In need of advice....currently working on tryphan blue assay...just wonder that in the protocol mention that upon 24 hour treatment, the cells need to be trypsinized, centrifuged and the pelleted cells is stained with tryphan blue.Non-viable cells will be blue, viable cells will be unstained.My question here is during centrifuge most dead cell will be left in supernatant in which will be discarded as only pellet cells will be used. Therefore is this assay reliable?
May be this research paper will help:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075294/
Yes it is reliable assay since only dead cells with damaged cell membrane absorb the dye and become colored
Hi, Yes I agree with Rodolphe Hajj.
It is reliable. but It is important whtat speed you will use. I use that recommended for my cell line e.g. 700xg.
Have a nice assay ;)
Regards
Thank you all for your informative suggestion. I will definitely follow your advice. Thank you
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