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Sample records for fluorescence based molecular

  1. Fluorescence based molecular in vivo imaging

    International Nuclear Information System (INIS)

    Ebert, Bernd

    2008-01-01

    Molecular imaging represents a modern research area that allows the in vivo study of molecular biological process kinetics using appropriate probes and visualization methods. This methodology may be defined- apart from the contrast media injection - as non-abrasive. In order to reach an in vivo molecular process imaging as accurate as possible the effects of the used probes on the biological should not be too large. The contrast media as important part of the molecular imaging can significantly contribute to the understanding of molecular processes and to the development of tailored diagnostics and therapy. Since more than 15 years PTB is developing optic imaging systems that may be used for fluorescence based visualization of tissue phantoms, small animal models and the localization of tumors and their predecessors, and for the early recognition of inflammatory processes in clinical trials. Cellular changes occur during many diseases, thus the molecular imaging might be of importance for the early diagnosis of chronic inflammatory diseases. Fluorescent dyes can be used as unspecific or also as specific contrast media, which allow enhanced detection sensitivity

  2. Highly Selective Fluorescent Sensing of Proteins Based on a Fluorescent Molecularly Imprinted Nanosensor

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    2013-09-01

    Full Text Available A fluorescent molecularly imprinted nanosensor was obtained by grafting imprinted polymer onto the surface of multi-wall carbon nanotubes and post-imprinting treatment with fluorescein isothiocyanate (FITC. The fluorescence of lysozyme-imprinted polymer (Lys-MIP was quenched more strongly by Lys than that of nonimprinted polymer (NIP, which indicated that the Lys-MIP could recognize Lys. The resulted imprinted material has the ability to selectively sense a target protein, and an imprinting factor of 3.34 was achieved. The Lys-MIP also showed selective detection for Lys among other proteins such as cytochrome C (Cyt C, hemoglobin (HB and bovine serum albumin (BSA due to the imprinted sites in the Lys-MIP. This approach combines the high selectivity of surface molecular imprinting technology and fluorescence, and converts binding events into detectable signals by monitoring fluorescence spectra. Therefore, it will have further applications for Lys sensing.

  3. Molecularly imprinted fluorescent probe based on FRET for selective and sensitive detection of doxorubicin

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Zhifeng, E-mail: 897061147@qq.com [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Deng, Peihong; Li, Junhua [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Xu, Li [Department of Applied Chemistry, College of Materials and Energy, South China Agricultural University, Guangzhou 510642 (China); Tang, Siping [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China)

    2017-04-15

    Highlights: • FRET-based molecularly imprinted probe for detection of doxorubicin was prepared. • The detection limit of the probe was 13.8 nM for doxorubicin. • The FRET-based probe had a higher selectivity for the template than ordinary MIMs. - Abstract: In this work, a new type of fluorescent probe for detection of doxorubicin has been constructed by the combined use of fluorescence resonance energy transfer (FRET) technology and molecular imprinting technique (MIT). Using doxorubicin as the template, the molecularly imprinted polymer thin layer was fabricated on the surfaces of carbon dot (CD) modified silica by sol-gel polymerization. The excitation energy of the fluorescent donor (CDs) could be transferred to the fluorescent acceptor (doxorubicin). The FRET based fluorescent probe demonstrated high sensitivity and selectivity for doxorubicin. The detection limit was 13.8 nM. The fluorescent probe was successfully applied for detecting doxorubicin in doxorubicin-spiked plasmas with a recovery of 96.8–103.8%, a relative standard deviation (RSD) of 1.3–2.8%. The strategy for construction of FRET-based molecularly imprinted materials developed in this work is very promising for analytical applications.

  4. Facilitating in vivo tumor localization by principal component analysis based on dynamic fluorescence molecular imaging

    Science.gov (United States)

    Gao, Yang; Chen, Maomao; Wu, Junyu; Zhou, Yuan; Cai, Chuangjian; Wang, Daliang; Luo, Jianwen

    2017-09-01

    Fluorescence molecular imaging has been used to target tumors in mice with xenograft tumors. However, tumor imaging is largely distorted by the aggregation of fluorescent probes in the liver. A principal component analysis (PCA)-based strategy was applied on the in vivo dynamic fluorescence imaging results of three mice with xenograft tumors to facilitate tumor imaging, with the help of a tumor-specific fluorescent probe. Tumor-relevant features were extracted from the original images by PCA and represented by the principal component (PC) maps. The second principal component (PC2) map represented the tumor-related features, and the first principal component (PC1) map retained the original pharmacokinetic profiles, especially of the liver. The distribution patterns of the PC2 map of the tumor-bearing mice were in good agreement with the actual tumor location. The tumor-to-liver ratio and contrast-to-noise ratio were significantly higher on the PC2 map than on the original images, thus distinguishing the tumor from its nearby fluorescence noise of liver. The results suggest that the PC2 map could serve as a bioimaging marker to facilitate in vivo tumor localization, and dynamic fluorescence molecular imaging with PCA could be a valuable tool for future studies of in vivo tumor metabolism and progression.

  5. Innovative molecular-based fluorescent nanoparticles for multicolor single particle tracking in cells

    International Nuclear Information System (INIS)

    Daniel, Jonathan; Blanchard-Desce, Mireille; Godin, Antoine G; Palayret, Matthieu; Lounis, Brahim; Cognet, Laurent

    2016-01-01

    Based on an original molecular-based design, we present bright and photostable fluorescent organic nanoparticles (FONs) showing excellent colloidal stability in various aqueous environments. Complementary near-infrared emitting and green emitting FONs were prepared using a simple, fast and robust protocol. Both types of FONs could be simultaneously imaged at the single-particle level in solution as well as in biological environments using a monochromatic excitation and a dual-color fluorescence microscope. No evidence of acute cytotoxicity was found upon incubation of live cells with mixed solutions of FONs, and both types of nanoparticles were found internalized in the cells where their motion could be simultaneously tracked at video-rate up to minutes. These fluorescent organic nanoparticles open a novel non-toxic alternative to existing nanoparticles for imaging biological structures, compatible with live-cell experiments and specially fitted for multicolor single particle tracking. (paper)

  6. Noncontact full-angle fluorescence molecular tomography system based on rotary mirrors.

    Science.gov (United States)

    Wang, Daifa; He, Jin; Qiao, Huiting; Li, Ping; Fan, Yubo; Li, Deyu

    2015-08-10

    We propose a novel noncontact fluorescence molecular tomography system that achieves full-angle capacity with the use of a new rotary-mirrors-based imaging head. In the imaging head, four plane mirrors are mounted on a rotating gantry to enable illumination and detection over 360°. In comparison with existing full-angle systems, our system does not require rotation of the specimen animal, a large and heavy light source (with scanning head), or a bulky camera (with filters and lens). The system design and implementation are described in detail. Both physical phantom and in vivo experiments are performed to verify the performance of the proposed system.

  7. Fluorescence- and capillary electrophoresis (CE)-based SSR DNA fingerprinting and a molecular identity database for the Louisiana sugarcane industry

    Science.gov (United States)

    A database of Louisiana sugarcane molecular identity has been constructed and is being updated annually using FAM or HEX or NED fluorescence- and capillary electrophoresis (CE)-based microsatellite (SSR) fingerprinting information. The fingerprints are PCR-amplified from leaf DNA samples of current ...

  8. MRI-guided fiber-based fluorescence molecular tomography for preclinical atherosclerosis imaging

    Science.gov (United States)

    Li, Baoqiang; Pouliot, Philippe; Lesage, Frederic

    2014-09-01

    Multi-modal imaging combining fluorescent molecular tomography (FMT) with MRI could provide information in these two modalities as well as optimize the recovery of functional information with MR-guidance. Here, we present a MRI-guided FMT system. An optical probe was designed consisting of a fiber plate on the top and bottom sides of the animal bed, respectively. In experiment, animal was installed between the two plates. Mounting fibers on each plate, transmission measuring could be conducted from both sides of the animal. Moreover, an accurate fluorescence reconstruction was achieved with MRI-derived anatomical guidance. The sensitivity of the FMT system was evaluated with a phantom showing that with long fibers, it was sufficient to detect 10nM Cy5.5 solution with ~28.5 dB in the phantom. The system was eventually used to image MMP activity involved in atherosclerosis with two ATX mice and two control mice. The reconstruction results were in agreement with ex vivo measurement.

  9. LED-FISH: Fluorescence microscopy based on light emitting diodes for the molecular analysis of Her-2/neu oncogene amplification

    Directory of Open Access Journals (Sweden)

    Vollmer Ekkehard

    2008-12-01

    Full Text Available Abstract Light emitting diodes (LED, which are available as small monochromatic light sources with characteristic features such as maximum illumination power combined with minimum energy consumption and extremely long lifespan have already proved as a highly potential low-cost alternative for specific diagnostic applications in clinical medicine such as tuberculosis fluorescence microscopy. Likewise, the most reliable evaluation of Her-2/neu (c-erbB2 gene amplification, which has been established in the last few years for routine diagnosis in clinical pathology as determinant towards Herceptin-based treatment of patients with breast cancer, is based on fluorescence in situ hybridization (FISH and corresponding high priced fluorescence equipment. In order to test the possibility to utilize the advantages of low-cost LED technology on FISH analysis of c-erbB2 gene expression for routine diagnostic purposes, the applicability of a standard bright field Carl Zeiss Axiostar Plus microscope equipped with a Fraen AFTER* LED Fluorescence Microscope Kit for the detection of Her-2/neu gene signals was compared to an advanced Nikon Eclipse 80i fluorescence microscope in combination with a conventional 100W mercury vapor lamp. Both microscopes were fitted with the same Quicam FAST CCD digital camera to unequivocally compare the quality of the captured images. C-erbB2 gene expression was analyzed in 30 different human tissue samples of primary invasive breast cancer, following formalin fixation and subsequent paraffin-embedding. The Her2/neu gene signals (green were identifiable in the tumor cells in all cases and images of equal quality were captured under almost identical conditions by 480 nm (blue LED module equipped standard Axiostar microscope as compared to conventional fluorescence microscopy. In this first attempt, these monochromatic LED elements proved in principle to be suitable for the detection of Her-2/neu gene expression by FISH. Thus, our own

  10. Molecular characteristics of a fluorescent chemosensor for the recognition of ferric ion based on photoresponsive azobenzene derivative

    Science.gov (United States)

    Chi, Zhen; Ran, Xia; Shi, Lili; Lou, Jie; Kuang, Yanmin; Guo, Lijun

    2017-01-01

    Metal ion recognition is of great significance in biological and environmental detection. So far, there is very few research related to the ferric ion sensing based on photoresponsive azobenzene derivatives. In this work, we report a highly selective fluorescent "turn-off" sensor for Fe3 + ions and the molecular sensing characteristics based on an azobenzene derivative, N-(3,4,5-octanoxyphenyl)-N‧-4-[(4-hydroxyphenyl)azophenyl]1,3,4-oxadiazole (AOB-t8). The binding association constant was determined to be 6.07 × 103 M- 1 in ethanol and the stoichiometry ratio of 2:2 was obtained from Job's plot and MS spectra. The AOB-t8 might be likely to form the dimer structure through the chelation of ferric ion with the azobenzene moiety. Meanwhile, it was found that the photoisomerization property of AOB-t8 was regulated by the binding with Fe3 +. With the chelation of Fe3 +, the regulated molecular rigidity and the perturbed of electronic state and molecular geometry was suggested to be responsible for the accelerated isomerization of AOB-t8 to UV irradiation and the increased fluorescence lifetime of both trans- and cis-AOB-t8-Fe(III). Moreover, the reversible sensing of AOB-t8 was successfully observed by releasing the iron ion from AOB-t8-Fe(III) with the addition of citric acid.

  11. Design and synthesis of a fluorescent molecular imprinted polymer for use in an optical fibre-based cocaine sensor

    Science.gov (United States)

    Wren, Stephen P.; Piletsky, Sergey A.; Karim, Kal; Gascoine, Paul; Lacey, Richard; Sun, Tong; Grattan, Kenneth T. V.

    2014-05-01

    Previously, we have developed chemical sensors using fibre optic-based techniques for the detection of Cocaine, utilising molecularly imprinted polymers (MIPs) containing fluorescein moieties as the signalling groups. Here, we report the computational design of a fluorophore which was incorporated into a MIP for the generation of a novel sensor that offers improved sensitivity for Cocaine with a detection range of 1-100μM. High selectivity for Cocaine over a suite of known Cocaine interferants (25μM) was also demonstrated by measuring changes in the intensity of fluorescence signals received from the sensor.

  12. Image reconstruction of fluorescent molecular tomography based on the tree structured Schur complement decomposition

    Directory of Open Access Journals (Sweden)

    Wang Jiajun

    2010-05-01

    Full Text Available Abstract Background The inverse problem of fluorescent molecular tomography (FMT often involves complex large-scale matrix operations, which may lead to unacceptable computational errors and complexity. In this research, a tree structured Schur complement decomposition strategy is proposed to accelerate the reconstruction process and reduce the computational complexity. Additionally, an adaptive regularization scheme is developed to improve the ill-posedness of the inverse problem. Methods The global system is decomposed level by level with the Schur complement system along two paths in the tree structure. The resultant subsystems are solved in combination with the biconjugate gradient method. The mesh for the inverse problem is generated incorporating the prior information. During the reconstruction, the regularization parameters are adaptive not only to the spatial variations but also to the variations of the objective function to tackle the ill-posed nature of the inverse problem. Results Simulation results demonstrate that the strategy of the tree structured Schur complement decomposition obviously outperforms the previous methods, such as the conventional Conjugate-Gradient (CG and the Schur CG methods, in both reconstruction accuracy and speed. As compared with the Tikhonov regularization method, the adaptive regularization scheme can significantly improve ill-posedness of the inverse problem. Conclusions The methods proposed in this paper can significantly improve the reconstructed image quality of FMT and accelerate the reconstruction process.

  13. Molecularly imprinted polymer based on MWCNT-QDs as fluorescent biomimetic sensor for specific recognition of target protein

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Zhaoqiang [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Annie Bligh, S.W. [Department of Life Sciences, Faculty of Science and Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW (United Kingdom); Tao, Lei; Quan, Jing [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Nie, Huali, E-mail: niehuali@dhu.edu.cn [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Zhu, Limin, E-mail: lzhu@dhu.edu.cn [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China); Gong, Xiao [College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620 (China)

    2015-03-01

    A novel molecularly imprinted optosensing material based on multi-walled carbon nanotube-quantum dots (MWCNT-QDs) has been designed and synthesized for its high selectivity, sensitivity and specificity in the recognition of a target protein bovine serum albumin (BSA). Molecularly imprinted polymer coated MWCNT-QDs using BSA as the template (BMIP-coated MWCNT-QDs) exhibits a fast mass-transfer speed with a response time of 25 min. It is found that the BSA as a target protein can significantly quench the luminescence of BMIP-coated MWCNT-QDs in a concentration-dependent manner that is best described by a Stern–Volmer equation. The K{sub SV} for BSA is much higher than bovine hemoglobin and lysozyme, implying a highly selective recognition of the BMIP-coated MWCNT-QDs to BSA. Under optimal conditions, the relative fluorescence intensity of BMIP-coated MWCNT-QDs decreases linearly with the increasing target protein BSA in the concentration range of 5.0 × 10{sup −7}–35.0 × 10{sup −7} M with a detection limit of 80 nM. - Highlights: • A novel fluorescent biomimetic sensor based on MWCNT-QDs was designed. • The sensor exhibited a fast mass-transfer speed with a response time of 25 min. • The sensor possessed a highly selective recognition to BSA.

  14. Molecularly imprinted polymer based on MWCNT-QDs as fluorescent biomimetic sensor for specific recognition of target protein

    International Nuclear Information System (INIS)

    Ding, Zhaoqiang; Annie Bligh, S.W.; Tao, Lei; Quan, Jing; Nie, Huali; Zhu, Limin; Gong, Xiao

    2015-01-01

    A novel molecularly imprinted optosensing material based on multi-walled carbon nanotube-quantum dots (MWCNT-QDs) has been designed and synthesized for its high selectivity, sensitivity and specificity in the recognition of a target protein bovine serum albumin (BSA). Molecularly imprinted polymer coated MWCNT-QDs using BSA as the template (BMIP-coated MWCNT-QDs) exhibits a fast mass-transfer speed with a response time of 25 min. It is found that the BSA as a target protein can significantly quench the luminescence of BMIP-coated MWCNT-QDs in a concentration-dependent manner that is best described by a Stern–Volmer equation. The K SV for BSA is much higher than bovine hemoglobin and lysozyme, implying a highly selective recognition of the BMIP-coated MWCNT-QDs to BSA. Under optimal conditions, the relative fluorescence intensity of BMIP-coated MWCNT-QDs decreases linearly with the increasing target protein BSA in the concentration range of 5.0 × 10 −7 –35.0 × 10 −7 M with a detection limit of 80 nM. - Highlights: • A novel fluorescent biomimetic sensor based on MWCNT-QDs was designed. • The sensor exhibited a fast mass-transfer speed with a response time of 25 min. • The sensor possessed a highly selective recognition to BSA

  15. Synchrotron-based X-ray Fluorescence Microscopy in Conjunction with Nanoindentation to Study Molecular-Scale Interactions of Phenol–Formaldehyde in Wood Cell Walls

    Science.gov (United States)

    Joseph E. Jakes; Christopher G. Hunt; Daniel J. Yelle; Linda Lorenz; Kolby Hirth; Sophie-Charlotte Gleber; Stefan Vogt; Warren Grigsby; Charles R. Frihart

    2015-01-01

    Understanding and controlling molecular-scale interactions between adhesives and wood polymers are critical to accelerate the development of improved adhesives for advanced wood-based materials. The submicrometer resolution of synchrotron-based X-ray fluorescence microscopy (XFM) was found capable of mapping and quantifying infiltration of Br-labeled phenol−...

  16. A quencher-free molecular beacon design based on pyrene excimer fluorescence using pyrene-labeled UNA (unlocked nucleic acid)

    DEFF Research Database (Denmark)

    Karlsen, Kasper Kannegård; Okholm, Anders Hauge; Kjems, Jørgen

    2013-01-01

    A quencher-free molecular beacon capable of generating pyrene excimer fluorescence has been constructed using strategically positioned pyrene-UNA monomers. Hybridization of a fully complementary RNA target was accompanied by a pyrene excimer emission increase of more than 900%, and detection of R...... in living cells was demonstrated....

  17. Mapping molecular assemblies with fluorescence microscopy and object-based spatial statistics.

    Science.gov (United States)

    Lagache, Thibault; Grassart, Alexandre; Dallongeville, Stéphane; Faklaris, Orestis; Sauvonnet, Nathalie; Dufour, Alexandre; Danglot, Lydia; Olivo-Marin, Jean-Christophe

    2018-02-15

    Elucidating protein functions and molecular organisation requires to localise precisely single or aggregated molecules and analyse their spatial distributions. We develop a statistical method SODA (Statistical Object Distance Analysis) that uses either micro- or nanoscopy to significantly improve on standard co-localisation techniques. Our method considers cellular geometry and densities of molecules to provide statistical maps of isolated and associated (coupled) molecules. We use SODA with three-colour structured-illumination microscopy (SIM) images of hippocampal neurons, and statistically characterise spatial organisation of thousands of synapses. We show that presynaptic synapsin is arranged in asymmetric triangle with the 2 postsynaptic markers homer and PSD95, indicating a deeper localisation of homer. We then determine stoichiometry and distance between localisations of two synaptic vesicle proteins with 3D-STORM. These findings give insights into the protein organisation at the synapse, and prove the efficiency of SODA to quantitatively assess the geometry of molecular assemblies.

  18. 3D mouse shape reconstruction based on phase-shifting algorithm for fluorescence molecular tomography imaging system.

    Science.gov (United States)

    Zhao, Yue; Zhu, Dianwen; Baikejiang, Reheman; Li, Changqing

    2015-11-10

    This work introduces a fast, low-cost, robust method based on fringe pattern and phase shifting to obtain three-dimensional (3D) mouse surface geometry for fluorescence molecular tomography (FMT) imaging. We used two pico projector/webcam pairs to project and capture fringe patterns from different views. We first calibrated the pico projectors and the webcams to obtain their system parameters. Each pico projector/webcam pair had its own coordinate system. We used a cylindrical calibration bar to calculate the transformation matrix between these two coordinate systems. After that, the pico projectors projected nine fringe patterns with a phase-shifting step of 2π/9 onto the surface of a mouse-shaped phantom. The deformed fringe patterns were captured by the corresponding webcam respectively, and then were used to construct two phase maps, which were further converted to two 3D surfaces composed of scattered points. The two 3D point clouds were further merged into one with the transformation matrix. The surface extraction process took less than 30 seconds. Finally, we applied the Digiwarp method to warp a standard Digimouse into the measured surface. The proposed method can reconstruct the surface of a mouse-sized object with an accuracy of 0.5 mm, which we believe is sufficient to obtain a finite element mesh for FMT imaging. We performed an FMT experiment using a mouse-shaped phantom with one embedded fluorescence capillary target. With the warped finite element mesh, we successfully reconstructed the target, which validated our surface extraction approach.

  19. Bilayered near-infrared fluorescent nanoparticles based on low molecular weight PEI for tumor-targeted in vivo imaging

    Science.gov (United States)

    Liu, Hao; Li, Ke; Xu, Liang; Wu, Daocheng

    2014-12-01

    To improve the tumor fluorescent imaging results in vivo, bilayered nanoparticles encapsulating a lipophilic near-infrared (NIR) fluorescent dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotri-carbocyanine iodide (DiR) were prepared using low molecular weight stearic acid-grafted polyethyleneimine and hyaluronic acid (DiR-PgSHA nanoparticles), which were investigated as a novel NIR fluorescent nano-probe for in vivo tumor-targeted optical imaging. These nanoparticles were characterized by transmission electron microscopy (TEM), infrared (IR) spectra, UV-visual absorption, and fluorescent emission spectra. Their cytotoxicity in vitro and hepatotoxicity in vivo were tested by MTT assay and histological study, respectively. In vivo NIR fluorescence imaging of the DiR-PgSHA nanoparticles was performed using a Carestream imaging system. The DiR-PgSHA nanoparticles were sphere shaped with a diameter of approximately 50 nm according to the TEM images. The DiR-PgSHA nanoparticles had a low cytotoxicity in vitro according to the MTT assay and low hepatotoxicity in vivo as determined in histological studies. The fluorescent emission of DiR-PgSHA nanoparticles was stable in pH values of 5-9 in solution, with only slight blue-shifts of the emission maxima at the basic pH range. The DiR-PgSHA nanoparticles exhibited a substantial tumor-targeting ability in the optical imaging with the use of tumor-bearing mice. These results demonstrated that the DiR-PgSHA nanoparticle is an excellent biocompatible nano-probe for in vivo tumor-targeted NIR fluorescence imaging with a potential for clinical applications.

  20. Bilayered near-infrared fluorescent nanoparticles based on low molecular weight PEI for tumor-targeted in vivo imaging

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Hao; Li, Ke [Xi’an Jiaotong University, Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology (China); Xu, Liang [The University of Kansas, Department of Molecular Biosciences (United States); Wu, Daocheng, E-mail: wudaocheng@mail.xjtu.edu.cn [Xi’an Jiaotong University, Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology (China)

    2014-12-15

    To improve the tumor fluorescent imaging results in vivo, bilayered nanoparticles encapsulating a lipophilic near-infrared (NIR) fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotri-carbocyanine iodide (DiR) were prepared using low molecular weight stearic acid-grafted polyethyleneimine and hyaluronic acid (DiR-PgSHA nanoparticles), which were investigated as a novel NIR fluorescent nano-probe for in vivo tumor-targeted optical imaging. These nanoparticles were characterized by transmission electron microscopy (TEM), infrared (IR) spectra, UV-visual absorption, and fluorescent emission spectra. Their cytotoxicity in vitro and hepatotoxicity in vivo were tested by MTT assay and histological study, respectively. In vivo NIR fluorescence imaging of the DiR-PgSHA nanoparticles was performed using a Carestream imaging system. The DiR-PgSHA nanoparticles were sphere shaped with a diameter of approximately 50 nm according to the TEM images. The DiR-PgSHA nanoparticles had a low cytotoxicity in vitro according to the MTT assay and low hepatotoxicity in vivo as determined in histological studies. The fluorescent emission of DiR-PgSHA nanoparticles was stable in pH values of 5–9 in solution, with only slight blue-shifts of the emission maxima at the basic pH range. The DiR-PgSHA nanoparticles exhibited a substantial tumor-targeting ability in the optical imaging with the use of tumor-bearing mice. These results demonstrated that the DiR-PgSHA nanoparticle is an excellent biocompatible nano-probe for in vivo tumor-targeted NIR fluorescence imaging with a potential for clinical applications.

  1. Reconstruction algorithms based on l1-norm and l2-norm for two imaging models of fluorescence molecular tomography: a comparative study.

    Science.gov (United States)

    Yi, Huangjian; Chen, Duofang; Li, Wei; Zhu, Shouping; Wang, Xiaorui; Liang, Jimin; Tian, Jie

    2013-05-01

    Fluorescence molecular tomography (FMT) is an important imaging technique of optical imaging. The major challenge of the reconstruction method for FMT is the ill-posed and underdetermined nature of the inverse problem. In past years, various regularization methods have been employed for fluorescence target reconstruction. A comparative study between the reconstruction algorithms based on l1-norm and l2-norm for two imaging models of FMT is presented. The first imaging model is adopted by most researchers, where the fluorescent target is of small size to mimic small tissue with fluorescent substance, as demonstrated by the early detection of a tumor. The second model is the reconstruction of distribution of the fluorescent substance in organs, which is essential to drug pharmacokinetics. Apart from numerical experiments, in vivo experiments were conducted on a dual-modality FMT/micro-computed tomography imaging system. The experimental results indicated that l1-norm regularization is more suitable for reconstructing the small fluorescent target, while l2-norm regularization performs better for the reconstruction of the distribution of fluorescent substance.

  2. A curcumin-based molecular probe for near-infrared fluorescence imaging of tau fibrils in Alzheimer's disease.

    Science.gov (United States)

    Park, Kwang-Su; Seo, Yujin; Kim, Mi Kyoung; Kim, Kyungdo; Kim, Yun Kyung; Choo, Hyunah; Chong, Youhoon

    2015-12-14

    In recent years, there has been growing interest in the near-infrared (NIR) fluorescence imaging of tau fibrils for the early diagnosis of Alzheimer's disease (AD). In order to develop a curcumin-based NIR fluorescent probe for tau fibrils, structural modification of the curcumin scaffold was attempted by combining the following rationales: the curcumin derivative should preserve its binding affinity to tau fibrils, and, upon binding to tau fibrils, the probe should show favorable fluorescence properties. To meet these requirements, we designed a novel curcumin scaffold with various aromatic substituents. Among the series, the curcumin derivative with a (4-dimethylamino-2,6-dimethoxy)phenyl moiety showed a significant change in its fluorescence properties (22.9-fold increase in quantum yield; Kd, 0.77 μM; λem, 620 nm; Φ, 0.32) after binding to tau fibrils. In addition, fluorescence imaging of tau-green fluorescent protein-transfected SHSY-5Y cells with confirmed that detected tau fibrils in live cells.

  3. A simple and sensitive surface molecularly imprinted polymers based fluorescence sensor for detection of λ-Cyhalothrin.

    Science.gov (United States)

    Liu, Chunbo; Song, Zhilong; Pan, Jianming; Yan, Yongsheng; Cao, Zhijing; Wei, Xiao; Gao, Lin; Wang, Juan; Dai, Jiangdong; Meng, Minjia; Yu, Ping

    2014-07-01

    In this study, surface molecularly imprinted YVO4:Eu(3+) nanoparticles with molecular recognitive optosensing activity were successfully prepared by precipitation polymerization using λ-Cyhalothrin (LC) as template molecules, methacrylic acid and ethylene glycol dimethacrylate as the polymerization precursors which could complex with template molecules, and the material has been characterized by SEM, TEM, FT-IR, XRD, TGA and so on. Meanwhile, the as-prepared core-shell structured nanocomposite (YVO4:Eu(3+)@MIPs), which was composed of lanthanide doped YVO4:Eu(3+) as fluorescent signal and surface molecular imprinted polymers as molecular selective recognition sites, could selectively and sensitively optosense the template molecules. After the experimental conditions were optimized, two linear relationship were obtained covering the concentration range of 2.0-10.0 μM and 10.0-90.0 μM, and the limit of detection (LOD) for LC was found to be 1.76 μM. Furthermore, a possible mechanism was put forward to explain the fluorescence quenching of YVO4:Eu(3+)@MIPs. More importantly, the obtained sensor was proven to be suitable for the detection of residues of LC in real examples. And the excellent performance of this sensor will facilitate future development of rapid and high-efficiency detection of LC. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Fluorescence lifetime based bioassays

    Science.gov (United States)

    Meyer-Almes, Franz-Josef

    2017-12-01

    Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

  5. Effective and robust approach for fluorescence molecular tomography based on CoSaMP and SP3 model

    Directory of Open Access Journals (Sweden)

    Xiaowei He

    2016-11-01

    Full Text Available Fluorescence molecular tomography (FMT allows the detection and quantification of various biological processes in small animals in vivo, which expands the horizons of pre-clinical research and drug development. Efficient three-dimensional (3D reconstruction algorithm is the key to accurate localization and quantification of fluorescent target in FMT. In this paper, 3D reconstruction of FMT is regarded as a sparse signal recovery problem and the compressive sampling matching pursuit (CoSaMP algorithm is adopted to obtain greedy recovery of fluorescent signals. Moreover, to reduce the modeling error, the simplified spherical harmonics approximation to the radiative transfer equation (RTE, more specifically SP3, is utilized to describe light propagation in biological tissues. The performance of the proposed reconstruction method is thoroughly evaluated by simulations on a 3D digital mouse model by comparing it with three representative greedy methods including orthogonal matching pursuit (OMP, stagewise OMP(StOMP, and regularized OMP (ROMP. The CoSaMP combined with SP3 shows an improvement in reconstruction accuracy and exhibits distinct advantages over the comparative algorithms in multiple targets resolving. Stability analysis suggests that CoSaMP is robust to noise and performs stably with reduction of measurements. The feasibility and reconstruction accuracy of the proposed method are further validated by phantom experimental data.

  6. Comprehensive phantom for interventional fluorescence molecular imaging.

    Science.gov (United States)

    Anastasopoulou, Maria; Koch, Maximilian; Gorpas, Dimitris; Karlas, Angelos; Klemm, Uwe; Garcia-Allende, Pilar Beatriz; Ntziachristos, Vasilis

    2016-09-01

    Fluorescence imaging has been considered for over a half-century as a modality that could assist surgical guidance and visualization. The administration of fluorescent molecules with sensitivity to disease biomarkers and their imaging using a fluorescence camera can outline pathophysiological parameters of tissue invisible to the human eye during operation. The advent of fluorescent agents that target specific cellular responses and molecular pathways of disease has facilitated the intraoperative identification of cancer with improved sensitivity and specificity over nonspecific fluorescent dyes that only outline the vascular system and enhanced permeability effects. With these new abilities come unique requirements for developing phantoms to calibrate imaging systems and algorithms. We briefly review herein progress with fluorescence phantoms employed to validate fluorescence imaging systems and results. We identify current limitations and discuss the level of phantom complexity that may be required for developing a universal strategy for fluorescence imaging calibration. Finally, we present a phantom design that could be used as a tool for interlaboratory system performance evaluation.

  7. Laser induced fluorescence of trapped molecular ions

    Energy Technology Data Exchange (ETDEWEB)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references.

  8. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references

  9. A molecular imprinting-based turn-on Ratiometric fluorescence sensor for highly selective and sensitive detection of 2,4-dichlorophenoxyacetic acid (2,4-D).

    Science.gov (United States)

    Wang, Xiaoyan; Yu, Jialuo; Wu, Xiaqing; Fu, Junqing; Kang, Qi; Shen, Dazhong; Li, Jinhua; Chen, Lingxin

    2016-07-15

    A novel molecular imprinting-based turn-on ratiometric fluorescence sensor was constructed via a facile sol-gel polymerization for detection of 2,4-dichlorophenoxyacetic acid (2,4-D) on the basis of photoinduced electron transfer (PET) by using nitrobenzoxadiazole (NBD) as detection signal source and quantum dots (QDs) as reference signal source. With the presence and increase of 2,4-D, the amine groups on the surface of QDs@SiO2 could bind with 2,4-D and thereby the NBD fluorescence intensities could be significantly enhanced since the PET process was inhibited, while the QDs maintained constant intensities. Accordingly, the ratio of the dual-emission intensities of green NBD and red QDs could be utilized for turn-on fluorescent detection of 2,4-D, along with continuous color changes from orange-red to green readily observed by the naked eye. The as-prepared fluorescence sensor obtained high sensitivity with a low detection limit of 0.14μM within 5min, and distinguished recognition selectivity for 2,4-D over its analogs. Moreover, the sensor was successfully applied to determine 2,4-D in real water samples, and high recoveries at three spiking levels of 2,4-D ranged from 95.0% to 110.1% with precisions below 4.5%. The simple, rapid and reliable visual sensing strategy would not only provide potential applications for high selective ultratrace analysis of complicated matrices, but also greatly enrich the research connotations of molecularly imprinted sensors. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Protein recognition by a pattern-generating fluorescent molecular probe

    Science.gov (United States)

    Pode, Zohar; Peri-Naor, Ronny; Georgeson, Joseph M.; Ilani, Tal; Kiss, Vladimir; Unger, Tamar; Markus, Barak; Barr, Haim M.; Motiei, Leila; Margulies, David

    2017-12-01

    Fluorescent molecular probes have become valuable tools in protein research; however, the current methods for using these probes are less suitable for analysing specific populations of proteins in their native environment. In this study, we address this gap by developing a unimolecular fluorescent probe that combines the properties of small-molecule-based probes and cross-reactive sensor arrays (the so-called chemical 'noses/tongues'). On the one hand, the probe can detect different proteins by generating unique identification (ID) patterns, akin to cross-reactive arrays. On the other hand, its unimolecular scaffold and selective binding enable this ID-generating probe to identify combinations of specific protein families within complex mixtures and to discriminate among isoforms in living cells, where macroscopic arrays cannot access. The ability to recycle the molecular device and use it to track several binding interactions simultaneously further demonstrates how this approach could expand the fluorescent toolbox currently used to detect and image proteins.

  11. Enhancing molecular logic through modulation of temporal and spatial constraints with quantum dot-based systems that use fluorescent (Förster) resonance energy transfer

    Science.gov (United States)

    Claussen, Jonathan C.; Algar, W. Russ; Hildebrandt, Niko; Susumu, Kimihiro; Ancona, Mario G.; Medintz, Igor L.

    2013-10-01

    Luminescent semiconductor nanocrystals or quantum dots (QDs) contain favorable photonic properties (e.g., resistance to photobleaching, size-tunable PL, and large effective Stokes shifts) that make them well-suited for fluorescence (Förster) resonance energy transfer (FRET) based applications including monitoring proteolytic activity, elucidating the effects of nanoparticles-mediated drug delivery, and analyzing the spatial and temporal dynamics of cellular biochemical processes. Herein, we demonstrate how unique considerations of temporal and spatial constraints can be used in conjunction with QD-FRET systems to open up new avenues of scientific discovery in information processing and molecular logic circuitry. For example, by conjugating both long lifetime luminescent terbium(III) complexes (Tb) and fluorescent dyes (A647) to a single QD, we can create multiple FRET lanes that change temporally as the QD acts as both an acceptor and donor at distinct time intervals. Such temporal FRET modulation creates multi-step FRET cascades that produce a wealth of unique photoluminescence (PL) spectra that are well-suited for the construction of a photonic alphabet and photonic logic circuits. These research advances in bio-based molecular logic open the door to future applications including multiplexed biosensing and drug delivery for disease diagnostics and treatment.

  12. Molecular cytogenetics using fluorescence in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Gray, J.W.; Kuo, Wen-Lin; Lucas, J.; Pinkel, D.; Weier, H-U.; Yu, Loh-Chung.

    1990-12-07

    Fluorescence in situ hybridization (FISH) with chromosome-specific probes enables several new areas of cytogenetic investigation by allowing visual determination of the presence and normality of specific genetic sequences in single metaphase or interphase cells. in this approach, termed molecular cytogenetics, the genetic loci to be analyzed are made microscopically visible in single cells using in situ hybridization with nucleic acid probes specific to these loci. To accomplish this, the DNA in the target cells is made single stranded by thermal denaturation and incubated with single-stranded, chemically modified probe under conditions where the probe will anneal only with DNA sequences to which it has high DNA sequence homology. The bound probe is then made visible by treatment with a fluorescent reagent such as fluorescein that binds to the chemical modification carried by the probe. The DNA to which the probe does not bind is made visible by staining with a dye such as propidium iodide that fluoresces at a wavelength different from that of the reagent used for probe visualization. We show in this report that probes are now available that make this technique useful for biological dosimetry, prenatal diagnosis and cancer biology. 31 refs., 3 figs.

  13. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Winn, J.S.

    1980-10-01

    Laser induced fluoresence (LIF) spectra (laser excitation spectra) are conceptually among the most simple spectra to obtain. One need only confine a gaseous sample in a suitable container, direct a laser along one axis of the container, and monitor the sample's fluorescence at a right angle to the laser beam. As the laser wavelength is changed, the changes in fluorescence intensity map the absorption spectrum of the sample. (More precisely, only absorption to states which have a significant radiative decay component are monitored.) For ion spectroscopy, one could benefit in many ways by such an experiment. Most optical ion spectra have been observed by emission techniques, and, aside from the problems of spectral analysis, discharge emission methods often produce the spectra of many species, some of which may be unknown or uncertain. Implicit in the description of LIF given above is certainty as to the chemical identity of the carrier of the spectrum. This article describes a method by which the simplifying aspects of LIF can be extended to molecular ions

  14. An ultrasensitive and highly selective fluorescent and colorimetric chemosensor for citrate ions based on rhodamine B and its application as the first molecular security keypad lock based on phosphomolybdic acid and citrate inputs

    International Nuclear Information System (INIS)

    Tavallali, Hossein; Baezzat, Mohammad-Reza; Deilamy-Rad, Gohar; Parhami, Abolfath; Hasanli, Nahid

    2015-01-01

    Rhodamine B (Rh B ) has been developed as novel and efficient colorimetric and fluorometric chemosensor for citrate ions (Cit 3− ) in an absolutely aqueous media. The UV–vis absorption and fluorescent emission titrations experiments have been employed to study the sensing process. Rh B could act as an efficient “ON–OFF” fluorescent chemosensor for phosphomolybdic acid (PMA) based on an electron transfer (ET) process. Also (Rh B + ) 3 .PMA 3− could operate as an “OFF–ON” fluorescent chemosensor for citrate ions based on a ligand substitution process. The chemosensor Rh B shows excellent fluorescence sensitivity and selectivity toward citrate in aqueous media, and displays ON–OFF–ON type fluorescence change with alternately adding PMA and citrate to the media along with reversible association–dissociation of the complex. The (Rh B + ) 3 .PMA 3− can be applied to the quantification of citrate with a linear ranges covering from 0.053 to 0.83 and 0.08 to 1.6 µM by detection limits of 6.0 and 9.1 nM for fluorescence and colorimetric methods respectively. The keypad lock operation is particularly important, as the output of the system depends not only on the proper combination but also on the order of input signals, creating the correct password that can be used to “open” this molecular keypad lock through strong fluorescence emission at 575 nm. As a whole, its various logic gate properties may improve its impact for the development of new-generation “intelligence” digital devices. The ionic PMA and Cit 3− inputs to (Rh B + ) 3 .PMA 3− have been mimicked as a superimposed electronic molecular keypad lock. Also indicates that Rh B is suitable for the detection of Cit 3− ions in the biological environment. - Highlights: • Our probe is commercially available with good photostability and high quantum yield. • Both color and fluorescence change with long emission wavelength in aqueous media. • Characteristics of an ON

  15. Polarization of fluorescence: a probe of molecular autoionization

    International Nuclear Information System (INIS)

    Leroi, G.E.; Dehmer, J.L.; Parr, A.C.; Poliakoff, E.D.

    1983-01-01

    The polarization of fluorescence from excited-state molecular photoions provides a direct probe of the photoionization dynamics and the symmetry signatures of autoionizing resonances. Measurements on CO 2 and CS 2 are presented as examples

  16. Multiscale Humidity Visualization by Environmentally Sensitive Fluorescent Molecular Rotors.

    Science.gov (United States)

    Cheng, Yanhua; Wang, Jianguo; Qiu, Zijie; Zheng, Xiaoyan; Leung, Nelson L C; Lam, Jacky W Y; Tang, Ben Zhong

    2017-12-01

    Building humidity sensors possessing the features of diverse-configuration compatibility, and capability of measurement of spatial and temporal humidity gradients is of great interest for highly integrated electronics and wearable monitoring systems. Herein, a visual sensing approach based on fluorescent imaging is presented, by assembling aggregation-induced-emission (AIE)-active molecular rotors into a moisture-captured network; the resulting AIE humidity sensors are compatible with diverse applications, having tunable geometries and desirable architectures. The invisible information of relative humidity (RH) is transformed into different fluorescence colors that enable direct observation by the naked eyes based on the twisted intramolecular charge-transfer effect of the AIE-active molecular rotors. The resulting AIE humidity sensors show excellent performance in terms of good sensitivity, precise quantitative measurement, high spatial-temporal resolution, and fast response/recovery time. Their multiscale applications, such as regional environmental RH detection, internal humidity mapping, and sensitive human-body humidity sensing are demonstrated. The proposed humidity visualization strategy may provide a new insight to develop humidity sensors for various applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Chemical point detection using differential fluorescence from molecularly imprinted polymers

    Science.gov (United States)

    Pestov, Dmitry; Anderson, John E.; Nelson, Jean; Tepper, Gary C.

    2004-12-01

    Fluorescence represents one of the most attractive approaches for chemical sensing due to the abundant light produced by most fluorophores, resulting in excellent detection sensitivity. However, the broad and overlapping emission spectra of target and background species have made it difficult to perform species identification in a field instrument because of the need to perform spectral decomposition and analysis. This paper describes a new chemical sensing strategy based on differential fluorescence measurements from molecularly imprinted polymers, which eliminates the need to perform any spectral analysis. Species identification is accomplished by measuring the differential light output from a pair of polymers-one imprinted to a target species and the other identical, but not imprinted. The imprinted polymer selectively concentrates the target molecule and controls the energy (wavelength) of the emitted fluorescence signal and the differential output eliminates common mode signals associated with non-specific background interference. Because no spectral analysis is required, the sensors can be made extremely small and require very little power. Preliminary performance parameters from a prototype sensor are presented and discussed.

  18. Time-Resolved Fluorescence Anisotropy of Bicyclo[1.1.1]pentane/Tolane-Based Molecular Rods Included in Tris(o-phenylenedioxy)cyclotriphosphazene (TPP).

    Science.gov (United States)

    Cipolloni, Marco; Kaleta, Jiří; Mašát, Milan; Dron, Paul I; Shen, Yongqiang; Zhao, Ke; Rogers, Charles T; Shoemaker, Richard K; Michl, Josef

    2015-04-23

    We examine the fluorescence anisotropy of rod-shaped guests held inside the channels of tris( o -phenylenedioxy)cyclotriphosphazene (TPP) host nanocrystals, characterized by powder X-ray diffraction and solid state NMR spectroscopy. We address two issues: (i) are light polarization measurements on an aqueous colloidal solution of TPP nanocrystals meaningful, or is depolarization by scattering excessive? (ii) Can measurements of the rotational mobility of the included guests be performed at low enough loading levels to suppress depolarization by intercrystallite energy transfer? We find that meaningful measurements are possible and demonstrate that the long axis of molecular rods included in TPP channels performs negligible vibrational motion.

  19. Molecular Iodine Fluorescence Using a Green Helium-Neon Laser

    Science.gov (United States)

    Williamson, J. Charles

    2011-01-01

    Excitation of molecular iodine vapor with a green (543.4 nm) helium-neon laser produces a fluorescence spectrum that is well suited for the upper-level undergraduate physical chemistry laboratory. Application of standard evaluation techniques to the spectrum yields ground electronic-state molecular parameters in good agreement with literature…

  20. Laplacian manifold regularization method for fluorescence molecular tomography

    Science.gov (United States)

    He, Xuelei; Wang, Xiaodong; Yi, Huangjian; Chen, Yanrong; Zhang, Xu; Yu, Jingjing; He, Xiaowei

    2017-04-01

    Sparse regularization methods have been widely used in fluorescence molecular tomography (FMT) for stable three-dimensional reconstruction. Generally, ℓ1-regularization-based methods allow for utilizing the sparsity nature of the target distribution. However, in addition to sparsity, the spatial structure information should be exploited as well. A joint ℓ1 and Laplacian manifold regularization model is proposed to improve the reconstruction performance, and two algorithms (with and without Barzilai-Borwein strategy) are presented to solve the regularization model. Numerical studies and in vivo experiment demonstrate that the proposed Gradient projection-resolved Laplacian manifold regularization method for the joint model performed better than the comparative algorithm for ℓ1 minimization method in both spatial aggregation and location accuracy.

  1. Exploiting Molecular Biology by Time-Resolved Fluorescence Imaging

    Science.gov (United States)

    Müller, Francis; Fattinger, Christof

    Many contemporary biological investigations rely on highly sensitive in vitro assays for the analysis of specific molecules in biological specimens, and the main part of these assays depends on high-sensitivity fluorescence detection techniques for the final readout. The analyzed molecules and molecular interactions in the specimen need to be detected in the presence of other highly abundant biomolecules, while the analyzed molecules themselves are only present at nano-, pico-, or even femtomolar concentration.A short scientific rationale of fluorescence is presented. It emphasizes the use of fluorescent labels for sensitive assays in life sciences and specifies the main properties of an ideal fluorophore. With fluorescence lifetimes in the microsecond range and fluorescence quantum yield of 0.4 some water soluble complexes of Ruthenium like modified Ru(sulfobathophenanthroline) complexes fulfill these properties. They are outstanding fluorescent labels for ultrasensitive assays as illustrated in two examples, in drug discovery and in point of care testing.We discuss the fundamentals and the state-of-the-art of the most sensitive time-gated fluorescence assays. We reflect on how the imaging devices currently employed for readout of these assays might evolve in the future. Many contemporary biological investigations rely on highly sensitive in vitro assays for the analysis of specific molecules in biological specimens, and the main part of these assays depends on high-sensitivity fluorescence detection techniques for the final readout. The analyzed molecules and molecular interactions in the specimen need to be detected in the presence of other highly abundant biomolecules, while the analyzed molecules themselves are only present at nano-, pico-, or even femtomolar concentration.A short scientific rationale of fluorescence is presented. It emphasizes the use of fluorescent labels for sensitive assays in life sciences and specifies the main properties of an ideal

  2. Ratiometric Molecular Probes Based on Dual Emission of a Blue Fluorescent Coumarin and a Red Phosphorescent Cationic Iridium(III) Complex for Intracellular Oxygen Sensing

    Science.gov (United States)

    Yoshihara, Toshitada; Murayama, Saori; Tobita, Seiji

    2015-01-01

    Ratiometric molecular probes RP1 and RP2 consisting of a blue fluorescent coumarin and a red phosphorescent cationic iridium complex connected by a tetra- or octaproline linker, respectively, were designed and synthesized for sensing oxygen levels in living cells. These probes exhibited dual emission with good spectral separation in acetonitrile. The photorelaxation processes, including intramolecular energy transfer, were revealed by emission quantum yield and lifetime measurements. The ratios (RI=(Ip/If)) between the phosphorescence (Ip) and fluorescence (If) intensities showed excellent oxygen responses; the ratio of RI under degassed and aerated conditions (RI0/RI) was 20.3 and 19.6 for RP1 and RP2. The introduction of the cationic Ir (III) complex improved the cellular uptake efficiency compared to that of a neutral analogue with a tetraproline linker. The emission spectra of the ratiometric probes internalized into living HeLa or MCF-7 cells could be obtained using a conventional microplate reader. The complex RP2 with an octaproline linker provided ratios comparable to the ratiometric measurements obtained using a microplate reader: the ratio of the RI value of RP2 under hypoxia (2.5% O2) to that under normoxia (21% O2) was 1.5 and 1.7 for HeLa and MCF-7 cells, respectively. Thus, the intracellular oxygen levels of MCF-7 cells could be imaged by ratiometric emission measurements using the complex RP2. PMID:26066988

  3. Dual-tracer background subtraction approach for fluorescent molecular tomography

    Science.gov (United States)

    Holt, Robert W.; El-Ghussein, Fadi; Davis, Scott C.; Samkoe, Kimberley S.; Gunn, Jason R.; Leblond, Frederic

    2013-01-01

    Abstract. Diffuse fluorescence tomography requires high contrast-to-background ratios to accurately reconstruct inclusions of interest. This is a problem when imaging the uptake of fluorescently labeled molecularly targeted tracers in tissue, which can result in high levels of heterogeneously distributed background uptake. We present a dual-tracer background subtraction approach, wherein signal from the uptake of an untargeted tracer is subtracted from targeted tracer signal prior to image reconstruction, resulting in maps of targeted tracer binding. The approach is demonstrated in simulations, a phantom study, and in a mouse glioma imaging study, demonstrating substantial improvement over conventional and homogenous background subtraction image reconstruction approaches. PMID:23292612

  4. Environment-sensitive behavior of fluorescent molecular rotors

    Directory of Open Access Journals (Sweden)

    Theodorakis Emmanuel A

    2010-09-01

    Full Text Available Abstract Molecular rotors are a group of fluorescent molecules that form twisted intramolecular charge transfer (TICT states upon photoexcitation. When intramolecular twisting occurs, the molecular rotor returns to the ground state either by emission of a red-shifted emission band or by nonradiative relaxation. The emission properties are strongly solvent-dependent, and the solvent viscosity is the primary determinant of the fluorescent quantum yield from the planar (non-twisted conformation. This viscosity-sensitive behavior gives rise to applications in, for example, fluid mechanics, polymer chemistry, cell physiology, and the food sciences. However, the relationship between bulk viscosity and the molecular-scale interaction of a molecular rotor with its environment are not fully understood. This review presents the pertinent theories of the rotor-solvent interaction on the molecular level and how this interaction leads to the viscosity-sensitive behavior. Furthermore, current applications of molecular rotors as microviscosity sensors are reviewed, and engineering aspects are presented on how measurement accuracy and precision can be improved.

  5. Ratiometric Molecular Probes Based on Dual Emission of a Blue Fluorescent Coumarin and a Red Phosphorescent Cationic Iridium(III Complex for Intracellular Oxygen Sensing

    Directory of Open Access Journals (Sweden)

    Toshitada Yoshihara

    2015-06-01

    Full Text Available Ratiometric molecular probes RP1 and RP2 consisting of a blue fluorescent coumarin and a red phosphorescent cationic iridium complex connected by a tetra- or octaproline linker, respectively, were designed and synthesized for sensing oxygen levels in living cells. These probes exhibited dual emission with good spectral separation in acetonitrile. The photorelaxation processes, including intramolecular energy transfer, were revealed by emission quantum yield and lifetime measurements. The ratios (RI = (Ip /If between the phosphorescence (Ip and fluorescence (If intensities showed excellent oxygen responses; the ratio of RI under degassed and aerated conditions ( R I 0 MathType@MTEF@5@5@+= feaagKart1ev2aqatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr 4rNCHbGeaGqiVCI8FfYJH8YrFfeuY=Hhbbf9v8qqaqFr0xc9pk0xbb a9q8WqFfeaY=biLkVcLq=JHqpepeea0=as0Fb9pgeaYRXxe9vr0=vr 0=vqpWqaaeaabiGaciaacaqabeaadaqaaqaaaOqaaabaaaaaaaaape GaamOua8aadaqhaaWcbaWdbiaadMeaa8aabaWdbiaaicdaaaaaaa@38D6@ / RI was 20.3 and 19.6 for RP1 and RP2. The introduction of the cationic Ir (III complex improved the cellular uptake efficiency compared to that of a neutral analogue with a tetraproline linker. The emission spectra of the ratiometric probes internalized into living HeLa or MCF-7 cells could be obtained using a conventional microplate reader. The complex RP2 with an octaproline linker provided ratios comparable to the ratiometric measurements obtained using a microplate reader: the ratio of the  value of RP2 under hypoxia (2.5% O2 to that under normoxia (21% O2 was 1.5 and 1.7 for HeLa and MCF-7 cells, respectively. Thus, the intracellular oxygen levels of MCF-7 cells could be imaged by ratiometric emission measurements using the complex RP2.

  6. Molecular engineering and fluorescence for the detection of toxic cations

    International Nuclear Information System (INIS)

    Souchon, V.

    2007-11-01

    This work is a part of the 'Toxicologie Nucleaire Environnementale' program which aims at studying the effects on the living of heavy metals or radionuclides involved in nuclear industry. Most particularly, it deals with the design of new fluorescent sensors for the selective detection of Pb 2+ , Cd 2+ and Cs + in biological media. Several fluorescent calixarenes possessing nitrogen atoms were synthesized and their properties as potential lead sensors were investigated. One of them could be used in experimental conditions close to biological media and new target compounds with amide functional groups were proposed. Many approaches were considered for the design of selective fluorescent sensors for cadmium. On the basis of literature results, many chelating compounds incorporating sulfur atoms were synthesized but showed no significant affinity towards cadmium. On the opposite, compounds functionalized with several pyridine-2'-yl-1,2,3-triazol fluorescent moieties linked to a β-cyclodextrin or a calix[4]arene showed good affinity for cadmium in methanol, but the selectivity was found to be insufficient. In contrast, very satisfying results in terms of both selectivity and sensitivity could be obtained with the commercial calcium sensor Rhod-5N in an aqueous medium at neutral pH. Lastly, micromolar detection limits for the selective detection of caesium were reached in an aqueous medium at neutral pH thanks to a new sulfonated fluorescent calixarene with two appended crown-ethers. An original complexation mechanism was proposed and validated by molecular modelling (DFT). (author)

  7. In Vivo Diffuse Optical Tomography and Fluorescence Molecular Tomography

    Directory of Open Access Journals (Sweden)

    Mingze Li

    2010-01-01

    Full Text Available Diffuse optical tomography (DOT and fluorescence molecular tomography (FMT are two attractive imaging techniques for in vivo physiological and psychological research. They have distinct advantages such as non-invasiveness, non-ionizing radiation, high sensitivity and longitudinal monitoring. This paper reviews the key components of DOT and FMT. Light propagation model, mathematical reconstruction algorithm, imaging instrumentation and medical applications are included. Future challenges and perspective on optical tomography are discussed.

  8. Fluorescence excitation studies of molecular photoionization in external electric fields

    International Nuclear Information System (INIS)

    Poliakoff, E.D.; Dehmer, J.L.; Parr, A.C.; Leroi, G.E.

    1985-01-01

    Using molecular nitrogen as an example, we show that fluorescence excitation spectroscopy can be used to measure partial photoionization cross sections of free molecules in external electric fields. The production of the N 2 + (B 2 Σ/sub u/ + ) state was studied and the threshold for this process was found to shift linearly with the square root of the applied field. This behavior is compared with the hydrogenic case and with previously studied systems

  9. Comparison of Regularization Methods in Fluorescence Molecular Tomography

    Directory of Open Access Journals (Sweden)

    Dianwen Zhu

    2014-04-01

    Full Text Available In vivo fluorescence molecular tomography (FMT has been a popular functional imaging modality in research labs in the past two decades. One of the major difficulties of FMT lies in the ill-posed and ill-conditioned nature of the inverse problem in reconstructing the distribution of fluorophores inside objects. The popular regularization methods based on L2, L1 and total variation (TV norms have been applied in FMT reconstructions. The non-convex Lq(0 < q < 1 semi-norm and Log function have also been studied recently. In this paper, we adopt a uniform optimization transfer framework for these regularization methods in FMT and compare their individual, as well as the combined effects on both small, localized targets, such as tumors in the early stage, and large targets, such as liver. Numerical simulation studies and phantom experiments have been carried out, and we found that Lq with q near 1/2 performs the best in reconstructing small targets, while joint L2 and Log performs the best for large targets.

  10. Nucleic acid-based fluorescent probes and their analytical potential.

    Science.gov (United States)

    Juskowiak, Bernard

    2011-03-01

    It is well known that nucleic acids play an essential role in living organisms because they store and transmit genetic information and use that information to direct the synthesis of proteins. However, less is known about the ability of nucleic acids to bind specific ligands and the application of oligonucleotides as molecular probes or biosensors. Oligonucleotide probes are single-stranded nucleic acid fragments that can be tailored to have high specificity and affinity for different targets including nucleic acids, proteins, small molecules, and ions. One can divide oligonucleotide-based probes into two main categories: hybridization probes that are based on the formation of complementary base-pairs, and aptamer probes that exploit selective recognition of nonnucleic acid analytes and may be compared with immunosensors. Design and construction of hybridization and aptamer probes are similar. Typically, oligonucleotide (DNA, RNA) with predefined base sequence and length is modified by covalent attachment of reporter groups (one or more fluorophores in fluorescence-based probes). The fluorescent labels act as transducers that transform biorecognition (hybridization, ligand binding) into a fluorescence signal. Fluorescent labels have several advantages, for example high sensitivity and multiple transduction approaches (fluorescence quenching or enhancement, fluorescence anisotropy, fluorescence lifetime, fluorescence resonance energy transfer (FRET), and excimer-monomer light switching). These multiple signaling options combined with the design flexibility of the recognition element (DNA, RNA, PNA, LNA) and various labeling strategies contribute to development of numerous selective and sensitive bioassays. This review covers fundamentals of the design and engineering of oligonucleotide probes, describes typical construction approaches, and discusses examples of probes used both in hybridization studies and in aptamer-based assays.

  11. Improved sparse reconstruction for fluorescence molecular tomography with L1/2 regularization.

    Science.gov (United States)

    Guo, Hongbo; Yu, Jingjing; He, Xiaowei; Hou, Yuqing; Dong, Fang; Zhang, Shuling

    2015-05-01

    Fluorescence molecular tomography (FMT) is a promising imaging technique that allows in vivo visualization of molecular-level events associated with disease progression and treatment response. Accurate and efficient 3D reconstruction algorithms will facilitate the wide-use of FMT in preclinical research. Here, we utilize L1/2-norm regularization for improving FMT reconstruction. To efficiently solve the nonconvex L1/2-norm penalized problem, we transform it into a weighted L1-norm minimization problem and employ a homotopy-based iterative reweighting algorithm to recover small fluorescent targets. Both simulations on heterogeneous mouse model and in vivo experiments demonstrated that the proposed L1/2-norm method outperformed the comparative L1-norm reconstruction methods in terms of location accuracy, spatial resolution and quantitation of fluorescent yield. Furthermore, simulation analysis showed the robustness of the proposed method, under different levels of measurement noise and number of excitation sources.

  12. Mapping microbubble viscosity using fluorescence lifetime imaging of molecular rotors

    Science.gov (United States)

    Hosny, Neveen A.; Mohamedi, Graciela; Rademeyer, Paul; Owen, Joshua; Wu, Yilei; Tang, Meng-Xing; Eckersley, Robert J.; Stride, Eleanor; Kuimova, Marina K.

    2013-01-01

    Encapsulated microbubbles are well established as highly effective contrast agents for ultrasound imaging. There remain, however, some significant challenges to fully realize the potential of microbubbles in advanced applications such as perfusion mapping, targeted drug delivery, and gene therapy. A key requirement is accurate characterization of the viscoelastic surface properties of the microbubbles, but methods for independent, nondestructive quantification and mapping of these properties are currently lacking. We present here a strategy for performing these measurements that uses a small fluorophore termed a “molecular rotor” embedded in the microbubble surface, whose fluorescence lifetime is directly related to the viscosity of its surroundings. We apply fluorescence lifetime imaging to show that shell viscosities vary widely across the population of the microbubbles and are influenced by the shell composition and the manufacturing process. We also demonstrate that heterogeneous viscosity distributions exist within individual microbubble shells even with a single surfactant component. PMID:23690599

  13. Molecular evidence of Opisthorchis viverrini in infected bithyniid snails in the Lao People's Democratic Republic by specific hybridization probe-based real-time fluorescence resonance energy transfer PCR method.

    Science.gov (United States)

    Sri-Aroon, Pusadee; Intapan, Pewpan M; Lohachit, Chantima; Phongsasakulchoti, Phunthira; Thanchomnang, Tongjit; Lulitanond, Viraphong; Hiscox, Alexandra; Phompida, Samlane; Sananikhom, Pany; Maleewong, Wanchai; Brey, Paul T

    2011-04-01

    Naturally occurring bithyniid snails, Bithynia siamensis goniomphalos (Prosobranchia: Bithyniidae), and their intermediate hosts were sampled from Khammouane Province, the Lao People's Democratic Republic, and the prevalence of the carcinogenic human liver fluke, Opisthorchis viverrini, was examined. The presence of O. viverrini cercariae in snails was examined by cercarial shedding test and then confirmed by specific hybridization probe-based real-time fluorescence resonance energy transfer (FRET) PCR method. The real-time FRET PCR method is based on a fluorescence melting curve analysis of a hybrid between an amplicon produced from the pOV-A6 specific sequence (Genbank accession no. S80278), a 162-bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. Mean melting temperature of O. viverrini DNA from the cercariae and each of two positive snails by shedding test was 66.3 ± 0.1. The O. viverrini infection rate in snails was 2.47% (2/81) by cercarial shedding test but was 8.52% (4/47) by real-time FRET PCR method. The real-time FRET PCR method is rapid and effective in examining a large number of snail samples simultaneously. Validation using molecular evidence from this procedure provides another tool for surveying the prevalence of O. viverrini-infected snails in Southeast Asian countries.

  14. Counting constituents in molecular complexes by fluorescence photon antibunching

    Energy Technology Data Exchange (ETDEWEB)

    Fore, S; Laurence, T; Hollars, C; Huser, T

    2007-04-17

    Modern single molecule fluorescence microscopy offers new, highly quantitative ways of studying the systems biology of cells while keeping the cells healthy and alive in their natural environment. In this context, a quantum optical technique, photon antibunching, has found a small niche in the continuously growing applications of single molecule techniques to small molecular complexes. Here, we review some of the most recent applications of photon antibunching in biophotonics, and we provide a guide for how to conduct photon antibunching experiments at the single molecule level by applying techniques borrowed from time-correlated single photon counting. We provide a number of new examples for applications of photon antibunching to the study of multichromophoric molecules and small molecular complexes.

  15. Novel biosensor system model based on fluorescence quenching by a fluorescent streptavidin and carbazole-labeled biotin.

    Science.gov (United States)

    Zhu, Xianwei; Shinohara, Hiroaki; Miyatake, Ryuta; Hohsaka, Takahiro

    2016-10-01

    In the present study, a novel molecular biosensor system model was designed by using a couple of the fluorescent unnatural mutant streptavidin and the carbazole-labeled biotin. BODIPY-FL-aminophenylalanine (BFLAF), a fluorescent unnatural amino acid was position-specifically incorporated into Trp120 position of streptavidin by four-base codon method. On the other hand, carbazole-labeled biotin was synthesized as a quencher for the fluorescent Trp120BFLAF mutant streptavidin. The fluorescence of fluorescent Trp120BFLAF mutant streptavidin was decreased as we expected when carbazole-labeled biotin was added into the mutant streptavidin solution. Furthermore, the fluorescence decrease of Trp120BFLAF mutant streptavidin with carbazole-labeled biotin (100 nM) was recovered by the competitive addition of natural biotin. This result demonstrated that by measuring the fluorescence quenching and recovery, a couple of the fluorescent Trp120BFLAF mutant streptavidin and the carbazole-labeled biotin were successfully applicable for quantification of free biotin as a molecular biosensor system. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  16. Fluorescence quantum yield measurements of fluorescent proteins: a laboratory experiment for a biochemistry or molecular biophysics laboratory course.

    Science.gov (United States)

    Wall, Kathryn P; Dillon, Rebecca; Knowles, Michelle K

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts absorbed photons into emitted photons and it is necessary to know for assessing what fluorescent protein is the most appropriate for a particular application. In this work, we have designed an upper-level, biochemistry laboratory experiment where students measure the fluorescence quantum yields of fluorescent proteins relative to a standard organic dye. Four fluorescent protein variants, enhanced cyan fluorescent protein (ECFP), enhanced green fluorescent protein (EGFP), mCitrine, and mCherry, were used, however the methods described are useful for the characterization of any fluorescent protein or could be expanded to fluorescent quantum yield measurements of organic dye molecules. The laboratory is designed as a guided inquiry project and takes two, 4 hr laboratory periods. During the first day students design the experiment by selecting the excitation wavelength, choosing the standard, and determining the concentration needed for the quantum yield experiment that takes place in the second laboratory period. Overall, this laboratory provides students with a guided inquiry learning experience and introduces concepts of fluorescence biophysics into a biochemistry laboratory curriculum. © 2014 The International Union of Biochemistry and Molecular Biology.

  17. Nanoplatform-based molecular imaging

    National Research Council Canada - National Science Library

    Chen, Xiaoyuan

    2011-01-01

    "Nanoplathform-Based Molecular Imaging provides rationale for using nanoparticle-based probes for molecular imaging, then discusses general strategies for this underutilized, yet promising, technology...

  18. Molecular engineering of two-photon fluorescent probes for bioimaging applications

    Science.gov (United States)

    Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

    2017-03-01

    During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

  19. Excitation-resolved multispectral method for imaging pharmacokinetic parameters in dynamic fluorescent molecular tomography

    Science.gov (United States)

    Chen, Maomao; Zhou, Yuan; Su, Han; Zhang, Dong; Luo, Jianwen

    2017-04-01

    Imaging of the pharmacokinetic parameters in dynamic fluorescence molecular tomography (DFMT) can provide three-dimensional metabolic information for biological studies and drug development. However, owing to the ill-posed nature of the FMT inverse problem, the relatively low quality of the parametric images makes it difficult to investigate the different metabolic processes of the fluorescent targets with small distances. An excitation-resolved multispectral DFMT method is proposed; it is based on the fact that the fluorescent targets with different concentrations show different variations in the excitation spectral domain and can be considered independent signal sources. With an independent component analysis method, the spatial locations of different fluorescent targets can be decomposed, and the fluorescent yields of the targets at different time points can be recovered. Therefore, the metabolic process of each component can be independently investigated. Simulations and phantom experiments are carried out to evaluate the performance of the proposed method. The results demonstrated that the proposed excitation-resolved multispectral method can effectively improve the reconstruction accuracy of the parametric images in DFMT.

  20. Molecular quantification of genes encoding for green-fluorescent proteins

    DEFF Research Database (Denmark)

    Felske, A; Vandieken, V; Pauling, B V

    2003-01-01

    A quantitative PCR approach is presented to analyze the amount of recombinant green fluorescent protein (gfp) genes in environmental DNA samples. The quantification assay is a combination of specific PCR amplification and temperature gradient gel electrophoresis (TGGE). Gene quantification...... is provided by a competitively coamplified DNA standard constructed by point mutation PCR. A single base difference was introduced to achieve a suitable migration difference in TGGE between the original target DNA and the modified standard without altering the PCR amplification efficiency. This competitive...... PCR strategy is a highly specific and sensitive way to monitor recombinant DNA in environments like the efflux of a biotechnological plant....

  1. Fluorescent Biosensors Based on Single-Molecule Counting.

    Science.gov (United States)

    Ma, Fei; Li, Ying; Tang, Bo; Zhang, Chun-Yang

    2016-09-20

    Biosensors for highly sensitive, selective, and rapid quantification of specific biomolecules make great contributions to biomedical research, especially molecular diagnostics. However, conventional methods for biomolecular assays often suffer from insufficient sensitivity and poor specificity. In some case (e.g., early disease diagnostics), the concentration of target biomolecules is too low to be detected by these routine approaches, and cumbersome procedures are needed to improve the detection sensitivity. Therefore, there is an urgent need for rapid and ultrasensitive analytical tools. In this respect, single-molecule fluorescence approaches may well satisfy the requirement and hold promising potential for the development of ultrasensitive biosensors. Encouragingly, owing to the advances in single-molecule microscopy and spectroscopy over past decades, the detection of single fluorescent molecule comes true, greatly boosting the development of highly sensitive biosensors. By in vitro/in vivo labeling of target biomolecules with proper fluorescent tags, the quantification of certain biomolecule at the single-molecule level is achieved. In comparison with conventional ensemble measurements, single-molecule detection-based analytical methods possess the advantages of ultrahigh sensitivity, good selectivity, rapid analysis time, and low sample consumption. Consequently, single-molecule detection may be potentially employed as an ideal analytical approach to quantify low-abundant biomolecules with rapidity and simplicity. In this Account, we will summarize our efforts for developing a series of ultrasensitive biosensors based on single-molecule counting. Single-molecule counting is a member of single-molecule detection technologies and may be used as a very simple and ultrasensitive method to quantify target molecules by simply counting the individual fluorescent bursts. In the fluorescent sensors, the signals of target biomolecules may be translated to the

  2. Fluorescence quenching based alkaline phosphatase activity detection.

    Science.gov (United States)

    Mei, Yaqi; Hu, Qiong; Zhou, Baojing; Zhang, Yonghui; He, Minhui; Xu, Ting; Li, Feng; Kong, Jinming

    2018-01-01

    Simple and fast detection of alkaline phosphatase (ALP) activity is of great importance for diagnostic and analytical applications. In this work, we report a turn-off approach for the real-time detection of ALP activity on the basis of the charge transfer induced fluorescence quenching of the Cu(BCDS) 2 2- (BCDS = bathocuproine disulfonate) probe. Initially, ALP can enzymatically hydrolyze the substrate ascorbic acid 2-phosphate to release ascorbic acid (AA). Subsequently, the AA-mediated reduction of the Cu(BCDS) 2 2- probe, which displays an intense photoluminescence band at the wavelength of 402nm, leads to the static quenching of fluorescence of the probe as a result of charge transfer. The underlying mechanism of the fluorescence quenching was demonstrated by quantum mechanical calculations. The Cu(BCDS) 2 2- probe features a large Stokes shift (86nm) and is highly immune to photo bleaching. In addition, this approach is free of elaborately designed fluorescent probes and allows the detection of ALP activity in a real-time manner. Under optimal conditions, it provides a fast and sensitive detection of ALP activity within the dynamic range of 0-220mUmL -1 , with a detection limit down to 0.27mUmL -1 . Results demonstrate that it is highly selective, and applicable to the screening of ALP inhibitors in drug discovery. More importantly, it shows a good analytical performance for the direct detection of the endogenous ALP levels of undiluted human serum and even whole blood samples. Therefore, the proposed charge transfer based approach has great potential in diagnostic and analytical applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Time-Resolved Fluorescence Anisotropy of Bicyclo[1.1.1]pentane/Tolane-Based Molecular Rods Included in Tris(o-phenylenedioxy)cyclotriphosphazene (TPP)

    Czech Academy of Sciences Publication Activity Database

    Cipolloni, Marco; Kaleta, Jiří; Mašát, Milan; Dron, P. I.; Shen, Y.; Zhao, K.; Rogers, C. T.; Shoemaker, R. K.; Michl, Josef

    2015-01-01

    Roč. 119, č. 16 (2015), s. 8805-8820 ISSN 1932-7447 EU Projects: European Commission(XE) 227756 - DIPOLAR ROTOR ARRAY Institutional support: RVO:61388963 Keywords : compound based approach * stretched polyethylene * solvent invironment Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 4.509, year: 2015 http://pubs.acs.org/doi/pdf/10.1021/acs.jpcc.5b01960

  4. Molecular Viscosity Sensors with Two Rotators for Optimizing the Fluorescence Intensity-Contrast Trade-Off.

    Science.gov (United States)

    Lee, Seung-Chul; Lee, Chang-Lyoul; Heo, Jeongyun; Jeong, Chan-Uk; Lee, Gyeong-Hui; Kim, Sehoon; Yoon, Woojin; Yun, Hoseop; Park, Sung O; Kwak, Sang Kyu; Park, Sung-Ha; Kwon, O-Pil

    2018-02-26

    A series of fluorescent molecular rotors obtained by introducing two rotational groups ("rotators"), which exhibit different rotational and electron-donating abilities, are discussed. Whereas the control molecular rotor, PH, includes a single rotator (the widely used phenyl group), the PO molecular rotors consist of two rotators (a phenyl group and an alkoxy group), which exhibit simultaneous strongly electron-donating and easy rotational abilities. Compared with the control rotor PH, PO molecular rotors exhibited one order of magnitude higher quantum yield (fluorescence intensity) and simultaneously exhibited significantly higher fluorescence contrast. These properties are directly related to the strong electron-donating ability and low energy barrier of rotation of the alkoxy group, as confirmed by dynamic fluorescence experiments and quantum chemical calculations. The PO molecular rotors exhibited two fluorescence relaxation pathways, whereas the PH molecular rotor exhibited a single fluorescence relaxation pathway. Cellular fluorescence imaging with PO molecular rotors for mapping cellular viscosity was successfully demonstrated. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Spectral selective fluorescence molecular imaging with volume holographic imaging system

    Directory of Open Access Journals (Sweden)

    Yanlu Lv

    2016-03-01

    Full Text Available A compact volume holographic imaging (VHI method that can detect fluorescence objects located in diffusive medium in spectral selective imaging manner is presented. The enlargement of lateral field of view of the VHI system is realized by using broadband illumination and demagnification optics. Each target spectrum of fluorescence emitting from a diffusive medium is probed by tuning the inclination angle of the transmission volume holographic grating (VHG. With the use of the single transmission VHG, fluorescence images with different spectrum are obtained sequentially and precise three-dimensional (3D information of deep fluorescent objects located in a diffusive medium can be reconstructed from these images. The results of phantom experiments demonstrate that two fluorescent objects with a sub-millimeter distance can be resolved by spectral selective imaging.

  6. Molecular quantification of genes encoding for green-fluorescent proteins

    DEFF Research Database (Denmark)

    Felske, A; Vandieken, V; Pauling, B V

    2003-01-01

    A quantitative PCR approach is presented to analyze the amount of recombinant green fluorescent protein (gfp) genes in environmental DNA samples. The quantification assay is a combination of specific PCR amplification and temperature gradient gel electrophoresis (TGGE). Gene quantification...

  7. Dual-modality, fluorescent, PLGA encapsulated bismuth nanoparticles for molecular and cellular fluorescence imaging and computed tomography.

    Science.gov (United States)

    Swy, Eric R; Schwartz-Duval, Aaron S; Shuboni, Dorela D; Latourette, Matthew T; Mallet, Christiane L; Parys, Maciej; Cormode, David P; Shapiro, Erik M

    2014-11-07

    Reports of molecular and cellular imaging using computed tomography (CT) are rapidly increasing. Many of these reports use gold nanoparticles. Bismuth has similar CT contrast properties to gold while being approximately 1000-fold less expensive. Herein we report the design, fabrication, characterization, and CT and fluorescence imaging properties of a novel, dual modality, fluorescent, polymer encapsulated bismuth nanoparticle construct for computed tomography and fluorescence imaging. We also report on cellular internalization and preliminary in vitro and in vivo toxicity effects of these constructs. 40 nm bismuth(0) nanocrystals were synthesized and encapsulated within 120 nm Poly(dl-lactic-co-glycolic acid) (PLGA) nanoparticles by oil-in-water emulsion methodologies. Coumarin-6 was co-encapsulated to impart fluorescence. High encapsulation efficiency was achieved ∼70% bismuth w/w. Particles were shown to internalize within cells following incubation in culture. Bismuth nanocrystals and PLGA encapsulated bismuth nanoparticles exhibited >90% and >70% degradation, respectively, within 24 hours in acidic, lysosomal environment mimicking media and both remained nearly 100% stable in cytosolic/extracellular fluid mimicking media. μCT and clinical CT imaging was performed at multiple X-ray tube voltages to measure concentration dependent attenuation rates as well as to establish the ability to detect the nanoparticles in an ex vivo biological sample. Dual fluorescence and CT imaging is demonstrated as well. In vivo toxicity studies in rats revealed neither clinically apparent side effects nor major alterations in serum chemistry and hematology parameters. Calculations on minimal detection requirements for in vivo targeted imaging using these nanoparticles are presented. Indeed, our results indicate that these nanoparticles may serve as a platform for sensitive and specific targeted molecular CT and fluorescence imaging.

  8. Green fluorescent protein with anionic tryptophan-based chromophore and long fluorescence lifetime.

    Science.gov (United States)

    Sarkisyan, Karen S; Goryashchenko, Alexander S; Lidsky, Peter V; Gorbachev, Dmitry A; Bozhanova, Nina G; Gorokhovatsky, Andrey Yu; Pereverzeva, Alina R; Ryumina, Alina P; Zherdeva, Victoria V; Savitsky, Alexander P; Solntsev, Kyril M; Bommarius, Andreas S; Sharonov, George V; Lindquist, Jake R; Drobizhev, Mikhail; Hughes, Thomas E; Rebane, Aleksander; Lukyanov, Konstantin A; Mishin, Alexander S

    2015-07-21

    Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP-the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  9. Separate and simultaneous binding effects of aspirin and amlodipine to human serum albumin based on fluorescence spectroscopic and molecular modeling characterizations: A mechanistic insight for determining usage drugs doses

    International Nuclear Information System (INIS)

    Abdollahpour, Nooshin; Asoodeh, Ahmad; Saberi, Mohammad Reza; Chamani, JamshidKhan

    2011-01-01

    The binding of aspirin (ASA) and amlodipine (AML) to human serum albumin (HSA) in aqueous solution was investigated by multiple techniques such as fluorescence quenching, resonance light scattering (RLS), three-dimensional fluorescence spectroscopy, FT-IR and zeta-potential measurements in an aqueous solution at pH=7.4. For the protein-ligand association reaction, fluorescence measurements can give important clues as to the binding of ligands to proteins, e.g., the binding mechanism, binding mode, binding constants, binding sites, etc. Fluorescence spectroscopy showed that ASA and AML could quench the HSA fluorescence spectra, and this quenching effect became more significant when both ASA and AML coexisted. The results pointed at the interaction between HSA and both drugs as ternary systems decreasing the binding constant and binding stability of the HSA-drug complex as a binary system. Therefore, by reducing the amount of drugs transported to their targets, the free drug concentration of the target would be reduced, lowering the efficacy of the drugs. It was demonstrated that there exists antagonistic behavior between the two drugs when it comes to binding of HSA. Furthermore, the fluorescence results also showed that the quenching mechanism of HSA-drug complexes as binary and ternary systems is a static procedure. The number of binding sites of HSA-ASA, (HSA-AML)ASA, HSA-AML and (HSA-ASA) AML were 1.31, 0.92, 1 and 0.93, respectively. Due to the existence of the antagonistic action between ASA and AML, the binding distance r was reduced. The results of synchronous fluorescence and three-dimensional fluorescence spectra showed that the antagonistic action between ASA and AML would alter the micro-environment around Trp and Tyr residues. Moreover, the simultaneous presence of ASA and AML during binding to HSA should be taken into account in multidrug therapy, as it induces the necessity of a monitoring therapy owing to the possible increase of uncontrolled toxic

  10. Separate and simultaneous binding effects of aspirin and amlodipine to human serum albumin based on fluorescence spectroscopic and molecular modeling characterizations: A mechanistic insight for determining usage drugs doses

    Energy Technology Data Exchange (ETDEWEB)

    Abdollahpour, Nooshin [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of); Asoodeh, Ahmad [Department of Chemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad (Iran, Islamic Republic of); Saberi, Mohammad Reza [Department of Medical Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of); Chamani, JamshidKhan, E-mail: chamani@ibb.ut.ac.ir [Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad (Iran, Islamic Republic of)

    2011-09-15

    The binding of aspirin (ASA) and amlodipine (AML) to human serum albumin (HSA) in aqueous solution was investigated by multiple techniques such as fluorescence quenching, resonance light scattering (RLS), three-dimensional fluorescence spectroscopy, FT-IR and zeta-potential measurements in an aqueous solution at pH=7.4. For the protein-ligand association reaction, fluorescence measurements can give important clues as to the binding of ligands to proteins, e.g., the binding mechanism, binding mode, binding constants, binding sites, etc. Fluorescence spectroscopy showed that ASA and AML could quench the HSA fluorescence spectra, and this quenching effect became more significant when both ASA and AML coexisted. The results pointed at the interaction between HSA and both drugs as ternary systems decreasing the binding constant and binding stability of the HSA-drug complex as a binary system. Therefore, by reducing the amount of drugs transported to their targets, the free drug concentration of the target would be reduced, lowering the efficacy of the drugs. It was demonstrated that there exists antagonistic behavior between the two drugs when it comes to binding of HSA. Furthermore, the fluorescence results also showed that the quenching mechanism of HSA-drug complexes as binary and ternary systems is a static procedure. The number of binding sites of HSA-ASA, (HSA-AML)ASA, HSA-AML and (HSA-ASA) AML were 1.31, 0.92, 1 and 0.93, respectively. Due to the existence of the antagonistic action between ASA and AML, the binding distance r was reduced. The results of synchronous fluorescence and three-dimensional fluorescence spectra showed that the antagonistic action between ASA and AML would alter the micro-environment around Trp and Tyr residues. Moreover, the simultaneous presence of ASA and AML during binding to HSA should be taken into account in multidrug therapy, as it induces the necessity of a monitoring therapy owing to the possible increase of uncontrolled toxic

  11. Polypeptides Based Molecular Electronics

    National Research Council Canada - National Science Library

    Lam, Yeng M; Mhaisalkar, Subodh; Li, Lain-Jong; Dravid, Vinayak P; Shekhawat, Gajendra S; Suri, Raman

    2008-01-01

    ... the formation of molecular devices such as transistors, diodes, and sensors. We have designed the peptides, arranged them on substrates using self-assembly, Dip-PEN nanolithography, and also e-beam assisted lithography...

  12. A study of molecular dynamics within liquid flows using fluorescence depolarization

    Science.gov (United States)

    Kenyon, A. J.; McCaffery, A. J.; Quintella, C. M.; Winkel, J. F.

    Molecular alignment within thin laminar jets of rhodamine 6G in solution in ethylene glycol has been investigated by measuring the depolarization of laser induced fluorescence. From measurements of a large number of points within the flow, we have built up maps of molecular alignment which display in detail the distribution and evolution of flow alignment. On the basis of these observations, we analyse the evolution of molecular alignment in terms of internal shear forces resulting from velocity distributions within liquid flows.

  13. CMOS Time-Resolved, Contact, and Multispectral Fluorescence Imaging for DNA Molecular Diagnostics

    Directory of Open Access Journals (Sweden)

    Nan Guo

    2014-10-01

    Full Text Available Instrumental limitations such as bulkiness and high cost prevent the fluorescence technique from becoming ubiquitous for point-of-care deoxyribonucleic acid (DNA detection and other in-field molecular diagnostics applications. The complimentary metal-oxide-semiconductor (CMOS technology, as benefited from process scaling, provides several advanced capabilities such as high integration density, high-resolution signal processing, and low power consumption, enabling sensitive, integrated, and low-cost fluorescence analytical platforms. In this paper, CMOS time-resolved, contact, and multispectral imaging are reviewed. Recently reported CMOS fluorescence analysis microsystem prototypes are surveyed to highlight the present state of the art.

  14. Molecular imaging with optics: primer and case for near-infrared fluorescence techniques in personalized medicine

    Science.gov (United States)

    Sevick-Muraca, Eva M.; Rasmussen, John C.

    2010-01-01

    We compare and contrast the development of optical molecular imaging techniques with nuclear medicine with a didactic emphasis for initiating readers into the field of molecular imaging. The nuclear imaging techniques of gamma scintigraphy, single-photon emission computed tomography, and positron emission tomography are first briefly reviewed. The molecular optical imaging techniques of bioluminescence and fluorescence using gene reporter/probes and gene reporters are described prior to introducing the governing factors of autofluorescence and excitation light leakage. The use of dual-labeled, near-infrared excitable and radio-labeled agents are described with comparative measurements between planar fluorescence and nuclear molecular imaging. The concept of time-independent and -dependent measurements is described with emphasis on integrating time-dependent measurements made in the frequency domain for 3-D tomography. Finally, we comment on the challenges and progress for translating near-infrared (NIR) molecular imaging agents for personalized medicine. PMID:19021311

  15. Molecular engineering and fluorescence for the detection of toxic cations; Ingenierie moleculaire et fluorescence pour la reconnaissance de cations toxiques

    Energy Technology Data Exchange (ETDEWEB)

    Souchon, V

    2007-11-15

    This work is a part of the 'Toxicologie Nucleaire Environnementale' program which aims at studying the effects on the living of heavy metals or radionuclides involved in nuclear industry. Most particularly, it deals with the design of new fluorescent sensors for the selective detection of Pb{sup 2+}, Cd{sup 2+} and Cs{sup +} in biological media. Several fluorescent calixarenes possessing nitrogen atoms were synthesized and their properties as potential lead sensors were investigated. One of them could be used in experimental conditions close to biological media and new target compounds with amide functional groups were proposed. Many approaches were considered for the design of selective fluorescent sensors for cadmium. On the basis of literature results, many chelating compounds incorporating sulfur atoms were synthesized but showed no significant affinity towards cadmium. On the opposite, compounds functionalized with several pyridine-2'-yl-1,2,3-triazol fluorescent moieties linked to a {beta}-cyclodextrin or a calix[4]arene showed good affinity for cadmium in methanol, but the selectivity was found to be insufficient. In contrast, very satisfying results in terms of both selectivity and sensitivity could be obtained with the commercial calcium sensor Rhod-5N in an aqueous medium at neutral pH. Lastly, micromolar detection limits for the selective detection of caesium were reached in an aqueous medium at neutral pH thanks to a new sulfonated fluorescent calixarene with two appended crown-ethers. An original complexation mechanism was proposed and validated by molecular modelling (DFT). (author)

  16. A Quantitative Fluorescence-Based Lipase Assay

    Directory of Open Access Journals (Sweden)

    Giovanna Lomolino

    2012-01-01

    Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

  17. Synthesis and characterization of citrate-based fluorescent small molecules and biodegradable polymers.

    Science.gov (United States)

    Xie, Zhiwei; Kim, Jimin P; Cai, Qing; Zhang, Yi; Guo, Jinshan; Dhami, Ranjodh S; Li, Li; Kong, Bin; Su, Yixue; Schug, Kevin A; Yang, Jian

    2017-03-01

    Novel citric acid based photoluminescent dyes and biodegradable polymers are synthesized via a facile "one-pot" reaction. A comprehensive understanding of the fluorescence mechanisms of the resulting citric acid-based fluorophores is reported. Two distinct types of fluorophores are identified: a thiozolopyridine family with high quantum yield, long lifetime, and exceptional photostability, and a dioxopyridine family with relatively lower quantum yield, multiple lifetimes, and solvent-dependent band shifting behavior. Applications in molecular labeling and cell imaging were demonstrated. The above discoveries contribute to the field of fluorescence chemistry and have laid a solid foundation for further development of new fluorophores and materials that show promise in a diversity of fluorescence-based applications. Photoluminescent materials are pivotal for fluorescence based imaging, labeling and sensing applications. Understanding their fluorescence mechanism is challenging and imperative. We develop a new class of citric acid-derived fluorescent materials in forms of polymers and small molecular dyes by a one-step solvent free reaction. We discovered two different classes of citric acid-derived fluorophores. A two-ring thiozolopyridine structure demonstrates strong fluorescence and exceptional resistance to photo-bleaching. A one-ring dioxopyridine exhibits relative weak fluorescence but with intriguing excitation and solvent-dependent emission wavelength shifting. Our methodology of synthesizing citric acid-derived fluorophores and the understanding on their luminescence are instrumental to the design and production of a large number of new photoluminescent materials for biological and biomedical applications. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  18. Detection of trace tetracycline in fish via synchronous fluorescence quenching with carbon quantum dots coated with molecularly imprinted silica

    Science.gov (United States)

    Yang, Ji; Lin, Zheng-Zhong; Nur, A.-Zha; Lu, Yan; Wu, Ming-Hui; Zeng, Jun; Chen, Xiao-Mei; Huang, Zhi-Yong

    2018-02-01

    A novel fluorescence-based sensor combining synchronous fluorescence spectroscopy (SFS) with molecularly imprinted polymers (MIPs) was fabricated with reverse microemulsion method. Tetracycline (TC), (3-aminopropyl) triethoxysilane (APTES), tetraethyl orthosilicate (TEOS) and carbon quantum dots (CDs) were used as template, functional monomer, cross-linker and signal sources respectively in the probe preparation. A synchronous fluorescence emission (λem) at 355 nm was observed for the prepared MIP-coated CDs (MIP@CDs) particles when the wavelength interval (Δλ) was set as 70 nm, and the synchronous fluorescence intensity could be rapidly and efficiently quenched by TC based on inner filter effect (IFE). The quenching efficiencies of synchronous fluorescence intensity was linearly fitted with tetracycline (TC) concentrations ranging from 0.1 to 50 μmol L- 1 with a detection limit (DL) of 9 nmol L- 1 (3σ, n = 9). The MIP@CDs was used as a probe to detect TC in fish samples with the recoveries ranging from 98.4% to 103.1% and the relative standard deviation less than 6.0%. The results illustrated that the as-prepared MIP@CDs could be applied to the detection of trace TC in fish samples with rapidity, high sensitivity and accuracy.

  19. Tomato seeds maturity detection system based on chlorophyll fluorescence

    Science.gov (United States)

    Li, Cuiling; Wang, Xiu; Meng, Zhijun

    2016-10-01

    Chlorophyll fluorescence intensity can be used as seed maturity and quality evaluation indicator. Chlorophyll fluorescence intensity of seed coats is tested to judge the level of chlorophyll content in seeds, and further to judge the maturity and quality of seeds. This research developed a detection system of tomato seeds maturity based on chlorophyll fluorescence spectrum technology, the system included an excitation light source unit, a fluorescent signal acquisition unit and a data processing unit. The excitation light source unit consisted of two high power LEDs, two radiators and two constant current power supplies, and it was designed to excite chlorophyll fluorescence of tomato seeds. The fluorescent signal acquisition unit was made up of a fluorescence spectrometer, an optical fiber, an optical fiber scaffolds and a narrowband filter. The data processing unit mainly included a computer. Tomato fruits of green ripe stage, discoloration stage, firm ripe stage and full ripe stage were harvested, and their seeds were collected directly. In this research, the developed tomato seeds maturity testing system was used to collect fluorescence spectrums of tomato seeds of different maturities. Principal component analysis (PCA) method was utilized to reduce the dimension of spectral data and extract principal components, and PCA was combined with linear discriminant analysis (LDA) to establish discriminant model of tomato seeds maturity, the discriminant accuracy was greater than 90%. Research results show that using chlorophyll fluorescence spectrum technology is feasible for seeds maturity detection, and the developed tomato seeds maturity testing system has high detection accuracy.

  20. Comparison of molecular imprinted particles prepared using precipitation polymerization in water and chloroform for fluorescent detection of nitroaromatics

    International Nuclear Information System (INIS)

    Stringer, R. Cody; Gangopadhyay, Shubhra; Grant, Sheila A.

    2011-01-01

    Highlights: → Imprinted polymers prepared using precipitation polymerization. → Comparison of chloroform and water as polymerization solvent. → Imprinted polymer doped with quantum dots for fluorescent sensor. → Fluorescent imprinted polymer used to detect nitroaromatic explosives. → Chloroform is ideal solvent for molecular imprinting of nitroaromatics. - Abstract: A comparative study was conducted to study the effects that two different polymerization solvents would have on the properties of imprinted polymer microparticles prepared using precipitation polymerization. Microparticles prepared in chloroform, which previous results indicated was the optimal solvent for molecular imprinting of nitroaromatic explosive compounds, were compared to water, which was hypothesized to decrease water swelling of the polymer and allow enhanced rebinding of aqueous template. The microparticles were characterized and were integrated into a fluorescence sensing mechanism for detection of nitroaromatic explosive compounds. The performance of the sensing mechanisms was compared to illustrate which polymerization solvent produced optimal imprinted polymer microparticles for detection of nitroaromatic molecules. Results indicated that the structures of microparticles synthesized in chloroform versus water varied greatly. Sensor performance studies showed that the microparticles prepared in chloroform had greater imprinting efficiency and higher template rebinding than those prepared in water. For detection of 2,4,6-trinitrotoluene, the chloroform-based fluorescent microparticles achieved a lower limit of detection of 0.1 μM, as compared to 100 μM for the water-based fluorescent microparticles. Detection limits for 2,4-dinitrotoluene, as well as time response studies, also demonstrated that the chloroform-based particles are more effective for detection of nitroaromatic compounds than water-based particles. These results illustrate that the enhanced chemical properties of

  1. Fluorescence depolarization as a probe of molecular dynamics within liquid jets

    Science.gov (United States)

    Kenyon, A. J.; McCaffery, A. J.; Quintella, C. M.

    Preliminary results are presented from a study of fluorescence depolarization within a thin laminar jet of rhodamine 6 G in ethylene glycol. A large number of polarization measurements taken across the jet have enabled us to build up a detailed polarization map of the liquid flow. Relating the degree of depolarization to molecular alignment caused by the presence of shear forces within the jet, we propose that this method may be used as a sensitive probe of the molecular dynamics of liquid flow.

  2. A novel triazole-based fluorescent chemosensor for Zinc ions

    International Nuclear Information System (INIS)

    Gusev, Alexey N.; Shul’gin, Victor F.; Meshkova, Svetlana B.; Smola, Sergey S.; Linert, Wolfgang

    2014-01-01

    This work describes the preparation of novel triazole-based heterocycles and their properties concerning UV–vis absorption and fluorescence emission with and without the presence of metal ions. It is shown that this compound can be used to determine Zn 2+ ion with high selectivity. Therefore, the new compounds are recommended as possible species for the development of new fluorescent probes for the detection of Zn 2+ cation. - Highlights: • Luminescence spectra of Zn(III) complex using triazole ligands were characterized. • New Triazole based ligands exhibiting fluorescence are reported. • Selectivity concerning Zn-ions is established

  3. A novel triazole-based fluorescent chemosensor for Zinc ions

    Energy Technology Data Exchange (ETDEWEB)

    Gusev, Alexey N., E-mail: galex0330@gmail.com [Taurida National V.I. Vernadsky University, Simferopol 295007 (Russian Federation); Shul’gin, Victor F. [Taurida National V.I. Vernadsky University, Simferopol 295007 (Russian Federation); Meshkova, Svetlana B.; Smola, Sergey S. [A.V. Bogatsky Physico-chemical Institute of the National Academy of Sciences of Ukraine, Odessa 65080 Ukraine (Ukraine); Linert, Wolfgang, E-mail: wolfgang.linert@tuwien.ac.at [Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163, A-1060 Vienna (Austria)

    2014-11-15

    This work describes the preparation of novel triazole-based heterocycles and their properties concerning UV–vis absorption and fluorescence emission with and without the presence of metal ions. It is shown that this compound can be used to determine Zn{sup 2+} ion with high selectivity. Therefore, the new compounds are recommended as possible species for the development of new fluorescent probes for the detection of Zn{sup 2+}cation. - Highlights: • Luminescence spectra of Zn(III) complex using triazole ligands were characterized. • New Triazole based ligands exhibiting fluorescence are reported. • Selectivity concerning Zn-ions is established.

  4. Optimized measurements of separations and angles between intra-molecular fluorescent markers

    DEFF Research Database (Denmark)

    Mortensen, Kim; Sung, Jongmin; Flyvbjerg, Henrik

    2015-01-01

    We demonstrate a novel, yet simple tool for the study of structure and function of biomolecules by extending two-colour co-localization microscopy to fluorescent molecules with fixed orientations and in intra-molecular proximity. From each colour-separated microscope image in a time-lapse movie...

  5. Exploring Fluorescence Antibunching in Solution To Determine the Stoichiometry of Molecular Complexes

    Czech Academy of Sciences Publication Activity Database

    Sýkora, Jan; Kaiser, K.; Gregor, I.; Bönigk, W.; Schmalzing, G.; Enderlain, J.

    2007-01-01

    Roč. 79, - (2007), s. 4040-4049 ISSN 0003-2700 R&D Projects: GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z40400503 Keywords : exploring fluorescence antibunching * molecular complex es * biophysical methods Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 5.287, year: 2007

  6. Confirmation of molecular formulas of metallic complexes through X-ray fluorescence quantitative analysis

    International Nuclear Information System (INIS)

    Filgueiras, C.A.L.; Marques, E.V.; Machado, R.M.

    1984-01-01

    X-ray fluorescence spectrophotometry was employed to determined the metal content in a series of five transition element complexes (Mn, Ti, Zn, V). The results confirmed the molecular formulas of these complexes, already proposed on the basis of elemental microanalysis, solution condutimetry and other analytical methods. (C.L.B.) [pt

  7. Advances in Spiropyrans/Spirooxazines and Applications Based on Fluorescence Resonance Energy Transfer (FRET with Fluorescent Materials

    Directory of Open Access Journals (Sweden)

    Hongyan Xia

    2017-12-01

    Full Text Available Studies on the following were reviewed: (1 the structure of spiropyrans and spirooxazines (two kinds of spiro compounds under external stimuli and (2 the construction and applications of composite systems based on fluorescence resonance energy transfer (FRET with fluorescent materials. When treated with different stimuli (light, acids and bases, solvents, metal ions, temperature, redox potential, and so on, spiropyrans/spirooxazines undergo transformations between the ring-closed form (SP, the ring-opened merocyanine (MC form, and the protonated ring-opened form (MCH. This is due to the breakage of the spiro C–O bond and the protonation of MC, along with a color change. Various novel, multifunctional materials based on photochromic spiropyrans and spirooxazines have been successfully developed because of the vastly differently physiochemical properties posssed by the SP, MC and MCH forms. Among the three different structural forms, the MC form has been studied most extensively. The MC form not only gives complexes with various inorganic particles, biological molecules, and organic chemicals but also acts as the energy acceptor (of energy from fluorescent molecules during energy transfer processes that take place under proper conditions. Furthermore, spiropyran and spirooxazine compounds exhibit reversible physicochemical property changes under proper stimuli; this provides more advantages compared with other photochromic compounds. Additionally, the molecular structures of spiropyrans and spirooxazines can be easily modified and extended, so better compounds can be obtained to expand the scope of already known applications. Described in detail are: (1 the structural properties of spiropyrans and spirooxazines and related photochromic mechanisms; (2 composite systems based on spiropyrans and spirooxazines, and (3 fluorescent materials which have potential applications in sensing, probing, and a variety of optical elements.

  8. AIEgen-Based Fluorescent Nanomaterials: Fabrication and Biological Applications

    Directory of Open Access Journals (Sweden)

    Hui Gao

    2018-02-01

    Full Text Available In recent years, luminogens with the feature of aggregation-induced emission (AIEgen have emerged as advanced luminescent materials for fluorescent nanomaterial preparation. AIEgen-based nanomaterials show enhanced fluorescence efficiency and superior photostability, which thusly offer unique advantages in biological applications. In this review, we will summarize the fabrication methods of AIEgen-based nanomaterials and their applications in in vitro/in vivo imaging, cell tracing, photodynamic therapy and drug delivery, focusing on the recent progress.

  9. Intravascular near-infrared fluorescence molecular imaging of atherosclerosis: toward coronary arterial visualization of biologically high-risk plaques

    Science.gov (United States)

    Calfon, Marcella A.; Vinegoni, Claudio; Ntziachristos, Vasilis; Jaffer, Farouc A.

    2010-01-01

    New imaging methods are urgently needed to identify high-risk atherosclerotic lesions prior to the onset of myocardial infarction, stroke, and ischemic limbs. Molecular imaging offers a new approach to visualize key biological features that characterize high-risk plaques associated with cardiovascular events. While substantial progress has been realized in clinical molecular imaging of plaques in larger arterial vessels (carotid, aorta, iliac), there remains a compelling, unmet need to develop molecular imaging strategies targeted to high-risk plaques in human coronary arteries. We present recent developments in intravascular near-IR fluorescence catheter-based strategies for in vivo detection of plaque inflammation in coronary-sized arteries. In particular, the biological, light transmission, imaging agent, and engineering principles that underlie a new intravascular near-IR fluorescence sensing method are discussed. Intravascular near-IR fluorescence catheters appear highly translatable to the cardiac catheterization laboratory, and thus may offer a new in vivo method to detect high-risk coronary plaques and to assess novel atherosclerosis biologics.

  10. Genome-wide functional analysis on the molecular mechanism of specifically biosynthesized fluorescence Eu complex.

    Science.gov (United States)

    Ye, Jing; Dong, Xiawei; Jiang, Xuerui; Jiang, Hui; Li, Chen-Zhong; Wang, Xuemei

    2017-09-22

    Fluorescence imaging as an attractive diagnostic technique is widely employed for early diagnosis of cancer. Self-biosynthesized fluorescent Eu complex in situ in Hela cells have realized specifically and accurately fluorescence imaging for cancer cells. But the molecular mechanism of the in situ biosynthesized process is still unclear. In order to reveal this mechanism, we have investigated whole-genome expression profiles with cDNA microarray, incubated with Eu solution in Hela cells for 24 h. Methylthiazoltetrazolium (MTT) assay and laser confocal fluorescence microscopy study showed the low cytotoxicity and specifically fluorescence imaging of Eu complex in Hela cells. It is observed that 563 up-regulated genes and 274 down-regulated genes were differentially expressed. Meanwhile, quantitative RT-PCR was utilized to measure the expression of some important genes, which validated the results of microarray data analysis. Besides, GO analysis showed that a wide range of differential expression functional genes involved in three groups, including cellular component, molecular function and cellular biological process. It was evident that some important biological pathways were apparently affected through KEGG pathway analysis, including focal adhesion pathway and PI3K (phosphatidylinositol 3' -kinase)-Akt signaling pathway, which can influence glycolytic metabolism and NAD(P)H-oxidases metabolic pathway.

  11. Synthesis, characterization and fluorescence performance of a waterborne polyurethane-based polymeric dye

    Energy Technology Data Exchange (ETDEWEB)

    Xianhai, Hu, E-mail: hxyh@aiai.edu.cn [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China); School of Materials and Chemical Engineering, Building Energy Efficiency Research Institute, Anhui University of Architecture, Hefei 230022 (China); Zhang, Xingyuan, E-mail: zxym@ustc.edu.cn [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China); Liu, Jin [School of Materials and Chemical Engineering, Building Energy Efficiency Research Institute, Anhui University of Architecture, Hefei 230022 (China); Dai, Jiabing [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China)

    2013-10-15

    A novel anionic waterborne polyurethane-based fluorescent dye WPU-DV26 was synthesized by incorporating the molecular structure of disperse violet 26 (DV26) into the polyurethane chain. The structure of WPU-DV26 was confirmed by means of Fourier transform infrared spectroscopy and UV–vis absorption analysis. Comparing to the UV–vis spectrum of DV26, WPU-DV26 showed a hypsochromic shift from the absorption maxima of 518, 558, 609 nm to 510, 548, 586 nm, respectively. WPU-DV26 can form stable latex in water. The number average molecular weight and its distribution index, and average latex particle size for WPU-DV26 were determined to be 2.33×10{sup 4}, 1.36 and 80 nm, respectively. The improved thermal stability of WPU-DV26 can be attributed to the embedded anthraquinone unit of DV26. It was found that both the intensity and stability of the fluorescence of WPU-DV26 latex were improved significantly compared with those of DV26. -- Highlights: ► A waterborne polyurethane-based polymeric dye was synthesized. ► The fluorescence intensity of WPU-DV26 emulsion was enhanced greatly compared with that of DV26. ► The fluorescence stability of WPU-DV26 emulsion was fine not only for long term storage but also for fluorescence quencher.

  12. Antigen-dependent fluorescence response of anti-c-Myc Quenchbody studied by molecular dynamics simulations

    Science.gov (United States)

    Mori, Yoshiharu; Okumura, Hisashi; Watanabe, Takayoshi; Hohsaka, Takahiro

    2018-04-01

    We performed metadynamics molecular dynamics simulations to reveal mechanism of antigen-dependent fluorescence response observed for site-specifically fluorescent-labeled single-chain antibody against c-Myc peptide antigen. We found that VH and VL bind with each other only when the antigen exists and that the fluorophore labeled at the N-terminus of VH interacts with Trp103 most stably. These results support the mechanism proposed from previous experiments: In the absence of antigen, Trp residues are partially exposed at the interface of VH and quench the fluorophore. In the presence of antigen, the Trp residues are buried between VH and VL , and the quenching is eliminated.

  13. Application of locked nucleic acid-based probes in fluorescence in situ hybridization

    DEFF Research Database (Denmark)

    Fontenete, Sílvia; Carvalho, Daniel R; Guimarães, Nuno

    2016-01-01

    of nucleic acid mimics used as mixmers in LNA-based probes strongly influence the efficiency of detection. LNA probes with 10 to 15 mers showed the highest efficiency. Additionally, the combination of 2′-OMe RNA with LNA allowed an increase on the fluorescence intensities of the probes. Overall......Fluorescence in situ hybridization (FISH) employing nucleic acid mimics as probes is becoming an emerging molecular tool in the microbiology area for the detection and visualization of microorganisms. However, the impact that locked nucleic acid (LNA) and 2′-O-methyl (2′-OMe) RNA modifications have...

  14. Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

    Directory of Open Access Journals (Sweden)

    Davidson Michael W

    2008-03-01

    Full Text Available Abstract Background In the 15 years that have passed since the cloning of Aequorea victoria green fluorescent protein (avGFP, the expanding set of fluorescent protein (FP variants has become entrenched as an indispensable toolkit for cell biology research. One of the latest additions to the toolkit is monomeric teal FP (mTFP1, a bright and photostable FP derived from Clavularia cyan FP. To gain insight into the molecular basis for the blue-shifted fluorescence emission we undertook a mutagenesis-based study of residues in the immediate environment of the chromophore. We also employed site-directed and random mutagenesis in combination with library screening to create new hues of mTFP1-derived variants with wavelength-shifted excitation and emission spectra. Results Our results demonstrate that the protein-chromophore interactions responsible for blue-shifting the absorbance and emission maxima of mTFP1 operate independently of the chromophore structure. This conclusion is supported by the observation that the Tyr67Trp and Tyr67His mutants of mTFP1 retain a blue-shifted fluorescence emission relative to their avGFP counterparts (that is, Tyr66Trp and Tyr66His. Based on previous work with close homologs, His197 and His163 are likely to be the residues with the greatest contribution towards blue-shifting the fluorescence emission. Indeed we have identified the substitutions His163Met and Thr73Ala that abolish or disrupt the interactions of these residues with the chromophore. The mTFP1-Thr73Ala/His163Met double mutant has an emission peak that is 23 nm red-shifted from that of mTFP1 itself. Directed evolution of this double mutant resulted in the development of mWasabi, a new green fluorescing protein that offers certain advantages over enhanced avGFP (EGFP. To assess the usefulness of mTFP1 and mWasabi in live cell imaging applications, we constructed and imaged more than 20 different fusion proteins. Conclusion Based on the results of our

  15. Fluorescence multispectral imaging-based diagnostic system for atherosclerosis.

    Science.gov (United States)

    Ho, Cassandra Su Lyn; Horiuchi, Toshikatsu; Taniguchi, Hiroaki; Umetsu, Araya; Hagisawa, Kohsuke; Iwaya, Keiichi; Nakai, Kanji; Azmi, Amalina; Zulaziz, Natasha; Azhim, Azran; Shinomiya, Nariyoshi; Morimoto, Yuji

    2016-08-20

    Composition of atherosclerotic arterial walls is rich in lipids such as cholesterol, unlike normal arterial walls. In this study, we aimed to utilize this difference to diagnose atherosclerosis via multispectral fluorescence imaging, which allows for identification of fluorescence originating from the substance in the arterial wall. The inner surface of extracted arteries (rabbit abdominal aorta, human coronary artery) was illuminated by 405 nm excitation light and multispectral fluorescence images were obtained. Pathological examination of human coronary artery samples were carried out and thickness of arteries were calculated by measuring combined media and intima thickness. The fluorescence spectra in atherosclerotic sites were different from those in normal sites. Multiple regions of interest (ROI) were selected within each sample and a ratio between two fluorescence intensity differences (where each intensity difference is calculated between an identifier wavelength and a base wavelength) from each ROI was determined, allowing for discrimination of atherosclerotic sites. Fluorescence intensity and thickness of artery were found to be significantly correlated. These results indicate that multispectral fluorescence imaging provides qualitative and quantitative evaluations of atherosclerosis and is therefore a viable method of diagnosing the disease.

  16. Self-prior strategy for organ reconstruction in fluorescence molecular tomography.

    Science.gov (United States)

    Zhou, Yuan; Chen, Maomao; Su, Han; Luo, Jianwen

    2017-10-01

    The purpose of this study is to propose a strategy for organ reconstruction in fluorescence molecular tomography (FMT) without prior information from other imaging modalities, and to overcome the high cost and ionizing radiation caused by the traditional structural prior strategy. The proposed strategy is designed as an iterative architecture to solve the inverse problem of FMT. In each iteration, a short time Fourier transform (STFT) based algorithm is used to extract the self-prior information in the space-frequency energy spectrum with the assumption that the regions with higher fluorescence concentration have larger energy intensity, then the cost function of the inverse problem is modified by the self-prior information, and lastly an iterative Laplacian regularization algorithm is conducted to solve the updated inverse problem and obtains the reconstruction results. Simulations and in vivo experiments on liver reconstruction are carried out to test the performance of the self-prior strategy on organ reconstruction. The organ reconstruction results obtained by the proposed self-prior strategy are closer to the ground truth than those obtained by the iterative Tikhonov regularization (ITKR) method (traditional non-prior strategy). Significant improvements are shown in the evaluation indexes of relative locational error (RLE), relative error (RE) and contrast-to-noise ratio (CNR). The self-prior strategy improves the organ reconstruction results compared with the non-prior strategy and also overcomes the shortcomings of the traditional structural prior strategy. Various applications such as metabolic imaging and pharmacokinetic study can be aided by this strategy.

  17. DNA-Based Self-Assembly of Fluorescent Nanodiamonds.

    Science.gov (United States)

    Zhang, Tao; Neumann, Andre; Lindlau, Jessica; Wu, Yuzhou; Pramanik, Goutam; Naydenov, Boris; Jelezko, Fedor; Schüder, Florian; Huber, Sebastian; Huber, Marinus; Stehr, Florian; Högele, Alexander; Weil, Tanja; Liedl, Tim

    2015-08-12

    As a step toward deterministic and scalable assembly of ordered spin arrays we here demonstrate a bottom-up approach to position fluorescent nanodiamonds (NDs) with nanometer precision on DNA origami structures. We have realized a reliable and broadly applicable surface modification strategy that results in DNA-functionalized and perfectly dispersed NDs that were then self-assembled in predefined geometries. With optical studies we show that the fluorescence properties of the nitrogen-vacancy color centers in NDs are preserved during surface modification and DNA assembly. As this method allows the nanoscale arrangement of fluorescent NDs together with other optically active components in complex geometries, applications based on self-assembled spin lattices or plasmon-enhanced spin sensors as well as improved fluorescent labeling for bioimaging could be envisioned.

  18. Motor oil classification based on time-resolved fluorescence.

    Science.gov (United States)

    Mu, Taotao; Chen, Siying; Zhang, Yinchao; Guo, Pan; Chen, He; Meng, Fandong

    2014-01-01

    A time-resolved fluorescence (TRF) technique is presented for classifying motor oils. The system is constructed with a third harmonic Nd:YAG laser, a spectrometer, and an intensified charge coupled device (ICCD) camera. Steady-state and time-resolved fluorescence (TRF) measurements are reported for several motor oils. It is found that steady-state fluorescence is insufficient to distinguish the motor oil samples. Then contour diagrams of TRF intensities (CDTRFIs) are acquired to serve as unique fingerprints to identify motor oils by using the distinct TRF of motor oils. CDTRFIs are preferable to steady-state fluorescence spectra for classifying different motor oils, making CDTRFIs a particularly choice for the development of fluorescence-based methods for the discrimination and characterization of motor oils. The two-dimensional fluorescence contour diagrams contain more information, not only the changing shapes of the LIF spectra but also the relative intensity. The results indicate that motor oils can be differentiated based on the new proposed method, which provides reliable methods for analyzing and classifying motor oils.

  19. Smart phone based bacterial detection using bio functionalized fluorescent nanoparticles

    International Nuclear Information System (INIS)

    Rajendran, Vinoth Kumar; Bakthavathsalam, Padmavathy; Ali, Baquir Mohammed Jaffar

    2014-01-01

    We are describing immunochromatographic test strips with smart phone-based fluorescence readout. They are intended for use in the detection of the foodborne bacterial pathogens Salmonella spp. and Escherichia coli O157. Silica nanoparticles (SiNPs) were doped with FITC and Ru(bpy), conjugated to the respective antibodies, and then used in a conventional lateral flow immunoassay (LFIA). Fluorescence was recorded by inserting the nitrocellulose strip into a smart phone-based fluorimeter consisting of a light weight (40 g) optical module containing an LED light source, a fluorescence filter set and a lens attached to the integrated camera of the cell phone in order to acquire high-resolution fluorescence images. The images were analysed by exploiting the quick image processing application of the cell phone and enable the detection of pathogens within few minutes. This LFIA is capable of detecting pathogens in concentrations as low as 10 5 cfu mL −1 directly from test samples without pre-enrichment. The detection is one order of magnitude better compared to gold nanoparticle-based LFIAs under similar condition. The successful combination of fluorescent nanoparticle-based pathogen detection by LFIAs with a smart phone-based detection platform has resulted in a portable device with improved diagnosis features and having potential application in diagnostics and environmental monitoring. (author)

  20. A disposable evanescent wave fiber optic sensor coated with a molecularly imprinted polymer as a selective fluorescence probe.

    Science.gov (United States)

    Ton, Xuan-Anh; Acha, Victor; Bonomi, Paolo; Tse Sum Bui, Bernadette; Haupt, Karsten

    2015-02-15

    We have developed a disposable evanescent wave fiber optic sensor by coating a molecularly imprinted polymer (MIP) containing a fluorescent signaling group on a 4-cm long polystyrene optical waveguide. The MIP is composed of a naphthalimide-based fluorescent monomer, which shows fluorescence enhancement upon binding with carboxyl-containing molecules. The herbicide 2,4-dichlorophenoxyacetic acid and the mycotoxin citrinin were used as model analytes. The coating of the MIP was either performed ex-situ, by dip-coating the fiber with MIP particles synthesized beforehand, or in-situ by evanescent-wave photopolymerization on the fiber. The sensing element was interrogated with a fiber-coupled spectrofluorimeter. The fiber optic sensor detects targets in the low nM range and exhibits specific and selective recognition over structural analogs and non-related carboxyl-containing molecules. This technology can be extended to other carboxyl-containing analytes, and to a broader spectrum of targets using different fluorescent monomers. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Tumor Endothelial Marker Imaging in Melanomas Using Dual-Tracer Fluorescence Molecular Imaging

    Science.gov (United States)

    Tichauer, Kenneth M.; Deharvengt, Sophie J.; Samkoe, Kimberley S.; Gunn, Jason R.; Bosenberg, Marcus W.; Turk, Mary-Jo; Hasan, Tayyaba; Stan, Radu V.; Pogue, Brian W.

    2014-01-01

    Purpose Cancer-specific endothelial markers available for intravascular binding are promising targets for new molecular therapies. In this study, a molecular imaging approach of quantifying endothelial marker concentrations (EMCI) is developed and tested in highly light-absorbing melanomas. The approach involves injection of targeted imaging tracer in conjunction with an untargeted tracer, which is used to account for nonspecific uptake and tissue optical property effects on measured targeted tracer concentrations. Procedures Theoretical simulations and a mouse melanoma model experiment were used to test out the EMCI approach. The tracers used in the melanoma experiments were fluorescently labeled anti-Plvap/PV1 antibody (plasmalemma vesicle associated protein Plvap/PV1 is a transmembrane protein marker exposed on the luminal surface of endothelial cells in tumor vasculature) and a fluorescent isotype control antibody, the uptakes of which were measured on a planar fluorescence imaging system. Results The EMCI model was found to be robust to experimental noise under reversible and irreversible binding conditions and was capable of predicting expected overexpression of PV1 in melanomas compared to healthy skin despite a 5-time higher measured fluorescence in healthy skin compared to melanoma: attributable to substantial light attenuation from melanin in the tumors. Conclusions This study demonstrates the potential of EMCI to quantify endothelial marker concentrations in vivo, an accomplishment that is currently unavailable through any other methods, either in vivo or ex vivo. PMID:24217944

  2. Comparative Phenotypical and Molecular Analyses of Arabidopsis Grown under Fluorescent and LED Light

    OpenAIRE

    Seiler, Franka; Soll, J?rgen; B?lter, Bettina

    2017-01-01

    Comparative analyses of phenotypic and molecular traits of Arabidopsis thaliana grown under standardised conditions is still a challenge using climatic devices supplied with common light sources. These are in most cases fluorescent lights, which have several disadvantages such as heat production at higher light intensities, an invariable spectral output, and relatively rapid “ageing”. This results in non-desired variations of growth conditions and lowers the comparability of data acquired ove...

  3. Synthesis of molecularly imprinted dye-silica nanocomposites with high selectivity and sensitivity: Fluorescent imprinted sensor for rapid and efficient detection of τ-fluvalinate in vodka.

    Science.gov (United States)

    Wang, Yunyun; Wang, Jixiang; Cheng, Rujia; Sun, Lin; Dai, Xiaohui; Yan, Yongsheng

    2018-02-01

    An imprinted fluorescent sensor was fabricated based on SiO 2 nanoparticles encapsulated with a molecularly imprinted polymer containing allyl fluorescein. High fluorine cypermethirin as template molecules, methyl methacrylate as functional monomer, and allyl fluorescein as optical materials synthesized a core-shell fluorescent molecular imprinted sensor, which showed a high and rapid sensitivity and selectivity for the detection of τ-fluvalinate. The sensor presented appreciable sensitivity with a limit of 13.251 nM, rapid detection that reached to equilibrium within 3 min, great linear relationship in the relevant concentration range from 0 to 150 nM, and excellent selectivity over structural analogues. In addition, the fluorescent sensor demonstrated desirable regeneration ability (eight cycling operations). The molecularly imprinted polymers ensured specificity, while the fluorescent dyes provided the stabile sensitivity. Finally, an effective application of the sensor was implemented by the detection of τ-fluvalinate in real samples from vodka. The molecularly imprinted fluorescent sensor showed a promising potential in environmental monitoring and food safety. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Design, synthesis and photochemical properties of the first examples of iminosugar clusters based on fluorescent cores

    Directory of Open Access Journals (Sweden)

    Mathieu L. Lepage

    2015-05-01

    Full Text Available The synthesis and photophysical properties of the first examples of iminosugar clusters based on a BODIPY or a pyrene core are reported. The tri- and tetravalent systems designed as molecular probes and synthesized by way of Cu(I-catalysed azide–alkyne cycloadditions are fluorescent analogues of potent pharmacological chaperones/correctors recently reported in the field of Gaucher disease and cystic fibrosis, two rare genetic diseases caused by protein misfolding.

  5. Chromophore Structure of Photochromic Fluorescent Protein Dronpa: Acid-Base Equilibrium of Two Cis Configurations.

    Science.gov (United States)

    Higashino, Asuka; Mizuno, Misao; Mizutani, Yasuhisa

    2016-04-07

    Dronpa is a novel photochromic fluorescent protein that exhibits fast response to light. The present article is the first report of the resonance and preresonance Raman spectra of Dronpa. We used the intensity and frequency of Raman bands to determine the structure of the Dronpa chromophore in two thermally stable photochromic states. The acid-base equilibrium in one photochromic state was observed by spectroscopic pH titration. The Raman spectra revealed that the chromophore in this state shows a protonation/deprotonation transition with a pKa of 5.2 ± 0.3 and maintains the cis configuration. The observed resonance Raman bands showed that the other photochromic state of the chromophore is in a trans configuration. The results demonstrate that Raman bands selectively enhanced for the chromophore yield valuable information on the molecular structure of the chromophore in photochromic fluorescent proteins after careful elimination of the fluorescence background.

  6. Fluorescence Quantum Yield Measurements of Fluorescent Proteins: A Laboratory Experiment for a Biochemistry or Molecular Biophysics Laboratory Course

    Science.gov (United States)

    Wall, Kathryn P.; Dillon, Rebecca; Knowles, Michelle K.

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts…

  7. Molecular Detection of Bladder Cancer by Fluorescence Microsatellite Analysis and an Automated Genetic Analyzing System

    Directory of Open Access Journals (Sweden)

    Sarel Halachmi

    2007-01-01

    Full Text Available To investigate the ability of an automated fluorescent analyzing system to detect microsatellite alterations, in patients with bladder cancer. We investigated 11 with pathology proven bladder Transitional Cell Carcinoma (TCC for microsatellite alterations in blood, urine, and tumor biopsies. DNA was prepared by standard methods from blood, urine and resected tumor specimens, and was used for microsatellite analysis. After the primers were fluorescent labeled, amplification of the DNA was performed with PCR. The PCR products were placed into the automated genetic analyser (ABI Prism 310, Perkin Elmer, USA and were subjected to fluorescent scanning with argon ion laser beams. The fluorescent signal intensity measured by the genetic analyzer measured the product size in terms of base pairs. We found loss of heterozygocity (LOH or microsatellite alterations (a loss or gain of nucleotides, which alter the original normal locus size in all the patients by using fluorescent microsatellite analysis and an automated analyzing system. In each case the genetic changes found in urine samples were identical to those found in the resected tumor sample. The studies demonstrated the ability to detect bladder tumor non-invasively by fluorescent microsatellite analysis of urine samples. Our study supports the worldwide trend for the search of non-invasive methods to detect bladder cancer. We have overcome major obstacles that prevented the clinical use of an experimental system. With our new tested system microsatellite analysis can be done cheaper, faster, easier and with higher scientific accuracy.

  8. Simultaneous determination of glycols based on fluorescence anisotropy

    International Nuclear Information System (INIS)

    Garcia Sanchez, F.; Navas Diaz, A.; Lopez Guerrero, M.M.

    2007-01-01

    Simultaneous determination of non-fluorescent glycols in mixtures without separation or chemical transformation steps is described. Two methods based in the measure of fluorescence anisotropy of a probe such as fluorescein dissolved in the analyte or analyte mixtures are described. In the first method, the anisotropy spectra of pure and mixtures of analytes are used to quantitative determination (if the fluorophor concentration is in a range where fluorescence intensity is proportional to concentration). In the second method, a calibration curve anisotropy-concentration based on the application of the Perrin equation is established. The methods presented here are capable of directly resolving binary mixtures of non-fluorescent glycols on the basis of differences on the fluorescence anisotropy of a fluorescence tracer. Best analytical performances were obtained by application of the method based on Perrin equation. This method is simple, rapid and allows the determination of mixtures of glycols with reasonable accuracy and precision. Detection limits are limited by the quantum yield and anisotropy values of the tracer in the solvents. Recovery values are related to the differences in anisotropy values of the tracer in the pure solvents. Mixtures of glycerine/ethylene glycol (GL/EG), ethylene glycol/1,2-propane diol (EG/1,2-PPD) and polyethylene glycol 400/1,2-propane diol (PEG 400/1,2-PPD) were analysed and recovery values are within 95-120% in the Perrin method. Relative standard deviation are in the range 1.3-2.9% and detection limits in the range 3.9-8.9%

  9. Fluorescence-based Broad Dynamic Range Viscosity Probes.

    Science.gov (United States)

    Dragan, Anatoliy; Graham, August E; Geddes, Chris D

    2014-03-01

    We introduce two new fluorescent viscosity probes, SYBR Green (SG) and PicoGreen (PG), that we have studied over a broad range of viscosity and in collagen solutions. In water, both dyes have low quantum yields and excited state lifetimes, while in viscous solvents or in complex with DNA both parameters dramatically (300-1000-fold) increase. We show that in log-log scale the dependence of the dyes' quantum yield vs. viscosity is linear, the slope of which is sensitive to temperature. Application of SG and PG, as a fluorescence-based broad dynamic range viscosity probes, to the life sciences is discussed.

  10. Multiplex polymerase chain reaction (PCR) and fluorescence-based ...

    African Journals Online (AJOL)

    Multiplex polymerase chain reaction (PCR) and fluorescence-based capillary electrophoresis (CE) of blood and tissue samples have been used to distinguish between deer species such as red deer, sika deer, wapiti and reindeer. We constructed 4 species-specific primers by using the D-loop of mitochondrial DNA and ...

  11. Fluorescent naphthalene-based benzene tripod for selective ...

    Indian Academy of Sciences (India)

    Aluminium complex of a naphthalene-based benzene tripod ligand system has been reported for the selective recognition of fluoride in aqueous medium in physiological condition. The ligand can selectively recognize Al3+ through enhancement in the fluorescence intensity and this in situ formed aluminium complex ...

  12. A quinoline based pH sensitive ratiometric fluorescent sensor ...

    Indian Academy of Sciences (India)

    Abstract. A new quinoline based hydrazone was synthesized via a condensation reaction and characterized by NMR, mass and single crystal X-ray diffraction studies. It was investigated for suitability as a reversible ratiometric fluorescent pH sensor in acidic pH region. The sensor exhibits intramolecular charge transfer (ICT).

  13. A quinoline based pH sensitive ratiometric fluorescent sensor ...

    Indian Academy of Sciences (India)

    A new quinoline based hydrazone was synthesized via a condensation reaction and characterized by NMR, mass and single crystal X-ray diffraction studies. It was investigated for suitability as a reversible ratiometric fluorescent pH sensor in acidic pH region. The sensor exhibits intramolecular charge transfer (ICT) type ...

  14. Cell-based lipid flippase assay employing fluorescent lipid derivatives

    DEFF Research Database (Denmark)

    Jensen, Maria Stumph; Costa, Sara; Günther-Pomorski, Thomas

    2016-01-01

    , studies of individual P4-ATPase family members from fungi, plants, and animals show that P4-ATPases differ in their substrate specificities and mediate transport of a broader range of lipid substrates. Here, we describe an assay based on fluorescent lipid derivatives to monitor and characterize lipid...

  15. Molecular Dynamic Indicators of the Photoswitching Properties of Green Fluorescent Proteins.

    Science.gov (United States)

    Smyrnova, Daryna; Moeyaert, Benjamien; Michielssens, Servaas; Hofkens, Johan; Dedecker, Peter; Ceulemans, Arnout

    2015-09-10

    Reversibly photoswitchable fluorescent proteins (RSFPs) are highly useful probes for a range of applications including diffraction-unlimited fluorescence microscopy. It was previously shown that reversible photoswitching not only involves cis-trans isomerization and protonation-deprotonation of the chromophore but also results in a marked difference in β-barrel flexibility. In this work, we performed flexibility profiling and functional mode analysis (FMA) using molecular dynamics calculations to study how the flexibility of the RSFP β-barrel influences the photoswitching properties of several fluorescent proteins. We also used Partial Least-Squared (PLS) FMA to detect promising mutation sites for the modulation of photoswitching properties of RSFPs. Our results show that the flexibility of RSFP does depend on its state with a systematically higher flexibility in the dark state compared to the bright state. In particular our method highlights the importance of Val157 in Dronpa, which upon mutation yields a striking difference in the collective motions of the two mutants. Overall, we show that PLS-FMA yields information, complementary to static structures, that can guide the rational design of fluorescent proteins.

  16. Chiral alkylated-aniline as a noninvasive fluorescence sensor: Spectroscopic and molecular modeling studies

    Science.gov (United States)

    Sengupta, Bidisha; Mukherjee, Chirantan Sen; Chakraborty, Sandipan; Muhammad, Maria Jones; Gladney, William; Armstrong, George

    2017-12-01

    Aniline, heterocyclic aromatic amines, and arylamines are known carcinogens. Recently aniline mustard has come into prominence as a novel anticancer agent. In this project, microwave irradiation has been used to synthesize an optically active alkylated aniline namely 2,6-dimethyl-4-(1-(p-tolyl)ethyl)aniline (abbreviated DMPA). The presence of quartet and doublet peaks in NMR and a single chromatogram in HPLC verified that the final product DMPA, prepared from the synthesis reactions, had no major impurities. By using a Lux chiral column in HPLC, two peaks have been detected in the chromatogram, which correspond to two enantiomers of the chiral aniline derivative. Fluorescence spectroscopic measurements on DMPA indicated conspicuous dependence of its emission behavior on the polarity (in terms of the empirical polarity parameter ET(30)) of the homogeneous solvents used, a property important for an optical sensor. The nature of the emission profiles, along with the relevant parameter namely wavelength at emission maximum (λemmax) is used to infer the distribution, binding and microenvironment of the DMPA molecules in human serum albumin protein (HSA). DMPA is weakly fluorescent in aqueous buffer medium, with a dramatic enhancement in the fluorescence emission in the presence of HSA. Molecular modeling studies have been carried out on the two enantiomers (R and S) of DMPA with HSA. The implications of these findings are examined in relation to the potentialities of DMPA as a novel fluorescence sensor for biological systems.

  17. Extensively Reversible Thermal Transformations of a Bistable, Fluorescence-Switchable Molecular Solid: Entry into Functional Molecular Phase-Change Materials.

    Science.gov (United States)

    Srujana, P; Radhakrishnan, T P

    2015-06-15

    Functional phase-change materials (PCMs) are conspicuously absent among molecular materials in which the various attributes of inorganic solids have been realized. While organic PCMs are primarily limited to thermal storage systems, the amorphous-crystalline transformation of materials like Ge-Sb-Te find use in advanced applications such as information storage. Reversible amorphous-crystalline transformations in molecular solids require a subtle balance between robust supramolecular assembly and flexible structural elements. We report novel diaminodicyanoquinodimethanes that achieve this transformation by interlinked helical assemblies coupled with conformationally flexible alkoxyalkyl chains. They exhibit highly reversible thermal transformations between bistable (crystalline/amorphous) forms, along with a prominent switching of the fluorescence emission energy and intensity. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Multimodal fluorescence molecular imaging for in vivo characterization of skin cancer using endogenous and exogenous fluorophores

    Science.gov (United States)

    Miller, Jessica P.; Habimana-Griffin, LeMoyne; Edwards, Tracy S.; Achilefu, Samuel

    2017-06-01

    Similarity of skin cancer with many benign skin pathologies requires reliable methods to detect and differentiate the different types of these lesions. Previous studies have explored the use of disparate optical techniques to identify and estimate the invasive nature of melanoma and basal cell carcinoma with varying outcomes. Here, we used a concerted approach that provides complementary information for rapid screening and characterization of tumors, focusing on squamous cell carcinoma (SCC) of the skin. Assessment of in vivo autofluorescence lifetime (FLT) imaging of endogenous fluorophores that are excitable at longer wavelengths (480 nm) than conventional NADH and FAD revealed a decrease in the short FLT component for SCC compared to normal skin, with mean values of 0.57±0.026 ns and 0.61±0.021 ns, respectively (p=0.004). Subsequent systemic administration of a near-infrared fluorescent molecular probe in SCC bearing mice, followed by the implementation of image processing methods on data acquired from two-dimensional and three-dimensional fluorescence molecular imaging, allowed us to estimate the tumor volume and depth, as well as quantify the fluorescent probe in the tumor. The result suggests the involvement of lipofuscin-like lipopigments and riboflavin in SCC metabolism and serves as a model for staging SCC.

  19. Caffeine and sulfadiazine interact differently with human serum albumin: A combined fluorescence and molecular docking study

    Science.gov (United States)

    Islam, Mullah Muhaiminul; Sonu, Vikash K.; Gashnga, Pynsakhiat Miki; Moyon, N. Shaemningwar; Mitra, Sivaprasad

    2016-01-01

    The interaction and binding behavior of the well-known drug sulfadiazine (SDZ) and psychoactive stimulant caffeine (CAF) with human serum albumin (HSA) was monitored by in vitro fluorescence titration and molecular docking calculations under physiological condition. The quenching of protein fluorescence on addition of CAF is due to the formation of protein-drug complex in the ground state; whereas in case of SDZ, the experimental results were explained on the basis of sphere of action model. Although both these compounds bind preferentially in Sudlow's site 1 of the protein, the association constant is approximately two fold higher in case of SDZ (∼4.0 × 104 M-1) in comparison with CAF (∼9.3 × 102 M-1) and correlates well with physico-chemical properties like pKa and lipophilicity of the drugs. Temperature dependent fluorescence study reveals that both SDZ and CAF bind spontaneously with HSA. However, the binding of SDZ with the protein is mainly governed by the hydrophobic forces in contrast with that of CAF; where, the interaction is best explained in terms of electrostatic mechanism. Molecular docking calculation predicts the binding of these drugs in different location of sub-domain IIA in the protein structure.

  20. A miniaturized cell-based fluorescence resonance energy transfer assay for insulin-receptor activation.

    Science.gov (United States)

    Marine, Shane; Zamiara, Elize; Smith, S Todd; Stec, Erica M; McGarvey, Jeremy; Kornienko, Oleg; Jiang, Guoqiang; Wong, Kenny K; Stack, Jeffrey H; Zhang, Bei B; Ferrer, Marc; Strulovici, Berta

    2006-08-15

    This report describes the development, optimization, and implementation of a miniaturized cell-based assay for the identification of small-molecule insulin mimetics and potentiators. Cell-based assays are attractive formats for compound screening because they present the molecular targets in their cellular environment. A fluorescence resonance energy transfer (FRET) cell-based assay that measures the insulin-dependent colocalization of Akt2 fused with either cyan fluorescent protein or yellow fluorescent protein to the cellular membrane was developed. This ratiometric FRET assay was miniaturized into a robust, yet sensitive 3456-well nanoplate assay with Z' factors of approximately 0.6 despite a very small assay window (less than twofold full activation with insulin). The FRET assay was used for primary screening of a large compound collection for insulin-receptor agonists and potentiators. To prioritize compounds for further development, primary hits were tested in two additional assays, a biochemical time-resolved fluorescence resonance energy transfer assay to measure insulin-receptor phosphorylation and a translocation-based imaging assay. Results from the three assays were combined to yield 11 compounds as potential leads for the development of insulin mimetics or potentiators.

  1. The one-sample PARAFAC approach reveals molecular size distributions of fluorescent components in dissolved organic matter

    DEFF Research Database (Denmark)

    Wünsch, Urban; Murphy, Kathleen R.; Stedmon, Colin

    2017-01-01

    Molecular size plays an important role in dissolved organic matter (DOM) biogeochemistry, but its relationship with the fluorescent fraction of DOM (FDOM) remains poorly resolved. Here high-performance size exclusion chromatography (HPSEC) was coupled to fluorescence emission-excitation (EEM......) spectroscopy in full spectral (60 emission and 34 excitation wavelengths) and chromatographic resolution (... distributions for individual fluorescence components obtained from independent data sets. Spectra extracted from allochthonous DOM were highly similar. Allochthonous and autochthonous DOM shared some spectra, but included unique components. In agreement with the supramolecular assembly hypothesis, molecular...

  2. Binding between Saikosaponin C and Human Serum Albumin by Fluorescence Spectroscopy and Molecular Docking

    Directory of Open Access Journals (Sweden)

    Yi-Cun Chen

    2016-01-01

    Full Text Available Saikosaponin C (SSC is one of the major active constituents of dried Radix bupleuri root (Chaihu in Chinese that has been widely used in China to treat a variety of conditions, such as liver disease, for many centuries. The binding of SSC to human serum albumin (HSA was explored by fluorescence, circular dichroism (CD, UV-vis spectrophotometry, and molecular docking to understand both the pharmacology and the basis of the clinical use of SSC/Chaihu. SSC produced a concentration-dependent quenching effect on the intrinsic fluorescence of HSA, accompanied by a blue shift in the fluorescence spectra. The Stern-Volmer equation showed that this quenching was dominated by static quenching. The binding constant of SSC with HSA was 3.72 × 103 and 2.99 × 103 L·mol−1 at 26 °C and 36 °C, respectively, with a single binding site on each SSC and HSA molecule. Site competitive experiments demonstrated that SSC bound to site I (subdomain IIA and site II (subdomain IIIA in HSA. Analysis of thermodynamic parameters indicated that hydrogen bonding and van der Waals forces were mostly responsible for SSC-HSA association. The energy transfer efficiency and binding distance between SSC and HSA was calculated to be 0.23 J and 2.61 nm at 26 °C, respectively. Synchronous fluorescence and CD measurements indicated that SSC affected HSA conformation in the SSC-HSA complex. Molecular docking supported the experimental findings in conformational changes, binding sites and binding forces, and revealed binding of SSC at the interface between subdomains IIA-IIB.

  3. Photolysis study of octyl p-methoxycinnamate loaded microemulsion by molecular fluorescence and chemometric approach

    Science.gov (United States)

    Nascimento, Danielle Silva; Insausti, Matías; Band, Beatriz Susana Fernández; Grünhut, Marcos

    2018-02-01

    Octyl p-methoxycinnamate (OMC) is one of the most widely used sunscreen agents. However, the efficiency of OMC as UV filter over time is affected due to the formation of the cis-isomer which presents a markedly lower extinction coefficient (εcis = 12,600 L mol- 1 cm- 1 at 291 nm) than the original trans-isomer (εtrans = 24,000 L mol- 1 cm- 1 at 310 nm). In this work, a novel carrier for OMC based on an oil-in-water microemulsion is proposed in order to improve the photostability of this sunscreen. The formulation was composed of 29.2% (w/w) of a 3:1 mixture of ethanol (co-surfactant) and decaethylene glycol mono-dodecyl ether (surfactant), 1.5% (w/w) of oleic acid (oil phase) and 69.2% (w/w) of water. This microemulsion was prepared in a simple way, under moderate stirring at 25 °C and using acceptable, biocompatible and accessible materials for topical use. OMC was incorporated in the vehicle at a final concentration of 5.0% (w/w), taking into account the maximum permitted levels established by international norms. Then, a photolysis study of the loaded formulation was performed using a continuous flow system. The direct photolysis was monitored over time by molecular fluorescence. The recorded spectra data between 370 y 490 nm were analyzed by multivariate curve resolution-alternating least squares algorithm. The kinetic rate constants corresponding to the photolysis of the trans-OMC were calculated from the concentration profiles, resulting in 0.0049 s- 1 for the trans-OMC loaded microemulsion and 0.0131 s- 1 for the trans-OMC in aqueous media. These results demonstrate a higher photostability of the trans-OMC when loaded in the proposed vehicle with respect to the free trans-OMC in aqueous media.

  4. The potential of L-shell X-ray fluorescence CT (XFCT) for molecular imaging.

    Science.gov (United States)

    Bazalova-Carter, Magdalena

    2015-01-01

    X-ray fluorescence CT (XFCT), a novel modality proposed for high-sensitivity high-resolution molecular imaging of probes labelled with a high atomic-number element, has been performed with high-energy K-shell X-rays. XFCT performed with low-energy L-shell X-rays could, in principle, result in an increase of XFCT imaging sensitivity; however, the significant L-shell X-ray attenuation limits its use for imaging of small objects. This commentary discusses the advantages and drawbacks of L-shell XFCT imaging.

  5. Glass-based fluorescence reference materials used for optical and biophotonic applications

    Science.gov (United States)

    Engel, A.; Ottermann, C.; Resch-Genger, U.; Hoffmann, K.; Schweizer, S.; Selling, J.; Spaeth, J.-M.; Rupertus, V.

    2006-04-01

    Fluorescence techniques are known for their high sensitivity and are widely used as analytical tools and detection methods for product and process control, material sciences, environmental and biotechnical analysis, molecular genetics, cell biology, medical diagnostics, and drug screening. For routine measurements by fluorescence techniques the existence of an improved quality assurance is one of the basic needs. According to DIN/ISO 17025 certified standards are used for fluorescence diagnostics having the drawback of giving relative values only. Typical requirements onto fluorescence reference materials or standards deal with the verification of the instrument performance as well as the improvement of the data comparability. Especially for biomedical applications fluorescence labels are used for the detection of proteins. In particular these labels consist of nano crystalline materials like CdS and CdSe. The field of Non-Cadmium containing materials is under investigation. In order to evaluate whether glass based materials can be used as standards it is necessary to calculate absolute values like absorption/excitation cross sections or relative quantum yields. This can be done using different quantities of dopands in glass, glass ceramics or crystals. The investigated materials are based on different types of glass, silicate, phosphate and boron glass, which play a dominant role for the absorption and emission mechanism. Additional to the so-called elementary fluorescence properties induced by raw earth elements the formation of defects lead to higher cross sections additionally. The main investigations deal with wavelength accuracy and lifetime of doped glasses, glass ceramics and crystalline samples. Moreover intensity patterns, homogeneity aspects and photo stability will be discussed.

  6. Development of fluorescent FeIII sensor based on chalcone

    Energy Technology Data Exchange (ETDEWEB)

    Wei Yanli, E-mail: weiyanli@sxu.edu.cn [Research Center of Environmental Science and Engineering, Shanxi University, Taiyuan 030006 (China); Qin Guojie [Institute of Horticulture, Shanxi Academy of Agriculture Science, Taiyuan 030031 (China); Wang Wenyan; Bian Wei; Shuang Shaomin [Research Center of Environmental Science and Engineering, Shanxi University, Taiyuan 030006 (China); Dong Chuan, E-mail: dc@sxu.edu.cn [Research Center of Environmental Science and Engineering, Shanxi University, Taiyuan 030006 (China)

    2011-08-15

    In this paper, 4-dimethylamino 2,5-dihydroxy chalcone (DMADHC), which exhibits excited state intramolecular charge transfer (ICT) characteristics, was synthesized and characterized. A sensitive optochemical sensor for Fe{sup 3+} ion was developed using DMADHC as fluorescence receptor. The fluorescence of DMADHC was gradually quenched with the addition of Fe{sup 3+} ion, which attributed to the formation of 1:1 complex between DMADHC and Fe{sup 3+} ion. The sensor exhibited excellent selectivity for Fe{sup 3+} ion over a large number of cation ions such as alkali, alkaline earth and transitional metal ions with a linear range of 3.984x10{sup -7}-1.135x10{sup -5} and a limit of detection of 8.223x10{sup -8} mol/L. On this basis, the sensor was preliminary applied to the determination of the content of iron ions in multi-vitamin tablet with satisfied results and the recoveries were in the 95-100% interval, and precision (n=5) was better than 5%. - Highlights: > A fluorescence receptor, 4-dimethylamino 2,5-dihydroxy chalcone was synthesis by one-step reaction. > Its intramolecular charge transfer fluorescence characteristics could be blocked by Fe{sup 3+} ion. > Based on this, an optochemical sensor for Fe{sup 3+} ion was developed. > Importantly, our proposed method is particularly useful for determination of Fe{sup 3+} ion in real sample.

  7. A Transformer Partial Discharge Measurement System Based on Fluorescent Fiber

    Directory of Open Access Journals (Sweden)

    Fan Liu

    2012-05-01

    Full Text Available Based on the physical phenomena of optical effects produced by the partial discharge (PD and on the characteristics of fluorescent fiber sensing of weak fluorescent signals, a PD measurement system using a fluorescent fiber sensor was designed. The main parameters of the sensing system were calculated, an experimental testing platform for PD simulation in the lab was established, and PD signals were then detected through ultra-high frequency (UHF and optical methods under a needle-plate discharge model. PD optical pulses in transformer oil contained signal-peak and multi-peak pulse waveforms. Compared with UHF detection results, the number of PD pulses and the elapsed PD pulse phase time revealed a good corresponding relationship. However, PD signal amplitudes presented the opposite, thus indicating that PD UHF signals reflected pulse amplitude value, whereas PD optical signals reflected pulse energy magnitude. The n-u-φ three-dimensional distributions indicated that most of the PD signals concentrated in the nearby industrial frequency voltage peak value. Overall, the proposed fluorescent fiber sensing system design can be used successfully in transformer PD signal detection.

  8. Temperature dependence of the anisotropy of fluorescence in ring molecular systems

    International Nuclear Information System (INIS)

    Herman, Pavel; Barvik, Ivan

    2007-01-01

    The time dependence of the anisotropy of fluorescence after an impulsive excitation in the molecular ring (resembling the B850 ring of the purple bacterium Rhodopseudomonas acidophila) is calculated. Fast fluctuations of the environment are simulated by dynamic disorder and slow fluctuations by uncorrelated static disorder. Without dynamic disorder modest degrees of static disorder are sufficient to cause the experimentally found initial drop of the anisotropy on a sub-100 fs time scale. In the present investigation we are comparing results for the time-dependent optical anisotropy of the molecular ring for four models of the uncorrelated static disorder: Gaussian disorder in the local energies (model A), Gaussian disorder in the transfer integrals (model B), Gaussian disorder in radial positions of molecules (model C) and Gaussian disorder in angular positions of molecules (model D). Both types of disorder-static and dynamic-are taken into account simultaneously

  9. Fluorescence in situ hybridization and molecular studies in infertile men with dysplasia of the fibrous sheath.

    Science.gov (United States)

    Baccetti, Baccio; Collodel, Giulia; Gambera, Laura; Moretti, Elena; Serafini, Francesca; Piomboni, Paola

    2005-07-01

    To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS). Retrospective study. Regional Referral Center for Male Infertility, Siena, Italy. Twelve infertile patients with DFS sperm defects. Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis. The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction. The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls. These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.

  10. Comparative Phenotypical and Molecular Analyses of Arabidopsis Grown under Fluorescent and LED Light.

    Science.gov (United States)

    Seiler, Franka; Soll, Jürgen; Bölter, Bettina

    2017-06-13

    Comparative analyses of phenotypic and molecular traits of Arabidopsis thaliana grown under standardised conditions is still a challenge using climatic devices supplied with common light sources. These are in most cases fluorescent lights, which have several disadvantages such as heat production at higher light intensities, an invariable spectral output, and relatively rapid "ageing". This results in non-desired variations of growth conditions and lowers the comparability of data acquired over extended time periods. In this study, we investigated the growth behaviour of Arabidopsis Col0 under different light conditions, applying fluorescent compared to LED lamps, and we conducted physiological as well as gene expression analyses. By changing the spectral composition and/or light intensity of LEDs we can clearly influence the growth behaviour of Arabidopsis and thereby study phenotypic attributes under very specific light conditions that are stable and reproducible, which is not necessarily given for fluorescent lamps. By using LED lights, we can also roughly mimic the sun light emission spectrum, enabling us to study plant growth in a more natural-like light set-up. We observed distinct growth behaviour under the different light regimes which was reflected by physiological properties of the plants. In conclusion, LEDs provide variable emission spectra for studying plant growth under defined, stable light conditions.

  11. Comparative Phenotypical and Molecular Analyses of Arabidopsis Grown under Fluorescent and LED Light

    Directory of Open Access Journals (Sweden)

    Franka Seiler

    2017-06-01

    Full Text Available Comparative analyses of phenotypic and molecular traits of Arabidopsis thaliana grown under standardised conditions is still a challenge using climatic devices supplied with common light sources. These are in most cases fluorescent lights, which have several disadvantages such as heat production at higher light intensities, an invariable spectral output, and relatively rapid “ageing”. This results in non-desired variations of growth conditions and lowers the comparability of data acquired over extended time periods. In this study, we investigated the growth behaviour of Arabidopsis Col0 under different light conditions, applying fluorescent compared to LED lamps, and we conducted physiological as well as gene expression analyses. By changing the spectral composition and/or light intensity of LEDs we can clearly influence the growth behaviour of Arabidopsis and thereby study phenotypic attributes under very specific light conditions that are stable and reproducible, which is not necessarily given for fluorescent lamps. By using LED lights, we can also roughly mimic the sun light emission spectrum, enabling us to study plant growth in a more natural-like light set-up. We observed distinct growth behaviour under the different light regimes which was reflected by physiological properties of the plants. In conclusion, LEDs provide variable emission spectra for studying plant growth under defined, stable light conditions.

  12. Compensation of optical heterogeneity-induced artifacts in fluorescence molecular tomography: theory and in vivo validation.

    Science.gov (United States)

    Mohajerani, Pouyan; Adibi, Ali; Kempner, Joshua; Yared, Wael

    2009-01-01

    We present a method for reduction of image artifacts induced by the optical heterogeneities of tissue in fluorescence molecular tomography (FMT) through identification and compensation of image regions that evidence propagation of emission light through thin or low-absorption tunnels in tissue. The light tunneled as such contributes to the emission image as spurious components that might substantially overwhelm the desirable fluorescence emanating from the targeted lesions. The proposed method makes use of the strong spatial correlation between the emission and excitation images to estimate the tunneled components and yield a residual image that mainly consists of the signal due to the desirable fluorescence. This residual image is further refined using a coincidence mask constructed for each excitation-emission image pair. The coincidence mask is essentially a map of the "hot spots" that occur in both excitation and emission images, as such areas are often associated with tunneled emission. In vivo studies are performed on a human colon adenocarcinoma xenograft tumor model with subcutaneous tumors and a murine breast adenocarcinoma model with aggressive tumor cell metastasis and growth in the lungs. Results demonstrate significant improvements in the reconstructions achieved by the proposed method.

  13. Encapsulation of serotonin in β-cyclodextrin nano-cavities: Fluorescence spectroscopic and molecular modeling studies

    Science.gov (United States)

    Chaudhuri, Sudip; Chakraborty, Sandipan; Sengupta, Pradeep K.

    2010-06-01

    Serotonin is a physiologically important biogenic amine, deficiency of which leads to mental disorders such as Alzheimer's disease, schizophrenia, infantile autism, and depression. Both β-cyclodextrin (β-CD) and its chemically substituted synthetic varieties (often possessing enhanced aqueous solubility and improved drug complexing abilities) are finding wide applications as drug delivery vehicles. Here we have studied the encapsulation of serotonin in β-CD and succinyl-2-hydroxypropyl β-cyclodextrin (SHP-β-CD) by exploiting the intrinsic serotonin fluorescence. Enhanced fluorescence emission intensity (which increases by ˜18% and 34% in β-CD and SHPβ-CD respectively) and anisotropy ( r) ( r = 0.075 and 0.1 in β-CD and SHPβ-CD respectively) are observed in presence of the cyclodextrins. From the fluorescence data host-guest interaction with 1:1 stoichiometry is evident, the association constants ( K) being 126.06 M -1 and 461.62 M -1 for β-CD and SHPβ-CD respectively. Additionally, molecular docking and semiempirical calculations have been carried out which provide, for the first time, detailed insights regarding the encapsulation process. In particular, it is evident that the indole ring is inserted within the β-CD cavity with the aliphatic amine side chain protruding towards the primary rim of the β-CD cavity. Docking calculations reveal that hydrogen bonding interactions are involved in the formation of the inclusion complex. Semiempirical calculations indicate that formation of the 1:1 inclusion complex is energetically favorable which is consistent with the fluorescence data.

  14. EDTA aggregates induce SYPRO orange-based fluorescence in thermal shift assay.

    Directory of Open Access Journals (Sweden)

    Tobias Kroeger

    Full Text Available Ethylenediaminetetraacetic acid (EDTA is widely used in the life sciences as chelating ligand of metal ions. However, formation of supramolecular EDTA aggregates at pH > 8 has been reported, which may lead to artifactual assay results. When applied as a buffer component at pH ≈ 10 in differential scanning fluorimetry (TSA using SYPRO Orange as fluorescent dye, we observed a sharp change in fluorescence intensity about 20°C lower than expected for the investigated protein. We hypothesized that this change results from SYPRO Orange/EDTA interactions. TSA experiments in the presence of SYPRO Orange using solutions that contain EDTA-Na+ but no protein were performed. The TSA experiments provide evidence that suggests that at pH > 9, EDTA4- interacts with SYPRO Orange in a temperature-dependent manner, leading to a fluorescence signal yielding a "denaturation temperature" of ~68°C. Titrating Ca2+ to SYPRO Orange and EDTA solutions quenched fluorescence. Ethylene glycol tetraacetic acid (EGTA behaved similarly to EDTA. Analytical ultracentrifugation corroborated the formation of EDTA aggregates. Molecular dynamics simulations of free diffusion of EDTA-Na+ and SYPRO Orange of in total 27 μs suggested the first structural model of EDTA aggregates in which U-shaped EDTA4- arrange in an inverse bilayer-like manner, exposing ethylene moieties to the solvent, with which SYPRO Orange interacts. We conclude that EDTA aggregates induce a SYPRO Orange-based fluorescence in TSA. These results make it relevant to ascertain that future TSA results are not influenced by interference between EDTA, or EDTA-related molecules, and the fluorescent dye.

  15. In vivo quantification of fluorescent molecular markers in real-time by ratio Imaging for diagnostic screening and image-guided surgery

    NARCIS (Netherlands)

    Bogaards, A.; Sterenborg, H. J. C. M.; Trachtenberg, J.; Wilson, B. C.; Lilge, L.

    2007-01-01

    Future applications of "molecular diagnostic screening" and "molecular image-guided surgery" will demand images of molecular markers with high resolution and high throughput (similar to >= 30 frames/second). MRI, SPECT, PET, optical fluorescence tomography, hyper-spectral fluorescence imaging, and

  16. Fluorescence based fiber optic and planar waveguide biosensors. A review.

    Science.gov (United States)

    Benito-Peña, Elena; Valdés, Mayra Granda; Glahn-Martínez, Bettina; Moreno-Bondi, Maria C

    2016-11-02

    The application of optical biosensors, specifically those that use optical fibers and planar waveguides, has escalated throughout the years in many fields, including environmental analysis, food safety and clinical diagnosis. Fluorescence is, without doubt, the most popular transducer signal used in these devices because of its higher selectivity and sensitivity, but most of all due to its wide versatility. This paper focuses on the working principles and configurations of fluorescence-based fiber optic and planar waveguide biosensors and will review biological recognition elements, sensing schemes, as well as some major and recent applications, published in the last ten years. The main goal is to provide the reader a general overview of a field that requires the joint collaboration of researchers of many different areas, including chemistry, physics, biology, engineering, and material science. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. ICT-Isomerization-Induced Turn-On Fluorescence Probe with a Large Emission Shift for Mercury Ion: Application in Combinational Molecular Logic.

    Science.gov (United States)

    Bhatta, Sushil Ranjan; Mondal, Bijan; Vijaykumar, Gonela; Thakur, Arunabha

    2017-10-02

    A unique turn-on fluorescent device based on a ferrocene-aminonaphtholate derivative specific for Hg 2+ cation was developed. Upon binding with Hg 2+ ion, the probe shows a dramatic fluorescence enhancement (the fluorescence quantum yield increases 58-fold) along with a large red shift of 68 nm in the emission spectrum. The fluorescence enhancement with a red shift may be ascribed to the combinational effect of C═N isomerization and an extended intramolecular charge transfer (ICT) mechanism. The response was instantaneous with a detection limit of 2.7 × 10 -9 M. Upon Hg 2+ recognition, the ferrocene/ferrocenium redox peak was anodically shifted by ΔE 1/2 = 72 mV along with a "naked eye" color change from faint yellow to pale orange for this metal cation. Further, upon protonation of the imine nitrogen, the present probe displays a high fluorescence output due to suppression of the C═N isomerization process. Upon deprotonation using strong base, the fluorescence steadily decreases, which indicates that H + and OH - can be used to regulate the off-on-off fluorescence switching of the present probe. Density functional theory studies revealed that the addition of acid leads to protonation of the imine N (according to natural bond orbital analysis), and the resulting iminium proton forms a strong H-bond (2.307 Å) with one of the triazole N atoms to form a five-membered ring, which makes the molecule rigid; hence, enhancement of the ICT process takes place, thereby leading to a fluorescence enhancement with a red shift. The unprecedented combination of H + , OH - , and Hg 2+ ions has been used to generate a molecular system exhibiting the INHIBIT-OR combinational logic operation.

  18. Dissolved organic matter characteristics along sabo dammed streams based on ultraviolet visible and fluorescence spectral properties.

    Science.gov (United States)

    Praise, Susan; Ito, Hiroaki; An, Ying; Watanabe, Kazuya; Watanabe, Toru

    2018-02-17

    Changes in dissolved organic matter (DOM) characteristics were investigated in two mountainous streams with closed-type sabo dams. Surface water was collected from four stations along the two mountainous streams and analyzed using ultraviolet-visible spectrophotometry and excitation-emission fluorescence matrix (EEM) methods. Optical properties of DOM indicated an increase in molecular weight and aromaticity at stations near the sabo dams. Average spectral ratio values were low before and after the dam (i.e., Specific ultraviolet absorbance (SUVA 254 ) increased in the vicinities of the dams. While chromophoric DOM characteristics from two sites were influenced by the dam, fluorescence components, however, did not show notable changes around dams. Instead, the three chromophoric components distinguished by EEM-parallel factor analysis, that is, humic-like (C1 and C2) and protein-like (C3) increase along the stream. Fulvic-like component (C1) had a high fluorescence intensity at all stations; all the three components were more abundant in the downstream section. Chromophoric DOM properties varied along the stream based on alterations in molecular size and aromaticity. Using multivariate analysis, the studied sites were grouped into three clusters related to sabo dams and other activities. We conclude that sabo dams modify DOM characteristics which influence the behavior of DOM transported along the stream.

  19. Rational molecular design towards Vis/NIR absorption and fluorescence by using pyrrolopyrrole aza-BODIPY and its highly conjugated structures for organic photovoltaics.

    Science.gov (United States)

    Shimizu, Soji; Iino, Taku; Saeki, Akinori; Seki, Shu; Kobayashi, Nagao

    2015-02-09

    Pyrrolopyrrole aza-BODIPY (PPAB) developed in our recent study from diketopyrrolopyrrole by titanium tetrachloride-mediated Schiff-base formation reaction with heteroaromatic amines is a highly potential chromophore due to its intense absorption and fluorescence in the visible region and high fluorescence quantum yield, which is greater than 0.8. To control the absorption and fluorescence of PPAB, particularly in the near-infrared (NIR) region, further molecular design was performed using DFT calculations. This results in the postulation that the HOMO-LUMO gap of PPAB is perturbed by the heteroaromatic moieties and the aryl-substituents. Based on this molecular design, a series of new PPAB molecules was synthesized, in which the largest redshifts of the absorption and fluorescence maxima up to 803 and 850 nm, respectively, were achieved for a PPAB consisting of benzothiazole rings and terthienyl substituents. In contrast to the sharp absorption of PPAB, a PPAB dimer, which was prepared by a cross-coupling reaction of PPAB monomers, exhibited panchromatic absorption across the UV/Vis/NIR regions. With this series of PPAB chromophores in hand, a potential application of PPAB as an optoelectronic material was investigated. After identifying a suitable PPAB molecule for application in organic photovoltaic cells based on evaluation using time-resolved microwave conductivity measurements, a maximized power conversion efficiency of 1.27 % was achieved. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Fluorescence based fiber optic and planar waveguide biosensors. A review

    Energy Technology Data Exchange (ETDEWEB)

    Benito-Peña, Elena [Department of Analytical Chemistry, Faculty of Chemistry, Complutense University, 28040 Madrid (Spain); Valdés, Mayra Granda [Department of Analytical Chemistry, Faculty of Chemistry, University of La Habana, 10400 La Habana (Cuba); Glahn-Martínez, Bettina [Department of Analytical Chemistry, Faculty of Chemistry, Complutense University, 28040 Madrid (Spain); Moreno-Bondi, Maria C., E-mail: mcmbondi@quim.ucm.es [Department of Analytical Chemistry, Faculty of Chemistry, Complutense University, 28040 Madrid (Spain)

    2016-11-02

    The application of optical biosensors, specifically those that use optical fibers and planar waveguides, has escalated throughout the years in many fields, including environmental analysis, food safety and clinical diagnosis. Fluorescence is, without doubt, the most popular transducer signal used in these devices because of its higher selectivity and sensitivity, but most of all due to its wide versatility. This paper focuses on the working principles and configurations of fluorescence-based fiber optic and planar waveguide biosensors and will review biological recognition elements, sensing schemes, as well as some major and recent applications, published in the last ten years. The main goal is to provide the reader a general overview of a field that requires the joint collaboration of researchers of many different areas, including chemistry, physics, biology, engineering, and material science. - Highlights: • Principles, configurations and fluorescence techniques using fiber optic and planar waveguide biosensors are discussed. • The biorecognition elements and sensing schemes used in fiber optic and planar waveguide platforms are reviewed. • Some major and recent applications of fiber optic and planar waveguide biosensors are introduced.

  1. Photolysis study of octyl p-methoxycinnamate loaded microemulsion by molecular fluorescence and chemometric approach.

    Science.gov (United States)

    Nascimento, Danielle Silva; Insausti, Matías; Band, Beatriz Susana Fernández; Grünhut, Marcos

    2018-02-15

    Octyl p-methoxycinnamate (OMC) is one of the most widely used sunscreen agents. However, the efficiency of OMC as UV filter over time is affected due to the formation of the cis-isomer which presents a markedly lower extinction coefficient (ε cis =12,600L mol -1 cm -1 at 291nm) than the original trans-isomer (ε trans =24,000L mol -1 cm -1 at 310nm). In this work, a novel carrier for OMC based on an oil-in-water microemulsion is proposed in order to improve the photostability of this sunscreen. The formulation was composed of 29.2% (w/w) of a 3:1 mixture of ethanol (co-surfactant) and decaethylene glycol mono-dodecyl ether (surfactant), 1.5% (w/w) of oleic acid (oil phase) and 69.2% (w/w) of water. This microemulsion was prepared in a simple way, under moderate stirring at 25°C and using acceptable, biocompatible and accessible materials for topical use. OMC was incorporated in the vehicle at a final concentration of 5.0% (w/w), taking into account the maximum permitted levels established by international norms. Then, a photolysis study of the loaded formulation was performed using a continuous flow system. The direct photolysis was monitored over time by molecular fluorescence. The recorded spectra data between 370 y 490nm were analyzed by multivariate curve resolution-alternating least squares algorithm. The kinetic rate constants corresponding to the photolysis of the trans-OMC were calculated from the concentration profiles, resulting in 0.0049s -1 for the trans-OMC loaded microemulsion and 0.0131s -1 for the trans-OMC in aqueous media. These results demonstrate a higher photostability of the trans-OMC when loaded in the proposed vehicle with respect to the free trans-OMC in aqueous media. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. An off-on fluorescence probe targeting mitochondria based on oxidation-reduction response for tumor cell and tissue imaging

    Science.gov (United States)

    Yao, Hanchun; Cao, Li; Zhao, Weiwei; Zhang, Suge; Zeng, Man; Du, Bin

    2017-10-01

    In this study, a tumor-targeting poly( d, l-lactic-co-glycolic acid) (PLGA) loaded "off-on" fluorescent probe nanoparticle (PFN) delivery system was developed to evaluate the region of tumor by off-on fluorescence. The biodegradability of the nanosize PFN delivery system readily released the probe under tumor acidic conditions. The probe with good biocompatibility was used to monitor the intracellular glutathione (GSH) of cancer cells and selectively localize to mitochondria for tumor imaging. The incorporated tumor-targeting probe was based on the molecular photoinduced electron transfer (PET) mechanism preventing fluorescence ("off" state) and could be easily released under tumor acidic conditions. However, the released tumor-targeting fluorescence probe molecule was selective towards GSH with high selectivity and an ultra-sensitivity for the mitochondria of cancer cells and tissues significantly increasing the probe molecule fluorescence signal ("on" state). The tumor-targeting fluorescence probe showed sensitivity to GSH avoiding interference from cysteine and homocysteine. The PFNs could enable fluorescence-guided cancer imaging during cancer therapy. This work may expand the biological applications of PFNs as a diagnostic reagent, which will be beneficial for fundamental research in tumor imaging. [Figure not available: see fulltext.

  3. Fluorescent bioaerosol particle, molecular tracer, and fungal spore concentrations during dry and rainy periods in a semi-arid forest

    Directory of Open Access Journals (Sweden)

    M. I. Gosselin

    2016-12-01

    Full Text Available Bioaerosols pose risks to human health and agriculture and may influence the evolution of mixed-phase clouds and the hydrological cycle on local and regional scales. The availability and reliability of methods and data on the abundance and properties of atmospheric bioaerosols, however, are rather limited. Here we analyze and compare data from different real-time ultraviolet laser/light-induced fluorescence (UV-LIF instruments with results from a culture-based spore sampler and offline molecular tracers for airborne fungal spores in a semi-arid forest in the southern Rocky Mountains of Colorado. Commercial UV-APS (ultraviolet aerodynamic particle sizer and WIBS-3 (wideband integrated bioaerosol sensor, version 3 instruments with different excitation and emission wavelengths were utilized to measure fluorescent aerosol particles (FAPs during both dry weather conditions and periods heavily influenced by rain. Seven molecular tracers of bioaerosols were quantified by analysis of total suspended particle (TSP high-volume filter samples using a high-performance anion-exchange chromatography system with pulsed amperometric detection (HPAEC-PAD. From the same measurement campaign, Huffman et al. (2013 previously reported dramatic increases in total and fluorescent particle concentrations during and immediately after rainfall and also showed a strong relationship between the concentrations of FAPs and ice nuclei (Huffman et al., 2013; Prenni et al., 2013. Here we investigate molecular tracers and show that during rainy periods the atmospheric concentrations of arabitol (35.2 ± 10.5 ng m−3 and mannitol (44.9 ± 13.8 ng m−3 were 3–4 times higher than during dry periods. During and after rain, the correlations between FAP and tracer mass concentrations were also significantly improved. Fungal spore number concentrations on the order of 104 m−3, accounting for 2–5 % of TSP mass during dry periods and 17–23 % during rainy

  4. A Far-ultraviolet Fluorescent Molecular Hydrogen Emission Map of the Milky Way Galaxy

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Young-Soo; Min, Kyoung-Wook [Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701 (Korea, Republic of); Seon, Kwang-Il; Han, Wonyong [Korea Astronomy and Space Science Institute (KASI), 776 Daedeokdae-ro, Yuseong-gu, Daejeon, 305-348 (Korea, Republic of); Edelstein, Jerry, E-mail: stspeak@gmail.com [Space Sciences Laboratory, University of California, Berkeley, CA, 94720 (United States)

    2017-08-01

    We present the far-ultraviolet (FUV) fluorescent molecular hydrogen (H{sub 2}) emission map of the Milky Way Galaxy obtained with FIMS/SPEAR covering ∼76% of the sky. The extinction-corrected intensity of the fluorescent H{sub 2} emission has a strong linear correlation with the well-known tracers of the cold interstellar medium (ISM), including color excess E(B–V) , neutral hydrogen column density N (H i), and H α emission. The all-sky H{sub 2} column density map was also obtained using a simple photodissociation region model and interstellar radiation fields derived from UV star catalogs. We estimated the fraction of H{sub 2} ( f {sub H2}) and the gas-to-dust ratio (GDR) of the diffuse ISM. The f {sub H2} gradually increases from <1% at optically thin regions where E(B–V) < 0.1 to ∼50% for E(B–V)  = 3. The estimated GDR is ∼5.1 × 10{sup 21} atoms cm{sup −2} mag{sup −1}, in agreement with the standard value of 5.8 × 10{sup 21} atoms cm{sup −2} mag{sup −1}.

  5. Fluorescent Molecular Rotor-in-Paraffin Waxes for Thermometry and Biometric Identification.

    Science.gov (United States)

    Jin, Young-Jae; Dogra, Rubal; Cheong, In Woo; Kwak, Giseop

    2015-07-08

    Novel thermoresponsive sensor systems consisting of a molecular rotor (MR) and paraffin wax (PW) were developed for various thermometric and biometric identification applications. Polydiphenylacetylenes (PDPAs) coupled with long alkyl chains were used as MRs, and PWs of hydrocarbons having 16-20 carbons were utilized as phase-change materials. The PDPAs were successfully dissolved in the molten PWs and did not act as an impurity that prevents phase transition of the PWs. These PDPA-in-PW hybrids had almost the same enthalpies and phase-transition temperatures as the corresponding pure PWs. The hybrids exhibited highly reversible fluorescence (FL) changes at the critical temperatures during phase transition of the PWs. These hybrids were impregnated into common filter paper in the molten state by absorption or were encapsulated into urea resin to enhance their mechanical integrity and cyclic stability during repeated use. The wax papers could be utilized in highly advanced applications including FL image writing/erasing, an array-type thermo-indicator, and fingerprint/palmprint identification. The present findings should facilitate the development of novel fluorescent sensor systems for biometric identification and are potentially applicable for biological and biomedical thermometry.

  6. Expression-Enhanced Fluorescent Proteins Based on Enhanced Green Fluorescent Protein for Super-resolution Microscopy.

    Science.gov (United States)

    Duwé, Sam; De Zitter, Elke; Gielen, Vincent; Moeyaert, Benjamien; Vandenberg, Wim; Grotjohann, Tim; Clays, Koen; Jakobs, Stefan; Van Meervelt, Luc; Dedecker, Peter

    2015-10-27

    "Smart fluorophores", such as reversibly switchable fluorescent proteins, are crucial for advanced fluorescence imaging. However, only a limited number of such labels is available, and many display reduced biological performance compared to more classical variants. We present the development of robustly photoswitchable variants of enhanced green fluorescent protein (EGFP), named rsGreens, that display up to 30-fold higher fluorescence in E. coli colonies grown at 37 °C and more than 4-fold higher fluorescence when expressed in HEK293T cells compared to their ancestor protein rsEGFP. This enhancement is not due to an intrinsic increase in the fluorescence brightness of the probes, but rather due to enhanced expression levels that allow many more probe molecules to be functional at any given time. We developed rsGreens displaying a range of photoswitching kinetics and show how these can be used for multimodal diffraction-unlimited fluorescence imaging such as pcSOFI and RESOLFT, achieving a spatial resolution of ∼70 nm. By determining the first ever crystal structures of a negative reversibly switchable FP derived from Aequorea victoria in both the "on"- and "off"-conformation we were able to confirm the presence of a cis-trans isomerization and provide further insights into the mechanisms underlying the photochromism. Our work demonstrates that genetically encoded "smart fluorophores" can be readily optimized for biological performance and provides a practical strategy for developing maturation- and stability-enhanced photochromic fluorescent proteins.

  7. Sensitive turn-on fluorescent detection of tartrazine based on fluorescence resonance energy transfer.

    Science.gov (United States)

    Huang, Sheng Tian; Shi, Yan; Li, Nian Bing; Luo, Hong Qun

    2012-01-18

    We introduce a sensitive, rapid, label-free and general fluorescent method for the determination of tartrazine by competitive binding to reduced graphene oxide (rGO) against fluorescein, and the fluorescence recovery upon fluorescein desorption from rGO provides a quantitative readout for tartrazine, giving a detection limit of 0.53 ng mL(-1).

  8. A New Anthracene Based Fluorescent Turn-On Sensor for Fe{sup 3+}

    Energy Technology Data Exchange (ETDEWEB)

    An, Kyounglyong; Lee, Sangoh; Koh, Sun Duk; Shin, Seungrim; Shin, Jongil; Park, Sooyoul; Jun, Kun [Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of); Son, Younga; Park, Koon Ha [Chungnam National Univ., Daejeon (Korea, Republic of)

    2014-01-15

    We have developed a new fluorescent sensor for Fe3+ based on anthracene moiety of hydroxyalkylamine. Fluorescent sensor 1 displayed highly selective and sensitive fluorescent 'turn-on' toward Fe3+ in acetonitrile solution. The 1:1 stoichiometry of sensor complex 1+Fe3+ was confirmed by Job's plots based on fluorescence emission titration. Furthermore, the binding sites of sensor complex 1+Fe3+ was properly identified from 1H NMR study.

  9. Fluorescent signaling based on sulfoxide profluorophores: application to the visual detection of the explosive TATP.

    Science.gov (United States)

    Malashikhin, Sergey; Finney, Nathaniel S

    2008-10-01

    The first visual fluorescence-based assay for the peroxide explosive triacetone triperoxide (TATP) is described. The assay is based on a conceptually new fluorescence signaling mechanism, in which nonemissive pyrenyl sulfoxide profluorophores are oxidized to visibly emissive pyrenyl sulfones. Although not without limitations, these first-generation fluorescent probes can provide a visual response to ca. 100 nmol of TATP. In addition, the success of this assay suggests the potential for broader application of aryl sulfoxides in fluorescent chemosensing.

  10. Development of an oxygen-independent flavin mononucleotide-based fluorescent reporter system in Clostridium beijerinckii and its potential applications.

    Science.gov (United States)

    Seo, Seung-Oh; Lu, Ting; Jin, Yong-Su; Blaschek, Hans P

    2018-01-10

    Clostridium beijerinckii is a predominant solventogenic clostridia with great attraction for renewable liquid biofuel and biochemical production. Metabolic engineering and synthetic biology can be employed to engineer the strain toward desirable phenotypes. However, current limited information such as promoter strength and gene regulation may hinder the efficient engineering of the strain. To investigate genetic information and complex cellular bioprocesses of C. beijerinckii, an in vivo fluorescence reporter system can be employed. In general, green fluorescence protein (GFP) and relative analogs have been widely used as real-time reporters. However, GFP-family proteins require molecular oxygen for fluorescence maturation. Considering the strict anaerobic growth requirement of the clostridia, an oxygen-independent fluorescence reporter such as a flavin mononucleotide-based fluorescent protein (FbFP) can be used as an alternative fluorescence reporter. In this study, we synthesized and expressed the codon-optimized FbFP gene for C. beijerinckii (CbFbFP) based on the nucleotide sequence of Bacillus subtilis YtvA variant EcFbFP in C. beijerinckii NCIMB 8052 wild-type. Protein expression and in vivo fluorescence of CbFbFP in C. beijeirnckii were confirmed under anaerobic growth conditions. Through fluorescence-activated cell sorting (FACS), we isolated the bright cells from the heterogenous population of C. beijerinckii cells expressing CbFbFP. Several mutations were found in the isolated plasmid which may be responsible for the high-level expression of CbFbFP in C. beijerinckii. The mutant plasmid and CbFbFP reporter were further utilized for strain selection, real-time fluorescence measurement, population analysis, and metabolic engineering in this study. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Hybrid Integrated Silicon Microfluidic Platform for Fluorescence Based Biodetection

    Directory of Open Access Journals (Sweden)

    André Darveau

    2007-09-01

    Full Text Available The desideratum to develop a fully integrated Lab-on-a-chip device capable ofrapid specimen detection for high throughput in-situ biomedical diagnoses and Point-of-Care testing applications has called for the integration of some of the novel technologiessuch as the microfluidics, microphotonics, immunoproteomics and Micro ElectroMechanical Systems (MEMS. In the present work, a silicon based microfluidic device hasbeen developed for carrying out fluorescence based immunoassay. By hybrid attachment ofthe microfluidic device with a Spectrometer-on-chip, the feasibility of synthesizing anintegrated Lab-on-a-chip type device for fluorescence based biosensing has beendemonstrated. Biodetection using the microfluidic device has been carried out usingantigen sheep IgG and Alexafluor-647 tagged antibody particles and the experimentalresults prove that silicon is a compatible material for the present application given thevarious advantages it offers such as cost-effectiveness, ease of bulk microfabrication,superior surface affinity to biomolecules, ease of disposability of the device etc., and is thussuitable for fabricating Lab-on-a-chip type devices.

  12. A small azide-modified thiazole-based reporter molecule for fluorescence and mass spectrometric detection

    Directory of Open Access Journals (Sweden)

    Stefanie Wolfram

    2014-10-01

    Full Text Available Molecular probes are widely used tools in chemical biology that allow tracing of bioactive metabolites and selective labeling of proteins and other biomacromolecules. A common structural motif for such probes consists of a reporter that can be attached by copper(I-catalyzed 1,2,3-triazole formation between terminal alkynes and azides to a reactive headgroup. Here we introduce the synthesis and application of the new thiazole-based, azide-tagged reporter 4-(3-azidopropoxy-5-(4-bromophenyl-2-(pyridin-2-ylthiazole for fluorescence, UV and mass spectrometry (MS detection. This small fluorescent reporter bears a bromine functionalization facilitating the automated data mining of electrospray ionization MS runs by monitoring for its characteristic isotope signature. We demonstrate the universal utility of the reporter for the detection of an alkyne-modified small molecule by LC–MS and for the visualization of a model protein by in-gel fluorescence. The novel probe advantageously compares with commercially available azide-modified fluorophores and a brominated one. The ease of synthesis, small size, stability, and the universal detection possibilities make it an ideal reporter for activity-based protein profiling and functional metabolic profiling.

  13. A Cu2+-selective fluorescent chemosensor based on BODIPY with two pyridine ligands and logic gate

    Science.gov (United States)

    Huang, Liuqian; Zhang, Jing; Yu, Xiaoxiu; Ma, Yifan; Huang, Tianjiao; Shen, Xi; Qiu, Huayu; He, Xingxing; Yin, Shouchun

    2015-06-01

    A novel near-infrared fluorescent chemosensor based on BODIPY (Py-1) has been synthesized and characterized. Py-1 displays high selectivity and sensitivity for sensing Cu2+ over other metal ions in acetonitrile. Upon addition of Cu2+ ions, the maximum absorption band of Py-1 in CH3CN displays a red shift from 603 to 608 nm, which results in a visual color change from pink to blue. When Py-1 is excited at 600 nm in the presence of Cu2+, the fluorescent emission intensity of Py-1 at 617 nm is quenched over 86%. Notably, the complex of Py-1-Cu2+ can be restored with the introduction of EDTA or S2-. Consequently, an IMPLICATION logic gate at molecular level operating in fluorescence mode with Cu2+ and S2- as chemical inputs can be constructed. Finally, based on the reversible and reproducible system, a nanoscale sequential memory unit displaying "Writing-Reading-Erasing-Reading" functions can be integrated.

  14. [Molecular bases of prion diseases].

    Science.gov (United States)

    Pokrovskiĭ, V I; Kiselev, O I

    1998-01-01

    The paper briefly analyzes the origin of priones and their association with the cellular gene and homologous protein of diseases in man and animals. There is evidence for a direct relationship of the agents that cause spongious encephalitis in the cattle and a new type of Creutzfeldt-Jacob disease in man. The molecular organization of priones and the conformational cellular protein changes underlying the infectious activation of the cell homologue of priones. Emphasis is first laid on the capacity of the cell homologue of priones and their infectiously active derivative to bind to DNA or RNA. In the context of concepts of the priones yeasts an attempt was made to explain the reproduction through the altered control of translation of mRNA that encodes the cellular homologue of priones, which accounts for the duration of the incubation period of the disease. The infections caused by priones are referred to as the so-called slow infections. But in the context of the proposed hypothesis, an infective process in the tissues did not really have some typical signs of infection and resembles accumulation diseases more without the replicative burst typical of infectious processes. The paper gives data on the vital cycle of priones in infected animals and changes in the accumulation of an infective agent. This assesses the currently available diagnostic methods and gives preference to the methods which will be based on the use of monoclonal antibodies that specifically recognize the conformationally altered form of an infectious prione or on the identification of primary oligomeric forms which manifest the onset of amyloidization of the damaged tissues. The main conclusion of the paper is that protein prionization is a common biological phenomenon and the diseases caused by these processes will increase in number in the near future, which makes it necessary to develop diagnostic methods and universal treatments of diseases, such as bacterial infections by using antibiotics.

  15. Bases moleculares de las leucemias agudas

    Directory of Open Access Journals (Sweden)

    G. Martínez Antuña

    2006-04-01

    Full Text Available El gran desarrollo de la biología molecular en los últimos años ha contribuido a un importante avance en los conocimientos relacionados con las bases moleculares de las leucemias agudas (LA. Ademas de profundizar en la biología de estas enfermedades y conocer las bases moleculares, ha renido también gran impacto en mejorar el resultado de los tratamientos y disminuir la toxicidad de las terapias.

  16. [Fluorescence lifetime measurement based on different pump waveform].

    Science.gov (United States)

    Huang, Xiu-jun; Chen, Jian-guo; Feng, Guo-ying; Yang, Li-ling; Deng, Guo-liang; Tang, Xiao-jun; Zhou, Shou-huan

    2010-11-01

    The present paper analyzes the different pump wave theory under the fluorescence decay law, and presents a different pump wave measured fluorescence lifetime of the new method-double-pulse detection method, namely the use of pump pulse and probe pulse for the fluorescence decay methods to measure fluorescence lifetime. By measuring the sample neodymium glass and Cr:ZnSe crystal fluorescence lifetime, results showed that: using this method can be achieved under different pump wave in visible and near-infrared to the mid-infrared laser medium of the fluorescence lifetime measurement. Therefore, using the measurement method can be convenient and effective to avoid seeking samples by deconvolution for fluorescence lifetime with the cumbersome process of measuring laser medium at different pump wave under the fluorescence lifetime with a reference value.

  17. First molecular identification of the transgene red fluorescent protein (RFP in transgenic ornamental zebrafish (Danio rerio introduced in Peru

    Directory of Open Access Journals (Sweden)

    Carlos Scotto

    2013-09-01

    Full Text Available In this paper the transgenic fluorescent red, orange and pink zebra fish (Danio rerio, found in local aquariums in Peru, were identified using the PCR technique to amplify the transgene RFP sea anemone belonging to Discosoma spp. The gene expression of the red fluorescent protein (RFP transgene was found to determine different gradients-of-bioluminescence (shades in color in each GMO fish analyzed. We performed sequence analysis of the two variants of the RFP along with six variants of the existing fluorescent protein GFP from the Genbank, this could help identify quickly if they are new genes or variants thereof as these novel fluorescent proteins may be introduced in aquatic GMO in the future. Thus, developing and improving biosecurity measures through its timely detection at the molecular genetic level.

  18. Silica nanodisks as platforms for fluorescence lifetime-based ...

    Indian Academy of Sciences (India)

    These dyes exhibit pH-dependent fluorescence properties. The nanoconjugates are found to sense the pH of the medium, through systematic variation of the fluorescence intensity ratios of the reporter dye at the surface and the reference dye at the core. Moreover, the fluorescence lifetimes and corresponding amplitudes of ...

  19. Biochemical activity of a fluorescent dye rhodamine 6G: Molecular modeling, electrochemical, spectroscopic and thermodynamic studies.

    Science.gov (United States)

    Al Masum, Abdulla; Chakraborty, Maharudra; Ghosh, Soumen; Laha, Dipranjan; Karmakar, Parimal; Islam, Md Maidul; Mukhopadhyay, Subrata

    2016-11-01

    Interaction of CT DNA with Rhodamine 6G (R6G) has been studied using molecular docking, electrochemical, spectroscopic and thermodynamic methods. From the study, it was illustrated that Rhodamine 6G binds to the minor groove of CT DNA. The binding was cooperative in nature. Circular voltametric study showed significant change in peak current and peak potential due to complexation. All the studies showed that the binding constant was in the order of 10 6 M -1 . Circular dichroic spectra showed significant conformational change on binding and DNA unwind during binding. Thermodynamic study showed that binding was favored by negative enthalpy and positive entropy change. From thermodynamic study it was also observed that several positive and negative free energies played significant role during binding and the unfavorable conformational free energy change was overcame by highly negative hydrophobic and salt dependent free energy changes. The experimental results were further validated using molecular docking study and the effect of structure on binding has been studied theoretically. From docking study it was found that the hydrophobic interaction and hydrogen bonds played a significant role during binding. The dye was absorbed by cell and this phenomenon was studied using fluorescent microscope. Cell survivability test showed that the dye active against Human Breast Cancer cells MDA-MB 468. ROS study showed that the activity is due to the production of reactive oxygen. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Value of comparative genomic hybridization and fluorescence in situ hybridization for molecular diagnostics in multiple myeloma.

    Science.gov (United States)

    Liebisch, Peter; Viardot, Andreas; Bassermann, Nicole; Wendl, Christiane; Roth, Katrin; Goldschmidt, Hartmut; Einsele, Hermann; Straka, Christian; Stilgenbauer, Stephan; Döhner, Hartmut; Bentz, Martin

    2003-07-01

    Chromosomal abnormalities, such as 13q deletions, are emerging as important prognostic factors in multiple myeloma. Fluorescence in situ hybridization (FISH) using specific DNA probes is the technique most widely used for the determination of genomic aberrations in this disease. The utility of comparative genomic hybridization (CGH) for molecular diagnostics in plasma cell malignancies has not been systematically analysed. We investigated tumour samples of patients with multiple myeloma (n = 43) or plasma cell leukaemia (n = 3) using CGH and FISH with five DNA probes localized to chromosome bands 1p22, 6q21, 11q22-q23, 13q14 and 17p13. By CGH, the most frequent genomic changes were gains on chromosomes 1q, 9q and 11q, as well as losses on chromosomes 13q, 6q, Xp and Xq. By FISH, trisomy 11q was identified at a similar frequency to the 13q deletion (42%). Compared with FISH data, the sensitivity of CGH was 80.7% and the specificity was 97.5%. Thirty-two aberrations found by FISH were not identified by CGH, mostly as a result of the proportion of cells carrying the respective aberrations, or because of the limited spatial resolution of CGH. Our data indicate that, for clinical molecular diagnostics in multiple myeloma, FISH with a disease-specific DNA probe set is superior to CGH analysis.

  1. AURAMINE O AS POTENTIAL AMYLOID MARKER: FLUORESCENCE AND MOLECULAR DOCKING STUDIES

    Directory of Open Access Journals (Sweden)

    K. Vus

    2017-10-01

    Full Text Available The applicability of Auramine O to the detection and characterization of lysozyme and serum albumin amyloid fibrils has been assessed using the fluorimetric titration and molecular docking. The parameters of the dye binding to native and fibrillar proteins were estimated in terms of the Langmuir adsorption model. It was found that Auramine O displays the high affinity for amyloid fibrils, being of the same order of magnitude as that of the classical amyloid markers. The dye also showed greater fluorescence response to lysozyme fibrils and lower sensitivity to the native protein, than Thioflavin T. Furthermore, unlike Thioflavin T, Auramine O was able to detect the morphological differences between lysozyme and albumin fibrils due to the shifts in the position of the emission maxima of the fibril-incorporated fluorophore. The molecular docking studies revealed that Auramine O and Thioflavin T form the most stable complexes with the G54_L56/S60_W62 groove of lysozyme fibrils, running parallel to the fibril axis. The results obtained suggest the contribution of both hydrophobic and electrostatic interactions to the stabilization of the dye complexes with amyloid fibrils.

  2. Noninvasive imaging of multiple myeloma using near infrared fluorescent molecular probe

    Science.gov (United States)

    Hathi, Deep; Zhou, Haiying; Bollerman-Nowlis, Alex; Shokeen, Monica; Akers, Walter J.

    2016-03-01

    Multiple myeloma is a plasma cell malignancy characterized by monoclonal gammopathy and osteolytic bone lesions. Multiple myeloma is most commonly diagnosed in late disease stages, presenting with pathologic fracture. Early diagnosis and monitoring of disease status may improve quality of life and long-term survival for multiple myeloma patients from what is now a devastating and fatal disease. We have developed a near-infrared targeted fluorescent molecular probe with high affinity to the α4β1 integrin receptor (VLA-4)overexpressed by a majority of multiple myeloma cells as a non-radioactive analog to PET/CT tracer currently being developed for human diagnostics. A near-infrared dye that emits about 700 nm was conjugated to a high affinity peptidomimmetic. Binding affinity and specificity for multiple myeloma cells was investigated in vitro by tissue staining and flow cytometry. After demonstration of sensitivity and specificity, preclinical optical imaging studies were performed to evaluate tumor specificity in murine subcutaneous and metastatic multiple myeloma models. The VLA-4-targeted molecular probe showed high affinity for subcutaneous MM tumor xenografts. Importantly, tumor cells specific accumulation in the bone marrow of metastatic multiple myeloma correlated with GFP signal from transfected cells. Ex vivo flow cytometry of tumor tissue and bone marrow further corroborated in vivo imaging data, demonstrating the specificity of the novel agent and potential for quantitative imaging of multiple myeloma burden in these models.

  3. Aggregation of Oligoarginines at Phospholipid Membranes: Molecular Dynamics Simulations, Time-Dependent Fluorescence Shift, and Biomimetic Colorimetric Assays

    Czech Academy of Sciences Publication Activity Database

    Vazdar, Mario; Wernersson, Erik; Khabiri, Morteza; Cwiklik, Lukasz; Jurkiewicz, Piotr; Hof, Martin; Mann, E.; Kolusheva, S.; Jelinek, R.; Jungwirth, Pavel

    2013-01-01

    Roč. 117, č. 39 (2013), s. 11530-11540 ISSN 1520-6106 R&D Projects: GA MŠk LH12001; GA ČR GBP208/12/G016 Institutional support: RVO:61388963 ; RVO:61388955 Keywords : cell penetrating peptides * oligoarginine * molecular dynamics * membranes * fluorescence spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.377, year: 2013

  4. Development and cytotoxicity of Schiff base derivative as a fluorescence probe for the detection of L-Arginine

    Science.gov (United States)

    Shang, Xuefang; Li, Jie; Guo, Kerong; Ti, Tongyu; Wang, Tianyun; Zhang, Jinlian

    2017-04-01

    Inspired from biological counter parts, chemical modification of Schiff base derivatives with function groups may provide a highly efficient method to detect amino acids. Therefore, a fluorescent probe involving Schiff base and hydroxyl group has been designed and prepared, which showed high response and specificity for Arginine (Arg) among normal eighteen standard kinds of amino acids (Alanine, Valine, Leucine, Isoleucine, Methionine, Asparticacid, Glutamicacid, Arginine, Glycine, Serine, Threonine, Asparagine, Phenylalanine, Histidine, Tryptophan, Proline, Lysine, Glutamine, Tyrosine and Cysteine). Furthermore, theoretical investigation further illustrated the possible binding mode in the host-guest interaction and the roles of molecular frontier orbitals in molecular interplay. In addition, the synthesized fluorescent probe exhibited high binding ability for Arg and low cytotoxicity to MCF-7 cells over a concentration range of 0-200 μg mL-1 which can be also used as a biosensor for the Arg detection in vivo.

  5. Polymerase chain reaction (PCR) based molecular characterization ...

    African Journals Online (AJOL)

    Polymerase chain reaction (PCR) based molecular characterization of popular wheat varieties of Khyber Pukhtunkhwa (KPK) region of Pakistan. ... Molecular markers used in this study show high rate of genetic diversity that can be used to assist a breeding program for the improvement of wheat in KPK-Pakistan. Key words: ...

  6. A critical study on the interactions of hesperitin with human hemoglobin: Fluorescence spectroscopic and molecular modeling approach

    Energy Technology Data Exchange (ETDEWEB)

    Chakraborty, Sandipan [Saroj Mohan Institute of Technology, Hooghly (India); Chaudhuri, Sudip; Pahari, Biswapathik [Biophysics Division, Saha Institute of Nuclear Physics, Kolkata 700064 (India); Taylor, Jasmine [Chemistry Department, Tougaloo College, Tougaloo, MS 39174 (United States); Sengupta, Pradeep K. [Biophysics Division, Saha Institute of Nuclear Physics, Kolkata 700064 (India); Sengupta, Bidisha, E-mail: bsengupta@tougaloo.edu [Chemistry Department, Tougaloo College, Tougaloo, MS 39174 (United States)

    2012-06-15

    Hesperitin, a ubiquitous bioactive flavonoid abundant in citrus fruits is known to possess antioxidant, anti-carcinogenic, hypolipidemic, vasoprotective and other important therapeutic properties. Here we have explored the interactions of hesperitin with normal human hemoglobin (HbA), using steady state and time resolved fluorescence spectroscopy, far UV circular dicroism (CD) spectroscopy, combined with molecular modeling computations. Specific interaction of the flavonoid with HbA is confirmed from flavonoid-induced static quenching which is evident from steady state fluorescence as well as lifetime data. Both temperature dependent fluorescence measurements and molecular docking studies reveal that apart from hydrogen bonding and van der Waals interactions, electrostatic interactions also play crucial role in hesperitin-HbA interactions. Furthermore, electrostatic surface potential calculations indicate that the hesperitin binding site in HbA is intensely positive due to the presence of several lysine and histidine residues. - Highlights: Black-Right-Pointing-Pointer Absorption spectra of hesperitin bound HbA indicates ground state complex formation. Black-Right-Pointing-Pointer Binding induces static quenching of intrinsic fluorescence of the tryptophan of HbA. Black-Right-Pointing-Pointer Molecular docking and electrostatic surface potential calculations were performed. Black-Right-Pointing-Pointer Contrasting binding modes of hesperitin compared to other flavonoids were observed.

  7. A critical study on the interactions of hesperitin with human hemoglobin: Fluorescence spectroscopic and molecular modeling approach

    International Nuclear Information System (INIS)

    Chakraborty, Sandipan; Chaudhuri, Sudip; Pahari, Biswapathik; Taylor, Jasmine; Sengupta, Pradeep K.; Sengupta, Bidisha

    2012-01-01

    Hesperitin, a ubiquitous bioactive flavonoid abundant in citrus fruits is known to possess antioxidant, anti-carcinogenic, hypolipidemic, vasoprotective and other important therapeutic properties. Here we have explored the interactions of hesperitin with normal human hemoglobin (HbA), using steady state and time resolved fluorescence spectroscopy, far UV circular dicroism (CD) spectroscopy, combined with molecular modeling computations. Specific interaction of the flavonoid with HbA is confirmed from flavonoid-induced static quenching which is evident from steady state fluorescence as well as lifetime data. Both temperature dependent fluorescence measurements and molecular docking studies reveal that apart from hydrogen bonding and van der Waals interactions, electrostatic interactions also play crucial role in hesperitin-HbA interactions. Furthermore, electrostatic surface potential calculations indicate that the hesperitin binding site in HbA is intensely positive due to the presence of several lysine and histidine residues. - Highlights: ► Absorption spectra of hesperitin bound HbA indicates ground state complex formation. ► Binding induces static quenching of intrinsic fluorescence of the tryptophan of HbA. ► Molecular docking and electrostatic surface potential calculations were performed. ► Contrasting binding modes of hesperitin compared to other flavonoids were observed.

  8. ESIPT-Based Photoactivatable Fluorescent Probe for Ratiometric Spatiotemporal Bioimaging

    Directory of Open Access Journals (Sweden)

    Xiaohong Zhou

    2016-10-01

    Full Text Available Photoactivatable fluorophores have become an important technique for the high spatiotemporal resolution of biological imaging. Here, we developed a novel photoactivatable probe (PHBT, which is based on 2-(2-hydroxyphenylbenzothiazole (HBT, a small organic fluorophore known for its classic luminescence mechanism through excited-state intramolecular proton transfer (ESIPT with the keto form and the enol form. After photocleavage, PHBT released a ratiometric fluorophore HBT, which showed dual emission bands with more than 73-fold fluorescence enhancement at 512 nm in buffer and more than 69-fold enhancement at 452 nm in bovine serum. The probe displayed a high ratiometric imaging resolution and is believed to have a wide application in biological imaging.

  9. Fluorescence based real time monitoring of fouling in process chromatography

    Science.gov (United States)

    Pathak, Mili; Lintern, Katherine; Chopda, Viki; Bracewell, Daniel G.; Rathore, Anurag S.

    2017-01-01

    A real time monitoring of fouling in liquid chromatography has been presented. The versatility of the approach has been proven by successful implementation in three case studies with an error protein A ligand density and foulant concentration for assessing performance of protein A chromatography resin during purification of monoclonal antibodies. The observations have been supported from LC-MS/MS studies that were independently performed. The second application involves monitoring of foulant deposition during multimode cation exchange chromatography based purification of human serum albumin. Finally, in the third application, monitoring of foulants during multimodal hydrophobic interaction chromatography of recombinant human granulocyte colony stimulating factor is demonstrated. In all three cases, it is observed that the fluorescence intensity consistently increases with resin reuse as more foulants are deposited over time. The proposed approach can be readily used for real time monitoring of fouling and process control. PMID:28358349

  10. Motor Oil Classification Based on Time-Resolved Fluorescence

    OpenAIRE

    Mu, Taotao; Chen, Siying; Zhang, Yinchao; Guo, Pan; Chen, He; Meng, Fandong

    2014-01-01

    A time-resolved fluorescence (TRF) technique is presented for classifying motor oils. The system is constructed with a third harmonic Nd:YAG laser, a spectrometer, and an intensified charge coupled device (ICCD) camera. Steady-state and time-resolved fluorescence (TRF) measurements are reported for several motor oils. It is found that steady-state fluorescence is insufficient to distinguish the motor oil samples. Then contour diagrams of TRF intensities (CDTRFIs) are acquired to serve as uniq...

  11. Fluorogen-based reporters for fluorescence imaging: a review

    Science.gov (United States)

    Jullien, Ludovic; Gautier, Arnaud

    2015-12-01

    Fluorescence bioimaging has recently jumped into a new area of spatiotemporal resolution and sensitivity thanks to synergistic advances in both optical physics and probe/biosensor design. This review focuses on the recent development of genetically encodable fluorescent reporters that bind endogenously present or exogenously applied fluorogenic chromophores (so-called fluorogens) and activate their fluorescence. We highlight the innovative engineering and design that gave rise to these new natural and synthetic fluorescent reporters, and describe some of the emerging applications in imaging and biosensing.

  12. Normalized Quantitative Western Blotting Based on Standardized Fluorescent Labeling.

    Science.gov (United States)

    Faden, Frederik; Eschen-Lippold, Lennart; Dissmeyer, Nico

    2016-01-01

    Western blot (WB) analysis is the most widely used method to monitor expression of proteins of interest in protein extracts of high complexity derived from diverse experimental setups. WB allows the rapid and specific detection of a target protein, such as non-tagged endogenous proteins as well as protein-epitope tag fusions depending on the availability of specific antibodies. To generate quantitative data from independent samples within one experiment and to allow accurate inter-experimental quantification, a reliable and reproducible method to standardize and normalize WB data is indispensable. To date, it is a standard procedure to normalize individual bands of immunodetected proteins of interest from a WB lane to other individual bands of so-called housekeeping proteins of the same sample lane. These are usually detected by an independent antibody or colorimetric detection and do not reflect the real total protein of a sample. Housekeeping proteins-assumed to be constitutively expressed mostly independent of developmental and environmental states-can greatly differ in their expression under these various conditions. Therefore, they actually do not represent a reliable reference to normalize the target protein's abundance to the total amount of protein contained in each lane of a blot.Here, we demonstrate the Smart Protein Layers (SPL) technology, a combination of fluorescent standards and a stain-free fluorescence-based visualization of total protein in gels and after transfer via WB. SPL allows a rapid and highly sensitive protein visualization and quantification with a sensitivity comparable to conventional silver staining with a 1000-fold higher dynamic range. For normalization, standardization and quantification of protein gels and WBs, a sample-dependent bi-fluorescent standard reagent is applied and, for accurate quantification of data derived from different experiments, a second calibration standard is used. Together, the precise quantification of

  13. Evaluation of a radiative transfer equation and diffusion approximation hybrid forward solver for fluorescence molecular imaging.

    Science.gov (United States)

    Gorpas, Dimitris; Andersson-Engels, Stefan

    2012-12-01

    The solution of the forward problem in fluorescence molecular imaging strongly influences the successful convergence of the fluorophore reconstruction. The most common approach to meeting this problem has been to apply the diffusion approximation. However, this model is a first-order angular approximation of the radiative transfer equation, and thus is subject to some well-known limitations. This manuscript proposes a methodology that confronts these limitations by applying the radiative transfer equation in spatial regions in which the diffusion approximation gives decreased accuracy. The explicit integro differential equations that formulate this model were solved by applying the Galerkin finite element approximation. The required spatial discretization of the investigated domain was implemented through the Delaunay triangulation, while the azimuthal discretization scheme was used for the angular space. This model has been evaluated on two simulation geometries and the results were compared with results from an independent Monte Carlo method and the radiative transfer equation by calculating the absolute values of the relative errors between these models. The results show that the proposed forward solver can approximate the radiative transfer equation and the Monte Carlo method with better than 95% accuracy, while the accuracy of the diffusion approximation is approximately 10% lower.

  14. Determination of cadmium at ultra-trace levels by CPE-molecular fluorescence combined methodology

    Energy Technology Data Exchange (ETDEWEB)

    Talio, Maria Carolina [Instituto de Quimica de San Luis (INQUISAL-CONICET), Chacabuco y Pedernera, 5700 San Luis (Argentina); Luconi, Marta O. [Area de Quimica Analitica, Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis, San Luis (Argentina); Masi, Adriana N. [Area de Bromatologia- Ensayo y Valoracion de Medicamentos, Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis, San Luis (Argentina); Instituto de Quimica de San Luis (INQUISAL-CONICET), Chacabuco y Pedernera, 5700 San Luis (Argentina); Fernandez, Liliana P., E-mail: lfernand@unsl.edu.ar [Area de Quimica Analitica, Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis, San Luis (Argentina); Instituto de Quimica de San Luis (INQUISAL-CONICET), Chacabuco y Pedernera, 5700 San Luis (Argentina)

    2009-10-15

    A highly sensitive micelle-mediated extraction methodology for the preconcentration and determination of trace levels of cadmium by molecular fluorescence has been developed. Metal was complexed with o-phenanthroline (o-phen) and eosin (eo) at pH 7.6 in buffer Tris medium and quantitatively extracted into a small volume of surfactant-rich phase of PONPE 7.5 after centrifugating. The chemical variables affecting cloud point extraction (CPE) were evaluated and optimized. The RSD for six replicates of cadmium determinations at 0.84 {mu}g L{sup -1} level was 1.17%. The linearity range using the preconcentration system was between 2.79 x 10{sup -3} {mu}g L{sup -1} and 2.81 {mu}g L{sup -1} with a correlation coefficient of 0.99. Under the optimal conditions, it obtained a LOD of 8.38 x 10{sup -4} {mu}g L{sup -1} and LOQ of 2.79 x 10{sup -3} {mu}g L{sup -1}. The method presented good sensitivity and selectivity and was applied to the determination of trace amounts of cadmium in commercially bottled mineral water, tap water and water well samples with satisfactory results. The proposed method is an innovative application of CPE-luminescence to metal analysis comparable in sensitivity and accuracy with atomic spectroscopies.

  15. Direct reconstruction of pharmacokinetic parameters in dynamic fluorescence molecular tomography by the augmented Lagrangian method

    Science.gov (United States)

    Zhu, Dianwen; Zhang, Wei; Zhao, Yue; Li, Changqing

    2016-03-01

    Dynamic fluorescence molecular tomography (FMT) has the potential to quantify physiological or biochemical information, known as pharmacokinetic parameters, which are important for cancer detection, drug development and delivery etc. To image those parameters, there are indirect methods, which are easier to implement but tend to provide images with low signal-to-noise ratio, and direct methods, which model all the measurement noises together and are statistically more efficient. The direct reconstruction methods in dynamic FMT have attracted a lot of attention recently. However, the coupling of tomographic image reconstruction and nonlinearity of kinetic parameter estimation due to the compartment modeling has imposed a huge computational burden to the direct reconstruction of the kinetic parameters. In this paper, we propose to take advantage of both the direct and indirect reconstruction ideas through a variable splitting strategy under the augmented Lagrangian framework. Each iteration of the direct reconstruction is split into two steps: the dynamic FMT image reconstruction and the node-wise nonlinear least squares fitting of the pharmacokinetic parameter images. Through numerical simulation studies, we have found that the proposed algorithm can achieve good reconstruction results within a small amount of time. This will be the first step for a combined dynamic PET and FMT imaging in the future.

  16. Unraveling the molecular mechanism of photosynthetic toxicity of highly fluorescent silver nanoclusters to Scenedesmus obliquus.

    Science.gov (United States)

    Zhang, Li; Goswami, Nirmal; Xie, Jianping; Zhang, Bo; He, Yiliang

    2017-11-27

    While the discovery of numerous attractive properties of silver at the nanoscale has increased their demand in many sectors including medicine, optics, sensing, painting and cosmetics, it has also raised wide public concerns about their effect on living organisms in aquatic environment. Despite the continuous effort to understand the various aspects of the toxicity of silver nanomaterials, the molecular level understanding on their cytotoxicity mechanism to biological organisms has remained unclear. Herein, we demonstrated the underlying mechanism of the photosynthetic toxicity against green algae namely, Scenedesmus obliquus by using an emerging silver nanomaterial, called silver nanoclusters (defined as r-Ag NCs). By exploiting the unique fluorescence properties of r-Ag NCs along with various other analytical/biological tools, we proposed that the photosynthetic toxicity of r-Ag NCs was largely attributed to the "joint-toxicity" effect of particulate form of r-Ag NCs and its released Ag + , which resulted in the disruption of the electron transport chain of light reaction and affected the content of key enzymes (RuBP carboxylase/ oxygenase) of Calvin cycle of algae cells. We believe that the present study can also be applied to the assessment of the ecological risk derived from other metal nanoparticles.

  17. Development of a DMD-based fluorescence microscope

    NARCIS (Netherlands)

    Chakrova, N.; Rieger, B.; Stallinga, S.

    2015-01-01

    We present a versatile fluorescence microscope, built by complementing a conventional fluorescence microscope with a digital micro-mirror device (DMD) in the illumination path. Arbitrary patterns can be created on the DMD and projected onto the sample. This patterned illumination can be used to

  18. Progress on RNAi-based molecular medicines

    Directory of Open Access Journals (Sweden)

    Chen J

    2012-07-01

    Full Text Available Jing Chen, Jianping XieInstitute of Modern Biopharmaceuticals, State Key Laboratory Breeding Base of Ministry of Education Eco-Environment of the Three Gorges Reservoir Region, School of Life Sciences, Southwest University, Chongqing, ChinaAbstract: RNA interference (RNAi is a promising strategy to suppress the expression of disease-relevant genes and induce post-transcriptional gene silencing. Their simplicity and stability endow RNAi with great advantages in molecular medicine. Several RNAi-based drugs are in various stages of clinical investigation. This review summarizes the ongoing research endeavors on RNAi in molecular medicine, delivery systems for RNAi-based drugs, and a compendium of RNAi drugs in different stages of clinical development. Of special interest are RNAi-based drug target discovery and validation, delivery systems for RNAi-based drugs, such as nanoparticles, rabies virus protein-based vehicles, and bacteriophages for RNA packaging.Keywords: RNA interference, delivery systems, liposome, nanoparticle, molecular medicines

  19. Direct visualization of the reaction transformation and signal amplification in a DNA molecular machine with total internal reflection fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Rui eRen

    2013-10-01

    Full Text Available In this study, as a proof of concept, the signal amplification in an artificial DNA molecular machine was directly visualized via total internal reflection fluorescence microscopy (TIRFM. The molecular machine brought about obvious morphology change in DNA nanostructures as well as signal amplifications. On one hand, through a triggered and autonomically repeated RCA, a DNA nano-complex featuring a locked circular DNA template (serving as raw feed was converted into a long periodically repeated strand, i.e. the RCA products. Furthermore, this RCA was repeated in three controllable reaction phases, bring about progressive signal amplification. It was testified that the RCA products (presented as long thread-like fluorescent objects can be easily distinguished from the inputted DNA probes (presented as fluorescent dots, thus the transformation in reaction can be visualized. Also, by quantitive counting of the aforementioned fluorescence objects, the progress of the reaction through the phases, along with time, and over the lysozyme concentration can be demonstrated through TIRFM visualization. Overall, it was demonstrated that TIRFM is an efficient approach to quantitively visualize the biochemical processes at single-molecule level.

  20. A hydrogel based fluorescent micro array used for the characterization of liquid analytes

    International Nuclear Information System (INIS)

    Thete, A.R.; Henkel, T.; Goeckeritz, R.; Endlich, M.; Koehler, J.M.; Gross, G.A.

    2009-01-01

    A fluorimetric micro spot array using non-specific recognition function is described for the analysis of liquid samples. The array was composed of binary mixtures of various fluorescence dyes which were embedded in a hydrogel matrix. The interactions between the fluorescent dyes and their molecular surrounding inside the hydrogel, influence their fluorescence wave length and intensities. The array was used for the characterization of solvent mixtures. Developed fluorescence patterns of the complete array as well as the fluorescence intensity changes of single spots were analysed. It was proven, that specific analytical information can be gained using this non-specific recognition approach. The identification of some alcoholic beverages is described as an example of the application of this method when used for quality control purposes. Analogous to the appellation 'electronic nose' and 'electronic tongue' the described micro spot array acts as an 'optochemical tongue'

  1. A hydrogel based fluorescent micro array used for the characterization of liquid analytes

    Energy Technology Data Exchange (ETDEWEB)

    Thete, A.R. [Technical University of Ilmenau, Institute for Micro and Nano Technologies, Department of Physical Chemistry and Microreaction Technologies, Weimarerstr. 32, D-98693 Ilmenau (Germany)], E-mail: aniket.thete@tu-ilmenau.de; Henkel, T. [Institute for Photonic Technology, Department Microfluidic devices, Albert - Einstein-Street 9, D-07745 Jena (Germany); Goeckeritz, R.; Endlich, M.; Koehler, J.M.; Gross, G.A. [Technical University of Ilmenau, Institute for Micro and Nano Technologies, Department of Physical Chemistry and Microreaction Technologies, Weimarerstr. 32, D-98693 Ilmenau (Germany)

    2009-02-02

    A fluorimetric micro spot array using non-specific recognition function is described for the analysis of liquid samples. The array was composed of binary mixtures of various fluorescence dyes which were embedded in a hydrogel matrix. The interactions between the fluorescent dyes and their molecular surrounding inside the hydrogel, influence their fluorescence wave length and intensities. The array was used for the characterization of solvent mixtures. Developed fluorescence patterns of the complete array as well as the fluorescence intensity changes of single spots were analysed. It was proven, that specific analytical information can be gained using this non-specific recognition approach. The identification of some alcoholic beverages is described as an example of the application of this method when used for quality control purposes. Analogous to the appellation 'electronic nose' and 'electronic tongue' the described micro spot array acts as an 'optochemical tongue'.

  2. Optical materials based on molecular nano/microcrystals and ...

    Indian Academy of Sciences (India)

    Wintec

    Abstract. Methodologies that we developed recently for the fabrication of molecular crystals with size varia- tion in the nano to micro regime and polyelectrolyte templated mono and multilayer Langmuir–Blodgett films, are reviewed. The electronic absorption and strong fluorescence in the molecular nano/microcrystals are.

  3. Luminescent Metal-Organic-Framework-Based Label-Free Assay of Polyphenol Oxidase with Fluorescent Scan.

    Science.gov (United States)

    Li, Yue; Guo, An; Chang, Lan; Li, Wen-Juan; Ruan, Wen-Juan

    2017-05-11

    Metal-organic frameworks (MOFs) are emerging in recent years as a kind of versatile fluorescent sensing materials, but their application to enzyme assays has rarely been studied. Here, the first example of a MOF-based label-free enzyme assay system is reported. A luminescent MOF was synthesized and applied to the activity analysis of polyphenol oxidase (PPO). With its distinct responses to the phenolic substrate and o-quinone product, this MOF could transduce the extent of PPO-catalyzed oxidation to fluorescence signal and enable the real-time monitoring of this reaction. Wide substrate adaptability and high sensitivity (detection limit=0.00012 U mL -1 ) were exhibited by this method, which meets the requirement of common bioanalysis. Interestingly, by the comparison with molecular capturing reagents, the heterogeneous nature of this MOF-based assay effectively preventing the interaction with the enzyme was proven, thus ensuring the authenticity of results. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. SERS and fluorescence-based ultrasensitive detection of mercury in water.

    Science.gov (United States)

    Makam, Pandeeswar; Shilpa, Rohilla; Kandjani, Ahmad Esmaielzadeh; Periasamy, Selvakannan R; Sabri, Ylias Mohammad; Madhu, Chilakapati; Bhargava, Suresh Kumar; Govindaraju, Thimmaiah

    2018-02-15

    The development of reliable and ultrasensitive detection marker for mercury ions (Hg 2+ ) in drinking water is of great interest for toxicology assessment, environmental protection and human health. Although many Hg 2+ detection methods have been developed, only few offer sensitivities below 1pM. Herein, we describe a simple histidine (H) conjugated perylene diimide (PDI) bolaamphiphile (HPH) as a dual-responsive optical marker to develop highly selective and sensitive probe as visible (sol-to-gel transformation), fluorescence and SERS-based Hg 2+ sensor platform in the water. Remarkably, HPH as a SERS marker supported on Au deposited monodispersed nanospheres monolayers (Au-MNM) of polystyrene offers an unprecedented selectivity and the best ever reported detection limit (LOD) of 60 attomolar (aM, 0.01 parts-per-quadrillion (ppq)) for Hg 2+ in water. This is ten orders of magnitude lower than the United States Environmental Protection Agency (USEPA) tolerance limit of Hg 2+ in drinking water (10nM, 2 ppb). This simple and effective design principle of host-guest interactions driven fluorescence and SERS-based detection may inspire the future molecular engineering strategies for the development of ultrasensitive toxic analyte sensor platforms. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. DNA-functionalized gold nanoparticle-based fluorescence polarization for the sensitive detection of silver ions.

    Science.gov (United States)

    Wang, Gongke; Wang, Shuangli; Yan, Changling; Bai, Guangyue; Liu, Yufang

    2018-04-05

    Despite their practical applications, Ag + ions are environmental pollutants and affect human health. So the effective detection methods of Ag + ions are imperative. Herein, we developed a simple, sensitive, selective, and cost-effective fluorescence polarization sensor for Ag + detection in aqueous solution using thiol-DNA-functionalized gold nanoparticles (AuNPs). In this sensing strategy, Ag + ions can specifically interact with a cytosine-cytosine (CC) mismatch in DNA duplexes and form stable metal-mediated cytosine-Ag + -cytosine (C-Ag + -C) base pairs. The formation of the C-Ag + -C complex results in evident changes in the molecular volume and fluorescence polarization signal. To achieve our aims, we prepared two complementary DNA strands containing C-base mismatches (probe A: 5'-SH-A 10 -TACCACTCCTCAC-3' and probe B: 5'-TCCTCACCAGTCCTA-FAM-3'). The stable hybridization between probe A and probe B occurs with the formation of the C-Ag + -C complex in the presence of Ag + ions, leading to obvious fluorescence quenching in comparison to the system without AuNP enhancement. The assay can be used to identify nanomolar levels of Ag + within 6 min at room temperature, and has extremely high specificity for Ag + , even in the presence of higher concentrations of interfering metal ions. Furthermore, the sensor was successfully applied to the detection of Ag + ions in environmental water samples and showed excellent selectivity and high sensitivity, implying its promising application in the future. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. A fluorescence-quenching platform based on biomineralized hydroxyapatite from natural seashell and applied to cancer cell detection.

    Science.gov (United States)

    Zhang, Ying; Liu, Wei; Banks, Craig E; Liu, Fei; Li, Mao; Xia, Fan; Yang, Xiangliang

    2014-12-19

    As a typical biomineral, hydroxyapatite (HAp) is widely applied in bone implants and other related fields. However, the inherent nature of HAp can potentially be altered through restricting its fabrication conditions. Here, HAp fabricated by a hydrothermal treatment of pieces of natural seashell is demonstrated to have the capability of fluorescence quenching. To the best of the author's knowledge, this is the first time that this new property of HAp has been reported. Consequently, we assembled a fluorescence-quenching platform based on the biomineralized HAp substrate following a hydrothermal treatment and associated with a DNA molecular beacon and applied to cancer cell detection by the transformation from "OFF state" (fluorescence quenching) to "ON state" (fluorescence recovery). Herein, we found that the outer surface of HAp material after hydrothermal biomineralization for 5 days has considerable capability for both fluorescence quenching and recovery. These results may also have implications in the further detection of various targets such as cancer cells with other special surface antigens, significant biological small molecules or disease related microRNA, just by changing the sequence of the nucleic acid beacon according to the corresponding aptamer.

  7. A Hadoop-based Molecular Docking System

    Science.gov (United States)

    Dong, Yueli; Guo, Quan; Sun, Bin

    2017-10-01

    Molecular docking always faces the challenge of managing tens of TB datasets. It is necessary to improve the efficiency of the storage and docking. We proposed the molecular docking platform based on Hadoop for virtual screening, it provides the preprocessing of ligand datasets and the analysis function of the docking results. A molecular cloud database that supports mass data management is constructed. Through this platform, the docking time is reduced, the data storage is efficient, and the management of the ligand datasets is convenient.

  8. ALA-based fluorescent diagnosis of malignant oral lesions in the presence of bacterial porphyrin formation

    Science.gov (United States)

    Schleier, P.; Berndt, A.; Zinner, K.; Zenk, W.; Dietel, W.; Pfister, W.

    2006-02-01

    The aminolevulinic acid (5-ALA) -based fluorescence diagnosis has been found to be promising for an early detection and demarcation of superficial oral squamous cell carcinomas (OSCC). This method has previously demonstrated high sensitivity, however this clinical trial showed a specificity of approximately 62 %. This specificity was mainly restricted by tumor detection in the oral cavity in the presence of bacteria. After topical ALA application in the mouth of patients with previously diagnosed OSSC, red fluorescent areas were observed which did not correlate to confirm histological findings. Swabs and plaque samples were taken from 44 patients and cultivated microbiologically. Fluorescence was investigated (OMA-system) from 32 different bacteria strains found naturally in the oral cavity. After ALA incubation, 30 of 32 strains were found to synthesize fluorescent porphyrins, mainly Protoporphyrin IX. Also multiple fluorescent spectra were obtained having peak wavelengths of 636 nm and around 618 nm - 620 nm indicating synthesis of different porphyrins, such as the lipophylic Protoporphyrin IX (PpIX) and hydrophylic porphyrins (water soluble porphyrins, wsp). Of the 32 fluorescent bacterial strains, 18 produced wsp, often in combination with PpIX, and 5 produced solely wsp. These results clarify that ALA-based fluorescence diagnosis without consideration or suppression of bacteria fluorescence may lead to false-positive findings. It is necessary to suppress bacteria fluorescence with suitable antiseptics before starting the procedure. In this study, when specific antiseptic pre-treatment was performed bacterial associated fluorescence was significantly reduced.

  9. A "distorted-BODIPY"-based fluorescent probe for imaging of cellular viscosity in live cells.

    Science.gov (United States)

    Zhu, Hao; Fan, Jiangli; Li, Miao; Cao, Jianfang; Wang, Jingyun; Peng, Xiaojun

    2014-04-14

    Cellular viscosity is a critical factor in governing diffusion-mediated cellular processes and is linked to a number of diseases and pathologies. Fluorescent molecular rotors (FMRs) have recently been developed to determine viscosity in solutions or biological fluid. Herein, we report a "distorted-BODIPY"-based probe BV-1 for cellular viscosity, which is different from the conventional "pure rotors". In BV-1, the internal steric hindrance between the meso-CHO group and the 1,7-dimethyl group forced the boron-dipyrrin framework to be distorted, which mainly caused nonradiative deactivation in low-viscosity environment. BV-1 gave high sensitivity (x=0.62) together with stringent selectivity to viscosity, thus enabling viscosity mapping in live cells. Significantly, the increase of cytoplasmic viscosity during apoptosis was observed by BV-1 in real time. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Ultrathin inorganic molecular nanowire based on polyoxometalates

    Science.gov (United States)

    Zhang, Zhenxin; Murayama, Toru; Sadakane, Masahiro; Ariga, Hiroko; Yasuda, Nobuhiro; Sakaguchi, Norihito; Asakura, Kiyotaka; Ueda, Wataru

    2015-01-01

    The development of metal oxide-based molecular wires is important for fundamental research and potential practical applications. However, examples of these materials are rare. Here we report an all-inorganic transition metal oxide molecular wire prepared by disassembly of larger crystals. The wires are comprised of molybdenum(VI) with either tellurium(IV) or selenium(IV): {(NH4)2[XMo6O21]}n (X=tellurium(IV) or selenium(IV)). The ultrathin molecular nanowires with widths of 1.2 nm grow to micrometre-scale crystals and are characterized by single-crystal X-ray analysis, Rietveld analysis, scanning electron microscopy, X-ray photoelectron spectroscopy, ultraviolet–visible spectroscopy, thermal analysis and elemental analysis. The crystals can be disassembled into individual molecular wires through cation exchange and subsequent ultrasound treatment, as visualized by atomic force microscopy and transmission electron microscopy. The ultrathin molecular wire-based material exhibits high activity as an acid catalyst, and the band gap of the molecular wire-based crystal is tunable by heat treatment. PMID:26139011

  11. Eco-friendly carbon-nanodot-based fluorescent paints for advanced photocatalytic systems

    OpenAIRE

    Young Park, So; Uk Lee, Hyun; Lee, Young-Chul; Choi, Saehae; Hyun Cho, Dae; Sik Kim, Hee; Bang, Sunghee; Seo, Soonjoo; Chang Lee, Soon; Won, Jonghan; Son, Byung-Chul; Yang, Mino; Lee, Jouhahn

    2015-01-01

    Fluorescent carbon nanomaterials, especially zero-dimensional (0D) carbon nanodots (CDs), are widely used in broad biological and optoelectronic applications. CDs have unique characteristics such as strong fluorescence, biocompatibility, sun-light response, and capability of mass-production. Beyond the previous green CD obtained from harmful natural substances, we report a new type of fluid-based fluorescent CD paints (C-paints) derived from polyethylene glycol (PEG; via simple ultrasound irr...

  12. Base motif recognition and design of DNA templates for fluorescent silver clusters by machine learning.

    Science.gov (United States)

    Copp, Stacy M; Bogdanov, Petko; Debord, Mark; Singh, Ambuj; Gwinn, Elisabeth

    2014-09-03

    Discriminative base motifs within DNA templates for fluorescent silver clusters are identified using methods that combine large experimental data sets with machine learning tools for pattern recognition. Combining the discovery of certain multibase motifs important for determining fluorescence brightness with a generative algorithm, the probability of selecting DNA templates that stabilize fluorescent silver clusters is increased by a factor of >3. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Molecular Level Understanding of Sodium Dodecyl Sulfate (SDS) Induced Sol-Gel Transition of Pluronic F127 Using Fisetin as a Fluorescent Molecular Probe.

    Science.gov (United States)

    Mishra, Jhili; Swain, Jitendriya; Mishra, Ashok Kumar

    2018-01-11

    The thermoreversible sol-gel transition of pluronic F127 is markedly altered even with addition of submicellar concentration of sodium dodecyl sulfate (SDS) surfactant. Multiple fluorescence parameters like fluorescence intensity, fluorescence anisotropy and fluorescence lifetime of both the prototropic forms (anion (A - *) and phototautomer FT*) of the photoprototropic fluorescent probe fisetin has been efficiently used to understand the molecular level properties like polarity and microviscosity of the PF127-SDS system as a function of temperature. The SDS-induced increase in the interfacial hydrophobicity level is seen to affect the sol-gel phase transition of PF127 (21-18 °C). The E T (30) polarity parameter value of anionic emission of fisetin suggests that there is a considerable decrease in the polarity of the PF127 medium with increase in temperature and with the addition of SDS. The microviscosity progressively increases from ∼5 mPa s (sol state, 10 °C) to ∼22.01 mPa s (gel state 35 °C) in aqueous solution of PF127. The variation in microviscosity with addition of SDS in PF127-SDS mixed system is significant in sol phase whereas in gel phase this variation is significantly less. Temperature dependent fluorescence lifetime of FT* indicates that there is heterogeneity in distribution of fisetin molecules at different domains of PF127. This work also show-cases the sensitivity of fisetin toward change in polarity and change in sol-gel transition temperature of copolymer PF127 with variation in temperature (both forward and reverse directions) and SDS.

  14. Identification of adiponectin receptor agonist utilizing a fluorescence polarization based high throughput assay.

    Directory of Open Access Journals (Sweden)

    Yiyi Sun

    Full Text Available Adiponectin, the adipose-derived hormone, plays an important role in the suppression of metabolic disorders that can result in type 2 diabetes, obesity, and atherosclerosis. It has been shown that up-regulation of adiponectin or adiponectin receptor has a number of therapeutic benefits. Given that it is hard to convert the full size adiponectin protein into a viable drug, adiponectin receptor agonists could be designed or identified using high-throughput screening. Here, we report on the development of a two-step screening process to identify adiponectin agonists. First step, we developed a high throughput screening assay based on fluorescence polarization to identify adiponectin ligands. The fluorescence polarization assay reported here could be adapted to screening against larger small molecular compound libraries. A natural product library containing 10,000 compounds was screened and 9 hits were selected for validation. These compounds have been taken for the second-step in vitro tests to confirm their agonistic activity. The most active adiponectin receptor 1 agonists are matairesinol, arctiin, (--arctigenin and gramine. The most active adiponectin receptor 2 agonists are parthenolide, taxifoliol, deoxyschizandrin, and syringin. These compounds may be useful drug candidates for hypoadiponectin related diseases.

  15. Gas Sensors Based on Molecular Imprinting Technology.

    Science.gov (United States)

    Zhang, Yumin; Zhang, Jin; Liu, Qingju

    2017-07-04

    Molecular imprinting technology (MIT); often described as a method of designing a material to remember a target molecular structure (template); is a technique for the creation of molecularly imprinted polymers (MIPs) with custom-made binding sites complementary to the target molecules in shape; size and functional groups. MIT has been successfully applied to analyze; separate and detect macromolecular organic compounds. Furthermore; it has been increasingly applied in assays of biological macromolecules. Owing to its unique features of structure specificity; predictability; recognition and universal application; there has been exploration of the possible application of MIPs in the field of highly selective gas sensors. In this present study; we outline the recent advances in gas sensors based on MIT; classify and introduce the existing molecularly imprinted gas sensors; summarize their advantages and disadvantages; and analyze further research directions.

  16. Digital logic circuit based on two component molecular systems of BSA and salen

    Science.gov (United States)

    Hai-Bin, Lin; Feng, Chen; Hong-Xu, Guo

    2018-02-01

    A new fluorescent molecular probe 1 was designed and constructed by combining bovine serum albumin (BSA) and N,N‧-bis(salicylidene)ethylenediamine (salen). Stimulated by Zn2 +, tris, or EDTAH2Na2, the distance between BSA and salen was regulated, which was accompanied by an obvious change in the fluorescence intensity at 350 or 445 nm based on Förster resonance energy transfer. Moreover, based on the encoding binary digits in these inputs and outputs applying positive logic conventions, a monomolecular circuit integrating one OR, three NOT, and three YES gates, was successfully achieved.

  17. Optical detection of λ-cyhalothrin by core-shell fluorescent molecularly imprinted polymers in Chinese spirits.

    Science.gov (United States)

    Wang, Jixiang; Gao, Lin; Han, Donglai; Pan, Jianming; Qiu, Hao; Li, Hongji; Wei, Xiao; Dai, Jiangdong; Yang, Jinghai; Yao, Hui; Yan, Yongsheng

    2015-03-11

    In this study, fluorescent molecularly imprinted polymers (FMIPs), which were for the selective recognition and fluorescence detection of λ-cyhalothrin (LC), were synthesized via fluorescein 5(6)-isothiocyanate (FITC) and 3-aminopropyltriethoxysilane (APTS)/SiO2 particles. The SiO2@FITC-APTS@MIPs were characterized by Fourier transform infrared (FT-IR), UV-vis spectrophotometer (UV-vis), fluorescence spectrophotometer, thermogravimetric analysis (TGA), confocal laser scanning microscope (CLSM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The as-synthesized SiO2@FITC-APTS@MIPs with an imprinted polymer film (thickness was about 100 nm) was demonstrated to be spherically shaped and had good monodispersity, high fluorescence intensity, and good selective recognition. Using fluorescence quenching as the detection tool, the largest fluorescence quenching efficiency (F0/F - 1) of SiO2@FITC-APTS@MIPs is close to 2.5 when the concentration of the LC is 1.0 μM L(-1). In addition, a linear relationship (F0/F - 1= 0.0162C + 0.0272) could be obtained covering a wide concentration range of 0-60 nM L(-1) with a correlation coefficient of 0.9968 described by the Stern-Volmer equation. Moreover, the limit of detection (LOD) of the SiO2@FITC-APTS@MIPs was 9.17 nM L(-1). The experiment results of practical detection revealed that the SiO2@FITC-APTS@MIPs as an attractive recognition element was satisfactory for the determination of LC in Chinese spirits. Therefore, this study demonstrated the potential of SiO2@FITC-APTS@MIPs for the recognition and detection of LC in food.

  18. Recent progress in design of protein-based fluorescent biosensors and their cellular applications.

    Science.gov (United States)

    Tamura, Tomonori; Hamachi, Itaru

    2014-12-19

    Protein-based fluorescent biosensors have emerged as key bioanalytical tools to visualize and quantify a wide range of biological substances and events in vitro, in cells, and even in vivo. On the basis of the construction method, the protein-based fluorescent biosensors can be principally classified into two classes: (1) genetically encoded fluorescent biosensors harnessing fluorescent proteins (FPs) and (2) semisynthetic biosensors comprised of protein scaffolds and synthetic fluorophores. Recent advances in protein engineering and chemical biology not only allowed the further optimization of conventional biosensors but also facilitated the creation of novel biosensors based on unique strategies. In this review, we survey the recent studies in the development and improvement of protein-based fluorescent biosensors and highlight the successful applications to live cell and in vivo imaging. Furthermore, we provide perspectives on possible future directions of the technique.

  19. Modulation of a solid-state reversible fluorescent photoswitching based on a controllable photochromic pyrazolones

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Hu; Guo, Jixi [Key Laboratory of Material and Technology for Clean Energy, Ministry of Education, Key Laboratory of Advanced Functional Materials, Autonomous Region, Institute of Applied Chemistry, Xinjiang University, Urumqi 830046, Xinjiang (China); Jia, Dianzeng, E-mail: jdz@xju.edu.cn [Key Laboratory of Material and Technology for Clean Energy, Ministry of Education, Key Laboratory of Advanced Functional Materials, Autonomous Region, Institute of Applied Chemistry, Xinjiang University, Urumqi 830046, Xinjiang (China); Guo, Mingxi; Le, Fuhe; Liu, Lang; Wu, Dongling [Key Laboratory of Material and Technology for Clean Energy, Ministry of Education, Key Laboratory of Advanced Functional Materials, Autonomous Region, Institute of Applied Chemistry, Xinjiang University, Urumqi 830046, Xinjiang (China); Li, Feng [State Laboratory of Surface and Interface Science and Technology, School of Materials and Chemical Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, Henan (China); University of Texas, M. D. Anderson Cancer Center, Houston, TX 77002 (United States)

    2014-08-15

    A novel solid-state reversible fluorescence photoswitching system (FPS) based on photochromism of photochromic pyrazolones has been developed by employing phosphor Sr{sub 2}P{sub 2}O{sub 7} co-doped with europium ion and chlorine ion (Sr{sub 2}P{sub 2}O{sub 7}–EC) and 1,3-diphenyl-4-(3-chlorobenzal)-5-hydroxypyrazole-4-phenylsemicarbazone (1a) as the fluorescence dye and the photochromic compound, respectively. With carefully selected components, the absorption band of the keto-form photochromic pyrazolones well overlaps with the emission peak of Sr{sub 2}P{sub 2}O{sub 7}–EC. The fluorescence emission intensity of Sr{sub 2}P{sub 2}O{sub 7}–EC is efficiently modulated by the photoisomerization of 1a with controlling the exposure time in the solid state. The fluorescence photoswitching system displayed high fluorescence quenching efficiency and remarkable fatigue resistance. It can be repeated 7 cycles without observable the changes of emission intensity. A fluorescence quenching efficiency can be achieved with a reversible colour change from white to yellow. - Graphical abstract: A novel fluorescence photoswitching system based on doping inorganic fluorescence dye into photochromic pyrazolones was constructed successfully. Its fluorescence emission could be efficiently modulated by the photoisomerization of pyrazolones. - Highlights: • A solid-state fluorescence photoswitching material was prepared. • Photoswitching is due to energy transfer between pyrazolone and fluorescence dye. • It exhibits excellent fluorescence contrast and fatigue resistance in the solid state.

  20. Saturated virtual fluorescence emission difference microscopy based on detector array

    Science.gov (United States)

    Liu, Shaocong; Sun, Shiyi; Kuang, Cuifang; Ge, Baoliang; Wang, Wensheng; Liu, Xu

    2017-07-01

    Virtual fluorescence emission difference microscopy (vFED) has been proposed recently to enhance the lateral resolution of confocal microscopy with a detector array, implemented by scanning a doughnut-shaped pattern. Theoretically, the resolution can be enhanced by around 1.3-fold compared with that in confocal microscopy. For further improvement of the resolving ability of vFED, a novel method is presented utilizing fluorescence saturation for super-resolution imaging, which we called saturated virtual fluorescence emission difference microscopy (svFED). With a point detector array, matched solid and hollow point spread functions (PSF) can be obtained by photon reassignment, and the difference results between them can be used to boost the transverse resolution. Results show that the diffraction barrier can be surpassed by at least 34% compared with that in vFED and the resolution is around 2-fold higher than that in confocal microscopy.

  1. Determining the fate of fluorescent quantum dots on surface of engineered budding S. cerevisiae cell molecular landscape.

    Science.gov (United States)

    Chouhan, Raghuraj S; Qureshi, Anjum; Niazi, Javed H

    2015-07-15

    In this study, we surface engineered living S. cerevisiae cells by decorating quantum dots (QDs) and traced the fate of QDs on molecular landscape of single mother cell through several generation times (progeny cells). The fate of QDs on cell-surface was tracked through the cellular division events using confocal microscopy and fluorescence emission profiles. The extent of cell-surface QDs distribution among the offspring was determined as the mother cell divides into daughter cells. Fluorescence emission from QDs on progeny cells was persistent through the second-generation time (~240min) until all of the progeny cells lost their cell-bound QDs during the third generation time (~360min). The surface engineered yeast cells were unaffected by the QDs present on their molecular landscapes and retained their normal cellular growth, architecture and metabolic activities as confirmed by their viability, scanning electron microscopy (SEM) examinations and cytotoxicity tests, respectively. Our results demonstrated that QDs on mother cell landscape tend to distribute among its progeny cells that accompanied with concomitant reduction in QDs' fluorescence, which can be quantified. We suggest that surface engineered cells with QDs will enable investigating the cellular behavior and monitoring cell growth patterns as nanobiosensors for screening of drugs/chemicals at single cell level with fewer side effects. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Binding analysis for interaction of diacetylcurcumin with β-casein nanoparticles by using fluorescence spectroscopy and molecular docking calculations

    Science.gov (United States)

    Mehranfar, Fahimeh; Bordbar, Abdol-Khalegh; Fani, Najme; Keyhanfar, Mehrnaz

    2013-11-01

    The interaction of diacetylcurcumin (DAC), as a novel synthetic derivative of curcumin, with bovine β-casein (an abundant milk protein that is highly amphiphilic and self assembles into stable micellar nanoparticles in aqueous solution) was investigated using fluorescence quenching experiments, Forster energy transfer measurements and molecular docking calculations. The fluorescence quenching measurements revealed the presence of a single binding site on β-casein for DAC with the binding constant value equals to (4.40 ± 0.03) × 104 M-1. Forster energy transfer measurements suggested that the distance between bound DAC and Trp143 residue is higher than the respective critical distance, hence, the static quenching is more likely responsible for fluorescence quenching other than the mechanism of non-radiative energy transfer. Our results from molecular docking calculations indicated that binding of DAC to β-casein predominantly occurred through hydrophobic contacts in the hydrophobic core of protein. Additionally, in vitro investigation of the cytotoxicity of free DAC and DAC-β-casein complex in human breast cancer cell line MCF7 revealed the higher cytotoxic effect of DAC-β-casein complex.

  3. Excitation and deexcitation of N2 molecular levels. Induced fluorescence by electrons and laser

    International Nuclear Information System (INIS)

    Perez Fernandez-Mayoralas, A.

    1989-01-01

    The electron impact excitation followed by fluorescence induced by N 2 -laser absorption was used to study the lifetime of the lowest vibrational level of the B 3 π g electronic state of N 2 . The experimental result of this work is 13 + 1 μs. To measure the lifetime of B 3 π g (v=2,3,5,6,7,8) levels the delayed coincidence method by electron impact was use. The lifetime values were compared with recent experimental and theoretical results. The relative intensi-ties of 3 π g --- A 3 Σ Ω + system bands, in the range (6540-10500 A o ) was measured using a hollow cathode lamp as spectral source. The relative transition moments and its dependence versus the r-centroid was obtained. Total cross sections for electron scattering by N molecules in the range 600 - 5000 eV have been obtained from measurements of the attenuation of a linear electron beam. The results have been compared with available experimental cross sections and with theoretical calculations based on the first Born approximation. (Author)

  4. Molecular engineering of a fluorescent bioprobe for sensitive and selective detection of amphibole asbestos.

    Directory of Open Access Journals (Sweden)

    Takenori Ishida

    Full Text Available Fluorescence microscopy-based affinity assay could enable highly sensitive and selective detection of airborne asbestos, an inorganic environmental pollutant that can cause mesothelioma and lung cancer. We have selected an Escherichia coli histone-like nucleoid structuring protein, H-NS, as a promising candidate for an amphibole asbestos bioprobe. H-NS has high affinity to amphibole asbestos, but also binds to an increasingly common asbestos substitute, wollastonite. To develop a highly specific Bioprobe for amphibole asbestos, we first identified a specific but low-affinity amosite-binding sequence by slicing H-NS into several fragments. Second, we constructed a streptavidin tetramer complex displaying four amosite-binding fragments, resulting in the 250-fold increase in the probe affinity as compared to the single fragment. The tetramer probe had sufficient affinity and specificity for detecting all the five types of asbestos in the amphibole group, and could be used to distinguish them from wollastonite. In order to clarify the binding mechanism and identify the amino acid residues contributing to the probe's affinity to amosite fibers, we constructed a number of shorter and substituted peptides. We found that the probable binding mechanism is electrostatic interaction, with positively charged side chains of lysine residues being primarily responsible for the probe's affinity to asbestos.

  5. Fluorescence-based biosensors from concepts to applications

    CERN Document Server

    Morris, May C

    2013-01-01

    One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors. Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed a

  6. Ruthenium(II) complex-based fluorescent sensor for peroxynitrite

    Science.gov (United States)

    Ma, Jingjin; Wu, Jiasheng; Liu, Weimin; Wang, Pengfei; Fan, Zhiyuan

    We have developed a new ruthenium complex, Ru(bpy)2[4-(2,2'-bipyridin-4-yloxy)phenol]Cl2 (RuL), as a fluorescent sensor to detect peroxynitrite (ONOO-). The results showed that the addition of ONOO- to the aqueous solution of RuL would result in distinct fluorescence quenching at 600 nm. RuL exhibits a good selectivity for ONOO- over other reactive oxygen species (ROS) and reactive nitrite species (RNS), and the reaction time is less than 1.5 s. The sensing mechanism is proposed as the oxidative O-dealkylation reaction.

  7. Molecular phylogeny of Ranunculaceae based on internal ...

    African Journals Online (AJOL)

    The botanical family Ranunculaceae contains important medicinal plants. To obtain new evolutionary evidence regarding the systematic classification of Ranunculaceae plants, we used molecular phylogenies to test relationships based on the internal transcribed spacer region. The results of phylogenetic analysis of 92 ...

  8. Molecular phylogeny of Ranunculaceae based on internal ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... The botanical family Ranunculaceae contains important medicinal plants. To obtain new evolutionary evidence regarding the systematic classification of Ranunculaceae plants, we used molecular phylogenies to test relationships based on the internal transcribed spacer region. The results of phylogenetic ...

  9. Environmental Phosphorus Recovery Based on Molecular Bioscavengers

    DEFF Research Database (Denmark)

    Gruber, Mathias Felix

    -scale quantum calculations to macro-scale fluid simulations, are employed to hint at the potential of a recovery technology based on molecular bioscavengers. As a first approach, data mining is used to obtain statistical information about how proteins in nature interact with phosphate groups, thereby revealing...

  10. Simultaneous determination of acetylsalicylic acid, paracetamol and caffeine using solid-phase molecular fluorescence and parallel factor analysis.

    Science.gov (United States)

    Alves, Julio Cesar L; Poppi, Ronei J

    2009-05-29

    This paper describes the determination of acetylsalicylic acid (ASA), paracetamol and caffeine in pharmaceutical formulations using solid-phase molecular fluorescence and second order multivariate calibration. This methodology is applicable even in the presence of unknown interferences and with spectral overlap of the components in the mixture. Parallel factor analysis (PARAFAC) was used for model development, whose effectiveness was demonstrated by analysis of variance (ANOVA). Errors below 10% were obtained for all compounds using an external validation set. Benefits of the new procedures not included in the reference methods such as low cost, no need of sample preparation, simple and fast analysis using fluorescence spectrometer and no generation of waste, make this method very attractive, allowing for the simultaneous determination of compounds with good reproducibility and accuracy.

  11. Graphene-based nanoprobes for molecular diagnostics.

    Science.gov (United States)

    Chen, Shixing; Li, Fuwu; Fan, Chunhai; Song, Shiping

    2015-10-07

    In recent years, graphene has received widespread attention owing to its extraordinary electrical, chemical, optical, mechanical and structural properties. Lately, considerable interest has been focused on exploring the potential applications of graphene in life sciences, particularly in disease-related molecular diagnostics. In particular, the coupling of functional molecules with graphene as a nanoprobe offers an excellent platform to realize the detection of biomarkers, such as nucleic acids, proteins and other bioactive molecules, with high performance. This article reviews emerging graphene-based nanoprobes in electrical, optical and other assay methods and their application in various strategies of molecular diagnostics. In particular, this review focuses on the construction of graphene-based nanoprobes and their special advantages for the detection of various bioactive molecules. Properties of graphene-based materials and their functionalization are also comprehensively discussed in view of the development of nanoprobes. Finally, future challenges and perspectives of graphene-based nanoprobes are discussed.

  12. A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research

    Directory of Open Access Journals (Sweden)

    Alexander Gust

    2014-09-01

    Full Text Available Single-molecule fluorescence techniques are ideally suited to provide information about the structure-function-dynamics relationship of a biomolecule as static and dynamic heterogeneity can be easily detected. However, what type of single-molecule fluorescence technique is suited for which kind of biological question and what are the obstacles on the way to a successful single-molecule microscopy experiment? In this review, we provide practical insights into fluorescence-based single-molecule experiments aiming for scientists who wish to take their experiments to the single-molecule level. We especially focus on fluorescence resonance energy transfer (FRET experiments as these are a widely employed tool for the investigation of biomolecular mechanisms. We will guide the reader through the most critical steps that determine the success and quality of diffusion-based confocal and immobilization-based total internal reflection fluorescence microscopy. We discuss the specific chemical and photophysical requirements that make fluorescent dyes suitable for single-molecule fluorescence experiments. Most importantly, we review recently emerged photoprotection systems as well as passivation and immobilization strategies that enable the observation of fluorescently labeled molecules under biocompatible conditions. Moreover, we discuss how the optical single-molecule toolkit has been extended in recent years to capture the physiological complexity of a cell making it even more relevant for biological research.

  13. FRET Between Riboflavin and 9-Anthraldehyde Based Fluorescent Organic Nanoparticles Possessing Antibacterial Activity.

    Science.gov (United States)

    Mahajan, Prasad G; Dige, Nilam C; Suryawanshi, Sonali B; Dalavi, Dattatray K; Kamble, Avinash A; Bhopate, Dhanaji P; Kadam, Abhijit N; Kondalkar, Vijay V; Kolekar, Govind B; Patil, Shivajirao R

    2018-01-01

    The aqueous suspension of fluorescent nanoparticles were prepared by using 9-anthradehdye derivative (AH). The nanoparticles (AHNPs) were characterized using DLS-zeta sizer and SEM techniques. The photo physical properties of nanoparticles and precursor were measured and compared using UV-absorption spectroscopy, fluorescence spectroscopy and fluorescence lifetime studies. The significant overlap between fluorescence spectrum of AHNPs and excitation spectrum of Riboflavin (RF) led us to explore Fluorescence Resonance Energy Transfer (FRET) studies between AHNPs and RF in aqueous medium. The mechanism of FRET from AHNPs to RF discussed on spectral observations, thermodynamic parameters and changes produces in fluorescence lifetime in absence and presence of different concentrations of RF to AHNPs. The limit of detection for RF (0.071 µM) is considerably low compared with reported methods. Thus, we explore AHNPs as novel nano probe for quantitative determination of RF in pharmaceutical samples based on FRET study. In addition with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture viz. E. coli and Bacillus sps. Graphical Abstract 9-anthradehdye based fluorescent nanoparticles (AHNPs) explores as nano probe to detect Riboflavin (RF) in aqueous medium based on Fluorescence Resonance Energy Transfer (FRET) studies. The proposed analytical method successfully applied for quantitative determination of RF in pharmaceutical samples. In addition, with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture suspension viz. E. coli and Bacillus sps.

  14. A quinoline based pH sensitive ratiometric fluorescent sensor ...

    Indian Academy of Sciences (India)

    Fluorescence measurements were performed on a Hitachi F7000 spectrofluorimeter. Mass spectra measurement was carried out by Waters Xevo G2-. S QTof Mass Spectrometer. pH measurements were carried out using Orion 3-Star Plus pH Benchtop Meter. 2.2 Synthesis. The compound 1 was synthesized by stirring an.

  15. A coumarin-based colorimetric fluorescent probe for hydrogen sulfide

    Indian Academy of Sciences (India)

    H2S) is presented. This 'off–on' probe exhibited high selectivity ... beneficial for the design of fluorescence sensors, which result in ON-OFF, OFF-ON or ratiometric ... The initial nucleophilic attack of H2S would lead to an intermediate thiol, which ...

  16. The fast polarization modulation based dualfocus fluorescence correlation spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Štefl, Martin; Benda, Aleš; Gregor, I.; Hof, Martin

    2014-01-01

    Roč. 22, č. 1 (2014), s. 885-899 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

  17. Development of a Green Fluorescent Protein-Based Laboratory Curriculum

    Science.gov (United States)

    Larkin, Patrick D.; Hartberg, Yasha

    2005-01-01

    A laboratory curriculum has been designed for an undergraduate biochemistry course that focuses on the investigation of the green fluorescent protein (GFP). The sequence of procedures extends from analysis of the DNA sequence through PCR amplification, recombinant plasmid DNA synthesis, bacterial transformation, expression, isolation, and…

  18. Study on serum fluorescence spectra based on wavelet transform

    African Journals Online (AJOL)

    STORAGESEVER

    2010-02-08

    Feb 8, 2010 ... index of blood, hence providing help for early diagnosis and cure for diseases. ... as mathematics, physics, image processing and spec- trum analysis (Qin et al., 2006; Wang and Li, 2006; ... The blood serum fluorescence spectrum was detected by using the system shown in Figure 1. The equipment used ...

  19. Study on serum fluorescence spectra based on wavelet transform ...

    African Journals Online (AJOL)

    Blood plays an important role in clinical diagnosis and treatment and as such, the analysis of blood spectrum will be of very important practical significance. Serum fluorescence emission intensity is closely related with the excitation wavelength; when the excitation wavelength is 230 nm, the blood lipid concentration and ...

  20. Orthonormal Wavelet Bases for Quantum Molecular Dynamics

    International Nuclear Information System (INIS)

    Tymczak, C.; Wang, X.

    1997-01-01

    We report on the use of compactly supported, orthonormal wavelet bases for quantum molecular-dynamics (Car-Parrinello) algorithms. A wavelet selection scheme is developed and tested for prototypical problems, such as the three-dimensional harmonic oscillator, the hydrogen atom, and the local density approximation to atomic and molecular systems. Our method shows systematic convergence with increased grid size, along with improvement on compression rates, thereby yielding an optimal grid for self-consistent electronic structure calculations. copyright 1997 The American Physical Society

  1. Interactions between epinastine and human serum albumin: Investigation by fluorescence, UV-vis, FT-IR, CD, lifetime measurement and molecular docking

    Science.gov (United States)

    Ariga, Girish G.; Naik, Praveen N.; Chimatadar, Shivamurti A.; Nandibewoor, Sharanappa T.

    2017-06-01

    The fluorescence quenching of human serum albumin (HSA) by epinastine hydrochloride (EPN) at pH 7.4 buffer was studied using absorption, fluorescence quenching, time-resolved, circular-dichroism, synchronous and molecular docking studies have been employed in the system. The fluorescence quenching study revealed that the static quenching mechanism was involved in the interaction of EPN with human serum albumin. The value number of binding sites, n, is close to unity, EPN-HSA, indicated the presence of a single class of binding site for the drug in protein. The binding constant value of EPN_HSA was observed to be 2.72 × 104 M-1 at 298 K. The spectral results attest that the binding of EPN-HSA induced conformational changes in the HSA. The metal ions viz., Ca2+, Co2+, Cu2+, Ni2+ and Zn2+ were found to influence the binding of the EPN to HSA. Based on the Forster's theory of non-radiation energy transfer, the binding average distance, r, between the donor (HSA) and acceptor (EPN) was found to be 4.33 nm. The circular dichroism data revealed that the presence of EPN decreased the α-helix content of serum albumin, which indicated conformation changes in HSA upon interaction with EPN.

  2. Gold and silver nanoparticles based superquenching of fluorescence: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, Debanjana; Chattopadhyay, Nitin, E-mail: nitin.chattopadhyay@yahoo.com

    2015-04-15

    The short review highlights the recent advances on the gold and silver nanoparticles induced efficient quenching of fluorescence from various fluorophores looking at their promising use as optical rulers and chemo-/bio- sensors. The fluorescence quenching often leads to the increase in the Stern–Volmer constant (K{sub SV}~10{sup 7}–10{sup 10} mol{sup −1} dm{sup 3}) several orders of magnitude higher than the values observed for the normal photochemical quenching processes (~10{sup 2} mol{sup −1} dm{sup 3}). This amplified quenching has been termed as “super-quenching” or “hyper-quenching”. Energy transfer (ET) is established from the donor to the metal nanoparticles rationalizing these fast quenching processes. Considering the distance dependence of the ET process, Förster resonance energy transfer (FRET) and nanometal surface energy transfer (NSET) are ascribed to take place. These sensitive distance dependent phenomena serve as the spectroscopic ruler to measure the intra- or intermolecular distances between the interacting partners. In this account focus has been laid on the size dependent energy transfer and super- and hyper- quenching of the fluorescence of the donor moieties by the nanometals and their probable applications in sensing. Rationalization has been made for the nanoparticle induced huge enhancement in the quenching efficiency. The impact of this review lies in the possible application of these amplified quenching processes in designing high sensitive chemical and biological sensors. - Highlights: • Super efficient quenching of fluorescence of probes by gold and silver nanoparticles is highlighted. • The amplified fluorescence quenching of dyes and polymers is rationalized. • Energy transfer is assigned to be responsible for the efficient quenching process. • Amplified quenching has its potential use in designing sensitive chemical/biological sensors.

  3. Highly Sensitive Fluorescent Sensor for Cartap Based on Fluorescence Resonance Energy Transfer Between Gold Nanoparticles and Rhodamine B.

    Science.gov (United States)

    Dong, Liang; Hou, Changjun; Fa, Huanbao; Yang, Mei; Wu, Huixiang; Zhang, Liang; Huo, Danqun

    2018-04-01

    Cartap residue poses a great threat to human health and its derivatives would remain in soils, natural waters and other environmental domains for a long time. Herein, a simple, rapid and ultrasensitive analytical method for the determination of cartap based on fluorescence resonance energy transfer (FRET) between Au nanoparticles (AuNPs) and rhodamine B (RB) is first described. With the presence of citrate-stabilized AuNPs, the fluorescence of RB was remarkably quenched by AuNPs via FRET. The fluorescence of the AuNPs-RB system was recovered upon addition of cartap, cartap can be adsorbed on the surface of AuNPs due to its amino group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the FRET between AuNPs and RB was weakened and the PL intensity of RB was recovered accordingly. A good linear correlation for detection of RB was exhibited from 1 nM to 180 nM, and the detection limit reached 0.88 nM, which was much lower than the safety limit required by USA, UK and China. To the best of our knowledge, it has been the lowest detection ever without the aid of costly instrumentation. This method was successfully carried out for the assessment of cartap in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost and non-time-consuming compared with traditional methods.

  4. Investigation of the interaction between isomeric derivatives and human serum albumin by fluorescence spectroscopy and molecular modeling

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruiyong, E-mail: wangry@zzu.edu.cn; Dou, Huanjing; Yin, Yujing; Xie, Yuanzhe; Sun, Li; Liu, Chunmei; Dong, Jingjing; Huang, Gang; Zhu, Yanyan; Song, Chuanjun, E-mail: chjsong@zzu.edu.cn; Chang, Junbiao, E-mail: changjunbiao@zzu.edu.cn

    2014-10-15

    In this paper, we have synthesized 9H-pyrrolo[1,2-a]indol-9-ones and the isomeric indeno[2,1-b]pyrrol-8-ones. The interactions of human serum albumin with series of isomeric derivatives have been studied by spectrophotometric methods. Results show the intrinsic fluorescence is quenched by the derivatives with a static quenching procedure. The thermodynamics parameters indicate that van der Waals forces and hydrogen bonds play a major role in the interactions. The results of synchronous fluorescence spectra demonstrate that the microenvironments of Trp residue of human serum albumin are disturbed by most derivatives. Thermodynamic results showed that the 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers and bind to human serum albumin with the higher affinity than isomeric indeno[2,1-b]pyrrol-8-ones. The influence of molecular structure on the binding aspects has been investigated. - Highlights: • The interactions between isomeric derivatives and HSA have been investigated. • Results reveal that 9H-pyrrolo[1,2-a]indol-9-ones are stronger quenchers for HSA. • Hydrogen bonds and van der Waals forces play major role in the binding process. • The influence of molecular structure on the binding aspects has been investigated. • The binding study was also modeled by molecular docking.

  5. A nanoplasmonic switch based on molecular machines

    KAUST Repository

    Zheng, Yue Bing

    2009-06-01

    We aim to develop a molecular-machine-driven nanoplasmonic switch for its use in future nanophotonic integrated circuits (ICs) that have applications in optical communication, information processing, biological and chemical sensing. Experimental data show that an Au nanodisk array, coated with rotaxane molecular machines, switches its localized surface plasmon resonances (LSPR) reversibly when it is exposed to chemical oxidants and reductants. Conversely, bare Au nanodisks and disks coated with mechanically inert control compounds, do not display the same switching behavior. Along with calculations based on time-dependent density functional theory (TDDFT), these observations suggest that the nanoscale movements within surface-bound "molecular machines" can be used as the active components in plasmonic devices. ©2009 IEEE.

  6. A matter of collection and detection for intraoperative and noninvasive near-infrared fluorescence molecular imaging: To see or not to see?

    Science.gov (United States)

    Zhu, Banghe; Rasmussen, John C.; Sevick-Muraca, Eva M.

    2014-01-01

    Purpose: Although fluorescence molecular imaging is rapidly evolving as a new combinational drug/device technology platform for molecularly guided surgery and noninvasive imaging, there remains no performance standards for efficient translation of “first-in-humans” fluorescent imaging agents using these devices. Methods: The authors employed a stable, solid phantom designed to exaggerate the confounding effects of tissue light scattering and to mimic low concentrations (nM–pM) of near-infrared fluorescent dyes expected clinically for molecular imaging in order to evaluate and compare the commonly used charge coupled device (CCD) camera systems employed in preclinical studies and in human investigational studies. Results: The results show that intensified CCD systems offer greater contrast with larger signal-to-noise ratios in comparison to their unintensified CCD systems operated at clinically reasonable, subsecond acquisition times. Conclusions: Camera imaging performance could impact the success of future “first-in-humans” near-infrared fluorescence imaging agent studies. PMID:24506637

  7. A Thiazole Coumarin (TC) Turn-On Fluorescence Probe for AT-Base Pair Detection and Multipurpose Applications in Different Biological Systems

    Science.gov (United States)

    Narayanaswamy, Nagarjun; Kumar, Manoj; Das, Sadhan; Sharma, Rahul; Samanta, Pralok K.; Pati, Swapan K.; Dhar, Suman K.; Kundu, Tapas K.; Govindaraju, T.

    2014-09-01

    Sequence-specific recognition of DNA by small turn-on fluorescence probes is a promising tool for bioimaging, bioanalytical and biomedical applications. Here, the authors report a novel cell-permeable and red fluorescent hemicyanine-based thiazole coumarin (TC) probe for DNA recognition, nuclear staining and cell cycle analysis. TC exhibited strong fluorescence enhancement in the presence of DNA containing AT-base pairs, but did not fluoresce with GC sequences, single-stranded DNA, RNA and proteins. The fluorescence staining of HeLa S3 and HEK 293 cells by TC followed by DNase and RNase digestion studies depicted the selective staining of DNA in the nucleus over the cytoplasmic region. Fluorescence-activated cell sorting (FACS) analysis by flow cytometry demonstrated the potential application of TC in cell cycle analysis in HEK 293 cells. Metaphase chromosome and malaria parasite DNA imaging studies further confirmed the in vivo diagnostic and therapeutic applications of probe TC. Probe TC may find multiple applications in fluorescence spectroscopy, diagnostics, bioimaging and molecular and cell biology.

  8. Fluorescence Behavior and Dural Infiltration of Meningioma Analyzed by 5-Aminolevulinic Acid-Based Fluorescence: Operating Microscope Versus Mini-Spectrometer.

    Science.gov (United States)

    Knipps, Johannes; Beseoglu, Kerim; Kamp, Marcel; Fischer, Igor; Felsberg, Joerg; Neumann, Lisa M; Steiger, Hans-Jakob; Cornelius, Jan F

    2017-12-01

    To compare fluorescence intensity of tumor specimens, as measured by a fluorescence-guided surgery microscope and a spectrometer, to evaluate tumor infiltration of dura mater around meningiomas with help of these 2 different 5-aminolevulinic acid (5-ALA)-based fluorescence tools, and to correlate fluorescence intensity with histopathologic data. In a clinical series, meningiomas were resected by 5-ALA fluorescence-guided surgery. Fluorescence intensity was semiquantitatively rated by the surgeon at predefined points. Biopsies were harvested and fluorescence intensity measured by a spectrometer and histopathologically analyzed. Sampling was realized at the level of the dura in a centrifugal direction. A total of 104 biopsies (n = 13 tumors) were analyzed. Specificity and sensitivity of the microscope were 0.96 and 0.53 and of the spectrometer 0.95 and 0.93, respectively. Fluorescence intensity as measured by the spectrometer was correlated to histologically confirmed tumor burden. In a centrifugal direction, tumor burden and fluorescence intensity continuously decreased (along the dural tail). Below a threshold value of 639 arbitrary units no tumor was histologically detectable. At the level of the dura the spectrometer was highly sensitive for detection of meningioma cells. The surgical microscope showed false negative results and missed residual tumor cells in more than one half of the cases. The complementary use of both fluorescence tools may improve resection quality. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. A capillary-based probe for in situ detection of enhanced fluorescence signals

    Science.gov (United States)

    Long, F.; Xiao, R.; Zhu, A. N.; Shi, H. C.; Wang, S. Q.

    2013-07-01

    A simple, compact, and high sensitivity capillary-based probe for the in situ detection of fluorescence signals with high sensitivity is demonstrated. A home-made single-multi-mode fiber coupler that is coaxially aligned with the capillary-based probe provides for the transmission of excitation light and the collection and transmission of fluorescence. We propose a conceptually straightforward theoretical model to optimize the factors affecting the fluorescence-capture capability of the capillary-based probe. The fluorescence signal detected by fiber-optic spectroscopy non-linearly increases with the length of the capillary-based probe. In addition, the thicker the capillary tube wall is, the less the fluorescence signals determined are. The performance of the proposed probe is evaluated experimentally by measuring the fluorescence spectra of Cy5.5 dye and blue-green algae. The experimental results show that the proposed probe provides more than a ten-fold increase in fluorescence signal compared with direct measurements by a flat-tipped multi-mode fiber probe. The advantages of the capillary-based probe, which include its simple and compact structure, excellent light collection efficiency, requirement of small sample volume, and recoverability of samples, allow its wide application to in situ detection in the medical, forensic, biological, geological, and environmental fields with high sensitivity.

  10. A graphitic carbon nitride based fluorescence resonance energy transfer detection of riboflavin.

    Science.gov (United States)

    Han, Jing; Zou, Hong Yan; Gao, Ming Xuan; Huang, Cheng Zhi

    2016-01-01

    Fluorescence resonance energy transfer (FRET), which occurs between two luminescent chromophores, can greatly improve the selectivity and sensitivity of a fluorescent assay when a ratiometric signaling with the fluorescence enhancement of the acceptor at the expense of the donor is adopted. In this study, a fluorescence ratiometric detection (FRD) of riboflavin (RF) has been made based on FRET, as the strong overlap occurred between the emission spectrum of graphitic carbon nitride (g-C3N4) and absorption spectrum of RF, in which g-C3N4 acts as the energy donor and RF as the energy acceptor. With increasing concentration of RF, the fluorescence intensity of g-C3N4 emission at 444 nm decreased and the fluorescence peak at 523 nm for RF increased regularly, making the fluorescence intensity ratio of 523 nm to 444 nm linearly dependent on the concentration of RF in the range from 0.4 μM to 10 μM, giving a limit of the detection of 170 nM. This method can be used to quantify RF in complex systems such as milk and drink, showing that the novel FRET-based fluorescence ratiometric detection can enable an attractive assay platform for analytes of interest. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Visual and fluorescent detection of acetamiprid based on the inner filter effect of gold nanoparticles on ratiometric fluorescence quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Xu [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China); Li, Hongxia [School of Pharmacy, Jilin University, Changchun 130021 (China); Li, Yang [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China); Su, Xingguang, E-mail: suxg@jlu.edu.cn [Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012 (China)

    2014-12-10

    Highlights: • The RF-QDs were fabricated by two different QDs using layer-by-layer assembly methods. • The PL intensity of RF-QDs could be quenched by AuNPs based on inner-filter effect. • Acetamiprid can adsorb on AuNPs led to the PL intensity of RF-QDs recover properly. • AuNPs serve a dual function as fluorescence quencher and colorimetric reporter in the sensor. - Abstract: In this work, we develop a simple and rapid sensing method for the visual and fluorescent detection of acetamiprid (AC) based on the inner-filter effect (IFE) of gold nanoparticles (AuNPs) on ratiometric fluorescent quantum dots (RF-QDs). The RF-QDs based dual-emission nanosensor was fabricated by assembling green emissive QDs (QDs{sub 539} {sub nm}, λ{sub em} = 539 nm) on the surface of red emissive QDs (QDs{sub 661} {sub nm}, λ{sub em} = 661 nm)-doped silica microspheres. The photoluminescence (PL) intensity of RF-QDs could be quenched by AuNPs based on IFE. Acetamiprid can adsorb on the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on RF-QDs was weakened and the PL intensity of RF-QDs was recovered accordingly. Under the optimized conditions, the PL intensity of the RF-QDs/AuNPs system was proportional to the concentration of AC in the range of 0.025–5.0 μg mL{sup −1}, with a detection limit of 16.8 μg L{sup −1}. The established method had been used for AC detection in environmental and agricultural samples with satisfactory results.

  12. A Fluorescence Based Miniaturized Detection Module for Toxin Producing Algae

    Science.gov (United States)

    Zieger, S. E.; Mistlberger, G.; Troi, L.; Lang, A.; Holly, C.; Klimant, I.

    2016-12-01

    Algal blooms are sensitive to external environmental conditions and may pose a serious threat to marine and human life having an adverse effect on the ecosystem. Harmful algal blooms can produce different toxins, which can lead to massive fish kills or to human disorders. Facing these problems, miniaturized and low-cost instrumentation for an early detection and identification of harmful algae classes has become more important over the last years. 1,2Based on the characteristic pigment pattern of different algae classes, we developed a miniaturized detection module, which is able to detect and identify algae classes after analyzing their spectral behavior. Our device combines features of a flow-cytometer and fluorimeter and is build up as a miniaturized and low-cost device of modular design. Similar to a fluorimeter, it excites cells in the capillary with up to 8 different excitation wavelengths recording the emitted fluorescence at 4 different emission channels. Furthermore, the device operates in a flow-through mode similar to a flow-cytometer, however, using only low-cost elements such as LEDs and photodiodes. Due to its miniaturized design, the sensitivity and selectivity increase, whereas background effects are reduced. With a sampling frequency of 140 Hz, we try to detect and count particular cell events even at a concentration of 2 cells / 7.3 µL illuminated volume. Using a self-learning multivariate algorithm, the data are evaluated autonomously on the device enabling an in-situ analysis. The flexibility in choosing excitation and emission wavelengths as well as the high sampling rate enables laboratory applications such as measuring induction kinetics. However, in its first application, the device is part of an open and modular monitoring system enabling the sensing of chemical compounds such as toxic and essential Hg, Cd, Pb, As and Cu trace metal species, nutrients and species related to the carbon cycle, VOCs and potentially toxic algae classes (FP7

  13. A search for molecular hydrogen fluorescence near 100 km. [excitation by solar extreme UV radiation

    Science.gov (United States)

    Feldman, P. D.; Takacs, P. Z.

    1975-01-01

    The fluorescence of H2 in the Lyman band system, excited by solar extreme ultraviolet radiation, provides a means for the optical detection of H2 in the upper atmosphere. In particular, the Ly beta line of hydrogen is nearly degenerate with the (6,0) P1 transition, and absorption in this line produces fluorescence in the v-prime = 6 progression, principally at 1265, 1366, 1462 and 1608 A. Absorption by O2 rapidly attenuates the Ly beta from an overhead sun below 100 km and also significantly attenuates the fluorescent radiation. Far-ultraviolet dayglow spectra from 1130 to 1510 A obtained from an Aerobee rocket experiment on 11 December 1972 give an upper limit for any H2 emission which is a factor of 5 higher than expected according to recent hydrogen models.

  14. How to measure separation and angles between inter-molecular fluorescent markers

    DEFF Research Database (Denmark)

    Flyvbjerg, Henrik

    Structure and function of an individual biomolecule can be explored with minimum two fluorescent markers of different colors. Since the light of such markers can be spec- trally separated and imaged simultaneously, the markers can be colocalized. Here, we describe the method used for such two......-color colocalization microscopy. Then we extend it to fluorescent markers with fixed orientations and in intramolecular proximity. Our benchmarking of this extension produced two extra results: (a) we established short double-labeled DNA molecules as probes of 3D orientation of anything to which one can attach them...

  15. Multiplex competitive microbead-based flow cytometric immunoassay using quantum dot fluorescent labels

    International Nuclear Information System (INIS)

    Yu, Hye-Weon; Kim, In S.; Niessner, Reinhard; Knopp, Dietmar

    2012-01-01

    Highlights: ► First time, duplex competitive bead-based flow cytometric immunoassay was developed using ODs. ► Antibody-coated QD detection probes and antigen-immobilized microspheres were synthesized. ► The two model target analytes were low molecular weight compounds of microbial and chemical origin. ► The determination of different water types was possible after simple filtration of samples. - Abstract: In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose–response curves with IC 50 values of 5 μg L −1 and 1.1 μg L −1 and dynamic ranges of 0.52–30 μg L −1 and 0.13–10 μg L −1 were obtained, respectively. Recovery was 92.6–106.5% for 5 types of water samples like bottled

  16. Fluorescence of reduced charge montmorillonite complexes with methylene blue: Experiments and molecular modeling

    Czech Academy of Sciences Publication Activity Database

    Klika, Z.; Pustková, P.; Praus, P.; Kovář, P.; Pospíšil, M.; Malý, P.; Grygar, Tomáš; Kulhánková, L.; Čapková, P.

    2009-01-01

    Roč. 339, č. 2 (2009), s. 416-423 ISSN 0021-9797 Institutional research plan: CEZ:AV0Z40320502 Keywords : methylene blue * reduced charge montmorillonites * fluorescence Subject RIV: DB - Geology ; Mineralogy Impact factor: 3.019, year: 2009

  17. Towards the molecular bases of polymerase dynamics

    International Nuclear Information System (INIS)

    Chela Flores, J.

    1991-03-01

    One aspect of the strong relationship that is known to exist between the processes of DNA replication and transcription is manifest in the coupling of the rates of movement of the replication fork (r f ) and RNA polymerase (r t ). We address two issues concerning the largely unexplored area of polymerase dynamics: (i) The validity of an approximate kinematic formula linking r f and r t suggested by experiments in which transcription is initiated in some prokaryotes with the antibiotic streptolydigin, and (ii) What are the molecular bases of the kinematic formula? An analysis of the available data suggests possible molecular bases for polymerase dynamics. In particular, we are led to a hypothesis: In active chromatin r t may depend on the length (λ t ) of the transcript of the primary messenger RNA (pre-mRNA). This new effect is subject to experimental verification. We discuss possible experiments that may be performed in order to test this prediction. (author). Refs, 6 tabs

  18. Adaptation of Tri-molecular fluorescence complementation allows assaying of regulatory Csr RNA-protein interactions in bacteria.

    Science.gov (United States)

    Gelderman, Grant; Sivakumar, Anusha; Lipp, Sarah; Contreras, Lydia

    2015-02-01

    sRNAs play a significant role in controlling and regulating cellular metabolism. One of the more interesting aspects of certain sRNAs is their ability to make global changes in the cell by interacting with regulatory proteins. In this work, we demonstrate the use of an in vivo Tri-molecular Fluorescence Complementation assay to detect and visualize the central regulatory sRNA-protein interaction of the Carbon Storage Regulatory system in E. coli. The Carbon Storage Regulator consists primarily of an RNA binding protein, CsrA, that alters the activity of mRNA targets and of an sRNA, CsrB, that modulates the activity of CsrA. We describe the construction of a fluorescence complementation system that detects the interactions between CsrB and CsrA. Additionally, we demonstrate that the intensity of the fluorescence of this system is able to detect changes in the affinity of the CsrB-CsrA interaction, as caused by mutations in the protein sequence of CsrA. While previous methods have adopted this technique to study mRNA or RNA localization, this is the first attempt to use this technique to study the sRNA-protein interaction directly in bacteria. This method presents a potentially powerful tool to study complex bacterial RNA protein interactions in vivo. © 2014 Wiley Periodicals, Inc.

  19. Fluorescence enhancement of the aflatoxin B{sub 1} by forming inclusion complexes with some cyclodextrins and molecular modeling study

    Energy Technology Data Exchange (ETDEWEB)

    Aghamohammadi, Mohammad [Department of Chemistry, Faculty of Science, Tarbiat Modarres University, P.O. Box 14115-111, Tehran (Iran, Islamic Republic of); Alizadeh, Naader [Department of Chemistry, Faculty of Science, Tarbiat Modarres University, P.O. Box 14115-111, Tehran (Iran, Islamic Republic of)], E-mail: alizaden@modares.ac.ir

    2007-12-15

    The interaction between the aflatoxin B{sub 1} (AFB{sub 1}) and three cyclodextrins, {alpha}-cyclodextrin ({alpha}-CD), {beta}-cyclodextrin ({beta}-CD) and heptakis-2,6-dimethyl-o-{beta}-cyclodextrin (ome-CD), was studied by spectrofluorescence technique. It was found that the inclusion association behavior occurs for the complexes of cyclodextrins with AFB{sub 1}. The fluorescence of AFB{sub 1} is generally enhanced in the complexes with cyclodextrins in aqueous solutions. The inclusion complex constants of the three types of cyclodextrins at different temperatures were evaluated from Benesi-Hildebrand plot and also by non-linear regression analysis. These cyclodextrins can only form the 1:1 (host:guest) inclusion complex in the studied temperature range of 20-50 deg. C. The enthalpy ({delta}H{sup o}) and entropy ({delta}S{sup o}) changes of complexation were extracted from the temperature dependency of complex formation constants (K). Temperature-dependent measurements showed that the association step is controlled by enthalpy-entropy compensation effect. The use of ome-CD generally resulted in the greatest fluorescence intensity. On the other hand, the discrepancy between the exhibited enhanced fluorescence and thermodynamic parameters ({delta}G{sup o}) is proposed to be different only by the orientation of the AFB{sub 1} within the cyclodextrin cavity. To find the most favorable structure, the geometry of complex was investigated by molecular modeling approach employing the semiemperical HF-SCF calculations.

  20. Fluorescence imaging of the lymph node uptake of proteins in mice after subcutaneous injection: molecular weight dependence.

    Science.gov (United States)

    Wu, Fang; Bhansali, Suraj G; Law, Wing Cheung; Bergey, Earl J; Prasad, Paras N; Morris, Marilyn E

    2012-07-01

    To use noninvasive fluorescence imaging to investigate the influence of molecular weight (MW) of proteins on the rate of loss from a subcutaneous (SC) injection site and subsequent uptake by the draining lymph nodes in mice. Bevacizumab (149 kDa), bovine serum albumin (BSA, 66 kDa), ovalbumin (44.3 kDa) or VEGF-C156S (23 kDa), labeled with the near infrared dye IRDye 680, were injected SC into the front footpad of SKH-1 mice. Whole body non-invasive fluorescence imaging was performed to quantitate the fluorescence signal at the injection site and in axillary lymph nodes. The half-life values, describing the times for 50% loss of proteins from the injection site, were 6.81 h for bevacizumab, 2.85 h for BSA, 1.57 h for ovalbumin and 0.31 h for VEGF-C156S. The corresponding axillary lymph node exposure, represented as the area of the % dose versus time curve, was 6.27, 5.13, 4.06 and 1.54% dose ∙ h, respectively. Our results indicate that the rate of loss of proteins from a SC injection site is inversely related to MW of proteins, while lymph node exposure is proportionally related to the MW of proteins in a mouse model.

  1. Investigation of three flavonoids binding to bovine serum albumin using molecular fluorescence technique

    International Nuclear Information System (INIS)

    Bi Shuyun; Yan Lili; Pang Bo; Wang Yu

    2012-01-01

    The three flavonoids including naringenin, hesperetin and apigenin binding to bovine serum albumin (BSA) at pH 7.4 was studied by fluorescence quenching, synchronous fluorescence and UV-vis absorption spectroscopic techniques. The results obtained revealed that naringenin, hesperetin and apigenin strongly quenched the intrinsic fluorescence of BSA. The Stern-Volmer curves suggested that these quenching processes were all static quenching processes. At 291 K, the value and the order of the binding constant were K A n aringenin) =4.08x10 4 A(hesperetin) =5.40x10 4 ∼K A(apigenin) =5.32x10 4 L mol -1 . The main binding force between the flavonoid and BSA was hydrophobic and electrostatic force. According to the Foerster theory of non-radiation energy transfer, the binding distances (r 0 ) were obtained as 3.36, 3.47 and 3.30 nm for naringenin-BSA, hesperetin-BSA and apigenin-BSA, respectively. The effect of some common ions such as Fe 3+ , Cu 2+ , Mg 2+ , Mn 2+ , Zn 2+ and Ca 2+ on the binding was also studied in detail. The competition binding was also performed. The apparent binding constant (K' A ) obtained suggested that one flavonoid had an obvious effect on the binding of another flavonoid to protein when they coexisted in BSA solution. - Highlights: → Quenchings of BSA fluorescence by the flavonoids was all static quenchings. → Synchronous fluorescence was applied to study the structural change of BSA. → Binding constant, binding site and binding force were determined. → Competition binding experiments were performed. → One flavonoid had an obvious effect on the binding of another one to BSA.

  2. Fluorescent Protein Approaches in Alpha Herpesvirus Research

    Directory of Open Access Journals (Sweden)

    Ian B. Hogue

    2015-11-01

    Full Text Available In the nearly two decades since the popularization of green fluorescent protein (GFP, fluorescent protein-based methodologies have revolutionized molecular and cell biology, allowing us to literally see biological processes as never before. Naturally, this revolution has extended to virology in general, and to the study of alpha herpesviruses in particular. In this review, we provide a compendium of reported fluorescent protein fusions to herpes simplex virus 1 (HSV-1 and pseudorabies virus (PRV structural proteins, discuss the underappreciated challenges of fluorescent protein-based approaches in the context of a replicating virus, and describe general strategies and best practices for creating new fluorescent fusions. We compare fluorescent protein methods to alternative approaches, and review two instructive examples of the caveats associated with fluorescent protein fusions, including describing several improved fluorescent capsid fusions in PRV. Finally, we present our future perspectives on the types of powerful experiments these tools now offer.

  3. Energetics and dynamics of the non-natural fluorescent 4AP:DAP base pair

    KAUST Repository

    Chawla, Mohit

    2018-01-02

    The fluorescent non-natural 4-aminophthalimide (4AP) base, when paired to the complementary 2,4-diaminopyrimidine (DAP) nucleobase, is accommodated in a B-DNA duplex being efficiently recognized and incorporated by DNA polymerases. To complement the experimental studies and rationalize the impact of the above non-natural bases on the structure, stability and dynamics of nucleic acid structures, we performed quantum mechanics (QM) calculations along with classical molecular dynamics (MD) simulations. QM calculations were initially focused on the geometry and energetics of the 4AP:DAP non-natural pair and of H-bonded base pairs between 4AP and all the natural bases in their classical Watson-Crick geometries. The QM calculations indicate that the 4AP:DAP pair, despite the fact that it can form 3 H-bonds in a classic Watson-Crick geometry, has a stability comparable to the A:T pair. Then, we extended the study to reverse Watson-Crick geometries, characteristic of parallel strands. MD simulations were carried out on two 13-mer DNA duplexes, featuring a central 4AP:DAP or A:T pair, respectively. No major structural deformation of the duplex was observed during the MD simulation. Snapshots from the MD simulations were subjected to QM calculations to investigate the 4AP:DAP interaction energy when embedded into a duplex structure, and to investigate the impact of the two non-natural bases on the stacking interactions with adjacent bases in the DNA duplex. We found a slight increase in stacking interactions involving the 4AP:DAP pair, counterbalanced by a moderate decrease in H-bonding interactions of the 4AP:DAP and of the adjacent base pairs in the duplex. The results of our study are in agreement with experimental data and complement them by providing an insight into which factors contribute positively and which factors contribute negatively to the structural compatibility of the fluorescent 4AP:DAP pair with a B-DNA structure.

  4. The use of molecular fluorescent markers to monitor absorption and distribution of xenobiotics in a silkworm model.

    Science.gov (United States)

    Tansil, Natalia C; Li, Yang; Koh, Leng Duei; Peng, Teng Choon; Win, Khin Yin; Liu, Xiang Yang; Han, Ming-Yong

    2011-12-01

    The fate of xenobiotics in living organisms is determined by their in vivo absorption, distribution, metabolism and excretion. A convenient and scalable animal model of these biological processes is thus highly beneficial in understanding the effects of xenobiotics. Here we present a silkworm model to investigate the molecular properties-directed absorption, distribution and excretion of fluorescent compounds as model xenobiotics through introducing the compounds into the silkworm's diet and monitoring the resulting color and fluorescence in the silkworm's body. The efficient uptake of xenobiotics into silk has been further studied through quantitative analysis of the intrinsically colored and highly luminescent silk secreted by silkworm. Our findings provide first-hand insights to better understand the molecular properties that allow specific materials to be incorporated into silk while it is being produced in the silk gland. The use of resulting luminescent silk as scaffold for tissue engineering application has been demonstrated to clearly reveal the interaction of silk with cells. Furthermore, this new development also paves a way to produce various functional silk embedded with stimuli-sensitive dyes or drugs as novel biomaterials for in vivo applications. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. A New Suite of Plasmid Vectors for Fluorescence-Based Imaging of Root Colonizing Pseudomonads

    Directory of Open Access Journals (Sweden)

    Rosemarie Wilton

    2018-02-01

    Full Text Available In the terrestrial ecosystem, plant–microbe symbiotic associations are ecologically and economically important processes. To better understand these associations at structural and functional levels, different molecular and biochemical tools are applied. In this study, we have constructed a suite of vectors that incorporates several new elements into the rhizosphere stable, broad-host vector pME6031. The new vectors are useful for studies requiring multi-color tagging and visualization of plant-associated, Gram-negative bacterial strains such as Pseudomonas plant growth promotion and biocontrol strains. A number of genetic elements, including constitutive promoters and signal peptides that target secretion to the periplasm, have been evaluated. Several next generation fluorescent proteins, namely mTurquoise2, mNeonGreen, mRuby2, DsRed-Express2 and E2-Crimson have been incorporated into the vectors for whole cell labeling or protein tagging. Secretion of mTurquoise2 and mNeonGreen into the periplasm of Pseudomonas fluorescens SBW25 has also been demonstrated, providing a vehicle for tagging proteins in the periplasmic compartment. A higher copy number version of select plasmids has been produced by introduction of a previously described repA mutation, affording an increase in protein expression levels. The utility of these plasmids for fluorescence-based imaging is demonstrated by root colonization of Solanum lycopersicum seedlings by P. fluorescens SBW25 in a hydroponic growth system. The plasmids are stably maintained during root colonization in the absence of selective pressure for more than 2 weeks.

  6. [A cell-based detection of ciguatoxin using sodium fluorescence probe].

    Science.gov (United States)

    Yuan, Jian-hui; Yang, Hui; Tang, Huan-wen; Huang, Wei; Xu, Xin-yun; Liu, Jian-jun; Ke, Yue-bin; Cheng, Jin-quan; Zhuang, Zhi-xiong

    2011-04-01

    To establish a cell-based detection method of ciguatoxin using fluorescence assay. Mouse neuroblastoma N-2A cells were exposed to ouabain and veratridine and different concentrations of standard ciguatoxin samples (P-CTX-1) to establish the curvilinear relationship between the toxin dosage and fluorescence intensity using the sodium fluorescence probe CoroNaTM Green. The toxicity curvilinear relationship was also generated between the toxin dosage and cell survival using CCK-8 method. Based on these standard curves, the presence of ciguatoxin was detected in 33 samples of deep-sea coral fish. A correlation was found between the detection results of cell-based fluorescence assay and cytotoxicity assay, whose detection limit reached 103 g/ml and 1012 g/ml, respectively. The cell-based fluorescent assay sensitivity showed a higher sensitivity than cytotoxicity assay with a 2-4 h reduction of the detection time. The cell-based fluorescent assay can quickly and sensitively detect ciguatoxin and may serve as a good option for preliminary screening of the toxin.

  7. A ratiometric nanoprobe based on silver nanoclusters and carbon dots for the fluorescent detection of biothiols

    Science.gov (United States)

    Zhang, Shuming; Lin, Bixia; Yu, Ying; Cao, Yujuan; Guo, Manli; Shui, Lingling

    2018-04-01

    Ratiometric fluorescent probes could eliminate the influence from experimental factors and improve the detection accuracy. In this article, a ratiometric nanoprobe was constructed based on silver nanoclusters (AgNCs) with nitrogen-doped carbon dots (NCDs) and used for the detection of biothiols. The fluorescence peak of AgNCs was observed at 650 nm with excitation wavelength at 370 nm. In order to construct the ratiometric fluorescent probe, NCDs with the excitation and emission wavelengths at 370 nm and 450 nm were selected. After adding AgNCs, the fluorescence of NCDs was quenched. The mechanism of the fluorescence quenching was studied by fluorescence, UV-Vis absorption and the fluorescence lifetime spectra. The results indicated that the quenching could be ascribed to the inner filter effect (IFE). With the addition of biothiols, the fluorescence of AgNCs at 650 nm decreased due to the breakdown of AgNCs, and the fluorescence of NCDs at 450 nm recovered accordingly. Thus, the relationship between the ratio of the fluorescence intensities (I450/I650) and biothiol concentration was used to establish the determination method for biothiols. Cysteine (Cys) was taken as the model of biothiols, and the working curve for Cys was I450/I650 = 0.60CCys - 1.86 (CCys: μmol/L) with the detection limit of 0.14 μmol/L (S/N = 3). Then, the method was used for the detection of Cys in human urine and serum samples with satisfactory accuracy and recovery ratios. Furthermore, the probe could be applied for the visual semi-quantitative determination of Cys by naked eyes.

  8. Molecular fluorescence enhancement in plasmonic environments: exploring the role of nonlocal effects

    DEFF Research Database (Denmark)

    Tserkezis, Christos; Stefanou, Nikolaos; Wubs, Martijn

    2016-01-01

    strongly affected. In this respect, experimental measurements of fluorescence, the theoretical description of which requires a precise concurrent evaluation of far- and near-field properties of the system, constitute a novel, more sensitive probe for assessing the validity of state-of-the-art nonclassical...... in the nanoparticle vicinity. Here we explore the influence of hitherto disregarded nonclassical effects in the description of emitter-plasmon hybrids, focusing on the roles of metal nonlocal response and especially size-dependent plasmon damping. Through extensive modelling of metallic nanospheres and nanoshells...... coupled to dipole emitters, we show that within a purely classical description a remarkable fluorescence enhancement can be achieved. However, once departing from the local-response approximation, and particularly by implementing the recent generalised nonlocal optical response theory, which provides...

  9. A novel molecular Fluorescent technique for Imaging the Somatostatin Receptor 2 Using a DOTATOC Lanthanide Conjugate

    DEFF Research Database (Denmark)

    Andersen, Rune Wiik; Prakash, Vineet; Stensballe, Allan

    for synaptophysin.                         RESULTS            It is feasible to usefully chelate Samarium and Europium to DOTATOC. There is a distinct higher fluorescent signal arising from the chelation of the two ions than by the DOTA functional group alone. The unparaffinated pancreatic tumor tissues demonstrate.......                       CLINICAL RELEVANCE/APPLICATION            We propose a method for the histopatholgical receptor verification using fluorescent DOTATOC imaging. This potentially permits  ex-vivo developmental platforms for DOTA-conjugated molecules.        ...

  10. Oxidation Changes Physical Properties of Phospholipid Bilayers: Fluorescence Spectroscopy and Molecular Simulations

    Czech Academy of Sciences Publication Activity Database

    Beranová, Lenka; Cwiklik, Lukasz; Jurkiewicz, Piotr; Hof, Martin; Jungwirth, Pavel

    2010-01-01

    Roč. 26, č. 9 (2010), s. 6140-6144 ISSN 0743-7463 R&D Projects: GA MŠk LC512; GA MŠk(CZ) LC06063; GA AV ČR GEMEM/09/E006 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z40550506 Keywords : oxidized phospholipides * membranes * fluorescence spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 4.269, year: 2010

  11. Thioflavin T as a fluorescence probe for monitoring RNA metabolism at molecular and cellular levels

    OpenAIRE

    Sugimoto, Shinya; Arita-Morioka, Ken-ichi; Mizunoe, Yoshimitsu; Yamanaka, Kunitoshi; Ogura, Teru

    2015-01-01

    The intrinsically stochastic dynamics of mRNA metabolism have important consequences on gene regulation and non-genetic cell-to-cell variability; however, no generally applicable methods exist for studying such stochastic processes quantitatively. Here, we describe the use of the amyloid-binding probe Thioflavin T (ThT) for monitoring RNA metabolism in vitro and in vivo. ThT fluoresced strongly in complex with bacterial total RNA than with genomic DNA. ThT bound purine oligoribonucleotides pr...

  12. Lanthanide-based fluorescent tracers in complex media

    International Nuclear Information System (INIS)

    Brichart, Thomas

    2014-01-01

    Tracers are objects allowing the determination of the position or the distribution of a product; tracers are currently used in a great variety of domains. Despite the fact that each field has it's own specifications, it is possible to find tracers in medicine (contrast agents), anti-counterfeiting or geological exploration. We have developed lanthanide complex tracers for oil field injection waters. Those tracers, derived from the DOTA, have been detected at concentration lower than 1 ppb, thanks to a simple and compact apparatus. This detection has been made possible by the use of time-resolved fluorescence spectroscopy, this technique allows us to get rid of the background noise created by the intrinsic fluorescence of oil residues that are present in production waters. We also demonstrated how we can, through a reverse microemulsion synthesis, encapsulate several different dyes inside a single nanoparticle composed of a gold core and a silica shell. We showed as well, how those particles can be used as smart tracers to gather data, such as temperature, pH, solvents, etc. inside the well. Finally the use of lanthanides and scale inhibitors properties allowed us to create a simple and fast dosing protocol of such scale inhibitors in injection waters. This dosage will then allow the quick adjustment of their concentration inside each well. (author) [fr

  13. Investigation on the interactions of clenbuterol to bovine serum albumin and lysozyme by molecular fluorescence technique.

    Science.gov (United States)

    Bi, Shuyun; Pang, Bo; Wang, Tianjiao; Zhao, Tingting; Yu, Wang

    2014-01-01

    Clenbuterol interacting with bovine serum albumin (BSA) or lysozyme (LYS) in physiological buffer (pH 7.4) was investigated by the fluorescence spectroscopy and UV-vis absorption spectroscopy. The results indicated that clenbuterol quenched the intrinsic fluorescence of BSA and LYS via a static quenching procedure. The binding constants of clenbuterol with BSA and LYS were 1.16×10(3) and 1.49×10(3) L mol(-1) at 291 K. The values of ΔH and ΔS implied that hydrophobic and electrostatic interaction played a major role in stabilizing the complex (clenbuterol-BSA or clenbuterol-LYS). In the presence of Fe2+, Fe3+, Cu2+, Mg2+, Ca2+, or Zn2+, the binding constants of clenbuterol to BSA or LYS had no significant differences. The distances between the donor (BSA or LYS) and acceptor (clenbuterol) were 2.61 and 2.19 nm for clenbuterol-BSA and clenbuterol-LYS respectively. Furthermore, synchronous fluorescence spectrometry was used to analyze the conformational changes of BSA and LYS. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Exploring viscosity, polarity and temperature sensitivity of BODIPY-based molecular rotors.

    Science.gov (United States)

    Vyšniauskas, Aurimas; López-Duarte, Ismael; Duchemin, Nicolas; Vu, Thanh-Truc; Wu, Yilei; Budynina, Ekaterina M; Volkova, Yulia A; Peña Cabrera, Eduardo; Ramírez-Ornelas, Diana E; Kuimova, Marina K

    2017-09-27

    Microviscosity is a key parameter controlling the rate of diffusion and reactions on the microscale. One of the most convenient tools for measuring microviscosity is by fluorescent viscosity sensors termed 'molecular rotors'. BODIPY-based molecular rotors in particular proved extremely useful in combination with fluorescence lifetime imaging microscopy, for providing quantitative viscosity maps of living cells as well as measuring dynamic changes in viscosity over time. In this work, we investigate several new BODIPY-based molecular rotors with the aim of improving on the current viscosity sensing capabilities and understanding how the structure of the fluorophore is related to its function. We demonstrate that due to subtle structural changes, BODIPY-based molecular rotors may become sensitive to temperature and polarity of their environment, as well as to viscosity, and provide a photophysical model explaining the nature of this sensitivity. Our data suggests that a thorough understanding of the photophysics of any new molecular rotor, in environments of different viscosity, temperature and polarity, is a must before moving on to applications in viscosity sensing.

  15. A coumarin-quinolinium-based fluorescent probe for ratiometric sensing of sulfite in living cells.

    Science.gov (United States)

    Tan, Li; Lin, Weiying; Zhu, Sasa; Yuan, Lin; Zheng, Kaibo

    2014-07-14

    Based on a novel coumarin-quinolinium platform, probe 2 was rationally designed and synthesized as a novel ratiometric fluorescent sensor for sulfite anions. The probe exhibited a wide dynamic concentration range for sulfite anions in a PBS buffer (containing 1 mg mL(-1) BSA). More importantly, the probe was suitable for ratiometric fluorescence imaging in living cells with high sensitivity, favorable selectivity, and minimal cytotoxicity.

  16. Combined quantum-mechanical molecular mechanics calculations with NWChem and AMBER: Excited state properties of green fluorescent protein chromophore analogue in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Pirojsirikul, Teerapong [Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive La Jolla California 92093; Götz, Andreas W. [San Diego Supercomputer Center, University of California San Diego, 9500 Gilman Drive La Jolla California 92093; Weare, John [Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive La Jolla California 92093; Walker, Ross C. [Department of Chemistry and Biochemistry, University of California San Diego, 9500 Gilman Drive La Jolla California 92093; GlaxoSmithKline, 1250 S. Collegeville Road Collegeville Pennsylvania 19426; Kowalski, Karol [Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P. O. Box 999 Richland Washington 99352; Valiev, Marat [Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P. O. Box 999 Richland Washington 99352

    2017-05-03

    Green Fluorescent Protein (GFP) is a widely used fluorescent biomarker for the study of biological systems. Our investigation is focused on providing a reliable theoretical description of the GFP chromophore, the photochemical properties of which can be influenced through both the surrounding protein environment and pH levels. In this work we are specifically addressing the effect of an aqueous solvation environment , where a number of experimental measurements have been performed. Our approach is based on a combined quantum mechanics molecular mechanics (QM/MM) methodology, which incorporates high level coupled cluster theory for the analysis of excited states. It also presents the first application of the newly developed NWChem/AMBER QM/MM interface. Using a systematic approach, which involves comparison of gas phase and aqueous results for different protonation states and conformations, we have resolved existing uncertainties regarding theoretical interpretation of the experimental data. We observe that the impact of aqueous environment on charged states generally results in blue shifts, but the magnitude of the effect is sensitive to charge state and conformation and can be rationalized based on charge movement into the area of higher/lower external electrostatic potentials. At neutral pH levels the experimentally observed absorption signal is most likely coming from the phenol protonated form. Our results also show that the high level coupled description is essential for proper description of excited states of GFP.

  17. Homogeneous non-competitive bioaffinity assay based on fluorescence resonance energy transfer

    International Nuclear Information System (INIS)

    Kokko, Tiina; Kokko, Leena; Soukka, Tero; Loevgren, Timo

    2007-01-01

    A homogeneous non-competitive assay principle for measurement of small analytes based on quenching of fluorescence is described. Fluorescence resonance energy transfer (FRET) occurs between the donor, intrinsically fluorescent europium(III)-chelate conjugated to streptavidin, and the acceptor, quencher dye conjugated to biotin derivative when the biotin-quencher is bound to Eu-streptavidin. Fluorescence can be measured only from those streptavidins that are bound to biotin of the sample, while the fluorescence of the streptavidins that are not occupied by biotin are quenched by quencher-biotin conjugates. The quenching efficiencies of the non-fluorescent quencher dyes were over 95% and one dye molecule was able to quench the fluorescence of more than one europium(III)-chelate. This, however, together with the quadrovalent nature of streptavidin limited the measurable range of the assay to 0.2-2 nmol L -1 . In this study we demonstrated that FRET could be used to design a non-competitive homogeneous assay for a small analyte resulting in equal performance with competitive heterogeneous assay

  18. Oral cancer detection based on fluorescence polarization of blood plasma at excitation wavelength 405 nm

    Science.gov (United States)

    Pachaiappan, Rekha; Prakasarao, Aruna; Manoharan, Yuvaraj; Dornadula, Koteeswaran; Singaravelu, Ganesan

    2017-02-01

    During metabolism the metabolites such as hormones, proteins and enzymes were released in to the blood stream by the cells. These metabolites reflect any change that occurs due to any disturbances in normal metabolic function of the human system. This was well observed with the altered spectral signatures observed with fluorescence spectroscopic technique. Previously many have reported on the significance of native fluorescence spectroscopic method in the diagnosis of cancer. As fluorescence spectroscopy is sensitive and simple, it has complementary techniques such as excitation-emission matrix, synchronous and polarization. The fluorescence polarization measurement provides details about any association or binding reactions and denaturing effects that occurs due to change in the micro environment of cells and tissues. In this study, we have made an attempt in the diagnosis of oral cancer at 405 nm excitation using fluorescence polarization measurement. The fluorescence anisotropic values calculated from polarized fluorescence spectral data of normal and oral cancer subjects yielded a good accuracy when analyzed with linear discriminant analysis based artificial neural network. The results will be discussed in detail.

  19. A new Schiff base based on vanillin and naphthalimide as a fluorescent probe for Ag+ in aqueous solution

    Science.gov (United States)

    Zhou, Yanmei; Zhou, Hua; Ma, Tongsen; Zhang, Junli; Niu, Jingyang

    2012-03-01

    A new Schiff base based on vanillin and naphthalimide was designed and synthesized as fluorescent probe. The probe showed high selectivity for Ag+ over other metal ions such as Pb2+, Na+, K+, Cd2+, Ba2+, Cr3+, Zn2+, Cu2+, Ni2+, Ca2+, Al3+ and Mg2+ in aqueous solution. A new fluorescence emission was observed at 682 nm in the presence of Ag+ ion. The fluorescence intensity quenched with increasing the concentration of Ag+ at 682 nm. The method of job's plot confirmed the 1:2 complex between Ag+ and probe, and the mechanism was proposed.

  20. Feasibility of Raman spectroscopy in vitro after 5-ALA-based fluorescence diagnosis in the bladder

    Science.gov (United States)

    Grimbergen, M. C. M.; van Swol, C. F. P.; van Moorselaar, R. J. A.; Mahadevan-Jansen, A.,; Stone, N.

    2006-02-01

    Photodynamic diagnosis (PDD) has become popular in bladder cancer detection. Several studies have however shown an increased false positive biopsies rate under PDD guidance compared to conventional cystoscopy. Raman spectroscopy is an optical technique that utilizes molecular specific, inelastic scattering of light photons to interrogate biological tissues, which can successfully differentiate epithelial neoplasia from normal tissue and inflammations in vitro. This investigation was performed to show the feasibility of NIR Raman spectroscopy in vitro on biopsies obtained under guidance of 5-ALA induced PPIX fluorescence imaging. Raman spectra of a PPIX solution was measured to obtain a characteristic signature for the photosensitzer without contributions from tissue constituents. Biopsies were obtained from patients with known bladder cancer instilled with 50ml, 5mg 5-ALA two hours prior to trans-urethral resection of tumor (TURT). Additional biopsies were obtained at a fluorescent and non-fluorescent area, snap-frozen in liquid nitrogen and stored at -80 °C. Each biopsy was thawed before measurements (10sec integration time) with a confocal Raman system (Renishaw Gloucestershire, UK). The 830 nm excitation (300mW) source is focused on the tissue by a 20X ultra-long-working-distance objective. Differences in fluorescence background between the two groups were removed by means of a special developed fluorescence subtraction algorithm. Raman spectra from ALA biopsies showed different fluorescence background which can be effectively removed by a fluorescence subtraction algorithm. This investigation shows that the interaction of the ALA induced PPIX with Raman spectroscopy in bladder samples. Combination of these techniques in-vivo may lead to a viable method of optical biopsies in bladder cancer detection.

  1. Fluorescence-based sorting of neural stem cells and progenitors.

    Science.gov (United States)

    Maric, Dragan; Barker, Jeffery L

    2005-11-01

    Neural stem cells (NSCs) are defined as undifferentiated cells originating from the neuroectoderm that have the capacity both to perpetually self-renew without differentiating and to generate multiple types of lineage-restricted progenitors (LRPs). LRPs can themselves undergo limited self-renewal and ultimately differentiate into highly specialized cells that make up the nervous system. However, this physiologically delimited definition of NSCs and LRPs has become increasingly blurred due to lack of protocols for effectively separating these types of cells from primary tissues. This unit discusses recent attempts using fluorescence-activated cell sorting (FACS) strategies to prospectively isolate NSCs from different types of LRPs as they appear in vivo, and details a protocol that optimally attains this goal. Thus, the strategy presented here provides a framework for more precise studies of NSC and LRP cell biology in the future, which can be applied to all vertebrates, including humans.

  2. Molecularly Imprinted Core-Shell CdSe@SiO2/CDs as a Ratiometric Fluorescent Probe for 4-Nitrophenol Sensing

    Science.gov (United States)

    Liu, Mingyue; Gao, Zhao; Yu, Yanjun; Su, Rongxin; Huang, Renliang; Qi, Wei; He, Zhimin

    2018-01-01

    4-Nitrophenol (4-NP) is a priority pollutant in water and is both carcinogenic and genotoxic to humans and wildlife even at very low concentrations. Thus, we herein fabricated a novel molecularly imprinted core-shell nanohybrid as a ratiometric fluorescent sensor for the highly sensitive and selective detection of 4-NP. This sensor was functioned by the transfer of fluorescence resonance energy between photoluminescent carbon dots (CDs) and 4-NP. This sensor was synthesized by linking organosilane-functionalized CDs to silica-coated CdSe quantum dots (CdSe@SiO2) via Si-O bonds. The nanohybrids were further modified by anchoring a molecularly imprinted polymer (MIP) layer on the ratiometric fluorescent sensor through a facile sol-gel polymerization method. The morphology, chemical structure, and optical properties of the resulting molecularly imprinted dual-emission fluorescent probe were characterized by transmission electron microscopy and spectroscopic analysis. The probe was then applied in the detection of 4-NP and exhibited good linearity between 0.051 and 13.7 μg/mL, in addition to a low detection limit of 0.026 μg/mL. Furthermore, the simplicity, reliability, high selectivity, and high sensitivity of the developed sensor demonstrate that the combination of MIPs and ratiometric fluorescence allows the preparation of excellent fluorescent sensors for the detection of trace or ultra-trace analytes.

  3. Tuning Selectivity of Fluorescent Carbon Nanotube-Based Neurotransmitter Sensors.

    Science.gov (United States)

    Mann, Florian A; Herrmann, Niklas; Meyer, Daniel; Kruss, Sebastian

    2017-06-28

    Detection of neurotransmitters is an analytical challenge and essential to understand neuronal networks in the brain and associated diseases. However, most methods do not provide sufficient spatial, temporal, or chemical resolution. Near-infrared (NIR) fluorescent single-walled carbon nanotubes (SWCNTs) have been used as building blocks for sensors/probes that detect catecholamine neurotransmitters, including dopamine. This approach provides a high spatial and temporal resolution, but it is not understood if these sensors are able to distinguish dopamine from similar catecholamine neurotransmitters, such as epinephrine or norepinephrine. In this work, the organic phase (DNA sequence) around SWCNTs was varied to create sensors with different selectivity and sensitivity for catecholamine neurotransmitters. Most DNA-functionalized SWCNTs responded to catecholamine neurotransmitters, but both dissociation constants ( K d ) and limits of detection were highly dependent on functionalization (sequence). K d values span a range of 2.3 nM (SWCNT-(GC) 15 + norepinephrine) to 9.4 μM (SWCNT-(AT) 15 + dopamine) and limits of detection are mostly in the single-digit nM regime. Additionally, sensors of different SWCNT chirality show different fluorescence increases. Moreover, certain sensors (e.g., SWCNT-(GT) 10 ) distinguish between different catecholamines, such as dopamine and norepinephrine at low concentrations (50 nM). These results show that SWCNTs functionalized with certain DNA sequences are able to discriminate between catecholamine neurotransmitters or to detect them in the presence of interfering substances of similar structure. Such sensors will be useful to measure and study neurotransmitter signaling in complex biological settings.

  4. Fluorescence ELISA for sensitive detection of ochratoxin A based on glucose oxidase-mediated fluorescence quenching of CdTe QDs

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Yi [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047 (China); Huang, Xiaolin [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Yu, Ruijin [College of Science, Northwest A& F University, Yangling, Shaanxi 712100 (China); Zhou, Yaofeng [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Xiong, Yonghua, E-mail: yhxiongchen@163.com [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047 (China)

    2016-09-14

    The present study described a novel fluorescence enzyme-linked immunosorbent assay (ELISA) used to detect ochratoxin A (OTA) by using the glucose oxidase (GOx)-mediated fluorescence quenching of mercaptopropionic acid-capped CdTe quantum dots (MPA-QDs), in which GOx was used as an alternative to horseradish peroxidase (HRP) for the oxidization of glucose into hydrogen peroxide (H{sub 2}O{sub 2}) and gluconic acid. The MPA-QDs were used as a fluorescent signal output, whose fluorescence variation was extremely sensitive to the presence of H{sub 2}O{sub 2} or hydrogen ions in the solution. Under the optimized conditions, the proposed fluorescence ELISA demonstrated a good linear detection of OTA in corn extract from 2.4 pg mL{sup −1} to 625 pg mL{sup −1} with a limit of detection of 2.2 pg mL{sup −1}, which was approximately 15-fold lower than that of conventional HRP-based ELISA. Our developed fluorescence immunoassay was also similar to HRP-based ELISA in terms of selectivity, accuracy, and reproducibility. In summary, this study was the first to use the GOx-mediated fluorescence quenching of QDs in immunoassay to detect OTA, offering a new possibility for the analysis of other mycotoxins and biomolecules. - Highlights: • A novel fluorescence ELISA was first developed for the detection of OTA by using GOx-mediated fluorescence quenching of QDs. • The pH- and H{sub 2}O{sub 2}-sensitive MPA-capped CdTe QDs were used as a fluorescent signal output to improve the detection sensitivity. • This novel method open up a different vision to detect other mycotoxins and biomolecules.

  5. Use of fluorescence spectroscopy to measure molecular autofluorescence in diabetic subjects

    International Nuclear Information System (INIS)

    Gomes, Cinthia Zanini

    2011-01-01

    Diabetes Mellitus (DM) comprises a complex metabolic syndrome, caused by reduced or absent secretion of insulin by pancreatic beta cells, leading to hyperglycemia. Hyperglycemia promotes glycation of proteins and, consequently, the appearance of advanced glycation end products (AGEs). Currently, diabetic patients are monitored by determining levels of glucose and glycated hemoglobin (HbA1c). The complications caused by hyperglycemia may be divided into micro and macrovascular complications, represented by retinopathy, nephropathy, neuropathy and cardiovascular disease. Albumin (HSA) is the most abundant serum protein in the human body and is subject to glycation. The Protoporphyrin IX (PpIX) is the precursor molecule of heme synthesis, structural component of hemoglobin. The in vitro and animals studies have indicated that hyperglycemia promotes a decrease in its concentration in erythrocytes. The fluorescence spectroscopy is a technique widely used in biomedical field. The autofluorescence corresponds to the intrinsic fluorescence present in some molecules, this being associated with the same structure. The aim of this study was to use fluorescence spectroscopy to measure levels of erythrocyte PpIX autofluorescence and AGE-HSA in diabetic and healthy subjects and compare them with levels of blood glucose and HbA1c. This study was conducted with 151 subjects (58 controls and 93 diabetics). Epidemiological data of patients and controls were obtained from medical records. For control subjects, blood glucose levels were obtained from medical records and levels of Hb1Ac obtained by using commercial kits. The determination of the PpIX autofluorescence was performed with excitation at 405 nm and emission at 632 nm. Determination of AGE-HSA was performed with excitation at 370 nm and emission at 455 nm. Approximately 50% of diabetic had micro and macrovascular lesions resulting from hyperglycemia. There were no significant differences in the PpIX emission intensity values

  6. As bases moleculares da hemofilia A

    Directory of Open Access Journals (Sweden)

    Simone Ferreira Pio

    2009-01-01

    Full Text Available As hemofilias são doenças hemorrágicas resultantes da deficiência de fator VIII (hemofilia A ou de fator IX (hemofilia B da coagulação, decorrentes de mutações nos genes que codificam os fatores VIII ou IX, respectivamente. A hemofilia A é mais frequente que a hemofilia B e acomete aproximadamente 1:10.000 nascimentos masculinos. A gravidade e frequência dos episódios hemorrágicos está relacionado ao nível residual de atividade de fator VIII presente no plasma e este relaciona-se ao tipo de mutação associada à doença. A clonagem do gene do fator VIII tornou possível o conhecimento das bases moleculares da hemofilia A, sendo hoje conhecidas mais de 1.000 mutações associadas à doença. O conhecimento das bases moleculares da hemofilia A permite uma melhor compreensão da relação genótipo-fenótipo da doença, tomada de condutas clínicas diferenciadas em casos de mutações associadas a um maior risco de desenvolvimento de inibidor, determinação da condição de portadora de hemofilia em mulheres relacionadas aos pacientes, implementação de programa de aconselhamento genético/orientação familiar e melhor compreensão das relações estruturais-funcionais do gene-proteína. Este artigo propõe revisar as bases moleculares da hemofilia A, os métodos laboratoriais utilizados para a caracterização das mutações e as implicações clínicas envolvidas no diagnóstico molecular da hemofilia A.Hemophilias are bleeding disorders due to deficiency of the blood coagulation factor VIII (hemophilia A or factor IX (hemophilia B, resulting from mutation on the gene coding for factor VIII or factor IX. Hemophilia A is more frequent than hemophilia B and affects 1:10,000 male newborns. The severity and frequency of hemorrhagic episodes is related to residual activity of factor VIII present in the plasma and relates to the type of mutation associated with the disorder. Cloning of the factor VIII gene has enabled researchers to

  7. Pentamethinium fluorescent probes: The impact of molecular structure on photophysical properties and subcellular localization

    Czech Academy of Sciences Publication Activity Database

    Bříza, T.; Rimpelová, S.; Králová, Jarmila; Záruba, K.; Kejík, Z.; Ruml, T.; Martásek, P.; Král, V.

    2014-01-01

    Roč. 107, August 2014 (2014), s. 51-59 ISSN 0143-7208 R&D Projects: GA AV ČR KAN200100801; GA ČR(CZ) GAP303/11/1291; GA MŠk(CZ) CZ.01.05/2.1.00/01.00.30; GA MŠk(CZ) CZ.1.07/2.300/30.0060 Institutional support: RVO:68378050 Keywords : Pentamethinium salts * Fluorescent probes * Mitochondria * Cardiolipin Photostability * Organelle imaging Subject RIV: CE - Biochemistry Impact factor: 3.966, year: 2014

  8. Pixel-based absorption correction for dual-tracer fluorescence imaging of receptor binding potential.

    Science.gov (United States)

    Kanick, Stephen C; Tichauer, Kenneth M; Gunn, Jason; Samkoe, Kimberley S; Pogue, Brian W

    2014-10-01

    Ratiometric approaches to quantifying molecular concentrations have been used for decades in microscopy, but have rarely been exploited in vivo until recently. One dual-tracer approach can utilize an untargeted reference tracer to account for non-specific uptake of a receptor-targeted tracer, and ultimately estimate receptor binding potential quantitatively. However, interpretation of the relative dynamic distribution kinetics is confounded by differences in local tissue absorption at the wavelengths used for each tracer. This study simulated the influence of absorption on fluorescence emission intensity and depth sensitivity at typical near-infrared fluorophore wavelength bands near 700 and 800 nm in mouse skin in order to correct for these tissue optical differences in signal detection. Changes in blood volume [1-3%] and hemoglobin oxygen saturation [0-100%] were demonstrated to introduce substantial distortions to receptor binding estimates (error > 30%), whereas sampled depth was relatively insensitive to wavelength (error pixel-by-pixel normalization of tracer inputs immediately post-injection was found to account for spatial heterogeneities in local absorption properties. Application of the pixel-based normalization method to an in vivo imaging study demonstrated significant improvement, as compared with a reference tissue normalization approach.

  9. Pixel-based absorption correction for dual-tracer fluorescence imaging of receptor binding potential

    Science.gov (United States)

    Kanick, Stephen C.; Tichauer, Kenneth M.; Gunn, Jason; Samkoe, Kimberley S.; Pogue, Brian W.

    2014-01-01

    Ratiometric approaches to quantifying molecular concentrations have been used for decades in microscopy, but have rarely been exploited in vivo until recently. One dual-tracer approach can utilize an untargeted reference tracer to account for non-specific uptake of a receptor-targeted tracer, and ultimately estimate receptor binding potential quantitatively. However, interpretation of the relative dynamic distribution kinetics is confounded by differences in local tissue absorption at the wavelengths used for each tracer. This study simulated the influence of absorption on fluorescence emission intensity and depth sensitivity at typical near-infrared fluorophore wavelength bands near 700 and 800 nm in mouse skin in order to correct for these tissue optical differences in signal detection. Changes in blood volume [1-3%] and hemoglobin oxygen saturation [0-100%] were demonstrated to introduce substantial distortions to receptor binding estimates (error > 30%), whereas sampled depth was relatively insensitive to wavelength (error tracer inputs immediately post-injection was found to account for spatial heterogeneities in local absorption properties. Application of the pixel-based normalization method to an in vivo imaging study demonstrated significant improvement, as compared with a reference tissue normalization approach. PMID:25360349

  10. An in-situ fluorescence-based optical extensometry system for imaging mechanically loaded bone.

    Science.gov (United States)

    Price, Christopher; Li, Wen; Novotny, John E; Wang, Liyun

    2010-06-01

    The application and quantification of well-controlled tissue strains is required for investigations into mechanisms of tissue adaptation within the musculoskeletal system. Although many commercial and custom extensometry systems exist for large biological samples, integrated loading/strain measurement for small samples is not as readily available. Advanced imaging modules such as laser scanning microscopy provide in situ, minimally invasive tools to probe cellular and molecular processes with high spatiotemporal resolution. Currently, a need exists to devise loading/strain measurement systems that can be integrated with such advanced imaging modules. We describe the development and validation of a fluorescence-based, optical extensometry system directly integrated within a confocal microscopy platform. This system allows in situ measurement of surface strain and is compatible with the direct imaging of cellular processes within small bone samples. This optical extensometry system can accurately and reproducibly measure physiologically relevant surface strains (200 to 3000 microstrain) in beams machined from various well-characterized materials, including bovine femoral cortex, and in intact murine tibia. This simple system provides a powerful tool to further our investigation of the relationships between mechanical loading, fluid and solute transport, and mechanosensation within the musculoskeletal system. (c) 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  11. Eco-friendly carbon-nanodot-based fluorescent paints for advanced photocatalytic systems

    Science.gov (United States)

    Young Park, So; Uk Lee, Hyun; Lee, Young-Chul; Choi, Saehae; Hyun Cho, Dae; Sik Kim, Hee; Bang, Sunghee; Seo, Soonjoo; Chang Lee, Soon; Won, Jonghan; Son, Byung-Chul; Yang, Mino; Lee, Jouhahn

    2015-07-01

    Fluorescent carbon nanomaterials, especially zero-dimensional (0D) carbon nanodots (CDs), are widely used in broad biological and optoelectronic applications. CDs have unique characteristics such as strong fluorescence, biocompatibility, sun-light response, and capability of mass-production. Beyond the previous green CD obtained from harmful natural substances, we report a new type of fluid-based fluorescent CD paints (C-paints) derived from polyethylene glycol (PEG; via simple ultrasound irradiation at room temperatures) and produced in quantum yields of up to ~14%. Additionally, C-paints possess a strong, UV- and visible-light-responsive photoluminescent (PL) property. Most especially, C-paints, by incorporation into a photocatalytic system, show additional roles in the emission of fluorescent light for activation of TiO2 nanoparticles (NPs) and the resultant detoxification of most organic dyes, thus further enabling embarkation in advanced water purification.

  12. Eco-friendly carbon-nanodot-based fluorescent paints for advanced photocatalytic systems.

    Science.gov (United States)

    Park, So Young; Lee, Hyun Uk; Lee, Young-Chul; Choi, Saehae; Cho, Dae Hyun; Kim, Hee Sik; Bang, Sunghee; Seo, Soonjoo; Lee, Soon Chang; Won, Jonghan; Son, Byung-Chul; Yang, Mino; Lee, Jouhahn

    2015-07-23

    Fluorescent carbon nanomaterials, especially zero-dimensional (0D) carbon nanodots (CDs), are widely used in broad biological and optoelectronic applications. CDs have unique characteristics such as strong fluorescence, biocompatibility, sun-light response, and capability of mass-production. Beyond the previous green CD obtained from harmful natural substances, we report a new type of fluid-based fluorescent CD paints (C-paints) derived from polyethylene glycol (PEG; via simple ultrasound irradiation at room temperatures) and produced in quantum yields of up to ~14%. Additionally, C-paints possess a strong, UV- and visible-light-responsive photoluminescent (PL) property. Most especially, C-paints, by incorporation into a photocatalytic system, show additional roles in the emission of fluorescent light for activation of TiO2 nanoparticles (NPs) and the resultant detoxification of most organic dyes, thus further enabling embarkation in advanced water purification.

  13. Determination of flumioxazin residue in food samples through a sensitive fluorescent sensor based on click chemistry.

    Science.gov (United States)

    Lu, Lijun; Yang, Linlin; Cai, Huijian; Zhang, Lan; Lin, Zhenyu; Guo, Longhua; Qiu, Bin; Chen, Guonan

    2014-11-01

    A sensitive and selective fluorescent sensor for flumioxazin was designed based on the formation of strong fluorescence compound (1,2,3-triazole compounds) via the reaction of the alkynyl group in flumioxazin with 3-azido-7-hydroxycoumarin, a weak-fluorescent compound, through the Cu(+)-catalysed azide-alkyne cycloaddition (CuAAC) reaction. The fluorescence increase factor (represented by F/F0) of the system exhibited a good linear relationship with the concentrations of flumioxazin in the range of 0.25-6.0 μg/L with a detection limit of 0.18 μg/L (S/N=3). Also, the proposed fluorescent sensor demonstrated good selectivity for flumioxazin assay even in the presence of high concentration of other pesticides. Based on such high sensitivity and selectivity, the proposed fluorescent sensor has been applied to test the flumioxazin residue in some vegetable and water samples with satisfied results. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Static Hyperspectral Fluorescence Imaging of Viscous Materials Based on a Linear Variable Filter Spectrometer

    Directory of Open Access Journals (Sweden)

    Alexander W. Koch

    2013-09-01

    Full Text Available This paper presents a low-cost hyperspectral measurement setup in a new application based on fluorescence detection in the visible (Vis wavelength range. The aim of the setup is to take hyperspectral fluorescence images of viscous materials. Based on these images, fluorescent and non-fluorescent impurities in the viscous materials can be detected. For the illumination of the measurement object, a narrow-band high-power light-emitting diode (LED with a center wavelength of 370 nm was used. The low-cost acquisition unit for the imaging consists of a linear variable filter (LVF and a complementary metal oxide semiconductor (CMOS 2D sensor array. The translucent wavelength range of the LVF is from 400 nm to 700 nm. For the confirmation of the concept, static measurements of fluorescent viscous materials with a non-fluorescent impurity have been performed and analyzed. With the presented setup, measurement surfaces in the micrometer range can be provided. The measureable minimum particle size of the impurities is in the nanometer range. The recording rate for the measurements depends on the exposure time of the used CMOS 2D sensor array and has been found to be in the microsecond range.

  15. Quantifying the Assembly of Multicomponent Molecular Machines by Single-Molecule Total Internal Reflection Fluorescence Microscopy.

    Science.gov (United States)

    Boehm, E M; Subramanyam, S; Ghoneim, M; Washington, M Todd; Spies, M

    2016-01-01

    Large, dynamic macromolecular complexes play essential roles in many cellular processes. Knowing how the components of these complexes associate with one another and undergo structural rearrangements is critical to understanding how they function. Single-molecule total internal reflection fluorescence (TIRF) microscopy is a powerful approach for addressing these fundamental issues. In this article, we first discuss single-molecule TIRF microscopes and strategies to immobilize and fluorescently label macromolecules. We then review the use of single-molecule TIRF microscopy to study the formation of binary macromolecular complexes using one-color imaging and inhibitors. We conclude with a discussion of the use of TIRF microscopy to examine the formation of higher-order (i.e., ternary) complexes using multicolor setups. The focus throughout this article is on experimental design, controls, data acquisition, and data analysis. We hope that single-molecule TIRF microscopy, which has largely been the province of specialists, will soon become as common in the tool box of biophysicists and biochemists as structural approaches have become today. © 2016 Elsevier Inc. All rights reserved.

  16. Pentavalent antimony uptake pathway through erythrocyte membranes: molecular and atomic fluorescence approaches.

    Science.gov (United States)

    Barrera, Camila; López, Silvana; Aguilar, Luis; Mercado, Luis; Bravo, Manuel; Quiroz, Waldo

    2016-04-01

    Previous studies by our group have shown that Sb(V) is able to enter red blood cells in a dynamic process and is reduced to Sb(III) by glutathione. The present study aims to investigate a possible entry pathway for Sb(V) through the erythrocyte membrane. Applying fluorescence spectroscopy studies with Laurdan and diphenylhexatriene (DPH) probes, it was found that there was no interaction between Sb(V) and membrane lipids. By comparing the Sb(V) entry percentages through lipid vesicles and sealed erythrocyte membranes, it was found that Sb(V) required protein channels to pass through the membrane. The competitive inhibition results using HCO3 (-) and Cl(-) showed that the Sb(V) uptake rate through the membrane fell approximately 50-70 % until full inhibition was reached, which was possibly due to the inhibition of the anion exchanger 1 (AE1) channel. Finally, the fluorescence measurements with the 5-iodoacetamidofluorescein (5-IAF) probe showed that Sb(V) interacted with membrane protein SH groups during this process.

  17. Field portable mobile phone based fluorescence microscopy for detection of Giardia lamblia cysts in water samples

    Science.gov (United States)

    Ceylan Koydemir, Hatice; Gorocs, Zoltan; McLeod, Euan; Tseng, Derek; Ozcan, Aydogan

    2015-03-01

    Giardia lamblia is a waterborne parasite that causes an intestinal infection, known as giardiasis, and it is found not only in countries with inadequate sanitation and unsafe water but also streams and lakes of developed countries. Simple, sensitive, and rapid detection of this pathogen is important for monitoring of drinking water. Here we present a cost-effective and field portable mobile-phone based fluorescence microscopy platform designed for automated detection of Giardia lamblia cysts in large volume water samples (i.e., 10 ml) to be used in low-resource field settings. This fluorescence microscope is integrated with a disposable water-sampling cassette, which is based on a flow-through porous polycarbonate membrane and provides a wide surface area for fluorescence imaging and enumeration of the captured Giardia cysts on the membrane. Water sample of interest, containing fluorescently labeled Giardia cysts, is introduced into the absorbent pads that are in contact with the membrane in the cassette by capillary action, which eliminates the need for electrically driven flow for sample processing. Our fluorescence microscope weighs ~170 grams in total and has all the components of a regular microscope, capable of detecting individual fluorescently labeled cysts under light-emitting-diode (LED) based excitation. Including all the sample preparation, labeling and imaging steps, the entire measurement takes less than one hour for a sample volume of 10 ml. This mobile phone based compact and cost-effective fluorescent imaging platform together with its machine learning based cyst counting interface is easy to use and can even work in resource limited and field settings for spatio-temporal monitoring of water quality.

  18. Advanced molecular devices based on light-driven molecular motors

    NARCIS (Netherlands)

    Chen, Jiawen

    2015-01-01

    Nature has provided a large collection of molecular machines and devices that are among the most amazing nanostructures on this planet. These machines are able to operate complex biological processes which are of great importance in our organisms. Inspired by these natural devices, artificial

  19. Multicolor Fluorescence Writing Based on Host-Guest Interactions and Force-Induced Fluorescence-Color Memory.

    Science.gov (United States)

    Matsunaga, Yuki; Yang, Jye-Shane

    2015-06-26

    A new strategy is reported for multicolor fluorescence writing on thin solid films with mechanical forces. This concept is illustrated by the use of a green-fluorescent pentiptycene derivative 1, which forms variably colored fluorescent exciplexes: a change from yellow to red was observed with anilines, and fluorescence quenching (a change to black) occurred in the presence of benzoquinone. Mechanical forces, such as grinding and shearing, induced a crystalline-to-amorphous phase transition in both the pristine and guest-adsorbed solids that led to a change in the fluorescence color (mechanofluorochromism) and a memory of the resulting color. Fluorescence drawings of five or more colors were created on glass or paper and could be readily erased by exposure to air and dichloromethane fumes. The structural and mechanistic aspects of the observations are also discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Synthesis and characterization of novel molecularly imprinted polymer - coated Mn-doped ZnS quantum dots for specific fluorescent recognition of cocaine.

    Science.gov (United States)

    Chantada-Vázquez, María Pilar; Sánchez-González, Juan; Peña-Vázquez, Elena; Tabernero, María Jesús; Bermejo, Ana María; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

    2016-01-15

    Mn-doped ZnS quantum dots (QDs) coated with a molecularly imprinted polymer (MIP) material selective toward cocaine and its metabolites have been prepared and applied to cocaine (COC) and metabolites assessment by spectrofluorimetry. Ultrasound irradiation (37kHz) was novelty used for performing the Mn-doped ZnS QDs synthesis as well as for preparing the QD based MIP-coated composite by precipitation polymerization (imprinting process). This fact allowed the synthesis to be accomplished in four hours. In addition, the use of ultrasound irradiation during MIP-QDs synthesis increased the homogeneity of the QDs size, and reduced nanoparticles agglomeration. MIP was synthesized using COC as a template molecule, ethylene dimethacrylate (EDMA) as a functional monomer, divinylbenzene (DVB) as a cross-linker, and 2,2'-azobisisobutyronitrile (AIBN) as an initiator. The fluorescence of MIP-coated QDs was quenched by the template (COC) and also by metabolites from COC such as benzoylecgonine (BZE), and ecgonine methyl ester (EME). Quenching was not observed when performing experiments with non-imprinted polymer (NIP)-coated QDs; and also, fluorescence quenching of MIP-coated QDs was not observed by other drugs of abuse and metabolites (heroin and cannabis abuse). This fact indicates that the prepared material recognize only COC (template) and metabolites. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Molecular basis for pH sensitivity and proton transfer in green fluorescent protein: protonation and conformational substates from electrostatic calculations.

    Science.gov (United States)

    Scharnagl, C; Raupp-Kossmann, R; Fischer, S F

    1999-10-01

    We performed a theoretical study to elucidate the coupling between protonation states and orientation of protein dipoles and buried water molecules in green fluorescent protein, a versatile biosensor for protein targeting. It is shown that the ionization equilibria of the wild-type green fluorescent protein-fluorophore and the internal proton-binding site E222 are mutually interdependent. Two acid-base transitions of the fluorophore occur in the presence of neutral (physiologic pH) and ionized (pH > 12) E222, respectively. In the pH-range from approximately 8 to approximately 11 ionized and neutral sites are present in constant ratio, linked by internal proton transfer. The results indicate that modulation of the internal proton sharing by structural fluctuations or chemical variations of aligning residues T203 and S65 cause drastic changes of the neutral/anionic ratio-despite similar physiologic fluorophore pK(a) s. Moreover, we find that dipolar heterogeneities in the internal hydrogen-bond network lead to distributed driving forces for excited-state proton transfer. A molecular model for the unrelaxed surrounding after deprotonation is discussed in relation to pathways providing fast ground-state recovery or slow stabilization of the anion. The calculated total free energy for excited-state deprotonation ( approximately 19 k(B)T) and ground-state reprotonation ( approximately 2 k(B)T) is in accordance with absorption and emission data (

  2. Instrumentation of Molecular Imaging on Site-Specific Targeting Fluorescent Peptide for Early Detection of Breast Cancer

    Science.gov (United States)

    Yu, Ping; Ma, Lixin

    2012-02-01

    In this work we developed two biomedical imaging techniques for early detection of breast cancer. Both image modalities provide molecular imaging capability to probe site-specific targeting dyes. The first technique, heterodyne CCD fluorescence mediated tomography, is a non-invasive biomedical imaging that uses fluorescent photons from the targeted dye on the tumor cells inside human breast tissue. The technique detects a large volume of tissue (20 cm) with a moderate resolution (1 mm) and provides the high sensitivity. The second technique, dual-band spectral-domain optical coherence tomography, is a high-resolution tissue imaging modality. It uses a low coherence interferometer to detect coherent photons hidden in the incoherent background. Due to the coherence detection, a high resolution (20 microns) is possible. We have finished prototype imaging systems for the development of both image modalities and performed imaging experiments on tumor tissues. The spectroscopic/tomographic images show contrasts of dense tumor tissues and tumor necrotic regions. In order to correlate the findings from our results, a diffusion-weighted magnetic resonance imaging (MRI) of the tumors was performed using a small animal 7-Telsa MRI and demonstrated excellent agreement.

  3. Efficient and Scalable Synthesis of 4-Carboxy-Pennsylvania Green Methyl Ester: A Hydrophobic Building Block for Fluorescent Molecular Probes.

    Science.gov (United States)

    Woydziak, Zachary R; Fu, Liqiang; Peterson, Blake R

    2014-01-01

    Fluorinated fluorophores are valuable tools for studies of biological systems. However, amine-reactive single-isomer derivatives of these compounds are often very expensive. To provide an inexpensive alternative, we report a practical synthesis of 4-carboxy-Pennsylvania Green methyl ester. Derivatives of this hydrophobic fluorinated fluorophore, a hybrid of the dyes Oregon Green and Tokyo Green, are often cell permeable, enabling labeling of intracellular targets and components. Moreover, the low pKa of Pennsylvania Green (4.8) confers bright fluorescence in acidic cellular compartments such as endosomes, enhancing its utility for chemical biology investigations. To improve access to the key intermediate 2,7-difluoro-3,6-dihydroxyxanthen-9-one, we subjected bis-(2,4,5-trifluorophenyl)methanone to iterative nucleophilic aromatic substitution by hydroxide on scales of > 40 g. This intermediate was used to prepare over 15 grams of pure 4-carboxy-Pennsylvania Green methyl ester in 28% overall yield without requiring chromatography. This compound can be converted into the amine reactive N -hydroxysuccinimidyl ester in essentially quantitative yield for the synthesis of a wide variety of fluorescent molecular probes.

  4. Molecular dynamics simulation studies of dielectric response and vibrational energy relaxation in photoactive yellow protein and green fluorescent protein

    Science.gov (United States)

    Xu, Yao; Gnanasekaran, Ramachandran; Leitner, David

    2012-02-01

    The first step in the photocycle of many proteins involves conformational change of a chromophore or a charge transfer reaction following photoexcitation. To explore the response of the protein and solvent environment to photoexcitation of the chromophore in photoactive yellow protein (PYP) and green fluorescent protein (GFP) we carried out molecular dynamics simulations of the dielectric response and vibrational energy relaxation (VER) from the chromophore to the protein and solvent. In PYP the time scale of the protein response, mainly contributed by Tyr42 and Glu46, to photoexcitation appears prominently between 0.1 and 0.3 picoseconds. The frequency-dependent VER rate also reveals dynamic coupling between the chromophore and residues that hydrogen bond to it. Resonances in the VER rate appear at frequencies comparable to the oscillations observed in recent fluorescence decay studies. In GFP, which undergoes excited state proton transfer about 10 ps following photoexcitation that may be assisted by specific chromophore vibrations, both the protein and water molecules inside the β-barrel surrounding the chromophore mediate the dielectric response.

  5. Interactions of perfluorooctanoic acid and perfluorooctanesulfonic acid with serum albumins by native mass spectrometry, fluorescence and molecular docking.

    Science.gov (United States)

    Chi, Quan; Li, Zhixiong; Huang, Juan; Ma, Jieyao; Wang, Xian

    2018-05-01

    The binding information of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) with bovine and human serum albumins was investigated and characterized in details by using a combination method of electrospray ionization mass spectrometry (ESI-MS), fluorescence, circular dichroism (CD) and molecular docking (MD). The ESI-MS analysis revealed that maximally eight PFOA or PFOS molecules could bind to serum albumins at high mole ratios of PFOA/PFOS. Association constants were measured by ESI-MS and suggested that PFOS had a better binding affinity than PFOA. PFOA and PFOS were likely to bind with serum albumins in more than one pocket. The CD data demonstrated that binding of PFOA and PFOS could change the conformation of serum albumins with decreasing α-helix content, which may affect the protein physiological function. The phenomenon of protein fluorescence quenching by the binding of PFOA and PFOS indicated that the hydrophobic pocket proximate to Trp 214 in human serum albumin might be one of the dominated binding sites. This assumption was further confirmed by MD simulation. Consistent to ESI-MS observation, MD results also displayed a stronger binding affinity of PFOS than PFOA according to the calculated binding free energy, which is probably ascribed to one more hydrogen bond formed in the PFOS-bound protein complexes. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. [The X-Ray Fluorescence Spectrometer Based on Pyroelectric Effect].

    Science.gov (United States)

    Dong, Yi-fan; Fan, Rui-rui; Guo, Dong-ya; Zhang, Chun-lei; Gao, Min; Wang, Jin-zhou; Liu, Ya-qing; Zhou, Da-wei; Wang, Huan-yu

    2016-02-01

    Pyroelectric X-ray generator is implemented, and an X-ray fluorescence spectrometer is accomplished by combining the pyroelectric X-ray generator with a high energy resolution silicon drift detector. Firstly, the parameters of the X-ray generator are decided by analyzing and calculating the influence of the thickness of the pyroelectriccrystal and the thickness of the target on emitted X-ray. Secondly, the emitted X-ray is measured. The energy of emitted X-ray is from 1 to 27 keV, containing the characteristic X-ray of Cu and Ta, and the max counting rate is more than 3 000 per second. The measurement also proves that the detector of the spectrometer has a high energy resolution which the FWMH is 210 eV at 8. 05 keV. Lastly, samples of Fe, Ti, Cr and high-Ti basalt are analyzed using the spectrometer, and the results are agreed with the elements of the samples. It shows that the spectrometer consisting of a pyroelectric X-ray generator and a silicon drift detector is effective for element analysis. Additionally, because each part of the spectrometer has a small volume, it can be easily modified to a portable one which is suitable for non-destructive, on-site and quick element analysis.

  7. Development of a novel deltamethrin sensor based on molecularly imprinted silica nanospheres embedded CdTe quantum dots

    Science.gov (United States)

    Ge, Shenguang; Lu, Juanjuan; Ge, Lei; Yan, Mei; Yu, Jinghua

    2011-09-01

    A novel procedure for the determination of deltmethrin (DM) is reported. The water-soluble CdTe quantum dots (QDs) and highly fluorescent silica molecularly imprinted nanospheres embedded CdTe QDs (CdTe-SiO 2-MIPs) were prepared and characterized by fluorescence spectroscopy, UV-vis spectroscopy, TEM and IR. The fluorescence nanosensor based CdTe-SiO 2-MIPs is developed. The possible quenching mechanism is discussed by DM. Under optimal conditions, the relative fluorescence intensity of CdTe-SiO 2-MIPs decreased with increasing DM by a Stern-Volmer type equation in the concentration range of 0.5-35.0 μg mL -1, the corresponding detection limit is 0.16 μg mL -1. The developed sensor based on CdTe-SiO 2-MIPs was applied to determine DM in fruit and vegetable samples.

  8. Basic amphipathic model peptides: Structural investigations in solution, studied by circular dichroism, fluorescence, analytical ultracentrifugation and molecular modelling

    Science.gov (United States)

    Mangavel, C.; Sy, D.; Reynaud, J. A.

    1999-05-01

    A twenty amino acid residue long amphipathic peptide made of ten leucine and ten lysine residues and four derivatives, in which a tryptophan, as a fluorescent probe, is substituted for a leucine, are studied. The peptides in water are mainly in an unordered conformation (~90%), and undergo a two state reversible transition upon heating, leading to a partially helical conformation (cold denaturation). Time resolved fluorescence results show that fluorescence decay for the four Trp containing peptides is best described by triple fluorescence decay kinetics. In TFE/water mixture, peptides adopt a single α-helix conformation but the Leu-Trp9 substitution leads to an effective helix destabilizing effect. In salted media, the peptides are fully helical and present a great tendency to self associate by bringing the hydrophobic faces of helices into close contact. This proceeds in non-cooperative multisteps leading to the formation of α helix aggregates with various degrees of complexation. Using modelling, the relative hydrophobic surface areas accessible to water molecules in n-mer structures are calculated and discussed. Nous avons étudié un peptide amphipathique composé de dix lysine et dix leucine, ainsi que quatre dérivés comportant un résidu tryptophane pour les études par fluorescence. Dans l'eau, les peptides ne sont pas structurés (~90%), et se structurent partiellement en hélice α par chauffage (dénaturation froide). Les mesures de déclin de fluorescence font apparaître une cinétique à trois temps de vie. Dans un mélange eau/TFE, les peptides adoptent une conformation en hélice α, mais la substitution Leu-Trp9 possède un effet déstabilisant. En mileu salin, les peptides sont totalement hélicoïdaux et ont tendance à s'agréger de façon à regrouper leur face hydrophobe. Ce processus se fait en plusieurs étapes avec des agrégats de taille variable. L'existence de tels agrégats est discutée sur la base de la modélisation mol

  9. Utilizing hyaluronic acid as a versatile platform for fluorescence resonance energy transfer-based glucose sensing.

    Science.gov (United States)

    Ge, Minghao; Bai, Pengli; Chen, Mingli; Tian, Jingjing; Hu, Jun; Zhi, Xu; Yin, Huancai; Yin, Jian

    2018-03-01

    Here, we utilized the ultrasonic emulsification technique to generate hyaluronic acid microspheres incorporating a fluorescence-based glucose biosensor. We synthesized a novel lanthanide ion luminophore based on Eu 3+ . Eu sulfosuccinimidyl dextran (Eu-dextran) and Alexa Fluor 647 sulfosuccinimidyl-ConA (Alexa Fluor 647-ConA) were encapsulated in hyaluronic acid hydrogel to generate microspheres. Glucose sensing was carried out using a fluorescence resonance energy transfer (FRET)-based assay principle. A proportional fluorescence intensity increase was found within a 0.5-10-mM glucose concentration range. The glucose-sensing strategy showed an excellent tolerance for potential interferents. Meanwhile, the fluorescent signal of hyaluronic acid microspheres was very stable after testing for 72 h in glucose solution. Overall, hyaluronic acid microspheres encapsulating sensing biomolecules offer a stable and biocompatible biosensor for a variety of applications including cell culture systems, tissue engineering, detection of blood glucose, etc. Graphical abstract We report an ingenious biosensor encapsulated in hyaluronic acid microspheres for monitoring of glucose. Glucose sensing is carried out using a fluorescence resonance energy transfer-based assay principle with a novel lanthanide ions luminophore. The glucose detection system has excellent biocompatibility and stability for monitoring of glucose.

  10. Hypoxia-Targeting Fluorescent Nanobodies for Optical Molecular Imaging of Pre-Invasive Breast Cancer

    NARCIS (Netherlands)

    van Brussel, Aram S A; Adams, Arthur; Oliveira, Sabrina; Dorresteijn, Bram; El Khattabi, Mohamed; Vermeulen, J. F.; van der Wall, Elsken; Mali, Willem P Th M; Derksen, Patrick W B; van Diest, Paul J; van Bergen En Henegouwen, Paul M P

    PURPOSE: The aim of this work was to develop a CAIX-specific nanobody conjugated to IRDye800CW for molecular imaging of pre-invasive breast cancer. PROCEDURES: CAIX-specific nanobodies were selected using a modified phage display technology, conjugated site-specifically to IRDye800CW and evaluated

  11. Hypoxia-Targeting Fluorescent Nanobodies for Optical Molecular Imaging of Pre-Invasive Breast Cancer

    NARCIS (Netherlands)

    van Brussel, Aram S A; Adams, Arthur; Oliveira, Sabrina; Dorresteijn, Bram; El Khattabi, Mohamed; Vermeulen, Jeroen F.; van der Wall, Elsken; Mali, W.P.T.M.; Derksen, Patrick W B; van Diest, Paul J.; van Bergen En Henegouwen, Paul M P

    Purpose: The aim of this work was to develop a CAIX-specific nanobody conjugated to IRDye800CW for molecular imaging of pre-invasive breast cancer. Procedures: CAIX-specific nanobodies were selected using a modified phage display technology, conjugated site-specifically to IRDye800CW and evaluated

  12. A Cu(II)-benzoyl hydrazone based fluorescent probe for lipopolysaccharides

    International Nuclear Information System (INIS)

    Yang, Jing; Huang, Lei; Guo, Zhengyu; Ren, Wang; Wang, Qiusheng

    2016-01-01

    A novel fluorescent compound 7-diethylamino-3-[4'-N-((2-ethoxy)ethanol)methyl aminobenzoylhydrazone]methyl coumarin (I) was synthesized and employed as a fluorescent probe for detecting lipopolysaccharides (LPS). This coumarin-based probe exhibited high fluorescence in aqueous environment, which could be quenched in the presence of Cu 2+ ion due to the complexation between I and Cu 2+ . However, the fluorescence of I can be recovered upon addition of LPS, because the binding between LPS and Cu 2+ ion is stronger than that between I and Cu 2+ ion, which disassociates the complexation of I and Cu 2+ ion and releases the fluorescence of I. Therefore, this three-component sensing system can be used to detect lipopolysaccharides in a facile manner. In addition, biological imaging studies have demonstrated that the probe can be used to evaluate the biosorptive capacity for Cu 2+ ion in living bacteria. - Highlights: • A novel water-soluble coumarin derivative was synthesized. • Compound I was used to detect lipopolysaccharides selectively. • The detection process was studied upon UV–vis and fluorescence spectrum. • The bioimaging application of I in bacteria was studied.

  13. Use of Time-Resolved Fluorescence to Monitor Bioactive Compounds in Plant Based Foodstuffs

    Directory of Open Access Journals (Sweden)

    M. Adília Lemos

    2015-06-01

    Full Text Available The study of compounds that exhibit antioxidant activity has recently received much interest in the food industry because of their potential health benefits. Most of these compounds are plant based, such as polyphenolics and carotenoids, and there is a need to monitor them from the field through processing and into the body. Ideally, a monitoring technique should be non-invasive with the potential for remote capabilities. The application of the phenomenon of fluorescence has proved to be well suited, as many plant associated compounds exhibit fluorescence. The photophysical behaviour of fluorescent molecules is also highly dependent on their microenvironment, making them suitable probes to monitor changes in pH, viscosity and polarity, for example. Time-resolved fluorescence techniques have recently come to the fore, as they offer the ability to obtain more information, coupled with the fact that the fluorescence lifetime is an absolute measure, while steady state just provides relative and average information. In this work, we will present illustrative time-resolved measurements, rather than a comprehensive review, to show the potential of time-resolved fluorescence applied to the study of bioactive substances. The aim is to help assess if any changes occur in their form, going from extraction via storage and cooking to the interaction with serum albumin, a principal blood transport protein.

  14. [Lake algae chemotaxonomy technology based on fluorescence excitation emission matrix and parallel factor analysis].

    Science.gov (United States)

    Chen, Xiao-Na; Han, Xiu-Rong; Su, Rong-Guo; Shi, Xiao-Yong

    2014-03-01

    An in vivo three-dimensional fluorescence method for the determination of algae community structure was developed by parallel factor (PARAFAC) analysis and CHEMTAX. The PARAFAC model was applied to fluorescence excitation-emission matrix (EEM) of 23 algae species and 12 fluorescent components were identified according to the residual sum of squares and specificity of the composition profiles of fluorescent. Based on the 12 fluorescent components, the algae species at different growth stages were correctly classified at the division level using Bayesian discriminant analysis (BDA). Then the reference fluorescent component ratio matrix was constructed for CHEMTAX, and the EEM-PARAFAC-CHEMTAX method was developed to differentiate taxonomic groups of algae. When the fluorometric method was used for 531 single-species samples, the average correct discrimination ratio (CDR) was 99.1% and the correct discrimination ratios (CDRs) were 100% at the division level except Chlorophyta, the CDR of which was 97.5%. The CDRs for 95 mixtures were above 98.5% for the dominant algae species and above 90.5% for the subdominant algae species, with average relative contents of 69.7% and 26.4%, respectively. This technique would be of great aid when low-cost and rapid analysis is needed for samples in a large batch.

  15. Gold-based hybrid nanomaterials for biosensing and molecular diagnostic applications.

    Science.gov (United States)

    Kim, Jung Eun; Choi, Ji Hye; Colas, Marion; Kim, Dong Ha; Lee, Hyukjin

    2016-06-15

    The properties of gold nanomaterials are particularly of interest to many researchers, since they show unique physiochemical properties such as optical adsorption of specific wavelength of light, high electrical conductance with rich surface electrons, and facile surface modification with sulfhydryl groups. These properties have facilitated the use of gold nanomaterials in the development of various hybrid systems for biosensors and molecular diagnostics. Combined with various synthetic materials such as fluorescence dyes, polymers, oligonucleotides, graphene oxides (GO), and quantum dots (QDs), the gold-based hybrid nanomaterials offer multi-functionalities in molecular detection with high specificity and sensitivity. These two aspects result in the increase of detection speed as well as the lower detection limits, having shown that this diagnosis method is more effective than other conventional ones. In this review, we have highlighted various examples of nanomaterials for biosensing and molecular diagnostics. The gold-based hybrid systems are categorized by three distinct detection approaches, in which include (1) optical, such as surface plasmon resonance (SPR), RAMAN, and surface-enhanced Raman scattering (SERS), (2) fluorescence, such as förster resonance energy transfer (FRET) and nanomaterial surface energy transfer (NSET), and (3) electrochemical, such as potentiometic, amperometric, and conductometric. Each example provides the detailed mechanism of molecular detection as well as the supporting experimental result with the limit of detection (LOD). Lastly, future perspective on novel development of gold-based hybrid nanomaterials is discussed as well as their challenges. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Novel chitosan-based fluorescent materials for the selective detection and adsorption of Fe3+ in water and consequent bio-imaging applications.

    Science.gov (United States)

    Meng, Qingtao; Su, Weiping; He, Cheng; Duan, Chunying

    2012-08-15

    A series of fluorescent materials were synthesized by modification of chitosan (CS) with 4-fluoresceincarboxaldehyde (Fluo) and N-methyl-carbazole-3-aldehyde (Cb). Both L-CS-Fluo and L-CS-Cb feature excellent water-solubility and exhibit highly selective fluorescence response to Fe(3+) in environment and biological fields. The high-molecular weight chitosan-based materials: H-CS-Fluo and H-CS-Cb take on doubled absorptivity compared to free chitosan due to the introduction of fluorescence probes. These modified-probe chitosan materials could be regenerated by treating with EDTA. Meanwhile, monomer probes AG-Fluo and AG-Cb linked by D-glucosamine were also synthesized to explore the binding efficiency. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Using Fluorescent Viruses for Detecting Bacteria in Water

    Science.gov (United States)

    Tabacco, Mary Beth; Qian, Xiaohua; Russo, Jaimie A.

    2009-01-01

    A method of detecting water-borne pathogenic bacteria is based partly on established molecular-recognition and fluorescent-labeling concepts, according to which bacteria of a species of interest are labeled with fluorescent reporter molecules and the bacteria can then be detected by fluorescence spectroscopy. The novelty of the present method lies in the use of bacteriophages (viruses that infect bacteria) to deliver the fluorescent reporter molecules to the bacteria of the species of interest.

  18. A coumarin based Schiff base probe for selective fluorescence detection of Al3 + and its application in live cell imaging

    Science.gov (United States)

    Sen, Bhaskar; Sheet, Sanjoy Kumar; Thounaojam, Romita; Jamatia, Ramen; Pal, Amarta Kumar; Aguan, Kripamoy; Khatua, Snehadrinarayan

    2017-02-01

    A new coumarin based Schiff base compound, CSB-1 has been synthesized to detect metal ion based on the chelation enhanced fluorescence (CHEF). The cation binding properties of CSB-1 was thoroughly examined in UV-vis and fluorescence spectroscopy. In fluorescence spectroscopy the compound showed high selectivity toward Al3 + ion and the Al3 + can be quantified in mixed aqueous buffer solution (MeOH: 0.01 M HEPES Buffer; 9:1; v/v) at pH 7.4 as well as in BSA media. The fluorescence intensity of CSB-1 was enhanced by 24 fold after addition of only five equivalents of Al3 +. The fluorescence titration of CSB-1 with Al3 + in mixed aqueous buffer afforded a binding constant, Ka = (1.06 ± 0.2) × 104 M- 1. The colour change from light yellow to colourless and the appearance of blue fluorescence, which can be observed by the naked eye, provides a real-time method for Al3 + sensing. Further the live cell imaging study indicated that the detection of intracellular Al3 + ions are also readily possible in living cell.

  19. A new rhodamine-based fluorescent chemodosimeter for mercuric ions in water media.

    Science.gov (United States)

    Quy, Phan Tu; Hien, Nguyen Khoa; Bao, Nguyen Chi; Nhan, Doan Thanh; Khanh, Dang Van; Nhung, Nguyen Thi Ai; Tung, Truong Quy; Luyen, Nguyen Dinh; Quang, Duong Tuan

    2015-05-01

    A new rhodamine-ethylenediamine-nitrothiourea conjugate (RT) was synthesized and its sensing property as a fluorescent chemodosimeter toward metal ions was explored in water media. Analytical results from absorption and fluorescence spectra revealed that the addition of Hg(2+) ions to the aqueous solution of the chemodosimeter RT caused a distinct fluorescence OFF-ON response with a remarkable visual color change from colorless to pink; however, no clear spectral and color changes were observed from other metal ions including: Zn(2+) , Cu(2+) , Cd(2+) , Pb(2+) , Ag(+) , Fe(2+) , Cr(3+) , Co(3+) , Ni(2+) , Ca(2+) , Mg(2+) , K(+) and Na(+) . The sensing results and the molecular structure suggested that a Hg(2+) -induced a desulfurization reaction and cyclic guanylation of the thiourea moiety followed by ring-opening of the rhodamine spirolactam in RT are responsible for a distinct fluorescence turn-on signal, indicating that RT is a remarkably sensitive and selective chemodosimeter for Hg(2+) ions in aqueous solution. Hg(2+) within a concentration range from 0.1 to 25 μM can be determined using RT as a chemodosimeter and a detection limit of 0.04 μM is achieved. Copyright © 2014 John Wiley & Sons, Ltd.

  20. A coumarin-based fluorescent probe as a central nervous system disease biomarker.

    Science.gov (United States)

    Yap, Ann-Chee; Mahamad, Ummi Affah; Lim, Shen-Yang; Kim, Hae-Jo; Choo, Yeun-Mun

    2014-11-10

    Homocysteine and methylmalonic acid are important biomarkers for diseases associated with an impaired central nervous system (CNS). A new chemoassay utilizing coumarin-based fluorescent probe 1 to detect the levels of homocysteine is successfully implemented using Parkinson's disease (PD) patients' blood serum. In addition, a rapid identification of homocysteine and methylmalonic acid levels in blood serum of PD patients was also performed using the liquid chromatography-mass spectrometry (LC-MS). The results obtained from both analyses were in agreement. The new chemoassay utilizing coumarin-based fluorescent probe 1 offers a cost- and time-effective method to identify the biomarkers in CNS patients.

  1. A Coumarin-Based Fluorescent Probe as a Central Nervous System Disease Biomarker

    Directory of Open Access Journals (Sweden)

    Ann-Chee Yap

    2014-11-01

    Full Text Available Homocysteine and methylmalonic acid are important biomarkers for diseases associated with an impaired central nervous system (CNS. A new chemoassay utilizing coumarin-based fluorescent probe 1 to detect the levels of homocysteine is successfully implemented using Parkinson’s disease (PD patients’ blood serum. In addition, a rapid identification of homocysteine and methylmalonic acid levels in blood serum of PD patients was also performed using the liquid chromatography-mass spectrometry (LC-MS. The results obtained from both analyses were in agreement. The new chemoassay utilizing coumarin-based fluorescent probe 1 offers a cost- and time-effective method to identify the biomarkers in CNS patients.

  2. Controllable molecular aggregation and fluorescence properties of 1,3,4-oxadiazole derivative

    KAUST Repository

    Li, Min

    2015-10-14

    The molecular self-assembly behaviour of 2,2’-Bis-(4-hexyloxyphenyl)-bi-1,3,4-oxadiazole (BOXD-6) in solution, on surfaces and in bulk crystals, and its photo-physical properties were studied via a combination of experimental techniques and theoretical calculations. It is found that BOXD-6 molecules self-assemble into both H- and J-aggregates at moderate concentration (~10-4 M) and then transit to exclusive J-aggregates at higher concentration (~10-3 M) in tetrahydrofuran. In H-aggregation (α polymorph), BOXD-6 adopts a linear conformation and forms a one- dimensional layered structure; in J-aggregation (β polymorph), it adopts a Z-shaped conformation and form a more ordered two-dimensional layered structure. A π-stacking structure is observed in both cases, and adjacent molecules in the J-aggregation show larger displacement along the molecular long axis direction than that in H-aggregation. Although J-aggregates are almost the only component in concentrated solutions (10-3 M), both H- and J-aggregates can be obtained if concentrated solution is transformed onto substrates through a simple drop-casting method. Such a phase transition during film formation can be easily avoided by adding water as precipitator; a film with pure J-aggregates is then obtained. In order to get more information on molecular self-assembly, intermolecular interaction potential energy surfaces (PES) were evaluated via theoretical calculations at the DFT level (M062x/6-31G**). The PES not only confirm the molecular stacking structures found in crystals but also predict some other likely structures, which will be the target of future experiments.

  3. Evaluation of a fluorescence-based method for antibabesial drug screening.

    Science.gov (United States)

    Guswanto, Azirwan; Sivakumar, Thillaiampalam; Rizk, Mohamed Abdo; Elsayed, Shimaa Abd Elsalam; Youssef, Mohamed Ahmed; ElSaid, ElSaid El Shirbini; Yokoyama, Naoaki; Igarashi, Ikuo

    2014-08-01

    In vitro evaluation of chemotherapeutic agents against Babesia and Theileria parasites has become routine, and the effectiveness of these chemicals is usually determined by comparing the parasitemia dynamics of untreated and treated parasites. Although microscopy is widely used to calculate parasitemia, several disadvantages are associated with this technique. The present study evaluated a fluorescence-based method using SYBR green I stain (SG I) to screen antibabesial agents in in vitro cultures of Babesia bovis. The linearity between relative fluorescence units (RFU) and parasitemia was found to be well correlated with a 0.9944 goodness-of-fit (r(2)) value. Subsequently, 50% inhibitory concentration (IC50) values were calculated for 3 antiprotozoan agents, diminazene aceturate, nimbolide, and gedunin, by this method. For diminazene aceturate and nimbolide, the IC(50)s determined by the fluorescence-based method (408 nM and 8.13 μM, respectively) and microscopy (400.3 nM and 9.4 μM, respectively) were in agreement. Furthermore, the IC50 of gedunin determined by the fluorescence-based method (19 μM) was similar to the recently described microscopy-based value (21.7 μM) for B. bovis. Additionally, the Z' factor (0.80 to 0.90), signal-to-noise (S/N) ratio (44.15 to 87.64), coefficient of variation at the maximum signal (%CVmax) (0.50 to 2.85), and coefficient of variation at the minimum signal (%CVmin) (1.23 to 2.21) calculated for the fluorescence method using diminazene aceturate were comparable to those previously determined in malaria research for this assay. These findings suggest that the fluorescence-based method might be useful for antibabesial drug screening and may have potential to be developed into a high-throughput screening (HTS) assay. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  4. NIR-Cyanine Dye Linker: a Promising Candidate for Isochronic Fluorescence Imaging in Molecular Cancer Diagnostics and Therapy Monitoring.

    Science.gov (United States)

    Komljenovic, Dorde; Wiessler, Manfred; Waldeck, Waldemar; Ehemann, Volker; Pipkorn, Ruediger; Schrenk, Hans-Hermann; Debus, Jürgen; Braun, Klaus

    2016-01-01

    Personalized anti-cancer medicine is boosted by the recent development of molecular diagnostics and molecularly targeted drugs requiring rapid and efficient ligation routes. Here, we present a novel approach to synthetize a conjugate able to act simultaneously as an imaging and as a chemotherapeutic agent by coupling functional peptides employing solid phase peptide synthesis technologies. Development and the first synthesis of a fluorescent dye with similarity in the polymethine part of the Cy7 molecule whose indolenine-N residues were substituted with a propylene linker are described. Methylating agent temozolomide is functionalized with a tetrazine as a diene component whereas Cy7-cell penetrating peptide conjugate acts as a dienophilic reaction partner for the inverse Diels-Alder click chemistry-mediated ligation route yielding a theranostic conjugate, 3-mercapto-propionic-cyclohexenyl-Cy7-bis-temozolomide-bromide-cell penetrating peptide. Synthesis route described here may facilitate targeted delivery of the therapeutic compound to achieve sufficient local concentrations at the target site or tissue. Its versatility allows a choice of adequate imaging tags applicable in e.g. PET, SPECT, CT, near-infrared imaging, and therapeutic substances including cytotoxic agents. Imaging tags and therapeutics may be simultaneously bound to the conjugate applying click chemistry. Theranostic compound presented here offers a solid basis for a further improvement of cancer management in a precise, patient-specific manner.

  5. A fluorescence turn-on chemosensor for hydrogen sulfate anion based on quinoline and naphthalimide

    Science.gov (United States)

    Luo, Zaoli; Yin, Kai; Yu, Zhu; Chen, Mengxue; Li, Yan; Ren, Jun

    2016-12-01

    A new fluorescence turn-on chemosensor 1 based on quinoline and naphthalimide was prepared and its anion sensing toward various anions behavior was explored in this paper. Sensor 1 exhibited a highly selective fluorescent response toward HSO4- with an 8-fold fluorescence intensity enhancement in the presence of 10 equiv. of HSO4- in DMSO-H2O (1/1, v/v) solution. The sensor also displayed high sensitivity to hydrogen sulfate and the detection limit was calculated to be 7.79 × 10- 7 M. The sensing mechanism has been suggested to proceed via a hydrolysis process of the Schiff base group. The hydrolysis product has been isolated and further identified by 1H NMR and MS.

  6. Molecularly imprinted polymers prepared using protein-conjugated cleavable monomers followed by site-specific post-imprinting introduction of fluorescent reporter molecules.

    Science.gov (United States)

    Suga, Yusuke; Sunayama, Hirobumi; Ooya, Tooru; Takeuchi, Toshifumi

    2013-10-04

    Molecularly imprinted polymers were prepared using a protein-conjugated disulfide cleavable monomer. After removing the protein by disulfide reduction, a thiol-reactive fluorophore was introduced into the thiol residue located only inside the imprinted cavity, resulting in specific transduction of the binding events into fluorescence spectral change.

  7. Illuminating the origins of spectral properties of green fluorescent proteins via proteochemometric and molecular modeling.

    Science.gov (United States)

    Nantasenamat, Chanin; Simeon, Saw; Owasirikul, Wiwat; Songtawee, Napat; Lapins, Maris; Prachayasittikul, Virapong; Wikberg, Jarl E S

    2014-10-15

    Green fluorescent protein (GFP) has immense utility in biomedical imaging owing to its autofluorescent nature. In efforts to broaden the spectral diversity of GFP, there have been several reports of engineered mutants via rational design and random mutagenesis. Understanding the origins of spectral properties of GFP could be achieved by means of investigating its structure-activity relationship. The first quantitative structure-property relationship study for modeling the spectral properties, particularly the excitation and emission maximas, of GFP was previously proposed by us some years ago in which quantum chemical descriptors were used for model development. However, such simplified model does not consider possible effects that neighboring amino acids have on the conjugated π-system of GFP chromophore. This study describes the development of a unified proteochemometric model in which the GFP chromophore and amino acids in its vicinity are both considered in the same model. The predictive performance of the model was verified by internal and external validation as well as Y-scrambling. Our strategy provides a general solution for elucidating the contribution that specific ligand and protein descriptors have on the investigated spectral property, which may be useful in engineering novel GFP variants with desired characteristics. Copyright © 2014 Wiley Periodicals, Inc.

  8. Fluorescence enhancement of the conjugated polymer films based on well-ordered Au nanoparticle arrays

    Energy Technology Data Exchange (ETDEWEB)

    Zhong, Benbin; Zu, Xihong, E-mail: zxhong329@126.com; Yi, Guobin, E-mail: ygb702@163.com; Huang, Hailiang; Zhang, Minghai; Luo, Hongsheng [Guangdong University of Technology, School of Chemical Engineering and Light Industry (China)

    2016-09-15

    In this paper, well-ordered Au nanoparticle arrays on silicon substrates were employed as efficient metal-enhanced fluorescence (MEF) substrates for investigating the fluorescence properties of the conjugated polymer poly(3-hexylthiophene) (P3HT). The ordered Au nanoparticle arrays were fabricated by block copolymer self-assembly technology, and the particle sizes were controlled by adjusting the molar ratios of HAuCl{sub 4} precursor to vinyl pyridine units. The approach is economical and suitable to fabricate large-area MEF substrates. The results about fluorescence properties of P3HT showed that the fluorescence intensities of the P3HT films were improved on ordered Au nanoparticle arrays compared to those on bare silicon substrate and were significantly enhanced with the Au nanoparticle sizes increasing. The mechanism is based on localized surface plasmon resonances, coupling and propagating surface plasmons, and the emission enhancement mainly resulted from the increase of the excitation rate. This work provides a new way to prepare efficient MEF substrates for high-performance fluorescence-based devices.

  9. Fluorescence enhancement of the conjugated polymer films based on well-ordered Au nanoparticle arrays

    International Nuclear Information System (INIS)

    Zhong, Benbin; Zu, Xihong; Yi, Guobin; Huang, Hailiang; Zhang, Minghai; Luo, Hongsheng

    2016-01-01

    In this paper, well-ordered Au nanoparticle arrays on silicon substrates were employed as efficient metal-enhanced fluorescence (MEF) substrates for investigating the fluorescence properties of the conjugated polymer poly(3-hexylthiophene) (P3HT). The ordered Au nanoparticle arrays were fabricated by block copolymer self-assembly technology, and the particle sizes were controlled by adjusting the molar ratios of HAuCl 4 precursor to vinyl pyridine units. The approach is economical and suitable to fabricate large-area MEF substrates. The results about fluorescence properties of P3HT showed that the fluorescence intensities of the P3HT films were improved on ordered Au nanoparticle arrays compared to those on bare silicon substrate and were significantly enhanced with the Au nanoparticle sizes increasing. The mechanism is based on localized surface plasmon resonances, coupling and propagating surface plasmons, and the emission enhancement mainly resulted from the increase of the excitation rate. This work provides a new way to prepare efficient MEF substrates for high-performance fluorescence-based devices.

  10. Synthesis and application of a highly selective copper ions fluorescent probe based on the coumarin group

    Science.gov (United States)

    He, Guangjie; Liu, Xiangli; Xu, Jinhe; Ji, Liguo; Yang, Linlin; Fan, Aiying; Wang, Songjun; Wang, Qingzhi

    2018-02-01

    A highly selective copper ions fluorescent probe based on the coumarin-type Schiff base derivative 1 (probe) was produced by condensation reaction between coumarin carbohydrazide and 1H-indazole-3-carbaldehyde. The UV-vis spectroscopy showed that the maximum absorption peak of compound 1 appeared at 439 nm. In the presence of Cu2 + ions, the maximum peak decreased remarkably compared with other physiological important metal ions and a new absorption peak at 500 nm appeared. The job's plot experiments showed that complexes of 1:2 binding mode were formed in CH3CN:HEPES (3:2, v/v) solution. Compound 1 exhibited a strong blue fluorescence. Upon addition of copper ions, the fluorescence gradually decreased and reached a plateau with the fluorescence quenching rate up to 98.73%. The detection limit for Cu2 + ions was estimated to 0.384 ppm. Fluorescent microscopy experiments demonstrated that probe 1 had potential to be used to investigate biological processes involving Cu2 + ions within living cells.

  11. Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex

    Directory of Open Access Journals (Sweden)

    Pilar Santolaria

    2016-01-01

    Full Text Available This study was designed to determine the ability of computer-assisted sperm morphometry analysis (CASA-Morph with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa (SX and SY, respectively. Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average (P < 0.001 although with important differences between bulls. A simultaneous evaluation of all the measured features by discriminant analysis revealed that nuclear area and average fluorescence intensity were the variables selected by stepwise discriminant function analysis as the best discriminators between SX and SY. In the second experiment, the sperm nuclear morphometric results from CASA-Morph in nonsexed (mixed SX and SY and sexed (SX semen samples from four bulls were compared. FISH allowed a successful classification of spermatozoa according to their sex chromosome content. X-sexed spermatozoa displayed a larger size and fluorescence intensity than nonsexed spermatozoa (P < 0.05. We conclude that the CASA-Morph fluorescence-based method has the potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.

  12. Emergence of White Organic Light-Emitting Diodes Based on Thermally Activated Delayed Fluorescence

    Directory of Open Access Journals (Sweden)

    Peng Xiao

    2018-02-01

    Full Text Available Recently, thermally activated delayed fluorescence (TADF organic light-emitting diodes (OLEDs have attracted both academic and industrial interest due to their extraordinary characteristics, such as high efficiency, low driving voltage, bright luminance, lower power consumption and potentially long lifetime. In this invited review, the fundamental concepts of TADF have been firstly introduced. Then, main approaches to realize WOLEDs based on TADF have been summarized. More specifically, the recent development of WOLEDs based on all TADF emitters, WOLEDs based on TADF and conventional fluorescence emitters, hybrid WOLEDs based on blue TADF and phosphorescence emitters and WOLEDs based on TADF exciplex host and phosphorescence dopants is highlighted. In particular, design strategies, device structures, working mechanisms and electroluminescent processes of the representative WOLEDs based on TADF are reviewed. Finally, challenges and opportunities for further enhancement of the performance of WOLEDs based on TADF are presented.

  13. Nanoscopic description of biomembrane electrostatics: results of molecular dynamics simulations and fluorescence probing.

    Science.gov (United States)

    Demchenko, Alexander P; Yesylevskyy, Semen O

    2009-08-01

    Electrostatic fields generated on and inside biological membranes are recognized to play a fundamental role in key processes of cell functioning. Their understanding requires an adequate description on the level of elementary charges and the reconstruction of electrostatic potentials by integration over all elementary interactions. Out of all the available research tools, only molecular dynamics simulations are capable of this, extending from the atomic to the mesoscopic level of description on the required time and space scale. A complementary approach is that offered by molecular probe methods, with the application of electrochromic dyes. Highly sensitive to intermolecular interactions, they generate integrated signals arising from electric fields produced by elementary charges at the sites of their location. This review is an attempt to provide a critical analysis of these two approaches and their present and potential applications. The results obtained by both methods are consistent in that they both show an extremely complex profile of the electric field in the membrane. The nanoscopic view, with two-dimensional averaging over the bilayer plane and formal separation of the electrostatic potential into surface (Psi(s)), dipole (Psi(d)) and transmembrane (Psi(t)) potentials, is constructive in the analysis of different functional properties of membranes.

  14. Detection of substrate binding of a collagen-specific molecular chaperone HSP47 in solution using fluorescence correlation spectroscopy.

    Science.gov (United States)

    Kitamura, Akira; Ishida, Yoshihito; Kubota, Hiroshi; Pack, Chan-Gi; Homma, Takayuki; Ito, Shinya; Araki, Kazutaka; Kinjo, Masataka; Nagata, Kazuhiro

    2018-02-26

    Heat shock protein 47 kDa (HSP47), an ER-resident and collagen-specific molecular chaperone, recognizes collagenous hydrophobic amino acid sequences (Gly-Pro-Hyp) and assists in secretion of correctly folded collagen. Elevated collagen production is correlated with HSP47 expression in various diseases, including fibrosis and keloid. HSP47 knockdown ameliorates liver fibrosis by inhibiting collagen secretion, and inhibition of the interaction of HSP47 with procollagen also prevents collagen secretion. Therefore, a high-throughput system for screening of drugs capable of inhibiting the interaction between HSP47 and collagen would aid the development of novel therapies for fibrotic diseases. In this study, we established a straightforward method for rapidly and quantitatively measuring the interaction between HSP47 and collagen in solution using fluorescence correlation spectroscopy (FCS). The diffusion rate of HSP47 labeled with Alexa Fluor 488 (HSP47-AF), a green fluorescent dye, decreased upon addition of type I or III collagen, whereas that of dye-labeled protein disulfide isomerase (PDI) or bovine serum albumin (BSA) did not, indicating that specific binding of HSP47 to collagen could be detected using FCS. Using this method, we calculated the dissociation constant of the interaction between HSP47 and collagen. The binding ratio between HSP47-AF and collagen did not change in the presence of sodium chloride, confirming that the interaction was hydrophobic in nature. In addition, we observed dissociation of collagen from HSP47 at low pH and re-association after recovery to neutral pH. These observations indicate that this system is appropriate for detecting the interaction between HSP47 and collagen, and could be applied to high-throughput screening for drugs capable of suppressing and/or curing fibrosis. Copyright © 2018 Elsevier Inc. All rights reserved.

  15. Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay

    Science.gov (United States)

    Xing, Yun-Peng; Liu, Chun; Zhou, Xiao-Hong; Shi, Han-Chang

    2015-01-01

    This work was the first to report that the kanamycin-binding DNA aptamer (5'-TGG GGG TTG AGG CTA AGC CGA-3') can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNA-TO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59 nM with a linear working range of 0.1 μM to 20 μM for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future.

  16. A sol-gel based molecular imprint incorporating carbon dots for fluorometric determination of nicotinic acid

    International Nuclear Information System (INIS)

    Zuo, Pengli; Gao, Junfa; Liu, Jianha; Zhao, Mingming; Zhao, Jiahong; Peng, Jun; Zuo, Pengjian; He, Hua

    2016-01-01

    We are introducing functionalized carbon dots (C-dots) coated with a shell of molecularly imprinted sol-gel as a new tool in molecular imprint-based detection. Specifically, an imprint recognizing nicotinic acid (NA) was prepared in two steps. The first involves pyrolytic decomposition of citric acid in the presence of aminopropyltriethoxysilane to yield triethoxysilyl-modified C-dots with a typical size of 2.8 ± 1.1 nm. These are then polycondensed in the presence of tetraethoxysilane and NA at room temperature to give spherical silica nanoparticles (SiNPs) with a typical size of ∼300 nm and containing C-dots and NA in the silica matrix. NA was then removed by extraction. The resulting SiNPs are well permeable to NA, photostable, display strong blue luminescence and can bind NA fairly selectively. The fluorometric detection scheme is based on the finding that increasing concentrations of NA quench the fluorescence of the C-dots in the SiNPs. NA can be determined by this method in the 0.5 to 10.5 μM concentration range, with a 12.6 nM detection limit. The composite was successfully utilized as a fluorescent probe for the determination of NA in spiked human urine samples. The method is believed to have a wider scope in being applicable to other analytes that are capable of quenching the fluorescence of C-dots. (author)

  17. New approach to breast tumor detection based on fluorescence x-ray analysis

    Directory of Open Access Journals (Sweden)

    Okuyama, Fumio

    2010-01-01

    Full Text Available A new technical approach to breast-tumor detection is proposed. The technique is based on fluorescence x-ray analysis, and can identify a miniature malignant tumor within the breast. The primary beam intensity needed in fluorescence x-ray analysis is on a lower order of magnitude than that used in mammography. Thus, the newly-proposed technique would enable detection of a still tiny breast cancer while dramatically lowering the radiation dose. Field-emission x-ray sources might be a key for translating this concept into a medical technique.

  18. Compound parabolic concentrator optical fiber tip for FRET-based fluorescent sensors

    DEFF Research Database (Denmark)

    Hassan, Hafeez Ul; Nielsen, Kristian; Aasmul, Soren

    2015-01-01

    polymer optical fiber for an excitation and emission wavelength of 550 nm and 650nm, respectively. The model suggests an increase of a factor of 1.6 to 4 in the collected fluorescent power for an ideal CPC tip, as compared to the plane-cut fiber tip for fiber lengths between 5 and 45mm......The Compound Parabolic Concentrator (CPC) optical fiber tip shape has been proposed for intensity based fluorescent sensors working on the principle of FRET (Förster Resonance Energy Transfer). A simple numerical Zemax model has been used to optimize the CPC tip geometry for a step-index multimode...

  19. Laser-induced absorption and fluorescence studies of photochromic Schiff bases

    DEFF Research Database (Denmark)

    Kownacki, K.; Mordzinski, A.; Wilbrandt, R.

    1994-01-01

    Three photochromic Schiff bases: N-salicylideneaniline (SA), N-salicylidene-1-naphthylamine (SN), and N,N-bis-(salicylidene)-p-phenylenediamine (Bsp), were studied in acetonitrile by means of steady-state and time-resolved absorption and fluorescence spectroscopy, as well as semiempirical quantum...... chemical calculations. In all these molecules, the transient absorption and two-step laser-induced fluorescence spectra of long-lived transients are remarkably similar. The photochromic species is tentatively assigned to the non-hydrogen bonded form of the proton transfer reaction product...

  20. Microprocessor-based system for automatic X-ray diffraction and fluorescence

    International Nuclear Information System (INIS)

    Souza, A.M. de; Carmo, L.C.S. do; Pereira, V.J.E.; Soares, E.A.

    1984-01-01

    A data acquisition and processing device appropriate for X-ray analysis and goniometer control was built. The Z-80 based system as well as the whole architeture is described. The advantages and new possibilities of the automated instrument as compared to the traditional ones are listed. The X-ray diffraction and fluorescence techniques can take advantage of the automation. (Author) [pt

  1. A Fundamental Parameter-Based Calibration Model for an Intrinsic Germanium X-Ray Fluorescence Spectrometer

    DEFF Research Database (Denmark)

    Christensen, Leif Højslet; Pind, Niels

    1982-01-01

    A matrix-independent fundamental parameter-based calibration model for an energy-dispersive X-ray fluorescence spectrometer has been developed. This model, which is part of a fundamental parameter approach quantification method, accounts for both the excitation and detection probability. For each...

  2. A fluorescence resonance energy transfer-based method for histone methyltransferases

    DEFF Research Database (Denmark)

    Devkota, Kanchan; Lohse, Brian; Nyby Jakobsen, Camilla

    2015-01-01

    A simple dye–quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye...

  3. Gauging and visualizing c-di-GMP levels in pseudomonas aeruginosa using fluorescence-based biosensors

    DEFF Research Database (Denmark)

    Rybtke, Morten; Chua, Song Lin; Yam, Joey Kuok Hoong

    2017-01-01

    developed a collection of fluorescence-based c-di-GMP biosensors capable of gauging the c-di-GMP level in Pseudomonas aeruginosa and closely related bacteria. Here, we describe protocols for the use of these biosensors in gauging and visualizing cellular c-di-GMP levels of P. aeruginosa both in in vitro...

  4. Fluorescent ester dye-based assays for the in vitro measurement of Neospora caninum proliferation.

    Science.gov (United States)

    Mota, Caroline M; Ferreira, Marcela D; Costa, Lourenço F; Barros, Patrício S C; Silva, Murilo V; Santiago, Fernanda M; Mineo, José R; Mineo, Tiago W P

    2014-09-15

    Techniques for the measurement of parasite loads in different experimental models have evolved throughout the years. The quantification of stained slides using regular cytological stains is currently the most common technique. However, this modality of evaluation is labor-intensive, and the interpretation of the results is subjective because the successes of the assays mainly rely on the abilities of the professionals involved. Moreover, the novel genetic manipulation techniques that are commonly applied for closely related Toxoplasma gondii have not yet been developed for Neospora caninum. Thus, we aimed to develop a simple protocol for parasite quantification using pre-stained N. caninum tachyzoites and fluorescent probes based on ester compounds (i.e., CFSE and DDAO). For this purpose, we employed a quantification procedure based on flow cytometry analysis. Pre-stained parasites were also examined with a fluorescent microscope, which revealed that both dyes were detectable. Direct comparison of the numbers of CFSE+ and DDAO+ cells to the values obtained with classical cytology techniques yielded statistically comparable results that also accorded with genomic DNA amplification results. Although the fluorescence emitted by DDAO was more intense and provided better discrimination between the populations of parasitized cells, CFSE+ tachyzoites were detected for several days. In conclusion, this study describes a simple, fast, low-cost and reproducible protocol for N. caninum quantification that is based on parasite pre-staining with fluorescent ester-based probes. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy

    NARCIS (Netherlands)

    Singh, A.P.; Krieger, J.W.; Buchholz, J.; Charbon, E.; Langowski, J.; Wohland, T.

    2013-01-01

    Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, transport, flow velocities and concentrations in an imaging mode. SPIM-FCS records correlation functions over a whole plane in

  6. A Fluorescence-Based Assay for Identification of Bacterial Topoisomerase I Poisons.

    Science.gov (United States)

    Annamalai, Thirunavukkarasu; Cheng, Bokun; Keswani, Neelam; Tse-Dinh, Yuk-Ching

    2018-01-01

    Bacterial Topoisomerase I is a potential target for the identification of novel topoisomerase poison inhibitors that could provide leads for a new class of antibacterial compounds. Here we describe in detail a fluorescence-based cleavage assay that is successfully used in HTS for the discovery of bacterial topoisomerase Ι poisons.

  7. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE: INDUCED BY RADIATION, CHEMICALS AND ENZYMES

    Science.gov (United States)

    A simple and rapid assay to detect DNA damage is reported. This assay is based on the ability of certain dyes to fluoresce upon intercalation with dsDNA. Damage caused by ultraviolet (UV) radiation, chemicals or restriction enzymes is detected using this assay. UV radiation at...

  8. Water soluble chromone Schiff base derivatives as fluorescence receptor for aluminium(III)

    Czech Academy of Sciences Publication Activity Database

    Jakubek, M.; Kejík, Z.; Parchaňský, Václav; Kaplánek, R.; Vasina, L.; Martásek, P.; Král, V.

    2017-01-01

    Roč. 29, č. 1 (2017), s. 1-7 ISSN 1061-0278 R&D Projects: GA TA ČR(CZ) TE01020028 Institutional support: RVO:61388963 Keywords : aluminium sensing * chelator * chromone * fluorescence * Schiff base Subject RIV: CF - Physical ; Theoretical Chemistry OBOR OECD: Physical chemistry Impact factor: 1.264, year: 2016

  9. Fluorescent BINOL-based sensor for thorium recognition and a density functional theory investigation

    International Nuclear Information System (INIS)

    Wen, Jun; Dong, Liang; Tian, Jie; Jiang, Tao; Yang, Yan-Qiu; Huang, Zeng; Yu, Xiao-Qi; Hu, Chang-Wei; Hu, Sheng; Yang, Tong-Zai; Wang, Xiao-Lin

    2013-01-01

    Graphical abstract: A novel BINOL fluorescence sensor L-1 for the recognition of thorium ion with high selectivity and sensitivity. -- Highlights: • The first case of one-to-one stoichiometric responding fluorescent sensor for thorium. • An easy preparation and novel BINOL-based chemical sensor. • This sensor for thorium ion recognition by fluorescence spectrophotometry with high selectivity and sensitivity. • This sensor shows good accuracy for analysis of thorium ions in river water. • DFT calculations indicate that a strong binding interaction exists between the L-1 and Th 4+ . -- Abstract: A novel 1,1′-bi-2-naphthol (BINOL) derivative fluorescence sensor L-1 for the recognition of thorium ion with a fluorescence quench response. This ligand showed high selectivity and sensitivity for thorium ion recognition. Coordination effects were investigated by DFT calculations, and the coordination modes and sites were confirmed. Moreover, the coordination abilities of the L-1 ligand with Th 4+ and UO 2 2+ were evaluated

  10. The sensitivity and selectivity properties of a fluorescence sensor based on quinoline-Bodipy

    Energy Technology Data Exchange (ETDEWEB)

    Nuri Kursunlu, Ahmed, E-mail: ankursunlu@gmail.com; Guler, Ersin

    2014-01-15

    A novel florescence sensor (Q-BODIPY) based on quinoline-Bodipy (quinoline-boradiazaindacene) was prepared by ‘click chemistry’ in several stages. The sensing actions of Q-BODIPY were confirmed by UV–vis titration, emission and excitation spectroscopic studies in presence of Mn{sup 2+}, Co{sup 2+}, Ni{sup 2+}, Cu{sup 2+}, Zn{sup 2+}, Cd{sup 2+}, Sn{sup 2+}, Hg{sup 2+}, Pb{sup 2+}, La{sup 3+}, Ga{sup 3+}, Er{sup 3+} and Yb{sup 3+} ions in methanol:H{sub 2}O (1:1) medium. Whereas some metal ions can only cause quenching effect on the fluorescence intensity of Q-BODIPY, some of them show an increase in fluorescence intensity. The stoichiometry of host–guest complexes formed was determined by Job′s plot method. The binding constants were calculated by Stern–Volmer method. As a fluorescence sensor, Q-BODIPY shows the best selectivity performance against Zn{sup 2+} ions in according to all spectroscopic data. -- Highlights: • Q-BODIPY prepared by several techniques shows a fluorescent behavior toward p, d and f block metal ions. • Q-BODIPY has both a more sensitivity and more effective ability for the detection of Zn(II) ion. • The synthesis strategies to produce Bodipy′s with metal coordinating offer a new approach for the design of novel fluorescence sensors.

  11. Detection of DNA hybridization based on SnO2 nanomaterial enhanced fluorescence

    International Nuclear Information System (INIS)

    Gu Cuiping; Huang Jiarui; Ni Ning; Li Minqiang; Liu Jinhuai

    2008-01-01

    In this paper, enhanced fluorescence emissions were firstly investigated based on SnO 2 nanomaterial, and its application in the detection of DNA hybridization was also demonstrated. The microarray of SnO 2 nanomaterial was fabricated by the vapour phase transport method catalyzed by patterned Au nanoparticles on a silicon substrate. A probe DNA was immobilized on the substrate with patterned SnO 2 nanomaterial, respectively, by covalent and non-covalent linking schemes. When a fluorophore labelled target DNA was hybridized with a probe DNA on the substrate, fluorescence emissions were only observed on the surface of SnO 2 nanomaterial, which indicated the property of enhancing fluorescence signals from the SnO 2 nanomaterial. By comparing the different fluorescence images from covalent and non-covalent linking schemes, the covalent method was confirmed to be more effective for immobilizing a probe DNA. With the combined use of SnO 2 nanomaterial and the covalent linking scheme, the target DNA could be detected at a very low concentration of 10 fM. And the stability of SnO 2 nanomaterial under the experimental conditions was also compared with silicon nanowires. The findings strongly suggested that SnO 2 nanomaterial could be extensively applied in detections of biological samples with enhancing fluorescence property and high stability

  12. Color-Tunable Solid-State Fluorescence Emission from Carbazole-Based BODIPYs.

    Science.gov (United States)

    Maeda, Chihiro; Todaka, Takumi; Ueda, Tomomi; Ema, Tadashi

    2016-05-23

    Several carbazole-based boron dipyrromethene (BODIPY) dyes were synthesized by organometallic approaches. Thiazole, benzothiazole, imidazole, benzimidazole, triazole, and indolone substituents were introduced at the 1-position of the carbazole moiety, and boron complexation of each dipyrrin generated the corresponding compounds 1, 2 a, and 3-6. The properties of these products were investigated by UV/Vis and fluorescence spectroscopy, cyclic voltammetry, X-ray crystallography, and DFT calculations. These compounds exhibited large Stokes shifts, and compounds 1, 2 a, and 3-5 fluoresced both in solution and in the solid state. Complex 2 a showed the highest fluorescence quantum yield (ΦF ) in the solid state, therefore boron complexes of the carbazole-benzothiazole hybrids 2 b-f, which had several different substituents, were prepared and the effects of the substituents on the photophysical properties of the compounds were examined. The fluorescence properties showed good correlation with the results of crystal-packing analyses, and the dyes exhibited color-tunable solid-state fluorescence. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. A coumarin-based colorimetric fluorescent probe for hydrogen sulfide

    Indian Academy of Sciences (India)

    shift, good solubility and high quantum yield.10 Previ- ously, we reported a coumarin-based chromogenic and ratiometric probe for hydrazine.11 As we know, when the phenolic hydroxyl of a fluorophore is protected via esterification, no emission of the fluorophore would be found due to the electron-withdrawing property of.

  14. Synthesis and characterisation of highly fluorescent core–shell nanoparticles based on Alexa dyes

    International Nuclear Information System (INIS)

    Natte, Kishore; Behnke, Thomas; Orts-Gil, Guillermo; Würth, Christian; Friedrich, Jörg F.; Österle, Werner; Resch-Genger, Ute

    2012-01-01

    Current and future developments in the emerging field of nanobiotechnology are closely linked to the rational design of novel fluorescent nanomaterials, e.g. for biosensing and imaging applications. Here, the synthesis of bright near infrared (NIR)-emissive nanoparticles based on the grafting of silica nanoparticles (SNPs) with 3-aminopropyl triethoxysilane (APTES) followed by covalent attachment of Alexa dyes and their subsequent shielding by an additional silica shell are presented. These nanoparticles were investigated by dynamic light scattering (DLS), transmission electron microscopy (TEM) and fluorescence spectroscopy. TEM studies revealed the monodispersity of the initially prepared and fluorophore-labelled silica particles and the subsequent formation of raspberry-like structures after addition of a silica precursor. Measurements of absolute fluorescence quantum yields of these scattering particle suspensions with an integrating sphere setup demonstrated the influence of dye labelling density-dependent fluorophore aggregation on the signaling behaviour of such nanoparticles.

  15. Fluorescence Probe Based on an Amino-Functionalized Fluorescent Magnetic Nanocomposite for Detection of Folic Acid in Serum.

    Science.gov (United States)

    Li, Xiaowan; Chen, Ligang

    2016-11-23

    A new fluorescence probe constructed with a multifunctional nanocomposite, Fe 3 O 4 -ZnS:Mn 2+ /SiO 2 -NH 2 , was successfully synthesized and then used to detect folic acid in real serum samples. The nanocomposite was characterized by fluorescence spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray powder diffraction, and physical property measurement system. With the addition of analyte, the Fe 3 O 4 -ZnS:Mn 2+ /SiO 2 -NH 2 composite and folic acid formed a new complex because cross-linking of the amino and carboxyl groups participated in the condensation reaction. Then, the energy of quantum dots was transferred to the complex and led to quenching of the fluorescence. Moreover, the fluorescence intensity decreased significantly as the concentration of folic acid increased, and the fluorescence quenching ratio F 0 /F was related to the folic acid concentration in the range from 0.1 to 5 μg mL -1 . This method was used for detecting folic acid in real serum samples and gave recoveries in the range of 89.0%-96.0%, with relative standard deviations of 1.2%-3.9%. The detection limit was 9.6 ng mL -1 (S/N = 3). These satisfactory and simple results showed the great potential of this fluorescence probe in the field of pharmaceutical analysis.

  16. Exploiting multimode waveguides for pure fibre based fluorescence imaging

    Science.gov (United States)

    Čižmár, TomáÅ.¡; Dholakia, Kishan

    2013-03-01

    There has been an immense drive in modern microscopy towards miniaturisation and _bre based technology. This has been necessitated by the need to access hostile or difficult environments particulalrly in-situ and in-vivo. Strategies to date have included the use of specialist fibres and miniaturised scanning systems accompanied by ingenious microfabricated lenses. In parallel recent studies of randomized light fields and their holographic control opened up new ways for imaging. We present a novel approach for this field by utilising disordered light within a standard multimode optical fibre for minimally invasive lenseless microscopy and optical mode conversion. We demonstrate scanning uorescence microscopy at acquisition rates allowing observation of dynamic processes such as Brownian motion of mesoscopic particles. As the sample plane can be defined at any distance from the fibre facet, we eliminate the need for complex or elaborate focusing optics (e.g. miniaturized objectives, GRIN lenses) and instead reconfigure the system dynamically to image different axial planes. Furthermore, we show how such control can realise a new form of mode converter and generate various types of advanced light fields such as propagation-invariant beams and optical vortices. These may be useful for future fibre based implementations of super-resolution or light sheet microscopy. To the best of our knowledge, this technology represents the narrowest possible image guiding system based on light propagation.

  17. A fluorescent sensor based on binaphthol-quinoline Schiff base for ...

    Indian Academy of Sciences (India)

    moiety to quinoline fluorophore and the isomerization process of C=N bond.12 Upon addition of 4.0 equiv. of. Zn2+, a dramatic fluorescence enhancement at 512nm was observed, which could be attributed to the Zn2+ - binding induced suppression of PET and C=N isome rization. The Zn2+ induced green fluorescence ...

  18. A recombinant fusion protein-based, fluorescent protease assay for high throughput-compatible substrate screening.

    Science.gov (United States)

    Bozóki, Beáta; Gazda, Lívia; Tóth, Ferenc; Miczi, Márió; Mótyán, János András; Tőzsér, József

    2018-01-01

    In connection with the intensive investigation of proteases, several methods have been developed for analysis of the substrate specificity. Due to the great number of proteases and the expected target molecules to be analyzed, time- and cost-efficient high-throughput screening (HTS) methods are preferred. Here we describe the development and application of a separation-based HTS-compatible fluorescent protease assay, which is based on the use of recombinant fusion proteins as substrates of proteases. The protein substrates used in this assay consists of N-terminal (hexahistidine and maltose binding protein) fusion tags, cleavage sequences of the tobacco etch virus (TEV) and HIV-1 proteases, and a C-terminal fluorescent protein (mApple or mTurquoise2). The assay is based on the fluorimetric detection of the fluorescent proteins, which are released from the magnetic bead-attached substrates by the proteolytic cleavage. The protease assay has been applied for activity measurements of TEV and HIV-1 proteases to test the suitability of the system for enzyme kinetic measurements, inhibition studies, and determination of pH optimum. We also found that denatured fluorescent proteins can be renatured after SDS-PAGE of denaturing conditions, but showed differences in their renaturation abilities. After in-gel renaturation both substrates and cleavage products can be identified by in-gel UV detection. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Factors affecting use of fluorescent agents in identification of resin-based polymers.

    Science.gov (United States)

    D'Alpino, Paulo H P; Pereira, José C; Svizero, Nádia R; Rueggeberg, Frederick A; Pashley, David H

    2006-10-01

    The present study is a compilation of methodologies developed in order to evaluate the effects of addition of a fluorescent agent, rhodamine B, to resin-based materials. The intent of the work was to develop a systematic methodology that accounts for variables not shown to be of concern in past testing, but may significantly affect interpretation of the resulting images and material properties. Different methodologies were specifically developed to evaluate factors affecting the use of the fluorescent agent rhodamine B in the identification of resin-based materials. The influence of dye concentration was evaluated with respect to adhesive polymerization reaction kinetics and bond strength of the resin into which the dye is added. The pH of the dye-solvent solution was also considered. Additionally, the mass and percentage of rhodamine B leached into different media over time was determined. It was demonstrated that a specific dye must be used with caution, and the concentration of a fluorescent agent is important with respect to both the conversion and bond strength of the resin to which the dye is added. An alcohol-based dye solution also produced an extremely high amount of dye leaching from polymerized specimens. The adhesive pH was not altered. The standardized methodology developed here to analyze restoration interfaces using a fluorescent dye can lessen the impact of variables when interpreting results.

  20. Molecularly Imprinted Polymer/Metal Organic Framework Based Chemical Sensors

    Directory of Open Access Journals (Sweden)

    Zhenzhong Guo

    2016-10-01

    Full Text Available The present review describes recent advances in the concept of molecular imprinting using metal organic frameworks (MOF for development of chemical sensors. Two main strategies regarding the fabrication, performance and applications of recent sensors based on molecularly imprinted polymers associated with MOF are presented: molecularly imprinted MOF films and molecularly imprinted core-shell nanoparticles using MOF as core. The associated transduction modes are also discussed. A brief conclusion and future expectations are described herein.

  1. Hybrid Imaging Labels: Providing the Link Between Mass Spectrometry-Based Molecular Pathology and Theranostics

    Science.gov (United States)

    Buckle, Tessa; van der Wal, Steffen; van Malderen, Stijn J.M.; Müller, Larissa; Kuil, Joeri; van Unen, Vincent; Peters, Ruud J.B.; van Bemmel, Margaretha E.M.; McDonnell, Liam A.; Velders, Aldrik H.; Koning, Frits; Vanhaeke, Frank; van Leeuwen, Fijs W. B.

    2017-01-01

    Background: Development of theranostic concepts that include inductively coupled plasma mass spectrometry (ICP-MS) and laser ablation ICP-MS (LA-ICP-MS) imaging can be hindered by the lack of a direct comparison to more standardly used methods for in vitro and in vivo evaluation; e.g. fluorescence or nuclear medicine. In this study a bimodal (or rather, hybrid) tracer that contains both a fluorescent dye and a chelate was used to evaluate the existence of a direct link between mass spectrometry (MS) and in vitro and in vivo molecular imaging findings using fluorescence and radioisotopes. At the same time, the hybrid label was used to determine whether the use of a single isotope label would allow for MS-based diagnostics. Methods: A hybrid label that contained both a DTPA chelate (that was coordinated with either 165Ho or 111In) and a Cy5 fluorescent dye was coupled to the chemokine receptor 4 (CXCR4) targeting peptide Ac-TZ14011 (hybrid-Cy5-Ac-TZ4011). This receptor targeting tracer was used to 1) validate the efficacy of (165Ho-based) mass-cytometry in determining the receptor affinity via comparison with fluorescence-based flow cytometry (Cy5), 2) evaluate the microscopic binding pattern of the tracer in tumor cells using both fluorescence confocal imaging (Cy5) and LA-ICP-MS-imaging (165Ho), 3) compare in vivo biodistribution patterns obtained with ICP-MS (165Ho) and radiodetection (111In) after intravenous administration of hybrid-Cy5-Ac-TZ4011 in tumor-bearing mice. Finally, LA-ICP-MS-imaging (165Ho) was linked to fluorescence-based analysis of excised tissue samples (Cy5). Results: Analysis with both mass-cytometry and flow cytometry revealed a similar receptor affinity, respectively 352 ± 141 nM and 245 ± 65 nM (p = 0.08), but with a much lower detection sensitivity for the first modality. In vitro LA-ICP-MS imaging (165Ho) enabled clear discrimination between CXCR4 positive and negative cells, but fluorescence microscopy was required to determine the

  2. Winter wheat GPC estimation with fluorescence-based sensor measurements of canopy

    Science.gov (United States)

    Song, Xiaoyu; Wang, Jihua; Gu, Xiaohe; Xu, Xingang

    2015-10-01

    This study focused on the wheat grain protein content (GPC) estimation based on wheat canopy chlorophyll parameters which acquired by hand-held instrument, Multiplex 3. Nine fluorescence spectral indices from Multiplex sensor were used in this study. The wheat GPC estimation experiment was conducted in 2012 at the National Experiment Station for Precision Agriculture in Changping district, Beijing. A square with area of 1.1 ha was selected and divided to 110 small plots by 10×10m in this study. In each plot, four 1-m2 area distributed in the square were selected for canopy fluorescence spectral measurements, physiological and biochemical analyses. Measurements were performed five times at wheat raising, jointing, heading stage, milking and ripening stage, respectively. The wheat plant samples for each plot were then collected after the measurement and sent to Lab for leaf N concentration (LNC) and canopy nitrogen density (CND) analyzed. GPC sampling for each plot was collected manually during the harvested season. Then, statistical analysis were performed to detect the correlation between fluorescence spectral indices and wheat CND for each growth stage, as well as GPC. The results indicate that two Nitrogen Balance Indices, NBI_G and NBI_R were more sensitive to wheat GPC than other fluorescence spectral indices at milking stage and ripening stage. Five linear regression models with GPC and fluorescence indices at different winter wheat growth stages were then established. The R2 of GPC estimated model increased form 0.312 at raising stage to 0.686 at ripening stage. The study reveals that canopy-level fluorescence spectral parameters were better indicators for the wheat group activity and could be demonstrated to be good indicators for winter wheat GPC estimation.

  3. Determination of Europium by fluorescence using pyrrolidinium based task specific ionic liquid

    International Nuclear Information System (INIS)

    Kumar, Satendra; Gayan, Siuli Maji; Sankaran, K.

    2012-01-01

    Determination of lanthanides using fluorescence is a major challenge in aqueous medium due to their poor molar absorptivities and low quantum yield. To overcome these problems, ligand sensitized fluorescence has been widely used for trace level detection of lanthanides in solutions. Recently, ionic liquid a non aqueous medium has been used to observe the fluorescence of lanthanides. In this work we have used pyrrolidinium based ionic liquid for the study of europium fluorescence. The ionic liquid (bmpyr)(BA) was prepared using the metathesis reaction involving 1-butyl,1-methyl pyrrolidinium chloride (bmpyr)(CI). (bmpyr)(Cl) crystals and silver benzoate were added according to their mole equivalents in methanol and stirred for 4 hrs. The filtrate was concentrated using a rotary evaporator, dried for 10 h at room temperature, and then for 40 h at 105℃under reduced pressure. This results in a white crystalline hygroscopic solid which was characterized by FT-lR spectroscopy. The yield of the product was ∼ 95%. Butyl-1, methyl pyrrolidinium bis(trifluoromethanesulfonyl)amide (bmpyr)(TF 2 N) is used for dissolve and dilute (bmpyr)(BA) and fluorescence and life time of Eu 3+ were obtained. For the concentration of Eu 3+ used in this study (2x10 -7 M), the optimum concentration of the sensitizing ionic liquid was found to be 10 -4 M. An enhancement factor of about 32000 was found in this ionic liquid compare to aqueous medium. Fluorescence life time of europium in this ionic liquid is 1100 μs which is ten times more than the life time of europium in aqueous medium (110 μs), indicating a reduction in the rates of non-radiative processes which was provided by the ionic liquid. This study leads to detection of europium in ppb level. (author)

  4. Fluorescent Biosensor for Phosphate Determination Based on Immobilized Polyfluorene-Liposomal Nanoparticles Coupled with Alkaline Phosphatase.

    Science.gov (United States)

    Kahveci, Zehra; Martínez-Tomé, Maria José; Mallavia, Ricardo; Mateo, C Reyes

    2017-01-11

    This work describes the development of a novel fluorescent biosensor based on the inhibition of alkaline phosphatase (ALP). The biosensor is composed of the enzyme ALP and the conjugated cationic polyfluorene HTMA-PFP. The working principle of the biosensor is based on the fluorescence quenching of this polyelectrolyte by p-nitrophenol (PNP), a product of the hydrolysis reaction of p-nitrophenyl phosphate (PNPP) catalyzed by ALP. Because HTMA-PFP forms unstable aggregates in buffer, with low fluorescence efficiency, previous stabilization of the polyelectrolyte was required before the development of the biosensor. HTMA-PFP was stabilized through its interaction with lipid vesicles to obtain stable blue-emitting nanoparticles (NPs). Fluorescent NPs were characterized, and the ability to be quenched by PNP was evaluated. These nanoparticles were coupled to ALP and entrapped in a sol-gel matrix to produce a biosensor that can serve as a screening platform to identify ALP inhibitors. The components of the biosensor were examined before and after sol-gel entrapment, and the biosensor was optimized to allow the determination of phosphate ion in aqueous medium.

  5. Fluorescence-based determination of the copper concentration in drinking water

    Science.gov (United States)

    Hötzer, Benjamin; Scheu, Timo; Jung, Gregor; Castritius, Stefan

    2013-05-01

    Copper is a heavy metal, which is used in heat and electrical conductors and in a multitude of alloys in the technical context. Moreover, it is a trace element that is essential for the life of organisms but can cause toxic effects in elevated concentrations. Maximum limits in water and beverages exist. Here, the decrease of the fluorescence lifetime of green fluorescent protein (GFP) by Förster resonance energy transfer is used to measure the copper ion concentration in drinking water. Therefore, a system is developed that is based on a GFP sample in a predefined concentration. The GFP mutant can be excited with blue light. For binding of copper ions, a His-tag is included in the GFP. After measuring the fluorescence lifetime of pure GFP, the copper determination of the sample is performed by lifetime measurement. Therefore, the lifetime can be assigned to the copper concentration of the GFP-doped drinking water sample. In summary, a method for the quantification of copper ions based on changes of the fluorescence lifetime of GFP is developed, and the measurement of the copper concentration in water samples is performed.

  6. Image overlay solution based on threshold detection for a compact near infrared fluorescence goggle system.

    Science.gov (United States)

    Gao, Shengkui; Mondal, Suman B; Zhu, Nan; Liang, RongGuang; Achilefu, Samuel; Gruev, Viktor

    2015-01-01

    Near infrared (NIR) fluorescence imaging has shown great potential for various clinical procedures, including intraoperative image guidance. However, existing NIR fluorescence imaging systems either have a large footprint or are handheld, which limits their usage in intraoperative applications. We present a compact NIR fluorescence imaging system (NFIS) with an image overlay solution based on threshold detection, which can be easily integrated with a goggle display system for intraoperative guidance. The proposed NFIS achieves compactness, light weight, hands-free operation, high-precision superimposition, and a real-time frame rate. In addition, the miniature and ultra-lightweight light-emitting diode tracking pod is easy to incorporate with NIR fluorescence imaging. Based on experimental evaluation, the proposed NFIS solution has a lower detection limit of 25 nM of indocyanine green at 27 fps and realizes a highly precise image overlay of NIR and visible images of mice in vivo. The overlay error is limited within a 2-mm scale at a 65-cm working distance, which is highly reliable for clinical study and surgical use.

  7. A New Quinoline-Based Acylhydrazone for Highly Selective Fluorescence Recognition of Cu(II) and Sulfide in Aqueous Solution

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Lijun; Zhou, Pei; Huang, Zhenlong; Zhao, Jia; Cai, Mingjun [Bohai Univ., Jinzhou (China); Qi, Zhikai [Shanxi Normal Univ., Linfen (China)

    2013-08-15

    A new quinoline-based acylhydrazone (1) has been synthesized and applied as a fluorescent probe. Probe 1 exhibits high selectivity and sensitivity to Cu{sup 2+} with fluorescence 'ON-OFF' behavior in HEPES buffered (1% DMSO, HEPES 20 mM, pH = 7.4) solution. The on-site generated 1-Cu{sup 2+} complex displays excellent selectivity to sulfide ions with fluorescence 'OFF-ON' performance through copper displacement approach.

  8. Rapid molecular cytogenetic analysis of X-chromosomal microdeletions: Fluorescence in situ hybridization (FISH) for complex glycerol kinase deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Worley, K.C.; Lindsay, E.A.; McCabe, E.R.B. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1995-07-17

    Diagnosis of X-chromosomal microdeletions has relied upon the traditional methods of Southern blotting and DNA amplification, with carrier identification requiring time-consuming and unreliable dosage calculations. In this report, we describe rapid molecular cytogenetic identification of deleted DNA in affected males with the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency, CGKD) and female carriers for this disorder. CGKD deletions involve the genes for glycerol kinase, Duchenne muscular dystrophy, and/or adrenal hypoplasia congenita. We report an improved method for diagnosis of deletions in individuals with CGKD and for identification of female carriers within their families using fluorescence in situ hybridization (FISH) with a cosmid marker (cosmid 35) within the glycerol kinase gene. When used in combination with an Xq control probe, affected males demonstrate a single signal from the control probe, while female carriers demonstrate a normal chromosome with two signals, as well as a deleted chromosome with a single signal from the control probe. FISH analysis for CGKD provides the advantages of speed and accuracy for evaluation of submicroscopic X-chromosome deletions, particularly in identification of female carriers. In addition to improving carrier evaluation, FISH will make prenatal diagnosis of CGKD more readily available. 17 refs., 2 figs.

  9. A fluorescence chemosensor based on peptidase for detecting nickel(II) with high selectivity and high sensitivity.

    Science.gov (United States)

    Lv, Xiao-Li; Luo, Shi-Zhong

    2012-03-01

    We report herein a new class of metal ion chemosensors and give the first example of a metal-dependent peptidase chemosensor for metal ions. The chemosensor contains the basic specific Ni(II)-dependent peptide bond hydrolysis sequence (Gly-Ala-Ser-Arg-His-Trp-Lys-Phe-Lys). The substrate was labeled with a fluorophore at the N-terminal and a quencher at the C-terminal Lys side chain. Initially, the MOCAc ((7-methoxycoumarin-4-yl)acetyl-) emission was quenched by the nearby quencher. In the presence of Ni(II), the substrate was irreversibly cleaved at the cleavage site, leading to a 20-fold increase in fluorescence intensity. The chemosensor combines the high selectivity of a peptidase (at least greater than tenfold for Ni(II) over other metal ions) with the high sensitivity of fluorescence detection limit of 50 nM and can be applied for the quantitative detection of Ni(II) over a concentration range of three orders of magnitude. Given this degree of selectivity and sensitivity, our molecular engineering design may prove useful in the future development of other peptidase-based probes for different metal ions in toxicological and environmental monitoring.

  10. The N-B Interaction through a Water Bridge: Understanding the Chemoselectivity of a Fluorescent Protein Based Probe for Peroxynitrite.

    Science.gov (United States)

    Chen, Zhi-jie; Tian, Ziqi; Kallio, Karen; Oleson, April L; Ji, Ao; Borchardt, Dan; Jiang, De-en; Remington, S James; Ai, Hui-wang

    2016-04-13

    Boronic acid and esters have been extensively utilized for molecular recognition and chemical sensing. We recently reported a genetically encoded peroxynitrite (ONOO(-))-specific fluorescent sensor, pnGFP, based on the incorporation of a boronic acid moiety into a circularly permuted green fluorescent protein (cpGFP) followed by directed protein evolution. Different from typical arylboronic acids and esters, the chromophore of pnGFP is unreactive to millimolar concentrations of hydrogen peroxide (H2O2). The focus of this study is to explore the mechanism for the observed unusual chemoselectivity of pnGFP toward peroxynitrite over hydrogen peroxide by using site-directed mutagenesis, X-ray crystallography, (11)B NMR, and computational analysis. Our data collectively support that a His residue on the protein scaffold polarizes a water molecule to induce the formation of an sp(3)-hybridized boron in the chromophore, thereby tuning the reactivity of pnGFP with various reactive oxygen and nitrogen species (ROS/RNS). Our study demonstrates the first example of tunable boron chemistry in a folded nonnative protein, which offers wide implications in designing selective chemical probes.

  11. The Fluorescence Enhancement of Mercury Detected in Food Based on Rhodamine Derivatives.

    Science.gov (United States)

    Fan, Cai-ling; Xie, Pu-hui; Cui, Shu-min; Yang, Li-na; Sun, Qing; Ai, Zhi-lu

    2015-05-01

    Recently, the problem of food security is more and more serious, and people pay attention to mercury because of the toxic of it. A new approach for the determination of mercury content in foodstuff is devised. In this paper, first, we design and synthesis a new kind of fluorescent probe whose matrix based on rhodamine B, hydrazine hydrate and hydroxy benzaldehyde. Through the analysis of H-NMR spectra of the synthesized product L1, we confirm that the synthetic substance is the adjacent carboxyl benzaldehyde hydrazone structure generation of rhodamine B. Then, we measure the fluorescence signal intensity of the probe with different concentrations of mercury ions fully upon complexation by fluorescence spectrometer and we can study the relationship between the mercury ion concentration and the fluorescence intensity and draw the standard working curve. Following, It's time to discuss the microwave digestion processing of tea, after digestion we use the synthetic probe Li for determination of mercury content in tea. The experimental results show that the maximum excitation wavelength of the probe and coordination compound are 568. 05 and 560. 00 nm, the maximum emission wavelength are 587. 94 and 580. 00 nm. Then we can find the best testing conditions to improve the degree of accuracy, that is: room temperature, 50% the methanol solution, 3. 0 mL pH 4. 0 buffer solution, in the extent of 30 min. The experimental results show that Na+, K+, Ca2+, Cu2+, Zn2+, Al3+ have little impact on the fluorescence intensity of the:probe. Fe3+, Mg2+, Ba2+ has a weak enhancement to the fluorescence intensity of the probe. While a low concentrations of Hg2+ have an obviously enhanced effect on the fluorescence intensity of the probe. In contrast to other metal ions, the probe for Hg2+ has a good selectivity. Linear relationship between the magnitude of increase in fluorescence intensity and concentration of mercury ion was in the range of 5~20 ng . L-1 with detection limit (3S/N) of

  12. Hybrid Microfluidic Platform for Multifactorial Analysis Based on Electrical Impedance, Refractometry, Optical Absorption and Fluorescence

    Directory of Open Access Journals (Sweden)

    Fábio M. Pereira

    2016-10-01

    Full Text Available This paper describes the development of a novel microfluidic platform for multifactorial analysis integrating four label-free detection methods: electrical impedance, refractometry, optical absorption and fluorescence. We present the rationale for the design and the details of the microfabrication of this multifactorial hybrid microfluidic chip. The structure of the platform consists of a three-dimensionally patterned polydimethylsiloxane top part attached to a bottom SU-8 epoxy-based negative photoresist part, where microelectrodes and optical fibers are incorporated to enable impedance and optical analysis. As a proof of concept, the chip functions have been tested and explored, enabling a diversity of applications: (i impedance-based identification of the size of micro beads, as well as counting and distinguishing of erythrocytes by their volume or membrane properties; (ii simultaneous determination of the refractive index and optical absorption properties of solutions; and (iii fluorescence-based bead counting.

  13. Light extraction efficiency enhancement for fluorescent SiC based white light-emitting diodes

    DEFF Research Database (Denmark)

    Ou, Haiyan; Ou, Yiyu; Argyraki, Aikaterini

    Fluorescent SiC based white light-emitting diodes(LEDs) light source, as an innovative energy-efficient light source, would even have longer lifetime, better light quality and eliminated blue-tone effect, compared to the current phosphor based white LED light source. In this paper, the yellow....... At a device level, the focus is on improving the light extraction efficiency due to the rather high refractive index of SiC by nanostructuring the surface of SiC. Both periodic nanostructures made by e-beam lithography and nanosphere lithography and random nanostructures made by self-assembled Au nanosphere...... mask and a thin layer of Al film have been investigated and all of them showed much enhanced extraction efficiency. All these good results pave the way to a very promising fluorescent SiC based white LED light source...

  14. Investigation of molecular mechanisms of action of chelating drugs on protein-lipid model membranes by X-ray fluorescence

    International Nuclear Information System (INIS)

    Novikova, N. N.; Zheludeva, S. I.; Koval'chuk, M. V.; Stepina, N. D.; Erko, A. I.; Yur'eva, E. A.

    2009-01-01

    Protein-lipid films based on the enzyme alkaline phosphatase were subjected to the action of chelating drugs, which are used for accelerating the removal of heavy metals from the human body, and the elemental composition of the resulting films was investigated. Total-reflection X-ray fluorescence measurements were performed at the Berlin Electron Storage Ring Company for Synchrotron Radiation (BESSY) in Germany. A comparative estimation of the protective effect of four drugs (EDTA, succimer, xydiphone, and mediphon) on membrane-bound enzymes damaged by lead ions was made. The changes in the elemental composition of the protein-lipid films caused by high doses of chelating drugs were investigated. It was shown that state-of-the-art X-ray techniques can, in principle, be used to develop new methods for the in vitro evaluation of the efficiency of drugs, providing differential data on their actions.

  15. Measurements of molecular carbon radical concentrations by saturated laser-induced fluorescence in hydrocarbon flames at atmospheric pressure

    Science.gov (United States)

    Marrocco, Michele; D'Apice, Massimo; Giammartini, Stefano; Magaldi, M.; Romano, G. P.

    2003-09-01

    The C2 abundance in flames at atmospheric pressure is reported for three fuel-rich combustion environments: acetylene/oxygen, Liquid Petroleum Gas (LPG)/air and methane/air flames. Measurements have been based on spectrographic and two-dimensional Laser-Induced Fluorescence (LIF) of the Δv = -1 d3IIg-a3IIu band. The oxyacetylene flame was utilized to establish a comparison with literature data and absolute values, calibrated through Rayleigh scattering, were found to be distributed within the 1014 cm-3 range. The induced emission from two Bunsen flames fed with LPG/air or methane/air was also investigated. Accumulated acquisitions were necessary to detect small C2 quantities in the order of 1010 cm-3 concentrated on the flame front.

  16. Cyclin B1 Destruction Box-Mediated Protein Instability: The Enhanced Sensitivity of Fluorescent-Protein-Based Reporter Gene System

    Directory of Open Access Journals (Sweden)

    Chao-Hsun Yang

    2013-01-01

    Full Text Available The periodic expression and destruction of several cyclins are the most important steps for the exact regulation of cell cycle. Cyclins are degraded by the ubiquitin-proteasome system during cell cycle. Besides, a short sequence near the N-terminal of cyclin B called the destruction box (D-box; CDB is also required. Fluorescent-protein-based reporter gene system is insensitive to analysis because of the overly stable fluorescent proteins. Therefore, in this study, we use human CDB fused with both enhanced green fluorescent protein (EGFP at C-terminus and red fluorescent protein (RFP, DsRed at N-terminus in the transfected human melanoma cells to examine the effects of CDB on different fluorescent proteins. Our results indicated that CDB-fused fluorescent protein can be used to examine the slight gene regulations in the reporter gene system and have the potential to be the system for screening of functional compounds in the future.

  17. Optical fiber-based photomechanical molecular delivery system

    Science.gov (United States)

    Nakano, Koki; Sato, Shunichi; Kawauchi, Satoko; Ashida, Hiroshi; Nishidate, Izumi

    2014-02-01

    Molecular delivery based on nanosecond pulsed laser-induced photomechanical waves (PMWs) enables endoscopic application by using an optical fiber for laser transmission. In our previous fiber system, a laser target, which was a black natural rubber film as a laser absorbing material covered with an optically transparent polyethylene terephthalate disk to confine the laser-induced plasma, was attached to the output end of a 1 mm core diameter quartz fiber. There were two problems in that system: 1) the outer diameter was large (~2.7 mm) and 2) available peak pressure rapidly decreased with increasing pulse number. In this study, we developed a new fiber delivery system to overcome these problems. As a laser absorbing material, we used a cap-type silicone rubber containing carbon black, into which the fiber output end can simply be inserted. The fiber end surface works to confine the laser-induced plasma. The outer diameter of the fiber system was reduced to ~1.4 mm. At an output laser fluence of 1.2 J/cm2, peak pressure of the first PMW pulse exceeded ~40 MPa. With successive 10 laser pulses, decreasing rate of the peak pressure was 22%, which was considerably lower than that with the previous fiber system (82%), enabling generation of at least successive 30 pulses of PMW with the same cap-type target. With this fiber system, we attempted transfer of plasmid DNA encoding EGFP (enhanced green fluorescence protein) to the rat skin as a test tissue in vivo, showing site-selective efficient gene expression.

  18. pH-responsive biocompatible fluorescent polymer nanoparticles based on phenylboronic acid for intracellular imaging and drug delivery

    Science.gov (United States)

    Li, Shengliang; Hu, Kelei; Cao, Weipeng; Sun, Yun; Sheng, Wang; Li, Feng; Wu, Yan; Liang, Xing-Jie

    2014-10-01

    To address current medical challenges, there is an urgent need to develop drug delivery systems with multiple functions, such as simultaneous stimuli-responsive drug release and real-time imaging. Biocompatible polymers have great potential for constructing smart multifunctional drug-delivery systems through grafting with other functional ligands. More importantly, novel biocompatible polymers with intrinsic fluorescence emission can work as theranostic nanomedicines for real-time imaging and drug delivery. Herein, we developed a highly fluorescent nanoparticle based on a phenylboronic acid-modified poly(lactic acid)-poly(ethyleneimine)(PLA-PEI) copolymer loaded with doxorubicin (Dox) for intracellular imaging and pH-responsive drug delivery. The nanoparticles exhibited superior fluorescence properties, such as fluorescence stability, no blinking and excitation-dependent fluorescence behavior. The Dox-loaded fluorescent nanoparticles showed pH-responsive drug release and were more effective in suppressing the proliferation of MCF-7 cells. In addition, the biocompatible fluorescent nanoparticles could be used as a tool for intracellular imaging and drug delivery, and the process of endosomal escape was traced by real-time imaging. These pH-responsive and biocompatible fluorescent polymer nanoparticles, based on phenylboronic acid, are promising tools for intracellular imaging and drug delivery.To address current medical challenges, there is an urgent need to develop drug delivery systems with multiple functions, such as simultaneous stimuli-responsive drug release and real-time imaging. Biocompatible polymers have great potential for constructing smart multifunctional drug-delivery systems through grafting with other functional ligands. More importantly, novel biocompatible polymers with intrinsic fluorescence emission can work as theranostic nanomedicines for real-time imaging and drug delivery. Herein, we developed a highly fluorescent nanoparticle based on a

  19. Clinical performance of fluorescence-based methods for detection of occlusal caries lesions in primary teeth

    Directory of Open Access Journals (Sweden)

    Laura Regina Antunes PONTES

    2017-11-01

    Full Text Available Abstract We aimed to investigate the performance of fluorescence-based methods (FBMs, compared to visual inspection after histological validation, in detecting and assessing the activity status of occlusal carious lesions in primary teeth. One examiner evaluated 50 primary molars close to exfoliation in 24 children. Teeth were assessed using quantitative light-induced fluorescence (QLF and pen-type laser fluorescence (LFpen. After exfoliation, histological validation was performed. Teeth were cut and sections were evaluated for lesion depth and activity status (after utilization of a pH indicator under a stereomicroscope. Parameters related to the performance of the methods in detecting caries lesions at two thresholds (initial and dentin lesions were calculated. Regarding the activity status, lesions were classified into sound+inactive or active, and the area under the ROC curve and the diagnostic odds ratio values of the methods were calculated and compared. Evaluation of red fluorescence using QLF presented higher sensitivity but lower specificity than visual inspection in detecting dentin caries lesions. However, QLF considering different parameters and LFpen had similar performance to that obtained with visual inspection. Regarding activity assessment, all FBMs and visual inspection also presented similar performance. In conclusion, FBMs did not prove advantageous for the detection and activity assessment of occlusal caries lesions in primary molars when compared to visual inspection.

  20. Fluorescent carbon nanoparticle-based lateral flow biosensor for ultrasensitive detection of DNA.

    Science.gov (United States)

    Takalkar, Sunitha; Baryeh, Kwaku; Liu, Guodong

    2017-12-15

    We report a fluorescent carbon nanoparticle (FCN)-based lateral flow biosensor for ultrasensitive detection of DNA. Fluorescent carbon nanoparticle with a diameter of around 15nm was used as a tag to label a detection DNA probe, which was complementary with the part of target DNA. A capture DNA probe was immobilized on the test zone of the lateral flow biosensor. Sandwich-type hybridization reactions among the FCN-labeled DNA probe, target DNA and capture DNA probe were performed on the lateral flow biosensor. In the presence of target DNA, FCNs were captured on the test zone of the biosensor and the fluorescent intensity of the captured FCNs was measured with a portable fluorescent reader. After systematic optimizations of experimental parameters (the components of running buffers, the concentration of detection DNA probe used in the preparation of FCN-DNA conjugates, the amount of FCN-DNA dispensed on the conjugate pad and the dispensing cycles of the capture DNA probes on the test-zone), the biosensor could detect a minimum concentration of 0.4 fM DNA. This study provides a rapid and low-cost approach for DNA detection with high sensitivity, showing great promise for clinical application and biomedical diagnosis. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Structural and dynamical aspects of skin studied by multiphoton excitation fluorescence microscopy-based methods.

    Science.gov (United States)

    Bloksgaard, Maria; Brewer, Jonathan; Bagatolli, Luis A

    2013-12-18

    This mini-review reports on applications of particular multiphoton excitation microscopy-based methodologies employed in our laboratory to study skin. These approaches allow in-depth optical sectioning of the tissue, providing spatially resolved information on specific fluorescence probes' parameters. Specifically, by applying these methods, spatially resolved maps of water dipolar relaxation (generalized polarization function using the 6-lauroyl-2-(N,N-dimethylamino)naphthale probe), activity of protons (fluorescence lifetime imaging using a proton sensitive fluorescence probe--2,7-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein) and diffusion coefficients of distinct fluorescence probes (raster imaging correlation spectroscopy) can be obtained from different regions of the tissue. Comparative studies of different tissue strata, but also between equivalent regions of normal and abnormal excised skin, including applications of fluctuation correlation spectroscopy on transdermal penetration of liposomes are presented and discussed. The data from the different studies reported reveal the intrinsic heterogeneity of skin and also prove these strategies to be powerful noninvasive tools to explore structural and dynamical aspects of the tissue. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Clinical performance of fluorescence-based methods for detection of occlusal caries lesions in primary teeth.

    Science.gov (United States)

    Pontes, Laura Regina Antunes; Novaes, Tatiane Fernandes; Moro, Bruna Lorena Pereira; Braga, Mariana Minatel; Mendes, Fausto Medeiros

    2017-11-06

    We aimed to investigate the performance of fluorescence-based methods (FBMs), compared to visual inspection after histological validation, in detecting and assessing the activity status of occlusal carious lesions in primary teeth. One examiner evaluated 50 primary molars close to exfoliation in 24 children. Teeth were assessed using quantitative light-induced fluorescence (QLF) and pen-type laser fluorescence (LFpen). After exfoliation, histological validation was performed. Teeth were cut and sections were evaluated for lesion depth and activity status (after utilization of a pH indicator) under a stereomicroscope. Parameters related to the performance of the methods in detecting caries lesions at two thresholds (initial and dentin lesions) were calculated. Regarding the activity status, lesions were classified into sound+inactive or active, and the area under the ROC curve and the diagnostic odds ratio values of the methods were calculated and compared. Evaluation of red fluorescence using QLF presented higher sensitivity but lower specificity than visual inspection in detecting dentin caries lesions. However, QLF considering different parameters and LFpen had similar performance to that obtained with visual inspection. Regarding activity assessment, all FBMs and visual inspection also presented similar performance. In conclusion, FBMs did not prove advantageous for the detection and activity assessment of occlusal caries lesions in primary molars when compared to visual inspection.

  3. A Graphene Oxide-Based Fluorescent Method for the Detection of Human Chorionic Gonadotropin.

    Science.gov (United States)

    Xia, Ning; Wang, Xin; Liu, Lin

    2016-10-13

    Human chorionic gonadotropin (hCG) has been regarded as a biomarker for the diagnosis of pregnancy and some cancers. Because the currently used methods (e.g., disposable Point of Care Testing (POCT) device) for hCG detection require the use of many less stable antibodies, simple and cost-effective methods for the sensitive and selective detection of hCG have always been desired. In this work, we have developed a graphene oxide (GO)-based fluorescent platform for the detection of hCG using a fluorescein isothiocyanate (FITC)-labeled hCG-specific binding peptide aptamer (denoted as FITC-PPLRINRHILTR) as the probe, which can be manufactured cheaply and consistently. Specifically, FITC-PPLRINRHILTR adsorbed onto the surface of GO via electrostatic interaction showed a poor fluorescence signal. The specific binding of hCG to FITC-PPLRINRHILTR resulted in the release of the peptide from the GO surface. As a result, an enhanced fluorescence signal was observed. The fluorescence intensity was directly proportional to the hCG concentration in the range of 0.05-20 IU/mL. The detection limit was found to be 20 mIU/mL. The amenability of the strategy to hCG analysis in biological fluids was demonstrated by assaying hCG in the urine samples.

  4. Determination of adenine based on the fluorescence recovery of the L-Tryptophan-Cu2+ complex

    Science.gov (United States)

    Duan, Ruilin; Li, Chunyan; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Yuan, Yusheng; Hu, Xiaoli

    2016-01-01

    A simple and sensitive method for determination of adenine was developed based on fluorescence quenching and recovery of L-Tryptophan (L-Trp). The fluorescence of L-Trp could efficiently quenched by copper ion compared with other common metal ions. Upon addition of adenine (Ade) in L-Trp-Cu(II) system, the fluorescence was reoccurred. Under the optimum conditions, the recovery fluorescence intensity was linearly correlated with the concentration of adenine in the range from 0.34 to 25.0 μmol L-1, with a correlation coefficient (R2) of 0.9994. The detection limit (3σ/k) was 0.046 μmol L-1, indicating that this method could applied to detect trace adenine. In this study, amino acids including L-Trp, D-Trp, L-Tyr, D-Tyr, L-Phe, D-Phe were investigated and only L-Trp could well chelated copper ion. Additionally, the mechanism of quench and recovery also were discussed and the method was successfully applied to detect the adenine in DNA with satisfactory results.

  5. Wireless fluorescence capsule for endoscopy using single photon-based detection

    Science.gov (United States)

    Al-Rawhani, Mohammed A.; Beeley, James; Cumming, David R. S.

    2015-12-01

    Fluorescence Imaging (FI) is a powerful technique in biological science and clinical medicine. Current FI devices that are used either for in-vivo or in-vitro studies are expensive, bulky and consume substantial power, confining the technique to laboratories and hospital examination rooms. Here we present a miniaturised wireless fluorescence endoscope capsule with low power consumption that will pave the way for future FI systems and applications. With enhanced sensitivity compared to existing technology we have demonstrated that the capsule can be successfully used to image tissue autofluorescence and targeted fluorescence via fluorophore labelling of tissues. The capsule incorporates a state-of-the-art complementary metal oxide semiconductor single photon avalanche detector imaging array, miniaturised optical isolation, wireless technology and low power design. When in use the capsule consumes only 30.9 mW, and deploys very low-level 468 nm illumination. The device has the potential to replace highly power-hungry intrusive optical fibre based endoscopes and to extend the range of clinical examination below the duodenum. To demonstrate the performance of our capsule, we imaged fluorescence phantoms incorporating principal tissue fluorophores (flavins) and absorbers (haemoglobin). We also demonstrated the utility of marker identification by imaging a 20 μM fluorescein isothiocyanate (FITC) labelling solution on mammalian tissue.

  6. Fluorescent sensors for selective detection of thiols: expanding the intramolecular displacement based mechanism to new chromophores.

    Science.gov (United States)

    Niu, Li-Ya; Zheng, Hai-Rong; Chen, Yu-Zhe; Wu, Li-Zhu; Tung, Chen-Ho; Yang, Qing-Zheng

    2014-03-21

    Biological thiols, including cysteine (Cys), homocystein (Hcy) and glutathione (GSH), play crucial roles in maintaining the appropriate redox status of biological systems. An abnormal level of biothiols is associated with different diseases, therefore, the discrimination between them is of great importance. Herein, we present two fluorescent sensors for selective detection of biothiols based on our recently reported intramolecular displacement mechanism. We expanded this mechanism to commercially available chromophores, 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) and heptamethine cyanine dye IR-780. The sensors operate by undergoing displacement of chloride by thiolate. The amino groups of Cys/Hcy further replace the thiolate to form amino-substituted products, which exhibit dramatically different photophysical properties compared to sulfur-substituted products from the reaction with GSH. NBD-Cl is highly selective towards Cys/Hcy and exhibits significant fluorescence enhancement. IR-780 showed a variation in its fluorescence ratio towards Cys over other thiols. Both of the sensors can be used for live-cell imaging of Cys. The wide applicability of the mechanism may provide a powerful tool for developing novel fluorescent sensors for selective detection of biothiols.

  7. A fluorescent carbon-dots-based mitochondria-targetable nanoprobe for peroxynitrite sensing in living cells.

    Science.gov (United States)

    Wu, Xiaoxue; Sun, Shan; Wang, Yuhui; Zhu, Jiali; Jiang, Kai; Leng, Yumin; Shu, Qinghai; Lin, Hengwei

    2017-04-15

    Mitochondria, the power generators in cell, are a primary organelle of oxygen consumption and a main source of reactive oxygen/nitrogen species (ROS/RNS). Peroxynitrite (ONOO - ), known as a kind of RNS, has been considered to be a significant factor in many cell-related biological processes, and there is great desire to develop fluorescent probes that can sensitively and selectively detect peroxynitrite in living cells. Herein, we developed a fluorescent carbon-dots (C-dots) based mitochondria-targetable nanoprobe with high sensitivity and selectivity for peroxynitrite sensing in living cells. The C-dots with its surface rich in amino groups was synthesized using o-phenylenediamine as carbon precursor, and it could be covalently conjugated with a mitochondria-targeting moiety, i.e. triphenylphosphonium (TPP). In the presence of peroxynitrite, the fluorescence of the constructed nanoprobe (C-dots-TPP) was efficiently quenched via a mechanism of photoinduced electron transfer (PET). The nanoprobe exhibited relatively high sensitivity (limit of detection: 13.5nM) and selectivity towards peroxynitrite in aqueous buffer. The performance of the nanoprobe for fluorescence imaging of peroxynitrite in mitochondria was investigated. The results demonstrated that the nanoprobe showed fine mitochondria-targeting ability and imaging contrast towards peroxynitrite in living cells. We anticipate that the proposed nanoprobe will provide a facile tool to explore the role played by peroxynitrite in cytobiology. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Quantitative generalized ratiometric fluorescence spectroscopy for turbid media based on probe encapsulated by biologically localized embedding

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Xiu-Fang; Chen, Zeng-Ping, E-mail: zpchen2002@hotmail.com; Cui, Yin-Yin; Hu, Yuan-Liang; Yu, Ru-Qin

    2016-05-19

    PEBBLE (probe encapsulated by biologically localized embedding) nanosensor encapsulating an intensity-based fluorescence indicator and an inert reference fluorescence dye inside the pores of stable matrix can be used as a generalized wavelength-ratiometric probe. However, the lack of an efficient quantitative model render the choices of inert reference dyes and intensity-based fluorescence indicators used in PEBBLEs based generalized wavelength-ratiometric probes rather limited. In this contribution, an extended quantitative fluorescence model was derived specifically for generalized wavelength-ratiometric probes based on PEBBLE technique (QFM{sub GRP}) with a view to simplify the design of PEBBLEs and hence further extend their application potentials. The effectiveness of QFM{sub GRP} has been tested on the quantitative determination of free Ca{sup 2+} in both simulated and real turbid media using a Ca{sup 2+} sensitive PEBBLE nanosensor encapsulating Rhod-2 and eosin B inside the micropores of stable polyacrylamide matrix. Experimental results demonstrated that QFM{sub GRP} could realize precise and accurate quantification of free Ca{sup 2+} in turbid samples, even though there is serious overlapping between the fluorescence excitation peaks of eosin B and Ca{sup 2+} bound Rhod-2. The average relative predictive error value of QFM{sub GRP} for the test simulated turbid samples was 5.9%, about 2–4 times lower than the corresponding values of partial least squares calibration model and the empirical ratiometric model based on the ratio of fluorescence intensities at the excitation peaks of Ca{sup 2+} bound Rhod-2 and eosin B. The recovery rates of QFM{sub GRP} for the real and spiked turbid samples varied from 93.1% to 101%, comparable to the corresponding results of atomic absorption spectrometry. - Highlights: • An advanced model was derived for generalized wavelength-ratiometric PEBBLEs. • The model can simplify the design of generalized wavelength

  9. A novel off–on fluorescence chemosensor for Ca2+ based on Rhodamine–Coumarin Schiff base derivative

    International Nuclear Information System (INIS)

    An, Jun-mei; Yang, Zheng-yin; Yan, Mi-hui; Li, Tian-rong

    2013-01-01

    A novel sensor, 7-Hydroxy-4-methylcoumarin-8-carbaldehyde-(rhodamine) hydrazone (l) has been synthesized and investigated as a fluorescence chemosensor for Ca 2+ in acetonitrile. The compound is found preferential binding to Ca 2+ in presence of large excess of other competitive ions with associated changes in its optical and fluorescence spectral behavior. Upon the addition of Ca 2+ , an overall emission change of 64-fold was observed. Photoinduced electron transfer (PET) process, coupled with the intramolecular charge transfer (ICT) process, are proposed to explain the observed spectral response. -- Highlights: ► A novel fluorescent sensor based on rhodamine and coumarin has been synthesized to detect Ca 2+ . ► Sensesor shows high sensitivity and selectivity to Ca 2+ with detection limit of 30 ppb.► The sensor can distinguish Ca 2+ and Mg 2+ effectively

  10. Anomalous Fluorescence Enhancement from Double Heterostructure 3D Colloidal Photonic Crystals–A Multifunctional Fluorescence-Based Sensor Platform

    Science.gov (United States)

    Eftekhari, Ehsan; Li, Xiang; Kim, Tak H.; Gan, Zongsong; Cole, Ivan S.; Zhao, Dongyuan; Kielpinski, Dave; Gu, Min; Li, Qin

    2015-01-01

    Augmenting fluorescence intensity is of vital importance to the development of chemical and biochemical sensing, imaging and miniature light sources. Here we report an unprecedented fluorescence enhancement with a novel architecture of multilayer three-dimensional colloidal photonic crystals self-assembled from polystyrene spheres. The new technique uses a double heterostructure, which comprises a top and a bottom layer with a periodicity overlapping the excitation wavelength (E) of the emitters, and a middle layer with a periodicity matching the fluorescence wavelength (F) and a thickness that supports constructive interference for the excitation wavelength. This E-F-E double heterostructure displays direction-dependent light trapping for both excitation and fluorescence, coupling the modes of photonic crystal with multiple-beam interference. The E-F-E double heterostructure renders an additional 5-fold enhancement to the extraordinary FL amplification of Rhodamine B in monolithic E CPhCs, and 4.3-fold acceleration of emission dynamics. Such a self-assembled double heterostructue CPhCs may find significant applications in illumination, laser, chemical/biochemical sensing, and solar energy harvesting. We further demonstrate the multi-functionality of the E-F-E double heterostructure CPhCs in Hg (II) sensing. PMID:26400503

  11. Anomalous Fluorescence Enhancement from Double Heterostructure 3D Colloidal Photonic Crystals--A Multifunctional Fluorescence-Based Sensor Platform.

    Science.gov (United States)

    Eftekhari, Ehsan; Li, Xiang; Kim, Tak H; Gan, Zongsong; Cole, Ivan S; Zhao, Dongyuan; Kielpinski, Dave; Gu, Min; Li, Qin

    2015-09-24

    Augmenting fluorescence intensity is of vital importance to the development of chemical and biochemical sensing, imaging and miniature light sources. Here we report an unprecedented fluorescence enhancement with a novel architecture of multilayer three-dimensional colloidal photonic crystals self-assembled from polystyrene spheres. The new technique uses a double heterostructure, which comprises a top and a bottom layer with a periodicity overlapping the excitation wavelength (E) of the emitters, and a middle layer with a periodicity matching the fluorescence wavelength (F) and a thickness that supports constructive interference for the excitation wavelength. This E-F-E double heterostructure displays direction-dependent light trapping for both excitation and fluorescence, coupling the modes of photonic crystal with multiple-beam interference. The E-F-E double heterostructure renders an additional 5-fold enhancement to the extraordinary FL amplification of Rhodamine B in monolithic E CPhCs, and 4.3-fold acceleration of emission dynamics. Such a self-assembled double heterostructure CPhCs may find significant applications in illumination, laser, chemical/biochemical sensing, and solar energy harvesting. We further demonstrate the multi-functionality of the E-F-E double heterostructure CPhCs in Hg (II) sensing.

  12. Molecular Basis for pH Sensitivity and Proton Transfer in Green Fluorescent Protein: Protonation and Conformational Substates from Electrostatic Calculations

    OpenAIRE

    Scharnagl, C.; Raupp-Kossmann, R.; Fischer, S.F.

    1999-01-01

    We performed a theoretical study to elucidate the coupling between protonation states and orientation of protein dipoles and buried water molecules in green fluorescent protein, a versatile biosensor for protein targeting. It is shown that the ionization equilibria of the wild-type green fluorescent protein-fluorophore and the internal proton-binding site E222 are mutually interdependent. Two acid-base transitions of the fluorophore occur in the presence of neutral (physiologic pH) and ionize...

  13. Multispectral Fluorescence Imaging During Robot-assisted Laparoscopic Sentinel Node Biopsy: A First Step Towards a Fluorescence-based Anatomic Roadmap.

    Science.gov (United States)

    van den Berg, Nynke S; Buckle, Tessa; KleinJan, Gijs H; van der Poel, Henk G; van Leeuwen, Fijs W B

    2017-07-01

    During (robot-assisted) sentinel node (SN) biopsy procedures, intraoperative fluorescence imaging can be used to enhance radioguided SN excision. For this combined pre- and intraoperative SN identification was realized using the hybrid SN tracer, indocyanine green- 99m Tc-nanocolloid. Combining this dedicated SN tracer with a lymphangiographic tracer such as fluorescein may further enhance the accuracy of SN biopsy. Clinical evaluation of a multispectral fluorescence guided surgery approach using the dedicated SN tracer ICG- 99m Tc-nanocolloid, the lymphangiographic tracer fluorescein, and a commercially available fluorescence laparoscope. Pilot study in ten patients with prostate cancer. Following ICG- 99m Tc-nanocolloid administration and preoperative lymphoscintigraphy and single-photon emission computed tomograpy imaging, the number and location of SNs were determined. Fluorescein was injected intraprostatically immediately after the patient was anesthetized. A multispectral fluorescence laparoscope was used intraoperatively to identify both fluorescent signatures. Multispectral fluorescence imaging during robot-assisted radical prostatectomy with extended pelvic lymph node dissection and SN biopsy. (1) Number and location of preoperatively identified SNs. (2) Number and location of SNs intraoperatively identified via ICG- 99m Tc-nanocolloid imaging. (3) Rate of intraoperative lymphatic duct identification via fluorescein imaging. (4) Tumor status of excised (sentinel) lymph node(s). (5) Postoperative complications and follow-up. Near-infrared fluorescence imaging of ICG- 99m Tc-nanocolloid visualized 85.3% of the SNs. In 8/10 patients, fluorescein imaging allowed bright and accurate identification of lymphatic ducts, although higher background staining and tracer washout were observed. The main limitation is the small patient population. Our findings indicate that a lymphangiographic tracer can provide additional information during SN biopsy based on ICG- 99m

  14. The use of nanocrystal quantum dot as fluorophore reporters in molecular beacon-based assays

    Science.gov (United States)

    Adegoke, Oluwasesan; Park, Enoch Y.

    2016-12-01

    The utilization of molecular beacon (MB) biosensor probes to detect nucleic acid targets has received enormous interest within the scientific community. This interest has been stimulated by the operational qualities of MB-based probes with respect to their unique sensitivity and specificity. The design of MB biosensors entails not only optimizing the sequence of the loop to hybridize with the nucleic acid target or optimization of the length of the stem to tune the sensitivity but also the selection of the appropriate fluorophore reporter to generate the signal transduction read-out upon hybridization of the probe with the target sequence. Traditional organic fluorescent dyes are mostly used for signal reporting in MB assays but their optical properties in comparison to semiconductor fluorescent quantum dot (Qdot) nanocrystals are at a disadvantage. This review highlights the progress made in exploiting Qdot as fluorophore reporters in MB-based assays with the aim of instigating further development in the field of Qdot-MB technology. The development reported to date indicates that unparalleled fluorescence signal reporting in MB-based assays can be achieved using well-constructed Qdot fluorophores.

  15. A rapid-screening approach to detect and quantify microplastics based on fluorescent tagging with Nile Red

    Science.gov (United States)

    Maes, Thomas; Jessop, Rebecca; Wellner, Nikolaus; Haupt, Karsten; Mayes, Andrew G.

    2017-03-01

    A new approach is presented for analysis of microplastics in environmental samples, based on selective fluorescent staining using Nile Red (NR), followed by density-based extraction and filtration. The dye adsorbs onto plastic surfaces and renders them fluorescent when irradiated with blue light. Fluorescence emission is detected using simple photography through an orange filter. Image-analysis allows fluorescent particles to be identified and counted. Magnified images can be recorded and tiled to cover the whole filter area, allowing particles down to a few micrometres to be detected. The solvatochromic nature of Nile Red also offers the possibility of plastic categorisation based on surface polarity characteristics of identified particles. This article details the development of this staining method and its initial cross-validation by comparison with infrared (IR) microscopy. Microplastics of different sizes could be detected and counted in marine sediment samples. The fluorescence staining identified the same particles as those found by scanning a filter area with IR-microscopy.

  16. A comparative study of oxygen transmission rates through polymer films based on fluorescence quenching

    DEFF Research Database (Denmark)

    Siró, Istvan; Plackett, David; Sommer-Larsen, Peter

    2010-01-01

    Information on oxygen permeability through polymer films is essential for some applications, especially in food packaging where the control of oxygen levels can be critical in avoiding food spoilage. A permeability testing device using fluorescence-based optical oxygen sensing was developed...... as a potential new instrument for measuring the oxygen permeability of packaging films. The fluorescence-based permeability tester was validated against two existing commercial oxygen permeability measuring devices, the Mocon Ox-Tran 2/20 and PBI-Dansensor OPT-5000. Oxygen transmission rates (OTR) of polylactide...... (PLA) and nanoclay-reinforced PLA films, as well as polyethylene/poly(ethylene terephthalate) (PE/PET) and polypropylene/poly(ethylene terephthalate) (PP/PET) laminated films were determined at 23°C and 50% relative humidity using each of these instruments. No significant differences were observed...

  17. Synthesis and cytotoxicity study of novel 3-(triazolyl)coumarins based fluorescent scaffolds.

    Science.gov (United States)

    Sinha, Sohini; Kumaran, Anuja Plavuvalapil; Mishra, Debasish; Paira, Priyankar

    2016-11-15

    Recently a choice of fluorescent bioimaging probes have been developed as medical diagnostic tools. Herein, we have introduced a series of coumarin-based target specific probes for cancer theranostic application which play a dual role in the field of both diagnosis and therapy. A fluorogenic version of 1,3-dipolar cycloaddition between azides and alkynes (DBCO) has been introduced to develop the triazolylcoumarin based fluorescent scaffolds. These scaffolds were screened for their anticancer activity against breast cancer (MCF7) and human epitheloid cervix carcinoma (HeLa) cell line. It was established that triazolylcoumarins (5c and 5d) are having electronegative substitution in the benzene ring displayed most effective anticancer profile in both the cell lines. Compounds 5a and 5d exhibited maximum quantum yield and strong cellular uptake in the MCF-7 cell line. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Fickian-Based Empirical Approach for Diffusivity Determination in Hollow Alginate-Based Microfibers Using 2D Fluorescence Microscopy and Comparison with Theoretical Predictions

    Directory of Open Access Journals (Sweden)

    Maryam Mobed-Miremadi

    2014-12-01

    Full Text Available Hollow alginate microfibers (od = 1.3 mm, id = 0.9 mm, th = 400 µm, L = 3.5 cm comprised of 2% (w/v medium molecular weight alginate cross-linked with 0.9 M CaCl2 were fabricated to model outward diffusion capture by 2D fluorescent microscopy. A two-fold comparison of diffusivity determination based on real-time diffusion of Fluorescein isothiocyanate molecular weight (FITC MW markers was conducted using a proposed Fickian-based approach in conjunction with a previously established numerical model developed based on spectrophotometric data. Computed empirical/numerical (Dempiricial/Dnumerical diffusivities characterized by small standard deviations for the 4-, 70- and 500-kDa markers expressed in m2/s are (1.06 × 10−9 ± 1.96 × 10−10/(2.03 × 10−11, (5.89 × 10−11 ± 2.83 × 10−12/(4.6 × 10−12 and (4.89 × 10−12 ± 3.94 × 10−13/(1.27 × 10−12, respectively, with the discrimination between the computation techniques narrowing down as a function of MW. The use of the numerical approach is recommended for fluorescence-based measurements as the standard computational method for effective diffusivity determination until capture rates (minimum 12 fps for the 4-kDa marker and the use of linear instead of polynomial interpolating functions to model temporal intensity gradients have been proven to minimize the extent of systematic errors associated with the proposed empirical method.

  19. Low molecular weight poly (2-dimethylamino ethylmethacrylate) polymers with controlled.positioned fluorescent labeling: Synthesis, characterization and in vitro interaction with human endothelial cells

    DEFF Research Database (Denmark)

    Flebus, Luca; Lombart, Francois; Sevrin, Chantal

    2015-01-01

    ) fluorescent labeled PDMAEMA of low molecular weight (Mw) (below 15 kDa), controlling the position and density of fluorescein.The second goal was to analyze the possible difference in uptake and subcellular distribution of this labeled FF polycation between human umbilical vein endothelial cells (HUVEC) and h...... to a minor cytotoxicity compared to the higher ones.As main conclusion this study highlights the similitude in cell trafficking of FF PDMAEMA and data previously reported for PDMAEMA/DNA complexes....

  20. Network-Based Models in Molecular Biology

    Science.gov (United States)

    Beyer, Andreas

    Biological systems are characterized by a large number of diverse interactions. Interaction maps have been used to abstract those interactions at all biological scales ranging from food webs at the ecosystem level down to protein interaction networks at the molecular scale.

  1. A Coumarin-Based Fluorescent Probe as a Central Nervous System Disease Biomarker

    OpenAIRE

    Yap, Ann-Chee; Mahamad, Ummi; Lim, Shen-Yang; Kim, Hae-Jo; Choo, Yeun-Mun

    2014-01-01

    Homocysteine and methylmalonic acid are important biomarkers for diseases associated with an impaired central nervous system (CNS). A new chemoassay utilizing coumarin-based fluorescent probe 1 to detect the levels of homocysteine is successfully implemented using Parkinson's disease (PD) patients' blood serum. In addition, a rapid identification of homocysteine and methylmalonic acid levels in blood serum of PD patients was also performed using the liquid chromatography-mass spectrometry (LC...

  2. Free-space fluorescence tomography with adaptive sampling based on anatomical information from microCT

    Science.gov (United States)

    Zhang, Xiaofeng; Badea, Cristian T.; Hood, Greg; Wetzel, Arthur W.; Stiles, Joel R.; Johnson, G. Allan

    2010-02-01

    Image reconstruction is one of the main challenges for fluorescence tomography. For in vivo experiments on small animals, in particular, the inhomogeneous optical properties and irregular surface of the animal make free-space image reconstruction challenging because of the difficulties in accurately modeling the forward problem and the finite dynamic range of the photodetector. These two factors are fundamentally limited by the currently available forward models and photonic technologies. Nonetheless, both limitations can be significantly eased using a signal processing approach. We have recently constructed a free-space panoramic fluorescence diffuse optical tomography system to take advantage of co-registered microCT data acquired from the same animal. In this article, we present a data processing strategy that adaptively selects the optical sampling points in the raw 2-D fluorescent CCD images. Specifically, the general sampling area and sampling density are initially specified to create a set of potential sampling points sufficient to cover the region of interest. Based on 3-D anatomical information from the microCT and the fluorescent CCD images, data points are excluded from the set when they are located in an area where either the forward model is known to be problematic (e.g., large wrinkles on the skin) or where the signal is unreliable (e.g., saturated or low signal-to-noise ratio). Parallel Monte Carlo software was implemented to compute the sensitivity function for image reconstruction. Animal experiments were conducted on a mouse cadaver with an artificial fluorescent inclusion. Compared to our previous results using a finite element method, the newly developed parallel Monte Carlo software and the adaptive sampling strategy produced favorable reconstruction results.

  3. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye.

    Science.gov (United States)

    Nath, Nidhi; Godat, Becky; Zimprich, Chad; Dwight, Stephen J; Corona, Cesear; McDougall, Mark; Urh, Marjeta

    2016-04-01

    Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer antibody therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of antibody drug conjugates (ADCs), efficient removal of surface receptors from cancer cells and changing the pharmacokinetics profile of the antibody drugs are some of key ways that internalization impacts the therapeutic efficacy of the antibodies. Over the years, several techniques have been used to study antibody internalization including radiolabels, fluorescent microscopy, flow cytometry and cellular toxicity assays. While these methods allow analysis of internalization, they have limitations including a multistep process and limited throughput and are generally endpoint assays. Here, we present a new homogeneous method that enables time and concentration dependent measurements of antibody internalization. The method uses a new hydrophilic and bright pH sensor dye (pHAb dye), which is not fluorescent at neutral pH but becomes highly fluorescent at acidic pH. For receptor mediated antibody internalization studies, antibodies against receptors are conjugated with the pHAb dye and incubated with the cells expressing the receptors. Upon binding to the receptor, the dyes conjugated to the antibody are not fluorescent because of the neutral pH of the media, but upon internalization and trafficking into endosomal and lysosomal vesicles the pH drops and dyes become fluorescent. The enabling attributes of the pHAb dyes are the hydrophilic nature to minimize antibody aggregation and bright fluorescence at acidic pH which allows development of simple plate based assays using a fluorescent reader. Using two different therapeutic antibodies--Trastuzumab (anti-HER2) and Cetuximab (anti-EGFR)--we show labeling with pHAb dye using amine and thiol chemistries and impact of chemistry and dye to antibody ration on internalization. We finally present two new approaches using the pHAb dye, which will be

  4. Detection of vegetation LUE based on chlorophyll fluorescence separation algorithm from Fraunhofer line

    Science.gov (United States)

    Liu, Liangyun; Zhang, Bing

    2009-09-01

    Photosynthetic efficiency is very important, and not yet generally assessable by remote sensing. Much research has proved the possibility of the separation of solar-induced chlorophyll fluorescence (ChlF) from the reflected hyperspectral data. As the 'probe' of plant photosynthesis, it is possible to detect photosynthetic light use efficiency (LUE) by the separated solar-induced ChlF. A diurnal experiment was carried out on winter wheat on Apr. 18, 2008, and the canopy radiance spectra and leaf LUE data were measured synchronously. The solar-induced chlorophyll fluorescence signals at 760nm and 688nm were separated from the reflected radiance spectral based on Fraunhofer lines in two oxygen absorption bands. The result showed that LUE was negatively correlated to the separated chlorophyll signals. The statistical models for LUE based on the solar-induced chlorophyll fluorescence values at 688 nm and 760 nm bands had correlation coefficients (R2) of 0.64 and 0.78, respectively. In addition, photochemical reflectance index (PRI) was also linked to LUE, and a statistical model for LUE based on PRI has a correlation coefficient (R2) of 0.66. The presented method provides a novel solution for monitoring LUE from remote sensing data.

  5. Pancreatic tumor detection using hypericin-based fluorescence spectroscopy and cytology

    Science.gov (United States)

    Lavu, Harish; Geary, Kevin; Fetterman, Harold R.; Saxton, Romaine E.

    2005-04-01

    Hypericin is a novel, highly fluorescent photosensitizer that exhibits selective tumor cell uptake properties and is particularly resistant to photobleaching. In this study, we have characterized hypericin uptake in human pancreatic tumor cells with relation to incubation time, cell number, and drug concentration. Ex vivo hypericin based fluorescence spectroscopy was performed to detect the presence of MIA PaCa-2 pancreatic tumor cells in the peritoneal cavity of BALB/c nude mice, as well as to quantify gross tumor burden. Hypericin based cytology of peritoneal lavage samples, using both one and two photon laser confocal microscopy, demonstrated more than a two-fold increase in fluorescence emission of pancreatic tumor cells as compared to control samples. In vitro treatment of pancreatic cancer cells with hypericin based photodynamic therapy showed tumor cell cytotoxicity in a drug dose, incident laser power, and time dependent manner. For these experiments, a continuous wavelength solid-state laser source (532 nm) was operated at power levels in the range of 100-400 mW. Potential applications of hypericin in tumor diagnosis, staging, and therapy will be presented.

  6. Irradiance Decay in Fluorescent and Light-emitting Diode-based Phototherapy Devices: A Pilot Study.

    OpenAIRE

    Olusanya, Bolajoko

    2016-01-01

    We set out to determine the rate of decline of irradiance for fluorescent tube (FT) and light-emitting diode (LED) phototherapy devices in resource-limited settings where routine irradiance monitoring is uncommon. Irradiance levels (μW/cm 2 /nm) were measured weekly using BiliBlanket ® II Meter on three FT-based and two LED-based phototherapy devices over a 19 week period. The two LED devices showed stable irradiance levels and did not require any lamp changes. The three FT-based devices show...

  7. Quantitative imaging of glutathione in live cells using a reversible reaction-based ratiometric fluorescent probe.

    Science.gov (United States)

    Jiang, Xiqian; Yu, Yong; Chen, Jianwei; Zhao, Mingkun; Chen, Hui; Song, Xianzhou; Matzuk, Alexander J; Carroll, Shaina L; Tan, Xiao; Sizovs, Antons; Cheng, Ninghui; Wang, Meng C; Wang, Jin

    2015-03-20

    Glutathione (GSH) plays an important role in maintaining redox homeostasis inside cells. Currently, there are no methods available to quantitatively assess the GSH concentration in live cells. Live cell fluorescence imaging revolutionized the field of cell biology and has become an indispensable tool in current biological studies. In order to minimize the disturbance to the biological system in live cell imaging, the probe concentration needs to be significantly lower than the analyte concentration. Because of this, any irreversible reaction-based GSH probe can only provide qualitative results within a short reaction time and will exhibit maximum response regardless of the GSH concentration if the reaction is completed. A reversible reaction-based probe with an appropriate equilibrium constant allows measurement of an analyte at much higher concentrations and, thus, is a prerequisite for GSH quantification inside cells. In this contribution, we report the first fluorescent probe-ThiolQuant Green (TQ Green)-for quantitative imaging of GSH in live cells. Due to the reversible nature of the reaction between the probe and GSH, we are able to quantify mM concentrations of GSH with TQ Green concentrations as low as 20 nM. Furthermore, the GSH concentrations measured using TQ Green in 3T3-L1, HeLa, HepG2, PANC-1, and PANC-28 cells are reproducible and well correlated with the values obtained from cell lysates. TQ Green imaging can also resolve the changes in GSH concentration in PANC-1 cells upon diethylmaleate (DEM) treatment. In addition, TQ Green can be conveniently applied in fluorescence activated cell sorting (FACS) to measure GSH level changes. Through this study, we not only demonstrate the importance of reaction reversibility in designing quantitative reaction-based fluorescent probes but also provide a practical tool to facilitate redox biology studies.

  8. Ratiometric Fluorescent Detection of Phosphate in Aqueous Solution Based on Near Infrared Fluorescent Silver Nanoclusters/Metal-Organic Shell Composite.

    Science.gov (United States)

    Dai, Cong; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2015-11-17

    Synthesis of near-infrared (NIR) fluorescent AgNCs with high quantum yield and stability is challenging but important for sensing and bioimaging application. Here, we report the fabrication of AgNCs/metal-organic shell composite via the deposition of metal-organic (zinc-nitrogen) coordination shell around AgNCs for ratiometric detection of phosphate. The composite exhibits NIR emission at 720 nm with 30 nm red-shift in comparison to bare AgNCs and a weak emission at 510 nm from the shell. The absolute quantum yield of NIR fluorescence of the composite is 15%, owing to FRET from the shell to the AgNCs core under the excitation at 430 nm. Besides, the composite is stable due to the protection of the shell. On the basis of the composite, a novel ratiometric fluorescence probe for the detection of phosphate in aqueous solution with good sensitivity and selectivity was developed. The limit of detection (3s) is 0.06 μM, and the relative standard deviation for 10 replicate detections of 10 μM phosphate was 0.6%. The recoveries of spiked phosphate in water, human urine, and serum samples ranged from 94.1% to 103.4%.

  9. [Department of the molecular bases of semiotics].

    Science.gov (United States)

    Ternovyĭ, K S

    1995-01-01

    Department of molecular basis of semiotics was organized in 1986. The main task of the department was to work out new approaches in estimation of the state of immune and blood system at the tissue, cell and molecular levels, using biochemical, biophysical and molecular biology techniques. There are several main directions of scientific investigations at the department. Most informational methods were collected in "immunological portrait" for differential diagnostic and complex investigation of the immune system of autoimmune patients. This group of techniques was used to study changes in the immune system of Kievites after the Chernobyl disaster. A decrease of complement and thymic serum activity was detected. Antibodies against nuclear components appeared in 20% of donors. And a higher of circulating immune complex of low molecular weight was observed. Low level of thymic serum activity in blood of autoimmune patients with rheumatoid arthritis, lupus erythematosus, diabetes, herpes and other depends on the appearance of zinc-independent timuline inhibitor less then 2000 D. Another kind of thymic hormone inhibitors was detected in thymectomized adult mice. Its effect disappears when zinc added in blood rather due to competition for lymphocyte surface receptors timuline and its inactive analogue than other mechanism. Therapeutic effect of UV irradiation of patients' blood was shown to be closely connected with the changes in thymic serum activity in respect to stabilization of thymic hormone/inhibitor ratio. The immunochemical techniques were used to detect and investigate tumor-associated chromatin antigens in human and animal tumor cells. Antigens not found in normal tissues were detected when using rabbit antibodies against chromatin of rat hepatocarcinoma and human colon and carcinoma.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Organic Material Based Fluorescent Sensor for Hg2+: a Brief Review on Recent Development.

    Science.gov (United States)

    Saleem, Muhammad; Rafiq, Muhammad; Hanif, Muhammad

    2017-01-01

    Due to the deleterious effects of mercury on human health and natural ecosystems, high reactivity, non-degradability, extreme volatility and relative water and tissue solubility, it would consider as one of the most toxic environmental pollutants among the transition metals. In the present investigation, we have tried to summarized the several organic material based fluorescent sensor including rhodamine, boron-dipyrromethene (BODIPYs), thiourea, crown-ether, coumarine, squaraines, pyrene, imidazole, triazole, anthracene, dansyl, naphthalenedimide/ naphthalene/ naphthalimide, naphthyridine, iridium (III) complexes, polymeric materials, cyclodextrin, phthalic anhydride, indole, calix [4]arene, chromenone, 1,8-naphthalimides, lysine, styrylindolium, phenothiazine, thiocarbonyl quinacridone, oxadiazole, triphenylamine-triazines, tetraphenylethene, peptidyl and semicarbazone for the trace mercury detection in the aqueous, aqueous-organic and cellular media. The present review provides a brief look over the previous development in the organic material based fluorescent sensor for mercuric ion detection. Furthermore, the ligand-metal binding stoichiometry, binding/association/dissociation constants and the detection limit by the receptors have been particularly highlighted which might be useful for the future design and development of more sensitive and robust fluorescent chemosensor/chemodosimeter for the mercuric ion detection. Graphical Abstract Dummy.

  11. Simple, fast, and low-cost camera-based water content measurement with colorimetric fluorescent indicator

    Science.gov (United States)

    Song, Seok-Jeong; Kim, Tae-Il; Kim, Youngmi; Nam, Hyoungsik

    2018-05-01

    Recently, a simple, sensitive, and low-cost fluorescent indicator has been proposed to determine water contents in organic solvents, drugs, and foodstuffs. The change of water content leads to the change of the indicator's fluorescence color under the ultra-violet (UV) light. Whereas the water content values could be estimated from the spectrum obtained by a bulky and expensive spectrometer in the previous research, this paper demonstrates a simple and low-cost camera-based water content measurement scheme with the same fluorescent water indicator. Water content is calculated over the range of 0-30% by quadratic polynomial regression models with color information extracted from the captured images of samples. Especially, several color spaces such as RGB, xyY, L∗a∗b∗, u‧v‧, HSV, and YCBCR have been investigated to establish the optimal color information features over both linear and nonlinear RGB data given by a camera before and after gamma correction. In the end, a 2nd order polynomial regression model along with HSV in a linear domain achieves the minimum mean square error of 1.06% for a 3-fold cross validation method. Additionally, the resultant water content estimation model is implemented and evaluated in an off-the-shelf Android-based smartphone.

  12. Secondary caries detection with a novel fluorescence-based camera system in vitro

    Science.gov (United States)

    Brede, Olivier; Wilde, Claudia; Krause, Felix; Frentzen, Matthias; Braun, Andreas

    2010-02-01

    The aim of the study was to assess the ability of a fluorescence based optical system to detect secondary caries. The optical detecting system (VistaProof) illuminates the tooth surfaces with blue light emitted by high power GaN-LEDs at 405 nm. Employing this almost monochromatic excitation, fluorescence is analyzed using a RGB camera chip and encoded in color graduations (blue - red - orange - yellow) by a software (DBSWIN), indicating the degree of caries destruction. 31 freshly extracted teeth with existing fillings and secondary caries were cleaned, excavated and refilled with the same kind of restorative material. 19 of them were refilled with amalgam, 12 were refilled with a composite resin. Each step was analyzed with the respective software and analyzed statistically. Differences were considered as statistically significant at p0.05). There was a significant difference between baseline measurements of the teeth primarily filled with composite resins and the refilled situation (p=0.014). There was also a significant difference between the non-excavated and the excavated group (Composite p=0.006, Amalgam p=0.018). The in vitro study showed, that the fluorescence based system allows detecting secondary caries next to composite resin fillings but not next to amalgam restorations. Cleaning of the teeth is not necessary, if there is no visible plaque. Further studies have to show, whether the system shows the same promising results in vivo.

  13. A Practical Solution for 77 K Fluorescence Measurements Based on LED Excitation and CCD Array Detector.

    Directory of Open Access Journals (Sweden)

    Jacob Lamb

    Full Text Available The fluorescence emission spectrum of photosynthetic microorganisms at liquid nitrogen temperature (77 K provides important insights into the organization of the photosynthetic machinery of bacteria and eukaryotes, which cannot be observed at room temperature. Conventionally, to obtain such spectra, a large and costly table-top fluorometer is required. Recently portable, reliable, and largely maintenance-free instruments have become available that can be utilized to accomplish a wide variety of spectroscopy-based measurements in photosynthesis research. In this report, we show how to build such an instrument in order to record 77K fluorescence spectra. This instrument consists of a low power monochromatic light-emitting diode (LED, and a portable CCD array based spectrometer. The optical components are coupled together using a fiber optic cable, and a custom made housing that also supports a dewar flask. We demonstrate that this instrument facilitates the reliable determination of chlorophyll fluorescence emission spectra for the cyanobacterium Synechocystis sp. PCC 6803, and the green alga Chlamydomonas reinhardtii.

  14. A Ratiometric Fluorescent Sensor for Cd2+ Based on Internal Charge Transfer

    Directory of Open Access Journals (Sweden)

    Dandan Cheng

    2017-11-01

    Full Text Available This work reports on a novel fluorescent sensor 1 for Cd2+ ion based on the fluorophore of tetramethyl substituted bis(difluoroboron-1,2-bis[(1H-pyrrol-2-ylmethylene]hydrazine (Me4BOPHY, which is modified with an electron donor moiety of N,N-bis(pyridin-2-ylmethylbenzenamine. Sensor 1 has absorption and emission in visible region, at 550 nm and 675 nm, respectively. The long wavelength spectral response makes it easier to fabricate the fluorescence detector. The sensor mechanism is based on the tunable internal charge transfer (ICT transition of molecule 1. Binding of Cd2+ ion quenches the ICT transition, but turns on the π − π transition of the fluorophore, thus enabling ratiometric fluorescence sensing. The limit of detection (LOD was projected down to 0.77 ppb, which is far below the safety value (3 ppb set for drinking water by World Health Organization. The sensor also demonstrates a high selectivity towards Cd2+ in comparison to other interferent metal ions.

  15. Fluorescence-based remote irradiation sensor in liquid-filled hollow-core photonic crystal fiber

    Energy Technology Data Exchange (ETDEWEB)

    Zeltner, R.; Russell, P. St.J. [Max Planck Institute for the Science of Light, Guenther-Scharowsky-Str. 1, 91058 Erlangen (Germany); Department of Physics, University of Erlangen-Nuremberg, Guenther-Scharowsky-Str. 1, 91058 Erlangen (Germany); Bykov, D. S.; Xie, S. [Max Planck Institute for the Science of Light, Guenther-Scharowsky-Str. 1, 91058 Erlangen (Germany); Euser, T. G. [Max Planck Institute for the Science of Light, Guenther-Scharowsky-Str. 1, 91058 Erlangen (Germany); Cavendish Laboratory, University of Cambridge, J. J. Thomson Avenue, Cambridge CB3 0HE (United Kingdom)

    2016-06-06

    We report an irradiation sensor based on a fluorescent “flying particle” that is optically trapped and propelled inside the core of a water-filled hollow-core photonic crystal fiber. When the moving particle passes through an irradiated region, its emitted fluorescence is captured by guided modes of the fiber core and so can be monitored using a filtered photodiode placed at the fiber end. The particle speed and position can be precisely monitored using in-fiber Doppler velocimetry, allowing the irradiation profile to be measured to a spatial resolution of ∼10 μm. The spectral response can be readily adjusted by appropriate choice of particle material. Using dye-doped polystyrene particles, we demonstrate detection of green (532 nm) and ultraviolet (340 nm) light.

  16. Protein A Detection Based on Quantum Dots-Antibody Bioprobe Using Fluorescence Coupled Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Lin Qiu

    2014-01-01

    Full Text Available In this report, fluorescence detection coupled capillary electrophoresis (CE-FL was used to detect Protein A. Antibody was first labeled with Cy5 and then mixed with quantum dots (QDs to form QDs-antibody bioprobe. Further, we observed fluorescence resonance energy transfer (FRET from QDs donor to Cy5 acceptor. The bioprobe was formed and brought QDs and Cy5 close enough to allow FRET to occur. After adding protein A, the FRET system was broken and caused the FRET signal to decrease. Thus, a new method for the determination of protein A was proposed based on the FRET signal changes. This study provides a new trail of thought for the detection of protein.

  17. A LabVIEW-Based Virtual Instrument System for Laser-Induced Fluorescence Spectroscopy.

    Science.gov (United States)

    Wu, Qijun; Wang, Lufei; Zu, Lily

    2011-01-01

    We report the design and operation of a Virtual Instrument (VI) system based on LabVIEW 2009 for laser-induced fluorescence experiments. This system achieves synchronous control of equipment and acquisition of real-time fluorescence data communicating with a single computer via GPIB, USB, RS232, and parallel ports. The reported VI system can also accomplish data display, saving, and analysis, and printing the results. The VI system performs sequences of operations automatically, and this system has been successfully applied to obtain the excitation and dispersion spectra of α-methylnaphthalene. The reported VI system opens up new possibilities for researchers and increases the efficiency and precision of experiments. The design and operation of the VI system are described in detail in this paper, and the advantages that this system can provide are highlighted.

  18. Fluorescent deep-blue and hybrid white emitting devices based on a naphthalene-benzofuran compound

    KAUST Repository

    Yang, Xiaohui

    2013-08-01

    We report the synthesis, photophysics and electrochemical properties of naphthalene-benzofuran compound 1 and its application in organic light emitting devices. Fluorescent deep-blue emitting devices employing 1 as the emitting dopant embedded in 4-4′-bis(9-carbazolyl)-2,2′-biphenyl (CBP) host show the peak external quantum efficiency of 4.5% and Commission Internationale d\\'Énclairage (CIE) coordinates of (0.15, 0.07). Hybrid white devices using fluorescent blue emitting layer with 1 and a phosphorescent orange emitting layer based on an iridium-complex show the peak external quantum efficiency above 10% and CIE coordinates of (0.31, 0.37). © 2013 Published by Elsevier B.V.

  19. In vitro and in vivo imaging of peroxynitrite by a ratiometric boronate-based fluorescent probe.

    Science.gov (United States)

    Palanisamy, Sathyadevi; Wu, Pei-Yu; Wu, Shou-Cheng; Chen, Yu-Jen; Tzou, Shey-Cherng; Wang, Chih-Hong; Chen, Chiao-Yun; Wang, Yun-Ming

    2017-05-15

    Peroxynitrite (ONOO - ) is an important species involved in many physiopathological processes. Progresses have been made in developing novel fluorescent probes to detect peroxynitrite with relatively high sensitivity and specificity. Herein, we report the synthesis, characterization and biological applications of a new boronate-based fluorescent probe, 4-MB. The studies showed that 4-MB exhibits a dual ratiometric and calorimetric response toward peroxynitrite due to ONOO - -triggered oxidative reaction. A possible mechanism of the oxidation reaction was proposed and the reaction product was isolated and characterized using different spectroscopic methods. We have thoroughly demonstrated the utility of 4-MB for intracellular peroxynitrite imaging. Further, we showed that 4-MB can be potentially employed to visualize exogenous and endogenous peroxynitrite in RAW264.7 macrophages, EAhy926 cells, zebrafish and in live tissues from a high-fat diet-induced obese mouse model. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Fluorescence-based lateral flow assays for rapid oral fluid roadside detection of cannabis use.

    Science.gov (United States)

    Plouffe, Brian D; Murthy, Shashi K

    2017-02-01

    With the recent worldwide changes in the legalization of marijuana, there is a significant need for rapid, roadside screening test for driving under the influence of drugs. A robust, sensitive, lateral flow assay has been developed to detect recent use via oral-fluid testing for Δ 9 -tetrahydrocannabinol (THC). This proof-of-concept assay uses a fluorescent-based immunoassay detection of polymeric beads, conjugated to antibodies against native THC. The fluorescent technique allows for significantly lower limits of detection and higher precision determination of recent marijuana use without the use of urine or blood sampling-thus allowing for roadside identification. Detection levels of 0.01 ng/mL were distinguished from background and the lower limit of quantification was determined to approach 1 ng/mL. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Molecular biology-based methods for quantification of bacteria in mixed culture: perspectives and limitations.

    Science.gov (United States)

    Nagarajan, Karthiga; Loh, Kai-Chee

    2014-08-01

    Species-specific enumeration of mixed community is invaluable as it facilitates a better understanding of the significance of the individual strains, their interactions, and the underlying mechanisms of community dynamics. Mixed microbial community has been characterized by microbiological, biochemical, or molecular biology-based methods. While microbiological and biochemical techniques do not provide adequate quantitative information of the members of the consortia and require additional techniques for a more comprehensive analysis, molecular biology-based methods analyze the microbial consortium based on specific DNA sequences and do not require isolation and culturing of bacteria for quantitative analysis. These methods outshine conventional culture-based techniques in terms of better sensitivity, reproducibility, and reliability. Quantitative molecular biology methods have been classified as PCR-based and probe hybridization methods. The PCR-based methods includes quantitative real-time PCR and terminal restriction fragment length polymorphism, while fluorescent in situ hybridization and DNA microarrays fall under probe hybridization methods. The workflow, the quantification methods, and their potential applications are discussed in this review by highlighting their advantages and possible limitations.

  2. Determinants of molecular marker based classification of rice (Oryza ...

    African Journals Online (AJOL)

    mr devi singh

    2015-01-07

    Jan 7, 2015 ... The present communication is aimed to find out determinants of molecular marker based classification of rice (Oryza sativa L) germplasm using the available data from an experiment conducted for development of molecular fingerprints of diverse varieties of Basmati and non Basmati rice adapted to.

  3. In Vivo Tumor Angiogenesis Imaging Using Peptide-Based Near-Infrared Fluorescent Probes.

    Science.gov (United States)

    Huang, Rui; Conti, Peter S; Chen, Kai

    2016-01-01

    Near-infrared fluorescence (NIRF) imaging is an emerging imaging technique for studying diseases at the molecular level. Optical imaging with a near-infrared emitting fluorophore for targeting tumor angiogenesis offers a noninvasive method for early tumor detection and efficient monitoring of tumor response to anti-angiogenesis therapy. CD13 receptor, a zinc-dependent membrane-bound ectopeptidase, plays important roles in regulating tumor angiogenesis and the growth of new blood vessels. In this chapter, we use CD13 receptor as an example to demonstrate how to construct CD13-specific NGR-containing peptides via bioorthogonal click chemistry for visualizing and quantifying the CD13 receptor expression in vivo by means of NIRF optical imaging.

  4. An “on-off-on” fluorescent nanoprobe for recognition of chromium(VI) and ascorbic acid based on phosphorus/nitrogen dual-doped carbon quantum dot

    Energy Technology Data Exchange (ETDEWEB)

    Gong, Xiaojuan, E-mail: gxj1124@sxu.edu.cn [Institute of Environmental Science, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Liu, Yang; Yang, Zhenhua; Shuang, Shaomin [Institute of Environmental Science, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Zhang, Zeyu [Faculty of Science, Beijing University of Chemical Technology, Beijing, 100029 (China); Dong, Chuan, E-mail: dc@sxu.edu.cn [Institute of Environmental Science, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China)

    2017-05-22

    Chromium (VI) [Cr(VI)] is a harsh environmental contaminates and has been proved to be highly toxic, carcinogenic and mutagenic. Therefore, developing an inexpensive, good selective and highly sensitive nanoprobe for the detection of Cr(VI) is in urgent demand. Recently, the highly fluorescent carbon quantum dots (CQDs) have been successfully utilized as efficient fluorescent nanoprobes for the detection of ions, pH and molecular substances. In this work, an “on-off” fluorescence phosphorus/nitrogen dual-doped CQDs (PNCQDs) probe was developed for the determination of Cr(VI) based on inner filter effect (IFE). The proposed PNCQDs nanoprobe shows its distinct merits of simplicity, convenience, fast implementation, good selectivity and high sensitivity towards Cr(VI), allowing its potential application in the determination of Cr(VI) in environment and biosystem. In addition, the chelation effect of the functional groups in reductant and Cr(VI), and the easy-conversion of Cr(VI) to reduced states (i.e. Cr(III) and Cr(0)) by reductants makes the minimization of IFE with a concomitant recovery of PNCQDs fluorescence possible. Hence, the PNCQDs/Cr(VI) hybrid was used as an “off-on” fluorescence probe for sensing ascorbic acid (AA), which is a model reductant. For the detection of Cr(VI), the linear range and the limit of detection achieved were 1.5–30 μmol/L and 23 nmol/L, respectively. For the detection of AA, the linear range and the limit of detection obtained were 5.0–200 μmol/L and 1.35 μmol/L, respectively. The as-constructed “on-off-on” PNCQDs fluorescent nanoprobe was successfully applied for detecting Cr(VI) and AA in biosystem. Furthermore, the as-constructed fluorescent sensing system was successfully applied to the analyses of AA in fresh fruits and in commercial fruit juices with satisfactory results. - Highlights: • Fast synthesis of phosphorus/nitrogen dual-doped CQDs (PNCQDs) by acid-base neutralization carbonization method.

  5. A fluorescent sensor based on binaphthol-quinoline Schiff base for ...

    Indian Academy of Sciences (India)

    ... complex was further used as a chemosensing ensemble for oxalate detection. The complex L₁-Zn²⁺ displays high selectivity to oxalate with significant fluorescence quenching through Zn²⁺ ion displacement approach. In addition, application of L₁ for imaging of Zn²⁺ and oxalate in living HeLa cells was also examined.

  6. Thin-film-based sensitivity enhancement for total internal reflection fluorescence live-cell imaging.

    Science.gov (United States)

    Kim, Kyujung; Cho, Eun-Jin; Huh, Yong-Min; Kim, Donghyun

    2007-11-01

    We investigated experimentally the evanescent field enhancement based on dielectric thin films in total internal reflection microscopy. The sample employed two layers of Al2O3 and SiO2 deposited on an SF10 glass substrate. Field intensity enhancement measured by fluorescent excitation of microbeads relative to that of a control sample without dielectric films was polarization dependent, determined as 4.2 and 2.4 for TE and TM polarizations, respectively, and was in good agreement with numerical results. The thin-film-based field enhancement was also applied to live-cell imaging of quantum dots, which confirmed the sensitivity enhancement qualitatively.

  7. Influence of water chemistry and dissolved organic matter (DOM) molecular size on copper and mercury binding determined by multiresponse fluorescence quenching.

    Science.gov (United States)

    Chen, W B; Smith, D S; Guéguen, C

    2013-07-01

    The effects of water chemistry (i.e. pH and Ca(2+) concentration) dissolved organic carbon (DOC) concentration) and DOM quality (i.e. composition and molecular weight) on metal complexation were successfully investigated by a combination of tangential flow filtration, excitation-emission matrix fluorescence, parallel factor analysis (PARAFAC), and fluorescence quenching on four freshwater samples and one extracted Suwannee River fulvic acid (SRFA). Two terrestrial and one microbial humic-like components were found in this study. Despite strong correlation between the Ryan-Weber model and the multiresponse model, the latter is more appropriate for the calculation of binding parameters in multiple-ligand DOM system. Decreasing pH from 6 to 4 significantly reduced logK-Cu(2+) from 5.22±0.24 to 4.60±0.30 at pH 6 and 4, respectively (pHigh molecular weight (>1kDa) DOM generally had higher logK and binding fluorophore abundance than bulk (unfractionated) and low molecular weight (<1kDa) DOM for both metals. This trend however was not always true for Hg(2+) where the binding parameters were quite variable. Overall the combined results provide evidence that binding parameters are not only affected by water chemistry, but also depend on DOM molecular weight. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Membranes and Fluorescence microscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2009-01-01

    Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence spectrosc......Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence...... spectroscopy approaches provide very valuable structurally and dynamically related information on membranes, they generally produce mean parameters from data collected on bulk solutions of many vesicles and lack direct information on the spatial organization at the level of single membranes, a quality that can...... be provided by microscopy-related techniques. In this chapter, I will attempt to summarize representative examples concerning how microscopy (which provides information on membrane lateral organization by direct visualization) and spectroscopy techniques (which provides information about molecular interaction...

  9. Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses

    Science.gov (United States)

    Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

    2016-01-01

    Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination. PMID:26877781

  10. Smartphone-Based Fluorescent Diagnostic System for Highly Pathogenic H5N1 Viruses.

    Science.gov (United States)

    Yeo, Seon-Ju; Choi, Kyunghan; Cuc, Bui Thi; Hong, Nguyen Ngoc; Bao, Duong Tuan; Ngoc, Nguyen Minh; Le, Mai Quynh; Hang, Nguyen Le Khanh; Thach, Nguyen Co; Mallik, Shyam Kumar; Kim, Hak Sung; Chong, Chom-Kyu; Choi, Hak Soo; Sung, Haan Woo; Yu, Kyoungsik; Park, Hyun

    2016-01-01

    Field diagnostic tools for avian influenza (AI) are indispensable for the prevention and controlled management of highly pathogenic AI-related diseases. More accurate, faster and networked on-site monitoring is demanded to detect such AI viruses with high sensitivity as well as to maintain up-to-date information about their geographical transmission. In this work, we assessed the clinical and field-level performance of a smartphone-based fluorescent diagnostic device with an efficient reflective light collection module using a coumarin-derived dendrimer-based fluorescent lateral flow immunoassay. By application of an optimized bioconjugate, a smartphone-based diagnostic device had a two-fold higher detectability as compared to that of the table-top fluorescence strip reader for three different AI subtypes (H5N3, H7N1, and H9N2). Additionally, in a clinical study of H5N1-confirmed patients, the smartphone-based diagnostic device showed a sensitivity of 96.55% (28/29) [95% confidence interval (CI): 82.24 to 99.91] and a specificity of 98.55% (68/69) (95% CI: 92.19 to 99.96). The measurement results from the distributed individual smartphones were wirelessly transmitted via short messaging service and collected by a centralized database system for further information processing and data mining. Smartphone-based diagnosis provided highly sensitive measurement results for H5N1 detection within 15 minutes. Because of its high sensitivity, portability and automatic reporting feature, the proposed device will enable agile identification of patients and efficient control of AI dissemination.

  11. Highly sensitive and selective fluorescent detection of cerebral lead(II) based on graphene quantum dot conjugates.

    Science.gov (United States)

    Qi, Yan-Xia; Zhang, Min; Fu, Qian-Qian; Liu, Ran; Shi, Guo-Yue

    2013-11-21

    A novel probe based on graphene quantum dot conjugates was developed for fluorescent detection of Pb(2+) (LOD: 9 pM) and furthermore for monitoring Pb(2+) in the striatum of rat, combined with microdialysis sampling in vivo.

  12. Fluorescence-based assay as a new screening tool for toxic chemicals

    Science.gov (United States)

    Moczko, Ewa; Mirkes, Evgeny M.; Cáceres, César; Gorban, Alexander N.; Piletsky, Sergey

    2016-09-01

    Our study involves development of fluorescent cell-based diagnostic assay as a new approach in high-throughput screening method. This highly sensitive optical assay operates similarly to e-noses and e-tongues which combine semi-specific sensors and multivariate data analysis for monitoring biochemical processes. The optical assay consists of a mixture of environmental-sensitive fluorescent dyes and human skin cells that generate fluorescence spectra patterns distinctive for particular physico-chemical and physiological conditions. Using chemometric techniques the optical signal is processed providing qualitative information about analytical characteristics of the samples. This integrated approach has been successfully applied (with sensitivity of 93% and specificity of 97%) in assessing whether particular chemical agents are irritating or not for human skin. It has several advantages compared with traditional biochemical or biological assays and can impact the new way of high-throughput screening and understanding cell activity. It also can provide reliable and reproducible method for assessing a risk of exposing people to different harmful substances, identification active compounds in toxicity screening and safety assessment of drugs, cosmetic or their specific ingredients.

  13. Breast cancer cells synchronous labeling and separation based on aptamer and fluorescence-magnetic silica nanoparticles

    Science.gov (United States)

    Wang, Qiu-Yue; Huang, Wei; Jiang, Xing-Lin; Kang, Yan-Jun

    2018-01-01

    In this work, an efficient method based on biotin-labeled aptamer and streptavidin-conjugated fluorescence-magnetic silica nanoprobes (FITC@Fe3O4@SiNPs-SA) has been established for human breast carcinoma MCF-7 cells synchronous labeling and separation. Carboxyl-modified fluorescence-magnetic silica nanoparticles (FITC@Fe3O4@SiNPs-COOH) were first synthesized using the Stöber method. Streptavidin (SA) was then conjugated to the surface of FITC@Fe3O4@SiNPs-COOH. The MCF-7 cell suspension was incubated with biotin-labeled MUC-1 aptamer. After centrifugation and washing, the cells were then treated with FITC@Fe3O4@SiNPs-SA. Afterwards, the mixtures were separated by a magnet. The cell-probe conjugates were then imaged using fluorescent microscopy. The results show that the MUC-1 aptamer could recognize and bind to the targeted cells with high affinity and specificity, indicating the prepared FITC@Fe3O4@SiNPs-SA with great photostability and superparamagnetism could be applied effectively in labeling and separation for MCF-7 cell in suspension synchronously. In addition, the feasibility of MCF-7 cells detection in peripheral blood was assessed. The results indicate that the method above is also applicable for cancer cells synchronous labeling and separation in complex biological system.

  14. A miniaturised image based fluorescence detection system for point-of-care-testing of cocaine abuse

    Science.gov (United States)

    Walczak, Rafał; Krüger, Jan; Moynihan, Shane

    2015-08-01

    In this paper, we describe a miniaturised image-based fluorescence detection system and demonstrate its viability as a highly sensitive tool for point-of-care-analysis of drugs of abuse in human sweat with a focus on monitor individuals for drugs of abuse. Investigations of miniaturised and low power optoelectronic configurations and methodologies for real-time image analysis were successfully carried out. The miniaturised fluorescence detection system was validated against a reference detection system under controlled laboratory conditions by analysing spiked sweat samples in dip stick and then strip with sample pad. As a result of the validation studies, a 1 ng mL-1 limit of detection of cocaine in sweat and full agreement of test results with the reference detection system can be reported. Results of the investigations open the way towards a detection system that integrates a hand-held fluorescence reader and a wearable skinpatch, and which can collect and in situ analyse sweat for the presence of cocaine at any point for up to tenths hours.

  15. Light emitting diode, photodiode-based fluorescence detection system for DNA analysis with microchip electrophoresis.

    Science.gov (United States)

    Hall, Gordon H; Glerum, D Moira; Backhouse, Christopher J

    2016-02-01

    Electrophoretic separation of fluorescently end-labeled DNA after a PCR serves as a gold standard in genetic diagnostics. Because of their size and cost, instruments for this type of analysis have had limited market uptake, particularly for point-of-care applications. This might be changed through a higher level of system integration and lower instrument costs that can be realized through the use of LEDs for excitation and photodiodes for detection--if they provide sufficient sensitivity. Here, we demonstrate an optimized microchip electrophoresis instrument using polymeric fluidic chips with fluorescence detection of end-labeled DNA with a LOD of 0.15 nM of Alexa Fluor 532. This represents orders of magnitude improvement over previously reported instruments of this type. We demonstrate the system with an electrophoretic separation of two PCR products and their respective primers. We believe that this is the first LED-induced fluorescence microchip electrophoresis system with photodiode-based detection that could be used for standard applications of PCR and electrophoresis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Optical tweezers and non-ratiometric fluorescent-dye-based studies of respiration in sperm mitochondria

    Science.gov (United States)

    Chen, Timothy; Shi, Linda Z.; Zhu, Qingyuan; Chandsawangbhuwana, Charlie; Berns, Michael W.

    2011-04-01

    The purpose of this study is to investigate how the mitochondrial membrane potential affects sperm motility using laser tweezers and a non-ratiometric fluorescent probe, DiOC6(3). A 1064 nm Nd:YVO4 continuous wave laser was used to trap motile sperm at a power of 450 mW in the trap spot. Using customized tracking software, the curvilinear velocity (VCL) and the escape force from the laser tweezers were measured. Human (Homo sapiens), dog (Canis lupis familiaris) and drill (Mandrillus leucophaeus) sperm were treated with DiOC6(3) to measure the membrane potential in the mitochondria-rich sperm midpieces. Sperm from all three species exhibited an increase in fluorescence when treated with the DiOC6(3). When a cyanide inhibitor (CCCP) of aerobic respiration was applied, sperm of all three species exhibited a reduction in fluorescence to pre-dye levels. With respect to VCL and escape force, the CCCP had no effect on dog or human sperm, suggesting a major reliance upon anaerobic respiration (glycolysis) for ATP in these two species. Based on the preliminary study on drill sperm, CCCP caused a drop in the VCL, suggesting potential reliance on both glycolysis and aerobic respiration for motility. The results demonstrate that optical trapping in combination with DiOC6(3) is an effective way to study sperm motility and energetics.

  17. An Intelligent Optical Dissolved Oxygen Measurement Method Based on a Fluorescent Quenching Mechanism

    Directory of Open Access Journals (Sweden)

    Fengmei Li

    2015-12-01

    Full Text Available Dissolved oxygen (DO is a key factor that influences the healthy growth of fishes in aquaculture. The DO content changes with the aquatic environment and should therefore be monitored online. However, traditional measurement methods, such as iodometry and other chemical analysis methods, are not suitable for online monitoring. The Clark method is not stable enough for extended periods of monitoring. To solve these problems, this paper proposes an intelligent DO measurement method based on the fluorescence quenching mechanism. The measurement system is composed of fluorescent quenching detection, signal conditioning, intelligent processing, and power supply modules. The optical probe adopts the fluorescent quenching mechanism to detect the DO content and solves the problem, whereas traditional chemical methods are easily influenced by the environment. The optical probe contains a thermistor and dual excitation sources to isolate visible parasitic light and execute a compensation strategy. The intelligent processing module adopts the IEEE 1451.2 standard and realizes intelligent compensation. Experimental results show that the optical measurement method is stable, accurate, and suitable for online DO monitoring in aquaculture applications.

  18. An Intelligent Optical Dissolved Oxygen Measurement Method Based on a Fluorescent Quenching Mechanism.

    Science.gov (United States)

    Li, Fengmei; Wei, Yaoguang; Chen, Yingyi; Li, Daoliang; Zhang, Xu

    2015-12-09

    Dissolved oxygen (DO) is a key factor that influences the healthy growth of fishes in aquaculture. The DO content changes with the aquatic environment and should therefore be monitored online. However, traditional measurement methods, such as iodometry and other chemical analysis methods, are not suitable for online monitoring. The Clark method is not stable enough for extended periods of monitoring. To solve these problems, this paper proposes an intelligent DO measurement method based on the fluorescence quenching mechanism. The measurement system is composed of fluorescent quenching detection, signal conditioning, intelligent processing, and power supply modules. The optical probe adopts the fluorescent quenching mechanism to detect the DO content and solves the problem, whereas traditional chemical methods are easily influenced by the environment. The optical probe contains a thermistor and dual excitation sources to isolate visible parasitic light and execute a compensation strategy. The intelligent processing module adopts the IEEE 1451.2 standard and realizes intelligent compensation. Experimental results show that the optical measurement method is stable, accurate, and suitable for online DO monitoring in aquaculture applications.

  19. Bioprobes Based on Aptamer and Silica Fluorescent Nanoparticles for Bacteria Salmonella typhimurium Detection

    Science.gov (United States)

    Wang, Qiu-Yue; Kang, Yan-Jun

    2016-03-01

    In this study, we have developed an efficient method based on single-stranded DNA (ssDNA) aptamers along with silica fluorescence nanoparticles for bacteria Salmonella typhimurium detection. Carboxyl-modified Tris(2,2'-bipyridyl)dichlororuthenium(II) hexahydrate (RuBPY)-doped silica nanoparticles (COOH-FSiNPs) were prepared using reverse microemulsion method, and the streptavidin was conjugated to the surface of the prepared COOH-FSiNPs. The bacteria S. typhimurium was incubated with a specific ssDNA biotin-labeled aptamer, and then the aptamer-bacteria conjugates were treated with the synthetic streptavidin-conjugated silica fluorescence nanoprobes (SA-FSiNPs). The results under fluorescence microscopy show that SA-FSiNPs can be applied effectively for the labeling of bacteria S. typhimurium with great photostable property. To further verify the specificity of SA-FSiNPs out of multiple bacterial conditions, variant concentrations of bacteria mixtures composed of bacteria S. typhimurium, Escherichia coli, and Bacillus subtilis were treated with SA-FSiNPs.

  20. Fiber optic-based fluorescence detection system for in vivo studies of exogenous chromophore pharmacokinetics

    Science.gov (United States)

    Doiron, Daniel R.; Dunn, J. B.; Mitchell, W. L.; Dalton, Brian K.; Garbo, Greta M.; Warner, Jon A.

    1995-05-01

    The detection and quantification of the concentration of exogenous chromophores in-vivo by their fluorescence is complicated by many physical and geometrical parameters. Measurement of such signals is advantageous in determining the pharmacokinetics of photosensitizers such as those used in photodynamic therapy (PDT) or to assist in the diagnosis of tissue histological state. To overcome these difficulties a ratio based fiber optic contact fluorometer has been developed. This fluorescence detection system (FDS) uses the ratio of the fluorescence emission peak of the exogenous chromophore to that of endogenous chromophores, i.e. autofluorescence, to correct for a variety of parameters affecting the magnitude of the measured signals. By doing so it also minimizes the range of baseline measurements prior to exogenous drug injection, for various tissue types. Design of the FDS and results of its testing in animals and patients using the second generation photosensitizer Tin ethyletiopurpurin (SnET2) are presented. These results support the feasibility and usefulness of the Ratio FDS system.

  1. Optical tweezers and non-ratiometric fluorescent-dye-based studies of respiration in sperm mitochondria

    International Nuclear Information System (INIS)

    Chen, Timothy; Shi, Linda Z; Zhu, Qingyuan; Chandsawangbhuwana, Charlie; Berns, Michael W

    2011-01-01

    The purpose of this study is to investigate how the mitochondrial membrane potential affects sperm motility using laser tweezers and a non-ratiometric fluorescent probe, DiOC 6 (3). A 1064 nm Nd:YVO4 continuous wave laser was used to trap motile sperm at a power of 450 mW in the trap spot. Using customized tracking software, the curvilinear velocity (VCL) and the escape force from the laser tweezers were measured. Human (Homo sapiens), dog (Canis lupis familiaris) and drill (Mandrillus leucophaeus) sperm were treated with DiOC 6 (3) to measure the membrane potential in the mitochondria-rich sperm midpieces. Sperm from all three species exhibited an increase in fluorescence when treated with the DiOC 6 (3). When a cyanide inhibitor (CCCP) of aerobic respiration was applied, sperm of all three species exhibited a reduction in fluorescence to pre-dye levels. With respect to VCL and escape force, the CCCP had no effect on dog or human sperm, suggesting a major reliance upon anaerobic respiration (glycolysis) for ATP in these two species. Based on the preliminary study on drill sperm, CCCP caused a drop in the VCL, suggesting potential reliance on both glycolysis and aerobic respiration for motility. The results demonstrate that optical trapping in combination with DiOC 6 (3) is an effective way to study sperm motility and energetics

  2. The effect of dimethylsulfoxide on absorption and fluorescence spectra of aqueous solutions of acridine orange base.

    Science.gov (United States)

    Markarian, Shiraz A; Shahinyan, Gohar A

    2015-12-05

    The photophysical properties of aqueous solutions of acridine orange base (AOB) in wide concentration range of dimethylsulfoxide (DMSO) were studied by using absorption and steady-state fluorescence spectroscopy techniques at room temperature. The absorption spectrum of acridine orange in water shows two bands at 468 and 490 nm which were attributed to the dimer ((AOBH)2(2+)) and monomer (AOBH(+)) species respectively. In DMSO solution for the same AOB concentration only the basic form was detected with the band at 428 nm. The addition of DMSO to AOB aqueous solution leads to the decrease of absorption band at 490 nm and the new absorption band increases at 428 nm due to deprotonated (basic) form of AO and the first isosbestic point occurs at 450 nm. The evolution of isosbestic point reveals that an other equilibrium, due to the self-association of DMSO molecules takes place. From the steady-state fluorescence spectra Stokes shifts were calculated for AOB in aqueous and DMSO solutions. The addition of DMSO into the aqueous solution induced the enhancement in the fluorescence intensity of the dye compared to those in water. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. A CTRW-based model of time-resolved fluorescence lifetime imaging in a turbid medium.

    Science.gov (United States)

    Chernomordik, Victor; Gandjbakhche, Amir H; Hassan, Moinuddin; Pajevic, Sinisa; Weiss, George H

    2010-12-01

    We develop an analytic model of time-resolved fluorescent imaging of photons migrating through a semi-infinite turbid medium bounded by an infinite plane in the presence of a single stationary point fluorophore embedded in the medium. In contrast to earlier models of fluorescent imaging in which photon motion is assumed to be some form of continuous diffusion process, the present analysis is based on a continuous-time random walk (CTRW) on a simple cubic lattice, the object being to estimate the position and lifetime of the fluorophore. Such information can provide information related to local variations in pH and temperature with potential medical significance. Aspects of the theory were tested using time-resolved measurements of the fluorescence from small inclusions inside tissue-like phantoms. The experimental results were found to be in good agreement with theoretical predictions provided that the fluorophore was not located too close to the planar boundary, a common problem in many diffusive systems.

  4. An efficient and biocompatible fluorescence resonance energy transfer system based on lanthanide-doped nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Di Weihua; Sakka, Yoshio [World Premier International Research (WPI) Center Initiative on Materials Nanoarchitronics (MANA), National Institute for Materials Science - NIMS, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Li Jie [Nanotechnology Innovation Center, NIMS, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan); Shirahata, Naoto, E-mail: weihdi@yahoo.com.cn, E-mail: SAKKA.Yoshio@nims.go.jp [Fine Particle Processing Group, Nano Ceramics Center, NIMS, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047 (Japan)

    2010-11-12

    This work demonstrates an efficient and bio-friendly fluorescence resonance energy transfer (FRET) system based on lanthanide-doped inorganic nanoparticles. A facile aqueous route was used to synthesize the CePO{sub 4}:Tb nanorods with homogeneous colloidal dispersion, which emits a bright green light with a high quantum yield ({approx}0.36) and a long fluorescence lifetime ({approx}3.50 ms) upon UV excitation. Upon treatment of CePO{sub 4}:Tb with aqueous Rhodamine B (RhB), an efficient FRET occurs from the Tb{sup 3+} to the RhB molecules, giving rise to well resolved and ratiometric emissions of donors and acceptors, respectively, with an energy transfer efficiency of up to 0.85. When incubated with HeLa cells at 37 deg. C, the CePO{sub 4}:Tb treated with RhB shows bright intracellular luminescence, indicating that it can be successfully internalized inside the cells and the FRET remains in the living cells. Moreover, the cytotoxic measurements demonstrate good biocompatibility and low cytotoxicity of our present FRET system. The advantages presented above including high quantum yield of donors, high energy transfer efficiency, ratiometric fluorescent emission and good biocompatibility, indicate the high potential of the CePO{sub 4}:Tb/RhB FRET system for monitoring biological events.

  5. Development of graphite carbon nitride based fluorescent immune sensor for detection of alpha fetoprotein

    Science.gov (United States)

    Li, Yike; Dong, Lingyu; Wang, Xiangfeng; Liu, Yuan; Liu, Hailing; Xie, Mengxia

    2018-05-01

    A novel fluorescent immunosensor for determination of alpha fetoprotein (AFP) in serum samples has been developed based on the nano graphite carbon nitride (g-C3N4) as fluorophore and immunomagnetic beads (MBs) as separation material. The bulk g-C3N4 was obtained by thermal polymerization of melamine, and then carboxylated and exfoliated to acquire the carboxylated nano g-C3N4 (c-n-g-C3N4), which has been characterized and the results showed that it had excellent fluorescent properties. The antibodies of AFP (Ab1, Ab2) were conjugated to the MBs and the c-n-g-C3N4, respectively. In assay of AFP detection, the magnetic part of the immunosensor, MBs-Ab1, would form the sandwich type complex with the signal part of the sensor, c-n-g-C3N4-Ab2. The developed immunosensor could simplify the process of separation due to the MBs. The results illustrated that proposed approach held a good linearity between the fluorescence intensity of the sensor and the AFP concentration ranging from 5-600 ng/mL with the limit of detection as low as 0.43 ng/mL, and its spiking recoveries ranged from 98.2% to 105.9% with RSD from 2.1% to 3.5%. The fabricated fluorescent immunosensor possesses the merits of good sensitivity, excellent selectivity, high biocompatibility and low cost, and the results provide a novel clue to develop immunosensor for determination of the biomarkers in complex matrices.

  6. Molecular recognition in poly(epsilon-caprolactone)-based thermoplastic elastomers

    NARCIS (Netherlands)

    Wisse, Eva; Spiering, A. J. H.; van Leeuwen, Ellen N. M.; Renken, Raymond A. E.; Dankers, Patricia Y. W.; Brouwer, Linda A.; van Luyn, Marja J. A.; Harmsen, Martin C.; Sommerdijk, Nico A. J. M.; Meijer, E. W.

    2006-01-01

    The molecular recognition properties of the hydrogen bonding segments in biodegradable thermoplastic elastomers were explored, aiming at the further functionalization of these potentially interesting biomaterials. A poly(epsilon-caprolactone)-based poly(urea) 2 was synthesized and characterized in

  7. Identification of cow and buffalo milk based on Beta carotene and vitamin-A concentration using fluorescence spectroscopy.

    Directory of Open Access Journals (Sweden)

    Rahat Ullah

    Full Text Available The current study presents the application of fluorescence spectroscopy for the identification of cow and buffalo milk based on β-carotene and vitamin-A which is of prime importance from the nutritional point of view. All samples were collected from healthy animals of different breeds at the time of lactation in the vicinity of Islamabad, Pakistan. Cow and buffalo milk shows differences at fluorescence emission appeared at band position 382 nm, 440 nm, 505 nm and 525 nm both in classical geometry (right angle setup as well as front face fluorescence setup. In front face fluorescence geometry, synchronous fluorescence emission shows clear differences at 410 nm and 440 nm between the milk samples of both these species. These fluorescence emissions correspond to fats, vitamin-A and β-carotene. Principal Component Analysis (PCA further highlighted these differences by showing clear separation between the two data sets on the basis of features obtained from their fluorescence emission spectra. These results indicate that classical geometry (fixed excitation wavelength as well as front face (synchronous fluorescence emission of cow and buffalo milk nutrients could be used as fingerprint from identification point of view. This same approach can effectively be used for the determination of adulterants in the milk and other dairy products.

  8. Identification of cow and buffalo milk based on Beta carotene and vitamin-A concentration using fluorescence spectroscopy.

    Science.gov (United States)

    Ullah, Rahat; Khan, Saranjam; Ali, Hina; Bilal, Muhammad; Saleem, Muhammad

    2017-01-01

    The current study presents the application of fluorescence spectroscopy for the identification of cow and buffalo milk based on β-carotene and vitamin-A which is of prime importance from the nutritional point of view. All samples were collected from healthy animals of different breeds at the time of lactation in the vicinity of Islamabad, Pakistan. Cow and buffalo milk shows differences at fluorescence emission appeared at band position 382 nm, 440 nm, 505 nm and 525 nm both in classical geometry (right angle) setup as well as front face fluorescence setup. In front face fluorescence geometry, synchronous fluorescence emission shows clear differences at 410 nm and 440 nm between the milk samples of both these species. These fluorescence emissions correspond to fats, vitamin-A and β-carotene. Principal Component Analysis (PCA) further highlighted these differences by showing clear separation between the two data sets on the basis of features obtained from their fluorescence emission spectra. These results indicate that classical geometry (fixed excitation wavelength) as well as front face (synchronous fluorescence emission) of cow and buffalo milk nutrients could be used as fingerprint from identification point of view. This same approach can effectively be used for the determination of adulterants in the milk and other dairy products.

  9. A-TEEMTM, a new molecular fingerprinting technique: simultaneous absorbance-transmission and fluorescence excitation-emission matrix method

    Science.gov (United States)

    Quatela, Alessia; Gilmore, Adam M.; Steege Gall, Karen E.; Sandros, Marinella; Csatorday, Karoly; Siemiarczuk, Alex; (Ben Yang, Boqian; Camenen, Loïc

    2018-04-01

    We investigate the new simultaneous absorbance-transmission and fluorescence excitation-emission matrix method for rapid and effective characterization of the varying components from a mixture. The absorbance-transmission and fluorescence excitation-emission matrix method uniquely facilitates correction of fluorescence inner-filter effects to yield quantitative fluorescence spectral information that is largely independent of component concentration. This is significant because it allows one to effectively monitor quantitative component changes using multivariate methods and to generate and evaluate spectral libraries. We present the use of this novel instrument in different fields: i.e. tracking changes in complex mixtures including natural water, wine as well as monitoring stability and aggregation of hormones for biotherapeutics.

  10. Dual fluorescent molecular substrates selectively report the activation, sustainability and reversibility of cellular PKB/Akt activity.

    Science.gov (United States)

    Shen, Duanwen; Bai, Mingfeng; Tang, Rui; Xu, Baogang; Ju, Xiaoming; Pestell, Richard G; Achilefu, Samuel

    2013-01-01

    Using a newly developed near-infrared (NIR) dye that fluoresces at two different wavelengths (dichromic fluorescence, DCF), we discovered a new fluorescent substrate for Akt, also known as protein kinase B, and a method to quantitatively report this enzyme's activity in real time. Upon insulin activation of cellular Akt, the enzyme multi-phosphorylated a single serine residue of a diserine DCF substrate in a time-dependent manner, culminating in monophospho- to triphospho-serine products. The NIR DCF probe was highly selective for the Akt1 isoform, which was demonstrated using Akt1 knockout cells derived from MMTV-ErbB2 transgenic mice. The DCF mechanism provides unparalleled potential to assess the stimulation, sustainability, and reversibility of Akt activation longitudinally. Importantly, NIR fluorescence provides a pathway to translate findings from cells to living organisms, a condition that could eventually facilitate the use of these probes in humans.

  11. Fluorescence-Based Comparative Binding Studies of the Supramolecular Host Properties of PAMAM Dendrimers Using Anilinonaphthalene Sulfonates: Unusual Host-Dependent Fluorescence Titration Behavior

    Directory of Open Access Journals (Sweden)

    Natasa Stojanovic

    2010-04-01

    Full Text Available This work describes the fluorescence enhancement of the anilinonaphthalene sulfonate probes 1,8-ANS, 2,6-ANS, and 2,6-TNS via complexation with PAMAM dendrimer hosts of Generation 4, 5 and 6. The use of this set of three very closely related probes allows for comparative binding studies, with specific pairs of probes differing only in shape (1,8-ANS and 2,6-ANS, or in the presence of a methyl substituent (2,6-TNS vs. 2,6-ANS. The fluorescence of all three probes was significantly enhanced upon binding with PAMAM dendrimers, however in all cases except one, a very unusual spike was consistently observed in the host fluorescence titration plots (fluorescence enhancement vs. host concentration at low dendrimer concentration. This unprecedented fluorescence titration curve shape makes fitting the data to a simple model such as 1:1 or 2:1 host: guest complexation very difficult; thus only qualitative comparisons of the relative binding of the three guests could be made based on host titrations. In the case of G4 and G5 dendrimers, the order of binding strength was qualitatively determined to be 1,8-ANS < 2,6-ANS indicating that the more streamlined 2,6-substituted probes are a better match for the dendrimer cavity shape than the bulkier 1,8-substituted probe. This order of binding strength was also indicated by double fluorometric titration experiments, involving both host and guest titrations. Further double fluorometric titration experiments on 2,6-ANS in G4 dendrimer revealed a host concentration-dependent change in the nature of the host: guest complexation, with multiple guests complexed per host molecule at very low host concentrations, but less than one guest per host at higher concentrations.

  12. Colorimetric assay for on-the-spot alcoholic strength sensing in spirit samples based on dual-responsive lanthanide coordination polymer particles with ratiometric fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Jingjing, E-mail: jjdeng@des.ecnu.edu.cn [School of Ecological and Environmental Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China); Shi, Guoyue [Department of Chemistry, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China); Zhou, Tianshu, E-mail: tszhou@des.ecnu.edu.cn [School of Ecological and Environmental Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China)

    2016-10-26

    This study demonstrates a new strategy for colorimetric detection of alcoholic strength (AS) in spirit samples based on dual-responsive lanthanide infinite coordination polymer (Ln-ICP) particles with ratiometric fluorescence. The ICP used in this study are composed of two components: one is the supramolecular Ln-ICP network formed by the coordination between the ligand 2,2’-thiodiacetic acid (TDA) and central metal ion Eu{sup 3+}; and the other is a fluorescent dye, i.e., coumarin 343 (C343), both as the cofactor ligand and as the sensitizer, doped into the Ln-ICP network through self-adaptive chemistry. Upon being excited at 300 nm, the red fluorescence of Ln-ICP network itself at 617 nm is highly enhanced due to the concomitant energy transfer from C343 to Eu{sup 3+}, while the fluorescence of C343 at 495 nm is supressed. In pure ethanol solvent, the as-formed C343@Eu-TDA is well dispersed and quite stable. However, the addition of water into ethanolic dispersion of C343@Eu-TDA destructs Eu-TDA network structure, resulting in the release of C343 from ICP network into the solvent. Consequently, the fluorescence of Eu-TDA turns off and the fluorescence of C343 turns on, leading to the fluorescent color change of the dispersion from red to blue, which constitutes a new mechanism for colorimetric sensing of AS in commercial spirit samples. With the method developed here, we could clearly distinguish the AS of different spirit samples within a wide linear range from 10% vol to 100% vol directly by “naked eye” with the help of UV-lamp (365 nm). This study not only offers a new method for on-the-spot visible detection of AS, but also provides a strategy for dual-responsive sensing mode by rational designing the optical properties of the Ln-ICP network and the guest, respectively. - Highlights: • Dual responsive lanthanide coordination polymer particles C343@Eu-TDA were synthesized. • The guest molecular coumarin 343 sensitized the luminescence of Eu

  13. Fluorescence perturbation techniques to study mobility and molecular dynamics of proteins in live cells: FRAP, photoactivation, photoconversion, and FLIP.

    Science.gov (United States)

    Bancaud, Aurélien; Huet, Sébastien; Rabut, Gwénaël; Ellenberg, Jan

    2010-12-01

    The technique of fluorescence recovery after photobleaching (FRAP) was introduced in the mid-1970s to study the diffusion of biomolecules in living cells. For several years, it was used mainly by a small number of biophysicists who had developed their own photobleaching systems. Since the mid-1990s, FRAP has gained increasing popularity because of the conjunction of two factors: First, photobleaching techniques are easily implemented on confocal laser-scanning microscopes (CLSMs), and so FRAP has become available to anyone who has access to such equipment. Second, the advent of green fluorescent protein (GFP) has allowed easy fluorescent tagging of proteins and their observation in living cells. Thanks both to the versatility of modern CLSMs, which allow control of laser intensity at any point of the image, and to the development of new fluorescent probes, additional photoperturbation techniques have emerged during the last few years. After the photoperturbation event, one observes and then analyzes how the fluorescence distribution relaxes toward the steady state. Because the photochemical perturbation of suitable fluorophores is essentially irreversible, changes of fluorescence intensity in the perturbed and unperturbed regions are due to the exchange of tagged molecules between those regions. This article first discusses the materials required for performing FRAP experiments on a CLSM and the software for data analysis. It then describes general considerations on how to perform FRAP experiments as well as the necessary controls. Finally, different possible ways to analyze the data are presented.

  14. Smartphone-Based Mobile Detection Platform for Molecular Diagnostics and Spatiotemporal Disease Mapping.

    Science.gov (United States)

    Song, Jinzhao; Pandian, Vikram; Mauk, Michael G; Bau, Haim H; Cherry, Sara; Tisi, Laurence C; Liu, Changchun

    2018-04-03

    Rapid and quantitative molecular diagnostics in the field, at home, and at remote clinics is essential for evidence-based disease management, control, and prevention. Conventional molecular diagnostics requires extensive sample preparation, relatively sophisticated instruments, and trained personnel, restricting its use to centralized laboratories. To overcome these limitations, we designed a simple, inexpensive, hand-held, smartphone-based mobile detection platform, dubbed "smart-connected cup" (SCC), for rapid, connected, and quantitative molecular diagnostics. Our platform combines bioluminescent assay in real-time and loop-mediated isothermal amplification (BART-LAMP) technology with smartphone-based detection, eliminating the need for an excitation source and optical filters that are essential in fluorescent-based detection. The incubation heating for the isothermal amplification is provided, electricity-free, with an exothermic chemical reaction, and incubation temperature is regulated with a phase change material. A custom Android App was developed for bioluminescent signal monitoring and analysis, target quantification, data sharing, and spatiotemporal mapping of disease. SCC's utility is demonstrated by quantitative detection of Zika virus (ZIKV) in urine and saliva and HIV in blood within 45 min. We demonstrate SCC's connectivity for disease spatiotemporal mapping with a custom-designed website. Such a smart- and connected-diagnostic system does not require any lab facilities and is suitable for use at home, in the field, in the clinic, and particularly in resource-limited settings in the context of Internet of Medical Things (IoMT).

  15. Controlling charge current through a DNA based molecular transistor

    Energy Technology Data Exchange (ETDEWEB)

    Behnia, S., E-mail: s.behnia@sci.uut.ac.ir; Fathizadeh, S.; Ziaei, J.

    2017-01-05

    Molecular electronics is complementary to silicon-based electronics and may induce electronic functions which are difficult to obtain with conventional technology. We have considered a DNA based molecular transistor and study its transport properties. The appropriate DNA sequence as a central chain in molecular transistor and the functional interval for applied voltages is obtained. I–V characteristic diagram shows the rectifier behavior as well as the negative differential resistance phenomenon of DNA transistor. We have observed the nearly periodic behavior in the current flowing through DNA. It is reported that there is a critical gate voltage for each applied bias which above it, the electrical current is always positive. - Highlights: • Modeling a DNA based molecular transistor and studying its transport properties. • Choosing the appropriate DNA sequence using the quantum chaos tools. • Choosing the functional interval for voltages via the inverse participation ratio tool. • Detecting the rectifier and negative differential resistance behavior of DNA.

  16. Quantification of Aerobic Ammonia-Oxidizing Bacteria in Soil using Activity-Based Fluorescent Labeling of Ammonia Monooxygenase

    Science.gov (United States)

    Farnan, J.; Bennett, K.; Hyman, M. R.

    2016-12-01

    Nitrification is a key step in the biological nitrogen cycle and has a large effect on the fate of nitrogen species in both wastewater treatment systems and agricultural soils. Aerobic ammonia-oxidizing bacteria (AOB) initiate nitrification by converting ammonia (NH3) to nitrite (NO2-) and are therefore pivotal to the process. AOB are ubiquitous in the environment but are difficult to quantify as they grow poorly on solid media. Other quantification methods like iquid most-probable number techniques are slow and error-prone, while modern molecular approaches involving polymerase chain reaction amplification are faster and more accurate but do not differentiate between active and inactive AOB. In this study, we explored using activity-based fluorescent mechanisms for rapidly quantifying metabolically active forms of AOB in soils. Initial experiments using Nitrosomonas europaea aimed to establish a relationship between NH3-dependent nitrite production and bacterial cell numbers. Active AMO was treated cells with 1,7-octadiyne (17OD) to inactivate the enzyme and a subsequent copper-dependent "click" reaction attached a fluor. The labeled protein was quantified by SDS-PAGE and IR scanning. In future experiments, AOB will be stimulated in soil microcosms by adding NH4Cl. AMO will again be inactivated by adding 17OD, and total bacteria will be separated from the soil samples using gradient centrifugation. After "click" conjugation with AlexaFluor 647 azide, the abundance of AMO will be determined with SDS-PAGE and IR analysis while metabolically active AOB will be measured via fluorescence-activating cell sorting.

  17. Effect of Moderate UVC Irradiation on Bovine Serum Albumin and Complex with Antimetabolite 5-Fluorouracil: Fluorescence Spectroscopic and Molecular Modelling Studies

    Directory of Open Access Journals (Sweden)

    Shanmugavel Chinnathambi

    2015-01-01

    Full Text Available The interaction of antimetabolite 5-fluorouracil (5FU with bovine serum albumin (BSA under UVC (253.7 nm irradiation was investigated in the present study using UV-Vis spectroscopy, steady state/time resolved fluorescence spectroscopic techniques. The stability of protein was found to be very strong when BSA gets bind to 5FU and moreover it is compared with the free BSA under UVC irradiation. From the fluorescence spectroscopic study, the stability of the complex was found to acquire 2-fold stronger than free protein. From the molecular modelling studies, we came to know the hydrogen bonds between BSA and antimetabolite 5FU are strong, up to 70.4 J/m2 under UVC irradiation.

  18. Bibliographic data base on atomic and molecular data

    International Nuclear Information System (INIS)

    Itikawa, Yukikazu.

    1983-03-01

    A comparative study is made on three bibliographic data bases: INSPEC, ORNL - AMPIC, GAPHYOR. An on - line retrieval is carried out for searching a number of specific atomic and molecular data. Characteristics of each data base are clarified and suggestions are given for use of those data bases. (author)

  19. Feasibility of the simultaneous determination of polycyclic aromatic hydrocarbons based on two-dimensional fluorescence correlation spectroscopy

    Science.gov (United States)

    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.

  20. Highly sensitive fluorescence resonance energy transfer (FRET)-based nanosensor for rapid detection of clenbuterol

    Science.gov (United States)

    Nghia Nguyen, Duc; Ngo, Trinh Tung; Liem Nguyen, Quang

    2012-09-01

    In this study we investigate the fabrication of a fluorescence resonance energy transfer (FRET)-based nanosensor for the detection of clenbuterol. The nanosensor consists of CdTe quantum dots coated by clenbuterol recognizable agent naphthol and diazotized clenbuterol. Changes in maximal photoluminescent intensities of the nanosensor were utilized to measure clenbuterol concentrations. The maximal photoluminescent intensities of the nanosensor were found to decrease with increasing clenbuterol concentrations, following a linear correlation. We have successfully fabricated a nanosensor for detection of clenbuterol with sensitivity up to 10 pg ml-1.

  1. Nanograting-based plasmon enhancement for total internal reflection fluorescence microscopy of live cells

    International Nuclear Information System (INIS)

    Kim, Kyujung; Cho, Eun-Jin; Suh, Jin-Suck; Huh, Yong-Min; Kim, Donghyun; Kim, Dong Jun

    2009-01-01

    We investigated evanescent field enhancement based on subwavelength nanogratings for improved sensitivity in total internal reflection microscopy of live cells. The field enhancement is associated with subwavelength-grating-coupled plasmon excitation. An optimum sample employed a silver grating on a silver film and an SF10 glass substrate. Field intensity was enhanced by approximately 90% when measured by fluorescent excitation of microbeads relative to that on a bare prism as a control, which is in good agreement with numerical results. The subwavelength-grating-mediated field enhancement was also applied to live cell imaging of quantum dots, which confirmed the sensitivity enhancement qualitatively.

  2. Multifunctional Material with Efficient Optoelectronic Integrated Molecular Switches Based on a Flexible Thin Film/Crystal.

    Science.gov (United States)

    Xu, Chang; Zhang, Wan-Ying; Ye, Qiong; Fu, Da-Wei

    2017-12-04

    Switchable materials, due to their potential applications in the fields of sensors, photonic devices, digital processing, etc., have been developed drastically. However, they still face great challenges in effectively inducing multiple molecular switching. Herein organic-inorganic hybrid compounds, an emerging class of hydrosoluble optoelectronic-active materials, welcome a new member with smart unique optical/electrical (fluorescence/dielectric) dual switches (switching ON/OFF), that is, [C 5 H 13 NBr][Cd 3 Br 7 ] (1) in the form of both a bulk crystal and an ultraflexible monodirectional thin film, which simultaneously exhibits fast dielectric/fluorescent dual switching triggered by an optical/thermal/electric signal with a high signal-to-noise ratio of 35 (the highest one in the known optical/dielectric dual molecular switches). Additionally, the exceptional stability/fatigue resistance as well as the fantastic extensibility/compactness of thin films (more than 10000 times folding over 90°), makes 1 an ideal candidate for single-molecule intelligent wearable devices and seamlessly integrated optoelectronic multiswitchable devices. This opens up a new route toward advanced light/electric high-performance switches/memories based on organic-inorganic hybrid compounds.

  3. Theory and development of fluorescence-based optochemical oxygen sensors: oxygen optodes.

    Science.gov (United States)

    Opitz, N; Lübbers, D W

    1987-01-01

    As the preceding considerations concerning the physical and technical features of oxygen optodes have demonstrated, fluorescence-based optochemical oxygen sensors possess certain advantages and peculiarities compared to conventionally applied electrochemical sensors such as polarographic oxygen electrodes. First, in contrast to oxygen electrodes, oxygen measurements with oxygen optodes do not suffer from distortions caused by the reference electrodes. In addition, because of the polarographic process, platinum electrodes continuously consume oxygen, which falsifies the results, especially when small sample volumes or long-term measurements, or both, are involved, whereas the sensor layer of oxygen optodes must only be equilibrated. Moreover, the surface of the platinum wire has to be catalytically clean in order to obtain a plateau of the polarogram and, consequently, to achieve a low rest current at zero PO2. Unfortunately, the demand for catalytically clean platinum surfaces turns out to be rather critical, since surface contamination occurs even with membranized electrodes, resulting in the well-known phenomenon of "electrode poisoning." The question of the specificity of oxygen electrodes also must be considered. In this context, CO2 and halothane may interfere with oxygen measurements, whereas fluorescence quenching is unaffected by CO2 and halothane affects the measurements only slightly, depending on the special indicator used. Furthermore, because of the flow dependence, oxygen measurements with the oxygen electrode show a distinct "stirring effect" caused by the turbulence in front of the electrode, which disturbs the diffusion field. Because of the completely different physical principle of fluorescence optical sensors, such influences are not observed with oxygen optodes. In addition, isolation and shielding of electrical circuits found in electrodes are not necessary for optodes. Furthermore, the sensitivity of oxygen optodes can be tuned to the desired

  4. Optimal separable bases and molecular collisions

    International Nuclear Information System (INIS)

    Poirier, L.W.

    1997-12-01

    A new methodology is proposed for the efficient determination of Green's functions and eigenstates for quantum systems of two or more dimensions. For a given Hamiltonian, the best possible separable approximation is obtained from the set of all Hilbert space operators. It is shown that this determination itself, as well as the solution of the resultant approximation, are problems of reduced dimensionality for most systems of physical interest. Moreover, the approximate eigenstates constitute the optimal separable basis, in the sense of self-consistent field theory. These distorted waves give rise to a Born series with optimized convergence properties. Analytical results are presented for an application of the method to the two-dimensional shifted harmonic oscillator system. The primary interest however, is quantum reactive scattering in molecular systems. For numerical calculations, the use of distorted waves corresponds to numerical preconditioning. The new methodology therefore gives rise to an optimized preconditioning scheme for the efficient calculation of reactive and inelastic scattering amplitudes, especially at intermediate energies. This scheme is particularly suited to discrete variable representations (DVR's) and iterative sparse matrix methods commonly employed in such calculations. State to state and cumulative reactive scattering results obtained via the optimized preconditioner are presented for the two-dimensional collinear H + H 2 → H 2 + H system. Computational time and memory requirements for this system are drastically reduced in comparison with other methods, and results are obtained for previously prohibitive energy regimes

  5. Application of cytoplasmic Ca2+ fluorescence imaging techniques to study the molecular mechanisms of exercise-induced fatigue eliminated by Chinese medicine ginseng extract

    Science.gov (United States)

    Liu, Yi; Zhao, Yanping; Zhang, Heming; Liu, Songhao

    2009-11-01

    The exercise-induced fatigue eliminated by Chinese medicine offers advantages including good efficiency and smaller side-effects, however, the exact mechanisms have not been classified. A lot of literatures indicated the cytosolic free Ca2+ concentrations of skeletal muscle cells increased significantly during exercise-induced fatigue. This study is aimed to establish a rat skeletal muscle cell model of exercise-induced fatigue. We applied cytoplasmic Ca2+ fluorescence imaging techniques to study the molecular mechanisms of exercise-induced fatigue eliminated by Chinese medicine ginseng extract. In our research, the muscle tissues from the newborn 3 days rats were taken out and digested into cells. The cells were randomly divided into the ginseng extract group and the control group. The cells from the two groups were cultured in the medium respectively added 2mg/ml ginseng extract and 2mg/ml D-hanks solution. After differentiating into myotubes, the two groups of cells treated with a fluorescent probe Fluo-3 AM were put on the confocal microscope and the fluorescence intensity of cells pre- and post- stimulation with dexamethasone were detected. It was found that cytoplasmic Ca2+ concentrations of the two groups of cells both increased post-stimulation, however, the increasing amplitude of fluorescence intensity of the ginseng extract group was significantly lower than that of the control group. In conclusion, stimulating the cells with dexamethasone is a kind of workable cell models of exercise-induced fatigue, and the molecular mechanisms of exercise-induced fatigue eliminated by ginseng extract may be connected to regulatating cytosolic free Ca2+ concentrations.

  6. Biomolecular imaging based on far-red fluorescent protein with a high two-photon excitation action cross section

    Science.gov (United States)

    Tsai, Tsung-Han; Lin, Cheng-Yung; Tsai, Huai-Jen; Chen, Szu-Yu; Tai, Shih-Peng; Lin, Kung-Hsuan; Sun, Chi-Kuang

    2006-04-01

    The two-photon excitation action cross section of Hc-Red fluorescent proteins (Hc-RFPs) is measured and found to be of the same order as that of enhanced green fluorescent proteins. With a 618 nm emission wavelength in the far-red region and with an excitation wavelength around 1200 nm, Hc-RPF-based two-photon fluorescence microscopy (2PFM) can offer deep penetration capability inside live samples and is ideal for in vivo gene expression study and biomolecular imaging in live objects. In vivo 2PFM of the developing heart deep inside a transgenic zebrafish embryo tagged by Hc-RFP is also successfully demonstrated.

  7. Steady State and Time-Resolved Fluorescence Dynamics of Triphenylamine Based Oligomers with Phenylene/Thiophene/Furan in Solvents

    International Nuclear Information System (INIS)

    Qi, Zeng; Ying-Liang, Liu; Kang, Meng; Xiang-Jie, Zhao; Shu-Feng, Wang; Qi-Huang, Gong

    2009-01-01

    We investigate the photo-physical properties of a series of triphenylamine-based oligomers by steady-state and picosecond transient fluorescence measurements in solvents. The oligomers are composed alternatively with triphenylamine and phenylene/thiophene/furan group, bridged by vinyl group (PNB/PNT/PNF). Their fluorescence spectra show bathochromic phenomenon with solvent polarity and viscosity increasing. The fluorescence decays are bi-exponential for PNB and PNT, and tri-exponential for PNF in THF and aniline. The strong viscosity dependence suggests conformational relaxation along the PNF chain after photo excitation. (condensed matter: electronicstructure, electrical, magnetic, and opticalproperties)

  8. Dipolar versus octupolar triphenylamine-based fluorescent organic nanoparticles as brilliant one- and two-photon emitters for (bio)imaging.

    Science.gov (United States)

    Parthasarathy, Venkatakrishnan; Fery-Forgues, Suzanne; Campioli, Elisa; Recher, Gaëlle; Terenziani, Francesca; Blanchard-Desce, Mireille

    2011-11-18

    Two related triphenylamine-based dipolar and octupolar fluorophores are used to prepare aqueous suspensions of fluorescent organic nanoparticles (FONs) via the reprecipitation method. The obtained spherical nanoparticles (30-40 nm in diameter) are fluorescent in aqueous solution (up to 15% fluorescence quantum yield) and exhibit extremely high one- and two-photon brightness, superior to those obtained for quantum dots. Despite the two chromophores showing similar fluorescence in solution, the fluorescence of FONs made from the octupolar derivative is significantly red-shifted compared to that generated by the dipolar FONs. In addition, the maximum two-photon absorption cross section of the FONs made from the octupolar derivative is 55% larger than that of the dipolar derivative FONs. The experimental observations provide evidence that the different molecular shape (rodlike versus three-branched) and charge distribution (dipolar versus octupolar) of the two chromophores strongly affect the packing inside the nanoparticles as well as their spectroscopic properties and colloidal stability in pure water. The use of these FONs as probes for biphotonic in-vivo imaging is investigated on Xenopus laevis tadpoles to test their utilization for angiography. When using FONs made from the octupolar dye, the formation of microagglomerates (2-5 μm scale) is observed in vivo, with subsequent lethal occlusion of the blood vessels. Conversely, the nanoparticles of the dipolar dye allow acute imaging of blood vessels thanks to their suitable size and brightness, while no toxic effect is observed. Such a goal cannot be achieved with the dissolved dye, which permeates the vessel walls. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Solid-state deep blue and UV fluorescent dyes based on para-bis(2-thienyl)phenylene

    Energy Technology Data Exchange (ETDEWEB)

    Krajčovič, Jozef; Kovalenko, Alexander, E-mail: kovalenko.alx@gmail.com; Heinrichová, Patricie; Vala, Martin; Weiter, Martin

    2015-11-15

    Despite the general rule of strong acceptor substituents having a tendency to quench fluorescence due to molecular stacking, it is shown how tetra-fluorination of the central phenylene unit of para-bis(2-thienyl)phenylene can augment the quantum yields of solid state fluorescent dyes. Another significant part of the present research was the study of the influence of the position of the solubilization alkyl chains position on the fluorescent properties of the abovementioned non- and tetra-fluorinated materials. Tenfold augmentation of quantum yields, depending on the position of the alkyl chains is reported. - Highlights: • Solid state luminescence was observed in para-bis(2-thienyl)phenylene molecules. • Quantum yields was augmented by polyfluorination of the central phenylene unit. • Tenfold augmentation of luminescence was observed by changing alkyls position. • Possibilities of steric hindrance and charge transfer were studied.

  10. Photoactivatable green fluorescent protein-based visualization and quantification of mitochondrial fusion and mitochondrial network complexity in living cells.

    Science.gov (United States)

    Karbowski, Mariusz; Cleland, Megan M; Roelofs, Brian A

    2014-01-01

    Technological improvements in microscopy and the development of mitochondria-specific imaging molecular tools have illuminated the dynamic rearrangements of these essential organelles. These rearrangements are mainly the result of two opposing processes: mitochondrial fusion and mitochondrial fission. Consistent with this, in addition to mitochondrial motility, these two processes are major factors determining the overall degree of continuity of the mitochondrial network, as well as the average size of mitochondria within the cell. In this chapter, we detail the use of advanced confocal microscopy and mitochondrial matrix-targeted photoactivatable green fluorescent protein (mito-PAGFP) for the investigation of mitochondrial dynamics. We focus on direct visualization and quantification of mitochondrial fusion and mitochondrial network complexity in living mammalian cells. These assays were instrumental in important recent discoveries within the field of mitochondrial biology, including the role of mitochondrial fusion in the activation of mitochondrial steps in apoptosis, participation of Bcl-2 family proteins in mitochondrial morphogenesis, and stress-induced mitochondrial hyperfusion. We present some basic directions that should be helpful in designing mito-PAGFP-based experiments. Furthermore, since analyses of mitochondrial fusion using mito-PAGFP-based assays rely on time-lapse imaging, critical parameters of time-lapse microscopy and cell preparation are also discussed.

  11. Fluorescence-based retention assays reveals sustained release of vascular endothelial growth factor from bone grafts.

    Science.gov (United States)

    Kang, Wonmo; Yun, Ye-Rang; Lee, Dong-Sung; Kim, Tae-Hyun; Kim, Joong-Hyun; Kim, Hae-Won; Jang, Jun-Hyeog

    2016-01-01

    The sustained release of growth factors following their implantation in vivo is essential for successful outcomes in bone tissue engineering. In this study, we evaluated the release kinetics and delivery efficacies of vascular endothelial growth factor (VEGF), a potent angiogenic growth factor, incorporated into calcium phosphate bone grafts (BGs). We evaluated the release profile of VEGF from BGs using a novel fluorescence-based retention assay, which revealed that VEGF loaded on BGs can be released in a sustained manner without an initial burst (near zero-order cumulative release) with a controlled release rate of 13.6% per week for up to 7 weeks. In contrast, an ELISA-based release assay showed VEGF to have an early burst-release profile for the first week. However, the biological activity of VEGF released from the BGs was preserved over the 7-week release period, which is consistent with the sustained-release profile observed in the fluorescence-based retention assay. Furthermore, the in vivo bone-forming action of the VEGF-loaded BGs was well demonstrated in a rat subcutaneous model. Taken together, the sustained release of VEGF loaded onto BGs was effective in stimulating proliferation, angiogenesis and osteogenesis, suggesting the ultimate value of VEGF-engineered BGs for bone tissue engineering. © 2015 Wiley Periodicals, Inc.

  12. Basic dose response of fluorescent screen-based portal imaging device

    International Nuclear Information System (INIS)

    Yeo, In Hwan; Yonannes, Yonas; Zhu, Yunping

    1999-01-01

    The purpose of this study is to investigate fundamental aspects of the dose response of fluorescent screen-based electronic portal imaging devices (EPIDs). We acquired scanned signal across portal planes as we varied the radiation that entered the EPID by changing the thickness and anatomy of the phantom as well as the air gap between the phantom and the EPID. In addition, we simulated the relative contribution of the scintillation light signal in the EPID system. We have shown that the dose profile across portal planes is a function of the air gap and phantom thickness. We have also found that depending on the density change within the phantom geometry, errors associated with dose response based on the EPID scan can be as high as 7%. We also found that scintillation light scattering within the EPID system is an important source of error. This study revealed and demonstrated fundamental characteristics of dose response of EPID, as relative to that of ion chambers. This study showed that EPID based on fluorescent screen cannot be an accurate dosimetry system

  13. An evaluation of commercial fluorescent bead-based luminex cytokine assays.

    Directory of Open Access Journals (Sweden)

    Joel Fleury Djoba Siawaya

    Full Text Available The recent introduction of fluorescent bead-based technology, allowing the measurement of multiples analytes in a single 25-50 microl sample has revolutionized the study of cytokine responses. However, such multiplex approaches may compromise the ability of these assays to accurately measure actual cytokine levels. This study evaluates the performance of three commercially available multiplex cytokine fluorescent bead-based immunoassays (Bio-Rad's Cytokine 17-plex kit; LINCO Inc's 29-plex kit; and RnD System's Fluorokine-Multi Analyte Profiling (MAP base kit A and B. The LINCO Inc kit was found to be the most sensitive assay for measuring concentrations of multiple recombinant cytokines in samples that had been spiked with serial dilutions of the standard provided by the manufacturer, followed respectively by the RnD Fluorokine-(MAP and Bio-Rad 17-plex kits. A positive correlation was found in the levels of IFN-gamma measured in antigen stimulated whole blood culture supernatants by the LINCO Inc 29-plex, RnD Fluorokine-(MAP and RnD system IFN-gamma Quantikine ELISA kits across a panel of controls and stimulated samples. Researchers should take the limitation of such multiplexed assays into account when planning experiments and the most appropriate use for these tests may currently be as screening tools for the selection of promising markers for analysis by more sensitive techniques.

  14. On-bead fluorescent DNA nanoprobes to analyze base excision repair activities

    Energy Technology Data Exchange (ETDEWEB)

    Gines, Guillaume; Saint-Pierre, Christine; Gasparutto, Didier, E-mail: didier.gasparutto@cea.fr

    2014-02-17

    Graphical abstract: -- Highlights: •On magnetic beads fluorescent enzymatic assays. •Simple, easy, non-radioactive and electrophoresis-free functional assay. •Lesion-containing hairpin DNA probes are selective for repair enzymes. •The biosensing platform allows the measurement of DNA repair activities from purified enzymes or within cell free extracts. -- Abstract: DNA integrity is constantly threatened by endogenous and exogenous agents that can modify its physical and chemical structure. Changes in DNA sequence can cause mutations sparked by some genetic diseases or cancers. Organisms have developed efficient defense mechanisms able to specifically repair each kind of lesion (alkylation, oxidation, single or double strand break, mismatch, etc). Here we report the adjustment of an original assay to detect enzymes’ activity of base excision repair (BER), that supports a set of lesions including abasic sites, alkylation, oxidation or deamination products of bases. The biosensor is characterized by a set of fluorescent hairpin-shaped nucleic acid probes supported on magnetic beads, each containing a selective lesion targeting a specific BER enzyme. We have studied the DNA glycosylase alkyl-adenine glycosylase (AAG) and the human AP-endonuclease (APE1) by incorporating within the DNA probe a hypoxanthine lesion or an abasic site analog (tetrahydrofuran), respectively. Enzymatic repair activity induces the formation of a nick in the damaged strand, leading to probe's break, that is detected in the supernatant by fluorescence. The functional assay allows the measurement of DNA repair activities from purified enzymes or in cell-free extracts in a fast, specific, quantitative and sensitive way, using only 1 pmol of probe for a test. We recorded a detection limit of 1 μg mL{sup −1} and 50 μg mL{sup −1} of HeLa nuclear extracts for APE1 and AAG enzymes, respectively. Finally, the on-bead assay should be useful to screen inhibitors of DNA repair

  15. On-bead fluorescent DNA nanoprobes to analyze base excision repair activities

    International Nuclear Information System (INIS)

    Gines, Guillaume; Saint-Pierre, Christine; Gasparutto, Didier

    2014-01-01

    Graphical abstract: -- Highlights: •On magnetic beads fluorescent enzymatic assays. •Simple, easy, non-radioactive and electrophoresis-free functional assay. •Lesion-containing hairpin DNA probes are selective for repair enzymes. •The biosensing platform allows the measurement of DNA repair activities from purified enzymes or within cell free extracts. -- Abstract: DNA integrity is constantly threatened by endogenous and exogenous agents that can modify its physical and chemical structure. Changes in DNA sequence can cause mutations sparked by some genetic diseases or cancers. Organisms have developed efficient defense mechanisms able to specifically repair each kind of lesion (alkylation, oxidation, single or double strand break, mismatch, etc). Here we report the adjustment of an original assay to detect enzymes’ activity of base excision repair (BER), that supports a set of lesions including abasic sites, alkylation, oxidation or deamination products of bases. The biosensor is characterized by a set of fluorescent hairpin-shaped nucleic acid probes supported on magnetic beads, each containing a selective lesion targeting a specific BER enzyme. We have studied the DNA glycosylase alkyl-adenine glycosylase (AAG) and the human AP-endonuclease (APE1) by incorporating within the DNA probe a hypoxanthine lesion or an abasic site analog (tetrahydrofuran), respectively. Enzymatic repair activity induces the formation of a nick in the damaged strand, leading to probe's break, that is detected in the supernatant by fluorescence. The functional assay allows the measurement of DNA repair activities from purified enzymes or in cell-free extracts in a fast, specific, quantitative and sensitive way, using only 1 pmol of probe for a test. We recorded a detection limit of 1 μg mL −1 and 50 μg mL −1 of HeLa nuclear extracts for APE1 and AAG enzymes, respectively. Finally, the on-bead assay should be useful to screen inhibitors of DNA repair activities

  16. Synthesis and Binding Ability of Molecular Probes Based on a Phenanthroline Derivative: Theory and Experiment

    Directory of Open Access Journals (Sweden)

    Xuefang Shang

    2013-12-01

    Full Text Available A fluorescent and colorimetric molecular probe containing phenol groups has been designed and synthesized. The anion binding ability was evaluated for biolgically important anions (F−, Cl−, Br−, I−, AcO− and H2PO4− by theoretical investigation, UV-Vis and fluorescence spectroscopy and 1H-NMR titration experiments. Results indicated the probe showed strong binding ability for H2PO4− without the interference of other anions tested and the interaction process was accompanied by color changes. Theoretical investigation analysis revealed that intramolecular hydrogen bonds existed in the structure of the probe and the roles of molecular frontier orbitals in molecular interplay were determined.

  17. Light scattering sensing detection of pathogens based on the molecular recognition of immunoglobulin with cell wall-associated protein A

    International Nuclear Information System (INIS)

    Liu Zhongde; Chen Shaofen; Cheng Zhihuang; Zhen Shujun; Liao Qiegen

    2007-01-01

    In this contribution, we report a rapid optical detection method of pathogens using Staphylococcus aureus (S. aureus) as the model analyte based on the molecular recognition of immunoglobulin with cell wall-associated Protein A (SpA). It was found that the molecular recognition of human immunoglobulin (IgG) with protein A on the cell wall of S. aureus on glass slide sensing area could result in strong surface enhanced light scattering (SELS) signals, and the SELS intensity (ΔI) increases proportionally with the concentration of S. aureus over the range of 2.5 x 10 5 -1.0 x 10 8 CFU mL -1 with right angle light scattering (RALS) signals detection mode. In order to identify the solid support based molecular recognition between IgG with SpA, we also employed water-soluble CdS quantum dots (CdS-QDs) as a fluorescent marker for IgG by immobilizing the IgG onto the surfaces of CdS-QDs through covalent binding in order to generate recognition probes for SpA on the cell wall of S. aureus. Consequently, the fluorescent method also showed that the detection for pathogens with solid supports is reliable based on the molecular recognition of IgG with SpA

  18. Green fluorescence protein-based content-mixing assay of SNARE-driven membrane fusion.

    Science.gov (United States)

    Heo, Paul; Kong, Byoungjae; Jung, Young-Hun; Park, Joon-Bum; Shin, Jonghyeok; Park, Myungseo; Kweon, Dae-Hyuk

    2017-06-17

    Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins mediate intracellular membrane fusion by forming a ternary SNARE complex. A minimalist approach utilizing proteoliposomes with reconstituted SNARE proteins yielded a wealth of information pinpointing the molecular mechanism of SNARE-mediated fusion and its regulation by accessory proteins. Two important attributes of a membrane fusion are lipid-mixing and the formation of an aqueous passage between apposing membranes. These two attributes are typically observed by using various fluorescent dyes. Currently available in vitro assay systems for observing fusion pore opening have several weaknesses such as cargo-bleeding, incomplete removal of unencapsulated dyes, and inadequate information regarding the size of the fusion pore, limiting measurements of the final stage of membrane fusion. In the present study, we used a biotinylated green fluorescence protein and streptavidin conjugated with Dylight 594 (DyStrp) as a Föster resonance energy transfer (FRET) donor and acceptor, respectively. This FRET pair encapsulated in each v-vesicle containing synaptobrevin and t-vesicle containing a binary acceptor complex of syntaxin 1a and synaptosomal-associated protein 25 revealed the opening of a large fusion pore of more than 5 nm, without the unwanted signals from unencapsulated dyes or leakage. This system enabled determination of the stoichiometry of the merging vesicles because the FRET efficiency of the FRET pair depended on the molar ratio between dyes. Here, we report a robust and informative assay for SNARE-mediated fusion pore opening. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Two dimensional laser induced fluorescence in the gas phase: a spectroscopic tool for studying molecular spectroscopy and dynamics

    Science.gov (United States)

    Gascooke, Jason R.; Lawrance, Warren D.

    2017-11-01

    Two dimensional laser induced fluorescence (2D-LIF) extends the usual laser induced fluorescence technique by adding a second dimension, the wavelength at which excited states emit, thereby significantly enhancing the information that can be extracted. It allows overlapping absorption features, whether they arise from within the same molecule or from different molecules in a mixture, to be associated with their appropriate "parent" state and/or molecule. While the first gas phase version of the technique was published a decade ago, the technique is in its infancy, having been exploited by only a few groups to date. However, its potential in gas phase spectroscopy and dynamics is significant. In this article we provide an overview of the technique and illustrate its potential with examples, with a focus on those utilising high resolution in the dispersed fluorescence dimension.

  20. Turn-on fluorescence probes based on pyranine/viologen charge-transfer complexes for the determination of nucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Schäferling, Michael, E-mail: Michael.schaeferling@utu.fi; Lang, Thomas; Schnettelker, Annette

    2014-10-15

    The formation of ground state charge-transfer complexes between pyranine (8-hydroxypyrene-1,3,6-trisulfonic acid) and viologen (paraquat) derivatives is utilized for the design of novel fluoroionophores for the determination of phosphate species, particularly of nucleotides. The strong quenching of the pyranine fluorescence by viologen-type charge transfer acceptors can be countermanded if these are functionalized with triethylammonium groups that serve as recognition elements for phosphate anions. We report on the fluorogenic responses of these water-soluble molecular probes in presence of different phosphates. Absorbance measurements give additional information on the charge transfer complex formation and the interaction with nucleotides. The experimental data show that these aggregates form attractive, simple and versatile fluorescence turn-on probes for nucleoside triphosphates. The reversibility of the fluorescence response is demonstrated by means of an enzymatic model assay using ATPase for the decomposition of adenosine triphosphate. - Highlights: • Pyranine/viologen charge-transfer complexes as molecular probe for ATP recognition. • Fluorescence turn on mechanism. • Selective compared to other nucleotides and phosphate anions. • Fast and reversible response applicable to monitor enzymatic reactions.

  1. Quinoline group based fluorescent sensor for detecting zinc ions in aqueous media and its logic gate behaviour

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Zhengping, E-mail: dongzhp@lzu.edu.cn [State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China); Guo, Yueping; Tian, Xin [State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China); Ma, Jiantai, E-mail: majiantai@lzu.edu.cn [State Key Laboratory of Applied Organic Chemistry, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China)

    2013-02-15

    A highly sensitive method for quantitative determination of Zn{sup 2+} in water has been developed by using a novel fluorescent sensor NQA: (N-Quinolin-8-yl-2-[(quinolin-8-ylcarbamoylmethyl)-amino]-acetamide). The sensor displays great selectivity for Zn{sup 2+} in the presence of other metal ions in aqueous solution and possesses an excellent sensitivity of about 2 Multiplication-Sign 10{sup -8} M for Zn{sup 2+}. The binding stoichiometry, binding affinity, and pH sensitivity of the sensor have also been studied. Furthermore, the fluorescent changes of NQA upon the addition of cations (Cu{sup 2+} and Zn{sup 2+}) are utilized to construct an INHIBIT logic gate at the molecular level, using Cu{sup 2+} and Zn{sup 2+} as chemical inputs and the fluorescence intensity as output. NQA has ideal chemical and spectroscopic properties that satisfy the criteria for further biological and environmental applications. - Highlights: Black-Right-Pointing-Pointer A novel fluorescent sensor for Zn{sup 2+} in water has been synthesized. Black-Right-Pointing-Pointer The sensor displays high selectivity for Zn{sup 2+} in the presence of other ions. Black-Right-Pointing-Pointer The sensor exhibits excellent sensing ability under the physiological pH window. Black-Right-Pointing-Pointer The sensor can be utilized as an INHIBIT logic gate at the molecular level.

  2. Sensitive Pb(2+) probe based on the fluorescence quenching by graphene oxide and enhancement of the leaching of gold nanoparticles.

    Science.gov (United States)

    Shi, Xinhao; Gu, Wei; Peng, Weidong; Li, Bingyu; Chen, Ningning; Zhao, Kai; Xian, Yuezhong

    2014-02-26

    A novel strategy was developed for fluorescent detection of Pb(2+) in aqueous solution based on the fact that graphene oxide (GO) could quench the fluorescence of amino pyrene (AP)-grafted gold nanoparticles (AP-AuNPs) and Pb(2+) could accelerate the leaching rate of AuNPs in the presence of S2O3(2-). In this system, fluorescence reporter AP was grafted on AuNPs through the Au-N bond. In the presence of GO, the system shows fluorescence quenching because of π-π stacking between AP and GO. With the addition of Pb(2+) and S2O3(2-), the system displays fluorescence recovery, which is attributed to the fact that Pb(2+) could accelerate the leaching of the AuNPs from GO surfaces and release of AP into aqueous solution. Interestingly, the concentration of GO could control the fluorescence "turn-off" or "turn-on" for Pb(2+) detection. In addition, GO is also an excellent promoter for the acceleration of the leaching of AuNPs and shortening the analytical time to ∼15 min. Under the optimal conditions, the fluorescence Pb(2+) sensor shows a linear range from 2.0 × 10(-9) to 2.3 × 10(-7) mol/L, with a detection limit of 1.0 × 10(-10) mol/L.

  3. The Synthesis of a Coumarin Carbohydrazide Dinuclear Copper Complex Based Fluorescence Probe and Its Detection of Thiols.

    Directory of Open Access Journals (Sweden)

    Guangjie He

    Full Text Available Small-molecule thiols, such as cysteine (CYS and glutathione (GSH, are essential for maintaining the cellular redox environment and play important roles in regulating various cellular physiological functions. A fluorescence probe (compound 1-Cu2+ for thiols based on coumarin carbohydrazide dinuclear copper complex was developed. Compound 1 was synthesized from the reaction of 7-(diethylamino-2-oxo-2H-chromene-3-carbohydrazide with 4-tert-butyl-2,6- diformylphenol. Accordingly, the copper complex (compound 1-Cu2+ was prepared by mixing compound 1 with 2 equivalents copper ions. Compound 1 had strong fluorescence while compound 1-Cu2+ hardly possessed fluorescence owing to the quenching nature of paramagnetism Cu2+ to the fluorescence molecule excited state. However, the fluorescence intensity of compound 1-Cu2+ was increased dramatically after the addition of thiol-containing amino acids, but not the other non-sulfhydryl amino acids. UV-vis absorption and fluorescence spectra indicated that compound 1-Cu2+ had good selectivity and sensitivity for thiols such as glutathione in CH3CN:H2O (3:2, v/v PBS solution. The fluorescence imaging experiments implied that compound 1-Cu2+ has potential application in thiol-containing amino acids detection in living cells.

  4. A wavelet-based Gaussian method for energy dispersive X-ray fluorescence spectrum

    Directory of Open Access Journals (Sweden)

    Pan Liu

    2017-05-01

    Full Text Available This paper presents a wavelet-based Gaussian method (WGM for the peak intensity estimation of energy dispersive X-ray fluorescence (EDXRF. The relationship between the parameters of Gaussian curve and the wavelet coefficients of Gaussian peak point is firstly established based on the Mexican hat wavelet. It is found that the Gaussian parameters can be accurately calculated by any two wavelet coefficients at the peak point which has to be known. This fact leads to a local Gaussian estimation method for spectral peaks, which estimates the Gaussian parameters based on the detail wavelet coefficients of Gaussian peak point. The proposed method is tested via simulated and measured spectra from an energy X-ray spectrometer, and compared with some existing methods. The results prove that the proposed method can directly estimate the peak intensity of EDXRF free from the background information, and also effectively distinguish overlap peaks in EDXRF spectrum.

  5. Efficient fluorescent deep-blue and hybrid white emitting devices based on carbazole/benzimidazole compound

    KAUST Repository

    Yang, Xiaohui

    2011-07-28

    We report the synthesis, photophysics, and electrochemical characterization of carbazole/benzimidazole-based compound (Cz-2pbb) and efficient fluorescent deep-blue light emitting devices based on Cz-2pbb with the peak external quantum efficiency of 4.1% and Commission Internationale dÉnclairage coordinates of (0.16, 0.05). Efficient deep-blue emission as well as high triplet state energy of Cz-2pbb enables fabrication of hybrid white organic light emitting diodes with a single emissive layer. Hybrid white emitting devices based on Cz-2pbb show the peak external quantum efficiency exceeding 10% and power efficiency of 14.8 lm/W at a luminance of 500 cd/m2. © 2011 American Chemical Society.

  6. Far-Red Fluorescent Probe for Imaging of Vicinal Dithiol-Containing Proteins in Living Cells Based on a pKa Shift Mechanism.

    Science.gov (United States)

    Zhang, Shengrui; Chen, Guojun; Wang, Yuanyuan; Wang, Qin; Zhong, Yaogang; Yang, Xiao-Feng; Li, Zheng; Li, Hua

    2018-02-20

    Vicinal dithiol-containing proteins (VDPs) play fundamental roles in intracellular redox homeostasis and are responsible for many diseases. In this work, we report a far-red fluorescence turn-on probe MCAs for VDPs exploiting the pK a shift of the imine functionality of the probe. MCAs is composed of a merocyanine Schiff base as the fluorescent reporter and a cyclic 1,3,2-dithiarsenolane as the specific ligand for VDPs. The imine pK a of MCAs is 4.8, and it exists predominantly in the Schiff base (SB) form at physiological pH. Due to the absence of a resonating positive charge, it absorbs at a relatively short wavelength and is essentially nonfluorescent. Upon selective binding to reduced bovine serum albumin (rBSA, selected as the model protein), MCAs was brought from aqueous media to the binding pockets of the protein, causing a large increase in pK a value of MCAs (pK a = 7.1). As a result, an increase in the protonated Schiff base (PSB) form of MCAs was observed at the physiological pH conditions, which in turn leads to a bathochromically shifted chromophore (λ abs = 634 nm) and a significant increase in fluorescence intensity (λ em = 657 nm) simultaneously. Furthermore, molecular dynamics simulations indicate that the salt bridges formed between the iminium in MCAs and the residues D72 and D517 in rBSA resist the dissociation of proton from the probe, thus inducing an increase of the pK a value. The proposed probe shows excellent sensitivity and specificity toward VDPs over other proteins and biologically relevant species and has been successfully applied for imaging of VDPs in living cells. We believe that the present pK a shift switching strategy may facilitate the development of new fluorescent probes that are useful for a wide range of applications.

  7. Development of Fluorescence Imaging Lidar for Boat-Based Coral Observation

    Directory of Open Access Journals (Sweden)

    Sasano Masahiko

    2016-01-01

    Full Text Available A fluorescence imaging lidar system installed in a boat-towable buoy has been developed for the observation of reef-building corals. Long-range fluorescent images of the sea bed can be recorded in the daytime with this system. The viability of corals is clear in these fluorescent images because of the innate fluorescent proteins. In this study, the specifications and performance of the system are shown.

  8. Development of a Protease Biosensor Based on a Dimerization-Dependent Red Fluorescent Protein.

    Science.gov (United States)

    Mitchell, Aaron C; Alford, Spencer C; Hunter, Sean A; Kannan, Deepti; Parra Sperberg, R Andres; Chang, Cheryl H; Cochran, Jennifer R

    2018-01-19

    Dysregulated activity of the protease matriptase is a key contributor to aggressive tumor growth, cancer metastasis, and osteoarthritis. Methods for the detection and quantification of matriptase activity and inhibition would be useful tools. To address this need, we developed a matriptase-sensitive protein biosensor based on a dimerization-dependent red fluorescent protein (ddRFP) reporter system. In this platform, two adjoining protein domains, connected by a protease-labile linker, produce fluorescence when assembled and are nonfluorescent when the linker is cleaved by matriptase. A panel of ddRFP-based matriptase biosensor designs was created that contained different linker lengths between the protein domains. These constructs were characterized for linker-specific cleavage, matriptase activity, and matriptase selectivity; a biosensor containing a RSKLRVGGH linker (termed B4) was expressed at high yields and displayed both high catalytic efficiency and matriptase specificity. This biosensor detects matriptase inhibition by soluble and yeast cell surface expressed inhibitor domains with up to a 5-fold dynamic range and also detects matriptase activity expressed by human cancer cell lines. In addition to matriptase, we highlight a strategy that can be used to create effective biosensors for quantifying activity and inhibition of other proteases of interest.

  9. Characterization of aluminum nitride based films with high resolution X-ray fluorescence spectroscopy

    Science.gov (United States)

    Anagnostopoulos, D. F.; Siozios, A.; Patsalas, P.

    2018-02-01

    X-ray fluorescence spectra of Al based films are measured, using a lab-scale wavelength dispersive flat crystal spectrometer. Various structures of AlN films were studied, like single layered, capped, stratified, nanostructured, crystalline, or amorphous. By optimizing the set-up for enhanced energy resolution and detection efficiency, the measured line shapes of Κα, Kβ, and KLL radiative Auger transitions are shown to be adequately detailed to allow chemical characterization. The chemistry identification is based on the pattern comparison of the emitted line shape from the chemically unknown film and the reference line shapes from standard materials, recorded under identical experimental conditions. The ultimate strength of lab-scale high resolution X-ray fluorescence spectroscopy on film analysis is verified, in cases that ordinary applied techniques like X-ray photoelectron and X-ray diffraction fail, while the characterization refers to the non-destructive determination of the bulk properties of the film and not to its surface, as the probed depth is in the micrometer range.

  10. Light-emitting diode and laser fluorescence-based devices in detecting occlusal caries

    Science.gov (United States)

    Rodrigues, Jonas A.; Hug, Isabel; Neuhaus, Klaus W.; Lussi, Adrian

    2011-10-01

    The aim of this study was to assess the performance of two light-emitting diode (LED)- and two laser fluorescence-based devices in detecting occlusal caries in vitro. Ninety-seven permanent molars were assessed twice by two examiners using two LED- (Midwest Caries - MID and VistaProof - VP) and two laser fluorescence-based (DIAGNOdent 2095 - LF and DIAGNOdent pen 2190 - LFpen) devices. After measuring, the teeth were histologically prepared and classified according to lesion extension. At D1 the specificities were 0.76 (LF and LFpen), 0.94 (MID), and 0.70 (VP); the sensitivities were 0.70 (LF), 0.62 (LFpen), 0.31 (MID), and 0.75 (VP). At D3 threshold the specificities were 0.88 (LF), 0.87 (LFpen), 0.90 (MID), and 0.70 (VP); the sensitivities were 0.63 (LF and LFpen), 0.70 (MID), and 0.96 (VP). Spearman's rank correlations with histology were 0.56 (LF), 0.51 (LFpen), 0.55 (MID), and 0.58 (VP). Inter- and intraexaminer ICC values were high and varied from 0.83 to 0.90. Both LF devices seemed to be useful auxiliary tools to the conventional methods, presenting good reproducibility and better accuracy at D3 threshold. MID was not able to differentiate sound surfaces from enamel caries and VP still needs improvement on the cut-off limits for its use.

  11. Quantifying Aflatoxin B1 in peanut oil using fabricating fluorescence probes based on upconversion nanoparticles

    Science.gov (United States)

    Sun, Cuicui; Li, Huanhuan; Koidis, Anastasios; Chen, Quansheng

    2016-08-01

    Rare earth doped upconversion nanoparticles convert near-infrared excitation light into visible emission light. Compared to organic fluorophores and semiconducting nanoparticles, upconversion nanoparticles (UCNPs) offer high photochemical stability, sharp emission bandwidths, and large anti-Stokes shifts. Along with the significant light penetration depth and the absence of autofluorescence in biological samples under infrared excitation, these UCNPs have attracted more and more attention on toxin detection and biological labelling. Herein, the fluorescence probe based on UCNPs was developed for quantifying Aflatoxin B1 (AFB1) in peanut oil. Based on a specific immunity format, the detection limit for AFB1 under optimal conditions was obtained as low as 0.2 ng·ml- 1, and in the effective detection range 0.2 to 100 ng·ml- 1, good relationship between fluorescence intensity and AFB1 concentration was achieved under the linear ratios up to 0.90. Moreover, to check the feasibility of these probes on AFB1 measurements in peanut oil, recovery tests have been carried out. A good accuracy rating (93.8%) was obtained in this study. Results showed that the nanoparticles can be successfully applied for sensing AFB1 in peanut oil.

  12. Label-free fluorescent detection of thrombin activity based on a recombinant enhanced green fluorescence protein and nickel ions immobilized nitrilotriacetic acid-coated magnetic nanoparticles.

    Science.gov (United States)

    Wang, Ming; Lei, Chunyang; Nie, Zhou; Guo, Manli; Huang, Yan; Yao, Shouzhuo

    2013-11-15

    Herein, a novel label-free fluorescent assay has been developed to detect the activity of thrombin and its inhibitor, based on a recombinant enhanced green fluorescence protein (EGFP) and Ni(2+) ions immobilized nitrilotriacetic acid-coated magnetic nanoparticles (Ni(2+)-NTA MNPs). The EGFP, containing a thrombin cleavage site and a hexahistidine sequence (His-tag) at its N-terminal, was adsorbed onto Ni(2+)-NTA MNPs through Ni(2+)-hexahistidine interaction, and dragged out of the solution by magnetic separation. Thrombin can selectively digest EGFP accompanied by His-tag peptide sequence leaving, and the resulting EGFP cannot be captured by Ni(2+)-NTA MNPs and kept in supernatant. Hence the fluorescence change of supernatant can clearly represent the activity of thrombin. Under optimized conditions, such assay showed a relatively low detection limit (3.0×10(-4) U mL(-1)), and was also used to detect the thrombin inhibitor, Hirudin, and further applied to detect thrombin activity in serum. Combined with the satisfactory reusability of Ni(2+)-NTA MNPs, our method presents a promising candidate for simple, sensitive, and cost-saving protease activity detecting and inhibitor screening. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Synchrotron radiation based micro X-ray fluorescence analysis of the calibration samples used in surface sensitive total reflection and grazing emission X-ray fluorescence techniques

    Science.gov (United States)

    Kubala-Kukuś, A.; Banaś, D.; Pajek, M.; Szlachetko, J.; Jagodziński, P.; Susini, J.; Salomé, M.

    2013-12-01

    Total reflection X-ray fluorescence (TXRF) and grazing emission X-ray fluorescence (GEXRF) are surface sensitive techniques and can be used for detailed surface studies of different materials, including ultra-low concentration contamination or the lateral and depth distributions of elements. The calibration procedure typically used involves placing a micro-droplet (˜μl) of the standard solution onto a silicon wafer (or quartz backing). After evaporation of the solvent, the residual amount of elements is used as a reference standard. Knowledge of the distribution of residue material on the substrate surface is crucial for precise quantification. In the present work the investigation of the lateral distribution of elements in the multielemental calibrating samples, containing the 23 most commonly studied elements, by using the synchrotron radiation based micro X-ray fluorescence is presented. The goal of this project was the study of a uniformity of the elemental distributions and determination of the residual elements morphology depending on the temperature of the drying process. The X-ray images were compared with optical and SEM images. Paper presents in details the experimental setup, sample preparation procedures, measurements and results. In the analysis of the X-ray images of the sample dried in high temperature the censoring approach was applied improving the quality of statistical analysis. The information on the elements distribution in the calibrating samples can be useful for developing more accurate calibration procedures applied in quantitative analysis of surface sensitive TXRF and GEXRF techniques.

  14. Boronic acids for fluorescence imaging of carbohydrates.

    Science.gov (United States)

    Sun, Xiaolong; Zhai, Wenlei; Fossey, John S; James, Tony D

    2016-02-28

    "Fluorescence imaging" is a particularly exciting and rapidly developing area of research; the annual number of publications in the area has increased ten-fold over the last decade. The rapid increase of interest in fluorescence imaging will necessitate the development of an increasing number of molecular receptors and binding agents in order to meet the demand in this rapidly expanding area. Carbohydrate biomarkers are particularly important targets for fluorescence imaging given their pivotal role in numerous important biological events, including the development and progression of many diseases. Therefore, the development of new fluorescent receptors and binding agents for carbohydrates is and will be increasing in demand. This review highlights the development of fluorescence imaging agents based on boronic acids a particularly promising class of receptors given their strong and selective binding with carbohydrates in aqueous media.

  15. BODIPY-based azamacrocyclic ensemble for selective fluorescence detection and quantification of homocysteine in biological applications.

    Science.gov (United States)

    Li, Zan; Geng, Zhi-Rong; Zhang, Cui; Wang, Xiao-Bo; Wang, Zhi-Lin

    2015-10-15

    Considering the significant role of plasma homocysteine in physiological processes, two ensembles (F465-Cu(2+) and F508-Cu(2+)) were constructed based on a BODIPY (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene) scaffold conjugated with an azamacrocyclic (1,4,7-triazacyclononane and 1,4,7,10-tetraazacyclododecane) Cu(2+) complex. The results of this effort demonstrated that the F465-Cu(2+) ensemble could be employed to detect homocysteine in the presence of other biologically relevant species, including cysteine and glutathione, under physiological conditions with high selectivity and sensitivity in the turn-on fluorescence mode, while the F508-Cu(2+) ensemble showed no fluorescence responses toward biothiols. A possible mechanism for this homocysteine-specific specificity involving the formation of a homocysteine-induced six-membered ring sandwich structure was proposed and confirmed for the first time by time-dependent fluorescence spectra, ESI-MS and EPR. The detection limit of homocysteine in deproteinized human serum was calculated to be 241.4 nM with a linear range of 0-90.0 μM and the detection limit of F465 for Cu(2+) is 74.7 nM with a linear range of 0-6.0 μM (F508, 80.2 nM, 0-7.0 μM). We have demonstrated the application of the F465-Cu(2+) ensemble for detecting homocysteine in human serum and monitoring the activity of cystathionine β-synthase in vitro. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Synthesis and properties of fluorescent hybrid nanocomposites based on copolyacrylates with dansyl semicarbazide units

    Energy Technology Data Exchange (ETDEWEB)

    Buruiana, Emil C., E-mail: emilbur@icmpp.r [' Petru Poni' Institute of Macromolecular Chemistry, 41A Grigore Ghica Voda Alley, 700487 Iasi (Romania); Chibac, Andreea L.; Buruiana, Tinca; Musteata, Valentina [' Petru Poni' Institute of Macromolecular Chemistry, 41A Grigore Ghica Voda Alley, 700487 Iasi (Romania)

    2011-07-15

    Our study examined a series of hybrid composites containing copolyacrylate with semicarbazide-dansyl groups prepared by conventional radical polymerization of monomers in the organic montmorillonite modified with alkyl chains of variable length or using the sol-gel technique. The structure and the chemical composition of the copolymers N-methacryloyloxyethylcarbamoyl-5- (dimethylaminonaphtalene-1-sulfonohydrazine)-co-methyl metahacrylate (DnsSA-co-MMA) and N-methacryloyloxyethylcarbamoyl -5-(dimethylaminonaphtalene-1-sulfonohydrazine)-co-dodecylacrylamide (DnsSA-co-DA) as well as their nanocomposites (HC-P1, HC-P2, HC-P3, HC-P4) were confirmed by spectral analysis ({sup 1}H NMR, FTIR, UV/vis), thermal methods and atomic force microscopy. To quantify the effect of the inorganic component compared to pure photopolymers we evaluated the properties of hybrid composites, including dielectric characterization. Additionally, these materials have been tested in experiments of fluorescence quenching by acids (HCl, p-toluenesulfonic acid, 1-S-camphorsulfonic acid), metallic cation (Cu{sup 2+}) and nitrobenzene. The results suggest that such nanocomposites could find applications as fluorescence-based chemosensors in homogeneous organic solutions or thin films. - Highlights: {yields} Dansylated hybrid composites were prepared by polymerization of monomers in organo-MMT or by sol-gel. {yields} Quenching effects by acids, Cu{sup 2+} and nitrobenzene in solution/film were evidenced. {yields} A fluorescence dequenching was observed for the composite with silsesquixane units. {yields} A reversible process occurs in the composite film exposed to nitrobenzene vapors.

  17. Unidirectional Light-Driven Molecular Motors Based on Overcrowded Alkenes

    NARCIS (Netherlands)

    Cnossen, Arjen; Browne, Wesley R.; Feringa, Ben L.; Credi, Alberto; Silvi, Serena; Venturi, Margherita

    2014-01-01

    Over the last two decades, interest in nanotechnology has led to the design and synthesis of a toolbox of nanoscale versions of macroscopic devices and components. In molecular nanotechnology, linear motors based on rotaxanes and rotary motors based on overcrowded alkenes are particularly promising

  18. Absorption and Fluorescence of PRODAN in Phospholipid Bilayers: A Combined Quantum Mechanics and Classical Molecular Dynamics Study

    Czech Academy of Sciences Publication Activity Database

    Cwiklik, Lukasz; Aquino, A. J. A.; Vazdar, Mario; Jurkiewicz, Piotr; Pittner, Jiří; Hof, Martin; Lischka, H.

    2011-01-01

    Roč. 115, č. 41 (2011), s. 11428-11437 ISSN 1089-5639 R&D Projects: GA AV ČR IAA400400810 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z40550506 Keywords : fluorescence * density functional theory * charge transfer Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.946, year: 2011

  19. Structure of the green fluorescent protein NowGFP with an anionic tryptophan-based chromophore.

    Science.gov (United States)

    Pletnev, Vladimir Z; Pletneva, Nadya V; Sarkisyan, Karen S; Mishin, Alexander S; Lukyanov, Konstantin A; Goryacheva, Ekaterina A; Ziganshin, Rustam H; Dauter, Zbigniew; Pletnev, Sergei

    2015-08-01

    A green-emitting fluorescent variant, NowGFP, with a tryptophan-based chromophore (Thr65-Trp66-Gly67) was recently developed from the cyan mCerulean by introducing 18 point mutations. NowGFP is characterized by bright green fluorescence at physiological and higher pH and by weak cyan fluorescence at low pH. Illumination with blue light induces irreversible photoconversion of NowGFP from a green-emitting to a cyan-emitting form. Here, the X-ray structures of intact NowGFP at pH 9.0 and pH 4.8 and of its photoconverted variant, NowGFP_conv, are reported at 1.35, 1.18 and 2.5 Å resolution, respectively. The structure of NowGFP at pH 9.0 suggests the anionic state of Trp66 of the chromophore to be the primary cause of its green fluorescence. At both examined pH values Trp66 predominantly adopted a cis conformation; only ∼ 20% of the trans conformation was observed at pH 4.8. It was shown that Lys61, which adopts two distinct pH-dependent conformations, is a key residue playing a central role in chromophore ionization. At high pH the side chain of Lys61 forms two hydrogen bonds, one to the indole N atom of Trp66 and the other to the carboxyl group of the catalytic Glu222, enabling an indirect noncovalent connection between them that in turn promotes Trp66 deprotonation. At low pH, the side chain of Lys61 is directed away from Trp66 and forms a hydrogen bond to Gln207. It has been shown that photoconversion of NowGFP is accompanied by decomposition of Lys61, with a predominant cleavage of its side chain at the C(γ)-C(δ) bond. Lys61, Glu222, Thr203 and Ser205 form a local hydrogen-bond network connected to the indole ring of the chromophore Trp66; mutation of any of these residues dramatically affects the spectral properties of NowGFP. On the other hand, an Ala150Val replacement in the vicinity of the chromophore indole ring resulted in a new advanced variant with a 2.5-fold improved photostability.

  20. A benzothiazole-based fluorescent probe for distinguishing and bioimaging of Hg{sup 2+} and Cu{sup 2+}

    Energy Technology Data Exchange (ETDEWEB)

    Gu, Biao; Huang, Liyan; Su, Wei; Duan, Xiaoli; Li, Haitao, E-mail: haitao-li@hunnu.edu.cn; Yao, Shouzhuo

    2017-02-15

    A new benzothiazole-based fluorescent probe 2-(benzo[d]thiazol-2-yl)-4-(1,3- dithian-2-yl)phenol (BT) with two different reaction sites, a thioacetal group (site 1 for Hg{sup 2+}), and O and N atoms of the benzothiazole dye (site 2 for Cu{sup 2+}), was designed and synthesized. The probe BT showed ratiometric fluorescent response to Hg{sup 2+} and fluorescence quenching behavior to Cu{sup 2+}, which induces naked-eye fluorescent color changes from green to blue and colorless, respectively. Moreover, it displayed highly sensitivity and selectivity toward Hg{sup 2+} and Cu{sup 2+} without interference from other metal ions. The sensing mechanisms were also confirmed by {sup 1}H NMR titration, mass spectrum and Job's plot analyses. Finally, probe BT was successfully used for fluorescent imaging of Hg{sup 2+} and Cu{sup 2+} in living cells, demonstrating its potential applications in biological science. - Highlights: • A benzothiazole-based probe for multiple metal ions has been firstly developed. • The differential sensing mechanisms of Hg{sup 2+} and Cu{sup 2+} relied on different reaction. • The probe could be used to monitor Hg{sup 2+} and Cu{sup 2+}in vitro and in vivo with distinct fluorescence changes.

  1. Introducing Ratiometric Fluorescence to MnO2Nanosheet-Based Biosensing: A Simple, Label-Free Ratiometric Fluorescent Sensor Programmed by Cascade Logic Circuit for Ultrasensitive GSH Detection.

    Science.gov (United States)

    Fan, Daoqing; Shang, Changshuai; Gu, Wenling; Wang, Erkang; Dong, Shaojun

    2017-08-09

    Glutathione (GSH) plays crucial roles in various biological functions, the level alterations of which have been linked to varieties of diseases. Herein, we for the first time expanded the application of oxidase-like property of MnO 2 nanosheet (MnO 2 NS) to fluorescent substrates of peroxidase. Different from previously reported fluorescent quenching phenomena, we found that MnO 2 NS could not only largely quench the fluorescence of highly fluorescent Scopoletin (SC) but also surprisingly enhance that of nonfluorescent Amplex Red (AR) via oxidation reaction. If MnO 2 NS is premixed with GSH, it will be reduced to Mn 2+ and lose the oxidase-like property, accompanied by subsequent increase in SC's fluorescence and decrease in AR's. On the basis of the above mechanism, we construct the first MnO 2 NS-based ratiometric fluorescent sensor for ultrasensitive and selective detection of GSH. Notably, this ratiometric sensor is programmed by the cascade logic circuit (an INHIBIT gate cascade with a 1 to 2 decoder). And a linear relationship between ratiometric fluorescent intensities of the two substrates and logarithmic values of GSH's concentrations is obtained. The detection limit of GSH is as low as 6.7 nM, which is much lower than previous ratiometric fluorescent sensors, and the lowest MnO 2 NS-based fluorescent GSH sensor reported so far. Furthermore, this sensor is simple, label-free, and low-cost; it also presents excellent applicability in human serum samples.

  2. Progress in molecular-based management of differentiated thyroid cancer

    Science.gov (United States)

    Xing, Mingzhao; Haugen, Bryan R; Schlumberger, Martin

    2014-01-01

    Substantial developments have occurred in the past 5–10 years in clinical translational research of thyroid cancer. Diagnostic molecular markers, such as RET-PTC, RAS, and BRAFV600E mutations; galectin 3; and a new gene expression classifier, are outstanding examples that have improved diagnosis of thyroid nodules. BRAF mutation is a prognostic genetic marker that has improved risk stratification and hence tailored management of patients with thyroid cancer, including those with conventionally low risks. Novel molecular-targeted treatments hold great promise for radioiodine-refractory and surgically inoperable thyroid cancers as shown in clinical trials; such treatments are likely to become a component of the standard treatment regimen for patients with thyroid cancer in the near future. These novel molecular-based management strategies for thyroid nodules and thyroid cancer are the most exciting developments in this unprecedented era of molecular thyroid-cancer medicine. PMID:23668556

  3. High-Capacitance Hybrid Supercapacitor Based on Multi-Colored Fluorescent Carbon-Dots.

    Science.gov (United States)

    Genc, Rukan; Alas, Melis Ozge; Harputlu, Ersan; Repp, Sergej; Kremer, Nora; Castellano, Mike; Colak, Suleyman Gokhan; Ocakoglu, Kasim; Erdem, Emre

    2017-09-11

    Multi-colored, water soluble fluorescent carbon nanodots (C-Dots) with quantum yield changing from 4.6 to 18.3% were synthesized in multi-gram using dated cola beverage through a simple thermal synthesis method and implemented as conductive and ion donating supercapacitor component. Various properties of C-Dots, including size, crystal structure, morphology and surface properties along with their Raman and electron paramagnetic resonance spectra were analyzed and compared by means of their fluorescence and electronic properties. α-Manganese Oxide-Polypyrrole (PPy) nanorods decorated with C-Dots were further conducted as anode materials in a supercapacitor. Reduced graphene oxide was used as cathode along with the dicationic bis-imidazolium based ionic liquid in order to enhance the charge transfer and wetting capacity of electrode surfaces. For this purpose, we used octyl-bis(3-methylimidazolium)diiodide (C8H16BImI) synthesized by N-alkylation reaction as liquid ionic membrane electrolyte. Paramagnetic resonance and impedance spectroscopy have been undertaken in order to understand the origin of the performance of hybrid capacitor in more depth. In particular, we obtained high capacitance value (C = 17.3 μF/cm 2 ) which is exceptionally related not only the quality of synthesis but also the choice of electrode and electrolyte materials. Moreover, each component used in the construction of the hybrid supercapacitor is also played a key role to achieve high capacitance value.

  4. Highly CO2 sensitive extruded fluorescent plastic indicator film based on HPTS.

    Science.gov (United States)

    Mills, Andrew; Yusufu, Dilidaer

    2016-02-07

    Highly-sensitive optical fluorescent extruded plastic films are reported for the detection of gaseous and dissolved CO2. The pH-sensitive fluorescent dye used is 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS, PTS(-)) coated on the surface of hydrophilic fumed silica and the base is tetrabutylammonium hydroxide (TBAH). The above components are used to create an HPTS pigment (i.e. HPTS/SiO2/TBAH) with a high CO2 sensitivity (%CO2 (S = 1/2) = 0.16%) and fast 50% response (t50↓) = 2 s and recovery (t50↑) = 5 s times. Highly CO2-sensitive plastic films are then fabricated, via the extrusion of the HPTS pigment powder in low-density polyethylene (LDPE). As with the HPTS-pigment, the luminescence intensity (at 515 nm) and absorbance (at 475 nm) of the HPTS plastic film decreases as the %CO2 in the ambient gas phase increases. The HPTS plastic film exhibits a high CO2 sensitivity, %CO2 (S = 1/2), of 0.29%, but a response time six month when stored in the dark but under otherwise ambient conditions). Moreover, the HPTS-LDPE film is stable in water, salt solution and even in acid (pH = 2), and in each of these media it can be used to detect dissolved CO2.

  5. Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell

    Science.gov (United States)

    Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2011-03-01

    A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.

  6. Medically Relevant Assays with a Simple Smartphone and Tablet Based Fluorescence Detection System

    Directory of Open Access Journals (Sweden)

    Piotr Wargocki

    2015-05-01

    Full Text Available Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 µg/mL and 0.938 µg in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis.

  7. Noninvasive and Quantitative Assessment of In Vivo Fetomaternal Interface Angiogenesis Using RGD-Based Fluorescence

    Directory of Open Access Journals (Sweden)

    M. Keramidas

    2014-01-01

    Full Text Available Angiogenesis is a key process for proper placental development and for the success of pregnancy. Although numerous in vitro methods have been developed for the assessment of this process, relatively few reliable in vivo methods are available to evaluate this activity throughout gestation. Here we report an in vivo technique that specifically measures placental neovascularization. The technique is based on the measurement of a fluorescent alpha v beta 3 (αvβ3 integrin-targeting molecule called Angiolone-Alexa-Fluor 700. The αvβ3 integrin is highly expressed by endothelial cells during the neovascularization and by trophoblast cells during their invasion of the maternal decidua. Angiolone was injected to gravid mice at 6.5 and 11.5 days post coitus (dpc. The fluorescence was analyzed one day later at 7.5 and 12.5 dpc, respectively. We demonstrated that (i Angiolone targets αvβ3 protein in the placenta with a strong specificity, (ii this technique is quantitative as the measurement was correlated to the increase of the placental size observed with increasing gestational age, and (iii information on the outcome is possible, as abnormal placentation could be detected early on during gestation. In conclusion, we report the validation of a new noninvasive and quantitative method to assess the placental angiogenic activity, in vivo.

  8. Construction of the Faraday Cup based on fluorescent screen as an electron beam sensor

    International Nuclear Information System (INIS)

    Sutadi; Rany Saptaaji; Suhartono; Sukaryono

    2016-01-01

    The Faraday Cup based on fluorescent screen as an electron beam profile sensor at electron accelerator has been conducted. In the principle, the electron beam which obtained from the electron source and accelerated in the accelerator tube will obtain the light which can be observed visually when it interact with fluorescent material (phosphorescent). This Faraday Cup for electron beam sensor was made from the modified TV tube. The main component of this Faraday Cup construction includes: 17 inch TV tube, SS reducer flange and the vacuum adhesive. There are two kind of test has been conducted, that is the vacuum level test and the electron beam sensor test. The vacuum level test was conducted by measuring the final vacuum level that can be reach, while the electron beam sensor test was conducted by monitoring of the electron beam profile that was trapped by Faraday Cup visually. The test result shows that TV tube can be modified as the Faraday Cup to sensor electron beam in the electron accelerator. (author)

  9. NitroxylFluor: A Thiol-Based Fluorescent Probe for Live-Cell Imaging of Nitroxyl.

    Science.gov (United States)

    Pino, Nicholas W; Davis, Jerome; Yu, Zhengxin; Chan, Jefferson

    2017-12-27

    Detection of nitroxyl (HNO), the transient one-electron reduced form of nitric oxide, is a significant challenge owing to its high reactivity with biological thiols (with rate constants as high as 10 9  M -1 s -1 ). To address this, we report a new thiol-based HNO-responsive trigger that can compete against reactive thiols for HNO. This process forms a common N-hydroxysulfenamide intermediate that cyclizes to release a masked fluorophore leading to fluorescence enhancement. To ensure that the cyclization step is rapid, our design capitalizes on two established physical organic phenomena; the alpha-effect and the Thorpe-Ingold effect. Using this new trigger, we developed NitroxylFluor, a selective HNO-responsive fluorescent probe. Treatment of NitroxylFluor with an HNO donor results in a 16-fold turn-on. This probe also exhibits excellent selectivity over various reactive nitrogen, oxygen, and sulfur species and efficacy in the presence of thiols (e.g., glutathione in mM concentrations). Lastly, we successfully performed live cell imaging of HNO using NitroxylFluor.

  10. Isotopic imaging via nuclear resonance fluorescence with laser-based Thomson radiation

    Science.gov (United States)

    Barty, Christopher P. J. [Hayward, CA; Hartemann, Frederic V [San Ramon, CA; McNabb, Dennis P [Alameda, CA; Pruet, Jason A [Brentwood, CA

    2009-07-21

    The present invention utilizes novel laser-based, high-brightness, high-spatial-resolution, pencil-beam sources of spectrally pure hard x-ray and gamma-ray radiation to induce resonant scattering in specific nuclei, i.e., nuclear resonance fluorescence. By monitoring such fluorescence as a function of beam position, it is possible to image in either two dimensions or three dimensions, the position and concentration of individual isotopes in a specific material configuration. Such methods of the present invention material identification, spatial resolution of material location and ability to locate and identify materials shielded by other materials, such as, for example, behind a lead wall. The foundation of the present invention is the generation of quasimonochromatic high-energy x-ray (100's of keV) and gamma-ray (greater than about 1 MeV) radiation via the collision of intense laser pulses from relativistic electrons. Such a process as utilized herein, i.e., Thomson scattering or inverse-Compton scattering, produces beams having diameters from about 1 micron to about 100 microns of high-energy photons with a bandwidth of .DELTA.E/E of approximately 10E.sup.-3.

  11. Quantification of total phosphorothioate in bacterial DNA by a bromoimane-based fluorescent method.

    Science.gov (United States)

    Xiao, Lu; Xiang, Yu

    2016-06-01

    The discovery of phosphorothioate (PT) modifications in bacterial DNA has challenged our understanding of conserved phosphodiester backbone structure of cellular DNA. This exclusive DNA modification in bacteria is not found in animal cells yet, and its biological function in bacteria is still poorly understood. Quantitative information about the bacterial PT modifications is thus important for the investigation of their possible biological functions. In this study, we have developed a simple fluorescence method for selective quantification of total PTs in bacterial DNA, based on fluorescent labeling of PTs and subsequent release of the labeled fluorophores for absolute quantification. The method was highly selective to PTs and not interfered by the presence of reactive small molecules or proteins. The quantification of PTs in an E. coli DNA sample was successfully achieved using our method and gave a result of about 455 PTs per million DNA nucleotides, while almost no detectable PTs were found in a mammalian calf thymus DNA. With this new method, the content of phosphorothioate in bacterial DNA could be successfully quantified, serving as a simple method suitable for routine use in biological phosphorothioate related studies. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Fluorescence array-based sensing of metal ions using conjugated polyelectrolytes.

    Science.gov (United States)

    Wu, Yi; Tan, Ying; Wu, Jiatao; Chen, Shangying; Chen, Yu Zong; Zhou, Xinwen; Jiang, Yuyang; Tan, Chunyan

    2015-04-01

    Array-based sensing offers several advantages for detecting a series of analytes with common structures or properties. In this study, four anionic conjugated polyelectrolytes (CPEs) with a common poly(p-pheynylene ethynylene) (PPE) backbone and varying pendant ionic side chains were designed. The conjugation length, repeat unit pattern, and ionic side chain composition were the main factors affecting the fluorescence patterns of CPE polymers in response to the addition of different metal ions. Eight metal ions, including Pb(2+), Hg(2+), Fe(3+), Cr(3+), Cu(2+), Mn(2+), Ni(2+), and Co(2+), categorized as water contaminants by the Environmental Protection Agency, were selected as analytes in this study. Fluorescence intensity response patterns of the four-PPE sensor array toward each of the metal ions were recorded, analyzed, and transformed into canonical scores using linear discrimination analysis (LDA), which permitted clear differentiation between metal ions using both two-dimensional and three-dimensional graphs. In particular, the array could readily differentiate between eight toxic metal ions in separate aqueous solutions at 100 nM. Our four-PPE sensor array also provides a practical application to quantify Pb(2+) and Hg(2+) concentrations in blind samples within a specific concentration range.

  13. Isolation and selection of fluorescent pseudomonads based on multiple plant growth promotion traits and siderotyping

    Directory of Open Access Journals (Sweden)

    Jayamohan Subramanian

    2014-09-01

    Full Text Available Fluorescent pseudomonads, acclaimed plant associated bacterial group, are well-known plant growth promoting-biocontrol agents in rhizosphere arena. In this study, 144 fluorescent pseudomonad isolates from rhizosphere soil samples were screened with King's medium B supplemented with 8-hydroxyquinoline (8-HQ chelator and comprehensively profiled for plant growth promotion viz., production of indole acetic acid (IAA, siderophore, ammonia, hydrogen cyanide, motility, phosphate solubilization, root growth promotion, and biofilm forming ability, along with two known control strains of pseudomonads. Iron and IAA regulated secondary metabolite siderophore production were investigated quantitatively. All isolates were positive for ammonia production and motility; 46% isolates were positive for hydrogen cyanide, 44% shown positivity for phosphate solubilization, and 40% isolates for siderophore production. Siderotyping showed production of hydroxamate type of siderophores which are known to be more efficient biocontrol agents. All isolates stimulated root growth to varying extent and had potentiality to form biofilms, a critical constituent for survival on different environments. Forty-two isolates of pseudomonads showed antagonistic behavior against the deleterious fungal pathogen Fusarium oxysporum (MTCC1755. Based on the above observations and statistical analysis, 11 isolates were shortlisted for further scrutiny. The study of biogeographic correlation and secondary metabolite profiling in association with plant growth promotion focalizes significant assessment on the behavior and antagonistic action, which probably brings out a competent biocontrol agent in a sustainable eco-friendly dimension.

  14. Fiber-based time-resolved fluorescence and phosphorescence spectroscopy of tumors

    Science.gov (United States)

    Shirmanova, M.; Lukina, M.; Orlova, A.; Studier, H.; Zagaynova, E.; Becker, W.; Shcheslavskiy, V.

    2017-07-01

    The study of metabolic and oxygen states of cells in a tumor in vivo is crucial for understanding of the mechanisms responsible for the tumor development and provides background for the relevant tumor's treatment. Here, we show that a specially designed implantable fiber-optical probe provides a promising tool for optical interrogation of metabolic and oxygen states of a tumor in vivo. In our experiments, the excitation light from a ps diode laser source is delivered to the sample through an exchangeable tip via a multimode fiber, and the emission light is transferred to the detector by another multimode fiber. Fluorescence lifetime of nicotinamid adenine dinucleotide (NAD(P)H) and phosphorescence lifetime of an oxygen sensor based on iridium (III) complex of enzothienylpyridine (BTPDM1) are explored both in model experiment in solutions, and in living mice. The luminescence spectroscopy data is substantiated with immunohistochemistry experiments. To the best of our knowledge, the measurements of both metabolic status and oxygenation of tumor in vivo by fluorescence/phosphorescence lifetime spectroscopy with a fiber-optic probe are done for the first time.

  15. Fluorescence interference contrast based approach to study real time interaction of melittin with plasma membranes

    Science.gov (United States)

    Gupta, Sharad; Gui, Dong; Zandi, Roya; Gill, Sarjeet; Mohideen, Umar

    2014-03-01

    Melittin is an anti-bacterial and hemolytic toxic peptide found in bee venom. Cell lysis behavior of peptides has been widely investigated, but the exact interaction mechanism of lytic peptides with lipid membranes and its constituents has not been understood completely. In this paper we study the melittin interaction with lipid plasma membranes in real time using non-invasive and non-contact fluorescence interference contrast microscopy (FLIC). Particularly the interaction of melittin with plasma membranes was studied in a controlled molecular environment, where these plasma membrane were composed of saturated lipid, 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and unsaturated lipid, 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC) with and without cholesterol. We found out that melittin starts to form nanometer size pores in the plasma membranes shortly after interacting with membranes. But the addition of cholesterol in plasma membrane slows down the pore formation process. Our results show that inclusion of cholesterol to the plasma membranes make them more resilient towards pore formation and lysis of membrane.

  16. Wide-field time-domain fluorescence lifetime imaging microscopy (FLIM): Molecular snapshots of metabolic function in biological systems

    Science.gov (United States)

    Sud, Dhruv

    2008-12-01

    Steady-state fluorescence imaging is routinely employed to obtain physiological information but is susceptible to artifacts such as absorption and photobleaching. FLIM provides an additional source of contrast oblivious to these but is affected by factors such as pH, gases, and temperature. Here we focused on developing a resolution-enhanced FLIM system for quantitative oxygen sensing. Oxygen is one of the most critical components of metabolic machinery and affects growth, differentiation, and death. FLIM-based oxygen sensing provides a valuable tool for biologists without the need of alternate technologies. We also developed novel computational approaches to improve spatial resolution of FLIM images, extending its potential for thick tissue studies. We designed a wide-field time-domain UV-vis-NIR FLIM system with high temporal resolution (50 ps), large temporal dynamic range (750 ps -- 1 mus), short data acquisition/processing times (15 s) and noise-removal capability. Lifetime calibration of an oxygen-sensitive, ruthenium dye (RTDP) enabled in vivo oxygen level measurements (resolution = 8 muM, range = 1 -- 300 muM). Combining oxygen sensing with endogenous imaging allowed for the study of two key molecules (NADH and oxygen) consumed at the termini of the oxidative phosphorylation pathway in Barrett's adenocarcinoma columnar (SEG-1) cells and Esophageal normal squamous cells (HET-1). Starkly higher intracellular oxygen and NADH levels in living SEG-1 vs. HET-1 cells were detected by FLIM and attributed to altered metabolic pathways in malignant cells. We performed FLIM studies in microfluidic bioreactors seeded with mouse myoblasts. For these systems, oxygen concentrations play an important role in cell behavior and gene expression. Oxygen levels decreased with increasing cell densities and were consistent with simulated model outcomes. In single bioreactor loops, FLIM detected spatial heterogeneity in oxygen levels as high as 20%. We validated our calibration

  17. Graphitic Carbon Nitride Nanosheets-Based Ratiometric Fluorescent Probe for Highly Sensitive Detection of H2O2 and Glucose.

    Science.gov (United States)

    Liu, Jin-Wen; Luo, Ying; Wang, Yu-Min; Duan, Lu-Ying; Jiang, Jian-Hui; Yu, Ru-Qin

    2016-12-14

    Graphitic carbon nitride (g-C 3 N 4 ) nanosheets, an emerging graphene-like carbon-based nanomaterial with high fluorescence and large specific surface areas, hold great potential for biosensor applications. Current g-C 3 N 4 nanosheets based fluorescent biosensors majorly rely on single fluorescent intensity reading through fluorescence quenching interactions between the nanosheets and metal ions. Here we report for the first time the development of a novel g-C 3 N 4 nanosheets-based ratiometric fluorescence sensing strategy for highly sensitive detection of H 2 O 2 and glucose. With o-phenylenediamine (OPD) oxidized by H 2 O 2 in the presence of horseradish peroxidase (HRP), the oxidization product can assemble on the g-C 3 N 4 nanosheets through hydrogen bonding and π-π stacking, which effectively quenches the fluorescence of g-C 3 N 4 while delivering a new emission peak. The ratiometric signal variations enable robust and sensitive detection of H 2 O 2 . On the basis of the glucose converting into H 2 O 2 through the catalysis of glucose oxidase, the g-C 3 N 4 -based ratiometric fluorescence sensing platform is also exploited for glucose assay. The developed strategy is demonstrated to give a detection limit of 50 nM for H 2 O 2 and 0.4 μM for glucose, at the same time, it has been successfully used for glucose levels detection in human serum. This strategy may provide a cost-efficient, robust, and high-throughput platform for detecting various species involving H 2 O 2 -generation reactions for biomedical applications.

  18. Context based mixture model for cell phase identification in automated fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Zhou Xiaobo

    2007-01-01

    Full Text Available Abstract Background Automated identification of cell cycle phases of individual live cells in a large population captured via automated fluorescence microscopy technique is important for cancer drug discovery and cell cycle studies. Time-lapse fluorescence microscopy images provide an important method to study the cell cycle process under different conditions of perturbation. Existing methods are limited in dealing with such time-lapse data sets while manual analysis is not feasible. This paper presents statistical data analysis and statistical pattern recognition to perform this task. Results The data is generated from Hela H2B GFP cells imaged during a 2-day period with images acquired 15 minutes apart using an automated time-lapse fluorescence microscopy. The patterns are described with four kinds of features, including twelve general features, Haralick texture features, Zernike moment features, and wavelet features. To generate a new set of features with more discriminate power, the commonly used feature reduction techniques are used, which include Principle Component Analysis (PCA, Linear Discriminant Analysis (LDA, Maximum Margin Criterion (MMC, Stepwise Discriminate Analysis based Feature Selection (SDAFS, and Genetic Algorithm based Feature Selection (GAFS. Then, we propose a Context Based Mixture Model (CBMM for dealing with the time-series cell sequence information and compare it to other traditional classifiers: Support Vector Machine (SVM, Neural Network (NN, and K-Nearest Neighbor (KNN. Being a standard practice in machine learning, we systematically compare the performance of a number of common feature reduc