I am working on plant cell culture and I need to find out the percentage of viable cells in the cell suspension using UV-spectophotometer, apart from staining and counting cells using heamocytometer. Please suggest a method.
The followig literature may be useful for your research
The results of this study highlight the importance of taking into account the time-point at which cells are observed and whether the cells are light-grown and chloroplast-containing or not, for any study on plant AL-PCD(Apoptotic-Like), as it appears that chloroplasts can play a significant role in AL-PCD regulation
Chloroplast and reactive oxygen species involvement in apoptotic-like programmed cell death in Arabidopsis suspension cultures; by Siamsa M. Doyle, Mark Diamond and Paul F. McCabe*
Journal of Exoerimental Botany; 61(2), October 12, 2009,473-482
Dear Deepthi,
Add Evan’s Blue to a final concentration of 200 microliter/ml and mix by gently inverting the closed tube three times. Remove the supernatant after 4 min incubation at room temperature (depending on your cells this can extend to even 10 min - do some preliminary experiments to optimise), when the cells had settled to the bottom. Wash cells twice with culture medium, centrifuging at low speed in a bench-top centrifuge and removing the supernatant after each wash. Then use a haemocytometer or even mount on a slide and count total cells to those that are stained blue. Blue stained cells are dead. You don't need a UV spec for this. Another method is FDA staining if you have a fluorescence microscope. For these studies, you should ideally have a well dispersed culture because it is hard to get the stain to middle of cell clumps and counting is also difficult.
Good luck!
@ Ranjith Pathirana Thank you sir for your kind reply. It was really very helpful.
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