I'm actullay working on avian PBMCs,
and I've to use a fixative buffer because I can't do the acquisition of my flow cytometry assays at the end of the staining,
but when I used PBS + 1% formaldéhyde, I observed that my cells were killed or quite modified by this buffer,
Which other fixative buffer could i use in order to preserve cells and surface stainings ?
Have you tried methanol? Or one of the premade fixing buffers.
http://www.ncbi.nlm.nih.gov/pubmed/3543135
But generally speaking, wouldn't fixed cells always be dead?
I've always had good results with BD Cytofix. I've used it with PBMC and cell subsets from cattle, pigs, chickens and sheep.
It would help if you could explain how your cells were "modified". As Victoria said, they should be killed - that's the point of fixing them.
Hi Chapat
A cell fixative is used to fix cell proteins and it does kill cells. You could try other fixatives like methanol (in cold) - depends on what antigens you're looking to detect. Alternatively try fixing cells in PFA post staining the cells with the relevant antibodies.
Another better option is to fix the cells, wash off the fixative, store the cells in buffer overnight at 4oC, stain the next day and acquire on flow cytometer on the same day.
Best of luck!
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