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@lists.utsouthwestern.edu]
Sent: Thursday, May 05, 2016 11:36 AM
To: koelli...@comcast.net
Cc: Histonet@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, <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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**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732) 235-4583; fax: -4029
mcaul...@rwjms.rutgers.edu
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