Hello,
I am currently using MRC-5 cell lines and they are at 17th passage. I tried to collect and count them via trypan blue cell counting but I could not see enough cells even though they looked fine in the microscope. I used almost 16 T-25 flasks but couldn't catch enough cells. What could be the problem?
The Trypan Blue (TB) assay was proposed about a century ago and is still the most widely used method to perform cell viability analysis. So,
Trypan Blue has a greater affinity for serum proteins than for cellular protein. If the background is too dark, cells should be pelleted and resuspended in protein-free medium or salt solution prior to counting.
The trypan blue solution contained precipitate is still good to use. It can be warmed to dissolve the precipitate or it can be filtered to remove the precipitate.
good luck
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