Hi Allison!

Without knowing which dyes your kit is employing, I would have to say the lack of an aqueous rinse of some sort is your culprit.  Regardless whether you have an Aniline Blue or Light green collagen stain component, all of the dyes in your typical Gomori one step are far more soluble in water than alcohol.  The Aniline Blue is probably the worst of the batch being barely soluble in alcohol.  We use a 1% Acetic Acid rinse (approx 5 dips) to rinse most of the dyes off before putting it in 95% ETOH then 100% ETOH and up to Xylene. We also have charged slides in our runs and they work fine for us.

If you don't want to stock that extra reagent, just give them a quick swish in dH2O before you run them up thru ETOH.

You will also see this phenomenon in your Retics if you forget to rinse off your Nuclear fast red (found that out the hard way!).

Hope this might help!  Good Luck!

Kathy Johnston
Tech II Special Stains
Anatomic Pathology
Calgary Laboratory Services
#9 - 3535 Research Road NW
Calgary, AB.
403-770-3572


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Scott,
Allison D
Sent: October 25, 2006 10:51 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Gomori's trichrome problem


Hello to all in histoland.  We are currently having a problem that has just cropped up within the last few weeks with our gomori's one step trichrome stain.  It is being done as the procedure says, but when the slides come out of the one step solution and they are rinsed in 100% alcohol, they have a white filmy background.  I think that it has something to do with the sections being on a charged slide.  The only thing that we are not doing according to procedure is not going directly to 0.5% acetic acid solution after the one step solution.   Any help will be appreciated.  JACHO is in the house, I hope that they don't come in my area.
 

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas

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