fixation and permeabilization - (Sep/07/2014 )

Hi. 

I am new in flowcytometry.

I would like to detect intracellular antigen. My doubt is that, is it more appropriate to fix the cells prior to permeabilization then proceed to staining? Or should I permeabilize the cells without fixation? 

Usually fix then permeabilize.

Agree, this is the standard procedure. Unless of course you need the cells to survive the procedure like in cell sorting. Permeablization without fixing can apparently also be of use if the protein is close to the cell membrane or is a soluble protein in the cytoplasm, according to literature (but never tried this in particular)