Light dose is a limiting factor to maintain cell viability in fluorescence microscopy and single molecule detection.
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.
Wagner
Michael
M
Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
Weber
Petra
P
Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
Bruns
Thomas
T
Institut für Angewandte Forschung, Hochschule Aalen, Beethovenstr. 1, D-73430 Aalen, Germany.
Strauss
Wolfgang S L
WSL
Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany.
Wittig
Rainer
R
Institut für Lasertechnologien in der Medizin und Meßtechnik an der Universität Ulm, Helmholtzstr. 12, D-89081 Ulm, Germany.
Schneckenburger
Herbert
H
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.
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*