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Dispersing big cell clump growing in flask - Big cell clump in flask (Apr/02/2007 )


How do get rid of a large cell clump with central necrosis growing in the flask that was probably the result of inadequate dispersion in the previous passage?

My cell dispersion during passage is also not good. This has also affected my seeding in the 6 well plate
I've tried pipetting the cell solution many times, trying different small sized pipettes, placing the pipette tip against the wall of the flask during dispersion ( prob injured cells in the process too).
Am working with Hep 3B cells which are large and so even though the field looks confluent, there aren't many cells. When I seed into a 6 well plate, it often happens that there will be a few large nests clustered together in a few areas whereas the other areas are devoid of cells.

Any suggestions? Would be very grateful since it's messing up my transfection too. ph34r.gif


-A cell-

you may wish to check this post.
for some useful information.

-Minnie Mouse-