Imaging the division process in living tissue culture cells.
We detail some of the pitfalls encountered when following live cultured somatic cells by light microscopy during mitosis. Principle difficulties in this methodology arise from the necessity to compromise between maintaining the health of the cell while achieving the appropriate temporal and spatial resolutions required for the study. Although the quality of the data collected from fixed cells is restricted only by the quality of the imaging system and the optical properties of the specimen, the major limiting factor when viewing live cells is radiation damage induced during illumination. We discuss practical considerations for minimizing this damage, and for maintaining the general health of the cell, while it is being followed by multi-mode or multi-dimensional light microscopy.
Khodjakov
Alexey
A
Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509, USA. khodj@wadswoirth.org
Rieder
Conly L
CL
D000818:Animals; D018929:Cell Culture Techniques; D002455:Cell Division / Q000528:radiation effects*; D002478:Cells, Cultured; D007089:Image Enhancement; D007909:Lenses; D008027:Light; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D018715:Microscopy, Video / Q000295:instrumentation / Q000379:methods*; D013696:Temperature