Fixing cells with PFA - (Jun/03/2010 )
I doing immunocytochemistry on cells from human blood samples. What I want to do is cytospin them onto a slide, fix them and then store the slide until I have accumulated a bunch of slides. Once I have a good number of slides, I will continue on with the rest of the experiment. Has anyone does this before? Is it possible? If so, how long have you stored the slides for? I was thinking store the slides in PBS at 4 degrees C.
PFA starts to lose its "fixing" after about a week at 4 deg C, such that the cells start to lose ultrastructure, which may or may not be important for you. If you want to store long term, fix in PFA, wash in PBS and then dehydrate in methanol/PBS starting with 20% MeOH and working up to 100% over a few steps. To use the slides after this, rehydrate by reversing the methanol/PBS steps.