Charles, I have begun doing cytoplasmic staining on CLL patients that do not exhibit surface light chains. I use the stained and fixed tube, assuming there is enough left after acquisition. I add 100 ul reagent B from Caltag's fix and perm and more light chain antibody and incubate for 15 minutes. Then I wash in PBS, resuspend, and acquire again. Quite often the light chains appear! Thanks for the CD 38 reference. I'll definitely read that. Melanie O'Donahue Mayo Clinic Scottsdale -----Original Message----- From: Repetti, Charles [mailto:crepetti@aml.com] Sent: Monday, December 11, 2000 12:59 PM To: cyto-inbox Subject: RE: Several Questions Melanie; The CD38 references are in Blood, Sept 15, 1999Vol 94,pp1837-1839; pp1840-1847 (the flow CD38); pp 1848-1854. CD5-FITC, CD38-PE, CD19-PE:CY5. The CD38 is usually discreet, almost all negative or mostly positive in the 2nd or 3rd decade. CLLs without K,L: have very little experience with intracellular, would recommend intsead having alternative k,l clones for surface staining. Would be interested in any info on cytoplasmic stains. good luck Charles -----Original Message----- From: O'Donahue, Melanie . [mailto:ODonahue.Melanie@MAYO.EDU] Sent: Friday, December 08, 2000 4:36 PM To: cyto-inbox Subject: Several Questions Hello all, There are several things I'm trying to troubleshoot and/or improve and I would appreciate any input from my fellow flowers. Are any of you doing platelet hyper aggregation studies with flow? Also, have you had any experience running CD 38 on CLL patients? It's supposed to be a prognostic indicator. One of our heme onc docs is interested in it. On CLL patients that don't express light chains, do you routinely run them intracellularly? What rate of success do you have with that? Final question. Do you run your light chains in two separate tubes? Is their a reason for that? We are having problems with our three color (kappa-lambda-CD 19) tube and I'm wondering if I should be running two color instead. Thanks for any input. Have a great weekend! Melanie O'Donahue, MT, ASCP Mayo Clinic Scottsdale
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