functionals (namely B3LYP, PBE, PBE0 and PW86x) and basis sets were used for calculating the anharmonic vibrational spectra of two halomethanes. The benchmark analysis carried out demonstrates the reliability and overall good performances offered by hybrid approaches, where the harmonic data obtained at the coupled cluster with single and double excitations level of theory augmented by a perturbational estimate of the effects of connected triple excitations, CCSD(T), are combined with the fully analytic higher order force constants yielded by DFT functionals. These methods lead to reliable and computationally affordable calculations of anharmonic vibrational spectra with an accuracy comparable to that yielded by hybrid force fields having the anharmonic force fields computed at second order Møller-Plesset perturbation theory (MP2) level of theory using numerical differentiation but without the corresponding potential issues related to computational costs and numerical errors.

  7. Hypoxia-ischemic encephalopathy in full-term neonate: correlation proton MR spectroscopy with MR imaging

    Fan Guoguang; Wu Zhenhua; Chen Liying; Guo Qiyong; Ye Binbin; Mao Jian

    2003-01-01

    Introduction: To evaluate 1 H Magnetic Resonance Spectroscopy ( 1 HMRS) in the diagnosis of hypoxia-ischemic encephalopathy (HIE) of full-term neonates correlated with Magnetic Resonance Imaging (MRI). Materials and methods: Thirty-eight cases of full-term neonates diagnosed as HIE clinically were selected to perform MRI and 1 HMRS examination. The ages ranged from 7 to 17 days, with median age of 8.2 days. In which, 26 cases were followed up and/or MRI reexamined at 6 months of age or later. Eight healthy neonates, with no evidence of birth asphyxia, also underwent 1 HMRS for comparison. SE sequences were used for routine MR examination; point resolved spectroscopy sequence was required for 1 HMRS. The metabolites in the spectra includes: N-acetylaspartate (NAA), choline compounds (CHO), creatine compounds (CR), myo-inositol (MI), lactate (LAC), glutamate and glutamine (Glu-Gln). Results: The peaks of NAA were fall in two cases; the peaks of LAC, which were elevated, appeared as typical double-peaks appearance in 26 cases; the peaks of Glu-Gln, which were also elevated, appeared as zigzag appearance in nine cases. The peaks of CR were decreased in 11 cases, while those of MI were increased in seven cases. Mild type of lesions was present on MRI in 12 cases whose LAC/CR ratio lower than 0.5; mild and moderate types of lesions were present in 15 cases whose LAC/CR ratio between 0.5 and 1.5. Whereas, nine cases of severe lesions and two cases of moderate lesions were present on MRI in 11 cases whose LAC/CR ratio greater than 1.5. Twenty-six of 38 cases were followed up and/or MRI reexamined after 6 months, in which, sequelae were present in 12 cases. Among them, eight cases of sequelae in nine cases whose LAC/CR ratio greater than 1.5 were present (account for 88.89%). Conclusion: 1 HMRS plays an important role to diagnose and predict outcome of HIE

  8. Characterization by Fourier transform infrared spectroscopy (FT-IR) and 2D IR correlation spectroscopy of a carbosilane dendrimer with peripheral ammonium groups

    Popescu, Maria-Cristina, E-mail: cpopescu@icmpp.ro [' Petru Poni' Institute of Macromolecular Chemistry (Romania); Gomez, Rafael; Mata, Fco Javier de la; Rasines, Beatriz [Universidad de Alcala, Departamento de Quimica Inorganica (Spain); Simionescu, Bogdan C. [' Petru Poni' Institute of Macromolecular Chemistry (Romania)

    2013-06-15

    Fourier transform infrared spectroscopy and 2D correlation spectroscopy were used to study the microstructural changes occurring on heating of a new carbosilane dendrimer with peripheral ammonium groups. Temperature-dependent spectral variations in the 3,010-2,710, 1,530-1,170, and 1,170-625 cm{sup -1} regions were monitored during the heating process. The dependence, on temperature, of integral absorptions and position of spectral bands was established and the spectral modifications associated with molecular conformation rearrangements, allowing molecular shape changes, were found. Before 180 Degree-Sign C, the studied carbosilane dendrimer proved to be stable, while at higher temperatures it oxidizes and Si-O groups appear. 2D IR correlation spectroscopy gives new information about the effect of temperature on the structure and dynamics of the system. Synchronous and asynchronous spectra indicate that, at low temperature, conformational changes of CH{sub 3} and CH{sub 3}-N{sup +} groups take place first. With increasing temperature, the intensity variation of the CH{sub 2}, C-N, Si-C and C-C groups from the dendritic core is faster than that of the terminal units. This indicates that, with increasing temperature, the segments of the dendritic core obtain enough energy to change their conformation more easily as compared to the terminal units, due to their internal flexibility.

  9. Characterization by Fourier transform infrared spectroscopy (FT-IR) and 2D IR correlation spectroscopy of a carbosilane dendrimer with peripheral ammonium groups

    Popescu, Maria-Cristina; Gómez, Rafael; Mata, Fco Javier de la; Rasines, Beatriz; Simionescu, Bogdan C.

    2013-01-01

    Fourier transform infrared spectroscopy and 2D correlation spectroscopy were used to study the microstructural changes occurring on heating of a new carbosilane dendrimer with peripheral ammonium groups. Temperature-dependent spectral variations in the 3,010–2,710, 1,530–1,170, and 1,170–625 cm −1 regions were monitored during the heating process. The dependence, on temperature, of integral absorptions and position of spectral bands was established and the spectral modifications associated with molecular conformation rearrangements, allowing molecular shape changes, were found. Before 180 °C, the studied carbosilane dendrimer proved to be stable, while at higher temperatures it oxidizes and Si–O groups appear. 2D IR correlation spectroscopy gives new information about the effect of temperature on the structure and dynamics of the system. Synchronous and asynchronous spectra indicate that, at low temperature, conformational changes of CH 3 and CH 3 –N + groups take place first. With increasing temperature, the intensity variation of the CH 2 , C–N, Si–C and C–C groups from the dendritic core is faster than that of the terminal units. This indicates that, with increasing temperature, the segments of the dendritic core obtain enough energy to change their conformation more easily as compared to the terminal units, due to their internal flexibility.

  10. Raman spectroscopy compared against traditional predictors of shear force in lamb m. longissimus lumborum.

    Fowler, Stephanie M; Schmidt, Heinar; van de Ven, Remy; Wynn, Peter; Hopkins, David L

    2014-12-01

    A Raman spectroscopic hand held device was used to predict shear force (SF) of 80 fresh lamb m. longissimus lumborum (LL) at 1 and 5days post mortem (PM). Traditional predictors of SF including sarcomere length (SL), particle size (PS), cooking loss (CL), percentage myofibrillar breaks and pH were also measured. SF values were regressed against Raman spectra using partial least squares regression and against the traditional predictors using linear regression. The best prediction of shear force values used spectra at 1day PM to predict shear force at 1day which gave a root mean square error of prediction (RMSEP) of 13.6 (Null=14.0) and the R(2) between observed and cross validated predicted values was 0.06 (R(2)cv). Overall, for fresh LL, the predictability SF, by either the Raman hand held probe or traditional predictors was low. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Characterization of chemically and enzymatically treated hemp fibres using atomic force microscopy and spectroscopy

    George, Michael; Mussone, Paolo G. [Biorefining Conversions and Fermentations Laboratory, Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada T6E 2P5 (Canada); Abboud, Zeinab [Biorefining Conversions and Fermentations Laboratory, Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada T6E 2P5 (Canada); Department of Physics, University of Guelph, Guelph, ON, Canada N1G 2W1 (Canada); Bressler, David C., E-mail: david.bressler@ualberta.ca [Biorefining Conversions and Fermentations Laboratory, Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, Canada T6E 2P5 (Canada)

    2014-09-30

    The mechanical and moisture resistance properties of natural fibre reinforced composites are dependent on the adhesion between the matrix of choice and the fibre. The main goal of this study was to investigate the effect of NaOH swelling of hemp fibres prior to enzymatic treatment and a novel chemical sulfonic acid method on the physical properties of hemp fibres. The colloidal properties of treated hemp fibres were studied exclusively using an atomic force microscope. AFM imaging in tapping mode revealed that each treatment rendered the surface topography of the hemp fibres clean and exposed the individual fibre bundles. Hemp fibres treated with laccase had no effect on the surface adhesion forces measured. Interestingly, mercerization prior to xylanase + cellulase and laccase treatments resulted in greater enzyme access evident in the increased adhesion force measurements. Hemp fibres treated with sulfonic acid showed an increase in surface de-fibrillation and smoothness. A decrease in adhesion forces for 4-aminotoulene-3-sulfonic acid (AT3S) treated fibres suggested a reduction in surface polarity. This work demonstrated that AFM can be used as a tool to estimate the surface forces and roughness for modified fibres and that enzymatic coupled with chemical methods can be used to improve the surface properties of natural fibres for composite applications. Further, this work is one of the first that offers some insight into the effect of mercerization prior to enzymes and the effect on the surface topography. AFM will be used to selectively screen treated fibres for composite applications based on the adhesion forces associated with the colloidal interface between the AFM tip and the fibre surfaces.

  12. Repurposing a Benchtop Centrifuge for High-Throughput Single-Molecule Force Spectroscopy.

    Yang, Darren; Wong, Wesley P

    2018-01-01

    We present high-throughput single-molecule manipulation using a benchtop centrifuge, overcoming limitations common in other single-molecule approaches such as high cost, low throughput, technical difficulty, and strict infrastructure requirements. An inexpensive and compact Centrifuge Force Microscope (CFM) adapted to a commercial centrifuge enables use by nonspecialists, and integration with DNA nanoswitches facilitates both reliable measurements and repeated molecular interrogation. Here, we provide detailed protocols for constructing the CFM, creating DNA nanoswitch samples, and carrying out single-molecule force measurements.

  13. Dynamics of shearing force and its correlations with chemical compositions and dry matter digestibility of stylo ( stem

    Xuejuan Zi

    2017-12-01

    Full Text Available Objective The study explored the dynamics of shearing force and its correlation with chemical compositions and in vitro dry matter digestibility (IVDMD of stylo. Methods The shearing force, diameter, linear density, chemical composition, and IVDMD of different height stylo stem were investigated. Linear regression analysis was done to determine the relationships between the shearing force and cut height, diameter, chemical composition, or IVDMD. Results The results showed that shearing force of stylo stem increased with plant height increasing and the crude protein (CP content and IVDMD decreased but fiber content increased over time, resulting in decreased forage value. In addition, tall stem had greater shearing force than short stem. Moreover, shearing force is positively correlated with stem diameter, linear density and fiber fraction, but negatively correlated with CP content and IVDMD. Conclusion Overall, shearing force is an indicator more direct, easier and faster to measure than chemical composition and digestibility for evaluation of forage nutritive value related to animal performance. Therefore, it can be used to evaluate the nutritive value of stylo.

  14. Force Spectroscopy of Hyaluronan by AFM; From H-bonded Networks Towards Single Chain Behavior

    Giannotti, M.I.; Rinaudo, Marguerite; Vancso, Gyula J.

    2007-01-01

    The conformational behavior of hyaluronan (HA) polysaccharide chains in aqueous NaCl solution was characterized directly at the single-molecule level. This comunication reports on one of the first single-chain atomic force microscopy (AFM) experiments performed at variable temperatures,

  15. Quantitative analysis of tip-sample interaction in non-contact scanning force spectroscopy

    Palacios-Lidon, Elisa; Colchero, Jaime

    2006-01-01

    Quantitative characterization of tip-sample interaction in scanning force microscopy is fundamental for optimum image acquisition as well as data interpretation. In this work we discuss how to characterize the electrostatic and van der Waals contribution to tip-sample interaction in non-contact scanning force microscopy precisely. The spectroscopic technique presented is based on the simultaneous measurement of cantilever deflection, oscillation amplitude and frequency shift as a function of tip-sample voltage and tip-sample distance as well as on advanced data processing. Data are acquired at a fixed lateral position as interaction images, with the bias voltage as fast scan, and tip-sample distance as slow scan. Due to the quadratic dependence of the electrostatic interaction with tip-sample voltage the van der Waals force can be separated from the electrostatic force. Using appropriate data processing, the van der Waals interaction, the capacitance and the contact potential can be determined as a function of tip-sample distance. The measurement of resonance frequency shift yields very high signal to noise ratio and the absolute calibration of the measured quantities, while the acquisition of cantilever deflection allows the determination of the tip-sample distance

  16. Force Spectroscopy of Collagen Fibers to Investigate Their Mechanical Properties and Structural Organization

    Gutsmann, Thomas; Fantner, Georg E.; Kindt, Johannes H.; Venturoni, Manuela; Danielsen, Signe; Hansma, Paul K.

    2004-01-01

    Tendons are composed of collagen and other molecules in a highly organized hierarchical assembly, leading to extraordinary mechanical properties. To probe the cross-links on the lower level of organization, we used a cantilever to pull substructures out of the assembly. Advanced force probe technology, using small cantilevers (length

  17. Atomic force microscope with combined FTIR-Raman spectroscopy having a micro thermal analyzer

    Fink, Samuel D [Aiken, SC; Fondeur, Fernando F [North Augusta, SC

    2011-10-18

    An atomic force microscope is provided that includes a micro thermal analyzer with a tip. The micro thermal analyzer is configured for obtaining topographical data from a sample. A raman spectrometer is included and is configured for use in obtaining chemical data from the sample.

  18. Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains

    Šachl, Radek; Hof, Martin; Bergstrand, Jan; Widengren, Jerker

    2016-01-01

    It has been shown by means of simulations that spot variation fluorescence correlation spectroscopy (sv-FCS) can be used for the identification and, to some extent, also characterization of immobile lipid nanodomains in model as well as cellular plasma membranes. However, in these simulations, the nanodomains were assumed to be stationary, whereas they actually tend to move like the surrounding lipids. In the present study, we investigated how such domain movement influences the diffusion time/spot-size dependence observed in FCS experiments, usually referred to as ‘diffusion law’ analysis. We show that domain movement might mask the effects of the ‘anomalous’ diffusion characteristics of membrane lipids or proteins predicted for stationary domains, making it difficult to identify such moving nanodomains by sv-FCS. More specifically, our simulations indicate that (i) for domains moving up to a factor of 2.25 slower than the surrounding lipids, such impeded diffusion cannot be observed and the diffusion behaviour of the proteins or lipids is indistinguishable from that of freely diffusing molecules, i.e. nanodomains are not detected; (ii) impeded protein/lipid diffusion behaviour can be observed in experiments where the radii of the detection volume are similar in size to the domain radii, the domain diffusion is about 10 times slower than that of the lipids, and the probes show a high affinity to the domains; and (iii) presence of nanodomains can only be reliably detected by diffraction limited sv-FCS when the domains move very slowly (about 200 times slower than the lipid diffusion). As nanodomains are expected to be in the range of tens of nanometres and most probes show low affinities to such domains, sv-FCS is limited to stationary domains and/or STED-FCS. However, even for that latter technique, diffusing domains smaller than 50 nm in radius are hardly detectable by FCS diffusion time/spot-size dependencies. (paper)

  19. Development of an X-ray delay unit for correlation spectroscopy and pump-probe experiments

    Roseker, Wojciech

    2008-07-15

    Probing condensed matter on time scales ranging from femtoseconds to nanoseconds will be one of the key topics for future X-ray Free Electron Laser (XFEL) sources. The accessible time windows are, however, compromised by the intrinsic time structure of the sources. One way to overcome this limitation is the usage of a time delay unit. A prototype device capable of splitting an X-ray pulse into two adjustable fractions, delaying one of them with the aim to perform X-ray Photon Correlation Spectroscopy and pump-probe type studies was designed and manufactured. The device utilizes eight perfect crystals in vertical 90 scattering geometry. Its performance has been verified with 8.39 keV and 12.4 keV Xrays at various synchrotron sources. The measured throughput of the device with a Si(333) monochromator at 8.39 keV under ambient conditions is 0.6%. The stability was verified at 12.4 keV and operation without realignment and feedback was possible for more than 30 minutes. Time delays up to 2.95 ns have been achieved. The highest resolution achieved in an experiment was 15.4 ps, a value entirely determined by the diagnostics system. The influence of the delay unit optics on the coherence properties of the beam was investigated by means of Fraunhofer diffraction and static speckle analysis. The obtained high fringe visibility and contrast values larger than 23% indicate the feasibility of performing coherence based experiments with the delay line. (orig.)

  20. Indirectly detected chemical shift correlation NMR spectroscopy in solids under fast magic angle spinning

    Mao, Kanmi [Iowa State Univ., Ames, IA (United States)

    2011-01-01

    The development of fast magic angle spinning (MAS) opened up an opportunity for the indirect detection of insensitive low-γ nuclei (e.g., 13C and 15N) via the sensitive high-{gamma} nuclei (e.g., 1H and 19F) in solid-state NMR, with advanced sensitivity and resolution. In this thesis, new methodology utilizing fast MAS is presented, including through-bond indirectly detected heteronuclear correlation (HETCOR) spectroscopy, which is assisted by multiple RF pulse sequences for 1H-1H homonuclear decoupling. Also presented is a simple new strategy for optimization of 1H-1H homonuclear decoupling. As applications, various classes of materials, such as catalytic nanoscale materials, biomolecules, and organic complexes, are studied by combining indirect detection and other one-dimensional (1D) and two-dimensional (2D) NMR techniques. Indirectly detected through-bond HETCOR spectroscopy utilizing refocused INEPT (INEPTR) mixing was developed under fast MAS (Chapter 2). The time performance of this approach in 1H detected 2D 1H{l_brace}13C{r_brace} spectra was significantly improved, by a factor of almost 10, compared to the traditional 13C detected experiments, as demonstrated by measuring naturally abundant organic-inorganic mesoporous hybrid materials. The through-bond scheme was demonstrated as a new analytical tool, which provides complementary structural information in solid-state systems in addition to through-space correlation. To further benefit the sensitivity of the INEPT transfer in rigid solids, the combined rotation and multiple-pulse spectroscopy (CRAMPS) was implemented for homonuclear 1H decoupling under fast MAS (Chapter 3). Several decoupling schemes (PMLG5m$\\bar{x}$, PMLG5mm$\\bar{x}$x and SAM3) were analyzed to maximize the performance of through-bond transfer based

  1. Correlation effects in magnetic materials: An ab initio investigation on electronic structure and spectroscopy

    Minár, J.; Braun, J.; Ebert, H.

    2013-01-01

    Highlights: ► We compare spin-resolved ARPES data of ferromagnetic 3d transition metals to many-body LSDA + DMFT based spectroscopic calculations. ► We document LSDA + DMFT provides a detailed and reliable interpretation of the data. ► We demonstrate that local correlations are dominant in Ni, whereas non-local correlations are important in Fe and Co. ► We reproduce the 6 eV satellite structure in ferromagnetic Ni LDSDA + DMFT in combination with the one-step model of photoemission provides a more or less complete description of the electronic structure of Fe, Co and Ni. -- Abstract: Various technical developments enlarged the potential of angle-resolved photoemission spectroscopy (ARPES) tremendously during the last two decades. In particular improved momentum and energy resolution in combination with spin-resolution as well as the use of photon energies from few eV up to several keV makes ARPES a rather unique tool to investigate the electronic properties of solids and surfaces. Obviously, this rises the need for a corresponding theoretical formalism that allows to accompany experimental ARPES studies in an adequate way. As will be demonstrated by several examples this goal could be achieved by various recent developments on the basis of density functional theory (DFT) in combination with dynamical mean field theory (DMFT) and with the one-step model of photoemission (1SM). A concrete realization of electronic structure calculations in the framework of multiple scattering theory further more provides direct access to the spectral function of the initial states via the one-electron Green function. Based on this bare spectral function matrix-element and final-state effects as well as surface related features may be calculated in addition using the one-step formalism that offers the possibility to analyse corresponding angle-resolved photoemission experiments in a quantitative sense. The impact of chemical disorder can be handled by means of the coherent

  2. Verification of Ganoderma (lingzhi) commercial products by Fourier Transform infrared spectroscopy and two-dimensional IR correlation spectroscopy

    Choong, Yew-Keong; Sun, Su-Qin; Zhou, Qun; Lan, Jin; Lee, Han-Lim; Chen, Xiang-Dong

    2014-07-01

    Ganoderma commercial products are typically based on two sources, raw material (powder form and/or spores) and extract (water and/or solvent). This study compared three types of Ganoderma commercial products using 1 Dimensional Fourier Transform infrared and second derivative spectroscopy. The analyzed spectra of Ganoderma raw material products were compared with spectra of cultivated Ganoderma raw material powder from different mushroom farms in Malaysia. The Ganoderma extract product was also compared with three types of cultivated Ganoderma extracts. Other medicinal Ganoderma contents in commercial extract product that included glucan and triterpenoid were analyzed by using FTIR and 2DIR. The results showed that water extract of cultivated Ganoderma possessed comparable spectra with that of Ganoderma product water extract. By comparing the content of Ganoderma commercial products using FTIR and 2DIR, product content profiles could be detected. In addition, the geographical origin of the Ganoderma products could be verified by comparing their spectra with Ganoderma products from known areas. This study demonstrated the possibility of developing verification tool to validate the purity of commercial medicinal herbal and mushroom products.

  3. Characterization of vacancy type defects in Electronic Materials by Positron Lifetime and Age-Momentum Correlation Spectroscopy

    Suzuki, Ryoichi; Ohdaira, Toshiyuki

    2002-03-01

    Positron annihilation spectroscopy is known to be sensitive to vacancy type defects. At the National Institute of Advanced Industrial Science and Technology (AIST) Japan, the authors have developed a measurement system which enables us to perform depth-selective positron annihilation lifetime spectroscopy (PALS) and positron age-momentum correlation (AMOC) spectroscopy with an intense slow positron beam. PALS gives us information on the size of vacancies whereas AMOC gives us information on not only vacancy sizes but also impurities or chemical environments. Using this system, we have carried out defect characterization experiments on various electronic materials, e.g. ion implanted Si, SiO2/Si, MOS, CVD or SOD (spin-on-dielectric) grown low dielectric insulator films, etc.

  4. Nanoporous Structure of Bone Matrix at Osteoporosis from Data of Atomic Force Microscopy and IR Spectroscopy

    A. A. Gaidash

    2011-01-01

    Full Text Available It was found that in an osteoporotic bone the fraction of nanosized pores decreases, the mineral phase amorphizes, hydrated shells around mineralized particles of the bone matrix thicken, and adhesion forces increase. This contributes to the formation of water clusters similar to bulk water clusters compared to the healthy bone tissue and leads to the accumulation of more viscous liquid with increased intermolecular interaction forces in the pores of the bone matrix. Given this, the rates of chemical reactions proceeding in the water phase of ultrathin channels of general parts of collagen fibrils decrease. Ultimately, nanopores of collagen-apatite interfaces lose, to a certain extent, the capability of catalyzing the hydroxyapatite crystallization.

  5. Correlations between forced oscillation technique parameters and pulmonary densitovolumetry values in patients with acromegaly

    G.B. Camilo

    2015-01-01

    Full Text Available The aims of this study were to evaluate the forced oscillation technique (FOT and pulmonary densitovolumetry in acromegalic patients and to examine the correlations between these findings. In this cross-sectional study, 29 non-smoking acromegalic patients and 17 paired controls were subjected to the FOT and quantification of lung volume using multidetector computed tomography (Q-MDCT. Compared with the controls, the acromegalic patients had a higher value for resonance frequency [15.3 (10.9-19.7 vs 11.4 (9.05-17.6 Hz, P=0.023] and a lower value for mean reactance [0.32 (0.21-0.64 vs 0.49 (0.34-0.96 cm H2O/L/s2, P=0.005]. In inspiratory Q-MDCT, the acromegalic patients had higher percentages of total lung volume (TLV for nonaerated and poorly aerated areas [0.42% (0.30-0.51% vs 0.25% (0.20-0.32%, P=0.039 and 3.25% (2.48-3.46% vs 1.70% (1.45-2.15%, P=0.001, respectively]. Furthermore, the acromegalic patients had higher values for total lung mass in both inspiratory and expiratory Q-MDCT [821 (635-923 vs 696 (599-769 g, P=0.021 and 844 (650-945 vs 637 (536-736 g, P=0.009, respectively]. In inspiratory Q-MDCT, TLV showed significant correlations with all FOT parameters. The TLV of hyperaerated areas showed significant correlations with intercept resistance (rs=−0.602, P<0.001 and mean resistance (rs=−0.580, P<0.001. These data showed that acromegalic patients have increased amounts of lung tissue as well as nonaerated and poorly aerated areas. Functionally, there was a loss of homogeneity of the respiratory system. Moreover, there were correlations between the structural and functional findings of the respiratory system, consistent with the pathophysiology of the disease.

  6. Measuring protein isoelectric points by AFM-based force spectroscopy using trace amounts of sample

    Guo, Shifeng; Zhu, Xiaoying; Jańczewski, Dominik; Lee, Serina Siew Chen; He, Tao; Teo, Serena Lay Ming; Vancso, G. Julius

    2016-09-01

    Protein charge at various pH and isoelectric point (pI) values is important in understanding protein function. However, often only trace amounts of unknown proteins are available and pI measurements cannot be obtained using conventional methods. Here, we show a method based on the atomic force microscope (AFM) to determine pI using minute quantities of proteins. The protein of interest is immobilized on AFM colloidal probes and the adhesion force of the protein is measured against a positively and a negatively charged substrate made by layer-by-layer deposition of polyelectrolytes. From the AFM force-distance curves, pI values with an estimated accuracy of ±0.25 were obtained for bovine serum albumin, myoglobin, fibrinogen and ribonuclease A over a range of 4.7-9.8. Using this method, we show that the pI of the ‘footprint’ of the temporary adhesive proteins secreted by the barnacle cyprid larvae of Amphibalanus amphitrite is in the range 9.6-9.7.

  7. Correlation between in vitro Elastomeric Force Degradation and Glass Transition Temperature determined by Dynamic Mechanical Analysis

    Manu Krishnan

    2012-01-01

    Conclusion: Tg can be considered as a dependable parameter in envisaging the force properties of an elastomeric chain. Clinically, it suggested that a polymer with higher Tg can give higher or optimum levels of force for tooth movement.

  8. Detection of structurally similar adulterants in botanical dietary supplements by thin-layer chromatography and surface enhanced Raman spectroscopy combined with two-dimensional correlation spectroscopy.

    Li, Hao; Zhu, Qing xia; Chwee, Tsz sian; Wu, Lin; Chai, Yi feng; Lu, Feng; Yuan, Yong fang

    2015-07-09

    Thin-layer chromatography (TLC) coupled with surface enhanced Raman spectroscopy (SERS) has been widely used for the study of various complex systems, especially for the detection of adulterants in botanical dietary supplements (BDS). However, this method is not sufficient to distinguish structurally similar adulterants in BDS since the analogs have highly similar chromatographic and/or spectroscopic behaviors. Taking into account the fact that higher cost and more time will be required for comprehensive chromatographic separation, more efforts with respect to spectroscopy are now focused on analyzing the overlapped SERS peaks. In this paper, the combination of a TLC-SERS method with two-dimensional correlation spectroscopy (2DCOS), with duration of exposure to laser as the perturbation, is applied to solve this problem. Besides the usual advantages of the TLC-SERS method, such as its simplicity, rapidness, and sensitivity, more advantages are presented here, such as enhanced selectivity and good reproducibility, which are obtained by 2DCOS. Two chemicals with similar structures are successfully differentiated from the complex BDS matrices. The study provides a more accurate qualitative screening method for detection of BDS with adulterants, and offers a new universal approach for the analysis of highly overlapped SERS peaks. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Operational Stress and Correlates of Mental Health Among Joint Task Force Guantanamo Bay Military Personnel.

    Webb-Murphy, Jennifer A; De La Rosa, Gabriel M; Schmitz, Kimberly J; Vishnyak, Elizabeth J; Raducha, Stephanie C; Roesch, Scott C; Johnston, Scott L

    2015-12-01

    Military personnel deployed to Joint Task Force Guantanamo Bay (JTF-GTMO) faced numerous occupational stressors. As part of a program evaluation, personnel working at JTF-GTMO completed several validated self-report measures. Personnel were at the beginning, middle, or end of their deployment phase. This study presents data regarding symptoms of posttraumatic stress disorder, alcohol abuse, depression, and resilience among 498 U.S. military personnel deployed to JTF-GTMO in 2009. We also investigated individual and organizational correlates of mental health among these personnel. Findings indicated that tenure at JTF-GTMO was positively related to adverse mental health outcomes. Regression models including these variables had R2 values ranging from .02 to .11. Occupation at JTF-GTMO also related to mental health such that guards reported poorer mental health than medical staff. Reluctance to seek out mental health care was also related to mental health outcomes. Those who reported being most reluctant to seek out care tended to report poorer mental health than those who were more willing to seek out care. Results suggested that the JTF-GTMO deployment was associated with significant psychological stress, and that both job-related and attitude-related variables were important to understanding mental health symptoms in this sample. Copyright © 2015 International Society for Traumatic Stress Studies.

  10. Condom Use Behaviors and Correlates of Use in the Botswana Defence Force

    Tran, Bonnie Robin; Thomas, Anne Goldzier; Ditsela, Mooketsi; Vaida, Florin; Phetogo, Robert; Kelapile, David; Chambers, Christina; Haubrich, Richard; Shaffer, Richard

    2014-01-01

    Preventing HIV infection is a priority for militaries. HIV prevention research is needed to monitor existing programs, identify areas for modification, and develop new interventions. Correct and consistent condom use is highly effective against HIV. However, use among soldiers is lower than ideal. This study describes condom use behaviors and examines correlates of use in the Botswana Defence Force (BDF). Analyses were based on 211 male personnel, aged 18–30, who completed a cross-sectional survey that collected baseline data for an intervention study. Results showed that 51% of participants reported always using condoms, 35% used condoms most times, and 14% used condoms occasionally/never. Condom use varied by partner type and was typically higher with casual partners in comparison to regular partners. After adjustment for age and marital status, factors associated with lower condom use included excessive alcohol use, perception that using condoms reduce sexual pleasure, and having a trusted partner. However, higher levels of HIV knowledge and reports of being circumcised were protective against lower condom use. HIV interventions aimed at increasing condom use in the BDF should address condom perceptions, alcohol abuse, and issues of trust. Innovative ways to increase condom use in this population should also be explored. PMID:23970609

  11. Condom use behaviours and correlates of use in the Botswana Defence Force.

    Tran, Bonnie Robin; Thomas, Anne Goldzier; Ditsela, Mooketsi; Vaida, Florin; Phetogo, Robert; Kelapile, David; Chambers, Christina; Haubrich, Richard; Shaffer, Richard

    2013-11-01

    Preventing HIV infection is a priority for militaries. HIV prevention research is needed to monitor existing programme, identify areas for modification, and develop new interventions. Correct and consistent condom use is highly effective against HIV. However, use among soldiers is lower than ideal. This study describes condom use behaviours and examines correlates of use in the Botswana Defence Force (BDF). Analyses were based on 211 male BDF personnel, aged 18-30, who completed a cross-sectional survey that collected baseline data for an intervention study. Results showed that 51% of participants reported always using condoms, 35% used condoms most times, and 14% used condoms occasionally/never. Condom use varied by partner type and was typically higher with casual partners in comparison to regular partners. After adjustment for age and marital status, factors associated with lower condom use included excessive alcohol use, perception that using condoms reduce sexual pleasure, and having a trusted partner. However, higher levels of HIV knowledge and reports of being circumcised were protective against lower condom use. HIV interventions aimed at increasing condom use in the BDF should address condom perceptions, alcohol abuse, and issues of trust. Innovative ways to increase condom use in this population should also be explored.

  12. Combined use of atomic force microscopy, X-ray photoelectron spectroscopy, and secondary ion mass spectrometry for cell surface analysis.

    Dague, Etienne; Delcorte, Arnaud; Latgé, Jean-Paul; Dufrêne, Yves F

    2008-04-01

    Understanding the surface properties of microbial cells is a major challenge of current microbiological research and a key to efficiently exploit them in biotechnology. Here, we used three advanced surface analysis techniques with different sensitivity, probing depth, and lateral resolution, that is, in situ atomic force microscopy, X-ray photoelectron spectroscopy, and secondary ion mass spectrometry, to gain insight into the surface properties of the conidia of the human fungal pathogen Aspergillus fumigatus. We show that the native ultrastructure, surface protein and polysaccharide concentrations, and amino acid composition of three mutants affected in hydrophobin production are markedly different from those of the wild-type, thereby providing novel insight into the cell wall architecture of A. fumigatus. The results demonstrate the power of using multiple complementary techniques for probing microbial cell surfaces.

  13. Cu(2) dynamical correlations in YBa2Cu3Ox studied by Moessbauer spectroscopy on substituted 170Yb3+

    Hodges, J.A.; Bonville, P.; Imbert, P.; Jehanno, G.

    1990-01-01

    Using Moessbauer spectroscopy, we follow the thermal dependence of the fluctuation rate of the Cu(2) internal (molecular) field acting on a 170 Yb 3+ probe diluted at the Y 3+ sites. We use the fluctuations of this field to track the collective fluctuations of the nearest neighbour correlated Cu(2) moments. Results are presented for oxygen levels x = 6.0 and 6.35 and for T ≤ 80K

  14. Slow Aging Dynamics and Avalanches in a Gold-Cadmium Alloy Investigated by X-Ray Photon Correlation Spectroscopy

    Mueller, L.; Waldorf, M.; Klemradt, U.; Gutt, C.; Gruebel, G.; Madsen, A.; Finlayson, T. R.

    2011-01-01

    Results of a x-ray photon correlation spectroscopy experiment on the very weakly first order martensitic transformation of a Au 50.5 Cd 49.5 single crystal are presented. Slow non-equilibrium-dynamics are observed in a narrow temperature interval in the direct vicinity of the otherwise athermal phase transformation. These dynamics are associated with the martensite-aging effect. The dynamical aging is accompanied by an avalanchelike behavior which is identified with an incubation-time phenomenon.

  15. Slow aging dynamics and avalanches in a gold-cadmium alloy investigated by x-ray photon correlation spectroscopy.

    Müller, L; Waldorf, M; Gutt, C; Grübel, G; Madsen, A; Finlayson, T R; Klemradt, U

    2011-09-02

    Results of a x-ray photon correlation spectroscopy experiment on the very weakly first order martensitic transformation of a Au50.5Cd49.5 single crystal are presented. Slow non-equilibrium-dynamics are observed in a narrow temperature interval in the direct vicinity of the otherwise athermal phase transformation. These dynamics are associated with the martensite-aging effect. The dynamical aging is accompanied by an avalanchelike behavior which is identified with an incubation-time phenomenon.

  16. Cerebral oxygenation as measured by near-infrared spectroscopy in neonatal intensive care: correlation with arterial oxygenation.

    Hunter, Carol Lu; Oei, Ju Lee; Lui, Kei; Schindler, Timothy

    2017-07-01

    To assess correlation between cerebral oxygenation (rScO 2 ), as measured by near-infrared spectroscopy (NIRS), and arterial oxygenation (PaO 2 ), as measured by arterial blood gases, in preterm neonates. Preterm neonates interpretation of NIRS values in neonatal intensive care, and further evaluation is needed to determine the applicability of NIRS to management of preterm infants. ©2017 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  17. Causal correlation of foliar biochemical concentrations with AVIRIS spectra using forced entry linear regression

    Dawson, Terence P.; Curran, Paul J.; Kupiec, John A.

    1995-01-01

    A major goal of airborne imaging spectrometry is to estimate the biochemical composition of vegetation canopies from reflectance spectra. Remotely-sensed estimates of foliar biochemical concentrations of forests would provide valuable indicators of ecosystem function at regional and eventually global scales. Empirical research has shown a relationship exists between the amount of radiation reflected from absorption features and the concentration of given biochemicals in leaves and canopies (Matson et al., 1994, Johnson et al., 1994). A technique commonly used to determine which wavelengths have the strongest correlation with the biochemical of interest is unguided (stepwise) multiple regression. Wavelengths are entered into a multivariate regression equation, in their order of importance, each contributing to the reduction of the variance in the measured biochemical concentration. A significant problem with the use of stepwise regression for determining the correlation between biochemical concentration and spectra is that of 'overfitting' as there are significantly more wavebands than biochemical measurements. This could result in the selection of wavebands which may be more accurately attributable to noise or canopy effects. In addition, there is a real problem of collinearity in that the individual biochemical concentrations may covary. A strong correlation between the reflectance at a given wavelength and the concentration of a biochemical of interest, therefore, may be due to the effect of another biochemical which is closely related. Furthermore, it is not always possible to account for potentially suitable waveband omissions in the stepwise selection procedure. This concern about the suitability of stepwise regression has been identified and acknowledged in a number of recent studies (Wessman et al., 1988, Curran, 1989, Curran et al., 1992, Peterson and Hubbard, 1992, Martine and Aber, 1994, Kupiec, 1994). These studies have pointed to the lack of a physical

  18. Chiral three-nucleon forces and the evolution of correlations along the oxygen isotopic chain

    Cipollone, A.; Barbieri, C.; Navrátil, P.

    2015-07-01

    Background: Three-nucleon forces (3NFs) have nontrivial implications on the evolution of correlations at extreme proton-neutron asymmetries. Recent ab initio calculations show that leading-order chiral interactions are crucial to obtain the correct binding energies and neutron driplines along the O, N, and F chains [A. Cipollone, C. Barbieri, and P. Navrátil, Phys. Rev. Lett. 111, 062501 (2013), 10.1103/PhysRevLett.111.062501]. Purpose: Here we discuss the impact of 3NFs along the oxygen chain for other quantities of interest, such has the spectral distribution for attachment and removal of a nucleon, spectroscopic factors, and radii. The objective is to better delineate the general effects of 3NFs on nuclear correlations. Methods: We employ self-consistent Green's function (SCGF) theory which allows a comprehensive calculation of the single-particle spectral function. For the closed subshell isotopes, 14O, 16O, 22O, 24O, and 28O, we perform calculations with the Dyson-ADC(3) method, which is fully nonperturbative and is the state of the art for both nuclear physics and quantum chemistry applications. The remaining open-shell isotopes are studied using the newly developed Gorkov-SCGF formalism up to second order. Results: We produce complete plots for the spectral distributions. The spectroscopic factors for the dominant quasiparticle peaks are found to depend very little on the leading-order (NNLO) chiral 3NFs. The latter have small impact on the calculated matter radii, which, however, are consistently obtained smaller than experiment. Similarly, single-particle spectra tend to be too spread with respect to the experiment. This effect might hinder, to some extent, the onset of correlations and screen the quenching of calculated spectroscopic factors. The most important effect of 3NFs is thus the fine tuning of the energies for the dominant quasiparticle states, which governs the shell evolution and the position of driplines. Conclusions: Although present chiral

  19. MDM2–MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance

    Moscetti I

    2016-08-01

    Full Text Available Ilaria Moscetti,1 Emanuela Teveroni,2,3 Fabiola Moretti,3 Anna Rita Bizzarri,1 Salvatore Cannistraro1 1Biophysics and Nanoscience Centre, Department DEB, Università della Tuscia, Viterbo, Italy; 2Department of Endocrinology and Metabolism, Università Cattolica di Roma, Roma, Italy; 3Institute of Cell Biology and Neurobiology, Consiglio Nazionale delle Ricerche (CNR, Roma, Italy Abstract: Murine double minute 2 (MDM2 and 4 (MDM4 are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2–MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2–MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD in the micromolar range for the MDM2–MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2–MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2–MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation. Keywords: MDM2, MDM4, atomic force spectroscopy, surface plasmon resonance

  20. Binding studies of costunolide and dehydrocostuslactone with HSA by spectroscopy and atomic force microscopy

    Gao Wenhua; Li Nana; Chen Gaopan; Xu Yanping; Chen Yaowen; Hu Shunlin; Hu Zhide

    2011-01-01

    Human serum albumin (HSA), a major plasma protein and plasma-derived therapeutic, interacts with a wide variety of drugs and native plasma metabolites. In this study the interactions of costunolide (CE) and dehydrocostuslactone (DE) with HSA were investigated by molecule modeling, atomic force microscopy (AFM), and different optical techniques. In the mechanism discussion, it was proved that fluorescence quenching of HSA by both of the drugs is a result of the formation of drug-HSA complexes. Binding parameters for the reactions were determined according to the Stern-Volmer equation and static quenching. The results of thermodynamic parameters ΔG 0 , ΔH 0 , and ΔS 0 at different temperatures indicated that hydrogen bonding interactions play a major role in the drug-HSA associations process. The binding properties were further studied by quantitative analysis of CD, FTIR, and Raman spectra. Furthermore, AFM results showed that the dimension of HSA molecules became more swollen after binding with the drugs. - Highlights: → Interactions of costunolide and dehydrocostuslactone with HSA have been investigated for the first time. → Raman spectra were used to analyze the drug-HSA interactions. → Atomic force microscopy has been used to study the topography change of HSA by addition of the drugs. → These results are important for the drugs containing costunolide and dehydrocostuslactone distribution and metabolism.

  1. Non-specific binding of Na+ and Mg2+ to RNA determined by force spectroscopy methods

    Bizarro, C. V.; Alemany, A.; Ritort, F.

    2012-01-01

    RNA duplex stability depends strongly on ionic conditions, and inside cells RNAs are exposed to both monovalent and multivalent ions. Despite recent advances, we do not have general methods to quantitatively account for the effects of monovalent and multivalent ions on RNA stability, and the thermodynamic parameters for secondary structure prediction have only been derived at 1M [Na+]. Here, by mechanically unfolding and folding a 20 bp RNA hairpin using optical tweezers, we study the RNA thermodynamics and kinetics at different monovalent and mixed monovalent/Mg2+ salt conditions. We measure the unfolding and folding rupture forces and apply Kramers theory to extract accurate information about the hairpin free energy landscape under tension at a wide range of ionic conditions. We obtain non-specific corrections for the free energy of formation of the RNA hairpin and measure how the distance of the transition state to the folded state changes with force and ionic strength. We experimentally validate the Tightly Bound Ion model and obtain values for the persistence length of ssRNA. Finally, we test the approximate rule by which the non-specific binding affinity of divalent cations at a given concentration is equivalent to that of monovalent cations taken at 100-fold concentration for small molecular constructs. PMID:22492710

  2. Microscopic theory of photon-correlation spectroscopy in strong-coupling semiconductors

    Schneebeli, Lukas

    2009-11-27

    would be a great contribution in the growing field of quantum optics in semiconductors. The efforts in QD systems are again driven by the atomic systems which not only have shown the vacuum Rabi splitting, but also the second rung, e.g. via direct spectroscopy and via photon-correlation measurements. In this thesis, it is shown that spectrally resolved photon-statistics measurements of the resonance fluorescence from realistic semiconductor quantum-dot systems allow for high contrast identification of the two-photon strong-coupling states. Using a microscopic theory, the second-rung resonance of Jaynes-Cummings ladder is analyzed and optimum excitation conditions are determined. The computed photon-statistics spectrum displays gigantic, experimentally robust resonances at the energetic positions of the second-rung emission. The resonance fluorescence equations are derived and solved for strong-coupling semiconductor quantum-dot systems using a fully quantized multimode theory and a cluster-expansion approach. A reduced model is developed to explain the origin of auto- and cross-correlation resonances in the two-photon emission spectrum of the fluorescent light. These resonances are traced back to the two-photon strong-coupling states of Jaynes-Cummings ladder. The accuracy of the reduced model is verified via numerical solution of the resonance fluorescence equations. The analysis reveals the direct relation between the squeezed-light emission and the strong-coupling states in optically excited semiconductor systems. (orig.)

  3. Discriminating Intercalative Effects of Threading Intercalator Nogalamycin, from Classical Intercalator Daunomycin, Using Single Molecule Atomic Force Spectroscopy.

    T Banerjee

    Full Text Available DNA threading intercalators are a unique class of intercalating agents, albeit little biophysical information is available on their intercalative actions. Herein, the intercalative effects of nogalamycin, which is a naturally-occurring DNA threading intercalator, have been investigated by high-resolution atomic force microscopy (AFM and spectroscopy (AFS. The results have been compared with those of the well-known chemotherapeutic drug daunomycin, which is a non-threading classical intercalator bearing structural similarity to nogalamycin. A comparative AFM assessment revealed a greater increase in DNA contour length over the entire incubation period of 48 h for nogalamycin treatment, whereas the contour length increase manifested faster in case of daunomycin. The elastic response of single DNA molecules to an externally applied force was investigated by the single molecule AFS approach. Characteristic mechanical fingerprints in the overstretching behaviour clearly distinguished the nogalamycin/daunomycin-treated dsDNA from untreated dsDNA-the former appearing less elastic than the latter, and the nogalamycin-treated DNA distinguished from the daunomycin-treated DNA-the classically intercalated dsDNA appearing the least elastic. A single molecule AFS-based discrimination of threading intercalation from the classical type is being reported for the first time.

  4. Discriminating Intercalative Effects of Threading Intercalator Nogalamycin, from Classical Intercalator Daunomycin, Using Single Molecule Atomic Force Spectroscopy.

    Banerjee, T; Banerjee, S; Sett, S; Ghosh, S; Rakshit, T; Mukhopadhyay, R

    2016-01-01

    DNA threading intercalators are a unique class of intercalating agents, albeit little biophysical information is available on their intercalative actions. Herein, the intercalative effects of nogalamycin, which is a naturally-occurring DNA threading intercalator, have been investigated by high-resolution atomic force microscopy (AFM) and spectroscopy (AFS). The results have been compared with those of the well-known chemotherapeutic drug daunomycin, which is a non-threading classical intercalator bearing structural similarity to nogalamycin. A comparative AFM assessment revealed a greater increase in DNA contour length over the entire incubation period of 48 h for nogalamycin treatment, whereas the contour length increase manifested faster in case of daunomycin. The elastic response of single DNA molecules to an externally applied force was investigated by the single molecule AFS approach. Characteristic mechanical fingerprints in the overstretching behaviour clearly distinguished the nogalamycin/daunomycin-treated dsDNA from untreated dsDNA-the former appearing less elastic than the latter, and the nogalamycin-treated DNA distinguished from the daunomycin-treated DNA-the classically intercalated dsDNA appearing the least elastic. A single molecule AFS-based discrimination of threading intercalation from the classical type is being reported for the first time.

  5. Exploring the binding of 4-thiothymidine with human serum albumin by spectroscopy, atomic force microscopy, and molecular modeling methods.

    Zhang, Juling; Gu, Huaimin; Zhang, Xiaohui

    2014-01-30

    The interaction of 4-thiothymidine (S(4)TdR) with human serum albumin (HSA) was studied by equilibrium dialysis under normal physiological conditions. In this work, the mechanism of the interaction between S(4)TdR and human serum albumin (HSA) was exploited by fluorescence, UV, CD circular, and SERS spectroscopic. Fluorescence and UV spectroscopy suggest that HSA intensities are significantly decreased when adding S(4)TdR to HAS, and the quenching mechanism of the fluorescence is static. Also, the ΔG, ΔH, and ΔS values across temperature indicated that hydrophobic interaction was the predominant binding force. The CD circular results show that there is little change in the secondary structure of HSA except the environment of amino acid changes when adding S(4)TdR to HSA. The surface-enhanced Raman scattering (SERS) shows that the interaction between S(4)TdR and HSA can be achieved through different binding sites which are probably located in the II A and III A hydrophobic pockets of HSA which correspond to Sudlow's I and II binding sites. In addition, the molecular modeling displays that S(4)TdR-HSA complex is stabilized by hydrophobic forces, which result from amino acid residues. The atomic force microscopy results revealed that the single HSA molecular dimensions were larger after interaction of 4-thiothymidine. This work would be useful to understand the state of the transportation, distribution, and metabolism of the anticancer drugs in the human body, and it could provide a useful biochemistry parameter for the development of new anti-cancer drugs and research of pharmacology mechanisms. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Force-independent distribution of correlated neural inputs to hand muscles during three-digit grasping.

    Poston, Brach; Danna-Dos Santos, Alessander; Jesunathadas, Mark; Hamm, Thomas M; Santello, Marco

    2010-08-01

    The ability to modulate digit forces during grasping relies on the coordination of multiple hand muscles. Because many muscles innervate each digit, the CNS can potentially choose from a large number of muscle coordination patterns to generate a given digit force. Studies of single-digit force production tasks have revealed that the electromyographic (EMG) activity scales uniformly across all muscles as a function of digit force. However, the extent to which this finding applies to the coordination of forces across multiple digits is unknown. We addressed this question by asking subjects (n = 8) to exert isometric forces using a three-digit grip (thumb, index, and middle fingers) that allowed for the quantification of hand muscle coordination within and across digits as a function of grasp force (5, 20, 40, 60, and 80% maximal voluntary force). We recorded EMG from 12 muscles (6 extrinsic and 6 intrinsic) of the three digits. Hand muscle coordination patterns were quantified in the amplitude and frequency domains (EMG-EMG coherence). EMG amplitude scaled uniformly across all hand muscles as a function of grasp force (muscle x force interaction: P = 0.997; cosines of angle between muscle activation pattern vector pairs: 0.897-0.997). Similarly, EMG-EMG coherence was not significantly affected by force (P = 0.324). However, coherence was stronger across extrinsic than that across intrinsic muscle pairs (P = 0.0039). These findings indicate that the distribution of neural drive to multiple hand muscles is force independent and may reflect the anatomical properties or functional roles of hand muscle groups.

  7. Explosive force production during isometric squats correlates with athletic performance in rugby union players.

    Tillin, Neale Anthony; Pain, Matthew Thomas Gerard; Folland, Jonathan

    2013-01-01

    This study investigated the association between explosive force production during isometric squats and athletic performance (sprint time and countermovement jump height). Sprint time (5 and 20 m) and jump height were recorded in 18 male elite-standard varsity rugby union players. Participants also completed a series of maximal- and explosive-isometric squats to measure maximal force and explosive force at 50-ms intervals up to 250 ms from force onset. Sprint performance was related to early phase (≤100 ms) explosive force normalised to maximal force (5 m, r = -0.63, P = 0.005; and 20 m, r = -0.54, P = 0.020), but jump height was related to later phase (>100 ms) absolute explosive force (0.51 squats (33-67%; 0.001 squats was associated with athletic performance. Specifically, sprint performance was most strongly related to the proportion of maximal force achieved in the initial phase of explosive-isometric squats, whilst jump height was most strongly related to absolute force in the later phase of the explosive-isometric squats.

  8. An integrated instrumental setup for the combination of atomic force microscopy with optical spectroscopy.

    Owen, R J; Heyes, C D; Knebel, D; Röcker, C; Nienhaus, G U

    2006-07-01

    In recent years, the study of single biomolecules using fluorescence microscopy and atomic force microscopy (AFM) techniques has resulted in a plethora of new information regarding the physics underlying these complex biological systems. It is especially advantageous to be able to measure the optical, topographical, and mechanical properties of single molecules simultaneously. Here an AFM is used that is especially designed for integration with an inverted optical microscope and that has a near-infrared light source (850 nm) to eliminate interference between the optical experiment and the AFM operation. The Tip Assisted Optics (TAO) system consists of an additional 100 x 100-microm(2) X-Y scanner for the sample, which can be independently and simultaneously used with the AFM scanner. This allows the offset to be removed between the confocal optical image obtained with the sample scanner and the simultaneously acquired AFM topography image. The tip can be positioned exactly into the optical focus while the user can still navigate within the AFM image for imaging or manipulation of the sample. Thus the tip-enhancement effect can be maximized and it becomes possible to perform single molecule manipulation experiments within the focus of a confocal optical image. Here this is applied to simultaneous measurement of single quantum dot fluorescence and topography with high spatial resolution. (c) 2006 Wiley Periodicals, Inc.

  9. Contact-resonance atomic force microscopy for nanoscale elastic property measurements: Spectroscopy and imaging

    Stan, G.; Krylyuk, S.; Davydov, A.V.; Vaudin, M.D.; Bendersky, L.A.; Cook, R.F.

    2009-01-01

    Quantitative measurements of the elastic modulus of nanosize systems and nanostructured materials are provided with great accuracy and precision by contact-resonance atomic force microscopy (CR-AFM). As an example of measuring the elastic modulus of nanosize entities, we used the CR-AFM technique to measure the out-of-plane indentation modulus of tellurium nanowires. A size-dependence of the indentation modulus was observed for the investigated tellurium nanowires with diameters in the range 20-150 nm. Over this diameter range, the elastic modulus of the outer layers of the tellurium nanowires experienced significant enhancement due to a pronounced surface stiffening effect. Quantitative estimations for the elastic moduli of the outer and inner parts of tellurium nanowires of reduced diameter are made with a core-shell structure model. Besides localized elastic modulus measurements, we have also developed a unique CR-AFM imaging capability to map the elastic modulus over a micrometer-scale area. We used this CR-AFM capability to construct indentation modulus maps at the junction between two adjacent facets of a tellurium microcrystal. The clear contrast observed in the elastic moduli of the two facets indicates the different surface crystallography of these facets.

  10. Fluorescence spectral correlation spectroscopy (FSCS) for probes with highly overlapping emission spectra

    Benda, A.; Kapusta, Peter; Hof, Martin; Gaus, K.

    2014-01-01

    Roč. 22, č. 3 (2014), s. 2973-2988 ISSN 1094-4087 R&D Projects: GA AV ČR KJB400400904; GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : spectroscopy * fluorescence and luminiscence * confocal microscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.488, year: 2014

  11. Discrimination of different red wine by Fourier-transform infrared and two-dimensional infrared correlation spectroscopy

    Zhang, Yan-ling; Chen, Jian-bo; Lei, Yu; Zhou, Qun; Sun, Su-qin; Noda, Isao

    2010-06-01

    Fourier-transform infrared spectroscopy (FT-IR) and two-dimensional infrared (2D IR) correlation spectroscopy were applied to analyze main components of liquid red wine with different sugar contents and volatilization residues of dry red wine from different manufactures. The infrared spectra, second derivative spectra of dry red wine show the typical peaks of alcohol, while the spectra of sweet wine are composed of the peaks of both alcohol and sugar, and the contribution of sugar enhanced as the increase of sugar content. Using principal component analysis (PCA) method, dry and sweet wine can be readily classified. Analysis of the infrared spectra of the volatilization residues of dry red wine samples from five different manufactures indicates that dry red wine may be composed of glycerol, carboxylic acids or esters and carboxyl ate, at the same time, different dry red wine show different characteristic peaks in the second derivative spectra and 2D IR correlation spectra, which can be used to discriminate the different manufactures and evaluate the quality of wine samples. The results suggested that infrared spectroscopy is a direct and effective method for the analysis of principle components of different red wines and discrimination of different red wines.

  12. Dipole and Coulomb forces in electron capture dissociation and electron transfer dissociation mass spectroscopy.

    Świerszcz, Iwona; Skurski, Piotr; Simons, Jack

    2012-02-23

    Ab initio electronic structure calculations were performed on a doubly charged polypeptide model H(+)-Lys(Ala)(19)-CO-CH(NH(2))-CH(2)-SS-CH(2)-(NH(2))CH-CO-(Ala)(19)-Lys-H(+) consisting of a C-terminal protonated Lys followed by a 19-Ala α-helix with a 20th Ala-like unit whose side chain is linked by a disulfide bond to a corresponding Ala-like unit connected to a second 19-Ala α-helix terminated by a second C-terminal-protonated Lys. The Coulomb potentials arising from the two charged Lys residues and dipole potentials arising from the two oppositely directed 72 D dipoles of the α-helices act to stabilize the SS bond's σ* orbital. The Coulomb potentials provide stabilization of 1 eV, while the two large dipoles generate an additional 4 eV. Such stabilization allows the SS σ* orbital to attach an electron and thereby generate disulfide bond cleavage products. Although calculations are performed only on SS bond cleavage, discussion of N-C(α) bond cleavage caused by electron attachment to amide π* orbitals is also presented. The magnitudes of the stabilization energies as well as the fact that they arise from Coulomb and dipole potentials are supported by results on a small model system consisting of a H(3)C-SS-CH(3) molecule with positive and negative fractional point charges to its left and right designed to represent (i) two positive charges ca. 32 Å distant (i.e., the two charged Lys sites of the peptide model) and (ii) two 72 D dipoles (i.e., the two α-helices). Earlier workers suggested that internal dipole forces in polypeptides could act to guide incoming free electrons (i.e., in electron capture dissociation (ECD)) toward the positive end of the dipole and thus affect the branching ratios for cleaving various bonds. Those workers argued that, because of the huge mass difference between an anion donor and a free electron, internal dipole forces would have a far smaller influence over the trajectory of a donor (i.e., in electron transfer dissociation

  13. Nanometrology of Biomass for Bioenergy: The Role of Atomic Force Microscopy and Spectroscopy in Plant Cell Characterization

    Anne M. Charrier

    2018-03-01

    Full Text Available Ethanol production using extracted cellulose from plant cell walls (PCW is a very promising approach to biofuel production. However, efficient throughput has been hindered by the phenomenon of recalcitrance, leading to high costs for the lignocellulosic conversion. To overcome recalcitrance, it is necessary to understand the chemical and structural properties of the plant biological materials, which have evolved to generate the strong and cohesive features observed in plants. Therefore, tools and methods that allow the investigation of how the different molecular components of PCW are organized and distributed and how this impacts the mechanical properties of the plants are needed but challenging due to the molecular and morphological complexity of PCW. Atomic force microscopy (AFM, capitalizing on the interfacial nanomechanical forces, encompasses a suite of measurement modalities for nondestructive material characterization. Here, we present a review focused on the utilization of AFM for imaging and determination of physical properties of plant-based specimens. The presented review encompasses the AFM derived techniques for topography imaging (AM-AFM, mechanical properties (QFM, and surface/subsurface (MSAFM, HPFM chemical composition imaging. In particular, the motivation and utility of force microscopy of plant cell walls from the early fundamental investigations to achieve a better understanding of the cell wall architecture, to the recent studies for the sake of advancing the biofuel research are discussed. An example of delignification protocol is described and the changes in morphology, chemical composition and mechanical properties and their correlation at the nanometer scale along the process are illustrated.

  14. Photon correlation spectroscopy of classical and non-classical light fields and its debt to Glauber and Harvard

    Pike, E.R.

    1985-01-01

    The Union Gikon Company of Japan has designed their new photon-correlation spectrometer to the outside world with the aid of a number of Japanese Universities and Industrial Research Laboratories. It comes with a list of some two dozen Japanese Institutions who have been using it successfully since its recent launch in their home country. The cost of the system is some $60,000 and it will no doubt, find its place in the market alongside existing well-known photon correlation systems such as those from Brookhaven Instruments Corporation, Hiac Royco, Coulter and Malvern Instruments. Although they may be used for other purposes, the main application of the instruments is the measurement of sizes of submicron particles such as proteins, enzymes, viruses, polymers and numerous other macro-molecular substances. A topical proposal, for example, is the detection of the immunological reactions of the AIDS virus. Photon correlation spectroscopy has become an important technique in modern laboratory practice

  15. Correlated single-crystal electronic absorption spectroscopy and X-ray crystallography at NSLS beamline X26-C

    Orville, A.M.; Buono, R.; Cowan, M.; Heroux, A.; Shea-McCarthy, G.; Schneider, D.K.; Skinner, J.M.; Skinner, M.J.; Stoner-Ma, D.; Sweet, R.M.

    2011-01-01

    The research philosophy and new capabilities installed at NSLS beamline X26-C to support electronic absorption and Raman spectroscopies coupled with X-ray diffraction are reviewed. This beamline is dedicated full time to multidisciplinary studies with goals that include revealing the relationship between the electronic and atomic structures in macromolecules. The beamline instrumentation has been fully integrated such that optical absorption spectra and X-ray diffraction images are interlaced. Therefore, optical changes induced by X-ray exposure can be correlated with X-ray diffraction data collection. The installation of Raman spectroscopy into the beamline is also briefly reviewed. Data are now routinely generated almost simultaneously from three complementary types of experiments from the same sample. The beamline is available now to the NSLS general user population.

  16. Combined multi-distance frequency domain and diffuse correlation spectroscopy system with simultaneous data acquisition and real-time analysis.

    Carp, Stefan A; Farzam, Parisa; Redes, Norin; Hueber, Dennis M; Franceschini, Maria Angela

    2017-09-01

    Frequency domain near infrared spectroscopy (FD-NIRS) and diffuse correlation spectroscopy (DCS) have emerged as synergistic techniques for the non-invasive assessment of tissue health. Combining FD-NIRS oximetry with DCS measures of blood flow, the tissue oxygen metabolic rate can be quantified, a parameter more closely linked to underlying physiology and pathology than either NIRS or DCS estimates alone. Here we describe the first commercially available integrated instrument, called the "MetaOx", designed to enable simultaneous FD-NIRS and DCS measurements at rates of 10 + Hz, and offering real-time data evaluation. We show simultaneously acquired characterization data demonstrating performance equivalent to individual devices and sample in vivo measurements of pulsation resolved blood flow, forearm occlusion hemodynamic changes and muscle oxygen metabolic rate monitoring during stationary bike exercise.

  17. Relationship between occlusal force and falls among community-dwelling elderly in Japan: a cross-sectional correlative study.

    Eto, Maki; Miyauchi, Shinji

    2018-05-09

    Falls may cause serious health conditions among older population. Fall-related physical factors are thought to be associated with occlusal conditions. However, few studies examined the relationship between occlusal force and falls. To identify the association between occlusal force and falls among community-dwelling elderly individuals in Japan, public health nurses conducted a cross-sectional descriptive study. We performed extensive physical assessments of five items: maximum occlusal force, handgrip strength, maximal knee extensor strength, one-leg standing time with eyes open and body sway. We also conducted a questionnaire survey concerning the participants' demographic characteristics, health status and fall experience during the past year. Mean scores and standard deviations were calculated for age and the total points of the index of activities of daily living. Associations were examined using Mann-Whitney tests and logistic regression. We examined 159 community-dwelling people aged ≥65 years, who were independent and active, including 38 participants (24.5%) with experience of falls in the past year. Maximum occlusal force had significant correlation with handgrip strength, maximal knee extensor strength, and one-leg standing time and body sway (P force (P = 0.004). This is the first study, led by public health nursing researchers, to examine the associations between maximum occlusal force and falls among community-dwelling elderly in Japan. The results showed that maximum occlusal force was significantly related to the other four extensive physical assessments, and might also suggest that maximum occlusal force assessment by public health nurses could contribute to more sophisticated and precise prediction of fall risks among the community-dwelling elderly. The latest occlusal force measurement device is non-invasive and easy to use. Public health nurses can introduce it at periodical community health checkup assembly events, which might

  18. Atmospheric Transmission Measurements Using IR Lasers, Fourier Transform Spectroscopy, and Gas-Filter Correlation Techniques

    Dowling, J

    1977-01-01

    ... and a gas filter correlation spectrometer. Results obtained from three concurrent experiments used to generate a data base appropriate to high resolution transmission model validation are displayed...

  19. Recent Developments in Fluorescence Correlation Spectroscopy for Diffusion Measurements in Planar Lipid Membranes

    Macháň, Radek; Hof, Martin

    2010-01-01

    Roč. 11, č. 2 (2010), s. 427-457 E-ISSN 1422-0067 R&D Projects: GA ČR GA203/08/0114; GA AV ČR GEMEM/09/E006 Institutional research plan: CEZ:AV0Z40400503 Keywords : lateral diffusion * fluorescence fluctuation spectroscopy * confocal microscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.279, year: 2010

  20. Fluorescence lifetime correlation spectroscopy combined with lifetime tuning: New perspectives in supported phospholipid bilayer research

    Benda, Aleš; Fagulová, Veronika; Deyneka, Alexander; Enderlain, J.; Hof, Martin

    2006-01-01

    Roč. 22, č. 23 (2006), s. 9580-9585 ISSN 0743-7463 R&D Projects: GA ČR GA203/05/2308; GA MŠk LC06063 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z10100522 Keywords : spectroscopy * fluorescence * FLCS Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.902, year: 2006

  1. On the use of band-target entropy minimization to simplify the interpretation of two-dimensional correlation spectroscopy.

    Widjaja, Effendi; Tan, Boon Hong; Garland, Marc

    2006-03-01

    Two-dimensional (2D) correlation spectroscopy has been extensively applied to analyze various vibrational spectroscopic data, especially infrared and Raman. However, when it is applied to real-world experimental data, which often contains various imperfections (such as noise interference, baseline fluctuations, and band-shifting) and highly overlapping bands, many artifacts and misleading features in synchronous and asynchronous maps will emerge, and this will lead to difficulties with interpretation. Therefore, an approach that counters many artifacts and therefore leads to simplified interpretation of 2D correlation analysis is certainly useful. In the present contribution, band-target entropy minimization (BTEM) is employed as a spectral pretreatment to handle many of the artifact problems before the application of 2D correlation analysis. BTEM is employed to elucidate the pure component spectra of mixtures and their corresponding concentration profiles. Two alternate forms of analysis result. In the first, the normally vxv problem is converted to an equivalent nvxnv problem, where n represents the number of species present. In the second, the pure component spectra are transformed into simple distributions, and an equivalent and less computationally intensive nv'xnv' problem results (v'evaporation study where in situ Fourier transform infrared (FT-IR) spectroscopy is used as the analytical tool.

  2. Force spectroscopy measurements show that cortical neurons exposed to excitotoxic agonists stiffen before showing evidence of bleb damage.

    Shan Zou

    Full Text Available In ischemic and traumatic brain injury, hyperactivated glutamate (N-methyl-D-aspartic acid, NMDA and sodium (Nav channels trigger excitotoxic neuron death. Na(+, Ca(++ and H2O influx into affected neurons elicits swelling (increased cell volume and pathological blebbing (disassociation of the plasma membrane's bilayer from its spectrin-actomyosin matrix. Though usually conflated in injured tissue, cell swelling and blebbing are distinct processes. Around an injury core, salvageable neurons could be mildly swollen without yet having suffered the bleb-type membrane damage that, by rendering channels leaky and pumps dysfunctional, exacerbates the excitotoxic positive feedback spiral. Recognizing when neuronal inflation signifies non-lethal osmotic swelling versus blebbing should further efforts to salvage injury-penumbra neurons. To assess whether the mechanical properties of osmotically-swollen versus excitotoxically-blebbing neurons might be cytomechanically distinguishable, we measured cortical neuron elasticity (gauged via atomic force microscopy (AFM-based force spectroscopy upon brief exposure to hypotonicity or to excitotoxic agonists (glutamate and Nav channel activators, NMDA and veratridine. Though unperturbed by solution exchange per se, elasticity increased abruptly with hypotonicity, with NMDA and with veratridine. Neurons then invariably softened towards or below the pre-treatment level, sometimes starting before the washout. The initial channel-mediated stiffening bespeaks an abrupt elevation of hydrostatic pressure linked to NMDA or Nav channel-mediated ion/H2O fluxes, together with increased [Ca(++]int-mediated submembrane actomyosin contractility. The subsequent softening to below-control levels is consistent with the onset of a lethal level of bleb damage. These findings indicate that dissection/identification of molecular events during the excitotoxic transition from stiff/swollen to soft/blebbing is warranted and should be

  3. Lipid Bilayer Membrane in a Silicon Based Micron Sized Cavity Accessed by Atomic Force Microscopy and Electrochemical Impedance Spectroscopy.

    Khan, Muhammad Shuja; Dosoky, Noura Sayed; Patel, Darayas; Weimer, Jeffrey; Williams, John Dalton

    2017-07-05

    Supported lipid bilayers (SLBs) are widely used in biophysical research to probe the functionality of biological membranes and to provide diagnoses in high throughput drug screening. Formation of SLBs at below phase transition temperature ( Tm ) has applications in nano-medicine research where low temperature profiles are required. Herein, we report the successful production of SLBs at above-as well as below-the Tm of the lipids in an anisotropically etched, silicon-based micro-cavity. The Si-based cavity walls exhibit controlled temperature which assist in the quick and stable formation of lipid bilayer membranes. Fusion of large unilamellar vesicles was monitored in real time in an aqueous environment inside the Si cavity using atomic force microscopy (AFM), and the lateral organization of the lipid molecules was characterized until the formation of the SLBs. The stability of SLBs produced was also characterized by recording the electrical resistance and the capacitance using electrochemical impedance spectroscopy (EIS). Analysis was done in the frequency regime of 10 -2 -10⁵ Hz at a signal voltage of 100 mV and giga-ohm sealed impedance was obtained continuously over four days. Finally, the cantilever tip in AFM was utilized to estimate the bilayer thickness and to calculate the rupture force at the interface of the tip and the SLB. We anticipate that a silicon-based, micron-sized cavity has the potential to produce highly-stable SLBs below their Tm . The membranes inside the Si cavity could last for several days and allow robust characterization using AFM or EIS. This could be an excellent platform for nanomedicine experiments that require low operating temperatures.

  4. AFM force spectroscopy reveals how subtle structural differences affect the interaction strength between Candida albicans and DC-SIGN.

    te Riet, Joost; Reinieren-Beeren, Inge; Figdor, Carl G; Cambi, Alessandra

    2015-11-01

    The fungus Candida albicans is the most common cause of mycotic infections in immunocompromised hosts. Little is known about the initial interactions between Candida and immune cell receptors, such as the C-type lectin dendritic cell-specific intracellular cell adhesion molecule-3 (ICAM-3)-grabbing non-integrin (DC-SIGN), because a detailed characterization at the structural level is lacking. DC-SIGN recognizes specific Candida-associated molecular patterns, that is, mannan structures present in the cell wall of Candida. The molecular recognition mechanism is however poorly understood. We postulated that small differences in mannan-branching may result in considerable differences in the binding affinity. Here, we exploit atomic force microscope-based dynamic force spectroscopy with single Candida cells to gain better insight in the carbohydrate recognition capacity of DC-SIGN. We demonstrate that slight differences in the N-mannan structure of Candida, that is, the absence or presence of a phosphomannan side chain, results in differences in the recognition by DC-SIGN as follows: (i) it contributes to the compliance of the outer cell wall of Candida, and (ii) its presence results in a higher binding energy of 1.6 kB T. The single-bond affinity of tetrameric DC-SIGN for wild-type C. albicans is ~10.7 kB T and a dissociation constant kD of 23 μM, which is relatively strong compared with other carbohydrate-protein interactions described in the literature. In conclusion, this study shows that DC-SIGN specifically recognizes mannan patterns on C. albicans with high affinity. Knowledge on the binding pocket of DC-SIGN and its pathogenic ligands will lead to a better understanding of how fungal-associated carbohydrate structures are recognized by receptors of the immune system and can ultimately contribute to the development of new anti-fungal drugs. Copyright © 2015 John Wiley & Sons, Ltd.

  5. Study of the mixed alkali effect in lithium and sodium metaphosphate glass-forming liquids by photon correlation spectroscopy

    Changstrom, J R; Sidebottom, D L

    2008-01-01

    We report results of an extensive study of the structural relaxation occurring in mixed alkali metaphosphate liquids obtained by photon correlation spectroscopy. Values for the glass transition temperature, the fragility index, and the heterogeneity parameter (also known as the Kohlrausch exponent) are extracted from the measurements and are all shown to exhibit a mixed alkali effect wherein nonlinear variations with mixing occur. The depression in the glass transition temperature is shown to be the direct result of mechanical relaxations, present in the solid, which prematurely loosen the glass structure. A minimum in the fragility index is believed to be an artifact of the resulting depression of the glass transition temperature

  6. Membrane mobility and microdomain association of the dopamine transporter studied with fluorescence correlation spectroscopy and fluorescence recovery after photobleaching

    Adkins, Erika M; Samuvel, Devadoss J; Fog, Jacob U

    2007-01-01

    To investigate microdomain association of the dopamine transporter (DAT), we employed FCS (fluorescence correlation spectroscopy) and FRAP (fluorescence recovery after photobleaching). In non-neuronal cells (HEK293), FCS measurements revealed for the YFP-DAT (DAT tagged with yellow fluorescent...... protein) a diffusion coefficient (D) of approximately 3.6 x 10(-9) cm2/s, consistent with a relatively freely diffusible protein. In neuronally derived cells (N2a), we were unable to perform FCS measurements on plasma membrane-associated protein due to photobleaching, suggesting partial immobilization...

  7. Membrane mobility and microdomain association of the dopaminetransporter studied with fluorescence correlation spectroscopy and fluorescence recovery after photobleaching

    Adkins, Erika; Samuvel, Devadoss; Fog, Jacob

    2007-01-01

    To investigate microdomain association of the dopamine transporter (DAT), we employed FCS (fluorescence correlation spectroscopy) and FRAP (fluorescence recovery after photobleaching). In non-neuronal cells (HEK293), FCS measurements revealed for the YFP-DAT (DAT tagged with yellow fluorescent...... protein) a diffusion coefficient (D) of ~3.6 × 10-9 cm2/s, consistent with a relatively freely diffusible protein. In neuronally derived cells (N2a), we were unable to perform FCS measurements on plasma membrane-associated protein due to photobleaching, suggesting partial immobilization...

  8. Investigation of free fatty acid associated recombinant membrane receptor protein expression in HEK293 cells using Raman spectroscopy, calcium imaging, and atomic force microscopy.

    Lin, Juqiang; Xu, Han; Wu, Yangzhe; Tang, Mingjie; McEwen, Gerald D; Liu, Pin; Hansen, Dane R; Gilbertson, Timothy A; Zhou, Anhong

    2013-02-05

    G-protein-coupled receptor 120 (GPR120) is a previously orphaned G-protein-coupled receptor that apparently functions as a sensor for dietary fat in the gustatory and digestive systems. In this study, a cDNA sequence encoding a doxycycline (Dox)-inducible mature peptide of GPR120 was inserted into an expression vector and transfected in HEK293 cells. We measured Raman spectra of single HEK293 cells as well as GPR120-expressing HEK293-GPR120 cells at a 48 h period following the additions of Dox at several concentrations. We found that the spectral intensity of HEK293-GPR120 cells is dependent upon the dose of Dox, which correlates with the accumulation of GPR120 protein in the cells. However, the amount of the fatty acid activated changes in intracellular calcium (Ca(2+)) as measured by ratiometric calcium imaging was not correlated with Dox concentration. Principal components analysis (PCA) of Raman spectra reveals that the spectra from different treatments of HEK293-GPR120 cells form distinct, completely separated clusters with the receiver operating characteristic (ROC) area of 1, while those spectra for the HEK293 cells form small overlap clusters with the ROC area of 0.836. It was also found that expression of GPR120 altered the physiochemical and biomechanical properties of the parental cell membrane surface, which was quantitated by atomic force microscopy (AFM). These findings demonstrate that the combination of Raman spectroscopy, calcium imaging, and AFM may provide new tools in noninvasive and quantitative monitoring of membrane receptor expression induced alterations in the biophysical and signaling properties of single living cells.

  9. MDM2–MDM4 molecular interaction investigated by atomic force spectroscopy and surface plasmon resonance

    Moscetti, Ilaria; Teveroni, Emanuela; Moretti, Fabiola; Bizzarri, Anna Rita; Cannistraro, Salvatore

    2016-01-01

    Murine double minute 2 (MDM2) and 4 (MDM4) are known as the main negative regulators of p53, a tumor suppressor. They are able to form heterodimers that are much more effective in the downregulation of p53. Therefore, the MDM2–MDM4 complex could be a target for promising therapeutic restoration of p53 function. To this aim, a deeper understanding of the molecular mechanisms underlining the heterodimerization is needed. The kinetic and thermodynamic characterization of the MDM2–MDM4 complex was performed with two complementary approaches: atomic force spectroscopy and surface plasmon resonance. Both techniques revealed an equilibrium dissociation constant (KD) in the micromolar range for the MDM2–MDM4 heterodimer, similar to related complexes involved in the p53 network. Furthermore, the MDM2–MDM4 complex is characterized by a relatively high free energy, through a single energy barrier, and by a lifetime in the order of tens of seconds. New insights into the MDM2–MDM4 interaction could be highly important for developing innovative anticancer drugs focused on p53 reactivation. PMID:27621617

  10. Medulloblastoma: correlation among findings of conventional magnetic resonance imaging, diffusion-weighted imaging and proton magnetic resonance spectroscopy

    Fonte, Mariana Vieira de Melo da; Otaduy, Maria Concepcion Garcia; Lucato, Leandro Tavares; Reed, Umbertina Conti; Leite, Claudia da Costa [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Hospital das Clinicas. Inst. de Radiologia]. E-mail: mvmfonte@uol.com.br; Costa, Maria Olivia Rodrigues; Amaral, Raquel Portugal Guimaraes [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Faculdade de Medicina. Dept. de Radiologia; Reed, Umbertina Conti [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Faculdade de Medicina. Dept. de Neurologia; Rosemberg, Sergio [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Hospital das Clinicas. Dept. de Patologia

    2008-11-15

    To correlate imaging findings of medulloblastomas at conventional magnetic resonance imaging, diffusion-weighted imaging and proton magnetic resonance spectroscopy, comparing them with data in the literature. Preoperative magnetic resonance imaging studies of nine pediatric patients with histologically confirmed medulloblastomas (eight desmoplastic medulloblastoma, and one giant cell medulloblastoma) were retrospectively reviewed, considering demographics as well as tumors characteristics such as localization, morphology, signal intensity, contrast-enhancement, dissemination, and diffusion-weighted imaging and spectroscopy findings. In most of cases the tumors were centered in the cerebellar vermis (77.8%), predominantly solid (88.9%), hypointense on T 1-weighted images and intermediate/hyperintense on T 2-FLAIR-weighted images, with heterogeneous enhancement (100%), tumor dissemination/extension (77.8%) and limited water molecule mobility (100%). Proton spectroscopy acquired with STEAM technique (n = 6) demonstrated decreased Na a / Cr ratio (83.3%) and increased Co/Cr (100%) and ml/Cr (66.7%) ratios; and with PRESS technique (n = 7) demonstrated lactate peak (57.1%). Macroscopic magnetic resonance imaging findings in association with biochemical features of medulloblastomas have been useful in the differentiation among the most frequent posterior fossa tumors. (author)

  11. Medulloblastoma: correlation among findings of conventional magnetic resonance imaging, diffusion-weighted imaging and proton magnetic resonance spectroscopy

    Fonte, Mariana Vieira de Melo da; Otaduy, Maria Concepcion Garcia; Lucato, Leandro Tavares; Reed, Umbertina Conti; Leite, Claudia da Costa; Costa, Maria Olivia Rodrigues; Amaral, Raquel Portugal Guimaraes; Reed, Umbertina Conti; Rosemberg, Sergio

    2008-01-01

    To correlate imaging findings of medulloblastomas at conventional magnetic resonance imaging, diffusion-weighted imaging and proton magnetic resonance spectroscopy, comparing them with data in the literature. Preoperative magnetic resonance imaging studies of nine pediatric patients with histologically confirmed medulloblastomas (eight desmoplastic medulloblastoma, and one giant cell medulloblastoma) were retrospectively reviewed, considering demographics as well as tumors characteristics such as localization, morphology, signal intensity, contrast-enhancement, dissemination, and diffusion-weighted imaging and spectroscopy findings. In most of cases the tumors were centered in the cerebellar vermis (77.8%), predominantly solid (88.9%), hypointense on T 1-weighted images and intermediate/hyperintense on T 2-FLAIR-weighted images, with heterogeneous enhancement (100%), tumor dissemination/extension (77.8%) and limited water molecule mobility (100%). Proton spectroscopy acquired with STEAM technique (n = 6) demonstrated decreased Na a / Cr ratio (83.3%) and increased Co/Cr (100%) and ml/Cr (66.7%) ratios; and with PRESS technique (n = 7) demonstrated lactate peak (57.1%). Macroscopic magnetic resonance imaging findings in association with biochemical features of medulloblastomas have been useful in the differentiation among the most frequent posterior fossa tumors. (author)

  12. Easy measurement of diffusion coefficients of EGFP-tagged plasma membrane proteins using k-space Image Correlation Spectroscopy

    Christensen, Eva Arnspang; Koffman, Jennifer Skaarup; Marlar, Saw

    2014-01-01

    Lateral diffusion and compartmentalization of plasma membrane proteins are tightly regulated in cells and thus, studying these processes will reveal new insights to plasma membrane protein function and regulation. Recently, k-Space Image Correlation Spectroscopy (kICS)1 was developed to enable...... routine measurements of diffusion coefficients directly from images of fluorescently tagged plasma membrane proteins, that avoided systematic biases introduced by probe photophysics. Although the theoretical basis for the analysis is complex, the method can be implemented by nonexperts using a freely...... to the correlation function yields the diffusion coefficient. This paper provides a step-by-step guide to the image analysis and measurement of diffusion coefficients via kICS. First, a high frame rate image sequence of a fluorescently labeled plasma membrane protein is acquired using a fluorescence microscope Then...

  13. A method of mössbauer fourier spectroscopy for determination of the biopolimer coordinate correlation functions

    Basovets, S. K.; Uporov, I. V.; Shaitan, K. V.; Krupyanskii, Yu. F.; Kurinov, I. V.; Suzdalev, I. P.; Rubin, A. B.; Goldanskii, V. I.

    1988-12-01

    A method of Mössbauer Fourier spectroscopy is developed to determine the correlation function of coordinates of a macromolecular system. The method does not require the use of an a priori dynamic model. The application of the method to the analysis of RSMR data for human serum albumin has demonstrated considerable changes in the dynamic behavior of the protein globule when the temperature is changed from 270 to 310 K. The main conclusions of the present work is the simultaneous observation of low-frequency (τ≥10-9 sec) and high-frequency (τ≪10-9 sec) large-scaled motions, that is the two-humped distribution of correlation times of protein motions.

  14. Quantifying the number of color centers in single fluorescent nanodiamonds by photon correlation spectroscopy and Monte Carlo simulation

    Hui, Y.Y.; Chang, Y.-R.; Lee, H.-Y.; Chang, H.-C.; Lim, T.-S.; Fann Wunshain

    2009-01-01

    The number of negatively charged nitrogen-vacancy centers (N-V) - in fluorescent nanodiamond (FND) has been determined by photon correlation spectroscopy and Monte Carlo simulations at the single particle level. By taking account of the random dipole orientation of the multiple (N-V) - fluorophores and simulating the probability distribution of their effective numbers (N e ), we found that the actual number (N a ) of the fluorophores is in linear correlation with N e , with correction factors of 1.8 and 1.2 in measurements using linearly and circularly polarized lights, respectively. We determined N a =8±1 for 28 nm FND particles prepared by 3 MeV proton irradiation

  15. Anharmonic Infrared Spectroscopy through the Fourier Transform of Time Correlation Function Formalism in ONETEP.

    Vitale, Valerio; Dziedzic, Jacek; Dubois, Simon M-M; Fangohr, Hans; Skylaris, Chris-Kriton

    2015-07-14

    Density functional theory molecular dynamics (DFT-MD) provides an efficient framework for accurately computing several types of spectra. The major benefit of DFT-MD approaches lies in the ability to naturally take into account the effects of temperature and anharmonicity, without having to introduce any ad hoc or a posteriori corrections. Consequently, computational spectroscopy based on DFT-MD approaches plays a pivotal role in the understanding and assignment of experimental peaks and bands at finite temperature, particularly in the case of floppy molecules. Linear-scaling DFT methods can be used to study large and complex systems, such as peptides, DNA strands, amorphous solids, and molecules in solution. Here, we present the implementation of DFT-MD IR spectroscopy in the ONETEP linear-scaling code. In addition, two methods for partitioning the dipole moment within the ONETEP framework are presented. Dipole moment partitioning allows us to compute spectra of molecules in solution, which fully include the effects of the solvent, while at the same time removing the solvent contribution from the spectra.

  16. Superatom spectroscopy and the electronic state correlation between elements and isoelectronic molecular counterparts.

    Peppernick, Samuel J; Gunaratne, K D Dasitha; Castleman, A W

    2010-01-19

    Detailed in the present investigation are results pertaining to the photoelectron spectroscopy of negatively charged atomic ions and their isoelectronic molecular counterparts. Experiments utilizing the photoelectron imaging technique are performed on the negative ions of the group 10 noble metal block (i.e. Ni-, Pd-, and Pt-) of the periodic table at a photon energy of 2.33 eV (532 nm). The accessible electronic transitions, term energies, and orbital angular momentum components of the bound electronic states in the atom are then compared with photoelectron images collected for isoelectronic early transition metal heterogeneous diatomic molecules, M-X- (M = Ti,Zr,W; X = O or C). A superposition principle connecting the spectroscopy between the atomic and molecular species is observed, wherein the electronic structure of the diatomic is observed to mimic that present in the isoelectronic atom. The molecular ions studied in this work, TiO-, ZrO-, and WC- can then be interpreted as possessing superatomic electronic structures reminiscent of the isoelectronic elements appearing on the periodic table, thereby quantifying the superatom concept.

  17. [Constitutive correlations for wire-wrapped subchannel analysis under forced and mixed convection conditions]. Part II

    Cheng, S.K.; Todreas, N.E.

    1984-08-01

    A new version of the ENERGY series code, ENERGY-IV, was written for predicting coolant temperature distributions in wire-wrapped rod assemblies used in the Liquid Metal Fast Breeder Reactor. The ENERGY-IV Code is applicable to both steady-state forced and mixed convection operation for a single isolated assembly. (The SUPERENERGY Code, [Basehore (1980)] is applicable to core wide forced convection analysis.) ENERGY-IV is an empirical code designed to be fast running. Hence the core designer can use it as an inexpensive thermal hydraulic design or diagnosis tool

  18. 2D FTIR correlation spectroscopy and EPR analysis of Urtica dioica leaves from areas of different environmental pollution.

    Moskal, Paulina; Wesełucha-Birczyńska, Aleksandra; Łabanowska, Maria; Kurdziel, Magdalena; Filek, Maria

    2018-01-15

    Leaves of Urtica dioica collected from two areas of different environmental pollution were analysed by fourier transform infrared spectroscopy (FTIR) and electron paramagnetic resonance (EPR) spectroscopy. Analysis of FTIR spectra allows to describe main component of plant like proteins, lipids and carbohydrates. Although the FTIR spectra of plants from these two geographical locations of different environmental pollution appear to be relatively similar, 2D correlation shows completely different patterns. Synchronous and asynchronous correlation maps showed sequences of changes occurring during development of plant, manly in Amide I and Amide II, lignin, lipids and cellulose. In addition, 2D analysis revealed another sequence of changes as the function of plant growth depending on the degree of the environmental pollution. Two various kinds of paramagnetic species, transition metal ions (Mn(II), Fe(III)) and stable organic radicals (chlorophyll, semiquinone, tyrosyl and carbon centered) were found in leaves of nettle collected at different stages of development and growing in clean and polluted environment. In plants growing in polluted area the injuries of protein molecules bonding metal ions and the disturbances of photosynthesis and redox equilibrium in cells, as well as instability of polysaccharide structure of cell walls were observed. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. A comparison of single particle tracking and temporal image correlation spectroscopy for quantitative analysis of endosome motility

    Lund, F. W.; Wustner, D.

    2013-01-01

    Single particle tracking (SPT) is becoming a standard method to extract transport parameters from time-lapse image sequences of fluorescent vesicles in living cells. Another method to obtain these data is temporal image correlation spectroscopy (TICS), but this method is less often used for measu......Single particle tracking (SPT) is becoming a standard method to extract transport parameters from time-lapse image sequences of fluorescent vesicles in living cells. Another method to obtain these data is temporal image correlation spectroscopy (TICS), but this method is less often used...... for measurement of intracellular vesicle transport. Here, we present an extensive comparison of SPT and TICS. First we examine the effect of photobleaching, shading and noise on SPT and TICS analysis using simulated image sequences. To this end, we developed a simple photophysical model, which relates spatially...... varying illumination intensity to the bleaching propensity and fluorescence intensity of the moving particles. We found that neither SPT nor TICS are affected by photobleaching per se, but the transport parameters obtained by both methods are sensitive to the signal-to-noise ratio. In addition, the number...

  20. Quantitative HRMAS proton total correlation spectroscopy applied to cultured melanoma cells treated by chloroethyl nitrosourea: demonstration of phospholipid metabolism alterations.

    Morvan, Daniel; Demidem, Aicha; Papon, Janine; Madelmont, Jean Claude

    2003-02-01

    Recent NMR spectroscopy developments, such as high-resolution magic angle spinning (HRMAS) probes and correlation-enhanced 2D sequences, now allow improved investigations of phospholipid (Plp) metabolism. Using these modalities we previously demonstrated that a mouse-bearing melanoma tumor responded to chloroethyl nitrosourea (CENU) treatment in vivo by altering its Plp metabolism. The aims of the present study were to investigate whether HRMAS proton total correlation spectroscopy (TOCSY) could be used as a quantitative technique to probe Plp metabolism, and to determine the Plp metabolism response of cultured B16 melanoma cells to CENU treatment in vitro. The exploited TOCSY signals of Plp derivatives arose from scalar coupling among the protons of neighbor methylene groups within base headgroups (choline and ethanolamine). For strongly expressed Plp derivatives, TOCSY signals were compared to saturation recovery signals and demonstrated a linear relationship. HRMAS proton TOCSY was thus used to provide concentrations of Plp derivatives during long-term follow-up of CENU-treated cell cultures. Strong Plp metabolism alteration was observed in treated cultured cells in vitro involving a down-regulation of phosphocholine, and a dramatic and irreversible increase of phosphoethanolamine. These findings are discussed in relation to previous in vivo data, and to Plp metabolism enzymatic involvement. Copyright 2003 Wiley-Liss, Inc.

  1. 2D FTIR correlation spectroscopy and EPR analysis of Urtica dioica leaves from areas of different environmental pollution

    Moskal, Paulina; Wesełucha-Birczyńska, Aleksandra; Łabanowska, Maria; Kurdziel, Magdalena; Filek, Maria

    2018-01-01

    Leaves of Urtica dioica collected from two areas of different environmental pollution were analysed by fourier transform infrared spectroscopy (FTIR) and electron paramagnetic resonance (EPR) spectroscopy. Analysis of FTIR spectra allows to describe main component of plant like proteins, lipids and carbohydrates. Although the FTIR spectra of plants from these two geographical locations of different environmental pollution appear to be relatively similar, 2D correlation shows completely different patterns. Synchronous and asynchronous correlation maps showed sequences of changes occurring during development of plant, manly in Amide I and Amide II, lignin, lipids and cellulose. In addition, 2D analysis revealed another sequence of changes as the function of plant growth depending on the degree of the environmental pollution. Two various kinds of paramagnetic species, transition metal ions (Mn(II), Fe(III)) and stable organic radicals (chlorophyll, semiquinone, tyrosyl and carbon centered) were found in leaves of nettle collected at different stages of development and growing in clean and polluted environment. In plants growing in polluted area the injuries of protein molecules bonding metal ions and the disturbances of photosynthesis and redox equilibrium in cells, as well as instability of polysaccharide structure of cell walls were observed.

  2. Vibrational correlation between conjugated carbonyl and diazo modes studied by single- and dual-frequency two-dimensional infrared spectroscopy

    Maekawa, Hiroaki; Sul, Soohwan; Ge, Nien-Hui

    2013-01-01

    Highlights: ► Vibrational dynamics of conjugated C=O and N=N modes of ethyl diazoacetate was studied. ► Their frequency–frequency correlation functions are different. ► The dual-frequency 2D IR spectrum indicates anticorrelated frequency fluctuations. ► Correlation effects on dual-frequency 2D IR spectra are discussed. ► The existence of cis and trans conformers is revealed in 2D IR spectra. - Abstract: We have applied infrared three-pulse photon echo and single- and dual-frequency 2D IR spectroscopy to the ester C=O and diazo N=N stretching modes in ethyl diazoacetate (EDA), and investigated their vibrational frequency fluctuations and correlation. The two modes exhibit different vibrational dynamics and 2D lineshape, which are well simulated by frequency–frequency correlation functions (FFCFs) with two decaying components. Although the FT IR spectrum shows a single C=O band, absolute magnitude 2D IR nonrephasing spectrum displays spectral signatures supporting the presence of cis and trans conformations. The cross-peak inclined toward the anti-diagonal in the dual-frequency 2D IR spectrum, indicating that the frequency fluctuations of the two modes are anticorrelated. This behavior is attributed to anticorrelated change in the bond orders when solvent and structural fluctuations causes EDA to adopt a different mixture of the two dominant resonance structures. The effects of cross FFCF on the cross-peak line shape are discussed

  3. Vibrational correlation between conjugated carbonyl and diazo modes studied by single- and dual-frequency two-dimensional infrared spectroscopy

    Maekawa, Hiroaki; Sul, Soohwan [Department of Chemistry, University of California at Irvine, Irvine, CA 92697-2025 (United States); Ge, Nien-Hui, E-mail: nhge@uci.edu [Department of Chemistry, University of California at Irvine, Irvine, CA 92697-2025 (United States)

    2013-08-30

    Highlights: ► Vibrational dynamics of conjugated C=O and N=N modes of ethyl diazoacetate was studied. ► Their frequency–frequency correlation functions are different. ► The dual-frequency 2D IR spectrum indicates anticorrelated frequency fluctuations. ► Correlation effects on dual-frequency 2D IR spectra are discussed. ► The existence of cis and trans conformers is revealed in 2D IR spectra. - Abstract: We have applied infrared three-pulse photon echo and single- and dual-frequency 2D IR spectroscopy to the ester C=O and diazo N=N stretching modes in ethyl diazoacetate (EDA), and investigated their vibrational frequency fluctuations and correlation. The two modes exhibit different vibrational dynamics and 2D lineshape, which are well simulated by frequency–frequency correlation functions (FFCFs) with two decaying components. Although the FT IR spectrum shows a single C=O band, absolute magnitude 2D IR nonrephasing spectrum displays spectral signatures supporting the presence of cis and trans conformations. The cross-peak inclined toward the anti-diagonal in the dual-frequency 2D IR spectrum, indicating that the frequency fluctuations of the two modes are anticorrelated. This behavior is attributed to anticorrelated change in the bond orders when solvent and structural fluctuations causes EDA to adopt a different mixture of the two dominant resonance structures. The effects of cross FFCF on the cross-peak line shape are discussed.

  4. Studying flow close to an interface by total internal reflection fluorescence cross-correlation spectroscopy: Quantitative data analysis

    Schmitz, R.; Yordanov, S.; Butt, H. J.; Koynov, K.; Dünweg, B.

    2011-12-01

    Total internal reflection fluorescence cross-correlation spectroscopy (TIR-FCCS) has recently [S. Yordanov , Optics ExpressOPEXFF1094-408710.1364/OE.17.021149 17, 21149 (2009)] been established as an experimental method to probe hydrodynamic flows near surfaces, on length scales of tens of nanometers. Its main advantage is that fluorescence occurs only for tracer particles close to the surface, thus resulting in high sensitivity. However, the measured correlation functions provide only rather indirect information about the flow parameters of interest, such as the shear rate and the slip length. In the present paper, we show how to combine detailed and fairly realistic theoretical modeling of the phenomena by Brownian dynamics simulations with accurate measurements of the correlation functions, in order to establish a quantitative method to retrieve the flow properties from the experiments. First, Brownian dynamics is used to sample highly accurate correlation functions for a fixed set of model parameters. Second, these parameters are varied systematically by means of an importance-sampling Monte Carlo procedure in order to fit the experiments. This provides the optimum parameter values together with their statistical error bars. The approach is well suited for massively parallel computers, which allows us to do the data analysis within moderate computing times. The method is applied to flow near a hydrophilic surface, where the slip length is observed to be smaller than 10nm, and, within the limitations of the experiments and the model, indistinguishable from zero.

  5. Feasibility of the simultaneous determination of polycyclic aromatic hydrocarbons based on two-dimensional fluorescence correlation spectroscopy

    Yang, Renjie; Dong, Guimei; Sun, Xueshan; Yang, Yanrong; Yu, Yaping; Liu, Haixue; Zhang, Weiyu

    2018-02-01

    A new approach for quantitative determination of polycyclic aromatic hydrocarbons (PAHs) in environment was proposed based on two-dimensional (2D) fluorescence correlation spectroscopy in conjunction with multivariate method. 40 mixture solutions of anthracene and pyrene were prepared in the laboratory. Excitation-emission matrix (EEM) fluorescence spectra of all samples were collected. And 2D fluorescence correlation spectra were calculated under the excitation perturbation. The N-way partial least squares (N-PLS) models were developed based on 2D fluorescence correlation spectra, showing a root mean square error of calibration (RMSEC) of 3.50 μg L- 1 and root mean square error of prediction (RMSEP) of 4.42 μg L- 1 for anthracene and of 3.61 μg L- 1 and 4.29 μg L- 1 for pyrene, respectively. Also, the N-PLS models were developed for quantitative analysis of anthracene and pyrene using EEM fluorescence spectra. The RMSEC and RMSEP were 3.97 μg L- 1 and 4.63 μg L- 1 for anthracene, 4.46 μg L- 1 and 4.52 μg L- 1 for pyrene, respectively. It was found that the N-PLS model using 2D fluorescence correlation spectra could provide better results comparing with EEM fluorescence spectra because of its low RMSEC and RMSEP. The methodology proposed has the potential to be an alternative method for detection of PAHs in environment.

  6. Hepatocarcinogenesis tumor grading correlated with in vivo image-guided 1H-NMR spectroscopy in a rat model

    Towner, Rheal A.; Foley, Lesley M.; Painter, Dorothy M.

    2005-01-01

    Hepatocellular carcinoma (HCC) is a common malignancy worldwide, the occurrence of which is unevenly distributed. Most hepatocellular carcinoma cases present late and have a poor prognosis; therefore, early diagnosis is essential to prolong survival. Differential diagnosis with magnetic resonance imaging (MRI) is difficult. We studied the feasibility of using magnetic resonance spectroscopy (MRS) at 7.0 T for the diagnosis and grading of liver tumors. An animal model of hepatocarcinogenesis was used, which allowed tumor progression from precancerous lesions to hepatocellular carcinomas. This study was focused primarily on the grading of the tumors and its correlation with the ratio between the MRS peaks arising from MRS-detected lipid hydrogens (0.9, 1.3 and 5.3 ppm) and compared to the γ-methylene hydrogens of glutamate (Glu) and glutamine (Gln) which was used as an internal reference (2.4 ppm). The lipid methylene hydrogen (1.3 ppm) to (Glu + Gln) ratio was found to correlate with the formation of differentiated small foci and (precancerous) hepatic nodules, whereas the unsaturated olefinic lipid hydrogen (5.3 ppm) to (Glu + Gln) ratio was able to correlate with the formation of late stage tumors such as adenomas and hepatocellular carcinomas. The results of our study suggest that MRS-detected alterations in lipid metabolism can be correlated with the grading of liver tumor tissue at different stages during the carcinogenesis process

  7. Correlated fermionic densities for many harmonically trapped particles interacting with repulsive forces

    Glasser, M.L.; March, N.H.; Nieto, L.M.

    2010-01-01

    This study is motivated by the very recent work on correlation energy as approximated by the Thomas-Fermi (TF) semiclassical limit [B.R. Landry, et al., Phys. Rev. Lett. 103 (2009) 066401]. In contrast, and motivated by the Hohenberg-Kohn theorem, our work is focussed primarily on the correlated TF ground-state density. We invoke directly the Holas et al. result that for two-fermion systems with harmonic trapping, the fermion-fermion interaction u simply adds to the trapping potential. We conclude this report with some results on correlation kinetic energy for two-fermion systems.

  8. Integration of Correlative Raman microscopy in a dual beam FIB-SEM J. of Raman Spectroscopy

    Timmermans, Frank Jan; Liszka, B.; Lenferink, Aufrid T.M.; van Wolferen, Hendricus A.G.M.; Otto, Cornelis

    2016-01-01

    We present an integrated confocal Raman microscope in a focused ion beam scanning electron microscope (FIB SEM). The integrated system enables correlative Raman and electron microscopic analysis combined with focused ion beam sample modification on the same sample location. This provides new

  9. Time dependence of the natural passivation process on AISI 304 in an alkaline medium: Atomic force microscopy and scanning Kelvin probe force microscopy as additional tools to electrochemical impedance spectroscopy

    Benaioun, N. E.; Maafa, I.; Florentin, A.; Denys, E.; Hakiki, N. E.; Moulayat, N.; Bubendorff, J. L.

    2018-04-01

    Thin surface films formed on AISI 304 samples in an alkaline solution of pH = 13 are studied by atomic force microscopy (AFM), scanning Kelvin probe force microscopy (SKPFM) and electrochemical impedance spectroscopy (EIS) as a function of immersion time. The results reveal that changes on EIS diagrams correspond to topographical modifications on the sample surface as shown by AFM. Both techniques are therefore complementary. The oxide layer is chemically homogenous as shown by SKPFM imaging and our ultra-thin passive layer is an efficient barrier against corrosion.

  10. Silicon nitride grids are compatible with correlative negative staining electron microscopy and tip-enhanced Raman spectroscopy for use in the detection of micro-organisms.

    Lausch, V; Hermann, P; Laue, M; Bannert, N

    2014-06-01

    Successive application of negative staining transmission electron microscopy (TEM) and tip-enhanced Raman spectroscopy (TERS) is a new correlative approach that could be used to rapidly and specifically detect and identify single pathogens including bioterrorism-relevant viruses in complex samples. Our objective is to evaluate the TERS-compatibility of commonly used electron microscopy (EM) grids (sample supports), chemicals and negative staining techniques and, if required, to devise appropriate alternatives. While phosphortungstic acid (PTA) is suitable as a heavy metal stain, uranyl acetate, paraformaldehyde in HEPES buffer and alcian blue are unsuitable due to their relatively high Raman scattering. Moreover, the low thermal stability of the carbon-coated pioloform film on copper grids (pioloform grids) negates their utilization. The silicon in the cantilever of the silver-coated atomic force microscope tip used to record TERS spectra suggested that Si-based grids might be employed as alternatives. From all evaluated Si-based TEM grids, the silicon nitride (SiN) grid was found to be best suited, with almost no background Raman signals in the relevant spectral range, a low surface roughness and good particle adhesion properties that could be further improved by glow discharge. Charged SiN grids have excellent particle adhesion properties. The use of these grids in combination with PTA for contrast in the TEM is suitable for subsequent analysis by TERS. The study reports fundamental modifications and optimizations of the negative staining EM method that allows a combination with near-field Raman spectroscopy to acquire a spectroscopic signature from nanoscale biological structures. This should facilitate a more precise diagnosis of single viral particles and other micro-organisms previously localized and visualized in the TEM. © 2014 The Society for Applied Microbiology.

  11. The development of the spatially correlated adjustment wavelet filter for atomic force microscopy data.

    Sikora, Andrzej; Rodak, Aleksander; Unold, Olgierd; Klapetek, Petr

    2016-12-01

    In this paper a novel approach for the practical utilization of the 2D wavelet filter in terms of the artifacts removal from atomic force microscopy measurements results is presented. The utilization of additional data such as summary photodiode signal map is implemented in terms of the identification of the areas requiring the data processing, filtering settings optimization and the verification of the process performance. Such an approach allows to perform the filtering parameters adjustment by average user, while the straightforward method requires an expertise in this field. The procedure was developed as the function of the Gwyddion software. The examples of filtering the phase imaging and Electrostatic Force Microscopy measurement result are presented. As the wavelet filtering feature may remove a local artifacts, its superior efficiency over similar approach with 2D Fast Fourier Transformate based filter (2D FFT) can be noticed. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Study of NaCl:Mn2+ nanostructures in the Suzuki phase by optical spectroscopy and atomic force microscopy

    Mejía-Uriarte, E.V.; Kolokoltsev, O.; Navarrete Montesinos, M.; Camarillo, E.; Hernández A, J.; Murrieta S, H.

    2015-01-01

    NaCl:Mn 2+ nanostructures in the Suzuki phase have been studied by fluorescence (emission and excitation) spectroscopy and atomic force microscopy (AFM) as a function of temperature. The “as-grown” samples give rise to two broad emission bands that peak at 508 (green emission) and 610 nm (red emission). The excitation spectrum shows peaks at 227 nm and 232 nm for emission wavelengths at 508 nm and 610 nm, respectively. When the samples are heated continuously from room temperature up to 220 °C, the green emission (associated to the excitation peak at 227 nm) disappears at a temperature close to 120 °C, whilst only the red emission remains, which is characteristic of manganese ions. AFM images on the (0 0 1) surface (freshly cleaved) show several conformations of nanostructures, such as disks of 20–50 nm in diameter. Particularly, the images also reveal nanostructures with rectangular shape of ~280×160 nm 2 and ~6 nm height; these are present only in samples with green emission associated to the Suzuki phase. Then, the evidence suggests that this topographic configuration might be related to the interaction with the first neighbors and the next neighbors, according to the configuration that has been suggested for the Suzuki phase. - Highlights: • NaCl:Mn 2+ single crystals in the Suzuki phase contain rectangular nanostructures. • Double emission of manganese ions: green (508 nm) and red (610 nm) bands. • The excitation peak at 227 nm is attributed to rectangular nanostructures. • The green emission band associated to Suzuki phase is extinguished at 120 °C

  13. Correlations of the chemical shift on fasly rotating biological solids by means of NMR spectroscopy

    Herbst, Christian

    2010-01-01

    The basic aim of the thesis was the development and improvement of homo- and heteronuclear feedback sequences for the generation of correlation spectra of the chemical shift. In a first step the possibility of the acquisition of 13 C- 13 correlation spectra of the chemical shift by means of inversion pulses with low RF power factor was studied. Furthermore it was shown that broad-band phase-modulated inversion and universal rotational pulses can be constructed by means of global optimization procedures like the genetic algorithms under regardment of the available RF field strength. By inversion, universal rotational, and 360 pulses as starting values of the optimization efficient homonuclear CN n ν and RN n ν mixing sequences as well as heteronuclear RN n ν s ,ν k feedback sequences were generated. The satisfactory power of the numerically optimized sequences was shown by means of the simulation as well by means of correlation experiments of the chemical shift of L-histidine, L-arginine, and the (CUG) 97 -RNA. This thesis deals furthermore with the possibility to acquire simultaneously different signals with several receivers. By means of numerically optimized RN n ν s ,ν k pulse sequences both 15 N- 13 C and 13 C- 15 N correlation spectra were simultaneously generated. Furthermore it could be shown that the simultaneous acquisition of 3D- 15 N- 13 C- 13 C and 13 C- 15 N-( 1 H)- 1 H correlation spectra is possible. By this in only one measurement process resonance assignments can be met and studies of the global folding performed. A further application of several receivers is the simultaneous acquisition of CHHC, NHHN, NHHC, as well as CHHN spectra. By such experiments it is possible to characterize the hydrogen-bonding pattern and the glycosidic torsion angle χ in RNA. This was demonstrated by means of the (CUG) 97 -RNA. The simultaneous acquisition of all relevant crossing signals of the correlation spectra leads not only to an essential time saving, but

  14. The development of the spatially correlated adjustment wavelet filter for atomic force microscopy data

    Sikora, Andrzej, E-mail: sikora@iel.wroc.pl [Electrotechnical Institute, Division of Electrotechnology and Materials Science, M. Skłodowskiej-Curie 55/61, 50-369 Wrocław (Poland); Rodak, Aleksander [Faculty of Electronics, Wrocław University of Technology, Janiszewskiego 11/17, 50-372 Wrocław (Poland); Unold, Olgierd [Institute of Computer Engineering, Control and Robotics, Faculty of Electronics, Wrocław University of Technology, Janiszewskiego 11/17, 50-372 Wrocław (Poland); Klapetek, Petr [Czech Metrology Institute, Okružní 31, 638 00 Brno (Czech Republic)

    2016-12-15

    In this paper a novel approach for the practical utilization of the 2D wavelet filter in terms of the artifacts removal from atomic force microscopy measurements results is presented. The utilization of additional data such as summary photodiode signal map is implemented in terms of the identification of the areas requiring the data processing, filtering settings optimization and the verification of the process performance. Such an approach allows to perform the filtering parameters adjustment by average user, while the straightforward method requires an expertise in this field. The procedure was developed as the function of the Gwyddion software. The examples of filtering the phase imaging and Electrostatic Force Microscopy measurement result are presented. As the wavelet filtering feature may remove a local artifacts, its superior efficiency over similar approach with 2D Fast Fourier Transformate based filter (2D FFT) can be noticed. - Highlights: • A novel approach to 2D wavelet-based filter for atomic force microscopy is shown. • The additional AFM measurement signal is used to adjust the filter. • Efficient removal of the local interference phenomena caused artifacts is presented.

  15. The development of the spatially correlated adjustment wavelet filter for atomic force microscopy data

    Sikora, Andrzej; Rodak, Aleksander; Unold, Olgierd; Klapetek, Petr

    2016-01-01

    In this paper a novel approach for the practical utilization of the 2D wavelet filter in terms of the artifacts removal from atomic force microscopy measurements results is presented. The utilization of additional data such as summary photodiode signal map is implemented in terms of the identification of the areas requiring the data processing, filtering settings optimization and the verification of the process performance. Such an approach allows to perform the filtering parameters adjustment by average user, while the straightforward method requires an expertise in this field. The procedure was developed as the function of the Gwyddion software. The examples of filtering the phase imaging and Electrostatic Force Microscopy measurement result are presented. As the wavelet filtering feature may remove a local artifacts, its superior efficiency over similar approach with 2D Fast Fourier Transformate based filter (2D FFT) can be noticed. - Highlights: • A novel approach to 2D wavelet-based filter for atomic force microscopy is shown. • The additional AFM measurement signal is used to adjust the filter. • Efficient removal of the local interference phenomena caused artifacts is presented.

  16. NMR spectroscopy, Hammett correlations and biological activity of some Schiff bases derived from piperonal

    Echevarria, Aurea; Giesbrecht, Astrea

    1999-01-01

    A series of eleven Schiff Bases have been synthesized. They were obtained by condensation of piperonal (3,4-methylenedioxybenzaldehyde) with the corresponding aromatic primary amines. Their 1 H and 13 C-NMR spectra have been obtained and the Hammett correlations including chemical shifts and the substituent constants (σ p , σR e σI) were studied. Linear and bilinear significant correlations were observed for iminic carbon (C-α) and C-1 ' , showing a more significant resonance effect on chemical shifts. The chemical shifts for C-4 ' were highly affected by substituent effects, especially for halogens in the expected direction. Their biological activity against microorganisms has also been measured and significant activity was showed against Epidermophyton floccosum. The biological activity did not give a reasonable relationship with electronic effects. (author)

  17. Characterisation of the membrane affinity of an isoniazide peptide conjugate by tensiometry, atomic force microscopy and sum-frequency vibrational spectroscopy, using a phospholipid Langmuir monolayer model.

    Hill, Katalin; Pénzes, Csanád Botond; Schnöller, Donát; Horváti, Kata; Bosze, Szilvia; Hudecz, Ferenc; Keszthelyi, Tamás; Kiss, Eva

    2010-10-07

    Tensiometry, sum-frequency vibrational spectroscopy, and atomic force microscopy were employed to assess the cell penetration ability of a peptide conjugate of the antituberculotic agent isoniazide. Isoniazide was conjugated to peptide (91)SEFAYGSFVRTVSLPV(106), a functional T-cell epitope of the immunodominant 16 kDa protein of Mycobacterium tuberculosis. As a simple but versatile model of the cell membrane a phospholipid Langmuir monolayer at the liquid/air interface was used. Changes induced in the structure of the phospholipid monolayer by injection of the peptide conjugate into the subphase were followed by tensiometry and sum-frequency vibrational spectroscopy. The drug penetrated lipid films were transferred to a solid support by the Langmuir-Blodgett technique, and their structures were characterized by atomic force microscopy. Peptide conjugation was found to strongly enhance the cell penetration ability of isoniazide.

  18. Gender Differences in Musculoskeletal Lipid Metabolism as Assessed by Localized Two-Dimensional Correlation Spectroscopy

    S. Sendhil Velan; Department of Exercise Physiology, West Virginia University School of Medicine, Morgantown, West Virginia, U.S.A.

    2008-01-01

    Full Text Available Gender differences in lipid metabolism are poorly understood and difficult to study using conventional approaches. Magnetic resonance spectroscopy (MRS permits non-invasive investigation of lipid metabolism. We employed novel two- dimensional MRS techniques to quantify intramyocellular (IMCL and extramyocellular (EMCL lipid compartments and their degree of unsaturation in normal weight adult male and female subjects. Using muscle creatine (Cr for normalization, a statistically significant (p 0.05 increase in IMCL/Cr (7.8 ± 1.6 and EMCL/Cr (22.5 ± 3.6 for female subjects was observed (n = 8, as compared to IMCL/Cr (5.9 ± 1.7 and EMCL/Cr (18.4 ± 2.64 for male subjects. The degree of unsaturation within IMCL and EMCL was lower in female subjects, 1.3 ± 0.075 and 1.04 ± 0.06, respectively, as compared to that observed in males (n = 8, 1.5 ± 0.08 and 1.12 ± 0.03, respectively (p 0.05 male vs female for both comparisons. We conclude that certain salient gender differences in lipid metabolism can be assessed noninvasively by advanced MRS approaches.

  19. High-spin spectroscopy of {sup 168}Yb and the reduction of pairing correlations

    Oliveira, J R.B. [Sao Paulo Univ., SP (Brazil); Stephens, F S; Deleplanque, M A; Diamond, R M [Lawrence Berkeley Lab., CA (United States); Draper, J E; Rubel, E; Duyar, C [California Univ., Davis, CA (United States); Beacker, J A; Henry, E A; Roy, N [Lawrence Livermore National Lab., CA (United States); Beausang, C W [Liverpool Univ. (United Kingdom); Frauendorf, S [Institut fur Kern und Hadronen Physik, F2-Rossendorf, Dresden (Germany)

    1992-08-01

    The high spin states of {sup 168}Yb were investigated by means of in-beam gamma spectroscopy with the High Energy Resolution Array at the 88 in. cyclotron of the Lawrence Berkeley Laboratory. The {sup 168}Yb nucleus was produced in the reaction {sup 48}Ca({sup 124}Sn,4n) at 210 MeV. Five bands previously reported were confirmed; additionally, four other bands, two extending to spins as high as 36 {Dirac_h} were observed. Cranked shell models suggest that one of the new bands can be interpreted as the continuation of the ground state band (above the AB crossing frequency) crossing into the four-quasiparticle band ABCD at about 0.38 MeV. Both relative alignment and Routhians are in good agreement with the experimental values. However, these calculations were done at constant pairing strength, which is not expected to be good at high rotational frequencies where one or more crossings have occurred in each band. Particle-hole calculations (with no pairing) were done for {sup 168}Yb as well as for other N {approx_equal} 98 nuclei for which good experimental data are available. In most cases, it is possible to associate a particle-hole configuration for each band observed at very high rotational frequencies, and the overall description is good. At frequencies below the first crossing, a full pairing calculation is necessary to describe the bands properly. At intermediate frequencies, the pairing strength is believed to be intermediate. 3 figs.

  20. Ultrafast optical pump terahertz-probe spectroscopy of strongly correlated electron materials

    Averitt, R.D.; Taylor, Antoinette J.; Thorsmolle, V.K.; Jia, Quanxi; Lobad, A.I.; Trugman, S.A.

    2001-01-01

    We have used optical-pump far-infrared probe spectroscopy to probe the low energy electron dynamics of high temperature superconductors and colossal magnetoresistance manganites. For the superconductor YBa2Cu3O7, picosecond conductivity measurements probe the interplay between Cooper-pairs and quasiparticles. In optimally doped films, the recovery time for long-range phase-coherent pairing increases from ∼1.5 ps at 4K to ∼3.5 ps near Tc, consistent with the closing of the superconducting gap. For underdoped films, the measured recovery time is temperature independent (3.5 ps) in accordance with the presence of a pseudogap. Ultrafast picosecond measurements of optically induced changes in the absolute conductivity of La0:7M0:3MnO3 thin films (M = Ca, Sr) from 10K to ∼0.9Tc reveal a two-component relaxation. A fast, ∼2 ps, conductivity decrease arises from optically induced modification of the effective phonon temperature. The slower component, related to spin-lattice relaxation, has a lifetime that increases upon approaching Tc from below in accordance with an increasing spin specific heat. Our results indicate that for T<< Tc, the conductivity is determined by incoherent phonons while spin fluctuations dominate near Tc.

  1. Gender Differences in Musculoskeletal Lipid Metabolism as Assessed by Localized Two-Dimensional Correlation Spectroscopy

    S. Sendhil Velan

    2008-01-01

    Full Text Available Gender differences in lipid metabolism are poorly understood and difficult to study using conventional approaches. Magnetic resonance spectroscopy (MRS permits non-invasive investigation of lipid metabolism. We employed novel two-dimensional MRS techniques to quantify intramyocellular (IMCL and extramyocellular (EMCL lipid compartments and their degree of unsaturation in normal weight adult male and female subjects. Using muscle creatine (Cr for normalization a statistically significant (p < 0.05 increase in IMCL/Cr (7.8 ± 1.6 and EMCL/Cr (22.5 ± 3.6 for female subjects was observed (n = 8, as compared to IMCL/Cr (5.9 ± 1.7 and EMCL/Cr (18.4 ± 2.64 for male subjects. The degree of unsaturation within IMCL and EMCL was lower in female subjects, 1.3 ± 0.075 and 1.04 ± 0.06, respectively, as compared to that observed in males (n = 8, 1.5 ± 0.08 and 1.12 ± 0.03, respectively (p < 0.05 male vs female for both comparisons. We conclude that certain salient gender differences in lipid metabolism can be assessed noninvasively by advanced MRS approaches.

  2. High-resolution high-speed dynamic mechanical spectroscopy of cells and other soft materials with the help of atomic force microscopy.

    Dokukin, M; Sokolov, I

    2015-07-28

    Dynamic mechanical spectroscopy (DMS), which allows measuring frequency-dependent viscoelastic properties, is important to study soft materials, tissues, biomaterials, polymers. However, the existing DMS techniques (nanoindentation) have limited resolution when used on soft materials, preventing them from being used to study mechanics at the nanoscale. The nanoindenters are not capable of measuring cells, nanointerfaces of composite materials. Here we present a highly accurate DMS modality, which is a combination of three different methods: quantitative nanoindentation (nanoDMA), gentle force and fast response of atomic force microscopy (AFM), and Fourier transform (FT) spectroscopy. This new spectroscopy (which we suggest to call FT-nanoDMA) is fast and sensitive enough to allow DMS imaging of nanointerfaces, single cells, while attaining about 100x improvements on polymers in both spatial (to 10-70 nm) and temporal resolution (to 0.7 s/pixel) compared to the current art. Multiple frequencies are measured simultaneously. The use of 10 frequencies are demonstrated here (up to 300 Hz which is a rather relevant range for biological materials and polymers, in both ambient conditions and liquid). The method is quantitatively verified on known polymers and demonstrated on cells and polymers blends. Analysis shows that FT-nanoDMA is highly quantitative. The FT-nanoDMA spectroscopy can easily be implemented in the existing AFMs.

  3. Investigation of electric quadrupole interaction in TiO2 by means of perturbed gamma-gamma angular correlation spectroscopy

    Martucci, Thiago; Ramos, Juliana Marques; Carbonari, Artur Wilson; Silva, Andreia S.; Saxena, Rajendra Narain

    2011-01-01

    TiO 2 has called attention in both basic research and technological applications as an energy converter in solar cells, photo catalyst for water purification, sunscreen material, drug detection, and other applications. In addition TiO 2 is a candidate for use in medical devices, food preparation surfaces, air conditioning filters and sanitary ware surfaces.TiO 2 has two crystalline phases: anatase and rutile. The structural properties and hyperfine interactions in TiO 2 were investigated by perturbed gamma-gamma angular correlation (PAC) spectroscopy using 111 In and 181 Hf as probe nuclei. The PAC spectroscopy provides information on crystalline and electronic structure at an atomic scale. In the present work, PAC measurements on TiO 2 were focused on the development of a methodology to prepare bulk samples, which have been characterized by conventional techniques such as x-ray diffraction, (XRD), scanning electron microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). The samples were prepared by the sol gel Pechini method. The resulting powders were annealed at different temperatures in a tubular furnace under nitrogen atmosphere. The PAC measurements were carried out at room temperature in air. The occupation fraction of the probe nuclei reached 50% when the sample was annealed at 1373K and after measured at room temperature.In this case the frequency measured in site 1 is in agreement with that found in literature for substitutional titanium site in rutile structure when 111 In were used as probe nuclei. It was measured a frequency more closely to that was found in literature[7] for the case in which 181 Hf were used as probe nuclei when the sample annealed at 1373 K and measured at 973 K. (author)

  4. Real-time autocorrelator for fluorescence correlation spectroscopy based on graphical-processor-unit architecture: method, implementation, and comparative studies

    Laracuente, Nicholas; Grossman, Carl

    2013-03-01

    We developed an algorithm and software to calculate autocorrelation functions from real-time photon-counting data using the fast, parallel capabilities of graphical processor units (GPUs). Recent developments in hardware and software have allowed for general purpose computing with inexpensive GPU hardware. These devices are more suited for emulating hardware autocorrelators than traditional CPU-based software applications by emphasizing parallel throughput over sequential speed. Incoming data are binned in a standard multi-tau scheme with configurable points-per-bin size and are mapped into a GPU memory pattern to reduce time-expensive memory access. Applications include dynamic light scattering (DLS) and fluorescence correlation spectroscopy (FCS) experiments. We ran the software on a 64-core graphics pci card in a 3.2 GHz Intel i5 CPU based computer running Linux. FCS measurements were made on Alexa-546 and Texas Red dyes in a standard buffer (PBS). Software correlations were compared to hardware correlator measurements on the same signals. Supported by HHMI and Swarthmore College

  5. The Correlation of Radon Concentration with Various Building Attributes at U.S. Air Force Bases

    1992-08-01

    these daughter products that continue to decay giving off radiation which can then lead to the development of lung cancer . The United States Air Force...USAF) is concerned about the increased risk of developing lung cancer by persons exposed to elevated levels of radon in their domiciles and in their...CONOM 0 S * 0 0 a N&NO.)C Nowfum - a w em C mec0- C Mama - 00 4.4 .Q0 0 40 VO O 02ýCt > a MW 0 5 O 0 500 0 ’-ONM > Co S- -W N 00l 0 N 0 O--0 CPe go - Na Wm

  6. Condom Use Behaviors and Correlates of Condom Use in the Botswana Defence Force

    2013-10-10

    may engage in risky sexual behaviours, such as having unprotected sex with sex workers .4 Furthermore, studies have found high HIV risk- taking...did not use a condom the last time they had sex with a sex worker . 11 And in the 1Naval Health Research Center, Department of Defense HIV /AIDS...only found in 16-20% of participants,7 and 41% did not use a condom the last time they had sex with a sex worker .4 In the Rwanda Defence Forces, 24

  7. Proton and O sup(2-) ion diffusion studied by γ-γ angular correlation and by impedance spectroscopy

    Oliveira Damasceno, O. de.

    1988-01-01

    The interaction of sup(181) Ta quadrupole momentum with electric field gradient was measured by perturbed angular correlation spectroscopy in polycrystalline samples of hafnium hydride and hafnium oxide. The measurements were done as function of the temperature, in cubic and tetragonal phases of hafnium hydride. In the cubic phase, spin relaxation effects related to proton diffusion with activation energy of 0.43 ± 0.05eV were observed, and in the tetragonal phase, the interaction was purely static. The hafnium oxide was studied in cubic phase stabilized by addition of calcium oxide or magnesium oxide. Relaxation effects probably due to O sup(2-) ion diffusion were verified. The protonic conduction in K H sub(2) PO sub(4) was studied by impedance spectroscopy, in pellets prepared by melting or powder compression using silver and platinum electrodes. In the case of silver electrodes, in the low frequency region, the impedance spectrum tends to a 45 sup(0) typical line of the diffusion process. It was attributed to hydrogen injection in the electrode. Heating k H sub(2) PO sub(4) at about 230 sup(0)C modifies significantly the electric properties. Two relaxation annealing reduced conductivities to constant values independently of preparation method. (author)

  8. Nuclear spectroscopy study of the 117 Sn by the angular correlation technique

    Borges, Joao Baptista

    1977-01-01

    The directional correlation of gamma cascade (553-159) keV populated in 117 Sn through the β - decay of 117 In has been measured. An automatic gamma spectrometer utilizing Ge(Li) and NaI (Tl) detectors was used to measure the angular correlation. The results are analysed in terms of the multipole mixing ratio for the 159 keV transition in 117 Sn. The results are: A 22 = -0 064±0.005, A 44 = 0.005±0.007 with δ(E2/M1) 159keV = 0.036+0.021. The life time of the 159 keV state has also been determined by using the plastic scintillator detectors, and utilizing the delayed gamma-gamma coincidence method the resulting value of the life time is T 1/2 = 275±15 psec. Further measurements have been carried out to determine the nuclear g-factor of the 159 keV state utilizing the NaI(Tl) detectors and an external magnetic field of 25.5 k Gauss. The method of 'integral rotation with reverse field and constant angle' was utilized for the determination of the g-factor with the resulting value of g(159 keV) = +0.47±0.10. The experimental results are discussed in terms of single particle model and the pairing plus quadrupole model of Kisslinger and Sorensen. (author)

  9. Epileptic rat brain tissue analyzed by 2D correlation Raman spectroscopy

    Sacharz, Julia; Wesełucha-Birczyńska, Aleksandra; Zięba-Palus, Janina; Lewandowski, Marian H.; Kowalski, Rafał; Palus, Katarzyna; Chrobok, Łukasz; Moskal, Paulina; Birczyńska, Malwina; Sozańska, Agnieszka

    2018-01-01

    Absence epilepsy is the neurological disorder characterized by the pathological spike-and wave discharges present in the electroencephalogram, accompanying a sudden loss of consciousness. Experiments were performed on brain slices obtained from young male WAG/Rij rats (2-3 weeks old), so that they were sampled before the appearance of brain-damaging seizures symptoms. Two differing brain areas of the rats' brain tissue were studied: the somatosensory cortex (Sc) and the dorsal lateral geniculate nucleus of the thalamus (DLG). The Raman spectra of the fresh brain scraps, kept during measurements in artificial cerebrospinal fluid, were collected using as an excitation source 442 nm, 514.5 nm, 785 nm and 1064 nm laser line. The average spectra were analyzed by 2D correlation method regarding laser line as an external perturbation. In 2D synchronous spectra positive auto-peaks corresponding to the Cdbnd C stretching and amide I band vibrations show maxima at 1660 cm- 1 and 1662 cm- 1 for Sc and DLG, respectively. The prominent auto-peak at 2937 cm- 1, originated from the CH3 mode in DLG brain area, seems to indicate the importance of methylation, considered to be significant in epileptogenesis. Synchronous and asynchronous correlations peaks, glutamic acid and gamma-aminobutyric acid (GABA), appear in Sc and DLG, respectively. In the 1730-1600 cm- 1 range occur cross-peaks which appearance might be triggered by glial fibrillary acidic protein (GFAP) activation.

  10. Studies of metal binding by the iron transport protein transferrin using time differential perturbed angular correlation spectroscopy

    Then, G.M.

    1987-01-01

    The binding of the transition metal hafnium to transferrin was studied under various chemical conditions using time differential perturbed γγ angular correlation spectroscopy (TDPAC). Observing the electric quadrupole interaction of the 181 Hf probe nuclei size and symmetry of the electric field gradient induced by the ligands of the metal ions can be determined. The experimental data suggest how homogeneous the binding conditions are and to which extend relaxation phenomena are involved. Due to the excellent time resolution obtained with new BaF 2 detectors the quadrupole coupling parameters of 181 Hf-transferrin could be determined very accurately. Under nearly physiological conditions different binding configurations were quantitatively characterized by spectroscopic means and distinguished with high specificity. (orig./PW) [de

  11. Fibrillation mechanism of a model intrinsically disordered protein revealed by 2D correlation deep UV resonance Raman spectroscopy.

    Sikirzhytski, Vitali; Topilina, Natalya I; Takor, Gaius A; Higashiya, Seiichiro; Welch, John T; Uversky, Vladimir N; Lednev, Igor K

    2012-05-14

    Understanding of numerous biological functions of intrinsically disordered proteins (IDPs) is of significant interest to modern life science research. A large variety of serious debilitating diseases are associated with the malfunction of IDPs including neurodegenerative disorders and systemic amyloidosis. Here we report on the molecular mechanism of amyloid fibrillation of a model IDP (YE8) using 2D correlation deep UV resonance Raman spectroscopy. YE8 is a genetically engineered polypeptide, which is completely unordered at neutral pH yet exhibits all properties of a fibrillogenic protein at low pH. The very first step of the fibrillation process involves structural rearrangements of YE8 at the global structure level without the detectable appearance of secondary structural elements. The formation of β-sheet species follows the global structural changes and proceeds via the simultaneous formation of turns and β-strands. The kinetic mechanism revealed is an important new contribution to understanding of the general fibrillation mechanism proposed for IDP.

  12. X-ray Photon Correlation Spectroscopy Study on Dynamics of the Free Surface in Entangled Polystyrene Melt Films

    Koga, Tadanori; Li Chunhua; Endoh, Maya K; Narayanan, Suresh; Lurio, Laurence; Sinha, Sunil K

    2011-01-01

    The dynamics of polymer chains near the surface of a melt and within thin films remains a subject of inquiry along with the nature of the glass transition in these systems. Recent studies show that the properties of the free surface region are crucial in determining the anomalous glass transition temperature (T g ) reduction of polymer thin films. In this study, by embedding 'dilute' gold nanoparticles in polystyrene (PS) thin films as 'markers', we could successfully probe the diffusive Brownian motion which tracks the local viscosity both at the free surface and within the rest of the single PS thin film far above bulk T g . The technique used was X-ray photon correlation spectroscopy with resonance-enhanced X-rays that allows us to independently measure the motion in the regions of interest at the nanometer scale. We found the presence of the surface reduced viscosity layer in entangled PS thin films at T>>T g .

  13. Nano-viscosity of supercooled liquid measured by fluorescence correlation spectroscopy: Pressure and temperature dependence and the density scaling

    Meier, G.; Gapinski, J.; Ratajczyk, M.; Lettinga, M. P.; Hirtz, K.; Banachowicz, E.; Patkowski, A.

    2018-03-01

    The Stokes-Einstein relation allows us to calculate apparent viscosity experienced by tracers in complex media on the basis of measured self-diffusion coefficients. Such defined nano-viscosity values can be obtained through single particle techniques, like fluorescence correlation spectroscopy (FCS) and particle tracking (PT). In order to perform such measurements, as functions of pressure and temperature, a new sample cell was designed and is described in this work. We show that this cell in combination with a long working distance objective of the confocal microscope can be used for successful FCS, PT, and confocal imaging experiments in broad pressure (0.1-100 MPa) and temperature ranges. The temperature and pressure dependent nano-viscosity of a van der Waals liquid obtained from the translational diffusion coefficient measured in this cell by means of FCS obeys the same scaling as the rotational relaxation and macro-viscosity of the system.

  14. Heat transfer experiments and correlations for natural and forced circulations of water in rod bundles at low Reynolds numbers

    Kim, Sung-Ho; El-Genk, Mohamed S.; Rubio, Reuben A.; Bryson, James W.; Foushee, Fabian C.

    1988-01-01

    Experimental heat transfer studies were conducted for fully developed forced and natural flows of water through seven uniformly heated rod bundles, triangularly arrayed with P/D = 1.25, 1.38, and 1.5. In forced circulation experiments, Re ranged from 80 to 50,000 and Pr from 3 to 8.5, while in natural circulation, Re varied from 260 to 2,000, and Ra q from 8 x 10 8 to 2.5 x 10 8 . The forced flow data fell into the two basic flow regimes: turbulent and laminar flow. At the transition between these regimes, Re, which varied from 2,200 for P/D = 1.25 to 5,500 for P/D = 1.5, increased linearly with P/D. The heat transfer data for turbulent flow was within ±15 percent of Weisman's correlation, which was developed for fully developed turbulent flow in rod bundles at Re > 25,000. The laminar flow data showed the dependence of Nu on Re to be weaker than that for turbulent flow, but the exponent of Re increased with P/D: Nu = A Re B Pr 1/3 , where A is equal to 1.061, 0.511, and 0.346 for P/D = 1.25, 1.38 and 1.5, respectively, and B is a linear function of P/D (B = 0.797 P/D - 0.656). Natural circulation data indicated that rod spacing only slightly affected heat transfer, and Nu increased proportionally to Ra 0.25 ; Nu = 0.272 Ra q 0.25 . The application of the results to SNL's ACRR indicated that although the core is cooled by natural convection, either the natural circulation correlation or the forced turbulent flow correlation can be used to accurately predict the single phase heat transfer coefficient in the ACRR. These results were concluded because of the high Rayleigh and Reynolds numbers in the ACRR. The ACRR operates near the boundary between mixed and forced turbulent flow regimes: consequently, achieving the high heat transfer coefficient was possible with natural circulation. (author)

  15. Simultaneous membrane interaction of amphipathic peptide monomers, self-aggregates and cargo complexes detected by fluorescence correlation spectroscopy.

    Vasconcelos, Luís; Lehto, Tõnis; Madani, Fatemeh; Radoi, Vlad; Hällbrink, Mattias; Vukojević, Vladana; Langel, Ülo

    2018-02-01

    Peptides able to translocate cell membranes while carrying macromolecular cargo, as cell-penetrating peptides (CPPs), can contribute to the field of drug delivery by enabling the transport of otherwise membrane impermeable molecules. Formation of non-covalent complexes between amphipathic peptides and oligonucleotides is driven by electrostatic and hydrophobic interactions. Here we investigate and quantify the coexistence of distinct molecular species in multiple equilibria, namely peptide monomer, peptide self-aggregates and peptide/oligonucleotide complexes. As a model for the complexes, we used a stearylated peptide from the PepFect family, PF14 and siRNA. PF14 has a cationic part and a lipid part, resembling some characteristics of cationic lipids. Fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS) were used to detect distinct molecular entities in solution and at the plasma membrane of live cells. For that, we labeled the peptide with carboxyrhodamine 6G and the siRNA with Cyanine 5. We were able to detect fluorescent entities with diffusional properties characteristic of the peptide monomer as well as of peptide aggregates and peptide/oligonucleotide complexes. Strategies to avoid peptide adsorption to solid surfaces and self-aggregation were developed and allowed successful FCS measurements in solution and at the plasma membrane. The ratio between the detected molecular species was found to vary with pH, peptide concentration and the proximity to the plasma membrane. The present results suggest that the diverse cellular uptake mechanisms, often reported for amphipathic CPPs, might result from the synergistic effect of peptide monomers, self-aggregates and cargo complexes, distributed unevenly at the plasma membrane. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. In vivo 1D and 2D correlation MR spectroscopy of the soleus muscle at 7T

    Ramadan, Saadallah; Ratai, Eva-Maria; Wald, Lawrence L.; Mountford, Carolyn E.

    2010-05-01

    AimThis study aims to (1) undertake and analyse 1D and 2D MR correlation spectroscopy from human soleus muscle in vivo at 7T, and (2) determine T1 and T2 relaxation time constants at 7T field strength due to their importance in sequence design and spectral quantitation. MethodSix healthy, male volunteers were consented and scanned on a 7T whole-body scanner (Siemens AG, Erlangen, Germany). Experiments were undertaken using a 28 cm diameter detunable birdcage coil for signal excitation and an 8.5 cm diameter surface coil for signal reception. The relaxation time constants, T1 and T2 were recorded using a STEAM sequence, using the 'progressive saturation' method for the T1 and multiple echo times for T2. The 2D L-Correlated SpectroscopY (L-COSY) method was employed with 64 increments (0.4 ms increment size) and eight averages per scan, with a total time of 17 min. ResultsT1 and T2 values for the metabolites of interest were determined. The L-COSY spectra obtained from the soleus muscle provided information on lipid content and chemical structure not available, in vivo, at lower field strengths. All molecular fragments within multiple lipid compartments were chemically shifted by 0.20-0.26 ppm at this field strength. 1D and 2D L-COSY spectra were assigned and proton connectivities were confirmed with the 2D method. ConclusionIn vivo 1D and 2D spectroscopic examination of muscle can be successfully recorded at 7T and is now available to assess lipid alterations as well as other metabolites present with disease. T1 and T2 values were also determined in soleus muscle of male healthy volunteers.

  17. Transport and accumulation of PVP-Hypericin in cancer and normal cells characterized by image correlation spectroscopy techniques.

    Penjweini, Rozhin; Smisdom, Nick; Deville, Sarah; Ameloot, Marcel

    2014-05-01

    PVP-Hypericin (PVP: polyvinylpyrrolidone) is a potent anti-cancer photosensitizer for photodynamic diagnosis (PDD) and therapy (PDT). However, cellular targets and mechanisms involved in the cancer-selectivity of the photosensitizer are not yet fully understood. This paper gives new insights into the differential transport and localization of PVP-Hypericin in cancer and normal cells which are essential to unravel the mechanisms of action and cancer-selectivity. Temporal (TICS) and spatiotemporal (STICS) image correlation spectroscopy are used for the assessment of PVP-Hypericin diffusion and/or velocity in the case of concerted flow in human cervical epithelial HeLa and human lung carcinoma A549 cells, as well as in human primary dendritic cells (DC) and human peripheral blood mononuclear cells (PBMC). Spatiotemporal image cross-correlation spectroscopy (STICCS) based on organelle specific fluorescent labeling is employed to study the accumulation of the photosensitizer in nucleus, mitochondria, early-endosomes and lysosomes of the cells and to assess the dynamics of co-migrating molecules. Whereas STICS and TICS did not show a remarkable difference between the dynamics of PVP-Hypericin in HeLa, A549 and DC cells, a significantly different diffusion rate of the photosensitizer was measured in PBMC. STICCS detected a stationary accumulation of PVP-Hypericin within the nucleus, mitochondria, early endosomes and lysosomes of HeLa and A549 cells. However, significant flow due to the directed motion of the organelles was detected. In contrast, no accumulation in the nucleus and mitochondria of DC and PBMC could be monitored. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Prevalence of sexual victimization and correlates of forced sex in Japanese men who have sex with men.

    Yasuharu Hidaka

    Full Text Available Studies of men who have sex with men (MSM in diverse geographic and cultural contexts have identified health challenges affecting this population. MSM might be particularly vulnerable to sexual victimization and forced sex. The aim of this research study was to examine prevalence of sexual victimization and correlates of forced sex among Japanese MSM. We recruited a sample of 5,731 Japanese MSM who completed an internet-administered survey. Participants reported on history of different types of sexual victimization, unprotected anal sex, other health risk behaviors, exposure to gay-related teasing and bullying, depression, and suicidality. Over one-fifth of the sample (21.4% reported experiencing at least one form of sexual victimization, and 8.7% reported a history of forced sex. MSM who had ever experienced forced sex were significantly more likely to report experiencing psychological risks (depression OR = 1.55, 95% CI = 1.28-1.89; attempted suicide OR = 2.25, 95% CI = 1.81-2.81; other forms of bullying OR = 1.38, 95% CI = 1.13-1.68 and other behavioral risks (unprotected anal sex OR = 1.56, 95% CI = 1.29-1.90; sex venue attendance OR = 1.27, 95% CI = 1.04-1.54; methamphetamine use OR = 1.57, 95% CI  = 1.05-1.36, compared to MSM who had not experienced forced sex. Efforts to develop holistic and integrated health services for Japanese MSM are warranted, particularly related to psychosocial determinants of HIV prevention. However, due to cultural factors that emphasize familial and social relations and that stigmatize same-sex behavior, Japanese MSM might experience challenges to seeking social support and health services. Interventions must be provided in safe and non-judgmental settings where Japanese MSM feel comfortable disclosing their health and social support needs.

  19. Correlation between near-infrared spectroscopy and magnetic resonance imaging of rat brain oxygenation modulation

    Chen Yu; Tailor, Dharmesh R; Intes, Xavier; Chance, Britton

    2003-01-01

    We measure the tissue oxygen and haemoglobin concentrations in the rat brain during modulation of inhaled oxygen concentration (FiO 2 ), using non-invasive frequency domain near-infrared oximetry. The rise in oxygenated haemoglobin concentration and the decline in deoxygenated haemoglobin concentration are demonstrated in correspondence with the modulation of FiO 2 , which is changed from 20% to 100% in increments of 20%. Furthermore, the tissue oxygenation saturation also shows the corresponding trend and changes ranging from approximately 70% to 90%. The relative changes in deoxygenated haemoglobin concentration are compared to the blood-oxygenation-level-dependent (BOLD) MRI signal recorded during a similar FiO 2 protocol. A linear relationship with high correlation coefficient between the relative changes in the BOLD MRI signal and the NIRS signal is observed

  20. Correlation between near-infrared spectroscopy and magnetic resonance imaging of rat brain oxygenation modulation

    Chen Yu [Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA (United States); Tailor, Dharmesh R [Department of Bioengineering, University of Pennsylvania, Philadelphia, PA (United States); Intes, Xavier [Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA (United States); Chance, Britton [Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA (United States)

    2003-02-21

    We measure the tissue oxygen and haemoglobin concentrations in the rat brain during modulation of inhaled oxygen concentration (FiO{sub 2}), using non-invasive frequency domain near-infrared oximetry. The rise in oxygenated haemoglobin concentration and the decline in deoxygenated haemoglobin concentration are demonstrated in correspondence with the modulation of FiO{sub 2}, which is changed from 20% to 100% in increments of 20%. Furthermore, the tissue oxygenation saturation also shows the corresponding trend and changes ranging from approximately 70% to 90%. The relative changes in deoxygenated haemoglobin concentration are compared to the blood-oxygenation-level-dependent (BOLD) MRI signal recorded during a similar FiO{sub 2} protocol. A linear relationship with high correlation coefficient between the relative changes in the BOLD MRI signal and the NIRS signal is observed.

  1. Microrheology of polymeric solutions using x-ray photon correlation spectroscopy

    Papagiannopoulos, A; Waigh, T A; Fluerasu, A; Fernyhough, C; Madsen, A

    2005-01-01

    We demonstrate the technique of XPCS microrheology on opaque polymeric solutions (1-20% w/w) using colloidal silica probes. The short time decay of the intensity correlation function provides the mean square displacement (MSD) of the colloidal probes. The MSDs of the probes are subsequently transformed using the generalized Stokes-Einstein equation, allowing the linear viscoelastic spectra of a biopolymer (gellan) and a synthetic polyelectrolyte (polystyrene sulfonate, PSS) to be calculated over two decades of frequency. MSDs can be measured that are two orders of magnitude smaller than those possible with video particle tracking microrheology, with a sensitivity of ∼10 nm s -1 for displacements of ∼nms. The XPCS data for water, glycerol and PSS combs are in agreement with video particle tracking microrheology experiments performed at lower polymer concentrations. (letter to the editor)

  2. A systematic study of the octupole correlations in the lanthanides with realistic forces

    Egido, J.L.; Robledo, L.M.

    1992-01-01

    We have performed a systematic study of the octupole degree of freedom in the nuclei 140 Ba, 142-150 deg Ce, 144-152 Nd and 146-154 Sm. The static properties (ground state deformations, energy gaps, dipole moments, etc.) have been analyzed within the Hartree-Fock plus BCS approximation (HFBCS); for the dynamical ones (energy splittings, transition probabilities, etc.) the adiabatic time-dependent Hartree-Fock plus zero point energy in the cranking approximation (ATDHF+ZPE) has been applied. In both approximations the realistic density-dependent Gogny force has been used. In our parameter-free calculations we are able to describe very well the whole experimental systematic of energy splittings and B(E1), among others. The flatness of the whole experimental systematic of energy splittings and B(E1), among others. The flatness of the potential energy of some nuclei makes the mean field approach unreliable for such nuclei. (orig.)

  3. Use of atomic force microscopy and transmission electron microscopy for correlative studies of bacterial capsules.

    Stukalov, Oleg; Korenevsky, Anton; Beveridge, Terry J; Dutcher, John R

    2008-09-01

    Bacteria can possess an outermost assembly of polysaccharide molecules, a capsule, which is attached to their cell wall. We have used two complementary, high-resolution microscopy techniques, atomic force microscopy (AFM) and transmission electron microscopy (TEM), to study bacterial capsules of four different gram-negative bacterial strains: Escherichia coli K30, Pseudomonas aeruginosa FRD1, Shewanella oneidensis MR-4, and Geobacter sulfurreducens PCA. TEM analysis of bacterial cells using different preparative techniques (whole-cell mounts, conventional embeddings, and freeze-substitution) revealed capsules for some but not all of the strains. In contrast, the use of AFM allowed the unambiguous identification of the presence of capsules on all strains used in the present study, including those that were shown by TEM to be not encapsulated. In addition, the use of AFM phase imaging allowed the visualization of the bacterial cell within the capsule, with a depth sensitivity that decreased with increasing tapping frequency.

  4. Spatial Temporal Image Correlation Spectroscopy (STICS) for Flow Analysis with Application for Blood Flow Mapping

    Rossow, Molly; Gratton, Enrico; Mantulin, William M.

    2009-01-01

    It is important for surgeons to be able to measure blood flow in exposed arterioles during surgery. We report our progress in the development of an optical technique that will measure blood flow in surgically exposed blood vessels and enable previously difficult measurements. By monitoring optical fluctuations, the optical technique, based on Spatial Temporal Image Correlation (STICS), will directly measure the velocity of micron-scale particles--such as red blood cells. It will complement existing technology and provide qualitative measurements that were not previously possible. It relies on the concept that blood, when viewed on a small enough scale, is an inhomogeneous substance. Individual blood cells passing between a near-infrared light source and a detector will cause fluctuations in the transmitted optical signal. The speed, direction, and flow pattern of blood cells can be determined from these optical fluctuations. We present a series of computer simulations and experiments on phantom and animal systems to test this technique's ability to map complex flow patterns.

  5. Discrimination of Chinese Sauce liquor using FT-IR and two-dimensional correlation IR spectroscopy

    Sun, Su-Qin; Li, Chang-Wen; Wei, Ji-Ping; Zhou, Qun; Noda, Isao

    2006-11-01

    We applied the three-step IR macro-fingerprint identification method to obtain the IR characteristic fingerprints of so-called Chinese Sauce liquor (Moutai liquor and Kinsly liquor) and a counterfeit Moutai. These fingerprints can be used for the identification and discrimination of similar liquor products. The comparison of their conventional IR spectra, as the first step of identification, shows that the primary difference in Sauce liquor is the intensity of characteristic peaks at 1592 and 1225 cm -1. The comparison of the second derivative IR spectra, as the second step of identification, shows that the characteristic absorption in 1400-1800 cm -1 is substantially different. The comparison of 2D-IR correlation spectra, as the third and final step of identification, can discriminate the liquors from another direction. Furthermore, the method was successfully applied to the discrimination of a counterfeit Moutai from the genuine Sauce liquor. The success of the three-step IR macro-fingerprint identification to provide a rapid and effective method for the identification of Chinese liquor suggests the potential extension of this technique to the identification and discrimination of other wine and spirits, as well.

  6. Spatial Temporal Image Correlation Spectroscopy (STICS) for Flow Analysis with Application for Blood Flow Mapping (abstract)

    Rossow, Molly; Mantulin, William M.; Gratton, Enrico

    2009-04-01

    It is important for surgeons to be able to measure blood flow in exposed arterioles during surgery. We report our progress in the development of an optical technique that will measure blood flow in surgically exposed blood vessels and enable previously difficult measurements. By monitoring optical fluctuations, the optical technique, based on Spatial Temporal Image Correlation (STICS), will directly measure the velocity of micron-scale particles-such as red blood cells. It will complement existing technology and provide qualitative measurements that were not previously possible. It relies on the concept that blood, when viewed on a small enough scale, is an inhomogeneous substance. Individual blood cells passing between a near-infrared light source and a detector will cause fluctuations in the transmitted optical signal. The speed, direction, and flow pattern of blood cells can be determined from these optical fluctuations. We present a series of computer simulations and experiments on phantom and animal systems to test this technique's ability to map complex flow patterns.

  7. Reduced α-stable dynamics for multiple time scale systems forced with correlated additive and multiplicative Gaussian white noise

    Thompson, William F.; Kuske, Rachel A.; Monahan, Adam H.

    2017-11-01

    Stochastic averaging problems with Gaussian forcing have been the subject of numerous studies, but far less attention has been paid to problems with infinite-variance stochastic forcing, such as an α-stable noise process. It has been shown that simple linear systems driven by correlated additive and multiplicative (CAM) Gaussian noise, which emerge in the context of reduced atmosphere and ocean dynamics, have infinite variance in certain parameter regimes. In this study, we consider the stochastic averaging of systems where a linear CAM noise process in the infinite variance parameter regime drives a comparatively slow process. We use (semi)-analytical approximations combined with numerical illustrations to compare the averaged process to one that is forced by a white α-stable process, demonstrating consistent properties in the case of large time-scale separation. We identify the conditions required for the fast linear CAM process to have such an influence in driving a slower process and then derive an (effectively) equivalent fast, infinite-variance process for which an existing stochastic averaging approximation is readily applied. The results are illustrated using numerical simulations of a set of example systems.

  8. Correlated topographic and spectroscopic imaging by combined atomic force microscopy and optical microscopy

    Hu Dehong; Micic, Miodrag; Klymyshyn, Nicholas; Suh, Y.D.; Lu, H.P.

    2004-01-01

    Near-field scanning microscopy is a powerful approach to obtain topographic and spectroscopic characterization simultaneously for imaging biological and nanoscale systems. To achieve optical imaging at high spatial resolution beyond the diffraction limit, aperture-less metallic scanning tips have been utilized to enhance the laser illumination local electromagnetic field at the apex of the scanning tips. In this paper, we discuss and review our work on combined fluorescence imaging with AFM-metallic tip enhancement, finite element method simulation of the tip enhancement, and their applications on AFM-tip enhanced fluorescence lifetime imaging (AFM-FLIM) and correlated AFM and FLIM imaging of the living cells

  9. Vibrational correlation between conjugated carbonyl and diazo modes studied by single- and dual-frequency two-dimensional infrared spectroscopy

    Maekawa, Hiroaki; Sul, Soohwan; Ge, Nien-Hui

    2013-08-01

    We have applied infrared three-pulse photon echo and single- and dual-frequency 2D IR spectroscopy to the ester Cdbnd O and diazo Ndbnd N stretching modes in ethyl diazoacetate (EDA), and investigated their vibrational frequency fluctuations and correlation. The two modes exhibit different vibrational dynamics and 2D lineshape, which are well simulated by frequency-frequency correlation functions (FFCFs) with two decaying components. Although the FT IR spectrum shows a single Cdbnd O band, absolute magnitude 2D IR nonrephasing spectrum displays spectral signatures supporting the presence of cis and trans conformations. The cross-peak inclined toward the anti-diagonal in the dual-frequency 2D IR spectrum, indicating that the frequency fluctuations of the two modes are anticorrelated. This behavior is attributed to anticorrelated change in the bond orders when solvent and structural fluctuations causes EDA to adopt a different mixture of the two dominant resonance structures. The effects of cross FFCF on the cross-peak line shape are discussed.

  10. Correlation between memory, proton magnetic resonance spectroscopy, and interictal epileptiform discharges in temporal lobe epilepsy related to mesial temporal sclerosis.

    Mantoan, Marcele Araújo Silva; Caboclo, Luís Otávio Sales Ferreira; de Figueiredo Ferreira Guilhoto, Laura Maria; Lin, Katia; da Silva Noffs, Maria Helena; de Souza Silva Tudesco, Ivanda; Belzunces, Erich; Carrete, Henrique; Bussoletti, Renato Tavares; Centeno, Ricardo Silva; Sakamoto, Américo Ceiki; Yacubian, Elza Márcia Targas

    2009-11-01

    The aim of the study described here was to examine the relationship between memory function, proton magnetic resonance spectroscopy ((1)H-MRS) abnormalities, and interictal epileptiform discharge (IED) lateralization in patients with temporal lobe epilepsy (TLE) related to unilateral mesial temporal sclerosis. We assessed performance on tests of memory function and intelligence quotient (IQ) in 29 right-handed outpatients and 24 controls. IEDs were assessed on 30-minute-awake and 30-minute-sleep EEG samples. Patients had (1)H-MRS at 1.5 T. There was a negative correlation between IQ (P=0.031) and Rey Auditory Verbal Learning Test results (P=0.022) and epilepsy duration; between(1)H-MRS findings and epilepsy duration (P=0.027); and between N-acetylaspartate (NAA) levels and IEDs (P=0.006) in contralateral mesial temporal structures in the left MTS group. (1)H-MRS findings, IEDs, and verbal function were correlated. These findings suggest that IEDs and NAA/(Cho+Cr) ratios reflecting neural metabolism are closely related to verbal memory function in mesial temporal sclerosis. Higher interictal activity on the EEG was associated with a decline in total NAA in contralateral mesial temporal structures.

  11. In situ fluorescence spectroscopy correlates ionomer degradation to reactive oxygen species generation in an operating fuel cell.

    Prabhakaran, Venkateshkumar; Arges, Christopher G; Ramani, Vijay

    2013-11-21

    The rate of generation of reactive oxygen species (ROS) within the polymer electrolyte membrane (PEM) of an operating proton exchange member fuel cell (PEMFC) was monitored using in situ fluorescence spectroscopy. A modified barrier layer was introduced between the PEM and the electrocatalyst layer to eliminate metal-dye interactions and fluorescence resonance energy transfer (FRET) effects during measurements. Standard fuel cell operating parameters (temperature, relative humidity, and electrode potential) were systematically varied to evaluate their influence on the rate of ROS generation during PEMFC operation. Independently, the macroscopic rate of PEM degradation was measured by monitoring the fluoride ion emission rate (FER) in the effluent stream at each operating condition. The ROS generation reaction rate constant (estimated from the in situ fluorescence experiments) correlated perfectly with the measured FER across all conditions, demonstrating unequivocally for the first time that a direct correlation exists between in situ ROS generation and PEM macroscopic degradation. The activation energy for ROS generation within the PEM was estimated to be 12.5 kJ mol(-1).

  12. A Nth-order linear algorithm for extracting diffuse correlation spectroscopy blood flow indices in heterogeneous tissues.

    Shang, Yu; Yu, Guoqiang

    2014-09-29

    Conventional semi-infinite analytical solutions of correlation diffusion equation may lead to errors when calculating blood flow index (BFI) from diffuse correlation spectroscopy (DCS) measurements in tissues with irregular geometries. Very recently, we created an algorithm integrating a N th-order linear model of autocorrelation function with the Monte Carlo simulation of photon migrations in homogenous tissues with arbitrary geometries for extraction of BFI (i.e., αD B ). The purpose of this study is to extend the capability of the N th-order linear algorithm for extracting BFI in heterogeneous tissues with arbitrary geometries. The previous linear algorithm was modified to extract BFIs in different types of tissues simultaneously through utilizing DCS data at multiple source-detector separations. We compared the proposed linear algorithm with the semi-infinite homogenous solution in a computer model of adult head with heterogeneous tissue layers of scalp, skull, cerebrospinal fluid, and brain. To test the capability of the linear algorithm for extracting relative changes of cerebral blood flow (rCBF) in deep brain, we assigned ten levels of αD B in the brain layer with a step decrement of 10% while maintaining αD B values constant in other layers. Simulation results demonstrate the accuracy (errors model simplifies data analysis, thus allowing for online data processing and displaying. Future study will test this linear algorithm in heterogeneous tissues with different levels of blood flow variations and noises.

  13. Matrix effect on emission/current correlated analysis in laser-induced breakdown spectroscopy of liquid droplets

    Huang, J.-S.; Ke, C.-B.; Lin, K.-C.

    2004-01-01

    We have investigated influence of matrix salts on the liquid droplets by laser-induced breakdown spectroscopy (LIBS). An electrospray ionization technique coupled with LIBS is employed to generate the microdroplets of the Na sample solution with various matrix salts added. A sequence of single-shot time-resolved LIB emission signals is detected. The LIB signal intensity integrated within a gate linearly correlates with the plasma-induced current response obtained simultaneously on a single-shot basis. The slopes thus obtained increase with the sample concentration, but appear to be irrespective of different matrix salts, added up to a 2000 mg/l concentration. The matrix salts involved have the same K + cation but different anions. Given a laser radiation emitting at 355 nm with the energy fixed at 23±1 mJ, a limit of detection (LOD) of 1.0 mg/l may be achieved for the Na analysis. The current normalization might have probably taken into account the ablated amount of the sample and the plasma temperature. Accordingly, the LIB/current correlated analysis becomes efficient to suppress the signal fluctuation, improve the LOD determination, and concurrently correct the matrix effect

  14. Fluorescence Correlation Spectroscopy to find the critical balance between extracellular association and intracellular dissociation of mRNA-complexes.

    Zhang, Heyang; De Smedt, Stefaan C; Remaut, Katrien

    2018-05-10

    Fluorescence Correlation Spectroscopy (FCS) is a promising tool to study interactions on a single molecule level. The diffusion of fluorescent molecules in and out of the excitation volume of a confocal microscope leads to the fluorescence fluctuations that give information on the average number of fluorescent molecules present in the excitation volume and their diffusion coefficients. In this context, we complexed mRNA into lipoplexes and polyplexes and explored the association/dissociation degree of complexes by using gel electrophoresis and FCS. FCS enabled us to measure the association and dissociation degree of mRNA-based complexes both in buffer and protein-rich biological fluids such as human serum and ascitic fluid, which is a clear advantage over gel electrophoresis that was only applicable in protein-free buffer solutions. Furthermore, following the complex stability in buffer and biological fluids by FCS assisted to understand how complex characteristics, such as charge ratio and strength of mRNA binding, correlated to the transfection efficiency. We found that linear polyethyleneimine prevented efficient translation of mRNA, most likely due to a too strong mRNA binding, whereas the lipid based carrier Lipofectamine ® messengerMAX did succeed in efficient release and subsequent translation of mRNA in the cytoplasm of the cells. Overall, FCS is a reliable tool for the in depth characterization of mRNA complexes and can help us to find the critical balance keeping mRNA bound in complexes in the extracellular environment and efficient intracellular mRNA release leading to protein production. The delivery of messenger RNA (mRNA) to cells is promising to treat a variety of diseases. Therefore, the mRNA is typically packed in small lipid particles or polymer particles that help the mRNA to reach the cytoplasm of the cells. These particles should bind and carry the mRNA in the extracellular environment (e.g. blood, peritoneal fluid, ...), but should release

  15. Off-shell pairing correlations from meson-exchange theory of nuclear forces

    Sedrakian, Armen

    2003-01-01

    We develop a model of off-mass-shell pairing correlations in nuclear systems, which is based on the meson-exchange picture of nuclear interactions. The temporal retardations in the model are generated by the Fock-exchange diagrams. The kernel of the complex gap equation for baryons is related to the in-medium spectral function of mesons, which is evaluated nonperturbatively in the random phase approximation. The model is applied to the low-density neutron matter in neutron star crusts by separating the interaction into a long-range one-pion-exchange component and a short-range component parametrized in terms of Landau Fermi liquid parameters. The resulting Eliashberg-type coupled nonlinear integral equations are solved by an iterative procedure. We find that the self-energies extend to off-shell energies of the order of several tens of MeV. At low energies the damping of the neutron pair correlations due to the coupling to the pionic modes is small, but becomes increasingly important as the energy is increased. We discuss an improved quasiclassical approximation under which the numerical solutions are obtained

  16. Correlation Between Orthodontic Forces and Root Resorption – a Systematic Review of the Literature

    Vlasa Alexandru

    2016-09-01

    Full Text Available Orthodontically induced external apical root resorption (OIEARR is a major concern regarding periodontal status after nonsurgical orthodontic treatment. The aim of this study was to assess this sequel by a systematic review of published data. For assessment, we performed an electronic search of one database for comprehensive data, using keywords in different combinations: “root resorption”, “periodontics” and “nonsurgical orthodontic treatment”. We supplemented the results searching by hand in published journals and we cross-referenced with the accessed articles. Patients included in the results presented a good general health status, with no previous history of OIEARR and no other associated pathologies. Finally, twenty-three studies were selected and included in this review. A high prevalence (69–98% and moderate severity of OIEARR (<5 mm and <1/3 from original root length were reported. No difference in root resorption was found regarding the sex of the patients. A moderate positive correlation between treatment duration and root resorption was found. Also, a mild correlation regarding antero-posterior apical displacement and root resorption was found.

  17. Constitutive correlations for wire-wrapped subchannel analysis under forced and mixed convection conditions. Part 1

    Cheng, S.K.; Todreas, N.E.

    1984-08-01

    A simple subchannel analysis method based on the ENERGY series of codes, ENERGY-IV, has been established for predicting the temperature field in a single isolated wire-wrapped Liquid Metal Fast Breeder Reactor (LMFBR) subassembly under steady state forced and mixed convection conditions. The ENERGY-IV is a totally empirical code employed for fast running purposes and requires well calibrated lead length averaged input parameters to achieve satisfactory predictions. These input parameters were identified to be the inlet flow split parameters, the subchannel friction factors, the interchannel mixing parameters, the conduction shape factor, and the transverse velocity at the edge gap. Experiments were performed in a 37-pin wire-wrapped rod bundle with a geometry between that of a typical LMFBR fuel subassembly and blanket subassembly for filling the gap in the available data base for the input parameters. The isokinetic extraction method for measuring subchannel velocity, the pitot-static probe for measuring pressure drop, and the salt tracer injection method for estimating the interchannel mixing, were used in these experiments

  18. Correlation of Normal Gravity Mixed Convection Blowoff Limits with Microgravity Forced Flow Blowoff Limits

    Marcum, Jeremy W.; Olson, Sandra L.; Ferkul, Paul V.

    2016-01-01

    The axisymmetric rod geometry in upward axial stagnation flow provides a simple way to measure normal gravity blowoff limits to compare with microgravity Burning and Suppression of Solids - II (BASS-II) results recently obtained aboard the International Space Station. This testing utilized the same BASS-II concurrent rod geometry, but with the addition of normal gravity buoyant flow. Cast polymethylmethacrylate (PMMA) rods of diameters ranging from 0.635 cm to 3.81 cm were burned at oxygen concentrations ranging from 14 to 18% by volume. The forced flow velocity where blowoff occurred was determined for each rod size and oxygen concentration. These blowoff limits compare favorably with the BASS-II results when the buoyant stretch is included and the flow is corrected by considering the blockage factor of the fuel. From these results, the normal gravity blowoff boundary for this axisymmetric rod geometry is determined to be linear, with oxygen concentration directly proportional to flow speed. We describe a new normal gravity 'upward flame spread test' method which extrapolates the linear blowoff boundary to the zero stretch limit in order to resolve microgravity flammability limits-something current methods cannot do. This new test method can improve spacecraft fire safety for future exploration missions by providing a tractable way to obtain good estimates of material flammability in low gravity.

  19. Correlated Fluorescence-Atomic Force Microscopy Studies of the Clathrin Mediated Endocytosis in SKMEL Cells

    Smith, Steve; Hor, Amy; Luu, Anh; Kang, Lin; Scott, Brandon; Bailey, Elizabeth; Hoppe, Adam

    Clathrin-mediated endocytosis is one of the central pathways for cargo transport into cells, and plays a major role in the maintenance of cellular functions, such as intercellular signaling, nutrient intake, and turnover of plasma membrane in cells. The clathrin-mediated endocytosis process involves invagination and formation of clathrin-coated vesicles. However, the biophysical mechanisms of vesicle formation are still debated. We investigate clathrin vesicle formation mechanisms through the utilization of tapping-mode atomic force microscopy for high resolution topographical imaging in neutral buffer solution of unroofed cells exposing the inner membrane, combined with fluorescence imaging to definitively label intracellular constituents with specific fluorescent fusion proteins (actin filaments labeled with green phalloidin-antibody and clathrin coated vesicles with the fusion protein Tq2) in SKMEL (Human Melanoma) cells. Results from our work are compared against dynamical polarized total internal fluorescence (TIRF), super-resolution photo-activated localization microscopy (PALM) and transmission electron microscopy (TEM) to draw conclusions regarding the prominent model of vesicle formation in clathrin-mediated endocytosis. Funding provided by NSF MPS/DMR/BMAT award # 1206908.

  20. Correlation of the vapor pressure isotope effect with molecular force fields in the liquid state

    Pollin, J.S.; Ishida, T.

    1976-07-01

    The present work is concerned with the development and application of a new model for condensed phase interactions with which the vapor pressure isotope effect (vpie) may be related to molecular forces and structure. The model considers the condensed phase as being represented by a cluster of regularly arranged molecules consisting of a central molecule and a variable number of molecules in the first coordination shell. The methods of normal coordinate analysis are used to determine the modes of vibration of the condensed phase cluster from which, in turn, the isotopic reduced partition function can be calculated. Using the medium cluster model, the observed vpie for a series of methane isotopes has been successfully reproduced with better agreement with experiment than has been possible using the simple cell model. We conclude, however, that insofar as the medium cluster model provides a reasonable picture of the liquid state, the vpie is not sufficiently sensitive to molecular orientation to permit an experimental determination of intermolecular configuration in the condensed phase through measurement of isotopic pressure ratios. The virtual independence of vapor pressure isotope effects on molecular orientation at large cluster sizes is a demonstration of the general acceptability of the cell model assumptions for vpie calculations

  1. Neural correlates of dual-task effect on belief-bias syllogistic reasoning: a near-infrared spectroscopy study.

    Tsujii, Takeo; Watanabe, Shigeru

    2009-09-01

    Recent dual-process reasoning theories have explained the belief-bias effect, the tendency for human reasoning to be erroneously biased when logical conclusions are incongruent with beliefs about the world, by proposing a belief-based automatic heuristic system and logic-based demanding analytic system. Although these claims are supported by the behavioral finding that high-load secondary tasks enhance the belief-bias effect, the neural correlates of dual-task reasoning remain unknown. The present study therefore examined the relationship between dual-task effect and activity in the inferior frontal cortex (IFC) during belief-bias reasoning by near-infrared spectroscopy (NIRS). Forty-eight subjects participated in this study (MA=23.46 years). They were required to perform congruent and incongruent reasoning trials while responding to high- and low-load secondary tasks. Behavioral analysis showed that the high-load secondary task impaired only incongruent reasoning performance. NIRS analysis found that the high-load secondary task decreased right IFC activity during incongruent trials. Correlation analysis showed that subjects with enhanced right IFC activity could perform better in the incongruent reasoning trials, though subjects for whom right IFC activity was impaired by the secondary task could not maintain better reasoning performance. These findings suggest that the right IFC may be responsible for the dual-task effect in conflicting reasoning processes. When secondary tasks impair right IFC activity, subjects may rely on the automatic heuristic system, which results in belief-bias responses. We therefore offer the first demonstration of neural correlates of dual-task effect on IFC activity in belief-bias reasoning.

  2. Two-dimensional correlation infrared spectroscopy applied to analyzing and identifying the extracts of Baeckea frutescens medicinal materials.

    Adib, Adiana Mohamed; Jamaludin, Fadzureena; Kiong, Ling Sui; Hashim, Nuziah; Abdullah, Zunoliza

    2014-08-05

    Baeckea frutescens or locally known as Cucur atap is used as antibacterial, antidysentery, antipyretic and diuretic agent. In Malaysia and Indonesia, they are used as an ingredient of the traditional medicine given to mothers during confinement. A three-steps infra-red (IR) macro-fingerprinting method combining conventional IR spectra, and the secondary derivative spectra with two dimensional infrared correlation spectroscopy (2D-IR) have been proved to be effective methods to examine a complicated mixture such as herbal medicines. This study investigated the feasibility of employing multi-steps IR spectroscopy in order to study the main constituents of B. frutescens and its different extracts (extracted by chloroform, ethyl acetate, methanol and aqueous in turn). The findings indicated that FT-IR and 2D-IR can provide many holistic variation rules of chemical constituents. The structural information of the samples indicated that B. frutescens and its extracts contain a large amount of flavonoids, since some characteristic absorption peaks of flavonoids, such as ∼1600cm(-1), ∼1500cm(-1), ∼1450cm(-1), and ∼1270cm(-1) can be observed. The macroscopical fingerprint characters of FT-IR and 2D-IR spectra can not only provide the information of main chemical constituents in medicinal materials and their different extracts, but also compare the components differences among the similar samples. In conclusion, the multi-steps IR macro-fingerprint method is rapid, effective, visual and accurate for pharmaceutical research. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Hemodynamic measurements in rat brain and human muscle using diffuse near-infrared absorption and correlation spectroscopies

    Yu, Guoqiang; Durduran, Turgut; Furuya, D.; Lech, G.; Zhou, Chao; Chance, Britten; Greenberg, J. H.; Yodh, Arjun G.

    2003-07-01

    Measurement of concentration, oxygenation, and flow characteristics of blood cells can reveal information about tissue metabolism and functional heterogeneity. An improved multifunctional hybrid system has been built on the basis of our previous hybrid instrument that combines two near-infrared diffuse optical techniques to simultaneously monitor the changes of blood flow, total hemoglobin concentration (THC) and blood oxygen saturation (StO2). Diffuse correlation spectroscopy (DCS) monitors blood flow (BF) by measuring the optical phase shifts caused by moving blood cells, while diffuse photon density wave spectroscopy (DPDW) measures tissue absorption and scattering. Higher spatial resolution, higher data acquisition rate and higher dynamic range of the improved system allow us to monitor rapid hemodynamic changes in rat brain and human muscles. We have designed two probes with different source-detector pairs and different separations for the two types of experiments. A unique non-contact probe mounted on the back of a camera, which allows continuous measurements without altering the blood flow, was employed to in vivo monitor the metabolic responses in rat brain during KCl induced cortical spreading depression (CSD). A contact probe was used to measure changes of blood flow and oxygenation in human muscle during and after cuff occlusion or exercise, where the non-contact probe is not appropriate for monitoring the moving target. The experimental results indicate that our multifunctional hybrid system is capable of in vivo and non-invasive monitoring of the hemodynamic changes in different tissues (smaller tissues in rat brain, larger tissues in human muscle) under different conditions (static versus moving). The time series images of flow during CSD obtained by our technique revealed spatial and temporal hemodynamic changes in rat brain. Two to three fold longer recovery times of flow and oxygenation after cuff occlusion or exercise from calf flexors in a

  4. Single-Molecule Force Spectroscopy Trajectories of a Single Protein and Its Polyproteins Are Equivalent: A Direct Experimental Validation Based on A Small Protein NuG2.

    Lei, Hai; He, Chengzhi; Hu, Chunguang; Li, Jinliang; Hu, Xiaodong; Hu, Xiaotang; Li, Hongbin

    2017-05-22

    Single-molecule force spectroscopy (SMFS) has become a powerful tool in investigating the mechanical unfolding/folding of proteins at the single-molecule level. Polyproteins made of tandem identical repeats have been widely used in atomic force microscopy (AFM)-based SMFS studies, where polyproteins not only serve as fingerprints to identify single-molecule stretching events, but may also improve statistics of data collection. However, the inherent assumption of such experiments is that all the domains in the polyprotein are equivalent and one SMFS trajectory of stretching a polyprotein made of n domains is equivalent to n trajectories of stretching a single domain. Such an assumption has not been validated experimentally. Using a small protein NuG2 and its polyprotein (NuG2) 4 as model systems, here we use optical trapping (OT) to directly validate this assumption. Our results show that OT experiments on NuG2 and (NuG2) 4 lead to identical parameters describing the unfolding and folding kinetics of NuG2, demonstrating that indeed stretching a polyprotein of NuG2 is equivalent to stretching single NuG2 in force spectroscopy experiments and thus validating the use of polyproteins in SMFS experiments. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Gamow-Teller strength function for 90Zr: Effects of spin and isospin exchange forces, and ground-state correlations

    Mathews, G.J.; Bloom, S.D.; Hausman, R.F. Jr.

    1983-01-01

    Shell-model calculations of the Gamow-Teller strength function for 90 Zr have been performed utilizing a realistic finite-range two-body interaction in a model space consisting of the 2p and 1g shells. The effects of admixtures of two-particle two-hole excitations in 90 Nb, mostly due to the spin and isospin exchange components of the nucleon-nucleon force, are discussed. Ground state correlations in 90 Zr are also added via seniority-zero two-proton excitations from the 2p shell into the 1g/sub 9/2/ shell. With the correlations the Gamow-Teller strength function is in good agreement with the experimental results and accounts for essentially all of the observed dispersion of strength. The inclusion of these correlations does not, however, produce either a displacement of Gamow-Teller strength to higher excitation energies, or a significant change in the total strength. Thus, they cannot account for the observed Gamow-Teller quenching. The quenching factor derived by a comparison of our calculated results with experiment is 0.52

  6. Correlation between Onset Oxidation Temperature (OOT) and Fourier Transform Infrared Spectroscopy (FTIR) for monitoring the restabilization of Recycled Low-density Polyethylene (LDPE)

    Ruvolo-Filho, Adhemar; Pelozzi, Tadeu Luiz Alonso, E-mail: adhemar@power.ufscar.br [Universidade Federal de Sao Carlos (UFSCar), SP (Brazil). Departamento de Quimica

    2013-07-01

    In this work a new method was developed for monitoring the oxidative stability of restabilized and non-restabilized low-density polyethylene (LDPE) during multiple extrusion cycles. The method is based on correlations between Fourier Transform Infrared Spectroscopy (FTIR) and Onset Oxidation Temperature (OOT). Non-linear calibration curves correlating the concentration of primary or secondary antioxidants and the OOT values were obtained. (author)

  7. The diffusion dynamics of PEGylated liposomes in the intact vitreous of the ex vivo porcine eye: A fluorescence correlation spectroscopy and biodistribution study

    Eriksen, Anne Zebitz; Brewer, Jonathan; Andresen, Thomas Lars

    2017-01-01

    correlation spectroscopy (FCS) to determine liposome diffusion coefficients in the intact vitreous (DVit) of ex vivo porcine eyes using a modified Miyake-Apple technique to minimize the disruption of the vitreous fine structure. We chose to investigate whether the zeta potential of polyethylene glycol...

  8. A Nth-order linear algorithm for extracting diffuse correlation spectroscopy blood flow indices in heterogeneous tissues

    Shang, Yu; Yu, Guoqiang

    2014-01-01

    Conventional semi-infinite analytical solutions of correlation diffusion equation may lead to errors when calculating blood flow index (BFI) from diffuse correlation spectroscopy (DCS) measurements in tissues with irregular geometries. Very recently, we created an algorithm integrating a Nth-order linear model of autocorrelation function with the Monte Carlo simulation of photon migrations in homogenous tissues with arbitrary geometries for extraction of BFI (i.e., αD B ). The purpose of this study is to extend the capability of the Nth-order linear algorithm for extracting BFI in heterogeneous tissues with arbitrary geometries. The previous linear algorithm was modified to extract BFIs in different types of tissues simultaneously through utilizing DCS data at multiple source-detector separations. We compared the proposed linear algorithm with the semi-infinite homogenous solution in a computer model of adult head with heterogeneous tissue layers of scalp, skull, cerebrospinal fluid, and brain. To test the capability of the linear algorithm for extracting relative changes of cerebral blood flow (rCBF) in deep brain, we assigned ten levels of αD B in the brain layer with a step decrement of 10% while maintaining αD B values constant in other layers. Simulation results demonstrate the accuracy (errors < 3%) of high-order (N ≥ 5) linear algorithm in extracting BFIs in different tissue layers and rCBF in deep brain. By contrast, the semi-infinite homogenous solution resulted in substantial errors in rCBF (34.5% ≤ errors ≤ 60.2%) and BFIs in different layers. The Nth-order linear model simplifies data analysis, thus allowing for online data processing and displaying. Future study will test this linear algorithm in heterogeneous tissues with different levels of blood flow variations and noises.

  9. A Nth-order linear algorithm for extracting diffuse correlation spectroscopy blood flow indices in heterogeneous tissues

    Shang, Yu; Yu, Guoqiang, E-mail: guoqiang.yu@uky.edu [Department of Biomedical Engineering, University of Kentucky, Lexington, Kentucky 40506 (United States)

    2014-09-29

    Conventional semi-infinite analytical solutions of correlation diffusion equation may lead to errors when calculating blood flow index (BFI) from diffuse correlation spectroscopy (DCS) measurements in tissues with irregular geometries. Very recently, we created an algorithm integrating a Nth-order linear model of autocorrelation function with the Monte Carlo simulation of photon migrations in homogenous tissues with arbitrary geometries for extraction of BFI (i.e., αD{sub B}). The purpose of this study is to extend the capability of the Nth-order linear algorithm for extracting BFI in heterogeneous tissues with arbitrary geometries. The previous linear algorithm was modified to extract BFIs in different types of tissues simultaneously through utilizing DCS data at multiple source-detector separations. We compared the proposed linear algorithm with the semi-infinite homogenous solution in a computer model of adult head with heterogeneous tissue layers of scalp, skull, cerebrospinal fluid, and brain. To test the capability of the linear algorithm for extracting relative changes of cerebral blood flow (rCBF) in deep brain, we assigned ten levels of αD{sub B} in the brain layer with a step decrement of 10% while maintaining αD{sub B} values constant in other layers. Simulation results demonstrate the accuracy (errors < 3%) of high-order (N ≥ 5) linear algorithm in extracting BFIs in different tissue layers and rCBF in deep brain. By contrast, the semi-infinite homogenous solution resulted in substantial errors in rCBF (34.5% ≤ errors ≤ 60.2%) and BFIs in different layers. The Nth-order linear model simplifies data analysis, thus allowing for online data processing and displaying. Future study will test this linear algorithm in heterogeneous tissues with different levels of blood flow variations and noises.

  10. Spatio-temporal image correlation spectroscopy and super-resolution microscopy to quantify molecular dynamics in T cells.

    Ashdown, George W; Owen, Dylan M

    2018-02-02

    Many cellular processes are regulated by the spatio-temporal organisation of signalling complexes, cytoskeletal components and membranes. One such example is at the T cell immunological synapse where the retrograde flow of cortical filamentous (F)-actin from the synapse periphery drives signalling protein microclusters towards the synapse centre. The density of this mesh however, makes visualisation and analysis of individual actin fibres difficult due to the resolution limit of conventional microscopy. Recently, super-resolution methods such as structured illumination microscopy (SIM) have surpassed this resolution limit. Here, we apply SIM to better visualise the dense cortical actin meshwork in T cell synapses formed against activating, antibody-coated surfaces and image under total-internal reflection fluorescence (TIRF) illumination. To analyse the observed molecular flows, and the relationship between them, we apply spatio-temporal image correlation spectroscopy (STICS) and its cross-correlation variant (STICCS). We show that the dynamic cortical actin mesh can be visualised with unprecedented detail and that STICS/STICCS can output accurate, quantitative maps of molecular flow velocity and directionality from such data. We find that the actin flow can be disrupted using small molecule inhibitors of actin polymerisation. This combination of imaging and quantitative analysis may provide an important new tool for researchers to investigate the molecular dynamics at cellular length scales. Here we demonstrate the retrograde flow of F-actin which may be important for the clustering and dynamics of key signalling proteins within the plasma membrane, a phenomenon which is vital to correct T cell activation and therefore the mounting of an effective immune response. Copyright © 2018. Published by Elsevier Inc.

  11. Diffusion-weighted imaging and magnetic resonance spectroscopy of sporadic Creutzfeldt-Jakob disease: correlation with clinical course

    Kim, Jae Hyoung; Choi, Byung Se; Jung, Cheolkyu; Chang, YoungHee; Kim, SangYun

    2011-01-01

    Sporadic Creutzfeldt-Jakob disease (sCJD) is a fatal disease with variable clinical courses. The presence or absence of basal ganglia (BG) involvement has been reported to be associated with clinical course. We investigated the association of clinical course of sCJD with diffusion-weighted imaging (DWI) and MR spectroscopy (MRS) as well as BG involvement at early stage. DWI and single voxel proton MRS were performed in 14 patients with sCJD during the initial diagnostic workup. Apparent diffusion coefficient (ADC) and metabolites were measured in medial occipitoparietal cortices where large hyperintense DWI lesions were found in all patients. The presence or absence of BG involvement, ADC, N-acetylaspartate (NAA)/creatine (Cr) ratios, and choline (Cho)/Cr ratios were correlated with disease duration (i.e., the time from the symptom onset to death). The disease duration ranged from 2 to 31 months (median, 16). Hyperintense DWI lesions were observed bilaterally in both cortices and basal ganglia in eight patients and in cortices alone in six patients. Patients with BG involvement had shorter disease duration (median, 6.8 versus 20.5; p = 0.039) than those without and lower NAA/Cr ratios (median, 1.41 versus 2.03; p = 0.001). ADC and Cho/Cr ratios were not significantly different between the patients with BG involvement and those without. By multiple regression analysis, NAA/Cr ratios had the greatest correlation with the disease duration (p = 0.029). The disease duration of sCJD was variable. NAA/Cr ratios of the affected brain at the early stage of sCJD can be used as a useful parameter in predicting the clinical course. (orig.)

  12. Brain oxidative metabolism of the newborn dog: correlation between 31P NMR spectroscopy and pyridine nucleotide redox state.

    Mayevsky, A; Nioka, S; Subramanian, V H; Chance, B

    1988-04-01

    The effects of both anoxia and short- and long-term hypoxia on brain oxidative metabolism were studied in newborn dogs. Oxidative metabolism was evaluated by two independent measures: in vivo continuous monitoring of mitochondrial NADH redox state and energy stores as calculated from the phosphocreatine (PCr)/Pi levels measured by 31P nuclear magnetic resonance (NMR) spectroscopy. The hemodynamic response to low oxygen supply was further evaluated by measuring the changes in the reflected light intensity at 366 nm (the excitation wavelength for NADH). The animal underwent surgery and was prepared for monitoring of the two signals (NADH and PCr/Pi). It was then placed inside a Phosphoenergetics 260-80 NMR spectrometer magnet with a 31-cm bore. Each animal (1-21 days old) was exposed to short-term anoxia or hypoxia as well as to long-term hypoxia (1-2 h). The results can be summarized as follow: (a) In the normoxic brain, the ratio between PCr and Pi was greater than 1 (1.2-1.4), while under hypoxia or asphyxia a significant decrease that was correlated to the FiO2 levels was recorded. (b) A clear correlation was found between the decrease in PCr/Pi values and the increased NADH redox state developed under decreased O2 supply to the brain. (c) Exposing the animal to moderately long-term hypoxia led to a stabilized low-energy state of the brain with a good recovery after rebreathing normal air. (d) Under long-term and severe hypoxia, the microcirculatory autoregulatory mechanism was damaged and massive vasoconstriction was optically recorded simultaneously with a significant decrease in PCr/Pi values.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. A flexible fluorescence correlation spectroscopy based method for quantification of the DNA double labeling efficiency with precision control

    Hou, Sen; Tabaka, Marcin; Sun, Lili; Trochimczyk, Piotr; Kaminski, Tomasz S; Kalwarczyk, Tomasz; Zhang, Xuzhu; Holyst, Robert

    2014-01-01

    We developed a laser-based method to quantify the double labeling efficiency of double-stranded DNA (dsDNA) in a fluorescent dsDNA pool with fluorescence correlation spectroscopy (FCS). Though, for quantitative biochemistry, accurate measurement of this parameter is of critical importance, before our work it was almost impossible to quantify what percentage of DNA is doubly labeled with the same dye. The dsDNA is produced by annealing complementary single-stranded DNA (ssDNA) labeled with the same dye at 5′ end. Due to imperfect ssDNA labeling, the resulting dsDNA is a mixture of doubly labeled dsDNA, singly labeled dsDNA and unlabeled dsDNA. Our method allows the percentage of doubly labeled dsDNA in the total fluorescent dsDNA pool to be measured. In this method, we excite the imperfectly labeled dsDNA sample in a focal volume of <1 fL with a laser beam and correlate the fluctuations of the fluorescence signal to get the FCS autocorrelation curves; we express the amplitudes of the autocorrelation function as a function of the DNA labeling efficiency; we perform a comparative analysis of a dsDNA sample and a reference dsDNA sample, which is prepared by increasing the total dsDNA concentration c (c > 1) times by adding unlabeled ssDNA during the annealing process. The method is flexible in that it allows for the selection of the reference sample and the c value can be adjusted as needed for a specific study. We express the precision of the method as a function of the ssDNA labeling efficiency or the dsDNA double labeling efficiency. The measurement precision can be controlled by changing the c value. (letter)

  14. Dynamics-based selective 2D 1H/1H chemical shift correlation spectroscopy under ultrafast MAS conditions

    Zhang, Rongchun; Ramamoorthy, Ayyalusamy

    2015-05-01

    Dynamics plays important roles in determining the physical, chemical, and functional properties of a variety of chemical and biological materials. However, a material (such as a polymer) generally has mobile and rigid regions in order to have high strength and toughness at the same time. Therefore, it is difficult to measure the role of mobile phase without being affected by the rigid components. Herein, we propose a highly sensitive solid-state NMR approach that utilizes a dipolar-coupling based filter (composed of 12 equally spaced 90° RF pulses) to selectively measure the correlation of 1H chemical shifts from the mobile regions of a material. It is interesting to find that the rotor-synchronized dipolar filter strength decreases with increasing inter-pulse delay between the 90° pulses, whereas the dipolar filter strength increases with increasing inter-pulse delay under static conditions. In this study, we also demonstrate the unique advantages of proton-detection under ultrafast magic-angle-spinning conditions to enhance the spectral resolution and sensitivity for studies on small molecules as well as multi-phase polymers. Our results further demonstrate the use of finite-pulse radio-frequency driven recoupling pulse sequence to efficiently recouple weak proton-proton dipolar couplings in the dynamic regions of a molecule and to facilitate the fast acquisition of 1H/1H correlation spectrum compared to the traditional 2D NOESY (Nuclear Overhauser effect spectroscopy) experiment. We believe that the proposed approach is beneficial to study mobile components in multi-phase systems, such as block copolymers, polymer blends, nanocomposites, heterogeneous amyloid mixture of oligomers and fibers, and other materials.

  15. Dynamics-based selective 2D 1H/1H chemical shift correlation spectroscopy under ultrafast MAS conditions

    Zhang, Rongchun; Ramamoorthy, Ayyalusamy

    2015-01-01

    Dynamics plays important roles in determining the physical, chemical, and functional properties of a variety of chemical and biological materials. However, a material (such as a polymer) generally has mobile and rigid regions in order to have high strength and toughness at the same time. Therefore, it is difficult to measure the role of mobile phase without being affected by the rigid components. Herein, we propose a highly sensitive solid-state NMR approach that utilizes a dipolar-coupling based filter (composed of 12 equally spaced 90° RF pulses) to selectively measure the correlation of 1 H chemical shifts from the mobile regions of a material. It is interesting to find that the rotor-synchronized dipolar filter strength decreases with increasing inter-pulse delay between the 90° pulses, whereas the dipolar filter strength increases with increasing inter-pulse delay under static conditions. In this study, we also demonstrate the unique advantages of proton-detection under ultrafast magic-angle-spinning conditions to enhance the spectral resolution and sensitivity for studies on small molecules as well as multi-phase polymers. Our results further demonstrate the use of finite-pulse radio-frequency driven recoupling pulse sequence to efficiently recouple weak proton-proton dipolar couplings in the dynamic regions of a molecule and to facilitate the fast acquisition of 1 H/ 1 H correlation spectrum compared to the traditional 2D NOESY (Nuclear Overhauser effect spectroscopy) experiment. We believe that the proposed approach is beneficial to study mobile components in multi-phase systems, such as block copolymers, polymer blends, nanocomposites, heterogeneous amyloid mixture of oligomers and fibers, and other materials

  16. Diffusion-weighted imaging and magnetic resonance spectroscopy of sporadic Creutzfeldt-Jakob disease: correlation with clinical course

    Kim, Jae Hyoung; Choi, Byung Se; Jung, Cheolkyu [Seoul National University Bundang Hospital, Department of Radiology, Seoul National University College of Medicine, Seongnam-si (Korea, Republic of); Chang, YoungHee; Kim, SangYun [Seoul National University Bundang Hospital, Department of Neurology, Seoul National University College of Medicine, Seongnam-si (Korea, Republic of)

    2011-12-15

    Sporadic Creutzfeldt-Jakob disease (sCJD) is a fatal disease with variable clinical courses. The presence or absence of basal ganglia (BG) involvement has been reported to be associated with clinical course. We investigated the association of clinical course of sCJD with diffusion-weighted imaging (DWI) and MR spectroscopy (MRS) as well as BG involvement at early stage. DWI and single voxel proton MRS were performed in 14 patients with sCJD during the initial diagnostic workup. Apparent diffusion coefficient (ADC) and metabolites were measured in medial occipitoparietal cortices where large hyperintense DWI lesions were found in all patients. The presence or absence of BG involvement, ADC, N-acetylaspartate (NAA)/creatine (Cr) ratios, and choline (Cho)/Cr ratios were correlated with disease duration (i.e., the time from the symptom onset to death). The disease duration ranged from 2 to 31 months (median, 16). Hyperintense DWI lesions were observed bilaterally in both cortices and basal ganglia in eight patients and in cortices alone in six patients. Patients with BG involvement had shorter disease duration (median, 6.8 versus 20.5; p = 0.039) than those without and lower NAA/Cr ratios (median, 1.41 versus 2.03; p = 0.001). ADC and Cho/Cr ratios were not significantly different between the patients with BG involvement and those without. By multiple regression analysis, NAA/Cr ratios had the greatest correlation with the disease duration (p = 0.029). The disease duration of sCJD was variable. NAA/Cr ratios of the affected brain at the early stage of sCJD can be used as a useful parameter in predicting the clinical course. (orig.)

  17. Neural correlates of own- and other-race face recognition in children: a functional near-infrared spectroscopy study.

    Ding, Xiao Pan; Fu, Genyue; Lee, Kang

    2014-01-15

    The present study used the functional Near-infrared Spectroscopy (fNIRS) methodology to investigate the neural correlates of elementary school children's own- and other-race face processing. An old-new paradigm was used to assess children's recognition ability of own- and other-race faces. FNIRS data revealed that other-race faces elicited significantly greater [oxy-Hb] changes than own-race faces in the right middle frontal gyrus and inferior frontal gyrus regions (BA9) and the left cuneus (BA18). With increased age, the [oxy-Hb] activity differences between own- and other-race faces, or the neural other-race effect (NORE), underwent significant changes in these two cortical areas: at younger ages, the neural response to the other-race faces was modestly greater than that to the own-race faces, but with increased age, the neural response to the own-race faces became increasingly greater than that to the other-race faces. Moreover, these areas had strong regional functional connectivity with a swath of the cortical regions in terms of the neural other-race effect that also changed with increased age. We also found significant and positive correlations between the behavioral other-race effect (reaction time) and the neural other-race effect in the right middle frontal gyrus and inferior frontal gyrus regions (BA9). These results taken together suggest that children, like adults, devote different amounts of neural resources to processing own- and other-race faces, but the size and direction of the neural other-race effect and associated functional regional connectivity change with increased age. © 2013.

  18. Extraction of diffuse correlation spectroscopy flow index by integration of Nth-order linear model with Monte Carlo simulation

    Shang, Yu; Li, Ting; Chen, Lei; Lin, Yu; Toborek, Michal; Yu, Guoqiang

    2014-05-01

    Conventional semi-infinite solution for extracting blood flow index (BFI) from diffuse correlation spectroscopy (DCS) measurements may cause errors in estimation of BFI (αDB) in tissues with small volume and large curvature. We proposed an algorithm integrating Nth-order linear model of autocorrelation function with the Monte Carlo simulation of photon migrations in tissue for the extraction of αDB. The volume and geometry of the measured tissue were incorporated in the Monte Carlo simulation, which overcome the semi-infinite restrictions. The algorithm was tested using computer simulations on four tissue models with varied volumes/geometries and applied on an in vivo stroke model of mouse. Computer simulations shows that the high-order (N ≥ 5) linear algorithm was more accurate in extracting αDB (errors values of errors in extracting αDB were similar to those reconstructed from the noise-free DCS data. In addition, the errors in extracting the relative changes of αDB using both linear algorithm and semi-infinite solution were fairly small (errors < ±2.0%) and did not rely on the tissue volume/geometry. The experimental results from the in vivo stroke mice agreed with those in simulations, demonstrating the robustness of the linear algorithm. DCS with the high-order linear algorithm shows the potential for the inter-subject comparison and longitudinal monitoring of absolute BFI in a variety of tissues/organs with different volumes/geometries.

  19. Cross-correlation of motor activity signals from dc-magnetoencephalography, near-infrared spectroscopy, and electromyography.

    Sander, Tilmann H; Leistner, Stefanie; Wabnitz, Heidrun; Mackert, Bruno-Marcel; Macdonald, Rainer; Trahms, Lutz

    2010-01-01

    Neuronal and vascular responses due to finger movements were synchronously measured using dc-magnetoencephalography (dcMEG) and time-resolved near-infrared spectroscopy (trNIRS). The finger movements were monitored with electromyography (EMG). Cortical responses related to the finger movement sequence were extracted by independent component analysis from both the dcMEG and the trNIRS data. The temporal relations between EMG rate, dcMEG, and trNIRS responses were assessed pairwise using the cross-correlation function (CCF), which does not require epoch averaging. A positive lag on a scale of seconds was found for the maximum of the CCF between dcMEG and trNIRS. A zero lag is observed for the CCF between dcMEG and EMG. Additionally this CCF exhibits oscillations at the frequency of individual finger movements. These findings show that the dcMEG with a bandwidth up to 8 Hz records both slow and faster neuronal responses, whereas the vascular response is confirmed to change on a scale of seconds.

  20. Diffuse correlation spectroscopy (DCS) study of blood flow changes during low level laser therapy (LLLT): a preliminary report

    Soni, Sagar; Wang, Xinlong; Liu, Hanli; Tian, Fenghua

    2017-02-01

    Photobiomodulation with low-power, high-fluence light in the near-infrared range (600-1100nm), also known as low level laser therapy (LLLT), has been used for promoting healing of wounds, reducing pain, and so on. Understanding its physiological effect is essential for treatment optimization and evaluation. In this study, we used diffuse correlation spectroscopy (DCS) to investigate the changes of regional blood flow in skeletal muscle induced by a single session of LLLT. DCS is an emerging optical modality to probe microvascular blood flow in human tissues in vivo. We have developed a software-based autocorrelator system with the benefits such as flexibility in raw photon count data processing, portability and low cost. LLLT was administered at the human forearm with a 1064-nm, continuous-wave laser. The emitting power was 3.4 W in an area of 13.6 cm2, corresponding to 0.25W/cm2 irradiance. The emitting duration was 10 minutes. Eight healthy adults of any ethnic background, in an age range of 18-40 years old were included. The results indicate that LLLT causes reliable changes in regional blood flow. However, it remains unclear whether these changes are physiological or attributed to the heating effect of the stimulation laser.

  1. Effects of surface functionalization on the adsorption of human serum albumin onto nanoparticles – a fluorescence correlation spectroscopy study

    Pauline Maffre

    2014-11-01

    Full Text Available By using fluorescence correlation spectroscopy (FCS, we have studied the adsorption of human serum albumin (HSA onto Fe–Pt nanoparticles (NPs, 6 nm radius, CdSe/ZnS quantum dots (QDs, 5 nm radius and Au and Ag nanoclusters (1–4 nm radius, which are enshrouded by various water-solubilizing surface layers exposing different chemical functional groups (carboxyl, amino and both, thereby endowing the NPs with different surface charges. We have also measured the effects of modified surface functionalizations on the protein via succinylation and amination. A step-wise increase in hydrodynamic radius with protein concentration was always observed, revealing formation of protein monolayers coating the NPs, independent of their surface charge. The differences in the thickness of the protein corona were rationalized in terms of the different orientations in which HSA adsorbs onto the NPs. The midpoints of the binding transition, which quantifies the affinity of HSA toward the NP, were observed to differ by almost four orders of magnitude. These variations can be understood in terms of specific Coulombic interactions between the proteins and the NP surfaces.

  2. Correlations between Diffusion Tensor Imaging (DTI and Magnetic Resonance Spectroscopy (1H MRS in schizophrenic patients and normal controls

    Ng Johnny

    2007-06-01

    Full Text Available Abstract Background Evidence suggests that white matter integrity may play an underlying pathophysiological role in schizophrenia. N-acetylaspartate (NAA, as measured by Magnetic Resonance Spectroscopy (MRS, is a neuronal marker and is decreased in white matter lesions and regions of axonal loss. It has also been found to be reduced in the prefrontal and temporal regions in patients with schizophrenia. Diffusion Tensor Imaging (DTI allows one to measure the orientations of axonal tracts as well as the coherence of axonal bundles. DTI is thus sensitive to demyelination and other structural abnormalities. DTI has also shown abnormalities in these regions. Methods MRS and DTI were obtained on 42 healthy subjects and 40 subjects with schizophrenia. The data was analyzed using regions of interests in the Dorso-Lateral Prefrontal white matter, Medial Temporal white matter and Occipital white matter using both imaging modalities. Results NAA was significantly reduced in the patient population in the Medial Temporal regions. DTI anisotropy indices were also reduced in the same Medial Temporal regions. NAA and DTI-anisotropy indices were also correlated in the left medial temporal region. Conclusion Our results implicate defects in the medial temporal white matter in patients with schizophrenia. Moreover, MRS and DTI are complementary modalities for the study of white matter disruptions in patients with schizophrenia.

  3. Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data.

    Ashdown, George; Pandžić, Elvis; Cope, Andrew; Wiseman, Paul; Owen, Dylan

    2015-12-17

    Filamentous-actin plays a crucial role in a majority of cell processes including motility and, in immune cells, the formation of a key cell-cell interaction known as the immunological synapse. F-actin is also speculated to play a role in regulating molecular distributions at the membrane of cells including sub-membranous vesicle dynamics and protein clustering. While standard light microscope techniques allow generalized and diffraction-limited observations to be made, many cellular and molecular events including clustering and molecular flow occur in populations at length-scales far below the resolving power of standard light microscopy. By combining total internal reflection fluorescence with the super resolution imaging method structured illumination microscopy, the two-dimensional molecular flow of F-actin at the immune synapse of T cells was recorded. Spatio-temporal image correlation spectroscopy (STICS) was then applied, which generates quantifiable results in the form of velocity histograms and vector maps representing flow directionality and magnitude. This protocol describes the combination of super-resolution imaging and STICS techniques to generate flow vectors at sub-diffraction levels of detail. This technique was used to confirm an actin flow that is symmetrically retrograde and centripetal throughout the periphery of T cells upon synapse formation.

  4. Probing Temperature- and pH-Dependent Binding between Quantum Dots and Bovine Serum Albumin by Fluorescence Correlation Spectroscopy

    Zonghua Wang

    2017-04-01

    Full Text Available Luminescent quantum dots (QDs with unique optical properties have potential applications in bio-imaging. The interaction between QDs and bio-molecules is important to the biological effect of QDs in vivo. In this paper, we have employed fluorescence correlation spectroscopy (FCS to probe the temperature- and pH-dependent interactions between CdSe QDs with carboxyl (QDs-COOH and bovine serum albumin (BSA in buffer solutions. The results have shown that microscopic dissociation constant K′D is in the range of (1.5 ± 0.2 × 10−5 to (8.6 ± 0.1 × 10−7 M, the Hill coefficient n is from 0.4 to 2.3, and the protein corona thickness is from 3.0 to 9.4 nm. Variable-temperature measurements have shown both negative values of ∆H and ∆S for BSA adsorption on QDs-COOH, while pH has a profound effect on the adsorption. Additional, FCS measurement QDs-COOH and proteins in whole mice serum and plasma samples has also been conducted. Finally, simulation results have shown four favored QD binding sites in BSA.

  5. The Effect of a Fluorophore Photo-Physics on the Lipid Vesicle Diffusion Coefficient Studied by Fluorescence Correlation Spectroscopy.

    Drabik, Dominik; Przybyło, Magda; Sikorski, Aleksander; Langner, Marek

    2016-03-01

    Fluorescence Correlation Spectroscopy (FCS) is a technique, which allows determination of the diffusion coefficient and concentration of fluorescent objects suspended in the solution. The measured parameter is the fluctuation of the fluorescence signal emitted by diffusing molecules. When 100 nm DOPC vesicles labeled with various fluorescent dyes (Fluorescein-PE, NBD-PE, Atto488 DOPE or βBodipy FL) were measured, different values of diffusion coefficients have been obtained. These diffusion coefficients were different from the expected values measured using the dynamic light scattering method (DLS). The FCS was initially developed for solutions containing small fluorescent molecules therefore the observed inconsistency may result from the nature of vesicle suspension itself. The duration of the fluorescence signal may depend on the following factors: the exposure time of the labeled object to the excitation beam, the photo-physical properties (e.g., stability) of a fluorophore, the theoretical model used for the calculations of the diffusion coefficient and optical properties of the vesicle suspension. The diffusion coefficients determined for differently labeled liposomes show that its dependence on vesicle size and quantity of fluorescent probed used for labeling was significant demonstrating that the fluorescence properties of the fluorophore itself (bleaching and/or blinking) were critical factors for a correct outcome of FCS experiment. The new, based on combined FCS and DLS measurements, method for the determination of the focal volume prove itself to be useful for the evaluation of a fluorescence dye with respect to its applicability for FCS experiment.

  6. Evaluation of invasiveness of astrocytoma using 1H-magnetic resonance spectroscopy: correlation with expression of matrix metalloproteinase-2

    Zhang, Kai; Li, Chuanfu; Ma, Xiangxing; Meng, Xiangshui; Feng, Dechao; Liu, Ying; Li, Li

    2007-01-01

    Even low-grade astrocytomas infiltrate the entire brain, a feature that precludes their successful therapy. So to assess the invasive potential of astrocytoma is very important. The aim of this study was determine whether there is a significant correlation between the results of 1 H-magnetic resonance spectroscopy ( 1 H-MRS) and tumor invasive potential of astrocytoma, which is reflected by expression of matrix metalloproteinase-2 (MMP-2). The 1 H-MRS spectra of 41 histologically verified astrocytomas were obtained on a 3-T MR scanner. According to the World Health Organization classification criteria for central nervous system tumors, there were 16 low-grade astrocytomas (2 pilocytic astrocytomas, 14 grade II astrocytomas) and 25 high-grade astrocytomas (5 anaplastic astrocytomas, 20 glioblastomas).The choline/N-acetylaspartate (Cho/NAA) and choline/creatine (Cho/Cr) ratios were calculated. Of the 41 astrocytomas, 19 (8 low-grade and 11 high-grade) were analyzed immunohistochemically. Expression of MMP-2 was determined using streptavidin-peroxidase complex (SP) staining which was quantified by calculating its calibrated opacity density (COD) using an image analysis system. The correlations between metabolite ratios and the quantitative data from the immunohistochemical tests in the 19 astrocytomas were determined. The Cho/NAA and Cho/Cr ratios of high-grade astrocytoma were both significantly greater than those of low-grade astrocytoma (t = -6.222, P = 0.000; t = -6.533, P = 0.000, respectively). MMP-2 COD values of high-grade astrocytomas were also significantly greater than those of low-grade astrocytomas (t = -5.892, P 0.000). There were strong positive correlations between Cho/NAA ratio and MMP-2 COD (r = 0.669, P = 0.002), and between Cho/Cr ratio and MMP-2 COD (r = 0.689, P = 0.001). 1 H-MRS is helpful in evaluating the invasiveness of astrocytomas and predicting prognosis preoperatively by determining the Cho/NAA and Cho/Cr ratios. (orig.)

  7. Dynamics of shearing force and its correlations with chemical compositions and in vitro dry matter digestibility of stylo (Stylosanthes guianensis) stem.

    Zi, Xuejuan; Li, Mao; Zhou, Hanlin; Tang, Jun; Cai, Yimin

    2017-12-01

    The study explored the dynamics of shearing force and its correlation with chemical compositions and in vitro dry matter digestibility (IVDMD) of stylo. The shearing force, diameter, linear density, chemical composition, and IVDMD of different height stylo stem were investigated. Linear regression analysis was done to determine the relationships between the shearing force and cut height, diameter, chemical composition, or IVDMD. The results showed that shearing force of stylo stem increased with plant height increasing and the crude protein (CP) content and IVDMD decreased but fiber content increased over time, resulting in decreased forage value. In addition, tall stem had greater shearing force than short stem. Moreover, shearing force is positively correlated with stem diameter, linear density and fiber fraction, but negatively correlated with CP content and IVDMD. Overall, shearing force is an indicator more direct, easier and faster to measure than chemical composition and digestibility for evaluation of forage nutritive value related to animal performance. Therefore, it can be used to evaluate the nutritive value of stylo.

  8. Study of dynamic behavior of EDTA molecule in solution using perturbed gamma-gamma angular correlation spectroscopy

    Amaral, Antonio A.; Silva, Andreia dos S.; Carbonari, Arthur W.; Lapolli, Andre L.

    2009-01-01

    In this work, PAC spectroscopy has been used to obtain the hyperfine parameters in EDTA molecules in solutions with pH 4.3 and pH 10.5 both measured at 77 K and 295 K using 181 Hf( 181 Ta) as probe nuclei. Both dynamic and static interactions were measured in aqueous solution, crystallized and re-hydrated samples in order to examine the motion and structure of EDTA-molecules. The hyperfine parameters, quadrupole interaction frequency (ν Q ), asymmetry (η), and the dynamic interaction frequency (λ) were obtained. The outcomes show that the rotational correlation time (τ CR ) is larger than the half-life of the intermediate state of probe nuclei. For samples with pH 4.3 and pH 10.5, it was observed an increase in ν Q when the temperature decreases, as expected, and also a variation of η, which is an evidence of a change in the EDTA molecule structure. 181 Hf is bound only to a single molecule site when the pH was 4.3, differently from the results for pH 10.5 sample, which showed two fractions with different ν Q indicating the possibility of 181 Hf being bonded to two different sites of the molecule. Measurements of the dehydrated sample presented different results leading us to conclude that the preparation procedure can causes alterations in the chemical bounds. Concluding, these results showed a systematic behavior of the 181 Hf-EDTA, with the variation of pH from 4 to approximately 11, and they are important to the knowledge of the dynamic behavior of this molecule. (author)

  9. Unfolding of hemoglobin variants--insights from urea gradient gel electrophoresis photon correlation spectroscopy and zeta potential measurements

    Bhattacharya, Jaydeep; GhoshMoulick, Ranjita; Choudhuri, Utpal; Chakrabarty, Prantar; Bhattacharya, Pranab K.; Lahiri, Prabir; Chakraborti, Bikas; Dasgupta, Anjan Kr.

    2004-01-01

    The unfolding pattern of crystal human hemoglobin and variants of hemoglobin obtained from hemolysate were studied using transverse urea gradient gel electrophoresis (TUGGE). A smooth sigmoid like increase of electrophoretic mobility was observed with increasing urea concentrations. A decrease in electrophoretic mobility resulted, if the protein was unfolded with guanidium hydrochloride (GdnHCl). The anomaly was resolved after the Stoke's radii (obtained using the photon correlation spectroscopy) and zeta potential (measured using laser Doppler velocimetry) measurements were made at different denaturant concentrations. Addition of denaturant led to formation of extended structure, irrespective of the nature of the denaturant, as indicated by increase in Stoke's radii in both cases (urea and GdnHCl). The unexpected increase in electrophoretic mobility in case of urea could be explained in terms of a critical redistribution of negative charge at intermediate stages of the unfolding process. In case of GdnHCl, the higher ionic strength masked the charge effect. The mobility, being solely dependent on size, decreased at higher denaturant concentration. Incidentally, folding loci of other hemoglobin variants (e.g. HbE) or that of post-translationally modified hemoglobin (e.g. HbA1c) could be determined by studying the charge distribution and hydrodynamic radius at varying denaturing stress and in each case the gel migration profile could be approximately scaled by the ratio of charge and hydrodynamic diameter of the protein. While unfolding induced charge effect was most pronounced in HbA0 (and crystal ferrous hemoglobin), the unfolding induced aggregation (manifested by the increase in Stoke's radii) was predominantly observed in the variant forms HbE and HbA1c. Representing the proteins by a plot, in which charge and hydrodynamic diameter are on independent axes, may be a useful way of characterizing protein variants having similar migration profiles on native gels

  10. Unfolding of hemoglobin variants--insights from urea gradient gel electrophoresis photon correlation spectroscopy and zeta potential measurements

    Bhattacharya, Jaydeep; GhoshMoulick, Ranjita; Choudhuri, Utpal; Chakrabarty, Prantar; Bhattacharya, Pranab K.; Lahiri, Prabir; Chakraborti, Bikas; Dasgupta, Anjan Kr

    2004-09-27

    The unfolding pattern of crystal human hemoglobin and variants of hemoglobin obtained from hemolysate were studied using transverse urea gradient gel electrophoresis (TUGGE). A smooth sigmoid like increase of electrophoretic mobility was observed with increasing urea concentrations. A decrease in electrophoretic mobility resulted, if the protein was unfolded with guanidium hydrochloride (GdnHCl). The anomaly was resolved after the Stoke's radii (obtained using the photon correlation spectroscopy) and zeta potential (measured using laser Doppler velocimetry) measurements were made at different denaturant concentrations. Addition of denaturant led to formation of extended structure, irrespective of the nature of the denaturant, as indicated by increase in Stoke's radii in both cases (urea and GdnHCl). The unexpected increase in electrophoretic mobility in case of urea could be explained in terms of a critical redistribution of negative charge at intermediate stages of the unfolding process. In case of GdnHCl, the higher ionic strength masked the charge effect. The mobility, being solely dependent on size, decreased at higher denaturant concentration. Incidentally, folding loci of other hemoglobin variants (e.g. HbE) or that of post-translationally modified hemoglobin (e.g. HbA1c) could be determined by studying the charge distribution and hydrodynamic radius at varying denaturing stress and in each case the gel migration profile could be approximately scaled by the ratio of charge and hydrodynamic diameter of the protein. While unfolding induced charge effect was most pronounced in HbA0 (and crystal ferrous hemoglobin), the unfolding induced aggregation (manifested by the increase in Stoke's radii) was predominantly observed in the variant forms HbE and HbA1c. Representing the proteins by a plot, in which charge and hydrodynamic diameter are on independent axes, may be a useful way of characterizing protein variants having similar migration profiles on

  11. Mutational definition of binding requirements of an hnRNP-like protein in Arabidopsis using fluorescence correlation spectroscopy

    Leder, Verena [Molecular Cell Physiology, Faculty of Biology, Bielefeld University (Germany); Biomolecular Photonics, Faculty of Physics, Bielefeld University (Germany); Lummer, Martina [Molecular Cell Physiology, Faculty of Biology, Bielefeld University (Germany); Tegeler, Kathrin [Molecular Cell Physiology, Faculty of Biology, Bielefeld University (Germany); Biomolecular Photonics, Faculty of Physics, Bielefeld University (Germany); Humpert, Fabian [Biomolecular Photonics, Faculty of Physics, Bielefeld University (Germany); Lewinski, Martin [Molecular Cell Physiology, Faculty of Biology, Bielefeld University (Germany); Schüttpelz, Mark [Biomolecular Photonics, Faculty of Physics, Bielefeld University (Germany); Staiger, Dorothee, E-mail: dorothee.staiger@uni-bielefeld.de [Molecular Cell Physiology, Faculty of Biology, Bielefeld University (Germany)

    2014-10-10

    Highlights: • We use FCS to investigate binding site requirements for the hnRNP-like protein AtGRP7. • We identify three nucleotides critical for AtGRP7 binding to its own intron. • Mutation of the conserved R{sup 49} abolishes binding altogether. • The paralogue AtGRP8 binds to an overlapping motif with different sequence requirement. • The glycine-rich stretch of a plant hnRNP-like protein contributes to binding. - Abstract: Arabidopsis thaliana glycine-rich RNA binding protein 7 (AtGRP7) is part of a negative feedback loop through which it regulates alternative splicing and steady-state abundance of its pre-mRNA. Here we use fluorescence correlation spectroscopy to investigate the requirements for AtGRP7 binding to its intron using fluorescently-labelled synthetic oligonucleotides. By systematically introducing point mutations we identify three nucleotides that lead to an increased K{sub d} value when mutated and thus are critical for AtGRP7 binding. Simultaneous mutation of all three residues abrogates binding. The paralogue AtGRP8 binds to an overlapping motif but with a different sequence preference, in line with overlapping but not identical functions of this protein pair. Truncation of the glycine-rich domain reduces the binding affinity of AtGRP7, showing for the first time that the glycine-rich stretch of a plant hnRNP-like protein contributes to binding. Mutation of the conserved R{sup 49} that is crucial for AtGRP7 function in pathogen defence and splicing abolishes binding.

  12. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  13. PCR-free detection of genetically modified organisms using magnetic capture technology and fluorescence cross-correlation spectroscopy.

    Xiaoming Zhou

    2009-11-01

    Full Text Available The safety of genetically modified organisms (GMOs has attracted much attention recently. Polymerase chain reaction (PCR amplification is a common method used in the identification of GMOs. However, a major disadvantage of PCR is the potential amplification of non-target DNA, causing false-positive identification. Thus, there remains a need for a simple, reliable and ultrasensitive method to identify and quantify GMO in crops. This report is to introduce a magnetic bead-based PCR-free method for rapid detection of GMOs using dual-color fluorescence cross-correlation spectroscopy (FCCS. The cauliflower mosaic virus 35S (CaMV35S promoter commonly used in transgenic products was targeted. CaMV35S target was captured by a biotin-labeled nucleic acid probe and then purified using streptavidin-coated magnetic beads through biotin-streptavidin linkage. The purified target DNA fragment was hybridized with two nucleic acid probes labeled respectively by Rhodamine Green and Cy5 dyes. Finally, FCCS was used to detect and quantify the target DNA fragment through simultaneously detecting the fluorescence emissions from the two dyes. In our study, GMOs in genetically engineered soybeans and tomatoes were detected, using the magnetic bead-based PCR-free FCCS method. A detection limit of 50 pM GMOs target was achieved and PCR-free detection of GMOs from 5 microg genomic DNA with magnetic capture technology was accomplished. Also, the accuracy of GMO determination by the FCCS method is verified by spectrophotometry at 260 nm using PCR amplified target DNA fragment from GM tomato. The new method is rapid and effective as demonstrated in our experiments and can be easily extended to high-throughput and automatic screening format. We believe that the new magnetic bead-assisted FCCS detection technique will be a useful tool for PCR-free GMOs identification and other specific nucleic acids.

  14. Analysis of the substrate recognition state of TDP-43 to single-stranded DNA using fluorescence correlation spectroscopy

    Akira Kitamura

    2018-07-01

    Full Text Available Normal function and abnormal aggregation of transactivation response (TAR DNA/RNA-binding protein 43 kDa (TDP-43 are directly associated with the lethal genetic diseases: cystic fibrosis, amyotrophic lateral sclerosis (ALS, and frontotemporal lobar degeneration (FTLD. The binding of TDP-43 to single-stranded DNA (ssDNA or RNA is involved in transcriptional repression, regulation of RNA splicing, and RNA stabilization. Equilibrium dissociation constants (Kd of TDP-43 and ssDNA or RNA have been determined using various methods; however, methods that can measure Kd with high sensitivity in a short time using a small amount of TDP-43 in solution would be advantageous. Here, in order to determine the Kd of TDP-43 and fluorescence-labeled ssDNA as well as the binding stoichiometry, we use fluorescence correlation spectroscopy (FCS, which detects the slowed diffusion of molecular interactions in solution with single-molecule sensitivity, in addition to electrophoretic mobility shift assay (EMSA. Using tandem affinity chromatography of TDP-43 dually tagged with glutathione-S-transferase and poly-histidine tags, highly purified protein was obtained. FCS successfully detected specific interaction between purified TDP-43 and TG ssDNA repeats, with a Kd in the nanomolar range. The Kd of the TDP-43 mutant was not different from the wild type, although mutant oligomers, which did not bind ssDNA, were observed. Analysis of the fluorescence brightness per dimerized TDP-43/ssDNA complex was used to evaluate their binding stoichiometry. The results suggest that an assay combining FCS and EMSA can precisely analyze ssDNA recognition mechanisms, and that FCS may be applied for the rapid and quantitative determination of the interaction strength between TDP-43 and ssDNA or RNA. These methods will aid in the elucidation of the substrate recognition mechanism of ALS- and FTLD-associated variants of TDP-43.

  15. Mutational definition of binding requirements of an hnRNP-like protein in Arabidopsis using fluorescence correlation spectroscopy

    Leder, Verena; Lummer, Martina; Tegeler, Kathrin; Humpert, Fabian; Lewinski, Martin; Schüttpelz, Mark; Staiger, Dorothee

    2014-01-01

    Highlights: • We use FCS to investigate binding site requirements for the hnRNP-like protein AtGRP7. • We identify three nucleotides critical for AtGRP7 binding to its own intron. • Mutation of the conserved R 49 abolishes binding altogether. • The paralogue AtGRP8 binds to an overlapping motif with different sequence requirement. • The glycine-rich stretch of a plant hnRNP-like protein contributes to binding. - Abstract: Arabidopsis thaliana glycine-rich RNA binding protein 7 (AtGRP7) is part of a negative feedback loop through which it regulates alternative splicing and steady-state abundance of its pre-mRNA. Here we use fluorescence correlation spectroscopy to investigate the requirements for AtGRP7 binding to its intron using fluorescently-labelled synthetic oligonucleotides. By systematically introducing point mutations we identify three nucleotides that lead to an increased K d value when mutated and thus are critical for AtGRP7 binding. Simultaneous mutation of all three residues abrogates binding. The paralogue AtGRP8 binds to an overlapping motif but with a different sequence preference, in line with overlapping but not identical functions of this protein pair. Truncation of the glycine-rich domain reduces the binding affinity of AtGRP7, showing for the first time that the glycine-rich stretch of a plant hnRNP-like protein contributes to binding. Mutation of the conserved R 49 that is crucial for AtGRP7 function in pathogen defence and splicing abolishes binding

  16. Complementary studies of lipid membrane dynamics using iSCAT and super-resolved fluorescence correlation spectroscopy

    Reina, Francesco; Galiani, Silvia; Shrestha, Dilip; Sezgin, Erdinc; de Wit, Gabrielle; Cole, Daniel; Lagerholm, B. Christoffer; Kukura, Philipp; Eggeling, Christian

    2018-06-01

    Observation techniques with high spatial and temporal resolution, such as single-particle tracking based on interferometric scattering (iSCAT) microscopy, and fluorescence correlation spectroscopy applied on a super-resolution STED microscope (STED-FCS), have revealed new insights of the molecular organization of membranes. While delivering complementary information, there are still distinct differences between these techniques, most prominently the use of fluorescent dye tagged probes for STED-FCS and a need for larger scattering gold nanoparticle tags for iSCAT. In this work, we have used lipid analogues tagged with a hybrid fluorescent tag–gold nanoparticle construct, to directly compare the results from STED-FCS and iSCAT measurements of phospholipid diffusion on a homogeneous supported lipid bilayer (SLB). These comparative measurements showed that while the mode of diffusion remained free, at least at the spatial (>40 nm) and temporal (50  ⩽  t  ⩽  100 ms) scales probed, the diffussion coefficient was reduced by 20- to 60-fold when tagging with 20 and 40 nm large gold particles as compared to when using dye tagged lipid analogues. These FCS measurements of hybrid fluorescent tag–gold nanoparticle labeled lipids also revealed that commercially supplied streptavidin-coated gold nanoparticles contain large quantities of free streptavidin. Finally, the values of apparent diffusion coefficients obtained by STED-FCS and iSCAT differed by a factor of 2–3 across the techniques, while relative differences in mobility between different species of lipid analogues considered were identical in both approaches. In conclusion, our experiments reveal that large and potentially cross-linking scattering tags introduce a significant slow-down in diffusion on SLBs but no additional bias, and our labeling approach creates a new way of exploiting complementary information from STED-FCS and iSCAT measurements.

  17. A study of the dynamics of PTEN proteins in living cells using in vivo fluorescence correlation spectroscopy

    Du, Zhixue; Dong, Chaoqing; Ren, Jicun

    2017-06-01

    PTEN (phosphatase and tensin homolog on chromosome 10) is one of the most important tumor-suppressor proteins, which plays a key role in negative regulation of the PI3K/AKT pathway, and governs many cellular processes including growth, proliferation, survival and migration. The dynamics of PTEN proteins in single living cells is as yet unclear owing to a shortage of suitable in vivo approaches. Here, we report a single-molecule method for in vivo study of the dynamics of PTEN proteins in living cells using fluorescence correlation spectroscopy (FCS). First, we established a monoclonal H1299 stable cell line expressing enhanced green fluorescent protein (EGFP) and PTEN (EGFP-PTEN) fusion proteins; we then developed an in vivo FCS method to study the dynamics of EGFP-PTEN both in the nucleus and the cytoplasm. We investigated the diffusion behaviors of EGFP and EGFP-PTEN in solution, nucleus and cytosol, and observed that the motion of PTEN in living cells was restricted compared with EGFP. Finally, we investigated the protein dynamics in living cells under oxidative stress stimulation and a cellular ATP depletion treatment. Under oxidative stress stimulation, the EGFP-PTEN concentration increased in the nucleus, but slightly decreased in the cytoplasm. The diffusion coefficient and alpha value of EGFP-PTEN reduced significantly both in the nucleus and cytoplasm; the significantly decreased alpha parameter indicates a more restricted Brownian diffusion behavior. Under the cellular ATP depletion treatment, the concentration of EGFP-PTEN remained unchanged in the nucleus and decreased significantly in cytosol. The diffusion coefficient of EGFP-PTEN decreased significantly in cytosol, but showed no significant change in the nucleus; the alpha value decreased significantly in both the nucleus and cytoplasm. These results suggest that the concentration and mobility of PTEN in the nucleus and cytoplasm can be regulated by stimulation methods. Our approach provides a unique

  18. Wilson's disease: {sup 31}P and {sup 1}H MR spectroscopy and clinical correlation

    Sinha, Sanjib; Taly, A.B.; Prashanth, L.K. [National Institute of Mental Health and Neurosciences (NIMHANS), Department of Neurology, Bangalore (India); Ravishankar, S.; Vasudev, M.K. [National Institute of Mental Health and Neurosciences (NIMHANS), Department of Neuroimaging and Interventional Radiology, Bangalore (India)

    2010-11-15

    Proton ({sup 1}H) magnetic resonance spectroscopy (MRS) changes are noted in Wilson's disease (WD). However, there are no studies regarding membrane phospholipid abnormality using {sup 31}P MRS in these patients. We aimed to analyze the striatal spectroscopic abnormalities using {sup 31}P and {sup 1}H MRS in WD. Forty patients of WD (treated, 29; untreated,11) and 30 controls underwent routine MR image sequences and in vivo 2-D {sup 31}P and {sup 1}H MRS of basal ganglia using an image-selected technique on a 1.5-T MRI scanner. Statistical analysis was done using Student's t test. The mean durations of illness and treatment were 6.2 {+-} 7.4 and 4.8 {+-} 5.9 years, respectively. MRI images were abnormal in all the patients. {sup 1}H MRS revealed statistically significant reduction of N-acetyl aspartate (NAA)/choline (Cho) and NAA/creatine ratios in striatum ({sup 1}H MRS) of treated patients compared to controls. The mean values of phosphomonoesters (PME) (p < 0.0001), phosphodiesters (PDE) (p < 0.0001), and total phosphorus (TPh) (p < 0.0001) were elevated in patients compared to controls. Statistically significant elevated levels of ratio of PME/PDE (p = 0.05) observed in the striatum were noted in treated patients as compared to controls in the {sup 31}P MRS study. The duration of illness correlated well with increased PME/PDE [p < 0.001], PME/TPh [p < 0.05], and PDE/TPh [p < 0.05] and decreased NAA/Cho [p < 0.05] ratios. There was correlation of MRI score and reduced NAA/Cho ratio with disease severity. The PME/PDE ratio (right) was elevated in the treated group [p < 0.001] compared to untreated group. There is reduced breakdown and/or increased synthesis of membrane phospholipids and increased neuronal damage in basal ganglia in patients with WD. (orig.)

  19. Importance of Preserving Cross-correlation in developing Statistically Downscaled Climate Forcings and in estimating Land-surface Fluxes and States

    Das Bhowmik, R.; Arumugam, S.

    2015-12-01

    Multivariate downscaling techniques exhibited superiority over univariate regression schemes in terms of preserving cross-correlations between multiple variables- precipitation and temperature - from GCMs. This study focuses on two aspects: (a) develop an analytical solutions on estimating biases in cross-correlations from univariate downscaling approaches and (b) quantify the uncertainty in land-surface states and fluxes due to biases in cross-correlations in downscaled climate forcings. Both these aspects are evaluated using climate forcings available from both historical climate simulations and CMIP5 hindcasts over the entire US. The analytical solution basically relates the univariate regression parameters, co-efficient of determination of regression and the co-variance ratio between GCM and downscaled values. The analytical solutions are compared with the downscaled univariate forcings by choosing the desired p-value (Type-1 error) in preserving the observed cross-correlation. . For quantifying the impacts of biases on cross-correlation on estimating streamflow and groundwater, we corrupt the downscaled climate forcings with different cross-correlation structure.

  20. Multiscale modeling of interaction of alane clusters on Al(111) surfaces : a reactive force field and infrared absorbtion spectroscopy approach

    Ojwang, J.G.O.; Chaudhuri, S.; Duin, van A.C.T.; Chabal, Y.J.; Veyan, J.-F.; Santen, van R.A.; Kramer, G.J.; Goddard III, W.A.

    2010-01-01

    We have used reactive force field (ReaxFF) to investigate the mechanism of interaction of alanes on Al(111) surface. Our simulations show that, on the Al(111) surface, alanes oligomerize into larger alanes. In addition, from our simulations, adsorption of atomic hydrogen on Al(111) surface leads to

  1. Intracellular dynamics and fate of polystyrene nanoparticles in A549 Lung epithelial cells monitored by image (cross-) correlation spectroscopy and single particle tracking.

    Deville, Sarah; Penjweini, Rozhin; Smisdom, Nick; Notelaers, Kristof; Nelissen, Inge; Hooyberghs, Jef; Ameloot, Marcel

    2015-10-01

    Novel insights in nanoparticle (NP) uptake routes of cells, their intracellular trafficking and subcellular targeting can be obtained through the investigation of their temporal and spatial behavior. In this work, we present the application of image (cross-) correlation spectroscopy (IC(C)S) and single particle tracking (SPT) to monitor the intracellular dynamics of polystyrene (PS) NPs in the human lung carcinoma A549 cell line. The ensemble kinetic behavior of NPs inside the cell was characterized by temporal and spatiotemporal image correlation spectroscopy (TICS and STICS). Moreover, a more direct interpretation of the diffusion and flow detected in the NP motion was obtained by SPT by monitoring individual NPs. Both techniques demonstrate that the PS NP transport in A549 cells is mainly dependent on microtubule-assisted transport. By applying spatiotemporal image cross-correlation spectroscopy (STICCS), the correlated motions of NPs with the early endosomes, late endosomes and lysosomes are identified. PS NPs were equally distributed among the endolysosomal compartment during the time interval of the experiments. The cotransport of the NPs with the lysosomes is significantly larger compared to the other cell organelles. In the present study we show that the complementarity of ICS-based techniques and SPT enables a consistent elaborate model of the complex behavior of NPs inside biological systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Correlation between the structure and the piezoelectric properties of lead-free (K,Na,Li)(Nb,Ta,Sb)O3 ceramics studied by XRD and Raman spectroscopy.

    Rubio-Marcos, Fernando; Marchet, Pascal; Romero, Juan José; Fernández, Jose F

    2011-09-01

    This article reviews on the use of Raman spectroscopy for the study of (K,Na,Li)(Nb,Ta,Sb)O(3) lead-free piezoceramics. Currently, this material appears to be one of the most interesting and promising alternatives to the well-known PZT piezoelectric materials. In this work, we prepare piezoceramics with different stoichiometries and study their structural, ferroelectric, and piezoelectric properties. By using both Raman spectroscopy and X-ray diffraction, we establish a direct correlation between the structure and the properties. The results demonstrate that the wavenumber of the A(1g) vibration is proportional to the tetragonality, the remnant polarization, and the piezoelectric coefficients of these materials. Thus, Raman spectroscopy appears as a very useful technique for a fast evaluation of the crystalline structure and the ferroelectric/ piezoelectric properties.

  3. Fluid dynamic forces acting on a circular tube bundle in cross flow. Proposals of generation condition of vortex-induced vibration and correlation equation of turbulence-induced exciting force

    Inada, Fumio; Yoneda, Kimitoshi; Yasuo, Akira; Nishihara, Takashi

    2000-01-01

    In the circular tube bundle immersed in the crossflow, the exciting force induced by the turbulence and periodically discharged vortices becomes large, and it is necessary to confirm a long-term integrity to the flow induced vibration. In this report, the local fluid exciting force and the correlation length in the direction of tube axis were measured. The exciting force acting on the first row was smaller than that inside the tube bundle, and the exciting force was almost saturated at the third row. As for vortex induced vibration, there could be an influence when a dimensionless frequency was 0.4 or less. When vortex induced vibration did not affect the vibration, a correlation composed of a correlation length and power spectrum density of the local fluid exciting force were proposed, with which we could estimate the amplitude of the vibration. A computer program to estimate the vibration amplitude and maximum stress was made using the flow velocity distribution and the mode of vibration. (author)

  4. Anomalous random correlations of force constants on the lattice dynamical properties of disordered Au-Fe alloys

    Kangsabanik, Jiban; Chouhan, Rajiv K.; Johnson, D. D.; Alam, Aftab

    2017-09-01

    Gold iron (Au-Fe) alloys are of immense interest due to their biocompatibility, anomalous Hall conductivity, and applications in various medical treatments. However, irrespective of the method of preparation, they often exhibit a high level of disorder with properties sensitive to the thermal or magnetic annealing temperatures. We calculate the lattice dynamical properties of Au1 -xFex alloys using density functional theory methods where, being multisite properties, reliable interatomic force constant (IFC) calculations in disordered alloys remain a challenge. We follow a twofold approach: (1) an accurate IFC calculation in an environment with nominally zero chemical pair correlations to mimic the homogeneously disordered alloy and (2) a configurational averaging for the desired phonon properties (e.g., dispersion, density of states, and entropy). We find an anomalous change in the IFC's and phonon dispersion (split bands) near x =0.19 , which is attributed to the local stiffening of the Au-Au bonds when Au is in the vicinity of Fe. Other results based on mechanical and thermophysical properties reflect a similar anomaly: Phonon entropy, e.g., becomes negative below x =0.19 , suggesting a tendency for chemical unmixing, reflecting the onset of a miscibility gap in the phase diagram. Our results match fairly well with reported data wherever available.

  5. Water spray-induced grooming is negatively correlated with depressive behavior in the forced swimming test in rats.

    Shiota, Noboru; Narikiyo, Kimiya; Masuda, Akira; Aou, Shuji

    2016-05-01

    Rodents show grooming, a typical self-care behavior, under stress and non-stress conditions. Previous studies revealed that grooming under stress conditions such as the open-field test (OFT) or the elevated plus-maze test (EPM) is associated with anxiety, but the roles of grooming under non-stress conditions are not well understood. Here, we examined spray-induced grooming as a model of grooming under a non-stress condition to investigate the relationship between this grooming and depression-like behavior in the forced swim test (FST) and tail suspension test, and we compared spray-induced grooming with OFT- and EPM-induced grooming. The main finding was that the duration of spray-induced grooming, but not that of OFT/EPM-induced grooming, was negatively correlated with the duration of immobility in the FST, an index of depression-like behavior. The results suggest that spray-induced grooming is functionally different from the grooming in the OFT and EPM and is related to reduction of depressive behavior.

  6. Atomic force microscopy and Raman scattering spectroscopy studies on heat-induced fibrous aggregates of β-lactoglobulin

    Ikeda, Shinya

    2003-01-01

    Nanometer-thick fibrous aggregates of β-lactoglobulin alone and its mixture with other globular proteins were formed by heating aqueous solutions at pH 2 with maintaining an effective level of electrostatic repulsion among denatured protein molecules. In atomic force microscopy (AFM) images, these fibrous aggregates appeared to be fairly uniform in width and height and composed of strings of globular elements. Fibrous aggregates formed in β-lactoglobulin individual systems were only slightly ...

  7. Correlation between the local stress and the grain misorientation in the polycrystalline Al2O3 measured by near-field luminescence spectroscopy

    Tomimatsu, Toru; Takigawa, Ryo

    2018-06-01

    Owing to its high spatial resolution, near-field spectroscopy is a useful method for sensing the stress in a narrow region of submicron order. Here, on the basis of the highly resolved images obtained by near-field luminescence spectroscopy, we propose a statistical method of analyzing grain anisotropy-induced stress in polycrystalline Al2O3. We focus on two characteristics of a spectra: the intensity ratio and peak shift of luminescence of two lines (R1 and R2) from Al2O3 to discuss crystal orientation and stress, respectively. By incorporating the concept of the crystal misorientation parameter using intensity ratio, an apparent correlation between the magnitude of stress and the misorientation is found. This correlation analysis provides an important insight for the investigation of local thermal stress in Al2O3.

  8. Using optical fibers with different modes to improve the signal-to-noise ratio of diffuse correlation spectroscopy flow-oximeter measurements

    He, Lian; Lin, Yu; Shang, Yu; Shelton, Brent J.; Yu, Guoqiang

    2013-01-01

    The dual-wavelength diffuse correlation spectroscopy (DCS) flow-oximeter is an emerging technique enabling simultaneous measurements of blood flow and blood oxygenation changes in deep tissues. High signal-to-noise ratio (SNR) is crucial when applying DCS technologies in the study of human tissues where the detected signals are usually very weak. In this study, single-mode, few-mode, and multimode fibers are compared to explore the possibility of improving the SNR of DCS flow-oximeter measure...

  9. Source Determination of Red Gel Pen Inks using Raman Spectroscopy and Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy combined with Pearson's Product Moment Correlation Coefficients and Principal Component Analysis.

    Mohamad Asri, Muhammad Naeim; Mat Desa, Wan Nur Syuhaila; Ismail, Dzulkiflee

    2018-01-01

    The potential combination of two nondestructive techniques, that is, Raman spectroscopy (RS) and attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy with Pearson's product moment correlation (PPMC) coefficient (r) and principal component analysis (PCA) to determine the actual source of red gel pen ink used to write a simulated threatening note, was examined. Eighteen (18) red gel pens purchased from Japan and Malaysia from November to December 2014 where one of the pens was used to write a simulated threatening note were analyzed using RS and ATR-FTIR spectroscopy, respectively. The spectra of all the red gel pen inks including the ink deposited on the simulated threatening note gathered from the RS and ATR-FTIR analyses were subjected to PPMC coefficient (r) calculation and principal component analysis (PCA). The coefficients r = 0.9985 and r = 0.9912 for pairwise combination of RS and ATR-FTIR spectra respectively and similarities in terms of PC1 and PC2 scores of one of the inks to the ink deposited on the simulated threatening note substantiated the feasibility of combining RS and ATR-FTIR spectroscopy with PPMC coefficient (r) and PCA for successful source determination of red gel pen inks. The development of pigment spectral library had allowed the ink deposited on the threatening note to be identified as XSL Poppy Red (CI Pigment Red 112). © 2017 American Academy of Forensic Sciences.

  10. Membrane Disruption Mechanism of a Prion Peptide (106-126) Investigated by Atomic Force Microscopy, Raman and Electron Paramagnetic Resonance Spectroscopy.

    Pan, Jianjun; Sahoo, Prasana K; Dalzini, Annalisa; Hayati, Zahra; Aryal, Chinta M; Teng, Peng; Cai, Jianfeng; Rodriguez Gutierrez, Humberto; Song, Likai

    2017-05-18

    A fragment of the human prion protein spanning residues 106-126 (PrP106-126) recapitulates many essential properties of the disease-causing protein such as amyloidogenicity and cytotoxicity. PrP106-126 has an amphipathic characteristic that resembles many antimicrobial peptides (AMPs). Therefore, the toxic effect of PrP106-126 could arise from a direct association of monomeric peptides with the membrane matrix. Several experimental approaches are employed to scrutinize the impacts of monomeric PrP106-126 on model lipid membranes. Porous defects in planar bilayers are observed by using solution atomic force microscopy. Adding cholesterol does not impede defect formation. A force spectroscopy experiment shows that PrP106-126 reduces Young's modulus of planar lipid bilayers. We use Raman microspectroscopy to study the effect of PrP106-126 on lipid atomic vibrational dynamics. For phosphatidylcholine lipids, PrP106-126 disorders the intrachain conformation, while the interchain interaction is not altered; for phosphatidylethanolamine lipids, PrP106-126 increases the interchain interaction, while the intrachain conformational order remains similar. We explain the observed differences by considering different modes of peptide insertion. Finally, electron paramagnetic resonance spectroscopy shows that PrP106-126 progressively decreases the orientational order of lipid acyl chains in magnetically aligned bicelles. Together, our experimental data support the proposition that monomeric PrP106-126 can disrupt lipid membranes by using similar mechanisms found in AMPs.

  11. Water-induced morphology changes in an ultrathin silver film studied by ultraviolet-visible, surface-enhanced Raman scattering spectroscopy and atomic force microscopy

    Li Xiaoling; Xu Weiqing; Jia Huiying; Wang Xu; Zhao Bing; Li Bofu; Ozaki, Yukihiro

    2005-01-01

    Water-induced changes in the morphology and optical properties of an ultrathin Ag film (3 nm thickness) have been studied by use of ultraviolet-visible (UV-Vis) spectroscopy, atomic force microscopy (AFM) and surface-enhanced Raman scattering (SERS) spectroscopy. A confocal micrograph shows that infinite regular Ag rings with almost uniform size (4 μm) emerge on the film surface after the ultrathin Ag film was immersed into water. The AFM measurement further confirms that the Ag rings consist of some metal holes with pillared edges. The UV-Vis spectrum shows that an absorption band at 486 nm of the Ag film after the immersion in water (I-Ag film) blue shifts by 66 nm with a significant decrease in absorbance, which is attributed to the macroscopic loss of some Ag atoms and the change in the morphology of the Ag film. The polarized UV-Vis spectra show that a band at 421 nm due to the normal component of the plasmon oscillation blue shifts after immersing the ultrathin Ag film into water. This band is found to be strongly angle-dependent for p-polarized light, indicating that the optical properties of the ultrathin Ag film are changed. The I-Ag film is SERS-active, and the SERS enhancement depends on different active sites on the film surface. Furthermore, it seems that the orientation of an adsorbate is related to the morphology of the I-Ag film

  12. Phase transition behaviors of the supported DPPC bilayer investigated by sum frequency generation (SFG) vibrational spectroscopy and atomic force microscopy (AFM).

    Wu, Heng-Liang; Tong, Yujin; Peng, Qiling; Li, Na; Ye, Shen

    2016-01-21

    The phase transition behaviors of a supported bilayer of dipalmitoylphosphatidyl-choline (DPPC) have been systematically evaluated by in situ sum frequency generation (SFG) vibrational spectroscopy and atomic force microscopy (AFM). By using an asymmetric bilayer composed of per-deuterated and per-protonated monolayers, i.e., DPPC-d75/DPPC and a symmetric bilayer of DPPC/DPPC, we were able to probe the molecular structural changes during the phase transition process of the lipid bilayer by SFG spectroscopy. It was found that the DPPC bilayer is sequentially melted from the top (adjacent to the solution) to bottom leaflet (adjacent to the substrate) over a wide temperature range. The conformational ordering of the supported bilayer does not decrease (even slightly increases) during the phase transition process. The conformational defects in the bilayer can be removed after the complete melting process. The phase transition enthalpy for the bottom leaflet was found to be approximately three times greater than that for the top leaflet, indicating a strong interaction of the lipids with the substrate. The present SFG and AFM observations revealed similar temperature dependent profiles. Based on these results, the temperature-induced structural changes in the supported lipid bilayer during its phase transition process are discussed in comparison with previous studies.

  13. Revealing energy level structure of individual quantum dots by tunneling rate measured by single-electron sensitive electrostatic force spectroscopy.

    Roy-Gobeil, Antoine; Miyahara, Yoichi; Grutter, Peter

    2015-04-08

    We present theoretical and experimental studies of the effect of the density of states of a quantum dot (QD) on the rate of single-electron tunneling that can be directly measured by electrostatic force microscopy (e-EFM) experiments. In e-EFM, the motion of a biased atomic force microscope cantilever tip modulates the charge state of a QD in the Coulomb blockade regime. The charge dynamics of the dot, which is detected through its back-action on the capacitavely coupled cantilever, depends on the tunneling rate of the QD to a back-electrode. The density of states of the QD can therefore be measured through its effect on the energy dependence of tunneling rate. We present experimental data on individual 5 nm colloidal gold nanoparticles that exhibit a near continuous density of state at 77 K. In contrast, our analysis of already published data on self-assembled InAs QDs at 4 K clearly reveals discrete degenerate energy levels.

  14. Extended x-ray absorption fine structure spectroscopy and x-ray absorption near edge spectroscopy study of aliovalent doped ceria to correlate local structural changes with oxygen vacancies clustering

    Shirbhate, S. C.; Acharya, S. A., E-mail: saha275@yahoo.com [Department of Physics, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur 440033 (India); Yadav, A. K. [Atomic and molecular Physics Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2016-04-04

    This study provides atomic scale insight to understand the role of aliovalent dopants on oxygen vacancies clustering and dissociation mechanism in ceria system in order to enhance the performance of oxy-ion conductor. Dopants induced microscale changes in ceria are probed by extended X-ray absorption fine structure spectroscopy, X-ray absorption near edge spectra, and Raman spectroscopy. The results are explored to establish a correlation between atomic level structural changes (coordination number, interatomic spacing) → formation of dimer and trimer type cation-oxygen vacancies defect complex (intrinsic and extrinsic) → dissociation of oxygen vacancies from defect cluster → ionic conductivity temperature. It is a strategic approach to understand key physics of ionic conductivity mechanism in order to reduce operating temperature of electrolytes for intermediate temperature (300–450 °C) electrochemical devices for the first time.

  15. Atomic force microscopy and X-ray photoelectron spectroscopy study of NO2 reactions on CaCO3 (1014) surfaces in humid environments.

    Baltrusaitis, Jonas; Grassian, Vicki H

    2012-09-13

    In this study, alternating current (AC) mode atomic force microscopy (AFM) combined with phase imaging and X-ray photoelectron spectroscopy (XPS) were used to investigate the effect of nitrogen dioxide (NO2) adsorption on calcium carbonate (CaCO3) (101̅4) surfaces at 296 K in the presence of relative humidity (RH). At 70% RH, CaCO3 (101̅4) surfaces undergo rapid formation of a metastable amorphous calcium carbonate layer, which in turn serves as a substrate for recrystallization of a nonhydrated calcite phase, presumably vaterite. The adsorption of nitrogen dioxide changes the surface properties of CaCO3 (101̅4) and the mechanism for formation of new phases. In particular, the first calcite nucleation layer serves as a source of material for further island growth; when it is depleted, there is no change in total volume of nitrocalcite, Ca(NO3)2, particles formed whereas the total number of particles decreases. This indicates that these particles are mobile and coalesce. Phase imaging combined with force curve measurements reveals areas of inhomogeneous energy dissipation during the process of water adsorption in relative humidity experiments, as well as during nitrocalcite particle formation. Potential origins of the different energy dissipation modes within the sample are discussed. Finally, XPS analysis confirms that NO2 adsorbs on CaCO3 (101̅4) in the form of nitrate (NO3(-)) regardless of environmental conditions or the pretreatment of the calcite surface at different relative humidity.

  16. Decoding Nucleation and Growth of Zeolitic Imidazolate Framework Thin Films with Atomic Force Microscopy and Vibrational Spectroscopy.

    Öztürk, Zafer; Filez, Matthias; Weckhuysen, Bert M

    2017-08-10

    The synthesis of metal-organic framework (MOF) thin films has garnered significant attention during the past decade. By better understanding the parameters governing the nucleation and growth of such thin films, their properties can be rationally tuned, empowering their application as (reactive) membranes. Here, a combined AFM-vibrational spectroscopy research strategy is employed to detail the chemistries governing the nucleation and growth of zeolitic imidazolate framework (ZIF) thin films, in particular isostructural Co-ZIF-67 and Zn-ZIF-8. First, a single step direct synthesis approach is used to investigate the influence of different synthesis parameters -metal/linker ratio, temperature, and metal type- on the thin film nucleation and growth behaviour. While the metal/linker ratio has a pronounced effect on the thin film nucleation rate, the temperature mainly influences the growth kinetics of nuclei forming the thin film. In addition, the nucleation and growth of ZIF thin films is shown to be highly dependent on the electronegativity of the metal type. Thin-film thickness control can be achieved by using a multistep synthesis strategy, implying repetitive applications of single step deposition under identical synthesis conditions, for which a growth mechanism is proposed. This study provides insight into the influence of synthesis parameters on the ZIF thin film properties, using tools at hand to rationally tune MOF thin film properties. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  17. Analytical Raman spectroscopic study for discriminant analysis of different animal-derived feedstuff: Understanding the high correlation between Raman spectroscopy and lipid characteristics.

    Gao, Fei; Xu, Lingzhi; Zhang, Yuejing; Yang, Zengling; Han, Lujia; Liu, Xian

    2018-02-01

    The objectives of the current study were to explore the correlation between Raman spectroscopy and lipid characteristics and to assess the potential of Raman spectroscopic methods for distinguishing the different sources of animal-originated feed based on lipid characteristics. A total of 105 lipid samples derived from five animal species have been analyzed by gas chromatography (GC) and FT-Raman spectroscopy. High correlations (r 2 >0.94) were found between the characteristic peak ratio of the Raman spectra (1654/1748 and 1654/1445) and the degree of unsaturation of the animal lipids. The results of FT-Raman data combined with chemometrics showed that the fishmeal, poultry, porcine and ruminant (bovine and ovine) MBMs could be well separated based on their lipid spectral characteristics. This study demonstrated that FT-Raman spectroscopy can mostly exhibit the lipid structure specificity of different species of animal-originated feed and can be used to discriminate different animal-originated feed samples. Copyright © 2017. Published by Elsevier Ltd.

  18. Single molecule force spectroscopy for in-situ probing oridonin inhibited ROS-mediated EGF-EGFR interactions in living KYSE-150 cells.

    Pi, Jiang; Jin, Hua; Jiang, Jinhuan; Yang, Fen; Cai, Huaihong; Yang, Peihui; Cai, Jiye; Chen, Zheng W

    2017-05-01

    As the active anticancer component of Rabdosia Rubescens, oridonin has been proved to show strong anticancer activity in cancer cells, which is also found to be closely related to its specific inhibition effects on the EGFR tyrosine kinase activity. In this study, atomic force microscopy based single molecule force spectroscopy (AFM-SMFS) was used for real-time and in-situ detection of EGF-EGFR interactions in living esophageal cancer KYSE-150 cells to evaluate the anticancer activity of oridonin for the first time. Oridonin was found to induce apoptosis and also reduce EGFR expression in KYSE-150 cells. AFM-SMFS results demonstrated that oridonin could inhibit the binding between EGF and EGFR in KYSE-150 cells by decreasing the unbinding force and binding probability for EGF-EGFR complexes, which was further proved to be closely associated with the intracellular ROS level. More precise mechanism studies based on AFM-SMFS demonstrated that oridonin treatment could decrease the energy barrier width, increase the dissociation off rate constant and decrease the activation energy of EGF-EGFR complexes in ROS dependent way, suggesting oridonin as a strong anticancer agent targeting EGF-EGFR interactions in cancer cells through ROS dependent mechanism. Our results not only suggested oridonin as a strong anticancer agent targeting EGF-EGFR interactions in ROS dependent mechanism, but also highlighted AFM-SMFS as a powerful technique for pharmacodynamic studies by detecting ligand-receptor interactions, which was also expected to be developed into a promising tool for the screening and mechanism studies of drugs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Interactions of Histone Acetyltransferase p300 with the Nuclear Proteins Histone and HMGB1, As Revealed by Single Molecule Atomic Force Spectroscopy.

    Banerjee, S; Rakshit, T; Sett, S; Mukhopadhyay, R

    2015-10-22

    One of the important properties of the transcriptional coactivator p300 is histone acetyltransferase (HAT) activity that enables p300 to influence chromatin action via histone modulation. p300 can exert its HAT action upon the other nuclear proteins too--one notable example being the transcription-factor-like protein HMGB1, which functions also as a cytokine, and whose accumulation in the cytoplasm, as a response to tissue damage, is triggered by its acetylation. Hitherto, no information on the structure and stability of the complexes between full-length p300 (p300FL) (300 kDa) and the histone/HMGB1 proteins are available, probably due to the presence of unstructured regions within p300FL that makes it difficult to be crystallized. Herein, we have adopted the high-resolution atomic force microscopy (AFM) approach, which allows molecularly resolved three-dimensional contour mapping of a protein molecule of any size and structure. From the off-rate and activation barrier values, obtained using single molecule dynamic force spectroscopy, the biochemical proposition of preferential binding of p300FL to histone H3, compared to the octameric histone, can be validated. Importantly, from the energy landscape of the dissociation events, a model for the p300-histone and the p300-HMGB1 dynamic complexes that HAT forms, can be proposed. The lower unbinding forces of the complexes observed in acetylating conditions, compared to those observed in non-acetylating conditions, indicate that upon acetylation, p300 tends to weakly associate, probably as an outcome of charge alterations on the histone/HMGB1 surface and/or acetylation-induced conformational changes. To our knowledge, for the first time, a single molecule level treatment of the interactions of HAT, where the full-length protein is considered, is being reported.

  20. Study of optoelectronic properties of thin film solar cell materials Cu2ZnSn(S,Se)4 using multiple correlative spatially-resolved spectroscopy techniques

    Chen, Qiong

    Containing only earth abundant and environmental friendly elements, quaternary compounds Cu2ZnSnS4 (CZTS) and Cu2ZnSnSe 4 (CZTSe) are considered as promising absorber materials for thin film solar cells. The best record efficiency for this type of thin film solar cell is now 12.6%. As a promising photovoltaic (PV) material, the electrical and optical properties of CZTS(Se) have not been well studied. In this work, an effort has been made to understand the optoelectronic and structural properties, in particular the spatial variations, of CZTS(Se) materials and devices by correlating multiple spatially resolved characterization techniques with sub-micron resolution. Micro-Raman (micro-Raman) spectroscopy was used to analyze the chemistry compositions in CZTS(Se) film; Micro-Photoluminescence (micro-PL) was used to determine the band gap and possible defects. Micro-Laser-Beam-Induced-Current (micro-LBIC) was used to examine the photo-response of CZTS(Se) solar cell in different illumination conditions. Micro-reflectance was used to estimate the reflectance loss. And Micro-I-V measurement was used to compare important electrical parameters from CZTS(Se) solar cells with different device structure or absorber compositions. Scanning electron microscopy and atomic force microscopy were used to characterize the surface morphology. Successfully integrating and correlating these techniques was first demonstrated during the course of this work in our laboratory, and this level of integration and correlation has been rare in the field of PV research. This effort is significant not only for this particular project and also for a wide range of research topics. Applying this approach, in conjunction with high-temperature and high-excitation-power optical spectroscopy, we have been able to reveal the microscopic scale variations among samples and devices that appeared to be very similar from macroscopic material and device characterizations, and thus serve as a very powerful tool

  1. A correlative study on data from pork carcass and processed meat (Bauernspeck) for automatic estimation of chemical parameters by means of near-infrared spectroscopy.

    Boschetti, Lucio; Ottavian, Matteo; Facco, Pierantonio; Barolo, Massimiliano; Serva, Lorenzo; Balzan, Stefania; Novelli, Enrico

    2013-11-01

    The use of near-infrared spectroscopy (NIRS) is proposed in this study for the characterization of the quality parameters of a smoked and dry-cured meat product known as Bauernspeck (originally from Northern Italy), as well as of some technological traits of the pork carcass used for its manufacturing. In particular, NIRS is shown to successfully estimate several key quality parameters (including water activity, moisture, dry matter, ash and protein content), suggesting its suitability for real time application in replacement of expensive and time consuming chemical analysis. Furthermore, a correlative approach based on canonical correlation analysis was used to investigate the spectral regions that are mostly correlated to the characteristics of interest. The identification of these regions, which can be linked to the absorbance of the main functional chemical groups, is intended to provide a better understanding of the chemical structure of the substrate under investigation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Characterization of the molecular structure and mechanical properties of polymer surfaces and protein/polymer interfaces by sum frequency generation vibrational spectroscopy and atomic force microscopy

    Koffas, Telly Stelianos [Univ. of California, Berkeley, CA (United States)

    2004-01-01

    Sum frequency generation (SFG) vibrational spectroscopy, atomic force microscopy (AFM), and other complementary surface-sensitive techniques have been used to study the surface molecular structure and surface mechanical behavior of biologically-relevant polymer systems. SFG and AFM have emerged as powerful analytical tools to deduce structure/property relationships, in situ, for polymers at air, liquid and solid interfaces. The experiments described in this dissertation have been performed to understand how polymer surface properties are linked to polymer bulk composition, substrate hydrophobicity, changes in the ambient environment (e.g., humidity and temperature), or the adsorption of macromolecules. The correlation of spectroscopic and mechanical data by SFG and AFM can become a powerful methodology to study and engineer materials with tailored surface properties. The overarching theme of this research is the interrogation of systems of increasing structural complexity, which allows us to extend conclusions made on simpler model systems. We begin by systematically describing the surface molecular composition and mechanical properties of polymers, copolymers, and blends having simple linear architectures. Subsequent chapters focus on networked hydrogel materials used as soft contact lenses and the adsorption of protein and surfactant at the polymer/liquid interface. The power of SFG is immediately demonstrated in experiments which identify the chemical parameters that influence the molecular composition and ordering of a polymer chain's side groups at the polymer/air and polymer/liquid interfaces. In general, side groups with increasingly greater hydrophobic character will be more surface active in air. Larger side groups impose steric restrictions, thus they will tend to be more randomly ordered than smaller hydrophobic groups. If exposed to a hydrophilic environment, such as water, the polymer chain will attempt to orient more of its hydrophilic groups to

  3. Recoil ion momentum spectroscopy in atomic and nuclear physics: applications to low energy ion-atom/molecule collisions and to beta-neutrino angular correlation in beta decay

    Flechard, X.

    2012-12-01

    Since the early 1990's, Recoil Ion Momentum Spectroscopy is an ideal tool for ion-atom and ion-molecule collisions study. We detail here the development of this experimental technique during the last twenty years, illustrated with some of the most striking results obtained at GANIL (Caen) and J.R. Mac Donald Laboratory (Kansas State University). Recoil Ion Momentum Spectroscopy is also particularly well suited for β-ν angular correlation measurements in nuclear β decay. The LPCTrap experiment, installed at GANIL, is based on this technique, coupled to the use of a Paul trap for the radioactive ions confinement. The precise measurements performed with this setup allow both, to test specific aspects of the Standard Model of elementary particles, and to study the electron shake-off process following β decay. (author)

  4. Correlates of Forced Sex Among Young Men Who Have Sex With Men in Yangon and Monywa, Myanmar.

    Johnston, Lisa G; Mon, Myo Myo; Steinhaus, Mara; Sass, Justine

    2017-05-01

    Forced sex at an early age is associated with a variety of negative factors including increased illness, high-risk sexual and substance-use behaviors, and mental and psychological stress. These sequelae may be compounded for men who have sex with men (MSM), especially young MSM and those with feminine gender identity and expression. This survey examined the prevalence and associations of forced sex among young MSM in two cities in Myanmar. In 2013-2014, surveys using respondent-driven sampling collected data on 200 young MSM in Yangon and 200 in Monywa. One quarter of young MSM in Yangon and 21 % in Monywa reported ever experiencing forced sex. In a multivariable model, having problems with family members and having any MSM friends with many partners had higher odds of experiencing forced sex. Having maternal acceptance of same-sex attraction (compared to acceptance by both parents) and becoming aware of their same-sex attraction at or above the age of 16 had lower odds of experiencing forced sex. Focused research is needed to understand the family and other social dynamics affecting vulnerability to forced sex, as well as specific sexual risks associated with forced sex among young MSM, including HIV acquisition and transmission risks.

  5. Occlusal force is correlated with cognitive function directly as well as indirectly via food intake in community-dwelling older Japanese: From the SONIC study.

    Ikebe, Kazunori; Gondo, Yasuyuki; Kamide, Kei; Masui, Yukie; Ishizaki, Taturo; Arai, Yasumichi; Inagaki, Hiroki; Nakagawa, Takeshi; Kabayama, Mai; Ryuno, Hirochika; Okubo, Hitomi; Takeshita, Hajime; Inomata, Chisato; Kurushima, Yuko; Mihara, Yusuke; Hatta, Kohdai; Fukutake, Motoyoshi; Enoki, Kaori; Ogawa, Taiji; Matsuda, Ken-Ichi; Sugimoto, Ken; Oguro, Ryosuke; Takami, Yoichi; Itoh, Norihisa; Takeya, Yasushi; Yamamoto, Koichi; Rakugi, Hiromi; Murakami, Shinya; Kitamura, Masahiro; Maeda, Yoshinobu

    2018-01-01

    Growing evidence suggests that oral health may be an important factor associated with cognitive function in aged populations. However, many previous studies on this topic used insensitive oral indicators or did not include certain essential covariates. Thus, we examined the association between occlusal force and cognitive function in a large sample of older adults, controlling for dietary intake, vascular risk factors, inflammatory biomarkers, depression, and genetic factors. In this cross-sectional study of older community-dwelling Japanese adults, we examined data collected from 994 persons aged 70 years and 968 persons aged 80 years. Cognitive function was measured using the Japanese version of the Montreal Cognitive Assessment (MoCA-J). Oral status and function were evaluated according to the number of remaining teeth, periodontal pocket depth, and maximal occlusal force. Associations between MoCA-J scores and occlusal force were investigated via bivariate and multivariate analyses. Education level, financial status, depression score, and intake of green and yellow vegetables, as well as number of teeth and occlusal force, were significantly correlated with MoCA-J scores in both age groups. Among individuals aged 80 years, CRP and periodontal status were weakly but significantly associated with MoCA-J score. After controlling for all significant variables via bivariate analyses, the correlation between maximal occlusal force and cognitive function persisted. A path analysis confirmed the hypothesis that cognitive function is associated with occlusal force directly as well as indirectly via food intake. After controlling for possible factors, maximal occlusal force was positively associated with cognitive function directly as well as indirectly through dietary intake.

  6. Direct correlation of charge transfer absorption with molecular donor:acceptor interfacial area via photothermal deflection spectroscopy

    Domingo, Ester

    2015-04-09

    We show that the Charge Transfer (CT) absorption signal in bulk-heterojunction (BHJ) solar cell blends, measured by photothermal deflection spectroscopy (PDS), is directly proportional to the density of molecular donor/acceptor interfaces. Since the optical transitions from ground state to the interfacial CT state are weakly allowed at photon energies below the optical gap of both donor and acceptor, we can exploit the use of this sensitive linear absorption spectroscopy for such quantification. Moreover, we determine the absolute molar extinction coefficient of the CT transition for an archetypical polymer-fullerene interface. The latter is ~100 times lower than the extinction coefficient of the donor chromophore involved, allowing us to experimentally estimate the transition dipole moment (0.3 D) and the electronic coupling between ground state and CT state to be on the order of 30 meV.

  7. Correlation analysis between surface electromyography and continuous-wave near-infrared spectroscopy parameters during isometric exercise to volitional fatigue

    ŞAYLİ, Ömer; AKIN, Ata; ÇOTUK, Hasan Birol

    2014-01-01

    In this study, the process of muscular fatigue was examined using surface electromyography (sEMG) and continuous-wave near-infrared spectroscopy (cw-NIRS) simultaneously during an isometric hand grip exercise at 50% and 75% of the maximal voluntary contraction (MVC), sustained until volitional fatigue. The mean frequency of the sEMG decreased during the whole exercise, whereas the root mean square had a tendency to increase. Oxyhemoglobin/deoxyhemoglobin concentration changes computed ...

  8. Two-trace two-dimensional (2T2D) correlation spectroscopy - A method for extracting useful information from a pair of spectra

    Noda, Isao

    2018-05-01

    Two-trace two-dimensional (2T2D) correlation spectroscopy, where a pair of spectra are compared as 2D maps by a form of cross correlation analysis, is introduced. In 2T2D, spectral intensity changes of bands arising from the same origin, which cannot change independently of each other, are synchronized. Meanwhile, those arising from different sources may and often do change asynchronously. By taking advantage of this property, one can distinguish and classify a number of contributing bands present in the original pair of spectra in a systematic manner. Highly overlapped neighboring bands originating from different sources can also be identified by the presence of asynchronous cross peaks, thus enhancing the apparent spectral resolution. Computational procedure to obtain 2T2D correlation spectra and their interpretation method, as well as an illustrative description of the basic concept in the vector phase space, are provided. 2T2D spectra may also be viewed as individual building blocks of the generalized 2D correlation spectra derived from a series of more than two spectral data. Some promising application potentials of 2T2D correlation and integration with established advanced 2D correlation techniques are discussed.

  9. Hippocampal proton MR spectroscopy as a novel approach in the assessment of radiation injury and the correlation to neurocognitive function impairment: initial experiences

    Pospisil, Petr; Kazda, Tomas; Bulik, Martin; Dobiaskova, Marie; Burkon, Petr; Hynkova, Ludmila; Slampa, Pavel; Jancalek, Radim

    2015-01-01

    The hippocampus is considered as the main radiosensitive brain structure responsible for postradiotherapy cognitive decline. We prospectively assessed correlation of memory change to hippocampal N-acetylaspartate (h-tNAA) concentration, a neuronal density and viability marker, by 1 H-MR spectroscopy focused on the hippocampus. Patients with brain metastases underwent whole brain radiotherapy (WBRT) to a dose of 30 Gy in ten fractions daily. Pre-radiotherapy 1 H-MR spectroscopy focused on the h-tNAA concentration and memory testing was performed. Memory was evaluated by Auditory Verbal Learning Test (AVLT) and Brief Visuospatial Memory Test-Revised (BVMT-R). Total recall, recognition and delayed recall were reported. The both investigation procedures were repeated 4 months after WBRT and the h-tNAA and memory changes were correlated. Of the 20 patients, ten passed whole protocol. The h-tNAA concentration significantly decreased from pre-WBRT 8.9, 8.86 and 8.88 [mM] in the right, left and both hippocampi to 7.16, 7.65 and 7.4 after WBRT, respectively. In the memory tests a significant decrease was observed in AVLT total-recall, BVMT-R total-recall and BVMT-R delayed-recall. Weak to moderate correlations were observed between left h-tNAA and AVLT recognition and all BVMT-R subtests and between the right h-tNAA and AVLT total-recall. A significant decrease in h-tNAA after WBRT was proven by 1 H-MR spectroscopy as a feasible method for the in vivo investigation of radiation injury. Continuing patient recruitment focusing on other cognitive tests and metabolites is needed

  10. Chemical transitions of Areca semen during the thermal processing revealed by temperature-resolved ATR-FTIR spectroscopy and two-dimensional correlation analysis

    Wang, Zhibiao; Wang, Xu; Pei, Wenxuan; Li, Sen; Sun, Suqin; Zhou, Qun; Chen, Jianbo

    2018-03-01

    Areca semen is a common herb used in traditional Chinese medicine, but alkaloids in this herb are categorized as Group I carcinogens by IARC. It has been proven that the stir-baking process can reduce alkaloids in Areca semen while keep the activity for promoting digestion. However, the changes of compositions other than alkaloids during the thermal processing are unclear. Understanding the thermal chemical transitions of Areca semen is necessary to explore the processing mechanisms and optimize the procedures. In this research, FTIR spectroscopy with a temperature-controlled ATR accessory is employed to study the heating process of Areca semen. Principal component analysis and two-dimensional correlation spectroscopy are used to interpret the spectra to reveal the chemical transitions of Areca semen in different temperature ranges. The loss of a few volatile compounds in the testa and sperm happens below 105 °C, while some esters in the sperm decreases above 105 °C. As the heating temperature is close to 210 °C, Areca semen begins to be scorched and the decomposition of many compounds can be observed. This research shows the potential of the temperature-resolved ATR-FTIR spectroscopy in exploring the chemical transitions of the thermal processing of herbal materials.

  11. Investigation of the Brill transition in nylon 6,6 by Raman, THz-Raman, and two-dimensional correlation spectroscopy.

    Bertoldo Menezes, D; Reyer, A; Musso, M

    2018-02-05

    The Brill transition is a phase transition process in polyamides related with structural changes between the hydrogen bonds of the lateral functional groups (CO) and (NH). In this study, we have used the potential of Raman spectroscopy for exploring this phase transition in polyamide 6,6 (nylon 6,6), due to the sensitivity of this spectroscopic technique to small intermolecular changes affecting vibrational properties of relevant functional groups. During a step by step heating and cooling process of the sample we collected Raman spectra allowing us from two-dimensional Raman correlation spectroscopy to identify which spectral regions suffered the largest influence during the Brill transition, and from Terahertz Stokes and anti-Stokes Raman spectroscopy to obtain complementary information, e.g. on the temperature of the sample. This allowed us to grasp signatures of the Brill transition from peak parameters of vibrational modes associated with (CC) skeletal stretches and (CNH) bending, and to verify the Brill transition temperature at around 160°C, as well as the reversibility of this phase transition. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Two-dimensional correlation spectroscopy reveals the underlying compositions for FT-NIR identification of the medicinal bulbs of the genus Fritillaria

    Chen, Jianbo; Wang, Yue; Liu, Aoxue; Rong, Lixin; Wang, Jingjuan

    2018-03-01

    Fritillariae Bulbus, the dried bulbs of several species of the genus Fritillaria, is often used in traditional Chinese medicine for the treatment of cough and pulmonary diseases. However, the similar appearances make it difficult to identify different kinds of Fritillariae Bulbus. In this research, Fourier transform near-infrared (FT-NIR) spectroscopy with a reflection fiber probe is employed for the direct testing and automatic identification of different kinds of Fritillariae Bulbus to ensure the authenticity, efficacy and safety. The bulbs can be measured directly without pulverizing. According to the two-dimensional (2D) correlation analysis and statistical analysis, the height ratio of the two peaks near 4860 cm-1 and 4750 cm-1 in the second derivative spectra is specific to the species of Fritillariae Bulbus. This indicates that the relative amount of protein and carbohydrate may be critical to identify Fritillariae Bulbus. With the help of the SIMCA model, the four kinds of Fritillariae Bulbus can be identified correctly by FT-NIR spectroscopy. The results show the potential of FT-NIR spectroscopy with a reflection fiber probe in the rapid testing and identification of Fritillariae Bulbus.

  13. Ultrahigh-Resolution Magnetic Resonance in Inhomogeneous Magnetic Fields: Two-Dimensional Long-Lived-Coherence Correlation Spectroscopy

    Chinthalapalli, Srinivas; Bornet, Aurélien; Segawa, Takuya F.; Sarkar, Riddhiman; Jannin, Sami; Bodenhausen, Geoffrey

    2012-07-01

    A half-century quest for improving resolution in Nuclear Magnetic Resonance (NMR) and Magnetic Resonance Imaging (MRI) has enabled the study of molecular structures, biological interactions, and fine details of anatomy. This progress largely relied on the advent of sophisticated superconducting magnets that can provide stable and homogeneous fields with temporal and spatial variations below ΔB0/B0LLC-COSY) opens the way to overcome both inhomogeneous and homogeneous broadening, which arise from local variations in static fields and fluctuating dipole-dipole interactions, respectively. LLC-COSY makes it possible to obtain ultrahigh resolution two-dimensional spectra, with linewidths on the order of Δν=0.1 to 1 Hz, even in very inhomogeneous fields (ΔB0/B0>10ppm or 5000 Hz at 9.7 T), and can improve resolution by a factor up to 9 when the homogeneous linewidths are determined by dipole-dipole interactions. The resulting LLC-COSY spectra display chemical shift differences and scalar couplings in two orthogonal dimensions, like in “J spectroscopy.” LLC-COSY does not require any sophisticated gradient switching or frequency-modulated pulses. Applications to in-cell NMR and to magnetic resonance spectroscopy (MRS) of selected volume elements in MRI appear promising, particularly when susceptibility variations tend to preclude high resolution.

  14. Chemical analysis of industrial scale deposits by combined use of correlation coefficients with emission line detection of laser induced breakdown spectroscopy spectra

    Siozos, P.; Philippidis, A.; Hadjistefanou, M.; Gounarakis, C.; Anglos, D.

    2013-01-01

    Laser-induced breakdown spectroscopy (LIBS) was used to determine the mineral composition of various industrial scale samples. The aim of the study has been to investigate the capacity of LIBS to provide a fast, reliable analytical tool for carrying out routine analysis of inorganic scales, potentially on site, as a means to facilitate decision making concerning scale removal procedures. LIBS spectra collected in the range of 200–660 nm conveyed information about the metal content of the minerals. Via a straightforward analysis based on linear correlation of LIBS spectra it was possible to successfully discriminate scale samples into three main groups, Fe-rich, Ca-rich and Ba-rich, on the basis of correlation coefficients. By combining correlation coefficients with spectral data collected in the NIR, 860–960 nm, where sulfur emissions are detected, it became further possible to discriminate sulfates from carbonates as confirmed by independent analysis based on Raman spectroscopy. It is emphasized that the proposed LIBS-based method successfully identifies the major mineral or minerals present in the samples classifying the scales into relevant groups hence enabling process engineers to select appropriate scale dissolution strategies. - Highlights: • LIBS was used to determine the mineral composition of industrial scale samples. • Three groups of inorganic scales were identified: Ca rich, Ba rich and Fe rich. • A method that combines correlation coefficients and line detection is proposed. • The method successfully identifies the main mineral, or minerals, in the samples. • The results were compared with results obtained by use of Raman analysis

  15. Corrosive effects of fluoride on titanium: investigation by X-ray photoelectron spectroscopy, atomic force microscopy, and human epithelial cell culturing.

    Stájer, Anette; Ungvári, Krisztina; Pelsoczi, István K; Polyánka, Hilda; Oszkó, Albert; Mihalik, Erzsébet; Rakonczay, Zoltán; Radnai, Márta; Kemény, Lajos; Fazekas, András; Turzó, Kinga

    2008-11-01

    High fluoride (F(-)) concentrations and acidic pH impair the corrosion resistance of titanium (Ti). Effects of F(-)-containing caries-preventive prophylactic rinses, and gels on Ti were investigated by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Human epithelial cell attachment and proliferation were investigated by dimethylthiazol-diphenyl tetrazolium bromide (MTT) and protein content assays. Aqueous 1% NaF solution (3800 ppm F(-), pH 4.5) or high (12,500 ppm) F(-) content gel (pH 4.8) strongly corroded the surface and modified its composition. XPS revealed formation of a strongly bound F(-)-containing complex (Na(2)TiF(6)). AFM indicated an increase in roughness (R(a)) of the surfaces: 10-fold for the NaF solution and smaller for the gel or a mouthwash (250 ppm F(-), pH 4.4). MTT revealed that cell attachment was significantly increased by the gel, but was not disturbed by either the mouthwash or the NaF. Cell proliferation determined by MTT decreased significantly only for the NaF-treated samples; protein content assay experiments showed no such effect. This study indicates that epithelial cell culturing results can depend on the method used, and the adverse effects of a high F(-) concentration and low pH should be considered when prophylactic gels are applied by patients with Ti implants or other dental devices.

  16. Fragmentation of the C60 molecule in collision with light ions studied by a multi-correlation technique. Cross-sections, electron spectroscopy

    Rentenier, A.

    2004-04-01

    A quantitative study of the C60 fullerenes fragmentation in collision with light ions (H n + with n=1,2,3, He q+ with q=1,2) in the velocity range 0,1 - 2,3 u.a.) is presented. The multi-correlation technique, developed between fragment ions and electrons with well defined energy, has enlightened some of the dependences and properties of fragmentation mechanisms (cross sections, electron spectroscopy, size distributions, kinetic energy of fragment ions, Campi's scatter plot, activation energies). The deposited energy hence appeared as an important parameter. Cross sections have been measured, for the first time, for all the collisional processes. Ionisation and capture only depends on the collision velocity. On the other hand, scaling laws with the deposited energy have been observed for the cross sections of multifragmentation, which depends on the collision energy and the nature of the projectile. The deposited energy has also been found as an essential parameter to understand the evolution of the charged fragment size distributions. The electron spectroscopy, achieved at an emission angle of 35 degrees, showed spectra peaked at important energies (from 5 to 20 eV). The spectra shape depends on the collision velocity. A first theoretical analysis points out the link between the observed energy distribution and the presence of a centrifugal potential barrier. Finally, correlation experiments between produced ions and electron energy reveal that electron energy increases with internal energy. (author)

  17. A Rapid Identification Method for Calamine Using Near-Infrared Spectroscopy Based on Multi-Reference Correlation Coefficient Method and Back Propagation Artificial Neural Network.

    Sun, Yangbo; Chen, Long; Huang, Bisheng; Chen, Keli

    2017-07-01

    As a mineral, the traditional Chinese medicine calamine has a similar shape to many other minerals. Investigations of commercially available calamine samples have shown that there are many fake and inferior calamine goods sold on the market. The conventional identification method for calamine is complicated, therefore as a result of the large scale of calamine samples, a rapid identification method is needed. To establish a qualitative model using near-infrared (NIR) spectroscopy for rapid identification of various calamine samples, large quantities of calamine samples including crude products, counterfeits and processed products were collected and correctly identified using the physicochemical and powder X-ray diffraction method. The NIR spectroscopy method was used to analyze these samples by combining the multi-reference correlation coefficient (MRCC) method and the error back propagation artificial neural network algorithm (BP-ANN), so as to realize the qualitative identification of calamine samples. The accuracy rate of the model based on NIR and MRCC methods was 85%; in addition, the model, which took comprehensive multiple factors into consideration, can be used to identify crude calamine products, its counterfeits and processed products. Furthermore, by in-putting the correlation coefficients of multiple references as the spectral feature data of samples into BP-ANN, a BP-ANN model of qualitative identification was established, of which the accuracy rate was increased to 95%. The MRCC method can be used as a NIR-based method in the process of BP-ANN modeling.

  18. Hydrogen bonding in protic ionic liquids: structural correlations, vibrational spectroscopy, and rotational dynamics of liquid ethylammonium nitrate

    Zentel, Tobias; Overbeck, Viviane; Michalik, Dirk; Kühn, Oliver; Ludwig, Ralf

    2018-02-01

    The properties of the hydrogen bonds in ethylammonium nitrate (EAN) are analyzed by using molecular dynamics simulations and infrared as well as nuclear magnetic resonance experiments. EAN features a flexible three-dimensional network of hydrogen bonds with moderate strengths, which makes it distinct from related triethylammonium-based ionic liquids. First, the network’s flexibility is manifested in a not very pronounced correlation of the hydrogen bond geometries, which is caused by rapid interchanges of bonding partners. The large flexibility of the network also leads to a substantial broadening of the mid-IR absorption band, with the contributions due to N-H stretching motions ranging from 2800 to 3250 cm-1. Finally, the different dynamics are also seen in the rotational correlation of the N-H bond vector, where a correlation time as short as 16.1 ps is observed.

  19. Simultaneous 31P-NMR spectroscopy and EMG in exercising and recovering human skeletal muscle: a correlation study

    Vestergaard-Poulsen, P; Thomsen, C; Sinkjaer, T

    1995-01-01

    A large number of studies have shown amplitude and spectral changes of the electromyogram during exercise, leading to several theories of how these changes might be related to the underlying metabolic changes. The amplitude and spectral changes are generally interpreted as changes in motor unit...... of the muscle. Simultaneous 31P-nuclear magnetic resonance spectroscopy and surface electromyography were performed during sustained static exercise and recovery in healthy volunteers and a patient with McArdle's disease. A clear dissociation between the median power frequency of the surface electromyogram...... and pH was seen in the healthy volunteers during recovery and during exercise in the patient with McArdle's disease. The results indicate that proton or lactate accumulation is not primarily responsible for the spectral changes of the surface electromyogram as previously suggested. The motor unit...

  20. Accessibility of nucleic acid-complexed biomolecules to hydroxyl radicals correlates with their conformation: a fluorescence polarization spectroscopy study

    Makrigiorgos, G.M.; Bump, E.; Huang, C.; Kassis, A.I.; Baranowska-Kortylewicz, J.

    1994-01-01

    A fluorescence methodology has been developed to examine the relationship between the conformational state of specific biomolecules in simple chromatin models and their accessibility to hydroxyl radicals ( . OH). Polylysine and histone H1 were labelled with SECCA, the succinimidyl ester of coumarin-3-carboxylic acid, which generates the fluorescent derivative 7-OH-SECCA following its interaction with radiation-induced . OH in aqueous solution. The fluorescence induced per unit γ-ray dose reflecting the accessibility of . OH to such SECCA-conjugated biomolecules was recorded. The biomolecules were also labelled with the fluorescent derivative 7-OH-SECCA in trace amounts to study their conformation under identical conditions via fluorescence polarization spectroscopy. (author)