[Histonet] Trichrome

[Amos Brooks]

Hi,
      It is imperative to make sure the fixation is started immediately and
the tissue is fully fixed. Trichrome stains are *notoriously* affected by
fixation. I have an image of two lung sections processed simultaneously,
embedded and cut at the same time and stained at the same time. The
epithelium of one was a nice pink and the other had faded to purple and
almost blue. The only difference between them was the duration of fixation.
If your control tissue is working and the test tissue is not both on the
autostained and manually, I would strongly suspect the fixation of the test
tissue. One final test could be to make up fresh solutions and try again,
although I would suspect you are likely to get more of the same.

All the best,
Amos Brooks

On Tue, Dec 4, 2012 at 1:00 PM,
<histonet-request <@t> lists.utsouthwestern.edu>wrote:

> Message: 3
> Date: Tue, 4 Dec 2012 05:18:35 -0800 (PST)
> From: Cheryl <tkngflght <@t> yahoo.com>
> Subject: [Histonet] Trichrome
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>         <1354627115.8370.YahooMailClassic <@t> web39405.mail.mud.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hi- need help troubleshooting a trichrome.  The stain on the Nexus
> stainers is inconsistent.  Taking it off the stainer and doing it by hand
> with a kit has similar results.
>
> The tissue is NBF fixed needle biopsies on kidney.  There is a little
> mechanical damage on the edges of the tissue but the collagens are staining
> too light and the red overstains.  The normal tissue as control is
> beautiful -- I know I know--don't force the stain--but it doesn't make
> sense to have this much variation from a kit or auto stainer...
>
> Help?
>
>