CELL DEATH IN PLATES
herro001 at maroon.tc.umn.edu
Thu Nov 14 07:51:43 EST 1996
It really does sound like a bad batch of plates. Perhaps you could post
the lot number and people with the same lot could compare notes?
Peter French wrote:
> "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
> 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
> > 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
/ Michael J. Herron, U of MN, Dept. of Dermatology /
/ herro001 at maroon.tc.umn.edu /
/ 612-625-8935 Box 98 UMHC, Mpls MN 55455 /
/ http://RASH.med.umn.edu/ /
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