[Histonet] Movat Pentachrome - Need to enhance Alcian Blue

Krista Sider krista.sider <@t> utoronto.ca
Mon May 27 16:28:06 CDT 2013

Hi All,

I had wonderful help from this site for my Woodstain Scarlet-Acid Fuchsin
binding collagen problem. Now that this is fixed I have realized that the
Alcian Blue is not staining my excessively fixed samples (Human Aorta &
Aortic Valve, formalin fixed 2-4 years, embedded in paraffin and stored at
RT for ~6 years). I have some samples fixed for only 2-6 months and these
are fine but the samples fixed for 2-4 years do not uptake the Alcian Blue.
I have taken it up to 1 hr in the 1% Alcian Blue solution followed by 10
min in 56*C Alkaline Alcohol solution. I do a 1hr Bouin's 50*C to start (I
have also tried Bouin's at 60*C).

Any ideas on how I can get the Alcian Blue to stain these excessively fixed
samples or possibly a substitute? I need to see the proteoglycans/GAGs.

Thank you

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