Dear Flow-ers A couple of points for discussion. What is the consensus as to "wash" or "not-to-wash" human blood cells after Annexin-V FITC staining. Most protocols do not wash. If you wash in what buffer, Annexin Ca++ binding buffer, PBS etc.? This subject has been touched on briefly in the past but not resolved. Secondly, if staining for apoptotic cells with PI after ethanol fixation and subsequent analysis for a sub-G1 peak, do investigators apply the same strict criteria as to CVs of G0/G1 peaks as if they were solely performing cell cycle analysis, i.e. <3% for a diploid human lymphocyte population? Len Brown Flow Cytometry Mater Medical Research Institute Brisbane, Australia
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