RE: [Histonet] tissue coming off slides

From:"Mark Elliott"

I coat slides with celloidin for immunos here with little problem.  I
find some of the higher pH retrieval methods cause lose of tissue so
need the coating.  I just extend the time in each of the reagents by
half and get nice staining.
Mark Elliott
Vancouver, BC

>>> "Monfils, Paul"  10/05/06 11:16 AM >>>
There is the venerable old technique, seldom used today, of coating
slide with celloidin, also known as parlodion. The technique isn't used
today, partially because electrostatically charged slides have solved
of the adherence problems, and partially because the technique
ethyl ether, which is forbidden by many institutions today because of
extreme flammability.  But what we used to do is make up a solution of
celloidin in 1:1 ethanol/ethyl ether, deparaffinize the slides through
absolute alcohol, immerse them in the celloidin solution for about a
then transfer them to 80% ethanol for a few minutes to harden the
film. After that they can be rehydrated and stained as usual. The
forms a continuous semipermeable membrane over the entire slide and
which physically holds the section in place during staining. Most dye
molecules can pass through the celloidin membrane and stain the
Antibody and enzyme molecules are too large to pass through the
membrane, so
techniques involving them cannot be done.  Some standard histochemical
procedures will stain the celloidin as well as the tissue, but this is
usually a problem because during the final dehydration prior to
coverslipping, the xylene will dissolve away the celloidin.  Many years
we used this technique particularly for silver stains involving
since the ammonia would attack the gelatin we used as a section
and cause the sections to fall off the slides.

> ----------
> From: on behalf of
> Baker
> Sent: 	Thursday, October 5, 2006 9:00 AM
> To: 	Histonet
> Subject: 	[Histonet] tissue coming off slides
> Hi All,  A group I work with just came to ask if there was a way of 
> keeping frozen tissue sections from coming off a slide?  It seems
> technician used uncoated/regular slides to mount 100's of sections. 

> The tissues were embedded in OCT and cut at 5 microns but of course
> sliding off during any staining procedure.  Is there a protocol of
> sort to coat the slides after the fact to retain the sections through

> staining?  I at least suggested they stain flat.  Any suggestions are

> appreciated.  John
> John A. Baker
> The University of Michigan
> Orthopaedic Research Laboratories
> Histology Unit
> 109 Zina Pitcher Place, 2218 BSRB
> Ann Arbor, MI 48109-2200
> 734-936-1635
> _______________________________________________
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