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. Tannert A, Voigt P, Burgold S, Tannert S, Schaefer M. 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 photobleachi. Biochemistry. 2008;47:11239-50 pubmed publisher
    ..Here, changes in mobility were monitored by fluorescence redistribution after photobleaching (FRAP) which was repetitively applied before and after stimulation of cells...
  2. Belisle J, Correia J, Wiseman P, Kennedy T, Costantino S. Patterning protein concentration using laser-assisted adsorption by photobleaching, LAPAP. Lab Chip. 2008;8:2164-7 pubmed publisher
    ..The method exploits photobleaching of fluorescently tagged molecules to generate patterns and concentration gradients of protein with sub-micron ..
  3. Jarvi M, Patterson M, Wilson B. Insights into photodynamic therapy dosimetry: simultaneous singlet oxygen luminescence and photosensitizer photobleaching measurements. Biophys J. 2012;102:661-71 pubmed publisher
    ..The results show that SOL was always predictive of cytotoxicity and immune to PDT's complex dynamics, whereas photobleaching-based dosimetry failed under hypoxic conditions...
  4. Uteng M, Hentrich C, Miura K, Bieling P, Surrey T. Poleward transport of Eg5 by dynein-dynactin in Xenopus laevis egg extract spindles. J Cell Biol. 2008;182:715-26 pubmed publisher
    ..How motors move and interact inside spindles is unknown. Using photoactivation and photobleaching, we measure mitotic motor movement inside a dynamic spindle...
  5. Cottrell W, Paquette A, Keymel K, Foster T, Oseroff A. Irradiance-dependent photobleaching and pain in delta-aminolevulinic acid-photodynamic therapy of superficial basal cell carcinomas. Clin Cancer Res. 2008;14:4475-83 pubmed publisher
    ..Self-sensitized photobleaching of protoporphyrin IX (PpIX) fluorescence was used as a surrogate metric for photodynamic dose...
  6. McNairn A, Gerton J. Intersection of ChIP and FLIP, genomic methods to study the dynamics of the cohesin proteins. Chromosome Res. 2009;17:155-63 pubmed publisher
    ..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 ..
  7. Heinze K, Costantino S, De Koninck P, Wiseman P. Beyond photobleaching, laser illumination unbinds fluorescent proteins. J Phys Chem B. 2009;113:5225-33 pubmed publisher
    ..Beyond the common problems of photobleaching and phototoxicity, we present evidence that photounbinding also has the potential to compromise such methods, ..
  8. Khan P, Barik A, Vinod E, Sangunni K, Jain H, Adarsh K. Coexistence of fast photodarkening and slow photobleaching in Ge19As21Se60 thin films. Opt Express. 2012;20:12416-21 pubmed publisher
    ..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...
  9. Buch C, Hunt M, Alexson S, Hallberg E. Localization of peroxisomal matrix proteins by photobleaching. Biochem Biophys Res Commun. 2009;388:355-9 pubmed publisher
    ..We have used photobleaching in live cells expressing green fluorescent protein (GFP)-fusion proteins to show that imported bona fide ..

More Information

Publications62

  1. Arumugam S, Lee T, Benkovic S. Investigation of stoichiometry of T4 bacteriophage helicase loader protein (gp59). J Biol Chem. 2009;284:29283-9 pubmed publisher
    ..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 ..
  2. Shi X, Lim J, Ha T. Acidification of the oxygen scavenging system in single-molecule fluorescence studies: in situ sensing with a ratiometric dual-emission probe. Anal Chem. 2010;82:6132-8 pubmed publisher
    ..are labeled with small organic dyes which suffer from their limited photostability evidenced by blinking and photobleaching. An enzymatic oxygen scavenging system of glucose oxidase and catalase is widely used to improve the dye ..
  3. Lenn T, Gkekas C, Bernard L, Engl C, Jovanovic G, Buck M, et al. Measuring the stoichiometry of functional PspA complexes in living bacterial cells by single molecule photobleaching. Chem Commun (Camb). 2011;47:400-2 pubmed publisher
    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.
  4. Sunar U, Rohrbach D, Rigual N, Tracy E, Keymel K, Cooper M, et al. Monitoring photobleaching and hemodynamic responses to HPPH-mediated photodynamic therapy of head and neck cancer: a case report. Opt Express. 2010;18:14969-78 pubmed publisher
    ..We quantified blood flow, oxygenation and HPPH drug photobleaching before and after therapeutic light treatment by utilizing fast, non-invasive diffuse optical methods...
  5. Chaudhery V, Lu M, Huang C, George S, Cunningham B. Photobleaching on photonic crystal enhanced fluorescence surfaces. J Fluoresc. 2011;21:707-14 pubmed publisher
    The effect of resonant fluorescent enhancement from a photonic crystal surface upon the fluorescent photobleaching rate of Cyanine-5 labeled protein has been investigated...
  6. Wang M, Holmes Davis R, Rafinski Z, Jedrzejewska B, Choi K, Zwick M, et al. Accelerated photobleaching of a cyanine dye in the presence of a ternary target DNA, PNA probe, dye catalytic complex: a molecular diagnostic. Anal Chem. 2009;81:2043-52 pubmed publisher
    ..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 ..
  7. Wang K, Cottrell W, Mitra S, Oseroff A, Foster T. Simulations of measured photobleaching kinetics in human basal cell carcinomas suggest blood flow reductions during ALA-PDT. Lasers Surg Med. 2009;41:686-96 pubmed publisher
    ..Protoporphyrin IX (PpIX) photobleaching in the lesion and the adjacent perilesion normal margin was monitored by fluorescence spectroscopy...
  8. Wüstner D, Landt Larsen A, Faergeman N, Brewer J, Sage D. Selective visualization of fluorescent sterols in Caenorhabditis elegans by bleach-rate-based image segmentation. Traffic. 2010;11:440-54 pubmed publisher
    ..Our approach is generally useful for identifying fluorescent probes in the presence of high cellular autofluorescence. ..
  9. Douplik A, Chen D, Akens M, Zanati S, Cirocco M, Bassett N, et al. Assessment of photobleaching during endoscopic autofluorescence imaging of the lower GI tract. Lasers Surg Med. 2010;42:224-31 pubmed publisher
    ..The present study was designed to quantify the degree of fluorescence photobleaching induced by the excitation light during use in the colon, since significant photobleaching could lead to false ..
  10. Zhang Y, Liu X, Osburn C, Wang M, Qin B, Zhou Y. Photobleaching response of different sources of chromophoric dissolved organic matter exposed to natural solar radiation using absorption and excitation-emission matrix spectra. PLoS ONE. 2013;8:e77515 pubmed publisher
    ..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...
  11. Pogue B, Sheng C, Benevides J, Forcione D, Puricelli B, Nishioka N, et al. Protoporphyrin IX fluorescence photobleaching increases with the use of fractionated irradiation in the esophagus. J Biomed Opt. 2008;13:034009 pubmed publisher
    ..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 ..
  12. Muthu P, Calander N, Gryczynski I, Gryczynski Z, Talent J, Shtoyko T, et al. Monolayers of silver nanoparticles decrease photobleaching: application to muscle myofibrils. Biophys J. 2008;95:3429-38 pubmed publisher
    ..This causes photodamage of labeled proteins and rapid photobleaching of the fluorophores...
  13. Demuro A, Penna A, Safrina O, Yeromin A, Amcheslavsky A, Cahalan M, et al. Subunit stoichiometry of human Orai1 and Orai3 channels in closed and open states. Proc Natl Acad Sci U S A. 2011;108:17832-7 pubmed publisher
    We applied single-molecule photobleaching to investigate the stoichiometry of human Orai1 and Orai3 channels tagged with eGFP and expressed in mammalian cells...
  14. Altman R, Terry D, Zhou Z, Zheng Q, Geggier P, Kolster R, et al. Cyanine fluorophore derivatives with enhanced photostability. Nat Methods. 2011;9:68-71 pubmed publisher
    ..Such conjugation is a powerful means of improving the robustness of fluorescence-based applications demanding long-lived, nonblinking fluorescence emission. ..
  15. Wüstner D, Solanko L, Lund F, Sage D, Schroll H, Lomholt M. Quantitative fluorescence loss in photobleaching for analysis of protein transport and aggregation. BMC Bioinformatics. 2012;13:296 pubmed publisher
    Fluorescence loss in photobleaching (FLIP) is a widely used imaging technique, which provides information about protein dynamics in various cellular regions...
  16. Erdel F, Rippe K. Quantifying transient binding of ISWI chromatin remodelers in living cells by pixel-wise photobleaching profile evolution analysis. Proc Natl Acad Sci U S A. 2012;109:E3221-30 pubmed publisher
    ..To reach this goal, we have developed a fluorescence bleaching technique termed pixel-wise photobleaching profile evolution analysis (3PEA)...
  17. Hoebe R, Van der Voort H, Stap J, Van Noorden C, Manders E. Quantitative determination of the reduction of phototoxicity and photobleaching by controlled light exposure microscopy. J Microsc. 2008;231:9-20 pubmed publisher
    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...
  18. Duan C, Adam V, Byrdin M, Ridard J, Kieffer Jaquinod S, Morlot C, et al. Structural evidence for a two-regime photobleaching mechanism in a reversibly switchable fluorescent protein. J Am Chem Soc. 2013;135:15841-50 pubmed publisher
    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...
  19. Walczak C, Rizk R, Shaw S. The use of fluorescence redistribution after photobleaching for analysis of cellular microtubule dynamics. Methods Cell Biol. 2010;97:35-52 pubmed publisher
    ..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...
  20. Herz J, Siffrin V, Hauser A, Brandt A, Leuenberger T, Radbruch H, et al. Expanding two-photon intravital microscopy to the infrared by means of optical parametric oscillator. Biophys J. 2010;98:715-23 pubmed publisher
    ..and 2), higher penetration depths in tissue (up to 80%) at higher resolution and with reduced photobleaching and phototoxicity...
  21. Buzatu D, Cooper W, Summage West C, Sutherland J, Williams A, Bass D, et al. Photobleaching with phloxine B sensitizer to reduce food matrix interference for detection of Escherichia coli serotype O157:H7 in fresh spinach by flow cytometry. Food Microbiol. 2013;36:416-25 pubmed publisher
    ..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. ..
  22. Lemke E, Gambin Y, VanDelinder V, Brustad E, Liu H, Schultz P, et al. Microfluidic device for single-molecule experiments with enhanced photostability. J Am Chem Soc. 2009;131:13610-2 pubmed publisher
    ..b>Photobleaching is dramatically reduced by deoxygenation via gas diffusion through porous channel walls...
  23. Kang M, Day C, Dibenedetto E, Kenworthy A. A quantitative approach to analyze binding diffusion kinetics by confocal FRAP. Biophys J. 2010;99:2737-47 pubmed publisher
    ..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...
  24. Stasevich T, Mueller F, Michelman Ribeiro A, Rosales T, Knutson J, McNally J. Cross-validating FRAP and FCS to quantify the impact of photobleaching on in vivo binding estimates. Biophys J. 2010;99:3093-101 pubmed publisher
    ..To address this uncertainty, we compare fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) measurements of the binding kinetics of a transcription ..
  25. Gupta R, Srivastava O. Identification of interaction sites between human betaA3- and alphaA/alphaB-crystallins by mammalian two-hybrid and fluorescence resonance energy transfer acceptor photobleaching methods. J Biol Chem. 2009;284:18481-92 pubmed publisher
    ..The mammalian two-hybrid studies were complemented with fluorescence resonance energy transfer acceptor photobleaching studies using the above described mutant proteins, fused with DsRed (Red) and AcGFP fluorescent proteins...
  26. Dean K, Lubbeck J, Binder J, Schwall L, Jimenez R, Palmer A. Analysis of red-fluorescent proteins provides insight into dark-state conversion and photodegradation. Biophys J. 2011;101:961-9 pubmed publisher
    ..However, the brightness of FPs is limited by dark-state conversion (DSC) and irreversible photobleaching, which occur on different timescales...
  27. Smisdom N, Braeckmans K, Deschout H, VandeVen M, Rigo J, De Smedt S, et al. Fluorescence recovery after photobleaching on the confocal laser-scanning microscope: generalized model without restriction on the size of the photobleached disk. J Biomed Opt. 2011;16:046021 pubmed publisher
    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...
  28. Song S, Hanson M, Liu B, Chylack L, Liang J. Protein-protein interactions between lens vimentin and alphaB-crystallin using FRET acceptor photobleaching. Mol Vis. 2008;14:1282-7 pubmed
    ..FRET was estimated by the acceptor photobleaching method. Protein-protein interaction was evaluated by FRET efficiency...
  29. Penna A, Demuro A, Yeromin A, Zhang S, Safrina O, Parker I, et al. The CRAC channel consists of a tetramer formed by Stim-induced dimerization of Orai dimers. Nature. 2008;456:116-20 pubmed publisher
    ..of green fluorescent protein (GFP)-tagged Orai expressed in Xenopus oocytes showed predominantly two-step photobleaching, again consistent with a dimeric basal state...
  30. Dinant C, van Royen M, Vermeulen W, Houtsmuller A. Fluorescence resonance energy transfer of GFP and YFP by spectral imaging and quantitative acceptor photobleaching. J Microsc. 2008;231:97-104 pubmed publisher
    ..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...
  31. Munck S, Miskiewicz K, Sannerud R, Menchón S, Jose L, Heintzmann R, et al. Sub-diffraction imaging on standard microscopes through photobleaching microscopy with non-linear processing. J Cell Sci. 2012;125:2257-66 pubmed publisher
    ..However, such technology is still limited for many cell biologists. We present here a novel approach using photobleaching microscopy with non-linear processing (PiMP) for sub-diffraction imaging...
  32. Hines K. Inferring subunit stoichiometry from single molecule photobleaching. J Gen Physiol. 2013;141:737-46 pubmed publisher
    Single molecule photobleaching is a powerful tool for determining the stoichiometry of protein complexes...
  33. Tyrrell J, Campbell S, Curnow A. The relationship between protoporphyrin IX photobleaching during real-time dermatological methyl-aminolevulinate photodynamic therapy (MAL-PDT) and subsequent clinical outcome. Lasers Surg Med. 2010;42:613-9 pubmed publisher
    The relationship between protoporphyrin IX (PpIX) photobleaching and cellular damage during aminolevulinic (ALA) photodynamic therapy (PDT) has been studied at the cellular level...
  34. Ding H, Wong P, Lee E, Gafni A, Steel D. Determination of the oligomer size of amyloidogenic protein beta-amyloid(1-40) by single-molecule spectroscopy. Biophys J. 2009;97:912-21 pubmed publisher
    ..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, ..
  35. Adam V, Carpentier P, Violot S, Lelimousin M, Darnault C, Nienhaus G, et al. Structural basis of X-ray-induced transient photobleaching in a photoactivatable green fluorescent protein. J Am Chem Soc. 2009;131:18063-5 pubmed publisher
    ..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.
  36. Pastor I, Vilaseca E, Madurga S, Garcés J, Cascante M, Mas F. Diffusion of alpha-chymotrypsin in solution-crowded media. A fluorescence recovery after photobleaching study. J Phys Chem B. 2010;114:4028-34 pubmed publisher
    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...
  37. Onogi C, Hamaguchi H. Photobleaching of the "Raman spectroscopic signature of life" and mitochondrial activity in rho- budding yeast cells. J Phys Chem B. 2009;113:10942-5 pubmed publisher
    ..This signature showed photobleaching under irradiation of the 632.8 nm line of a He-Ne laser, only for the rho- strain...
  38. Hellwig D, Hoischen C, Ulbricht T, Diekmann S. Acceptor-photobleaching FRET analysis of core kinetochore and NAC proteins in living human cells. Eur Biophys J. 2009;38:781-91 pubmed publisher
    ..and nucleosome-associated complex (NAC) proteins were analysed in living human interphase cells by acceptor photobleaching FRET...
  39. Cordes T, Vogelsang J, Tinnefeld P. On the mechanism of Trolox as antiblinking and antibleaching reagent. J Am Chem Soc. 2009;131:5018-9 pubmed publisher
    Recent advances in photobleaching and blinking prevention have aided the advancement of single-molecule and super-resolution fluorescence microscopy...
  40. Slade K, Steele B, Pielak G, Thompson N. Quantifying green fluorescent protein diffusion in Escherichia coli by using continuous photobleaching with evanescent illumination. J Phys Chem B. 2009;113:4837-45 pubmed publisher
    Fluorescence recovery after photobleaching and fluorescence correlation spectroscopy are the primary means for studying translational diffusion in biological systems...
  41. Delon A, Usson Y, Derouard J, Biben T, Souchier C. Photobleaching, mobility, and compartmentalisation: inferences in fluorescence correlation spectroscopy. J Fluoresc. 2004;14:255-67 pubmed
    ..estimated experimentally from Fluorescence Correlation Spectroscopy (FCS) combined with the measurement of the photobleaching rate...
  42. Sheng C, Hoopes P, Hasan T, Pogue B. Photobleaching-based dosimetry predicts deposited dose in ALA-PpIX PDT of rodent esophagus. Photochem Photobiol. 2007;83:738-48 pubmed
    ..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...
  43. Braeckmans K, De Smedt S, Roelant C, Leblans M, Pauwels R, Demeester J. Encoding microcarriers by spatial selective photobleaching. Nat Mater. 2003;2:169-73 pubmed
    ..Patterns can be written in fluorescently dyed microspheres by 'spatial selective photobleaching' and can be identified by confocal microscopy...
  44. Ericson M, Grapengiesser S, Gudmundson F, Wennberg A, Larko O, Moan J, et al. A spectroscopic study of the photobleaching of protoporphyrin IX in solution. Lasers Med Sci. 2003;18:56-62 pubmed
    ..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...
  45. Zal T, Gascoigne N. Photobleaching-corrected FRET efficiency imaging of live cells. Biophys J. 2004;86:3923-39 pubmed
    ..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...
  46. Zadlo A, Rozanowska M, Burke J, Sarna T. Photobleaching of retinal pigment epithelium melanosomes reduces their ability to inhibit iron-induced peroxidation of lipids. Pigment Cell Res. 2007;20:52-60 pubmed
    ..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. ..
  47. Shroff H, Galbraith C, Galbraith J, White H, Gillette J, Olenych S, et al. Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes. Proc Natl Acad Sci U S A. 2007;104:20308-13 pubmed
    ..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. ..
  48. Dundr M, Misteli T. Measuring dynamics of nuclear proteins by photobleaching. Curr Protoc Cell Biol. 2003;Chapter 13:Unit 13.5 pubmed publisher
    The mobility of nuclear proteins can be studied by photobleaching techniques...
  49. Gensch T, Böhmer M, Aramendia P. Single molecule blinking and photobleaching separated by wide-field fluorescence microscopy. J Phys Chem A. 2005;109:6652-8 pubmed
    ..Atto590 shows very long blinking times and photobleaching and photoblinking that are both quadratically dependent on the irradiation power density...
  50. Delon A, Usson Y, Derouard J, Biben T, Souchier C. Continuous photobleaching in vesicles and living cells: a measure of diffusion and compartmentation. Biophys J. 2006;90:2548-62 pubmed
    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...
  51. Valentin G, Verheggen C, Piolot T, Neel H, Coppey Moisan M, Bertrand E. Photoconversion of YFP into a CFP-like species during acceptor photobleaching FRET experiments. Nat Methods. 2005;2:801 pubmed
  52. Petrášek Z, Schwille P. Photobleaching in two-photon scanning fluorescence correlation spectroscopy. Chemphyschem. 2008;9:147-58 pubmed
    ..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...
  53. Sarna T, Burke J, Korytowski W, Rózanowska M, Skumatz C, Zareba A, et al. Loss of melanin from human RPE with aging: possible role of melanin photooxidation. Exp Eye Res. 2003;76:89-98 pubmed
    ..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...