Information from Jeff Harris related to this topic. Please contact him if you need further information. Marty >X-Webmail-User: jharris@itsa.ucsf.edu > >I don't know the references off the top of my head, but it is definitely >described that the sodium heparin green top tubes can activate >lymphocytes- that's why many labs use lavender/purple tops (using EDTA >as anticoagulant) or yellow top ACD (acid citrate/dextrose - citrate is >also an anticoagulant) tubes for culture/stimulation methods and even >phenotyping when you are worried about a labile marker - I must say >however, that for robust assays I don't think the heparin is a huge >effert, because we use them for the CFCs in the core lab and our >"non-stimulated" controls for cytokine secretion are quite negative, but >there is some small background activation by markers like CD69; similarly >we sort samples based on CD62L, which is also quite labile and sensitive >to stimulation/temperature/etc. but we try to use the blood freshly and >move quickly to separate the lymphocytes by Ficoll and keep them cold on >ice to avoid surface marker changes - whereas the core and clinical labs >use only EDTa-containing tubes for phenotyping for this reason. > > > >On Sat, 9 Mar 2002 19:55:35 -0800 Marty Bigos wrote: > >> Have you ever looked at this or anything similar? Marty >> >> >Date: Thu, 07 Mar 2002 17:03:50 -0400 (EDT) >> >From: MIGUEL-ANGEL PERALES <mperales@OPAL.TUFTS.EDU> >> >Subject: differences in blood collection >> >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >> >X-PMFLAGS: 34078848 0 1 18543.cnm >> > >> > >> >Not exactly a flow question, but somewhat relevant: We recently did a >> >study looking at T cell responses to a vaccine by Tetramer. Some >> >patients had green tops collected and others underwent leukapheresis. It >> >seemed that T cell responses were higher in green top samples. We didn't >> >compare the 2 methods in the same patient however. Has anyone had a >> >similar experience or tested this? Also, has anyone compared T cell >> >reponses in samples collected in green top vs, CPT tubes. >> > >> >Thanks, >> > >> >Miguel Perales >> >> > > > >Jeffrey M. Harris, M.D., Ph.D. >Asst. Adj. Professor, Pediatrics >University of California, San Francisco >Visiting Investigator >Gladstone Institute of Virology and Immunology -- Marty Bigos Director, Flow Core Gladstone Institute of Virology and Immunology Building 3 SFGH Rm 509 415-695-3832
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