a problem in making the stable cell line of a suspension cell

Emir Khatipov khatipovNO at NOuchicago.edu
Wed Oct 2 19:25:58 EST 2002


It is also possible to use some dyes that bind to dead cells to sort those
from live ones using flow cytometry (FACS). I cannot give you any specifics
right now, but from what I recall, there are DNA-binding dyes that would not
penetrate live cells. There is also Trypan blue dye that could be used for
the same purpose, except that it is not a DNA binding dye, but just binds to
cytoplasmic proteins (=dead cells).
Emir




"Nick Theodorakis" <nicholas_theodorakis at urmc.rochester.edu> wrote in
message news:3d9b89a7.5922382 at netnews.worldnet.att.net...
> On 2 Oct 2002 15:20:45 -0000, linglingyang at yahoo.com wrote:
>
> >Hi, guys. I have a problem in making the stable cell line of a suspension
cell (U266). Despite of optimization by electroporation and other methods,
its transfection efficiency is low. So how can I separate the transfected
live cells from hundreds of dead cells around them? Only wait for the live
cells to grow and out number the dead cells? The U266 grows pretty slow,
though. What a headache!
> >
>
> I suppose you could cotransfect with a GFP plasmid and FACS sort them.
>
> Nick
>
> --
> Nick Theodorakis
> nicholas_theodorakis at urmc.rochester.edu





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