Can anyone advise on lysis buffers which (i) will and (ii) will not permeabilise (for, say, PI exclusion/access), and similarly on fixation which would or would not permeabilise - and any combinations of lysis/fixation to have these opposite effects. Should you fix before lysing or lyse before fixing. I am interested in quantitative recovery and phenotypic (surface) assessment of rather small amounts of leucocytes from a lot of red cells. Does anyone have a good method for cell concentration or washing with minimal cell loss? There seems to be many ways of approaching this, so I would be glad to have some suggestions to weigh against my own initial ideas. Robin Barclay SNBTS: Edinburgh
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