Adrian, A method that has been employed wih great success here at UK to help visualize bone marrow as well as PB abnormalities is by gating using a SSC-LOG (X) vs CD45 (Y) dot plot. By changing from linear to log and lowering your SSC PMT by about 100 you can see the familiar "paw print" of hematopoietic elements. As CD45 is employed (we use the Per-CP conjugated form from BD to allow 3 color analysis on a FACScan) the cells are separated nicely into quite distinct areas. As RBCs are CD45 negative they will fall to the bottom of the plot with mature lymphocytes at the top. Myelomonocytic elements will fall in between and be further separated by the SSC-Log amplification. Blasts will segregate below the lymphs and to the left of the monos and granulocyte precursors. With a little experience abnormalities will often be instantly obvious in their deviation from the norm. The beauty of this method really becomes apparent when an abnormality is less visually striking. Because of the nice separation it then becomes quite easy to obtain pure gates and regions for further analysis within a panel of MoAbs designed for lymphomas and leukemias. Barry Barry Grimes Manager, Hematopoiesis FACS Laboratory UKMC Hematopoiesis Center 800 Rose St. Lexington,KY 40536-0093 606-323-8193 bagrim1@pop.uky.edu
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