[Histonet] Trichrome staining and fibrin

Bryan Hewlett bhewlett <@t> cogeco.ca
Tue Jun 13 11:13:57 CDT 2006


Actually NOT!
True, Lendrum's original papers(1962) recommended fixation in picro-mercuric 
alcohol for 3 weeks (a tad unnecessary that!).
However, both Prof. Lendrum and Bill Slidders also successfully used 
formaldehyde fixed material that was treated in Bouin immediately prior to 
The key to all of the Masson Type trichromes on formaldehyde-fixed tissue, 
is to use a pretreatment in picric acid.
This re-aligns the reactive side chains on the proteins, so that there is a 
predominance of basic amino groups and hence maximal binding of anionic 
Mercury fixation is simply not necessary! Nor is Zinc substitution. (see 
Troubleshooting Histology stains: Horobin and Bancroft 1998, page 129)
The lack of consistency, in obtaining good red colour, is due variability in 
the staining procedure (the post dye water rinses), NOT the fixation.
Once this is addressed, the inconsistency goes away.
That is true for the original Masson, Picro-Mallory (all variants), MSB and 
Masson 44/41.
Achieving consistent good red colour with formaldehyde fixed material is 
easy, if the staining mechanisms are understood and those unnecessary water 
rinses removed.
Been doing it for over 40 years!

(Engineers can prove that the bumble bee simply cannot fly. However, the 
bumble bee doesn't know that and flies anyway)

----- Original Message ----- 
From: "Marshall Terry Dr, Consultant Histopathologist" 
<Terry.Marshall <@t> rothgen.nhs.uk>
To: "Bryan Hewlett" <bhewlett <@t> cogeco.ca>; "Timo Väisänen" 
<timo.vaisanen <@t> oulu.fi>; <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, June 13, 2006 11:27 AM
Subject: RE: [Histonet] Trichrome staining and fibrin


You know full well that MSB required initial staining in mercury. I know 
that zinc is an adequate replacement.
Attempting to get a consistent good red colour with formalin fixed material 
is futile, as is post-fixation.
At times it works and at times not.

IMO, Masson is useless for fibrin.

Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
 Consultant Pathologist
 Rotherham General Hospital
 South Yorkshire
        terry.marshall <@t> rothgen.nhs.uk

-----Original Message-----
From: Bryan Hewlett [mailto:bhewlett <@t> cogeco.ca]
Sent: 13 June 2006 15:52
To: Timo Väisänen; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Trichrome staining and fibrin


Try the Lendrum MSB trichrome variant, it was designed to demonstrate
I will send the method via a separate e-mail.


----- Original Message ----- 
From: "Timo Väisänen" <timo.vaisanen <@t> oulu.fi>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, June 13, 2006 10:33 AM
Subject: [Histonet] Trichrome staining and fibrin

> Hello all,
> I have struggled with getting Masson Trichrome (Goldner) staining to work
> with
> kidney biopsies. The problem is that our pathologist does not get positive
> fibrin staining to glomeruli in diseased kidneys. Those areas of the
> glomeruli
> that should contain fibrin, at least according to the pathologist, are
> green
> (Lichtgrun) and not red, as they should be, whatever I do. I have tried to
> enhance the staining with Bouin's fixative (+56C) pretreatment but it did
> not
> change the overall situation. However, the red stain was more intense in
> skin
> samples containing fibrin (formalin fixed). I have also tried another
> Trichrome
> staining with Crocein Scarlet 7B without any luck. In our lab kidney
> biopsies
> are routinely fixed with alcoholic Bouin's. As far as I know it should be
> compatible with the Masson protocol we use. Any ideas? Any good protocols
> that
> I could try?
> Thank's in advance,
> Timo
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