Alcian Blue/Alcian Yellow stain

From:Michelle McDonald

Alcian Blue/Alcian Yellow stain

Hi all,

I am having trouble with a combined Alcian blue Alcian Yellow stain with Saffranin O counterstain protocol. The Alcian Blue is at pH 1.0 and the Alcian Yellow at pH 2.5. The Saffranin O is a 0.5 % aqueous solution. I am using the stain for cartilage analysis.

After the stain is complete I get very nice contrast between the yellow and blue  with a pale red background..... however when I dehydrate the sections through alcohols for mounting I lose all yellow staining, the saffranin  O  red replacing it. When I try mounting with an aqueous mountant the Saff O and Alcian yellow bleed out of the sections completely. I have tried air drying them and then mounting in Eukitt and running through xylenes only before eukitt... all to no avail.

If anyone has any suggestions I would really appreciate them as this stain is showing great analytical potential for my work.

Thanks in advance


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