Re: [Histonet] PAS Stain

2018-11-05 Thread Mayer,Toysha N via Histonet 
Laurie,

The warm water helps the color of the Schiff develop in a more vibrant pattern. 
 If you allow the slides to sit in warm water for 2 min, then rinse you also do 
not have to use the metabisulfite rinses to remove excess stain.

Toysha N. Mayer D.H.Sc., MBA, HT(ASCP)
Assistant Professor/Education Coordinator
HTL Program
MD Anderson School of Health Professions
713.563.3481
tnma...@mdanderson.org









--

Message: 3
Date: Mon, 5 Nov 2018 16:44:23 + (UTC)
From: Laurie Redmond 
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS Stain
Message-ID: <1715295197.1084926.1541436263...@mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

Does anyone else out there use warm water to rinse the PAS slides after the 
Schiff's reagent?? Can anyone comment on whether using warm water vs. cold 
water would have any affect on the stain?
Laurie Redmond HT (ASCP)

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Re: [Histonet] PAS Stain

2016-05-09 Thread Victor via Histonet 

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Re: [Histonet] PAS Stain

2016-05-09 Thread Seeley, Heather via Histonet 
We have one girl that always spits on our slide! :) works great!

HEATHER SEELEY, HT(ASCP)
Histotech
803-985-4676 OFFICE
803-327-7598 FAX



From: Bob Richmond [rsrichm...@gmail.com]
Sent: Thursday, May 05, 2016 2:10 PM
To: Histonet@lists.utsouthwestern.edu
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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Re: [Histonet] PAS Stain

2016-05-06 Thread Rene J Buesa via Histonet 
As I see it, there are 3 main objections about using human saliva as an amylase 
source.In order of importance they are:1- you will never know the actual 
concentration of the amylase and this will produce reproducibility problems.2- 
along with the saliva you will introduce bacteria that may end being stained 
and can cause misdiagnoses.3- it is absolutely disgusting.René 

On Thursday, May 5, 2016 2:26 PM, Bob Richmond via Histonet 
 wrote:
 

 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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Re: [Histonet] PAS Stain

2016-05-05 Thread Ray via Histonet 
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 t

Re: [Histonet] PAS Stain

2016-05-05 Thread Sherry Martin via Histonet 
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
>>
>>
>>

Re: [Histonet] PAS Stain

2016-05-05 Thread Ingles Claire via Histonet 
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
&g

Re: [Histonet] PAS Stain

2016-05-05 Thread Tony Henwood (SCHN) via Histonet 
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
>>
>>
> ___
> Histonet mailing list

Re: [Histonet] PAS Stain

2016-05-05 Thread Anne Murvosh via Histonet 
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
>>
>>
> ___
> Histonet mailing list
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>
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-- 
--
**
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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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Re: [Histonet] PAS Stain

2016-05-05 Thread Ingles Claire via Histonet 
The level of amylase in your saliva also depends on when you ate last. If 
you're spitting on slides after you just ate, the reaction will be weaker as 
the amylase will have been used up on lunch digestion. (also, you need to be 
careful about having your lunch salad contaminate the slide... :)
Claire


From: Ray via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Thursday, May 05, 2016 1:48 PM
To: Richmond, Bob
Cc: Histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] PAS Stain

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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Re: [Histonet] PAS Stain

2016-05-05 Thread Geoff via Histonet 
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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Re: [Histonet] PAS Stain

2016-05-05 Thread Anne Murvosh via Histonet 
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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Re: [Histonet] PAS Stain

2016-05-05 Thread Ray via Histonet 
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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Re: [Histonet] PAS Stain

2016-05-05 Thread Ray via Histonet 
Hello 
  
Agree with comments about temperature of 60 degrees killing an enzyme.  If you 
plot enzyme activity on "y"  axis and temperature on "x" axis it is not a 
straight line.  Every enzyme has an optimal temperature and can function slowly 
at non-optimal or optimally at correct temperature.  Diastase/amylase from your 
body work at 37 degrees, less so or dead at RT or 60 degrees.  Nuking a mixture 
to 37 degrees might nuke protein.  Nuke buffer to 37, verify temp and add the 
enzyme. 
  
pH-if you plot enzyme activity on "y" axis and pH on x-axis, it is not a 
straight line.  There is an optimal pH for each enzyme.  The enzyme might work 
fine at different pH; just not nearly as well. 
  
I know some still use 0.5% or 1.0% in di water.  I'd advise not to.  Use a 
buffer at a given pH.  "pHíng" di or distilled water with ordinary lab 
electrodes is an act of futility.  From a standard lab electrode point of view, 
there is NO pH to read.  Not enough ions present so meter might say anything.  
If anything, it will pick up CO2 from air which becomes carbonic acid and meter 
will flail wildly in acidic region.  But that is not a stable buffer.  di water 
might work.  Just that sometimes, it might not.  Why chance it?  di water has 
unstable (no reliably readable) pH. 
  
Ray Koelling, golfing in Spokane WA and enjoying the low humidity 

- Original Message -

From: "Joanne Clark via Histonet"  
To: histonet@lists.utsouthwestern.edu 
Sent: Wednesday, May 4, 2016 1:02:48 PM 
Subject: [Histonet] PAS Stain 

Hi Histonetters, I'm hoping someone can help me troubleshoot our PAS diastase 
method.  We have been digesting the tissue in 0.5% diastase of malt in a 60 
degree oven for 30 minutes, but do not see the glycogen being digested out.  I 
have tried alpha amylase and beta amylase also without any luck.  Does anyone 
have any suggestions to get the digestion to work 

Joanne Clark, BAAS, HT(ASCP)CM 
Director of Histology 

P.   (575) 622-5600 
C.   (575) 317-6403 
F.   (575) 622-3720 
TF. (800) 753-7284 

pcnm.com 




Disclaimer: This electronic message may contain information that is 
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recipient of this message, please notify the sender immediately and delete the 
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Re: [Histonet] PAS Stain

2016-05-05 Thread Jennifer MacDonald via Histonet 
We switched form malt diastase to alpha amylase and have seen a 
significant improvement in digestion, but as others have said enzyme 
activity will be destroyed at 60 degrees C. We put ours into a 37 degree C 
water bath.  Enzymes work optimally at 37C, body temperature.  As the 
temperature rises above 37C you will see a decrease in enzyme activity and 
then none at all.
Jennifer



From:   Joanne Clark via Histonet 
To: "histonet@lists.utsouthwestern.edu" 

Date:   05/04/2016 01:04 PM
Subject:[Histonet] PAS Stain



Hi Histonetters, I'm hoping someone can help me troubleshoot our PAS 
diastase method.  We have been digesting the tissue in 0.5% diastase of 
malt in a 60 degree oven for 30 minutes, but do not see the glycogen being 
digested out.  I have tried alpha amylase and beta amylase also without 
any luck.  Does anyone have any suggestions to get the digestion to work

Joanne Clark, BAAS, HT(ASCP)CM
Director of Histology

P.   (575) 622-5600
C.   (575) 317-6403
F.   (575) 622-3720
TF. (800) 753-7284

pcnm.com




Disclaimer: This electronic message may contain information that is 
proprietary,
confidential, or legally privileged or protected. It is intended only for 
the use
of the individual(s) and entity named in the message. If you are not an 
intended
recipient of this message, please notify the sender immediately and delete 
the
material from your computer. Do not deliver, distribute or copy this 
message and
do not disclose its contents or take any action in reliance on the 
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contains.
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Re: [Histonet] PAS Stain

2016-05-05 Thread Bob Richmond via Histonet 
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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Re: [Histonet] PAS Stain

2016-05-05 Thread Ray via Histonet 
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 

- Original Message -

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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Re: [Histonet] PAS Stain

2016-05-05 Thread Bob Richmond via Histonet 
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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Re: [Histonet] PAS Stain

2016-05-05 Thread Manfre, Philip via Histonet 
We switched to alpha amylase since the diastase we were ordering was not 
working.  We use a 2% solution of alpha amylase for 40 minutes at room 
temperature and it has been working fine.

Phil.

Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_man...@merck.com



-Original Message-
From: Joanne Clark via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, May 04, 2016 4:03 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS Stain

Hi Histonetters, I'm hoping someone can help me troubleshoot our PAS diastase 
method.  We have been digesting the tissue in 0.5% diastase of malt in a 60 
degree oven for 30 minutes, but do not see the glycogen being digested out.  I 
have tried alpha amylase and beta amylase also without any luck.  Does anyone 
have any suggestions to get the digestion to work

Joanne Clark, BAAS, HT(ASCP)CM
Director of Histology

P.   (575) 622-5600
C.   (575) 317-6403
F.   (575) 622-3720
TF. (800) 753-7284

pcnm.com




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Re: [Histonet] PAS Stain

2016-05-05 Thread Gudrun Lang via Histonet 
As far as I remember the incubation temperature is at roomtemperature. 60°C
would rather denature the native enzyme than increase activity.

Look at the optimal working temp of the reagens you have bought.

regards
Gudrun

-Ursprüngliche Nachricht-
Von: Joanne Clark via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 04. Mai 2016 22:03
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] PAS Stain

Hi Histonetters, I'm hoping someone can help me troubleshoot our PAS
diastase method.  We have been digesting the tissue in 0.5% diastase of malt
in a 60 degree oven for 30 minutes, but do not see the glycogen being
digested out.  I have tried alpha amylase and beta amylase also without any
luck.  Does anyone have any suggestions to get the digestion to work

Joanne Clark, BAAS, HT(ASCP)CM
Director of Histology

P.   (575) 622-5600
C.   (575) 317-6403
F.   (575) 622-3720
TF. (800) 753-7284

pcnm.com




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Re: [Histonet] PAS Stain

2015-06-03 Thread Mayer,Toysha N 
We use the hematoxylin that is used on the HE stainer.  While I know it is not 
optimal, the pathologists should be  used to looking at it so it makes things a 
lot easier.  The time is cut down to less than 30 seconds and we do not use a 
differentiator.
For the students, it helps them to understand the versatility of the dye and 
its multiple uses.  They don't get confused and are pleasantly surprised when 
they find out they can use another formulation while they are at their 
internships.  

Sincerely,

Toysha N. Mayer, D.H.Sc., MBA, HT (ASCP)
Instructor/Education Coordinator
Program in Histotechnology
School of Health Professions
UT M.D. Anderson Cancer Center
713.563-3481




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Message: 2
Date: Mon, 1 Jun 2015 22:02:55 -0500
From: Mica faith14...@aol.com
To: histonet@lists.utsouthwestern.edu
histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS stain
Message-ID: 16f1d792-cd79-458e-9d25-493d8bc0c...@aol.com
Content-Type: text/plain; charset=us-ascii

Just a question...what kind of hematoxylin do you use as a counter stain for 
your PAS stain and do you use a differentiator?

Sent from my iPad


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Re: [Histonet] PAS stain

2015-06-02 Thread Tony Henwood (SCHN) 
Hi Mica,

Most haematoxylins will do eg Harris or Mayer's.
I would recommend a light counterstain (it is a counterstain, not the main 
stain).
Differentiate if too strong.

Also periodic acid treatment increases nuclear affinity for Hx so shorten the 
counterstain time.

Regards,
Tony

From: Mica [faith14...@aol.com]
Sent: Tuesday, 2 June 2015 1:02 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS stain

Just a question...what kind of hematoxylin do you use as a counter stain for 
your PAS stain and do you use a differentiator?

Sent from my iPad
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RE: [Histonet] PAS STAIN

2010-10-06 Thread Rathborne, Toni 
Were the new bottles from the same lot? 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Diana
McCaig
Sent: Wednesday, October 06, 2010 11:03 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS STAIN


Hi
We have been doing a PAS stain on the same control block and same
reagent supplier for a long time.  
Yesterday when we ran the slides, they failed to stain
 
We opened new bottles of Periodic Acid and Schiff's Reagent and recut
fresh controls
 
Still, we are unable to get any staining.
 
Suggestions to help would be appreciated
 
Diana
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RE: [Histonet] PAS STAIN

2010-10-06 Thread gayle callis 
I suspect bad Periodic acid.  We never buy periodic acid already in solution 
and if that has been sitting around even in a kit, it may have gone bad.  In a 
workshop taught by Charles Culling, an expert on PAS staining, he stressed 
making periodic acid fresh daily or each time the stain is done.  It is not 
expensive, goes into solution readily, then toss the periodic acid to never be 
reused with exception of that one day. Also, you can test your Schiffs reagent 
by putting a few drops of Schiffs into 10 mls NBF, watch it turn a very bright 
pink red instantly.   If it turns purplish, it is not good. It may be a bad 
kit, but you never said you were using a kit only new bottles of .

Consequently we buy periodic acid in crystalline form, make up 1% Periodic 
acid, and buy the Schiffs Reagent from Fisher, never a kit.  Sigma also sells 
good Schiffs, but Fisher Scientific aka Thermo has larger quantity for less 
price.  We also store our Schiffs in the refrigerator rather than room 
temperature, a habit left over from days when we made this reagent up in house. 
 

Gayle M. Callis 
HTL/HT/MT(ASCP)  



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni
Sent: Wednesday, October 06, 2010 9:06 AM
To: Diana McCaig; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] PAS STAIN

Were the new bottles from the same lot? 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Diana
McCaig
Sent: Wednesday, October 06, 2010 11:03 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] PAS STAIN


Hi
We have been doing a PAS stain on the same control block and same
reagent supplier for a long time.  
Yesterday when we ran the slides, they failed to stain
 
We opened new bottles of Periodic Acid and Schiff's Reagent and recut
fresh controls
 
Still, we are unable to get any staining.
 
Suggestions to help would be appreciated
 
Diana
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printing, copying, distribution, or use of such information is strictly
prohibited and may be unlawful.  If you are not the addressee, please
promptly delete this message and notify the sender of the delivery error
by e-mail or you may call Somerset Medical Center's computer Help Desk
at 908-685-2200, ext. 4050.

Be sure to visit Somerset Medical Center's Web site - 
www.somersetmedicalcenter.com - for the most up-to-date news, 
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Re: [Histonet] PAS STAIN

2010-10-06 Thread Geoff McAuliffe 

I make my Schiff's from scratch and store it in the refrigerator.
I use a minimum amount of charcoal and I heat it in the oven at over 100 
C the night before to be sure it is nice and dry.

I make periodic acid fresh that day.
I make bisulfite rinses fresh that day.
Aqueous formalin for 48 hours at room temp. is OK for rat and mouse 
liver glycogen, I don't know about other species. Slices of liver should 
be thin.

When in doubt formalin+alcohol+acetic acid is an excellent fixative.

Geoff


Diana McCaig wrote:

Hi
We have been doing a PAS stain on the same control block and same
reagent supplier for a long time.  
Yesterday when we ran the slides, they failed to stain
 
We opened new bottles of Periodic Acid and Schiff's Reagent and recut

fresh controls
 
Still, we are unable to get any staining.
 
Suggestions to help would be appreciated
 
Diana

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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 
mcaul...@umdnj.edu

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RE: [Histonet] PAS STAIN

2010-10-06 Thread sgoebel 

   I  know  there were a ton of posts about this, but I deleted them o= n
   accident...Did  you test your Schiff's?  Put a drop of 10% formalin i   n 
it...if it turns pink, it's good...if not, throw it out =)

   Sarah Goebel= , B.A., HT (ASCP)

   Histotechnician

   XBiotech US= A Inc.

   8201 East Riverside Dr. Bldg 4 Suite 100
   A= ustin, Texas  78744
   (512)386-5107

    Original Message 
   Subject: Re: [Histonet] PAS STAIN
   From: Geoff McAuliffe [1]mcaul...@um= dnj.edu
   Date: Wed, October 06, 2010 1:28 pm
   To: Diana McCaig [2]dmcc...@ckha.on.= ca, Histonet ((E-mail))
   [3]histo...@lists.uts= outhwestern.edu
   I make my Schiff's from scratch and store it in the refrigerator.
   I  use  a minimum amount of charcoal and I heat it in the oven at over
   100 C the night before to be sure it is nice and dry.
   I make periodic acid fresh that day.
   I make bisulfite rinses fresh that day.
   Aqueous formalin for 48 hours at room temp. is OK for rat and mouse
   liver  glycogen,  I  don't  know  about other species. Slices of liver
   should be thin.
   When in doubt formalin+alcohol+acetic acid is an excellent fixative.
   Geoff
   Diana McCaig wrote:
Hi
We have been doing a PAS stain on the same control block and same
reagent supplier for a long time.
Yesterday when we ran the slides, they failed to stain
   
 We  opened  new  bottles  of Periodic Acid and Schiff's Reagent and
   recut= br  fresh controls
   
Still, we are unable to get any staining.
   
Suggestions to help would be appreciated
   
Diana
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[5]= 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
   [6]mcaul...@umdnj.edu
   **
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References

   1. 3Dmailto:mcaul...@umdnj.edu;
   2. 3Dmailto:dmcc...@ckha.on.ca;
   3. 3Dmailto:histonet@lists.utsouthwestern.edu;
   4. 3Dmailto:Histonet@lists.utsouthwestern.edu;
   5. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
   6. 3Dmailto:mcaul...@umdnj.edu;
   7. 3Dmailto:Histonet@lists.utsouthwestern.edu;
   8. 3Dhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet;
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