FACs Analysis
http://metadb.riken.jp/db/IMPC_RDF/Impress_procedure/ESLIM_018_001
The characterisation of of main leukocyte subsets in peripheral blood cells from mice by flow cytometry.Reliable values of frequencies of leukocyte clusters are very much dependent on the appropriate preparation, acquisition and gating of leukocytes. This methodinvolves ammonium chloride erythrocyte lyses, thus preventing interference by larger amounts of erythrocytes. For all stainings, a monoclonal antibody to thecell surface glycoprotein CD45 is used to be able to create a CD45+ gate, allowing discrimination of leukocytes from debris, erythrocytes and thrombocytes. Furthermore, staining with propidium iodide (PI) gates out dead cells. Samples are acquired until a number of 30000 living CD45+ cells is reached for each sample (ref Adler and Busch 2007, submitted). Data are compensated analysed and using FlowJo software.This protocol is part of the EMPReSSslim pipeline and is carried out on a cohort of 10 males and 10 females (age matched) at a time.