From: Bakul Dalal [Dr.] [VH] (bdalal@vanhosp.bc.ca)
Date: Thu Nov 20 2003 - 17:15:14 EST
We use whole blood, within 12 hours of collection, stain for 55, 59 and FLAER, run a healthy control, gate by SS / FS, look at RBC, PMN and monos. We call it positive if two markers are absent in two cell lines. We also report whether the abnormality is type II or type III. We also quantitate the clone using PMNs and RBCs using CD59, and report it as % Hope this helps. Bakul I. Dalal MD FRCPC FCAP FASCP Director, Flow Cytometry Laboratory Assoc Director, Hematology Laboratory Vancouver General Hospital Clinical Professor, Faculty of Medicine University of British Columbia bdalal@vanhosp.bc.ca 604 875 4496 / 4633(f) This is the best day the world has ever seen. Tomorrow will be better. R. A. Campbell >>> <MailingList/general/Biocytex@biocytex.fr> 2003-November-19 2:26:53 AM >>> Dear Sara, May I suggest you to have a look to the following papers : 1/ OELSCHLAEGEL U., et al. (2001) A standardized flow cytometric method for screening paroxysmal nocturnal haemoglobinuria (PNH) measuring CD55 and CD59 expression on erythrocytes and granulocytes. Clin.Lab.Haem. 23, 81-90. 2/ HSI E.D., (2000) Paroxysmal nocturnal hemoglobinuria Testing by flow cytometry. Am J Clin Pathol, 114, 798-806. 3/ PONCELET P. et al. (2000) Clinical applications of flow cytometric immunophenotyping in acute lymphoblastic leukemia, in : C.C. Stewart & J.K.A. Nicholson (eds) Immunophenotyping, Wiley-Liss, pp161-180. I know other approaches than using antibodies have also been proposed, see for example : BRODSKY, R.A. et al. (2000) Improved detection and characterization of paroxysmal nocturnal hemoglobinuria using fluorescent aerolysin. American Journal of Pathology, 114, 459-466. Hope that helps Philippe Poncelet, PhD Director, Research and Technology BioCytex 140, Chemin de l'Armée d'Afrique 13010 Marseille France Tel: +33 (0) 4 96 12 20 40 Fax: +33 (0) 4 91 47 24 71 www.biocytex.fr
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