RE: Lymph Nodes that Clump

Bob Ashcroft (
Fri, 11 Jul 1997 09:55:47 +1000

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 []
Sent: Friday, July 11, 1997 2:50 AM
To: Cytometry Mailing List
Subject: Lymph Nodes that Clump


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

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