clumping and sieving
C. Kevin Becker
ckb at phnxflow.com
Tue Sep 19 19:04:52 EST 2000
While TCS CellWorks is our very capable distributor in the UK, Accumax (and
its sister product Accutase) are manufactured by Innovative Cell
Technologies, Inc. (a Phoenix Flow Systems company based in San Diego). Our
protocol for using Accumax for tissue dissociation for primary cell culture
calls for the tissue to be placed in Accumax for up to 25 minutes. The
viability following this treatment is typically very good. We have several
customers who use Accumax to disaggregate "clumpy" cell lines which will
then be run on a cell sorter. Again, viability is typically very good. We
are confident that the use of this product will very likely reduce the
problems Dr. Jefferiss is having with the cultured bone marrow cells,
without having a serious adverse effect on viability. We are happy to work
with any of our customers on optimizing protocols for these types of
applications. Please visit our website (www.innovativecelltech.com) for
additional information on these products.
Phoenix Flow Systems, Inc./Innovative Cell Technologies, Inc.
----- Original Message -----
From: Carolyn Jefferiss <prscmj at bath.ac.uk>
Sent: Tuesday, September 19, 2000 5:23 AM
Subject: clumping and sieving
> 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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