96 well - glass bottom dishes - glass unadheres

[Dan Tom]

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Hi List,

A user from my facility is having a difficulties staining cells on 96 well
glass bottom plates.  The glue that adheres the glass to the dish dissolves
during the staining process causing the media to leak out.

The user has fixed the cells with and without methanol, which the
manufacturer indicates as having good compatibility, and they also tried
paraformaldehyde.  Each time, the glue dissolves.  The current working
hypothesis is that the refrigeration process might affected the adhesion.

Could someone with a similar experience suggest what we can be doing wrong?
 I'm sorry I don't have any more details.  Any suggestions would be appreciated.

Thanks,
Dan

=========================

Daniel Tom

Harvard NeuroDiscovery Center

Optical Imaging Facility

220 Longwood Ave.

Goldenson Bldg., rm. 501

t: 617-432-1442

[Csúcs Gábor]

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Dear Tom,

Of course the effect can be brand dependent. Nevertheless, we had so far
no problem with our glass bottom plates, when we fixed the cells with
paraformaldehyde. Keeping the fixed and stained plates at 4 degrees had
also no effect. I can imagine that if you put them into -20 or - 80
(although I don't know why someone would do this...) then  the differen
thermal expansion of the glass and plastic may cause problems.


Cheers   Gabor

[Sylvie Le Guyader-2]

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Hi Dan

You could try to fix your cells, remove the wells and the leftover glue then
circle your cultures with a PapPen (Hydrophobic) then do your staining. It
works very well and you would need far less reagents than with the wells.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
[phone]

> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Dan Tom
> Sent: 12 August 2008 22:31
> To: [hidden email]
> Subject: 96 well - glass bottom dishes - glass unadheres
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi List,
>
> A user from my facility is having a difficulties staining cells on 96 well
> glass bottom plates.  The glue that adheres the glass to the dish

dissolves
> during the staining process causing the media to leak out.
>
> The user has fixed the cells with and without methanol, which the
> manufacturer indicates as having good compatibility, and they also tried
> paraformaldehyde.  Each time, the glue dissolves.  The current working
> hypothesis is that the refrigeration process might affected the adhesion.
>
> Could someone with a similar experience suggest what we can be doing
wrong?
>  I'm sorry I don't have any more details.  Any suggestions would be
appreciated.

>
> Thanks,
> Dan
>
> =========================
>
> Daniel Tom
>
> Harvard NeuroDiscovery Center
>
> Optical Imaging Facility
>
> 220 Longwood Ave.
>
> Goldenson Bldg., rm. 501
>
> t: 617-432-1442