[Histonet] Trichrome troubleshooting

[Michael Ann Jones]

Don¹t know if this matters, but we have found that the trichrome stain
fades on tissues that are not cut fresh.
We use uterus as a control but they must be cut fresh, no batching this
one. 
The trichrome is not as brilliant on pre-cut tissues.
Just my 2 cents :)

Michael Ann
Michael Ann Jones, HT (ASCP)
Histology Manager
Metropath
7444 W. Alaska Dr. #250
Lakewood, CO 80226
303.634.2511
Mjones at metropath.com






On 6/30/15, 2:29 PM, "Elizabeth Chlipala" <liz at premierlab.com> wrote:

>Suzanne
>
>How many times have you used the kit and reagents, I did look up how the
>kit works but the trichrome stain can be tricky.  First of all you need
>to make sure that the mordant (bouins solution) is at 60C prior to
>placing your slides in them.  We normally heat up our bouins for at least
>an hour prior to placing the slides in the solution.  We leave in bouins
>for an hour and a half rather than an hour.   I see that this is a
>microwave protocol I cannot comment on that but I don't think that the
>hematoxylin is the issue, if you leave longer in 1% acetic acid that may
>pull some of the blue stain out or I would try dehydrating with lower
>alcohol percentage that can pull some of the blue stain out.   I would
>also try leaving it a bit longer in the bouins after you microwave it -
>that might help.
>
>Trichrome staining works best with fresh reagents so if you have used
>these reagents too much that could cause problems.  I'm also not a big
>fan of the one step trichromes, they are quicker but sometimes not as
>good as the two steps, just my opinion.
>
>FYI - to evaluate your staining look for a smaller vessel, the smooth
>muscle should be nice a red, if its greyish or blue you have not done the
>stain properly.  Good Luck
>
>Liz
>
>Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
>Premier Laboratory, LLC
>PO Box 18592
>Boulder, CO 80308
>(303) 682-3949 office
>(303) 682-9060 fax
>(303) 881-0763 cell
>liz at premierlab.com
>www.premierlab.com
>
>March 10, 2014 is Histotechnology Professionals Day
>
>Ship to Address:
>
>Premier Laboratory, LLC
>1567 Skyway Drive, Unit E
>Longmont, CO 80504
>
>
>-----Original Message-----
>From: Suzanne Martin [mailto:smartin at lcpath.com]
>Sent: Tuesday, June 30, 2015 12:37 PM
>To: histonet at lists.utsouthwestern.edu
>Subject: [Histonet] Trichrome troubleshooting
>
>
>Hi all,
>
>We are having trouble troubleshooting our trichrome. It is too blue. We
>are using Leica's kit with the Weigerts iron with Gills. Most of the
>small bowel controls have seen improvement but patient tissue is not...
>strange. 
>
>We have tried lessening the time in Gills, adding time for the last acid
>step, even lessening time and adding time in the Weigerts.
>
>
>Thoughts?
>
>Thank you.
>
>Suzanne HT
>
>
>
>
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