[Histonet] need help for epon

[J. A. Kiernan]

You would not expect a trichrome method for
paraffin sections to work properly after
glutaraldehyde, osmium tetroxide and epoxy
resin embedding. Both the fixatives
interfere with amino groups (to which the
dyes in a trichrome are attracted, and
both glutaraldehyde fixation and the
cross-linked resin interfere with 
penetration of the larger dye molecules
in the mixture.

5um is thicker than usual for such sections. 
If you cut at 0.5 or 1 um you will see all 
the structural details  with a one-colour 
stain such as alkaline toluidine blue. The 
staining intensity can be increased by 
first putting a drop of concentrated sulphuric 
acid on top of the section for about 20 
seconds then rinsing  off with tap water 
before staining.  
-- 
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan <@t> uwo.ca
   http://publish.uwo.ca/~jkiernan/
______________________________________
Myri37 <@t> aol.com wrote:
> 
> hello
> i have embedded thin tissue in epon, after postfixation with
> osmium
> i stuck sections of 5 um on slides with haupt's solution,
> deplastized in ethoxide of sodium in heat for 30 minutes
> and after that stained Goldner's trichrome
> tissue was  not coloured, and i dont understand why, is it
> because of osmium ?
> pleaze do you have a protocol of Goldner's trichrome especially
> for epon not for parrafin sections, or another stain to show
> collagen and mineralized tissu ?
> Myriam
> Natural implant
> France

-- 
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan <@t> uwo.ca
   http://publish.uwo.ca/~jkiernan/