[Histonet] H&E+ Alcian Blue

[Deborah Faichney]

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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