Re: [Histonet] PAS vs CAS
|From:||Bryan Llewellyn (by way of histonet)|
In my haste I didn't note the 42C temperature. Try it at room temperature.
----- Original Message -----
From: "Bryan Llewellyn"
Sent: Tuesday, October 18, 2005 1:05 PM
Subject: Re: [Histonet] PAS vs CAS
> Try oxidising in the 5% chromic acid for about an hour. That was Bauer's
> original recommendation (4% for 40-60 minutes). Alternatively, many have
> found a 10% solution for 10 minutes satisfactory. You could also do a
> double oxidation with periodic acid. That is oxidise in 1% periodic acid
> for 15 minutes, block aldehyde with aniline-acetic for 30 minutes, then
> re-oxidise for 15 minutes. This also reduces background positive PAS
> Bryan Llewellyn
> Bryan Llewellyn
> ----- Original Message -----
> From: "Cindy DuBois"
> To: "Histonet"
> Sent: Tuesday, October 18, 2005 10:46 AM
> Subject: [Histonet] PAS vs CAS
>> My lab manager recently read the article by Vinnie Della Speranza et. al.
>> on fungus staining. He wanted us to try the Chromic Acid Schiff's (CAS)
>> procedure and compare it to our PAS procedure.
>> Although I get some staining with the CAS, only about half of the fungus
>> is staining and it is very light.
>> I used the following procedure:
>> 1. Clear & Hydrate slides (standard protocol).
>> 2. Incubate slides in 5% Chromic acid in water bath (temp approx. 42C)
>> for 10minutes.
>> Solution was pre-heated for 10 minutes.
>> 3. Rinse and incubate in Schiff's solution for 10 minutes.
>> 4. Rinse in hot water.
>> (Our tap water has a lot of sulfur in it and therefore we do not normally
>> do the sulfuric acid rinse).
>> 5. Counterstain in Working Light Green, dehydrate and coverslip.
>> If any one is using this procedure for their fungus stain would you
>> please send me your procedure?
>> Cindy DuBois, HT ASCP
>> Integrated Pathology Associates
>> Stockton CA
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