Re: [Histonet] iron hematoxylins[Scanned]

From:Bryan Llewellyn

Iron hematoxylins of the Heidenhain type do not use acids and usually no 
counterstain is applied.  That type uses three bath staining.  The first is 
mordanting in a ferric compound, usually ferric ammonium sulphate, the 
second is in the hematoxylin, and the third usually returns to the mordant 
to differentiate.  Acids are not usually involved, and the third step leaves 
the tissue black stained.  An exception is some modifications which replace 
the mordant with an acid, but these are not commonly used.  Extensive 
washing is recommended with them, but to remove traces of mordant rather 
than to alkalise, although that may happen depending on the pH of the tap 

Bryan Llewellyn

----- Original Message ----- 
From: "Rogerson Kemlo (ELHT) Pathology" 
To: "Bryan Llewellyn" ; "Histonet" 

Sent: Tuesday, June 14, 2005 11:40 PM
Subject: RE: [Histonet] iron hematoxylins[Scanned]


Isn't haematin like litmus and changes colour in acid and alkali
solutions? Doesn't the mordant act as a 'key' to the proteins and
haematin causing the dye to 'bind'? So why, if the iron haematoxylin is
in an acid 'environment' does it not require 'blueing'?

-----Original Message-----
From: Bryan Llewellyn []
Sent: 15 June 2005 02:29
To: Histonet
Subject: Re: [Histonet] iron hematoxylins[Scanned]

I have blued Weigert's iron hematoxylin in the past, although I didn't
do it
as a routine.  I wondered if the acid in the Weigert's solution affected
colour and I tried it to find out.  Blueing does deepen the colour,
not as significantly as with alum hematoxylin.  However, the acid
I used afterwards in Masson's trichrome undid it and the end result was
same.  It is useful if the iron hematoxylin is used alone without a
counterstain, perhaps on smears for amoebae, but otherwise not.  Other
hematoxylin stains of the Heidenhain type do not need blueing.  The
does that.

Bryan Llewellyn

----- Original Message ----- 
From: "Lee & Peggy Wenk" 
To: "'Steven Coakley'" ;

Sent: Tuesday, June 14, 2005 5:07 PM
Subject: RE: [Histonet] iron hematoxylins

> I've never blued after a Weigert iron hematoxylin stain. Nuclei are
> black, or they aren't. If they are not, then the iron hematoxylin is
> (more than a couple days old), and putting the tissue into an alkali
> solution won't bring the color back.
> I'm curious, why are you bluing? What is the purpose? Maybe then I can

> help.
> Peggy Wenk, HTL(ASCP)SLS
> William Beaumont Hospital
> Royal Oak, MI 48073
> -----Original Message-----
> From:
> [] On Behalf Of Steven
> Coakley
> Sent: Tuesday, June 14, 2005 10:49 AM
> To:
> Subject: [Histonet] iron hematoxylins
> Does anyone know if using a bluing agent after weigerts iron
> will achieve the same results a lengthy tap water rince?
> Steve
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