Cell counting - (Jun/12/2005 )

Hi !

Recently I tested some chemicals to 293T cell and I need to count the cell number to know the effect of the chemicals for the cell growth.

But when I collect the cells to be a cell suspension and then will use tyrpan blue to stain the cells for counting, the cells aggregate as one by one. ( I use PBS-EDTA to split the cell from plates)

It let me not correctly count the cell number. I don't know how to solve this problem. Can someone give me some idea?

Any help very greatly appreciated !!

Hi,
I have always used trypsin EDTA to split the cells. After the cells come off (observe under an inverted m'scope) add serum-free media or just plain old 1XPBS. Dont do the entire batch at the same time if you have a lot of wells. Do them batch by batch just before countign.
Good luck
p..

If you need to use EDTA for some reason (as I do for my experiments as I cannot use trypsin) try using some collagenase as well to help dissassociate the cells. It works nicely.