Dear Joy, We have found that a 1 (? Not sure off top of my head) mM level of EDTA helps, even with the Ca and Mg free medium. This was tried because we figured some of the clumping came via divalent ions released from dead and damaged cells arising during the dissection process. I would experiment with different EDTA (or citrate) concentrations in the divalent-ion-free medium, 0.1 through 2.0 mM (I think [Ca2+] can't get above 2mM, due to sulphate insolubility). [Mg2+] is much lower than [Ca2+] as well. Let me know, or get back to me for the exact details. Cheers, Bob. -----Original Message----- From: JOY MUNDY [SMTP:jmundy@immuno.imvs.sa.gov.au] Sent: Friday, July 11, 1997 2:50 AM To: Cytometry Mailing List Subject: Lymph Nodes that Clump Hi Can anybody help me with a cell suspension solution that will inhibit cell clumping. We disaggregate lymph node tissue for routine flow but have heaps of trouble with cell clumping. We have tried Mg and Ca free RPMI, also DPBS with DNAse to no avail. We keep blocking up the probe of the XL which is a bit of a bother. Willing to try anything!!!!!!! Joy Mundy Dept. Human Immunolgy Institute of Medical & Veterinary Science Adelaide, South Australia 5000 Ph 61-8-82287476; Fax 61-8-82324092 jmundy@immuno.imvs.sa.gov.au
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