Re: Tissue sections on slides

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From:"Richard G. Lea" <rgl@rri.sari.ac.uk>
To:"John C. Dennis" <dennijc@vetmed.auburn.edu>
Reply-To:
Date:Wed, 11 Aug 1999 17:44:35 +0100
Content-Type:text/plain; charset=us-ascii

John C. Dennis wrote:
> 
> Richard
> 
> Are you using any EtOH in treating your tissue before you mount the
> sections on slides?
> We were having a similar problem just the other week.  Our frozen
> sections were coming off the
> slides poly L lysine or no.  The tissue had been fixed but held in 70%
> EtOH until sectioning.
> Once I stopped exposing the tissues to EtOH the sections stopped coming
> off the slides.
> 
> John Carroll Dennis
> Anatomy, Physiology, and Pharmacology
> 109 Greene Hall
> Auburn University, AL  36849
> 
> On Wed, 11 Aug 1999, Richard G. Lea wrote:
> 
> > To whoever can help,
> >
> > I am attempting to carry out in-situ hybridisation on sections of tissue
> > which have been cultured for 24 hours.  The control non-cultured tissues
> > are fine but after culture the tissue sections fall off the slides
> > during the in-situ protocol.  We have tried TESPA and double coated
> > gelatin, PLL slides however the problem remains.  How can I stop these
> > sections coming off the slides - any suggestions gratefully received.
> >
> > Richard Lea
> > --
> > Richard G. Lea PhD
> > Senior Research Scientist
> > Division of Nutrition, Pregnancy and Development
> > Rowett Research Institiute
> > Greenburn Road
> > Bucksburn
> > Aberdeen AB21 9SB
> >
> > Tel:  44(0) 1224 712751 x 2319
> > Fax:  44(0) 1224 716622
> >

John,

Many thanks for your reply.  We are using paraffin sections cut from  
Bouins fixed tissues which are stored in 70% Et-OH until standard
processing.  We have to dehydrate through an ethanol series for this and
we have optimised the fixation for our in-situ hybridisation protocol.  

I think we have to look at alternative slide coatings or use
electrostaticaly charged slides to stop our cultured tissue sections
coming off the slides - do you have any suggestions as to which may be
the most effective ?

-- 
Richard G. Lea PhD
Senior Research Scientist
Division of Nutrition, Pregnancy and Development
Rowett Research Institiute
Greenburn Road
Bucksburn
Aberdeen AB21 9SB

Tel:  44(0) 1224 712751 x 2319
Fax:  44(0) 1224 716622



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