AW: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue

[Krista Sider]

Hi Gudrun

How deep would you consider the oxidation to go? I have tried samples from
around 200um into the block. The Alkaline Alcohol converts the alcian blue
to monastral fast blue, which is insoluble - this is what i have been told.
So locks it in I guess. Doesn't really matter as I don't get alcian blue in
the sample to lock in.

Thanks
Krista
On May 28, 2013 10:09 AM, "Gudrun Lang" <gu.lang <@t> gmx.at> wrote:

> Hi Krista,
> Maybe the bad stainability is more due to oxidation by air of the old
> block-surface than due to fixation. A deeper slide for comparison would
> show.
> I don't know what the alkaline alcohol is good for ?
>
> Gudrun
>
> -----Ursprüngliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Krista
> Sider
> Gesendet: Montag, 27. Mai 2013 23:28
> An: histonet <@t> lists.utsouthwestern.edu
> Betreff: [Histonet] Movat Pentachrome - Need to enhance Alcian Blue
>
> 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
> Krista
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