NIH-3T3 transfection studies

A.F. Simpson AFS7 at
Fri Sep 10 13:37:27 EST 1999

alex dobrovic wrote:
> Hi
> We are doing NIH-3T3 transfection studies with a GFP fusion protein that we
> want to visualise under confocal microscopy. We have had a lot of trouble
> getting the cells to stick down on glass slides or coverslips. We would be
> grateful for any suggestions.

Have you tried coating the glass with something to make it more
cell-friendly?  Poly-l-lysine, gelatine or diluted Matrigel might be
things to try.  Sometimes even just incubating the slides/coverslips
with FCS for thrity minutes or so then giving a quick rinse in PBS
before plating the cells will help.  I have no idea if any of these
methods will interfer with your GFP visualisation, though - perhaps
someone else does?

You could also try changing the brand of slide you use - some cells just
don't like a particular brand.  We use Lab-tek II chamber slides from
Nalgene which are quite satisfactory.

> Alex


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