We had a client that used to run lymph tissue and whole tumors that he had deparafinized. Talk about messy samples! The thing that worked for him was to first filter through 50 micron mesh to get rid of really big clumps and then, right before the sample went on the cytometer, to run the sample up and down through a small gauge needle with a turberculin syringe. This must be done slowly and gently as to not shear all the cells. We have also had succes with this method on other whole tissues such as brain, liver, and a number of plant tissues. Happy flowing. David McFarland University of Illinois Biotechnology Center Flow Cytometry Facility ---------- Forwarded message ---------- Date: Thu, 10 Jul 1997 16:49:52 GMT+1030 From: JOY MUNDY <jmundy@immuno.imvs.sa.gov.au> To: cyto-inbox 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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