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Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging

(2007) NATURE BIOTECHNOLOGY. 25(2). p.249-253
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Abstract
Fluorescence microscopy of living cells enables visualization of the dynamics and interactions of intracellular molecules. However, fluorescence live-cell imaging is limited by photobleaching and phototoxicity induced by the excitation light. Here we describe controlled light-exposure microscopy ( CLEM), a simple imaging approach that reduces photobleaching and phototoxicity two- to tenfold, depending on the fluorophore distribution in the object. By spatially controlling the light-exposure time, CLEM reduces the excitation-light dose without compromising image quality. We show that CLEM reduces photobleaching sevenfold in tobacco plant cells expressing microtubule-associated GFP-MAP4 and reduces production of reactive oxygen species eightfold and prolongs cell survival sixfold in HeLa cells expressing chromatin-associated H2B-GFP. In addition, CLEM increases the dynamic range of the fluorescence intensity at least twofold.
Keywords
LIVING CELLS, CONFOCAL MICROSCOPY, PHOTODYNAMIC THERAPY, 2-STEP PHOTOLYSIS, KINETICS, PHOTON, MITOSIS, STRESS, PLANTS, STATE

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Chicago
Hoebe, RA, CH Van Oven, TWJ Gadella Jr, Pankaj Dhonukshe, CJF Van Noorden, and EMM Manders. 2007. “Controlled Light-exposure Microscopy Reduces Photobleaching and Phototoxicity in Fluorescence Live-cell Imaging.” Nature Biotechnology 25 (2): 249–253.
APA
Hoebe, RA, Van Oven, C., Gadella Jr, T., Dhonukshe, P., Van Noorden, C., & Manders, E. (2007). Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging. NATURE BIOTECHNOLOGY, 25(2), 249–253.
Vancouver
1.
Hoebe R, Van Oven C, Gadella Jr T, Dhonukshe P, Van Noorden C, Manders E. Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging. NATURE BIOTECHNOLOGY. 2007;25(2):249–53.
MLA
Hoebe, RA, CH Van Oven, TWJ Gadella Jr, et al. “Controlled Light-exposure Microscopy Reduces Photobleaching and Phototoxicity in Fluorescence Live-cell Imaging.” NATURE BIOTECHNOLOGY 25.2 (2007): 249–253. Print.
@article{3202861,
  abstract     = {Fluorescence microscopy of living cells enables visualization of the dynamics and interactions of intracellular molecules. However, fluorescence live-cell imaging is limited by photobleaching and phototoxicity induced by the excitation light. Here we describe controlled light-exposure microscopy ( CLEM), a simple imaging approach that reduces photobleaching and phototoxicity two- to tenfold, depending on the fluorophore distribution in the object. By spatially controlling the light-exposure time, CLEM reduces the excitation-light dose without compromising image quality. We show that CLEM reduces photobleaching sevenfold in tobacco plant cells expressing microtubule-associated GFP-MAP4 and reduces production of reactive oxygen species eightfold and prolongs cell survival sixfold in HeLa cells expressing chromatin-associated H2B-GFP. In addition, CLEM increases the dynamic range of the fluorescence intensity at least twofold.},
  author       = {Hoebe, RA and Van Oven, CH and Gadella Jr, TWJ and Dhonukshe, Pankaj and Van Noorden, CJF and Manders, EMM},
  issn         = {1087-0156},
  journal      = {NATURE BIOTECHNOLOGY},
  language     = {eng},
  number       = {2},
  pages        = {249--253},
  title        = {Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging},
  url          = {http://dx.doi.org/10.1038/nbt1278},
  volume       = {25},
  year         = {2007},
}

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