[Histonet] Masson's Trichrome, Weigert Hematoxylin Equivalent

John Kiernan jkiernan <@t> uwo.ca
Wed Mar 10 21:04:41 CST 2010

No. The PTA or PMA used in Masson's method is too acidic and would remove Harris's or any other haemalum. An iron-haematoxylin such as Weigert's or Lillie's gives an acid-resistant nuclear stain that is progressive and almost black. 
The stock solutions for Weigert's are good for many years. The mixture keeps for 3 or 4 days at room temp or 10-14 days in a fridge. Lillie's modification is more stable - OK for 2-3 weeks at room temperature. These iron-haematoxylins are ready to use as soon as they are mixed. No "ripening" is needed because the ferric chloride serves as both the oxidizing agent and the metal salt that forms a black complex with haematein. Deterioration with time is due to over-oxidation, converting haematein to other compounds. If the nuclei have a brownish shade instead of clean blue-black it's time to make a new working solution. 
John Kiernan
Anatomy, UWO
London, Canada
= = =
> ----- Original Message -----
> From: cscampbe <@t> uci.edu
> Date: Wednesday, March 10, 2010 19:21
> Subject: [Histonet] Masson's Trichrome, Weigert Hematoxylin Equivalent
> To: HistoNet <histonet <@t> lists.utsouthwestern.edu>
> > Hi Histonet,
> > 
> > I am performing the trichrome stain on cardiac tissue. The 
> > procedure calls
> > for Weigert's iron hematoxylin stain or equivalent. Would Harris
> > hematoxylin work as well. It's what I have readily available at 
> > the lab.
> > 
> > The instructions for preparing the Weigert stain involves mixing fresh
> > solutions from two stock solutions - but the staining process of 
> > all the
> > slides will take weeks. How long does the solution stay "fresh?"
> > 
> > Thanks!
> > -Colin
> > 
> > 
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