[Histonet] PAS Stain

[koellingr at comcast.net]

I love having the Samuri Pathologist on this forum for wisdom and real-laboratory life knowledge.  And yes, I have in the past spit on slide ON OCCASSION when faced with a dire necessity.  Although I know there are those who would wretch about this; it remains a fact of viable laboratory life for some. 
  
My problem now is that in this era of (MUCH TOO MUCH) regulation, how do you "test lots" or control from "lot-to-lot variation" in this SOP?  When Jane or Joe do this routinely and then goes on vacation, what about Sally or Jim spit?  There is a variation in copy number of the AMY1 gene (amylase) and resulting difference in amylase protein concentration amongst individuals. 
  
Why not just standardize it from the start, reagent, pH, temperature and it really cannot fail. 
  
Spokane Ray 

----- Original Message -----

From: "Bob Richmond via Histonet" <histonet at lists.utsouthwestern.edu> 
To: "Histonet at lists.utsouthwestern.edu" <histonet at lists.utsouthwestern.edu> 
Sent: Thursday, May 5, 2016 11:10:40 AM 
Subject: Re: [Histonet] PAS Stain 

Amylase (diastase) for the PAS stain queries: 

Whatever happened to spitting on the slide (30 min at room temperature)? 
John Kiernan advises "thinking of lemons and drooling into a small beaker" 
though I'd advise chewing on a rubber band for a few seconds. 

He notes that alpha amylase is preferred. I'd go with the cheapest one in 
the Sigma-Aldrich catalog. Room temperature is usual, but I note that Sigma 
offers a heat-stable alpha amylase. 

Bob Richmond 
Samurai Pathologist 
Maryville TN 
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