Cell Fixation for Long Storage Time

[Dionisia]

Hi,
Im working with mouse cells and i need a quick method to fix these cells and keep them for later analysis.
It will be mainly surface and intracellular antigens using flow cytometry.


Is there a good method for these types of samples and how long can we keep these samples before we label them?

What is also the best method for permeabilising these cells?
thanks

[ethidiumbromide]

When you say "cells", do you mean a tissue sample? or do you mean actual cells? And when you say "fix" what do you mean exactly? Freeze? Fix, as in fixing cells to a slide?

If cells, I think you should be able to freeze them -80 in cell culture media with 10% DMSO. If tissue, I'm not sure, but maybe just like it is?

When we fix our cells, we use 1-2% paraformaldehyde for 5-10, then wash 2-3 times with PBS. When we permeabilize cells we use 0.1% Triton X-100 in PBS for 2 minutes and then wash with PBS 2-3 times. But this protocol is for IF. I don't know if it would be different for flow.

If you can be more specific, I can ask someone in my lab on Monday and try to get you a protocol. We do flow all the time. Just not by me.