How long are cells viable in PBS? - (to make trypan blue exclusion assay less tedious) (Jul/23/2009 )

Hi,

I am going to have to do a lot of cell counts soon which I find incredibly boring. Once I've lifted the cells with trypsin, spun down and resuspended them in PBS, how long will they be viable for before staining with trypan blue?

I guess what I'm asking is can I do the cell counts in batches i.e. count for half an hour, go off and do something else for a bit then come back and do more counts, or do I have to sit there for a couple of hours and do them all in one go?

Thanks,
P

Penguin on Jul 23 2009, 08:41 AM said:

For a consistency you need to do them at one go. But if intersample consistency is not that important, you can do them in batches.

Every minute that the cells are left in PBS, a certain number of them die. It depends upon what cells you are using that how fast a significant numbers of them go dead.

In any case, do the following.

1. Keep the cells on ice.

1. Resuspend them in culture media, rather than the PBS.

This two measures will make the cells survive longer.

cellcounter on Jul 23 2009, 09:08 PM said:

Yeah, thats what I was thinking, I guess I'll just have to get on with it!! It won't be so bad if I have the radio on!
Thanks,
P

The best thing as suggested by the other person is to keep cells in media. You can re-suspend the cells in plain media and keep for a longer time. But keep them in ice otherwise it wont work well.

Also, it will help you in better cell counting due to lesser cell death and debris thereof.