[Histonet] H&E+ Alcian Blue

[Bryan Llewellyn]

You are probably using a regressive hematoxylin.  Most of those stain acid 
mucins purple-blue.  To overcome it use a strictly progressive hematoxylin 
such as Mayer (hx 1 g., alum 50 g.) for 5 minutes.  The order should be 
alcian blue, wash, H&E.

Bryan Llewellyn


----- Original Message ----- 
From: "Deborah Faichney" <d.a.faichney <@t> stir.ac.uk>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, September 09, 2010 1:44 AM
Subject: [Histonet] H&E+ Alcian Blue


Hello all,

I have a request to carry out a combined staining with H&E + Alcian Blue 
pH2.5.    I have tried in vain to get this to work but regardless of the 
order of staining the end result is dark blue/purple mucin.  I have carried 
out a parallel experiment whereby the staining has been checked 
microscopically then stopped after each of the dyes.   (Thus giving 3 slides 
stained with:  AB, AB+H and AB+H+E)  The AB and AB+H are really nicely 
stained but as soon as the eosin is added (using 2 different stains and a 
variety of times) the mucin staining looks similar to the nuclear stain.  I 
am expecting the alcian blue to remain turquoise: should it?

For information, I am trying to stain salmon intestine at 5 microns for 
using the following:

Alcian Blue 8GX (certified), pH has been checked
Haematoxylin Z (Cellpath Uk)
1% aq Eosin (Cellpath uk) and lab prepared solution from dye.

Thanks from a frustrated technician!

Debbie Faichney
Histopathology
Institute of Aquaculture
University of Stirling
Scotland
UK








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