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Int. J. Mol. Sci. 2010, 11(3), 956-966; doi:10.3390/ijms11030956
Article

Light Dose is a Limiting Factor to Maintain Cell Viability in Fluorescence Microscopy and Single Molecule Detection

1
,
1
,
1
,
2
,
2
 and
1,2,*
1 Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany 2 Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany
* Author to whom correspondence should be addressed.
Received: 14 January 2010 / Revised: 20 February 2010 / Accepted: 21 February 2010 / Published: 8 March 2010
(This article belongs to the Special Issue Single Molecules)
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Abstract

A test system for cell viability based on colony formation has been established and applied to high resolution fluorescence microscopy and single molecule detection. Living cells were irradiated either by epi-illumination or by total internal reflection (TIR) of a laser beam, and light doses where at least 90% of irradiated cells survived were determined. These light doses were in the range of a few J/cm2 up to about 200 J/cm2 depending on the wavelength of illumination as well as on the presence or absence of a fluorescent dye (e.g., the membrane marker laurdan). In general, cells were less sensitive to TIR than to epi-illumination. However, comparably high light doses needed for repetitive excitation of single molecules limit the application of super-resolution microscopy to living cells.
Keywords: cell viability; light dose; fluorescence microscopy; TIR; single molecules cell viability; light dose; fluorescence microscopy; TIR; single molecules
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Wagner, M.; Weber, P.; Bruns, T.; Strauss, W.S.L.; Wittig, R.; Schneckenburger, H. Light Dose is a Limiting Factor to Maintain Cell Viability in Fluorescence Microscopy and Single Molecule Detection. Int. J. Mol. Sci. 2010, 11, 956-966.

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