Hi Abdul Rouf,

Usually to get viable cell count serial dilution method is used. Only those will grow which are living so you can pickup colonies from the petri plate on which you have grown. e.g take your sample and spread it on the selective media petri plate then identify and pick up colonies. ( in case of bacteria)

Hi Samreen Ela ,Thank you for your answer.Moreover,I want to know the separation process for non- adherent cancer cell lines.For example;If we cultivate the non-adherent cancer cell line in a flask and after 24- 48 hours, we get to observe the dead cells along-with live cells in suspension and live cells are suspended in such a way that they have formed some clumps with dead cells and debris which seems to be very important to separate them for further purpose.How can we do that?

Hi Abdul Rouf in my opinion maybe you can use centrifugation process. As you have 3 constituents in your sample so they must have the different densities and will settle down differently.

You can use a vital die like trypan blue to identify dead cells (which will stain blue). A weak trypsin (or other protease) solution will dissociate debris from live cells, especially with mild agitation, like trituration with a wide bore pipet. Slow centrifugation will then enrich the pellet with live cells, leaving smaller debris in suspension.

Wash with 1xpbs gently and remove the pbs. Add fresh media. If it is difficult to grow then add some hfcs