NIH-3T3 transfection studies

Dr. Wolfgang Schechinger Dr.Wolfgang.Schechinger at med.uni-tuebingen.de
Fri Sep 10 11:26:37 EST 1999


I made a good experience seeding 3T3 cells on coverslips placed in
polystyrene dishes without any further treatment. Either small ones
that one may remove for fixing, staining and examination or big ones
that are mounted directly in an observation chamber for living cells.
Sometimes it looked like the cells were "running" onto the glass
surface.

Coverslips were from GLW, Wuerzburg, 
big cover slips from Saur, Reutlingen (both: Germany)
PS cluster-dishes from TPP, Switzerland

...probably a bit far from the the UK and AU

If you need more details, please mail


Cheers and a sunny weekend!

Wolfgang









> From:          "A.F. Simpson" <AFS7 at le.ac.uk>
> Subject:       Re: NIH-3T3 transfection studies
> Date:          Fri, 10 Sep 1999 11:37:27 -0700
> Organization:  University of Leicester
> To:            methods at hgmp.mrc.ac.uk

> 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
> 
> love
> Anna
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touch! -----                              Dr. Wolfgang Schechinger  
Pathobiochemistry Dept.      University of Tuebingen, Germany email:
wgschech at med.uni-tuebingen.de * wwWait:
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usual disclaimers apply * This message is RNAse free - please don't touch!
-----                              
Dr. Wolfgang Schechinger   
Pathobiochemistry Dept.      
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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*unsolicited mail is *NOT* appreciated
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