[Histonet] Trichrome & Fixation

[Amos Brooks]

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
>