Re: PAS Problems, more info Gayle
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From: | Jerry Wilson <jwilso70@bellsouth.net> |
To: | Donna Carr <dkc@odsgc.net> |
Reply-To: | |
Date: | Wed, 07 Apr 1999 21:23:14 +0000 |
Content-Type: | text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" |
Donna-
I've always been under the impression that the Schiffs should be
refrigerated, whether obtained from commercial sources or prepared in
house. The periodic acid could be stored at room temp. However, i learned
these techniques several years ago and maybe some things have changed.
They still work for me..might be worth a try..
good luck,
jerry
nola
Donna Carr wrote:
> Gayle asked for more info.
> To further elaborate on the problems we are experiencing with our
> PAS stain, I have worked in Histology for 2 years and never had any
> problem with a PAS stain. We also use the spit method for diastase.
> The stain has always worked beautifully. Usually the second the slides
> are placed in the coplin jar the control turns pink. That's why I am so
> baffled at the recent problems. The sections are not staining enough.
> They are turning pink but not reacting like they normally do. We stain
> liver needle biopsies and tumors with our PAS. We used to reuse our
> reagents but have gone to using a staining rack instead of coplin
> jars. I did try the coplin jar method without any success.
>
> Our method is as follows:
> hydrate to di H20
> 5 min periodic acid (1g/dl)
> rinse in DI
> 15 min in Schiff's reagent
> 2 min rinse in Sodium metabisulfate solution
> 2 min rinse in Sodium metabisulfate solution
> 5 to 10 min rinse in running tap water to enhance pink
> stain 4 min Hematoxylin
> dehydrate, clear, cover slip
> this is from memory and I can't remember the concentration of the
> Sodium metabisulfate? (0.5g/100ml)
>
> 1. We used a different control, so this was the first suspected
> culprit. We obtained new sections from autopsy liver for controls and
> ran several trials with different old control blocks and new blocks.
> Not seeing any improvement in the staining.
> 2. We had problems with our reagents having ice crystals in them (
> we share a fridge with chemistry) and have since moved our reagents to a
> different location. I called both companies that we get our Periodic
> acid and Schiff's reagent from and both assured me that freezing would
> NOT harm to the reagents. Only to make sure that the Periodic acid was
> thoroughly mixed and at room temp before using. Concerning the Schiff's
> reagent I was told that it was okay to keep at room temp and no need to
> refrigerate.
> 3. We are a small lab and keep reagents as long as possible.
>
> My plan of action:
> We have a new bottle of Schiff's that we are now using. So I think the
> first thing to do is to get a section of cervix to use as a control.
> Secondly to use a new batch of periodic acid. I don't believe we have
> the chemicals to make our periodic acid, but have a small amount used
> for our fungus stain that is the same concentration.
>
> Any further suggestions are welcome, sorry so long winded. Thanks again
> for all the response.
> Sincerely Donna
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