clumping and sieving

Carolyn Jefferiss prscmj at bath.ac.uk
Tue Sep 19 07:23:39 EST 2000


Dear All,

Has anyone tried using "Accumax" from TCS Cellworks at all in flow
cytometry? It is supposed to be an anticlumping reagent, recommended
for using for cell counting, which requires little or no Ca++ or
Mg++. The cells are not suitable for culturing after disaggregation
in this reagent. Anyone?

My little bit on clumping on the sorting front;
We sort cultured marrow cells which can be horribly clumpy and sticky
(hence my wondering about Accumax) and we sort with a suspension at
10x10e6cells/ml having filtered the cells immediately before putting
on the machine and the proportion of dead cells definitely affects
the amount of clumpiness which develops. We don't put on more than
2ml at a time, and during a run take off the tube and give it a very
gentle mix if it is really settling out, rather than using the buzzer
on the machine. They can always get a second pass through a filter.
The on-line thing sounds like a big faff if you get it clogged up so
we've never done it. Since persuading everyone of the need to filter,
before running each and every sample, we haven't had ANY blockages.
(Famous last words).

Carolyn Jefferiss

Carolyn Jefferiss Ph. D.
Pharmacy and Pharmacology
University of Bath
Claverton Down
Bath BA2 7HY



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