Indeed, a very curious and interesting way to do this.  And as I said-have done 
it very occasionally in long ago past. 
What I am still curious about is that for those who still do this, how do you 
write it up for CLIA or CAP or GLP when, as the Samuri Pathologist would call 
them, Herrn Inspektors come to visit.  Lot number of spit?  Variation of spit 
reagent with a different lot (person)?  Preparation of spit reagent (before or 
after starchy meal)?  What the step-by-step procedure looks like?  Just 
curious. 
  
Spokane Ray 

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

From: "Sherry Martin via Histonet" <histonet@lists.utsouthwestern.edu> 
To: histonet@lists.utsouthwestern.edu 
Sent: Thursday, May 5, 2016 5:30:22 PM 
Subject: Re: [Histonet] PAS Stain 

Hello All! I've served in laboratory medicine for well over 35 years and in my 
early years we did indeed spit on the slides. I learned very quickly on that my 
personal spit fails to digest any glycogen. And worse yet, I used to have to 
search for someone else to spit for me. PAS stains were always awkward and 
gross! I'm VERY thankful for the artificial amylase we have now. :-) 
Y'all have a great day! 
Sherry Martin 

    On Thursday, May 5, 2016 7:06 PM, Ingles Claire via Histonet 
<histonet@lists.utsouthwestern.edu> wrote: 
  

 I don't know, I believe Dr. Salk did the same with the polio vaccine. He even 
involved his family! Dedicated doctors... 
Claire 

________________________________________ 
From: Tony Henwood (SCHN) via Histonet [histonet@lists.utsouthwestern.edu] 
Sent: Thursday, May 05, 2016 6:33 PM 
To: Anne Murvosh 
Cc: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] PAS Stain 

Only a crazy Aussie would do this!! 

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead 
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department 
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message----- 
From: Anne Murvosh via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, 6 May 2016 6:03 AM 
To: Histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] PAS Stain 

You clearly don't know your histo history. The reason we know that H pylori 
exists is because a Scientist, Dr. Barry Marshall wanted to prove bacteria 
caused ulcers and not stress.  No one believed him.  So he took the organisms 
from a patient, mixed it in a broth and drank it. He then biopsied himself and 
treated it. There's a non-uniform method that saved a lot of suffering.  Bravo 
we crazy scientists.  Anne 


-----Original Message----- 
From: Geoff via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, May 05, 2016 12:31 PM 
To: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] PAS Stain 

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 
>> _______________________________________________ 
>> Histonet mailing list 
>> Histonet@lists.utsouthwestern.edu 
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>> 
>> 
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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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