brett at BORCIM.WUSTL.EDU
Wed Dec 2 12:39:47 EST 1998
>In article <366400DA.759D048 at sghms.ac.uk>, Claire Fenech
><c.fenech at sghms.ac.uk> wrote:
>>> Hi all,
>>> i was wondering if anyone can get rid of cell clumping together after
>>> trypsining them of the tissue culture dish?
>>> I normally can get them of the tissue culture dish no problems but the
>>> cells still sticks to gether making them hard to count. is there a good
>>> way to get over this problem.
>>If your cells are not too fragile you can resuspend them in fresh medium
>>(small amount 1,2 ml) and "triturate" them gently: sucking them up and down
>>a sterile, fire polished glass pasteur pipette. Keep the tip of the pipette
>>close to the bottom while you draw or expel the cells. This dissociate the
>And for even more stringent trituration which virtually guarantees that over
>90% cells are single, you can jam sterile yellow tip on the 5-10 ml pipet
>and do ~ 10 passes. Works great (kills some % of cells though).
>Other suggestions also apply - split before confluent and trypsinize
> - Dima
Actually, the best way I have found is to buy some of those sterile disposable
plastic sieves that fit perfectly into the top of a 50 ml Falcon conical. They
will retain large clumps and be broken up by showering the residue with media
to release additional single cells. Dima's plan sounds too barabaric to me.
brett at borcim.wustl.edu
Dept of Molecular Microbiology
Washington University School of Medicine
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