I would like both monocytes, lymfocytes and granulocytes to eventually stain and stick on a slide. I used lymphoprep before, but this only gives me the mononuclear cells. I would like the complete leukocyte fraction. I think this could be done by simple centrifugation, but if anyone has any nice protocols that would be lovely.
Have you considered staining whole blood? You simply lyse the red blood cells before or after staining, and the other cell populations are unaffected.There should be published protocols out there (I have seen some on the BD Bioscience website) but I do not have one myself.
Zoya Georgieva
Hi Zoya, thanks for your answer. I would prefer to concentrate the leukocytes and then stain (since I need to quantify as well), so doing a smear is not ideal if that is what you mean. I suppose I could do erylysis and then just pelleting down the cells, but I am not a big fan of erylysis buffer. Although I think that will be the best option.
Thanks again!
Hi Jenny Bruijn,
Apologies, the technique I was suggesting works for flow cytometry, since I didn't notice you want to visualise the cells directly. I do not know if an equivalent protocol exists for immunocytochemistry.
This was the type of product and protocol I was suggesting (for use on a flow cytometer): https://www.bdbiosciences.com/ds/pm/tds/555899.pdf
I wonder if it is possible to stain as described in this, but prepare a slide from the resulting cell suspension at the end? I do not know if this would work (but it is probably what I would try if no other alternative is suggested!)
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