| Autor: |
Hoebe RA; Center for Advanced Microscopy, Section of Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 316, 1098 SM Amsterdam, The Netherlands., Van Oven CH, Gadella TW Jr, Dhonukshe PB, Van Noorden CJ, Manders EM |
| Jazyk: |
angličtina |
| Zdroj: |
Nature biotechnology [Nat Biotechnol] 2007 Feb; Vol. 25 (2), pp. 249-53. Date of Electronic Publication: 2007 Jan 21. |
| DOI: |
10.1038/nbt1278 |
| Abstrakt: |
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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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