[Histonet] Mucin containing cells too blue

[Bryan Llewellyn]

If you go to their web site and look at the MSDS for the hemalum it appears 
to contain about 6 grams hematoxylin per litre, with both alum and aluminum 
sulphate for mordant, and hydrochloric acid (about 2 mL per litre).  It 
comes 80% oxidised when fresh, if the amount of sodium iodate is anything to 
go by.  In other words, a very strong hemalum with full oxidation and 
hydrochloric acid as pH control.  I'm more surprised that it doesn't always 
stain mucin, frankly.

Try adding more hydrochloric acid.  It is probably the pH is too high. 
Mucins stain around pH3-4.  Add hydrochloric acid to get it below that. 
Alternatively, and a MUCH better idea, use a hemalum with about 3 grams 
hematoxylin per litre.  It is much less likely to stain mucin.  To do that, 
make up a solvent containing everything in the hemalum except the 
hematoxylin and oxidant, then dilute equal parts.  Or, try Coles with acid.

Bryan Llewellyn


----- Original Message ----- 
From: "Greg Dobbin" <gvdobbin <@t> ihis.org>
To: <Histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, January 22, 2008 12:24 PM
Subject: [Histonet] Mucin containing cells too blue


> Hi Folks,
> I have been using the Surgipath Selectech reagents (H&E) for almost 3
> months now and I have not had a single complaint about staining...until
> now.  Suddenly the mucin containing cells are too blue.  My first
> thought was to make fresh Define (the Surgipath differentiator).  This
> did not fix the problem.  Next I replaced the hematoxylin with fresh
> (not supposed to have to change before 2500 slides, but changed at 1500
> slides to see if that would correct the problem). It has not.
>
> I was sure that fresh hematoxylin (and thererfore correct pH) would
> rectify the problem.  What are the other usual suspects here?  I should
> also mention our H&E's are done on the Leica Autostainer.
> I look forward to receiving your collective insight!
> Greg
>
>
>
> Greg Dobbin, R.T.
> Chief Technologist, Histology Lab
> Dept. of Laboratory Medicine,
> Queen Elizabeth Hospital,
> P.O. Box 6600
> Charlottetown, PE    C1A 8T5
> Phone: (902) 894-2337
> Fax: (902) 894-2385
>
> There is some merit in doing the right thing rather badly,
> but absolutely none in doing the wrong thing excellently!
>
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