[Histonet] PAS Stain

Thu May 5 20:54:37 CDT 2016

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 at lists.utsouthwestern.edu> 
To: histonet at 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 at 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 at lists.utsouthwestern.edu] 
Sent: Thursday, May 05, 2016 6:33 PM 
To: Anne Murvosh 
Cc: histonet at 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 at lists.utsouthwestern.edu] 
Sent: Friday, 6 May 2016 6:03 AM 
To: Histonet at 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 at lists.utsouthwestern.edu] 
Sent: Thursday, May 05, 2016 12:31 PM 
To: histonet at 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 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 
>> _______________________________________________ 
>> Histonet mailing list 
>> Histonet at lists.utsouthwestern.edu 
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>> 
>> 
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Neuroscience and Cell Biology 
Robert Wood Johnson Medical School 
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