[Histonet] PAS Stain

[Geoff]

I cannot believe any scientist would advocate such a non-uniform method 
as spitting on a slide.
Buy a bottle of what ever enzyme and use a reproducible buffer and 
temperature.

Geoff

On 5/5/2016 3:19 PM, Anne Murvosh via Histonet wrote:
> Yes, spitting is the tried and true way to do it.  Not to mention no measuring and cheaper.  The reason we switched to a powder is because I just don't spit well I used to have someone do it for me cause I would end up drooling. YUCK! The best way to find out is do the amylase method and the spit method at the same time and have the doctor pick the best.  A fun experiment  Anne	
>
> -----Original Message-----
> From: Bob Richmond via Histonet [mailto:histonet at lists.utsouthwestern.edu]
> Sent: Thursday, May 05, 2016 11:36 AM
> To: koellingr at comcast.net
> Cc: Histonet at lists.utsouthwestern.edu
> 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, <koellingr at 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 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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>>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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