An excellent point.  For anyone wanting to investigate-simply do a PubMed 
search on variation of AMY1 gene.  Sorry; I guess I should say this is, 
strictly speaking, non-histology related topic and I don't want to get into 
trouble as some before me.  Tons of research about this linking back (in 
theory) to positive selection in hunter-gatherers versus agricultural 
ancestors, race, obesity, phenotypic and dietary differences as to why maybe 
there can be big differences. 
Spokane Ray 

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

From: "Bob Richmond" <rsrichm...@gmail.com> 
To: koelli...@comcast.net 
Cc: "Histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> 
Sent: Thursday, May 5, 2016 11:35:42 AM 
Subject: Re: [Histonet] PAS Stain 

Spokane Ray points out something I've wondered about for years - can just 
anybody spit on the slide and remove the glycogen? I've never heard of any 
variation, but the number of people I've asked is very limited. This reference: 
http://www.ncbi.nlm.nih.gov/gene/276 
certainly suggests that different people have different salivary alpha amylase 
activity. 

Bob Richmond 

On Thu, May 5, 2016 at 2:27 PM, < koelli...@comcast.net > wrote: 



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 


From: "Bob Richmond via Histonet" < histonet@lists.utsouthwestern.edu > 
To: " Histonet@lists.utsouthwestern.edu " < histonet@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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