Message text written by INTERNET:David.McFarland@mcmail.vanderbilt.edu >Hello everyone. I have a client that wants to do analysis of RBCs. This is new to me. Seems everyone else wants to make sure we avoid the RBCs! Basically, they are injecting mice with a FITC-labeled ligand that should only be bound (and internalized?) by cells that express the receptor for said ligand. They have control mice, and transgenics that express the receptor. The question is where is it expressed? We did some earlier experiments excluding the RBCs, but now they want to look at them as well. They are looking to me for advice and I don't have any to give them. So, does anyone have some good advice to pass along? I'll even give you the credit! David McFarland< We often study human RBC (and platelets) as well as leucocytes. RBC are predominantly slightly smaller than lymphocytes and of low granularity (for locating in FSC/SSC profiles) but this is heterogeneous and varies. RBC are 100 - 1000 fold more numerous than leucocytes, so will predominate in any whole blood sample - so scale your sample size accordingly. Obviously - don't lyse. Since there will be size/granularity overlap with leucocytes use another marker to discriminate - such as glycophorin A for RBC or CD45 for leucocytes. I don't know what is equivalent for these in mice - but I hope this gives some ideas. Robin Barclay SNBTS Edinburgh
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