title: Light dose is a limiting factor to maintain cell viability in fluorescence microscopy and single molecule detection.
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/cm(2) up to about 200 J/cm(2) 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.
authors.affiliation: Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Wagner
authors.affiliation: Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Weber
authors.affiliation: Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bruns
authors.affiliation: Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany.
authors.forename: Wolfgang S L
authors.initials: WSL
authors.lastname: Strauss
authors.affiliation: Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Wittig
authors.affiliation: Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.|Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany.
authors.forename: Herbert
authors.initials: H
authors.lastname: Schneckenburger
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000528:radiation effects; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D011829:Radiation Dosage; D014466:Ultraviolet Rays / Q000009:adverse effects*