CHO Fixation - PFA %'s and other such (Jan/13/2009 )

Hello all

Does anyone have advice on fixation techniques for a CHO cell line. I've tried .1% gluteraldehyde with 2% PFA with a full time course, and I can't get a good fixation...I thought the time course would tease out the answer, but maybe a PFA % issue?

What will you use the fixed cells for? Different fixation techniques are used for different processes. I typically use ice cold 2% PFA in 1x PBS for 10 min on ice for fixing cells on a slide/coverslip. If you just want to look at structures, you could try ice cold 1:1 Acetone:ethanol for 10 min on ice, though this shrinks/dehydrates the cells a bit.