photobleaching

Summary

Summary: Light-induced change in a chromophore, resulting in the loss of its absorption of light of a particular wave length. The photon energy causes a conformational change in the photoreceptor proteins affecting PHOTOTRANSDUCTION. This occurs naturally in the retina (ADAPTATION, OCULAR) on long exposure to bright light. Photobleaching presents problems when occurring in PHOTODYNAMIC THERAPY, and in FLUORESCENCE MICROSCOPY. On the other hand, this phenomenon is exploited in the technique, FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING, allowing measurement of the movements of proteins and LIPIDS in the CELL MEMBRANE.

Top Publications

  1. ncbi Bright fluorescent nanodiamonds: no photobleaching and low cytotoxicity
    Shu Jung Yu
    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan 106, ROC
    J Am Chem Soc 127:17604-5. 2005
  2. ncbi Imaging intracellular fluorescent proteins at nanometer resolution
    Eric Betzig
    Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA
    Science 313:1642-5. 2006
  3. ncbi Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching
    Cindi L Schwartz
    Boulder Laboratory for 3D Electron Microscopy of Cells, University of Colorado, Department of Molecular, Cellular, and Developmental Biology, Boulder, CO, USA
    J Microsc 227:98-109. 2007
  4. ncbi On the mechanism of Trolox as antiblinking and antibleaching reagent
    Thorben Cordes
    Angewandte Physik Biophysik, and Center for NanoScience, Ludwig Maximilians Universitat, Amalienstrasse 54, 80799 Munchen, Germany
    J Am Chem Soc 131:5018-9. 2009
  5. ncbi A spectroscopic study of the photobleaching of protoporphyrin IX in solution
    M B Ericson
    Department of Experimental Physics, School of Physics and Engineering Physics, Chalmers University of Technology, Goteborg University, SE 412 96 Goteborg, Sweden
    Lasers Med Sci 18:56-62. 2003
  6. pmc Cross-validating FRAP and FCS to quantify the impact of photobleaching on in vivo binding estimates
    Timothy J Stasevich
    National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 99:3093-101. 2010
  7. ncbi Histone dynamics in living cells revealed by photobleaching
    Hiroshi Kimura
    Nuclear Function and Dynamics Unit, HMRO, School of Medicine, Kyoto University, Yoshida konoe cho, Sakyo ku, Kyoto 606 8501, Japan
    DNA Repair (Amst) 4:939-50. 2005
  8. ncbi Reversible photobleaching of enhanced green fluorescent proteins
    Daniel Sinnecker
    Institut fur Pharmakologie, Charite Universitatsmedizin Berlin, Campus Benjamin Franklin, Thielallee 67 73, 14195 Berlin, Germany
    Biochemistry 44:7085-94. 2005
  9. ncbi Photobleaching kinetics, photoproduct formation, and dose estimation during ALA induced PpIX PDT of MLL cells under well oxygenated and hypoxic conditions
    Jonathan S Dysart
    Juravinski Cancer Centre, Physics Research, 699 Concession St, Hamilton, Ontario, Canada
    Photochem Photobiol Sci 5:73-81. 2006
  10. pmc Analysis of red-fluorescent proteins provides insight into dark-state conversion and photodegradation
    Kevin M Dean
    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado, USA
    Biophys J 101:961-9. 2011

Detail Information

Publications269 found, 100 shown here

  1. ncbi Bright fluorescent nanodiamonds: no photobleaching and low cytotoxicity
    Shu Jung Yu
    Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan 106, ROC
    J Am Chem Soc 127:17604-5. 2005
    ..The fluorescent nanodiamonds (FND) show no sign of photobleaching and can be taken up by mammalian cells with minimal cytotoxicity...
  2. ncbi Imaging intracellular fluorescent proteins at nanometer resolution
    Eric Betzig
    Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA
    Science 313:1642-5. 2006
    ....
  3. ncbi Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching
    Cindi L Schwartz
    Boulder Laboratory for 3D Electron Microscopy of Cells, University of Colorado, Department of Molecular, Cellular, and Developmental Biology, Boulder, CO, USA
    J Microsc 227:98-109. 2007
    ..Once located, the sample can be placed in the EM for further study at higher resolution. An additional benefit of the cryo-LM stage is that photobleaching is slower at cryogenic temperatures (-140 degrees C) than at room temperature.
  4. ncbi On the mechanism of Trolox as antiblinking and antibleaching reagent
    Thorben Cordes
    Angewandte Physik Biophysik, and Center for NanoScience, Ludwig Maximilians Universitat, Amalienstrasse 54, 80799 Munchen, Germany
    J Am Chem Soc 131:5018-9. 2009
    Recent advances in photobleaching and blinking prevention have aided the advancement of single-molecule and super-resolution fluorescence microscopy...
  5. ncbi A spectroscopic study of the photobleaching of protoporphyrin IX in solution
    M B Ericson
    Department of Experimental Physics, School of Physics and Engineering Physics, Chalmers University of Technology, Goteborg University, SE 412 96 Goteborg, Sweden
    Lasers Med Sci 18:56-62. 2003
    ..In this study, the involvement of oxygen in the photobleaching process was verified by studying the effect of oxygen depletion using the freeze-pump-thaw (FPT) method...
  6. pmc Cross-validating FRAP and FCS to quantify the impact of photobleaching on in vivo binding estimates
    Timothy J Stasevich
    National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 99:3093-101. 2010
    ..To address this uncertainty, we compare fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) measurements of the binding kinetics of a transcription ..
  7. ncbi Histone dynamics in living cells revealed by photobleaching
    Hiroshi Kimura
    Nuclear Function and Dynamics Unit, HMRO, School of Medicine, Kyoto University, Yoshida konoe cho, Sakyo ku, Kyoto 606 8501, Japan
    DNA Repair (Amst) 4:939-50. 2005
    ..Particularly, photobleaching techniques like fluorescence recovery after photobleaching (FRAP) revealed the mobility of many nuclear ..
  8. ncbi Reversible photobleaching of enhanced green fluorescent proteins
    Daniel Sinnecker
    Institut fur Pharmakologie, Charite Universitatsmedizin Berlin, Campus Benjamin Franklin, Thielallee 67 73, 14195 Berlin, Germany
    Biochemistry 44:7085-94. 2005
    ..used for multicolor imaging, fluorescence resonance energy transfer (FRET), and fluorescence recovery after photobleaching (FRAP)...
  9. ncbi Photobleaching kinetics, photoproduct formation, and dose estimation during ALA induced PpIX PDT of MLL cells under well oxygenated and hypoxic conditions
    Jonathan S Dysart
    Juravinski Cancer Centre, Physics Research, 699 Concession St, Hamilton, Ontario, Canada
    Photochem Photobiol Sci 5:73-81. 2006
    Fluorescence photobleaching and photoproduct formation were investigated during delta-aminolevulinic acid (ALA) induced protoporphyrin IX (PpIX) PDT of MLL cells in vitro. Cells were incubated in either 0.1 or 1...
  10. pmc Analysis of red-fluorescent proteins provides insight into dark-state conversion and photodegradation
    Kevin M Dean
    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado, USA
    Biophys J 101:961-9. 2011
    ..However, the brightness of FPs is limited by dark-state conversion (DSC) and irreversible photobleaching, which occur on different timescales...
  11. ncbi Coexistence of fast photodarkening and slow photobleaching in Ge19As21Se60 thin films
    Pritam Khan
    Department of Physics, Indian Institute of Science Education and Research, Bhopal 462023, India
    Opt Express 20:12416-21. 2012
    ..fast photodarkening (PD) and slow photobleaching (PB) in Ge(19)As(21)Se(60) thin films, when illuminated with a laser of wavelength 671 nm...
  12. pmc Structural basis for reversible photobleaching of a green fluorescent protein homologue
    J Nathan Henderson
    Departments of Chemistry and Physics, and Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA
    Proc Natl Acad Sci U S A 104:6672-7. 2007
    ..The fundamental mechanism appears to be common to all of the photoactivatable and reversibly photoswitchable FPs reported to date...
  13. pmc Quantitative fluorescence loss in photobleaching for analysis of protein transport and aggregation
    Daniel Wüstner
    Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M, DK 5230, Denmark
    BMC Bioinformatics 13:296. 2012
    Fluorescence loss in photobleaching (FLIP) is a widely used imaging technique, which provides information about protein dynamics in various cellular regions...
  14. ncbi In vivo measurement of brain extracellular space diffusion by cortical surface photobleaching
    Devin K Binder
    Department of Medicine, Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California 94143 0521, USA
    J Neurosci 24:8049-56. 2004
    ..We developed a brain surface photobleaching method to measure the diffusion of fluorescently labeled macromolecules in the ECS of the cerebral cortex...
  15. pmc Protein-protein interactions between lens vimentin and alphaB-crystallin using FRET acceptor photobleaching
    Shuhua Song
    Center for Ophthalmic Research Surgery, Brigham and Women s Hospital, Boston, MA 02115, USA
    Mol Vis 14:1282-7. 2008
    ..We hypothesize that alteration of protein-protein interaction between R120G alphaB-crystallin and lens intermediate filament proteins is one of the mechanisms of congenital cataract...
  16. ncbi Electron tomography of the Maurer's cleft organelles of Plasmodium falciparum-infected erythrocytes reveals novel structural features
    Eric Hanssen
    Department of Biochemistry and Centre of Excellence for Coherent X ray Science, La Troube University, Melbourne, Victoria 3086, Australia
    Mol Microbiol 67:703-18. 2008
    ..Spot fluorescence photobleaching was used to demonstrate the absence of a lipid continuum between the Maurer's clefts and parasite membranes ..
  17. pmc Determination of the oligomer size of amyloidogenic protein beta-amyloid(1-40) by single-molecule spectroscopy
    Hao Ding
    Biophysics Research Division, University of Michigan, Ann Arbor, Michigan, USA
    Biophys J 97:912-21. 2009
    ..We find that by directly counting the photobleaching steps in the fluorescence, we can determine the number of subunits in individual beta-amyloid(1-40) oligomers, ..
  18. pmc Heat stress causes inhibition of the de novo synthesis of antenna proteins and photobleaching in cultured Symbiodinium
    Shunichi Takahashi
    Australian Research Council Center of Excellence in Plant Energy Biology, Research School of Biological Sciences, Australian National University, Australian Capital Territory, Canberra, Australia
    Proc Natl Acad Sci U S A 105:4203-8. 2008
    ..Here, we show that moderate heat stress causes photobleaching through inhibition of the de novo synthesis of intrinsic light-harvesting antennae [chlorophyll a-chlorophyll ..
  19. ncbi Sensitivity of CFP/YFP and GFP/mCherry pairs to donor photobleaching on FRET determination by fluorescence lifetime imaging microscopy in living cells
    Marc Tramier
    Institut Jacques Monod, UMR 7592 CNRS University Paris 6 University Paris 7, 2 place Jussieu, 75251 Paris Cedex 05, France
    Microsc Res Tech 69:933-9. 2006
    ..It can be difficult, however, to avoid photobleaching when observing fluorescent cells under the microscope, especially those expressing CFP...
  20. ncbi Microfiberoptic fluorescence photobleaching reveals size-dependent macromolecule diffusion in extracellular space deep in brain
    Zsolt Zador
    Department of Medicine, University of California, San Francisco, CA, USA
    FASEB J 22:870-9. 2008
    ..diffusion in normally light-inaccessible regions of mouse brain by microfiberoptic epifluorescence photobleaching, in which a fiberoptic with a micron-size tip is introduced deep in brain tissue...
  21. ncbi Fixation, mounting and sealing with nail polish of cell specimens lead to incorrect FRET measurements using acceptor photobleaching
    Simona Rodighiero
    CIMAINA, Salzburg, Austria
    Cell Physiol Biochem 21:489-98. 2008
    ..of the FRET efficiency by de-quenching the donor cyan-fluorescent-protein (CFP) emission due to acceptor-photobleaching leads to erroneous estimate of the FRET efficiency in fixed, mounted and sealed specimens...
  22. ncbi Photobleaching of reduced nicotinamide adenine dinucleotide and the development of highly fluorescent lesions in rat basophilic leukemia cells during multiphoton microscopy
    LeAnn M Tiede
    Department of Biomedical Sciences, Creighton University, Omaha, NE, USA
    Photochem Photobiol 82:656-64. 2006
    ..provides an intrinsic indicator of the cellular metabolic state, but prolonged monitoring is limited by photobleaching and/or phototoxicity...
  23. ncbi High-precision FLIM-FRET in fixed and living cells reveals heterogeneity in a simple CFP-YFP fusion protein
    Michael Millington
    Collaborative Optical Spectroscopy, Micromanipulation and Imaging Centre COSMIC, The University of Edinburgh, King s Buildings, Edinburgh EH9 3JZ, UK
    Biophys Chem 127:155-64. 2007
    ..Our results for this simple control system indicate that the in vivo FLIM-FRET studies of more complex protein-protein interactions would benefit greatly from such quantitative measurements...
  24. pmc Quantifying transient binding of ISWI chromatin remodelers in living cells by pixel-wise photobleaching profile evolution analysis
    Fabian Erdel
    Research Group Genome Organization and Function, Deutsches Krebsforschungszentrum DKFZ and BioQuant, 69120 Heidelberg, Germany
    Proc Natl Acad Sci U S A 109:E3221-30. 2012
    ..To reach this goal, we have developed a fluorescence bleaching technique termed pixel-wise photobleaching profile evolution analysis (3PEA)...
  25. pmc Photobleaching response of different sources of chromophoric dissolved organic matter exposed to natural solar radiation using absorption and excitation-emission matrix spectra
    Yunlin Zhang
    Taihu Lake Laboratory Ecosystem Research Station, State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing, China
    PLoS ONE 8:e77515. 2013
    ..cycle is driven by several physical and biological processes such as river input, biogeneration and photobleaching that act as primary sinks and sources of CDOM...
  26. ncbi Characterization of Photofrin photobleaching for singlet oxygen dose estimation during photodynamic therapy of MLL cells in vitro
    Jonathan S Dysart
    Juravinski Cancer Centre, Physics Research, 699 Concession St, Hamilton, Ontario L8V 5C2, Canada
    Phys Med Biol 50:2597-616. 2005
    A singlet oxygen dose model is developed for PDT with Photofrin. The model is based on photosensitizer photobleaching kinetics, and incorporates both singlet oxygen and non-singlet oxygen mediated bleaching mechanisms...
  27. ncbi Structural basis of X-ray-induced transient photobleaching in a photoactivatable green fluorescent protein
    Virgile Adam
    European Synchrotron Radiation Facility, 6 rue Jules Horowitz, BP 220, 38043 Grenoble Cedex, France
    J Am Chem Soc 131:18063-5. 2009
    ..that the methylene bridge of the chromophore is the Achilles' heel of fluorescent proteins and help unravel the mechanisms of blinking and photobleaching in FPs, which are of importance in the rational design of photostable variants.
  28. pmc Simulations of measured photobleaching kinetics in human basal cell carcinomas suggest blood flow reductions during ALA-PDT
    Ken Kang Hsin Wang
    Department of Physics and Astronomy, University of Rochester, Rochester, New York 14627, USA
    Lasers Surg Med 41:686-96. 2009
    ..Protoporphyrin IX (PpIX) photobleaching in the lesion and the adjacent perilesion normal margin was monitored by fluorescence spectroscopy...
  29. ncbi On the origin of the optical properties of humic substances
    Rossana Del Vecchio
    Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA
    Environ Sci Technol 38:3885-91. 2004
    Absorption and fluorescence spectroscopy and laser photobleaching experiments were employed to probe the origins of the optical properties of humic substances (HS)...
  30. pmc Investigation of stoichiometry of T4 bacteriophage helicase loader protein (gp59)
    Sri Ranjini Arumugam
    Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA
    J Biol Chem 284:29283-9. 2009
    ..In this study, we employed single-molecule photobleaching (smPB) experiments to elucidate the stoichiometry of gp59 on a forked DNA and to investigate its interaction ..
  31. pmc Inferring subunit stoichiometry from single molecule photobleaching
    Keegan E Hines
    Section of Neurobiology, The University of Texas at Austin, Austin, TX 78712, USA
    J Gen Physiol 141:737-46. 2013
    Single molecule photobleaching is a powerful tool for determining the stoichiometry of protein complexes...
  32. ncbi Photobleaching with phloxine B sensitizer to reduce food matrix interference for detection of Escherichia coli serotype O157:H7 in fresh spinach by flow cytometry
    Dan A Buzatu
    U S Food and Drug Administration, National Center for Toxicological Research, Jefferson, AR 72079, USA
    Food Microbiol 36:416-25. 2013
    ..The RAPID-B method limit of detection (LOD) was one cell in 25 g. It proved superior to the reference method (which incorporated real time-PCR, selective enrichment, and culture plating elements) in accuracy and speed...
  33. ncbi Localization of peroxisomal matrix proteins by photobleaching
    Charlotta Buch
    Department of Biosciences and Nutrition, Karolinska Institutet, SE 141 57 Huddinge, Sweden
    Biochem Biophys Res Commun 388:355-9. 2009
    ..We have used photobleaching in live cells expressing green fluorescent protein (GFP)-fusion proteins to show that imported bona fide ..
  34. ncbi Structural evidence for a two-regime photobleaching mechanism in a reversibly switchable fluorescent protein
    Chenxi Duan
    Université Grenoble Alpes, Institut de Biologie Structurale IBS, F 38027 Grenoble, France
    J Am Chem Soc 135:15841-50. 2013
    b>Photobleaching, the irreversible photodestruction of a chromophore, severely limits the use of fluorescent proteins (FPs) in optical microscopy. Yet, the mechanisms that govern photobleaching remain poorly understood...
  35. ncbi Photodynamic therapy of actinic keratosis at varying fluence rates: assessment of photobleaching, pain and primary clinical outcome
    M B Ericson
    Department of Dermatology, Sahlgrenska University Hospital, Goteborg University, SE 413 45 Goteborg, Sweden
    Br J Dermatol 151:1204-12. 2004
    ..Recent studies in rodents have shown that a low fluence rate is preferable due to depletion of oxygen at high fluence rates. However, these results have not yet been verified in humans...
  36. pmc Measuring the stoichiometry of functional PspA complexes in living bacterial cells by single molecule photobleaching
    Tchern Lenn
    Division of Biology, Imperial College London, London, UK SW7 2AZ
    Chem Commun (Camb) 47:400-2. 2011
    We report a general method based on wide-field fluorescence imaging of single molecule photobleaching and the Chung-Kennedy algorithm to measure the stoichiometry of functional protein complexes in living bacterial cells.
  37. ncbi Encoding microcarriers by spatial selective photobleaching
    Kevin Braeckmans
    Lab General Biochemistry and Physical Pharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium
    Nat Mater 2:169-73. 2003
    ..Patterns can be written in fluorescently dyed microspheres by 'spatial selective photobleaching' and can be identified by confocal microscopy...
  38. ncbi Loss of image quality in photobleaching during microscopic imaging of fluorescent probes bound to chromatin
    Tytus Bernas
    University of Silesia, Faculty of Biology and Protection of Environment, Department of Plant Anatomy and Cytology, Jagiellonska 28, Katowice, Poland
    J Biomed Opt 10:064015. 2005
    ..We apply the three measures to study the effect of photobleaching in cell nuclei stained with propidium iodide (PI) and chromomycin A3 (CA3) and imaged with fluorescence ..
  39. doi Measuring dynamics of nuclear proteins by photobleaching
    Miroslav Dundr
    National Cancer Institute, Bethesda, Maryland, USA
    Curr Protoc Cell Biol . 2003
    The mobility of nuclear proteins can be studied by photobleaching techniques...
  40. ncbi Acceptor-photobleaching FRET analysis of core kinetochore and NAC proteins in living human cells
    D Hellwig
    Department of Molecular Biology, Leibniz Institute for Age Research, Fritz Lipmann Institute, Jena, Germany
    Eur Biophys J 38:781-91. 2009
    ..and nucleosome-associated complex (NAC) proteins were analysed in living human interphase cells by acceptor photobleaching FRET...
  41. ncbi Irreversible photobleaching of bacteriorhodopsin in a high-temperature intermediate state
    Yasunori Yokoyama
    Department of Biotechnology, Tokyo University of Agriculture and Technology, Nakacho, Koganei, Tokyo 184 8588, Japan
    J Biochem 131:785-90. 2002
    ..As the recovery of the photocycle to the ground state and irreversible photobleaching to the denatured state may occur from the same local energy minimum, depending on the temperature, the ..
  42. ncbi The relationship between protoporphyrin IX photobleaching during real-time dermatological methyl-aminolevulinate photodynamic therapy (MAL-PDT) and subsequent clinical outcome
    Jessica S Tyrrell
    Clinical Photobiology, Peninsula Medical School, University of Exeter, Knowledge Spa, Royal Cornwall Hospital, Treliske, Truro, Cornwall TR1 3HD, UK
    Lasers Surg Med 42:613-9. 2010
    The relationship between protoporphyrin IX (PpIX) photobleaching and cellular damage during aminolevulinic (ALA) photodynamic therapy (PDT) has been studied at the cellular level...
  43. ncbi Enhanced macromolecular diffusion in brain extracellular space in mouse models of vasogenic edema measured by cortical surface photobleaching
    Marios C Papadopoulos
    Department of Medicine and Physiology, University of California, San Francisco, California 94143 0521, USA
    FASEB J 19:425-7. 2005
    ..of an inert macromolecular fluorescent marker (FITC-dextran, 70 kDa) in the ECS by fluorescence recovery after photobleaching after staining the exposed cerebral cortex in vivo...
  44. ncbi Analysis of photobleaching in single-molecule multicolor excitation and Förster resonance energy transfer measurements
    Christian Eggeling
    Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Gottingen, Germany
    J Phys Chem A 110:2979-95. 2006
    Dye photobleaching is a major constraint of fluorescence readout within a range of applications...
  45. ncbi Fluorescence recovery after photobleaching on the confocal laser-scanning microscope: generalized model without restriction on the size of the photobleached disk
    Nick Smisdom
    Hasselt University, Transnational University Limburg, Biomedical Research Institute, School of Life Sciences, Agoralaan Building C, Diepenbeek 3590, Belgium
    J Biomed Opt 16:046021. 2011
    Fluorescence recovery after photobleaching (FRAP) carried out on a confocal laser-scanning microscope (CLSM) performs well for photobleached disks that are large compared to the resolution of the bleaching beam...
  46. ncbi Single molecule blinking and photobleaching separated by wide-field fluorescence microscopy
    Thomas Gensch
    Institute for Biological Information Processing, IBI 1, Research Centre Jülich GmbH, 52425 Julich, Germany
    J Phys Chem A 109:6652-8. 2005
    ..Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density...
  47. ncbi The use of fluorescence redistribution after photobleaching for analysis of cellular microtubule dynamics
    Claire E Walczak
    Medical Sciences, Indiana University, Bloomington, Indiana 47405, USA
    Methods Cell Biol 97:35-52. 2010
    ..present approaches to assess MT dynamics in cultured mammalian cells using fluorescence redistribution after photobleaching. We include a discussion of cell culture and imaging conditions that maintain cell viability...
  48. pmc Accelerated photobleaching of a cyanine dye in the presence of a ternary target DNA, PNA probe, dye catalytic complex: a molecular diagnostic
    M Wang
    Investigen Inc, Hercules, California, USA
    Anal Chem 81:2043-52. 2009
    ..Here we report a novel, rapid nucleic acid detection method based on the accelerated photobleaching of the light-sensitive cyanine dye, 3,3'-diethylthiacarbocyanine iodide (DiSC(2)(3) I(-)), in the presence of ..
  49. pmc Protoporphyrin IX fluorescence photobleaching increases with the use of fractionated irradiation in the esophagus
    Brian W Pogue
    Dartmouth College, Thayer School of Engineering, Hanover, New Hampshire 03755, USA
    J Biomed Opt 13:034009. 2008
    ..Reducing the optical irradiance from 50 to 25 mWcm did not significantly alter photobleaching in rat esophagus, but fractionation of the light at 1-min on and off intervals did increase photobleaching up ..
  50. ncbi Signal amplification between Gbetagamma release and PI3Kgamma-mediated PI(3,4,5)P3 formation monitored by a fluorescent Gbetagamma biosensor protein and repetitive two component total internal reflection/fluorescence redistribution after photobleaching an
    Astrid Tannert
    Department of Molecular Pharmacology and Cell Biology, Neurowissenschaftliches Forschungszentrum, Charite Universitatsmedizin Berlin, Berlin, Germany
    Biochemistry 47:11239-50. 2008
    ..Here, changes in mobility were monitored by fluorescence redistribution after photobleaching (FRAP) which was repetitively applied before and after stimulation of cells...
  51. ncbi Photobleaching-based dosimetry predicts deposited dose in ALA-PpIX PDT of rodent esophagus
    Chao Sheng
    Thayer School of Engineering, Dartmouth College, Harvard Medical School, Boston, MA, USA
    Photochem Photobiol 83:738-48. 2007
    ..The underlying assumption in the interpretation of the data is that rapid photobleaching of PpIX occurs when there is ample oxygen supply, and this bleaching is not present when oxygen is limited...
  52. ncbi Diffusion of alpha-chymotrypsin in solution-crowded media. A fluorescence recovery after photobleaching study
    Isabel Pastor
    Department of Physical Chemistry and Research Institute of Theoretical and Computational Chemistry of University of Barcelona, Barcelona, Spain
    J Phys Chem B 114:4028-34. 2010
    Fluorescence recovery after photobleaching (FRAP) is one of the most powerful and used techniques to study diffusion processes of macromolecules in membranes or in bulk...
  53. pmc Pulsed interleaved excitation
    Barbara K Müller
    Physical Chemistry, Department of Chemistry and Biochemistry, and Center for NanoScience, Ludwig Maximilians Universitat Munchen, Butenandstrasse 11 Haus E, D 81377 Munich, Germany
    Biophys J 89:3508-22. 2005
    ..We also show the application of PIE to spFRET measurements, where complexes that have low FRET efficiency can be distinguished from those that do not have an active acceptor...
  54. pmc Quantifying green fluorescent protein diffusion in Escherichia coli by using continuous photobleaching with evanescent illumination
    Kristin M Slade
    Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599 3290, USA
    J Phys Chem B 113:4837-45. 2009
    Fluorescence recovery after photobleaching and fluorescence correlation spectroscopy are the primary means for studying translational diffusion in biological systems...
  55. pmc Continuous photobleaching in vesicles and living cells: a measure of diffusion and compartmentation
    A Delon
    Laboratoire de Spectrometrie Physique, Centre National de la Recherche Scientifique, UMR5588, Universite Joseph Fourier, Saint Martin d Heres, France
    Biophys J 90:2548-62. 2006
    We present a comprehensive and analytical treatment of continuous photobleaching in a compartment, under single photon excitation. In the very short time regime (t<0.1 ms), the diffusion does not play any role...
  56. pmc The CRAC channel consists of a tetramer formed by Stim-induced dimerization of Orai dimers
    Aubin Penna
    Department of Physiology and Biophysics, University of California Irvine, California 92697 4561, USA
    Nature 456:116-20. 2008
    ..of green fluorescent protein (GFP)-tagged Orai expressed in Xenopus oocytes showed predominantly two-step photobleaching, again consistent with a dimeric basal state...
  57. ncbi Photobleaching of retinal pigment epithelium melanosomes reduces their ability to inhibit iron-induced peroxidation of lipids
    Andrzej Zadlo
    Department of Biophysics, Jagiellonian University, Krakow, Poland
    Pigment Cell Res 20:52-60. 2007
    ..We have previously shown that a similar loss in the content of the RPE melanin occurs during human lifetime, which may suggest that the normal antioxidant properties of human RPE melanin become compromised with aging...
  58. ncbi Photobleaching on photonic crystal enhanced fluorescence surfaces
    Vikram Chaudhery
    Department of Electrical and Computer Engineering, University of Illinois at Urbana Champaign, 1406 W Green St, Urbana, IL 61801, USA
    J Fluoresc 21:707-14. 2011
    The effect of resonant fluorescent enhancement from a photonic crystal surface upon the fluorescent photobleaching rate of Cyanine-5 labeled protein has been investigated...
  59. pmc A quantitative approach to analyze binding diffusion kinetics by confocal FRAP
    Minchul Kang
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA
    Biophys J 99:2737-47. 2010
    ..diffusion of both the unbound and bound species, and also compensates for binding diffusion that occurs during photobleaching, a critical consideration in confocal FRAP analysis...
  60. ncbi Molecular photobleaching kinetics of Rhodamine 6G by one- and two-photon induced confocal fluorescence microscopy
    Christian Eggeling
    Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Gottingen, Germany
    Chemphyschem 6:791-804. 2005
    ..Herein, we report on the dependence of photobleaching on the excitation conditions, using the dye Rhodamine 6G as a typical example...
  61. pmc Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes
    Hari Shroff
    Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA
    Proc Natl Acad Sci U S A 104:20308-13. 2007
    ..The simplicity, minimal invasiveness, resolution, and speed of the technique all suggest its potential to directly visualize molecular interactions within cellular structures at the nanometer scale...
  62. pmc Monitoring photobleaching and hemodynamic responses to HPPH-mediated photodynamic therapy of head and neck cancer: a case report
    Ulas Sunar
    Department of Cell Stress Biology, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA
    Opt Express 18:14969-78. 2010
    ..We quantified blood flow, oxygenation and HPPH drug photobleaching before and after therapeutic light treatment by utilizing fast, non-invasive diffuse optical methods...
  63. pmc Poleward transport of Eg5 by dynein-dynactin in Xenopus laevis egg extract spindles
    Marianne Uteng
    Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany
    J Cell Biol 182:715-26. 2008
    ..How motors move and interact inside spindles is unknown. Using photoactivation and photobleaching, we measure mitotic motor movement inside a dynamic spindle...
  64. ncbi Photobleaching approaches to investigate diffusional mobility and trafficking of Ras in living cells
    J Shawn Goodwin
    Department of Molecular Physiology and Biophysics, 718 Light Hall, Vanderbilt University School of Medicine, Nashville, TN 37232, USA
    Methods 37:154-64. 2005
    ..We focus on three techniques, fluorescence recovery after photobleaching (FRAP), fluorescence loss in photobleaching (FLIP), and selective photobleaching...
  65. ncbi Intersection of ChIP and FLIP, genomic methods to study the dynamics of the cohesin proteins
    Adrian J McNairn
    Stowers Institute for Medical Research, 1000 E 50th Street, Kansas City, MO 64110, USA
    Chromosome Res 17:155-63. 2009
    ..We present evidence from fluorescence loss in photobleaching (FLIP) experiments in budding yeast that the decay constant of each cohesin subunit is approximately 60-90 s ..
  66. ncbi Selective visualization of fluorescent sterols in Caenorhabditis elegans by bleach-rate-based image segmentation
    Daniel Wüstner
    Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK 5230 Odense M, Denmark
    Traffic 11:440-54. 2010
    ..Our approach is generally useful for identifying fluorescent probes in the presence of high cellular autofluorescence...
  67. ncbi Loss of melanin from human RPE with aging: possible role of melanin photooxidation
    Tadeusz Sarna
    Department of Biophysics, Jan Zurzycki Institute of Molecular Biology and Biotechnology, Jagiellonian University, ul Gronostajowa 7, 30 387 Krakow, Poland
    Exp Eye Res 76:89-98. 2003
    ..Irradiation of RPE in eyecups with intense visible light was found to produce a time-dependent photobleaching of melanosomes that was accompanied by the formation of hydrogen peroxide...
  68. ncbi Repair machinery of symbiotic photosynthesis as the primary target of heat stress for reef-building corals
    Shunichi Takahashi
    Faculty of Science, University of the Ryukyus, Okinawa, 903 0213 Japan
    Plant Cell Physiol 45:251-5. 2004
    ..Acropora digitifera was exposed to light, a moderate increase of temperature induced coral bleaching through photobleaching of algal pigments, but not through expulsion of symbiotic algae...
  69. ncbi Dynamics of proteins in Golgi membranes: comparisons between mammalian and plant cells highlighted by photobleaching techniques
    T H Ward
    Immunology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom
    Cell Mol Life Sci 61:172-85. 2004
    ..In particular, we focus on the technique of selective photobleaching known as fluorescence recovery after photobleaching, which has successfully shed light on essential ..
  70. pmc Photobleaching-corrected FRET efficiency imaging of live cells
    Tomasz Zal
    Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA
    Biophys J 86:3923-39. 2004
    ..This makes it difficult to evaluate and compare FRET imaging data between laboratories. Furthermore, photobleaching poses a problem for FRET imaging in timelapse experiments and three-dimensional reconstructions...
  71. ncbi Photobleaching, mobility, and compartmentalisation: inferences in fluorescence correlation spectroscopy
    A Delon
    LSP, CNRS UMR5588, Universite Joseph Fourier, BP 87, 38402 Saint Martin d Heres, France
    J Fluoresc 14:255-67. 2004
    ..estimated experimentally from Fluorescence Correlation Spectroscopy (FCS) combined with the measurement of the photobleaching rate...
  72. ncbi Patterning protein concentration using laser-assisted adsorption by photobleaching, LAPAP
    Jonathan M Belisle
    Maisonneuve Rosemont Hospital, University of Montreal, QC, Canada
    Lab Chip 8:2164-7. 2008
    ..The method exploits photobleaching of fluorescently tagged molecules to generate patterns and concentration gradients of protein with sub-micron ..
  73. ncbi Effect of cholesterol on lateral diffusion of fluorescent lipid probes in native hippocampal membranes
    Thomas J Pucadyil
    Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India
    Chem Phys Lipids 143:11-21. 2006
    ..structure, in native and cholesterol-depleted hippocampal membranes using the fluorescence recovery after photobleaching (FRAP) approach...
  74. pmc Monolayers of silver nanoparticles decrease photobleaching: application to muscle myofibrils
    P Muthu
    Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, Texas 76107 2699, USA
    Biophys J 95:3429-38. 2008
    ..This causes photodamage of labeled proteins and rapid photobleaching of the fluorophores...
  75. ncbi Minimizing photobleaching during confocal microscopy of fluorescent probes bound to chromatin: role of anoxia and photon flux
    T Bernas
    Laboratory of Confocal Microscopy and Image Analysis, Department of Biophysics, Faculty of Biotechnology, Jagiellonian University, Gronostajowa 7, 30 387 Krakow, Poland
    J Microsc 215:281-96. 2004
    Exposure to light can destroy the ability of a molecule to fluoresce. Such photobleaching limits the use of fluorescence and confocal microscopy in biological studies...
  76. ncbi Photobleaching reveals complex effects of inhibitors on transcribing RNA polymerase II in living cells
    Maud Fromaget
    Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, UK
    Exp Cell Res 313:3026-33. 2007
    RNA polymerase II transcribes most eukaryotic genes. Photobleaching studies have revealed that living Chinese hamster ovary cells expressing the catalytic subunit of the polymerase tagged with the green fluorescent protein contain a large ..
  77. ncbi Fluorescence photobleaching analysis for the study of cellular dynamics
    Nectarios Klonis
    Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia
    Eur Biophys J 31:36-51. 2002
    ..and the emergence of green fluorescent protein (GFP) transfection technology has led to the increasing use of photobleaching studies to examine aspects of cellular dynamics in living cells...
  78. ncbi Stoichiometry and turnover in single, functioning membrane protein complexes
    Mark C Leake
    Clarendon Laboratory, Department of Physics, University of Oxford, Parks Road, Oxford OX1 3PU, UK
    Nature 443:355-8. 2006
    ..Counting fluorophores by the stepwise photobleaching of single GFP molecules showed that each motor contains approximately 22 copies of GFP-MotB, consistent with ..
  79. pmc Expanding two-photon intravital microscopy to the infrared by means of optical parametric oscillator
    Josephine Herz
    Cecilie Vogt Klinik, Charite University Medicine Berlin, Berlin, Germany
    Biophys J 98:715-23. 2010
    ..and 2), higher penetration depths in tissue (up to 80%) at higher resolution and with reduced photobleaching and phototoxicity...
  80. ncbi Photobleaching and photoactivation: following protein dynamics in living cells
    Jennifer Lippincott-Schwartz
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Cell Biol . 2003
    ..Here, we discuss the techniques of photobleaching and photoactivation, which can reveal the location and movement of proteins...
  81. pmc Effects of organelle shape on fluorescence recovery after photobleaching
    Ivo F Sbalzarini
    Institute of Computational Science, and Institute of Biochemistry, ETH Zurich, 8092 Zurich, Switzerland
    Biophys J 89:1482-92. 2005
    ..Fluorescence recovery after photobleaching (FRAP) experiments are performed in tissue culture cells expressing GFP-KDEL, a soluble, fluorescent protein, ..
  82. pmc Irradiance-dependent photobleaching and pain in delta-aminolevulinic acid-photodynamic therapy of superficial basal cell carcinomas
    William J Cottrell
    Department of Imaging Sciences, University of Rochester, Rochester, New York, USA
    Clin Cancer Res 14:4475-83. 2008
    ..High irradiance and/or photosensitizer levels cause inefficient treatment from oxygen depletion in preclinical models...
  83. pmc Circumvention of fluorophore photobleaching in fluorescence fluctuation experiments: a beam scanning approach
    Dmitri Satsoura
    Department of Biochemistry and Biomedical Sciences, McMaster University, 1200 Main Street West, Hamilton, ON, L8N3Z5, Canada
    Chemphyschem 8:834-48. 2007
    b>Photobleaching is a fluorophore-damaging process that commonly afflicts single-molecule fluorescence studies. It becomes an especially severe problem in fluorescence fluctuation experiments when studying slowly diffusing particles...
  84. ncbi Photobleaching in two-photon scanning fluorescence correlation spectroscopy
    Zdeněk Petrášek
    Biophysics Group, Biotechnologisches Zentrum, Technische Universitat Dresden, Tatzberg 47 51, 01307 Dresden, Germany
    Chemphyschem 9:147-58. 2008
    ..correlation spectroscopy (FCS) is often preferred to one-photon excitation because of reduced bulk photobleaching and photodamage, and deeper penetration into scattering media, such as thick biological specimens...
  85. ncbi Nonblinking and long-lasting single-molecule fluorescence imaging
    Ivan Rasnik
    Department of Physics, Emory University, Atlanta, Georgia 30322, USA
    Nat Methods 3:891-3. 2006
    b>Photobleaching and blinking of fluorophores pose fundamental limitations on the information content of single-molecule fluorescence measurements...
  86. ncbi Suppression of photobleaching-induced artifacts in frequency-domain FLIM by permutation of the recording order
    Erik B van Munster
    Molecular Cytology Section, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands
    Cytometry A 58:185-94. 2004
    b>Photobleaching can lead to significant errors in frequency-domain fluorescence lifetime imaging microscopy (FLIM)...
  87. ncbi Concatenation of cyan and yellow fluorescent proteins for efficient resonance energy transfer
    Satoshi Shimozono
    Laboratory for Cell Function Dynamics, Brain Science Institute, The Institute of Physical and Chemical Research RIKEN, 2 1 Hirosawa, Wako City, Saitama 351 0198, Japan
    Biochemistry 45:6267-71. 2006
    ..Moreover, Cy11.5 was used as an optical highlighter after photobleaching of YFP. When HeLa cells expressing Cy11.5 were irradiated at 514...
  88. ncbi Photobleaching of YFP does not produce a CFP-like species that affects FRET measurements
    Sophie E Verrier
    Nat Methods 3:491-2; author reply 492-3. 2006
  89. pmc Visual cycle: Dependence of retinol production and removal on photoproduct decay and cell morphology
    Petri Ala-Laurila
    Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118, USA
    J Gen Physiol 128:153-69. 2006
    ..The more rapid rates of retinol production in cones compared to rods are consistent with the more rapid operation of the visual cycle in these cells...
  90. ncbi Photobleaching of YFP does not produce a CFP-like species that affects FRET measurements
    Christopher Thaler
    Nat Methods 3:491; author reply 492-3. 2006
  91. pmc An oxygen scavenging system for improvement of dye stability in single-molecule fluorescence experiments
    Colin Echeverría Aitken
    Biophysics Program, Stanford University School of Medicine, Stanford, California 94305 5126, USA
    Biophys J 94:1826-35. 2008
    ..We observed increased stability of Cy3 and Alexa488 in the presence of the antioxidants ascorbic acid and n-propyl gallate. This new O(2)-scavenging system should have wide application for single-molecule fluorescence experiments...
  92. ncbi YFP photoconversion revisited: confirmation of the CFP-like species
    Michael T Kirber
    Nat Methods 4:767-8. 2007
  93. ncbi An ultrastructural readout of fluorescence recovery after photobleaching using correlative light and electron microscopy
    Kevin J Darcy
    MRC Laboratory for Molecular Cell Biology and Cell Biology Unit, University College London, Gower Street, London, WC1E 6BT, UK
    Nat Protoc 1:988-94. 2006
    Fluorescence recovery after photobleaching (FRAP) provides an important quantitative readout of the mobility of fluorescently tagged structures in live tissue...
  94. ncbi Membrane protein stoichiometry determined from the step-wise photobleaching of dye-labelled subunits
    Somes K Das
    Department of Molecular and Cellular Medicine, The Texas A and M University System Health Science Center, College Station, TX 77843 1114, USA
    Chembiochem 8:994-9. 2007
  95. ncbi Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging
    R A Hoebe
    Center for Advanced Microscopy, Section of Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 316, 1098 SM Amsterdam, The Netherlands
    Nat Biotechnol 25:249-53. 2007
    ..However, fluorescence live-cell imaging is limited by photobleaching and phototoxicity induced by the excitation light...
  96. ncbi The M3/M4 cytoplasmic loop of the alpha1 subunit restricts GABAARs lateral mobility: a study using fluorescence recovery after photobleaching
    Macarena Peran
    Department of Anatomy and Human Embryology, Faculty of Medicine, University of Granada, Granada, Spain
    Cell Motil Cytoskeleton 63:747-57. 2006
    ..Previous studies using Fluorescence Recovery After Photobleaching (FRAP) have shown a restricted mobility in GABAARs containing the alpha1 subunit...
  97. doi Measuring protein mobility by photobleaching GFP chimeras in living cells
    Erik L Snapp
    NICHD, National Institutes of Health, Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland, USA
    Curr Protoc Cell Biol . 2003
    This unit describes fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) using commercially available confocal scanning laser microscopy (CSLM)...
  98. ncbi Fluorescence resonance energy transfer (FRET) measurement by gradual acceptor photobleaching
    E B Van Munster
    Centre for Advanced Microscopy, Section Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, The Netherlands
    J Microsc 218:253-62. 2005
    ..One of the techniques for measuring FRET is acceptor photobleaching: the increase in donor fluorescence after complete acceptor photobleaching is a measure of the FRET efficiency...
  99. ncbi Quantitative determination of the reduction of phototoxicity and photobleaching by controlled light exposure microscopy
    R A Hoebe
    Centre for Microscopy Research CMO, Department of Cell Biology and Histology, Academic Medical Centre, Amsterdam, The Netherlands
    J Microsc 231:9-20. 2008
    Phototoxicity and photobleaching are major limitations in live-cell fluorescence microscopy. They are caused by fluorophores in an excited singlet or triplet state that generate singlet oxygen and other reactive oxygen species...
  100. ncbi Rapid actin transport during cell protrusion
    Daniel Zicha
    Light Microscopy, Cancer Research UK, Lincoln s Inn Fields Laboratories, London WC2A 3PX, UK
    Science 300:142-5. 2003
    ..The recently developed FLAP (fluorescence localization after photobleaching) method permits the tracking of one fluorophore after localized photobleaching by using the other as a ..
  101. ncbi Fluorescence resonance energy transfer of GFP and YFP by spectral imaging and quantitative acceptor photobleaching
    C Dinant
    Department of Pathology, Josephine Nefkens Institute, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands
    J Microsc 231:97-104. 2008
    ..We adapted the linear unmixing procedure from commercially available software (Zeiss) for use with acceptor photobleaching FRET using GFP and YFP as FRET pair. FRET efficiencies up to 52% for a GFP-YFP fusion protein were measured...

Research Grants71

  1. Function and Expression of Connexins in the pre-Botzinger Complex
    JONATHAN KELTY; Fiscal Year: 2007
    ..or CBX) and a control agent (glycyrrhizic acid or GZA), will be evaluated by a fluorescence recovery after photobleaching (FRAP) microscopy technique...
  2. Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex
    Derek Toomre; Fiscal Year: 2009
    ..Consistent with the recovery act's goal, this funding will provide full-time summer employment for 4 undergraduate students and accelerate scientific achievement of the parent DP2 award...
  3. STRUCTURE AND FUNCTION OF PAXILLIN
    Christopher E Turner; Fiscal Year: 2012
    ..fibroblasts and utilize confocal fluorescence time-lapse microscopy, combined with Fluorescence Recovery after Photobleaching (FRAP) and Fluorescence Resonance Energy Transfer (FRET) analysis to evaluate cell morphology, polarity and ..
  4. The Role of Connexin32 in the Pathogensis of CMTX
    STEVEN SIMON SCHERER; Fiscal Year: 2013
    ..functional GJ plaques will be investigated - by immunostaining, scrape loading, fluorescence recovery after photobleaching (FRAP), and electrophysiology...
  5. Role of Cofilin and Wnt Signaling in Neural Crest Development
    O apos NEIL WIGGAN; Fiscal Year: 2013
    ..Dynamic F-actin remodeling will be assessed by techniques such as fluorescence recovery after photobleaching. Furthermore, by use of commonly utilized biochemical procedures we will establish the means by which Wnt ..
  6. Dynamics of Transcriptional Regulation
    Elena Zelin; Fiscal Year: 2011
    ..The fluorescence recovery after photobleaching (FRAP) technique will be used in conjunction with engineered cell lines that contain hsp70 loci arrays...
  7. Histone proline isomerization and gene regulation
    Nobuaki Kikyo; Fiscal Year: 2010
    ..pluripotency in embryonic stem cells will be studied by genetic knockdown, DNA microarray and the fluorescence photobleaching technique...
  8. Physiology of Class III PI 3-kinase Signaling 2
    Jonathan M Backer; Fiscal Year: 2013
    ..the dynamics of hVps34 complex formation, both at the whole cell level and, using fluorescence recovery after photobleaching (FRAP), on the autophagosomal membrane...
  9. Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging Course
    TERRI I GRODZICKER; Fiscal Year: 2013
    ..quantify, and analyze proteins in living cells using standard fluorescence, fluorescence recovery after photobleaching, and resonance energy transfer...
  10. Nikon TiE Motorized Microscope System For 3D Image Acquisition, Deconvolution, Li
    Robert S Fuller; Fiscal Year: 2010
    ..resolution for 3D image acquisition, deconvolution, and live cell time lapse and fluorescence recovery after photobleaching (FRAP)...
  11. Leica TCS SP5 Multiphoton Confocal Microscope
    Heather A Drummond; Fiscal Year: 2012
    ..of new imaging technologies, specifically multiphoton imaging technology which reduces tissue damage and photobleaching and offers imaging deeper into tissues...
  12. Real-time microviscosity measurement tools for the cell
    Mark A Haidekker; Fiscal Year: 2010
    ..viscosity on the microscopic scale is determined by fluorescence anisotropy, fluorescence recovery after photobleaching (FRAP), or magnetic nanoparticles...
  13. Genesis of Epithelial Cell Lumens
    Jennifer L Harder; Fiscal Year: 2013
    ..mutagenesis, 3-dimensional cyst assay, custom cell surface trafficking assay, and fluorescence recovery after photobleaching. In Aim #2, she will evaluate an as yet unexplored mechanism of controlling polarity, that cells use ..
  14. THE ROLES OF ESPINS IN HAIR CELL STEREOCILIA
    James R Bartles; Fiscal Year: 2012
    ..will use in-vitro binding assays, electron microscopy, small-angle x-ray scattering and confocal fluorescence-photobleaching assays to determine how espins with deafness mutations and wild-type espin isoforms differ in their actin-..
  15. Spinning Disk Confocoal Microscope
    Judy Drazba; Fiscal Year: 2010
    ..the ability to do fluorescence resonance energy transfer (FRET) measurements, fluorescence recovery after photobleaching (FRAP) measurements and photoactivation, thereby substantially advancing their NIH-funded biomedical research...
  16. A lipid-based approach towards a high-resolution structure of a functional nAChR
    Jose Lasalde-Dominicci; Fiscal Year: 2013
    ..assess the lipid composition nAChR-DCs, and it is innovative in its use of the fluorescent recovery after photobleaching (FRAP) approach in the LCP to estimate nAChR mobile fraction and diffusion coefficient to correlate with ..
  17. Optical studies of the cone photoreceptor synapse
    Richard H Kramer; Fiscal Year: 2013
    ..markers of synaptic vesicles to measure vesicle mobility on the ribbon with Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS)...
  18. Molecular Mechanisms of PTH-Mediated Trafficking in Renal Tubular Cells
    Judith T Blaine; Fiscal Year: 2013
    ..cells will be examined using total internal reflection fluorescence microscopy and fluorescence recovery after photobleaching. These imaging techniques will be complemented by investigation of the effects of PTH on the cellular ..
  19. Satiety signaling in Caenorhabditis elegans
    Leon Avery; Fiscal Year: 2013
    ..Develop methods to measure synaptic activity by recovery of fluorescence after photobleaching of the synaptic vesicle pH sensor synaptopHluorin...
  20. Internal Dynamics of the Postsynaptic Density
    Thomas A Blanpied; Fiscal Year: 2013
    ..fluorescence morphometry approach for examining single, living PSDs, and have developed two high-resolution photobleaching assays for monitoring the movement of proteins within them...
  21. The Role of Small Ankyrin 1 in the Sarcoplasmic Reticulum in Skeletal Muscle
    Andrew P Ziman; Fiscal Year: 2011
    ..brevis muscle of the rat, including assays of the continuity of the stores by fluorescence recovery after photobleaching (FRAP), and Ca2+ release by activators of the ryanodine receptor and by electrical stimulation;and (2) to ..
  22. Fluorophore-Conjugated Antibodies for Imaging and Resection of GI Tumors
    Meng Yang; Fiscal Year: 2013
    ..2 We will compare several different fluorophores for in vivo dosing response, in vivo signal duration, in vivo photobleaching, and in vivo signal-to-background ratio in our mouse models of human pancreatic cancer...
  23. Analysis of Z-band assembly and maintenance in living skeletal muscle cells
    JOSEPH WILLIAM SANGER; Fiscal Year: 2013
    ..FRAP (Fluorescence Recovery After Photobleaching) experiments will measure the changing dynamics of key proteins in the z-bodies and Z-bands during ..
  24. SIGNAL TRANSDUCTION AT FERTILIZATION
    Laurinda A Jaffe; Fiscal Year: 2012
    ..oocytes with fluorescent tracers, and analysis by 2-photon microscopy and fluorescence recovery after photobleaching. These studies will be complemented by investigations of the phosphorylation of connexin 43 on specific ..
  25. An Animal Model for Cornelia de Lange Syndrome
    DALE L DORSETT; Fiscal Year: 2012
    ..in cohesion factors affect cohesin chromosome-binding dynamics in vivo using fluorescence recovery after photobleaching. It is hoped that insights from these studies will shed light on the mechanisms by which small changes in ..
  26. Control of Vascular Cell Motility by CaMKII
    Harold A Singer; Fiscal Year: 2013
    ..Fluorescence recovery after photobleaching (FRAP) analysis of GFP-tagged focal adhesion components will be used to assess CaMKII4-dependent regulation of ..
  27. Biophysics Cell Membrane Probe
    William E Brownell; Fiscal Year: 2010
    ..is modular so that a variety of user specific experiments such as calcium imaging, fluorescence recovery after photobleaching or fluorescence resonance energy transfer may be easily accommodated...
  28. CRCNS: Receptor mobility and scaffold dynamics at single glutamatergic synapses
    Sridhar Raghavachari; Fiscal Year: 2013
    ..account for receptor mobility, we will use high-throughput single-molecule tracking PALM and high-resolution photobleaching and photoactivation in synaptic subdomains to measure protein mobility within the synapse...
  29. MECHANICAL LOADING AND BONE
    Hiroki Yokota; Fiscal Year: 2013
    ..electronic speckle pattern interferometry as well as molecular transport using fluorescence recovery after photobleaching;(3) conducting bone histomorphometry to evaluate ex vivo data;and (4) examining load-driven adverse effects ..
  30. ZEISS LSM710 SCANNING CONFOCAL MICROSCOPE
    Hazel L Sive; Fiscal Year: 2010
    ..The LSM710 can perform FRAP (Fluorescence Recovery After Photobleaching) to analyze molecular dynamics;calcium imaging is simple and targeted uncaging of fluorescent indicators or ..
  31. Reticulon Function in ER Morphology
    Gia Voeltz; Fiscal Year: 2013
    ..For these studies, we will be using fluorescence microscopy, immunoprecipitations, fluorescence recovery after photobleaching, and electron microscopy with immunogold labeling...
  32. Creating Safe Biodegradable Photoluminescent Implant Polymers
    Jian Yang; Fiscal Year: 2013
    ..As alternatives, fluorescent dyes suffer from photobleaching and fluorescent proteins are dim in vivo and hard to manipulate...
  33. In vivo and crude extract analysis of polyQ aggregation intermediates
    Robert Fairman; Fiscal Year: 2012
    ..This technique will be used, combined with FRAP (fluorescence recovery after photobleaching, carried out using a confocal microscope), to probe the aggregation states of polyQ-containing proteins both ..
  34. Assembly and Axonal Transport of Neurofilaments
    Anthony Brown; Fiscal Year: 2013
    ..We will use cell fusion as well as photobleaching and photoactivation strategies to test these hypotheses in cultured neurons...
  35. Intravital Multiphoton Microscope
    Florea Lupu; Fiscal Year: 2010
    ..Two- and 3-photon imaging will minimize photo-damage and photobleaching, maximize penetration into thick specimens, permit excitation of UV-absorbing fluorophores with red and near ..
  36. Regulation of ENaC by phosphoinositides, MARCKS, and the cytoskeleton
    ABDEL AYUBE ALLI; Fiscal Year: 2012
    ..We will perform Fluorescence Recovery after Photobleaching (FRAP) to determine the role of the actin cytoskeleton in the formation of a membrane signaling complex of ..
  37. Solute Transport in the Bone Lacunar-Canalicular System
    Liyun Wang; Fiscal Year: 2012
    ..To this end, we recently developed a new imaging method based on Fluorescence Recovery After Photobleaching (FRAP) that allows measurement of solute movement in the bone LCS in situ and in real-time (Wang et al.,2005...
  38. Biophysical Basis of Organization and Dynamics of Prestin Membrane Complexes
    Robert M Raphael; Fiscal Year: 2013
    ..resonance energy transfer (FRET), fluorescence lifetime imaging (FLIM), fluorescence recovery after photobleaching (FRAP) and single molecule imaging...
  39. University of Minnesota Multi-User Total Internal Reflection Fluorescence Microsc
    Lorene M Lanier; Fiscal Year: 2010
    ..In addition, the sample is exposed to a considerable amount of fluorescent light, which can cause photobleaching (which leads to loss of imaging capacity) and, in the case of live cell imaging, phototoxicity and cell death...
  40. Sensitive and multiplexed analysis of cancer biomarkers with QD barcodes
    Xiaohu Gao; Fiscal Year: 2012
    ..g., tunable light emission, improved signal brightness, resistance against photobleaching, and simultaneous excitation of multiple colors) and our ability to make optical barcodes using these ..
  41. Olympus BX-DSU Live-Cell Disk Scanning Imaging / Electrophysiology System
    Bradley K Taylor; Fiscal Year: 2010
    ..of laser confocal microscopes is the very intense excitation light, leading to substantial dye decomposition (photobleaching) during live-cell imaging...
  42. STRUCTURE/FUNCTION OF SHORT WAVELENGTH VISUAL PIGMENTS
    Barry E Knox; Fiscal Year: 2011
    ..how the cone opsin family has attained absorption maxima from 350 nm to 650 nm while achieving the rapid photobleaching and regeneration properties that play a large role in shaping the light responses of cones...
  43. Acquisition of a Carl Zeiss Cell Observer Spinning Disk Confocal Microscope
    J Michael McCaffery; Fiscal Year: 2013
    ..acquisition speeds;and the ability to accurately modulate laser power to greatly minimize phototoxicity and photobleaching during extended time-lapse imaging sessions...
  44. Mechanism of Targeting of a Luminal KDEL-containing Protein to the Golgi Complex
    DEBORAH ARONSON; Fiscal Year: 2010
    ..Live cell fluorescence microscopy photobleaching and biochemistry techniques will be employed to distinguish whether GC localization occurs via retention or ..
  45. Light Activation of Retinal Insulin Receptor Signaling
    RAJU VS RAJALA; Fiscal Year: 2013
    ..novel pathway uses growth factor receptor bound protein (Grb14), an upstream regulator of IR, and requires photobleaching of rhodopsin for membrane targeting...
  46. Effects of NHERF1 expression on PTH1R dynamics and function in polarized cells
    DAVID SHERWOOD WHEELER; Fiscal Year: 2012
    ..These studies will be conducted using various live-cell imaging techniques such as fluorescence recover after photobleaching, quantum dot single molecule tracking, optical measurements of cAMP and calcium and image cross-correlation ..
  47. MOLECULAR MECHANISM OF PHOTORECEPTOR G PROTEIN SIGNALING
    Nikolai O Artemyev; Fiscal Year: 2013
    ..The mutant Gat1 models will be examined with EGFP imaging, immunofluorescence, and Fluorescence Recovery After Photobleaching (FRAP) analysis of lateral and longitudinal diffusion...
  48. Regulation of activity-dependent ProSAp2 synaptic dynamics
    MAGALI HOLT ROWAN; Fiscal Year: 2011
    ..Live-cell time-lapse confocal microscopy and Fluorescence Recovery After Photobleaching (FRAP) will be used here to assess the dynamic exchange properties of ProSAP2 in response to pharmacological ..
  49. Analysis of the interplay between cell adhesion and bacterial protein translocati
    Ralph R Isberg; Fiscal Year: 2013
    ..In addition, a combined photoactivation/photobleaching strategy will be introduced to the microbial pathogenesis field, in order to identify the site in the cell ..
  50. Determining the mechanisms that regulate neurofascin155 distribution in myelin
    Jeffrey L Dupree; Fiscal Year: 2010
    ..Following transfection of these constructs, I will employ Fluorescent Recovery After Photobleaching to quantify the role that palmitoylaiton plays in the regulation of NF155 cluster maintenance...
  51. Columbia AFM System Equipment Grant
    Clark T Hung; Fiscal Year: 2010
    ..provides photoactivation or bleaching and simultaneous image collection, enabling fluorescence recovery after photobleaching (FRAP) and molecular uncaging applications...
  52. The Coordination Chemistry of Nanocrystal Surfaces: Understanding the Role of Val
    BRANDI MICHELLE COSSAIRT; Fiscal Year: 2011
    ..Their extraordinary extinction coefficients, fluorescence quantum yields, and stability toward photobleaching have lead many researchers to investigate nanocrystals as fluorescent probes in biology...
  53. Photobiology of Rhodopsin and the Cone Pigments
    ROBERT RICHARDS BIRGE; Fiscal Year: 2012
    ..have absorption maxima that range from 350 to 660 nm, and upon the absorption of light, undergo an efficient photobleaching sequence to produce an activated protein...
  54. Compact all-fiber based CARS microscope for biomedical imaging
    LEYUN ZHU; Fiscal Year: 2012
    ..Furthermore, the fluorophores themselves are subject to photobleaching which results in limited imaging time and prevents long term studies...
  55. P2X channel mobility in the plasma membrane of neurons
    Esther Richler; Fiscal Year: 2011
    ..The specific goals of this proposal are 1) to use fluorescence recovery after photobleaching (FRAP) and single particle tracking (SPT) of Quantum dot labelled P2X2 receptees to probe the cellular ..
  56. Shared TIRF and Patterned Illumination Fluorescence Microscope
    Aaron F Straight; Fiscal Year: 2010
    ..of live cell time-lapse fluorescence and brightfield microscopy, TIRF microscopy, patterned fluorescence photobleaching and photoactivation and iterative deconvolution...
  57. Automated Pre-screening System for in meso Membrane protein crystallization
    VADIM GENNADYEVICH CHEREZOV; Fiscal Year: 2011
    ..We propose to develop an automated system for carrying out Fluorescence Recovery after Photobleaching (FRAP) studies of membrane proteins in meso...
  58. Automated Pre-screening System for in meso Membrane protein crystallization
    VADIM GENNADYEVICH CHEREZOV; Fiscal Year: 2012
    ..We propose to develop an automated system for carrying out Fluorescence Recovery after Photobleaching (FRAP) studies of membrane proteins in meso...
  59. Expression and Characterization of alpha6beta2beta3 nicotinic receptors
    BRANDON JARROD HENDERSON; Fiscal Year: 2013
    ..Combining fluorescent nAChR subunits and photobleaching techniques will allow the quantification of specific subunits in receptor assemblies...
  60. Dye-Based biosensors: simultaneous imaging of multiple protein activities
    KLAUS MICHAEL HAHN; Fiscal Year: 2011
    ..We have studied the mechanisms of dye photobleaching and response to solvent polarity, and devised novel approaches to enhance water solubility...
  61. Next-generation Fluorescent Probes for Biological Research
    Scott C Blanchard; Fiscal Year: 2013
    ..Such phenomena, which include both transient (blinking) and irreversible (photobleaching), add uncertainties to all fluorescence applications, and are particularly limiting for single-molecule ..
  62. INTRACELLULAR TRANSPORT OF AMPHIPATHS
    Bruce Luxon; Fiscal Year: 2000
    ..bile acid analog dansyl taurocholate, will be measured by two dimensional fluorescence recovery after photobleaching (FRAP)...
  63. Tumor cell and microenvironment changes causing antiangiogenic therapy resistance
    Manish Aghi; Fiscal Year: 2013
    ..vivo models of anti-angiogenic therapy resistance and an innovative application of fluorescence recovery after photobleaching (FRAP) to correlate integrin mobility and turnover in focal adhesions with drug resistance...
  64. ERYTHROCYTE ADHESION IN FALCIPARUM MALARIA
    IRWIN SHERMAN; Fiscal Year: 2004
    ..with electron micrographic immunolocalization of adhesive sites, as well as by the use of fluorescence photobleaching recovery, polarized fluorescence depletion and luminescence quenching...
  65. MITOSIS IN NORMAL AND NEOPLASTIC CELLS
    BILL BRINKLEY; Fiscal Year: 2006
    ..relationships of centromeric chromatin and associated proteins using live cell imaging and quantitative photobleaching, (3) define and characterize the molecular determinants (genetic/epigenetic) that specify the site for ..
  66. Novel Epigenetic Marks in Mitosis
    KENNETH S contact ZARET; Fiscal Year: 2010
    ..Our recent mechanistic studies using fluorescence recovery after photobleaching (FRAP) show that FoxA and PARP binding to chromatin is far more stable than that of most other transcription ..
  67. LASER PHOTORADIATION THERAPY OF MALIGNANT TUMORS
    Michael Berns; Fiscal Year: 2002
    ..kinetics (2) subcellular localization/binding (3) subcellular phototoxicity (4) photosensitizer photobleaching and predictive value of fluorescence for phototoxicity (5) mechanism(s) of cellular uptake...
  68. Deltavision RT Image Restoration System for Established Research Core Facility
    Thomas Keller; Fiscal Year: 2007
    ..for cells susceptible to photodamage or where the fluorescent molecules to be imaged are susceptible to photobleaching or dim, in experiments in which two or more colors of fluorescently labeled molecules are interacting, for ..
  69. ACAS 570 ANCHORED CELL ANALYZER AND SORTER
    RAYMOND HESTER; Fiscal Year: 1991
    ..additional study without subjecting the cells to detachment protocols, (4) fluorescence redistribution after photobleaching to analyze inter- and intracellular translocation of membrane components, cytoplasmic structures and ..
  70. BIOMEDICAL RESEARCH SUPPORT
    SAMUEL SHACKS; Fiscal Year: 1991
    ..channel plate-coupled intensified CCD video camera for previewing, selecting and focussing fields with minimal photobleaching and photodamage; analytical imaging workstation including computer, software and data storage for 3-..
  71. Stoichiometry and Architecture of the Vesicle Fusion Machine in PC-12 Cells
    JAMES WEISSHAAR; Fiscal Year: 2009
    ..PALM uses a long sequence of activation-location-photobleaching cycles to locate all labeled copies of a protein one by one...