Paul, We use 3 colors routinely for leukemia work with the CD45 v LSS gating technique. CD45 (PerCP or PE-CY5) is added to EVERY tube - leaving you the Fl1 and Fl2 detectors to add FITC and PE conjugated antibodies (so in effect we are doing 2 color typing and using the third as a gating antibody). You should read Michael Borowtiz's paper on CD45 v LSS for the background (Am J Clin Pathol 1993;100:534-540) and also get a hold of Greg Stelzer's paper in Annals of the New York Academy of Sciences (March 20 1993) - since this has the best "map" of where all the cell types fall (pp.268). Panels are always a contentious issue - I like the paper by the BCSH task force - J Clin Pathol 1994;47:777-781, but it's a little out of date now - remember to include the fashionable marker CD117 (see Bene etal Blood Vol 92 No 2 1998:596-599). However I don't like setting up a screening panel followed by a secondary panel since it involves double handling - and labour is the most expensive component of everything we do. So we have created "one shot" panels based on the published sources, with an emphasis on the patient population we see (mostly adult AML). Hope this helps. Peter Chapple Melbourne AUSTRALIA -----Original Message----- From: Paul Kuon [mailto:pmk3351@usl.edu] Sent: Thursday, September 09, 1999 9:17 PM To: cyto-inbox Subject: Acute leukemia How does one pursue an acute leukemia using three color analysis? How do you know where to look for the blast population, if the blasts are < 10% of the total? Do you use a CD45 Vs LSS histogram setup? We are new at this particular aspect and would appreciate any help available. Also any specific panels that are run for ALL ,AML,AMML etc. Thanks in advance. Reply to: pmk3351@usl.edu <mailto:pmk3351@usl.edu> or the group.
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