From: Bruce Davis, MD - Clinical Pathology (bdavis@beaumont.edu)
Date: Fri Jun 09 1995 - 11:15:43 EST
My experience with neutrohil activation markers, which generally involve an entire population change, rather a subset or % postive phenomenon, has been recently published in Laboratory Hematology 1:3-12, 1995 (a new journal published by the International Society for Laboratory Hematology and edited by Ken Ault, M.D.). We have been quite satified with using Quantum FITC or PE beads or simply cellular beads from Flow Cytometry Standards, Corp. along with Quickcal for Winlist (Verity Software House). The analysis is relatively simple and can be expressed in meaningful units (eg. FITC equivalents or antibody binding sites). Bruce H. Davis, M.D. Dept. of Clinical Pathology William Beaumont Hosp. Royal Oak, MI 48073 810-551-5137 FAX 810-551-8057 On Thu, 8 Jun 1995, James Weaver 301-594-5879 FAX 301-594-3037 wrote: > Uno Johansson posted: > > > Hi everybody! > > Does anyone have experiences or references concerning mathematical handling > > of data from surface activations markers on lymphocytes using FACS? Any > > suggestions would be greatly appreciated. Thank you very much. > > An excellent place to start is: > > Flow cytometric titration of retroviral expression vectors: Comparison of > methods for analysis of immunofluorescence histograms derived from cells > expressing low antigen levels. Sladek, T.L. and Jacobberger, J.W. Cytometry 14 > 23-31 (1993). > > >From my limited evalution of CD69 as a proliferation marker (not published) I > found the issue complex. From discussions I have had with others I am not alone > in this response. In brief, If markers go from absent to bright such as CD25, > then the evaluation can be done on % in the 'bright' area of the histogram. On > the other hand, if the marker goes from dim to bright in a continuously changing > > fashion such as CD69 then the evaluation becomes difficult. This requires curve > fitting that can handle asymmetric peaks such as a Weibull function. ModFit from > > verity has such a function but it does not recognise Log scale data. Since it > was intended for DNA analysis, it assumes that all data is linear. If there are > other analysis packages that will do Weibull fitting on Log scale data, I am not > > aware of them. If there has been further progress that I am not aware of I would > > be interested in hearing about it myself.This is a complex area and I wish you > the best of luck. > > > > +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ > James L. Weaver Email: WEAVER@FDACD.CDER.FDA.GOV > Molecular Pharmacology Phone: 301-594-5879 > Division of Research & Testing FAX: 301-594-3037 > ORR, CDER, Food & Drug Admin. Voice: 'Hey you' > Laurel, MD > +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ > > >
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