[Histonet] iron hematoxylins[Scanned]

Wed Jun 15 08:52:49 CDT 2005

Heidenhain's uses sequence iron staining, as does Benda, Masson-Regaud
and Mallory. Heidenhain's differentiates in an iron alum solution,
Mallory in Ferric Chloride, Benda in acetic acid and Masson-Regaud in
alcoholic picric acid. This depends on mordanting with an aqueous ferric
salt solution, staining with haematoxylin until black, then
differentiating with an acid or a ferric salt until you visualise
cytologic detail.

Progressive or regressive iron haematoxylin staining (Janssens or
Weigerts) are similar to alum haematoxylin staining. Regressive staining
depends on an optimal amount of iron for dense staining (eg for the
demonstration of myelin), whilst progressive staining is made more
selective for nuclei by addition of acid or by an excess of ferric salt.

You see we are talking about different uses for iron haematoxylin, now I
wish I knew how ICC worked!!!  

-----Original Message-----
From: Bryan Llewellyn [mailto:llewllew <@t> shaw.ca] 
Sent: 15 June 2005 13:57
To: Histonet
Subject: Re: [Histonet] iron hematoxylins[Scanned]

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 
water.

Bryan Llewellyn

----- Original Message ----- 
From: "Rogerson Kemlo (ELHT) Pathology" <Kemlo.Rogerson <@t> elht.nhs.uk>
To: "Bryan Llewellyn" <llewllew <@t> shaw.ca>; "Histonet" 
<histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, June 14, 2005 11:40 PM
Subject: RE: [Histonet] iron hematoxylins[Scanned]

Confused????

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 [mailto:llewllew <@t> shaw.ca]
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
the
colour and I tried it to find out.  Blueing does deepen the colour,
although
not as significantly as with alum hematoxylin.  However, the acid
solutions
I used afterwards in Masson's trichrome undid it and the end result was
the
same.  It is useful if the iron hematoxylin is used alone without a
counterstain, perhaps on smears for amoebae, but otherwise not.  Other
iron
hematoxylin stains of the Heidenhain type do not need blueing.  The
mordant
does that.

Bryan Llewellyn

----- Original Message ----- 
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
To: "'Steven Coakley'" <sjchtascp <@t> yahoo.com>;
<Histonet <@t> lists.utsouthwestern.edu>
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
either
> black, or they aren't. If they are not, then the iron hematoxylin is
old
> (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: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Steven
> Coakley
> Sent: Tuesday, June 14, 2005 10:49 AM
> To: Histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] iron hematoxylins
>
> Does anyone know if using a bluing agent after weigerts iron
hematoxylin
> will achieve the same results a lengthy tap water rince?
>
> Steve
>
>
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