CELL DEATH IN PLATES

Peter French p.french at ARNIE.CFI.UNSW.EDU.AU
Sun Nov 10 17:36:53 EST 1996


"angelpet" wrote:
>Rusnak is EXACTLY right.  It's an evaporation problem.  I use a lot of
>96-well plates and no longer use any of the outer wells . . . I fill them
>with sterile water.  So, I only get 60 wells, but I grow things long-term
>so . . . ..
>
>Don't be fooled by the name . . . I'm not a bimbo, but I play one on the
>net!
>
EXACTLY WRONG!
Sloppy thinking. Can we all get back to the original question (I reproduce
it here):
Thomas Kreuzer wrote:
> 
> Hello fellow netters,
> 
> I have been faced with a problem recently , for which I can find no solution
> altough it may be trivial.
> I seed HeLa - cells on standard 6-well cell culture plates (Costar) in 1 ml
> DMEM plus 5 % FCS , let them attach to the bottom overnight and then change
> the medium according to the tests I want to perform afterwards. I have noticed
> for some time that the cells in certain wells show a different behaviour than
> the others. For example: If I number the wells from left to right 1 to 3 in
> the upper row and 4 to 6 in the lower , the cells in number 4 are always more
> difficult to get off the bottom of the well with trypsin. First I thought it
> might be a handling problem during the tests , but last time I decided to let
> the cells grow for one additional day on the plate before the test , so I did
> not change the medium after the first night , but left it for further 24
>hours.
> When I looked through the microscope afterwards , I noticed that the cells in
> wells 1,2,3 and 5 had grown well , while in number 4 and 6 most of the cells
> were dead , altough an equal amount of cells had been attached to the bottom
> after the first night in all wells. The conditions were the same for all 6
> wells.
> So my question is: has anybody noticed a similar effect or has some
> suggestions why this might happen ?
> 
> Thanks in advance,
> Tom

A careful reading will show that it cannot be anything to do with
evaporation because:
1. It is a 6 WELL PLATE!!!! (not 96 wells as mistaken by the above writers)
2. This means that ALL WELLS ARE ON THE EDGE!!!


To me there are two possibilities. Either the batch of plates is faulty,
and therefore ask Costar for a different batch to try, or there is
something about the incubator that exists in that location - test this by
rotating the plate or moving it to a different spot in the incubator.

PLEASE, NO MORE EVAPORATION THEORIES

Peter French





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