AW: [Histonet] Trichrome & Fixation

[Gudrun Lang]

Hi,
years ago we did a stain called CAB (=one step trichrome) regularly on liver-tissue. I don't know if it was because of ignorance or with aim, but it was done without Bouin. The result was blue-grey hepatocytes and darker blue collagen.  - also totally different to the result with Bouin (red hepatocytes).

I think the Bouin is less a "re-fixation" than more an "binding-site retrival" in this context.

Gudrun


-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Amos Brooks
Gesendet: Donnerstag, 05. März 2015 22:01
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Trichrome & Fixation

Hi,
     It is interesting that you should mention the importance of fixation on the Trichrome stain. I have an image of two murine hearts processed, cut and stained side by side. The only difference between the two is that they were harvested at different times, so one sat in formalin long enough to be properly fixed the other one was placed in the fixative then immediately brought in to be processed from 70% ethanol on. They are *totally* different looking. The red muscle tissue looks more purple therefore less distinct from the blue blood vessels. You get a similar effect with lung bronchial epithelium.

Cheers,
Amos

On Tue, Mar 3, 2015 at 1:00 PM, <histonet-request <@t> lists.utsouthwestern.edu>
wrote:

> Message: 15
> Date: Tue, 03 Mar 2015 12:34:33 -0500
> From: John Kiernan <jkiernan <@t> uwo.ca>
> Subject: Re: [Histonet] Masson trichrome and H and E
> To: Emily Brown <talulahgosh <@t> gmail.com>,
>         "histonet <@t> lists.utsouthwestern.edu"
>         <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <7390afa3112a.54f5aa59 <@t> uwo.ca>
> Content-Type: text/plain; CHARSET=US-ASCII
>
> If you can't get two colours with H&E, don't expect to get the colour 
> scheme right with Masson's trichrome, which needs more skill. If you 
> are hoping to show basement membranes in the kidney, you would do 
> better to use a technically simpler staining method such as 
> picro-sirius red or periodic acid-Schiff. If for some reason you 
> really need three colours, a one-step trichrome such as Gomori's, 
> Cason's or Gabe's might be the way to go rather than Masson's or one 
> of the other multi-step trichromes. Remember that all trichrome 
> methods are greatly influenced by the fixative. A post-fixation 
> treatment of the sections, usually with picric acid, is needed for 
> formaldehyde-fixed tissues. Some alternative post-fixation treatments 
> were proposed by Yu & Chapman (2003) J. Histotechnol. 26(2): 131-134, but their coloured photos were not very convincing.
>
> Making up staining solutions in-house is always cheaper than buying 
> pre-made solutions.
>
> John Kiernan
> London, Canada
>
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