Should you collect dead floating cells as well and harvesting live cells when performing the Trypan blue assay?
The TRYPAN BLUE ASSAY will enable you to accurately determine the cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the hemacytometer. If cells take up trypan blue, they are considered non-viable.
If you are using the trypan blue assay for viability experiments, then yes, otherwise you are overestimating the proportion of viable cells. If you are just subculturing your cells, this isn't necessary, you mainly want to know how many living cells you have to calculate your desired cell density.
Hi Dahia, if you are determining the viability of adherent cells, then no.
Floating cells are dead and will not give you any information about the viability of the adherent cells. You are only going to subculture live cells so you need to know their number. You will know that the cells are dying off if they don't grow.
Adrian gave the correct information.
If you need to know how many viable cells you have (for example, after some treatment), then you have to include also detached/non adherent cells. Conversely, if you just need to subculture your cells, you have to exclude non adherent/detached cells from your counting procedure, because you have to consider alive cells only when you calculate the amount of cell suspension for the subsequent cell seeding.
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