Check cell viablity after FACS - (Apr/23/2012 )

Hi all,

I have some GFP positive cells that I sort by FACS. However, they don't look very happy when I put them in culture after the sorting and I think most of them are not surviving.

Could anyone recomend me any live/dead cell assays to check the viability after the sorting? Can I do this by adding PI during the sorting?

Many thanks in advance for the information!

Take an aliquot and add some trypan blue...live dead count on haemocytometer.

You can add PI to selectively stain dead cells and even gate them out when you analyze your FACS data - have a look here. As bob1 was saying, you can also count live (unstained) and dead (stained) cells with a hemocytometer and get the viability from there.