Re: [Histonet] Alcian blue van gieson double stain

[Tony Reilly via Histonet]

Hi Charles

I have done this stain many times as an abbreviated version of Movats 
Pentachrome.
You are correct in thinking the van gieson will affect the AB stain due to the 
acid.

As in the Movat stain do AB first then place slides in alcoholic ammonia for 1 
hour to convert AB to insoluble Monastral blue then finish with the VVG.

Regards
Tony Reilly

Sent from my iPhone

> On 10 Mar 2023, at 5:49 am, Akemi Allison via Histonet 
>  wrote:
> 
> Just curious, why not do a Movat’s Pentachome? It gives you everything you 
> are looking for and more.  
> Best regards,
> Akemi Allison-Tacha, BS, Ht, HTL 
> akemiat3...@gmail.com
> 
> Sent from my iPhone
> 
>> On Mar 9, 2023, at 11:19 AM, Paula Keene Pierce via Histonet 
>>  wrote:
>> 
>> Hi,
>> if they are wanting to stain elastic, Alcian blue, and Van Gieson, I 
>> recommend doing the Verhoff elastic stain first, then the Alcian blue, and 
>> then the Van Gieson.
>> Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
>> Blue Lake DriveNorman, OK 73069PH 
>> 405-759-3953http://www.excaliburpathology.com
>> 
>> A sharp knife is nothing without a sharp eye. - Klingon Proverb 
>> 
>>   On Thursday, March 9, 2023 at 01:05:14 PM CST, Charles Riley via Histonet 
>>  wrote:  
>> 
>> I am working with a researcher and they saw an article where someone
>> performed an Alcian Blue  and elastic van Gieson double stain.  If anyone
>> has done this before can you provide your protocol.  I am trying to figure
>> out the best way to perform this process to get the best staining results.
>> Here are my questions/thoughts
>> 
>> 1. Is it better to run the Alcian blue as usual and then perform the van
>> gieson stain?
>> 2. Will the differentiating solution in the van gieson stain affect the
>> alcian blue staining in any way?
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Re: [Histonet] Alcian blue van gieson double stain

[Akemi Allison via Histonet]

Just curious, why not do a Movat’s Pentachome? It gives you everything you are 
looking for and more.  
Best regards,
Akemi Allison-Tacha, BS, Ht, HTL 
akemiat3...@gmail.com

Sent from my iPhone

> On Mar 9, 2023, at 11:19 AM, Paula Keene Pierce via Histonet 
>  wrote:
> 
> Hi,
> if they are wanting to stain elastic, Alcian blue, and Van Gieson, I 
> recommend doing the Verhoff elastic stain first, then the Alcian blue, and 
> then the Van Gieson.
> Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
> Blue Lake DriveNorman, OK 73069PH 
> 405-759-3953http://www.excaliburpathology.com
> 
> A sharp knife is nothing without a sharp eye. - Klingon Proverb 
> 
>On Thursday, March 9, 2023 at 01:05:14 PM CST, Charles Riley via Histonet 
>  wrote:  
> 
> I am working with a researcher and they saw an article where someone
> performed an Alcian Blue  and elastic van Gieson double stain.  If anyone
> has done this before can you provide your protocol.  I am trying to figure
> out the best way to perform this process to get the best staining results.
> Here are my questions/thoughts
> 
> 1. Is it better to run the Alcian blue as usual and then perform the van
> gieson stain?
> 2. Will the differentiating solution in the van gieson stain affect the
> alcian blue staining in any way?
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Re: [Histonet] Alcian blue van gieson double stain

[Paula Keene Pierce via Histonet]

Hi,
if they are wanting to stain elastic, Alcian blue, and Van Gieson, I recommend 
doing the Verhoff elastic stain first, then the Alcian blue, and then the Van 
Gieson.
Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
Blue Lake DriveNorman, OK 73069PH 405-759-3953http://www.excaliburpathology.com

A sharp knife is nothing without a sharp eye. - Klingon Proverb 

On Thursday, March 9, 2023 at 01:05:14 PM CST, Charles Riley via Histonet 
 wrote:  
 
 I am working with a researcher and they saw an article where someone
performed an Alcian Blue  and elastic van Gieson double stain.  If anyone
has done this before can you provide your protocol.  I am trying to figure
out the best way to perform this process to get the best staining results.
Here are my questions/thoughts

1. Is it better to run the Alcian blue as usual and then perform the van
gieson stain?
2. Will the differentiating solution in the van gieson stain affect the
alcian blue staining in any way?
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[Histonet] Alcian blue van gieson double stain

[Charles Riley via Histonet]

I am working with a researcher and they saw an article where someone
performed an Alcian Blue  and elastic van Gieson double stain.  If anyone
has done this before can you provide your protocol.  I am trying to figure
out the best way to perform this process to get the best staining results.
Here are my questions/thoughts

1. Is it better to run the Alcian blue as usual and then perform the van
gieson stain?
2. Will the differentiating solution in the van gieson stain affect the
alcian blue staining in any way?
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Re: [Histonet] Alcian Blue staining

[John Kiernan via Histonet]

Pink with alcian blue?  Are you using an alcian blue pH2.5-PAS sequence?  If so 
you should get PAS-positive mucus (not in goblet cells) in the stomach and 
AB-positive mucus (greenish blue) in the intestine. Intestinal mucus, 
especially in the duodenum and jejunum, is also PAS +ve, so the cells appear 
purplish.  In the colon the mucus is mostly sulphated rather than sialylated, 
so it is only AB +ve, even if the stain is done at pH 1.

If a PAS stain is done before the AB, PAS +ve mucus also stains quite strongly 
with AB. For more about this, see Johannes ML & Klessen C (1984) Alcianblue/PAS 
or PAS/alcianblue. Remarks on a classical technic used in carbohydrate 
histochemistry. Histochemistry 80: 129-132. (Unfortunately the paper has only 
black & white photos.)

Various dyes are currently sold as "alcian blue" and not all are suitable for 
all applications. The Biological Stain Commission now certifies alcian blue 
dyes as either "alcian blue 8G or equivalent" or as "alcian blue variant".  See 
the recently updated (2020) entry for Alcian blue 8G (CI 74240) and other 
alcian blue dyes at   https://biologicalstaincommission.org/new-dyes/.
Current issues of interest to vendors and users of dyes and biological stains. 
| The Biological Stain 
Commission<https://biologicalstaincommission.org/new-dyes/>
Dyes are becoming more expensive! January 2018. BASF, a major dyestuff 
manufacturing company, recently announced that it has increased its prices for 
many pigments and dyes by up to 15% worldwide.
biologicalstaincommission.org
John Kiernan
Secretary. Biological Stain Commission
= = =

From: Charles Riley via Histonet 
Sent: October 9, 2020 10:14 AM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] Alcian Blue staining

We are having an issue where the goblet cells in our control tissue are not 
staining pink even though the patient tissue is staining beautifully on the 
same slide.

The stain is done manually.  What can be some causes for the issues in staining 
on the control section?
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Re: [Histonet] Alcian Blue staining

[Daniel Pesino via Histonet]

Hi Charles - your control tissue may be autolyzed. Try using a well-processed 
control tissue. If that's not it, make sure that your control tissue is being 
completely deparaffinized and that it is being totally covered with the stains.

Good luck!

Daniel Pesino, HTL(ASCP)QIHC CM
Senior Histotechnologist
Trinity Health Of New England
daniel.pes...@trinityhealthofne.org
W  860-714-4675

114 Woodland Street
Hartford, CT 06105
TrinityHealthOfNE.org | Facebook | Twitter | Instagram

-Original Message-
From: Charles Riley via Histonet  
Sent: Friday, October 9, 2020 10:14 AM
To: histonet@lists.utsouthwestern.edu
Subject: [External] [Histonet] Alcian Blue staining

Warning:  This email originated from the Internet!
DO NOT CLICK links if the sender is unknown, and NEVER provide your password.

We are having an issue where the goblet cells in our control tissue are not 
staining pink even though the patient tissue is staining beautifully on the 
same slide.

The stain is done manually.  What can be some causes for the issues in staining 
on the control section?
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[Histonet] Alcian Blue staining

[Charles Riley via Histonet]

We are having an issue where the goblet cells in our control tissue are not 
staining pink even though the patient tissue is staining beautifully on the 
same slide.

The stain is done manually.  What can be some causes for the issues in staining 
on the control section?
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[Histonet] Alcian Blue stain on lab coats

[Eileen Akemi Allison via Histonet]

Hello Histo Peeps:

I have two new lab coats which have been stained with Alcian Blue and can’t get 
the stains out.  I have tried oxy clean incorporated with bleach, as well as 
pre-soaking in 3% acetic acid, and American Mastertech’s stain remover to no 
avail.  Any suggestions would be greatly appreciated.

Thank you in advance,

Akemi Allison, BS, HT/HTL (ASCP)
Pathology Manager
Monterey Bay GI Consultants Laboratory
23 Upper Ragsdale Drive, Suite 200
Monterey, CA 93940
W: Email: aalli...@montereygi.com 
H: Email: akemiat3...@gmail.com 
Tele: (831) 375-3577 X117
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Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

[Hobbs, Carl via Histonet]

Vielen dank, Michael

Very kind of you to send this to me.

Alcian blue is indeed an interesting dye

I thought that it was no longer manufactured?

I am wrong, obviously.

I have used the CEC method many years ago when analysing articular cartilages 
and it is good to see that it is still a valid method to differentiate the 
various acidic mucins


Alles Gute


carl


Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL

020 7848 6813

From: Dr. Michael Gudo (Morphisto GmbH) 
Sent: 24 August 2018 11:22:55
To: Hobbs, Carl
Cc: histonet
Subject: Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

Hello Carl,

Alcianblue is q quite interesting stain! Maybe this selection of a presentation 
helps you to find a suitable method. I attach you a PDF-document to this email. 
The slides are in german, but I think you will understand them anyway.

Kind regards
Michael


Am 23.08.2018 um 20:32 schrieb Hobbs, Carl via Histonet 
mailto:histonet@lists.utsouthwestern.edu>>:

Perhaps Alcian blue at pH3 ( all acidic mucins) on one slide/section  and Ab 
pH1 ( highly sulphated mucins only) on a sequential slide?
Sure, instead  use Alcian blue CEC method?
I will be interested to read other responses to your request.

Mucin-illy

carl




Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL

020 7848 6813
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Weismüllerstr. 45
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Telefon:  069 / 400 3019 - 62
Telefax: 069 / 400 3019 - 64

E-Mail: michael.g...@morphisto.de<mailto:i...@morphisto.de>
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Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

[Hobbs, Carl via Histonet]

You are right!

Thank you for correcting me.

Surely sialomucins will be positive for PAS and H.acid will be negative?


Curious-illy


Carl


Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL

020 7848 6813

From: Mac Donald, Jennifer 
Sent: 23 August 2018 19:52:52
To: Hobbs, Carl
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

The hyaluronidase is digesting out hyaluronic acid, a component found in 
connective tissue mucins.  This reaction is used to differentiate connective 
tissue mucins from epithelial mucins.  Hyaluronic acid and epithelial 
sialomucins will give the same staining results, both positive for alcian blue 
pH 2.5 and negative for pH 1.0.

-Original Message-
From: Hobbs, Carl via Histonet 
Sent: Thursday, August 23, 2018 11:32 AM
To: histonet 
Subject: Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

Perhaps Alcian blue at pH3 ( all acidic mucins) on one slide/section  and Ab 
pH1 ( highly sulphated mucins only) on a sequential slide?
Sure, instead  use Alcian blue CEC method?
I will be interested to read other responses to your request.

Mucin-illy

carl




Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL

020 7848 6813
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Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

[Mac Donald, Jennifer via Histonet]

The hyaluronidase is digesting out hyaluronic acid, a component found in 
connective tissue mucins.  This reaction is used to differentiate connective 
tissue mucins from epithelial mucins.  Hyaluronic acid and epithelial 
sialomucins will give the same staining results, both positive for alcian blue 
pH 2.5 and negative for pH 1.0.

-Original Message-
From: Hobbs, Carl via Histonet  
Sent: Thursday, August 23, 2018 11:32 AM
To: histonet 
Subject: Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

Perhaps Alcian blue at pH3 ( all acidic mucins) on one slide/section  and Ab 
pH1 ( highly sulphated mucins only) on a sequential slide?
Sure, instead  use Alcian blue CEC method?
I will be interested to read other responses to your request.

Mucin-illy

carl


  
 
Carl Hobbs FIBMS 
Histology and Imaging Manager 
Wolfson CARD 
Guys Campus, London Bridge  
Kings College London 
London 
SE1 1UL 
  
020 7848 6813
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Re: [Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

[Hobbs, Carl via Histonet]

Perhaps Alcian blue at pH3 ( all acidic mucins) on one slide/section  and Ab 
pH1 ( highly sulphated mucins only) on a sequential slide?
Sure, instead  use Alcian blue CEC method?
I will be interested to read other responses to your request.

Mucin-illy

carl


  
 
Carl Hobbs FIBMS 
Histology and Imaging Manager 
Wolfson CARD 
Guys Campus, London Bridge  
Kings College London 
London 
SE1 1UL 
  
020 7848 6813
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[Histonet] Alcian Blue digested w Hyaluronidase - any alternatives?

[Cheryl via Histonet]

Help?   we have a student with one of the online schools and we're doing 
specials like it is 1972.  She has to do a Al Blue 2.5 w Hyaluronidase 
digestion.   We haven't done this since the development of clinical IHC two 
dozen years ago -
Is there any other way to acheive this outcome on a colon slide or do I have to 
order the reagent?
Even your best guess is welcome as we didn't get ahead enough to order the 
reagent and she'll end up submitting a null response (and it will be largely my 
fault :(  ) Cheryl Kerry, HT(ASCP) 
ADG Houston Path

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[Histonet] Alcian blue (was Suggestions for staining ground substances ...)

[John Kiernan]

Dear Histonetters,

I recently replied to a question about staining extracellular ground 
substance with alcian blue. Here is some more information.

With the alcian blue-PAS sequence it is necessary to have a good alcian blue 
dye - the same as or functionally equivalent to the original alcian blue 8G. 
For the PAS-alcian blue sequence, other alcian blues may also be OK. 

Alcian blue from a batch certified (or rechecked) by the Biological Stain 
Commission (BSC) 5 or fewer years ago should be good for either procedure. 

Of the dyes commonly used to make up staining solutions for microscopy, alcian 
blue is the only only one that sometimes deteriorates, unpredictably and often 
quite quickly, as a dry powder in the bottle. Some alcian blue powders from the 
1950s are still OK, but some very much younger ones are no good 5+ years after 
passing the BSC's analytical and staining tests. 

John Kiernan
London, Canada
http://biostain.com
= = =
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[Histonet] Alcian Blue

[Diana McCaig]

What is the shelf life of  prepared  1% Alcian blue in 3% Acetic Acid?

Diana
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Re: [Histonet] Alcian Blue

[Rene J Buesa]

Until the staining starts to weaken
René J.

From: Diana McCaig dmcc...@ckha.on.ca
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu 
Sent: Wednesday, February 27, 2013 2:55 PM
Subject: [Histonet] Alcian Blue

What is the shelf life of  prepared  1% Alcian blue in 3% Acetic Acid?

Diana
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RE: [Histonet] Alcian Blue

[Elizabeth Chlipala]

Why would you use a reagent until it starts to weaken, meaning the staining 
intensity and results are changing over time.  In my opinion that's really poor 
quality control.  Stain intensity, specificity and sensitivity should remain 
constant.

Liz

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, February 27, 2013 11:57 AM
To: Diana McCaig; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Alcian Blue

Until the staining starts to weaken
René J.

From: Diana McCaig dmcc...@ckha.on.ca
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu
Sent: Wednesday, February 27, 2013 2:55 PM
Subject: [Histonet] Alcian Blue

What is the shelf life of  prepared  1% Alcian blue in 3% Acetic Acid?

Diana
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RE: [Histonet] Alcian Blue

[Smith, Allen]

Alcian blue lasts until loss of dye onto the sections weakens staining.  If one 
runs many slides through it, it may not last 6 weeks.  With light use, it may 
last 6 years.
Allen A. Smith
Professor of Anatomy
Barry University School of Podiatric Medicine

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Wednesday, February 27, 2013 3:03 PM
To: Rene J Buesa; Diana McCaig; 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Alcian Blue

Why would you use a reagent until it starts to weaken, meaning the staining 
intensity and results are changing over time.  In my opinion that's really poor 
quality control.  Stain intensity, specificity and sensitivity should remain 
constant.

Liz

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, February 27, 2013 11:57 AM
To: Diana McCaig; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] Alcian Blue

Until the staining starts to weaken
René J.

From: Diana McCaig dmcc...@ckha.on.ca
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu
Sent: Wednesday, February 27, 2013 2:55 PM
Subject: [Histonet] Alcian Blue

What is the shelf life of  prepared  1% Alcian blue in 3% Acetic Acid?

Diana
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RE: [Histonet] Alcian Blue

[joelle weaver]

AB is a basic dye that is water soluble, it reacts with anionic groups. The 
classic manual methods I checked use 30 minutes incubation times. What cells 
is your pathologist wanting to highlight, mucins? The general method uses 
acetic, and the methods for sulfated  AMS, uses 1 N HCL. I guess we need to 
know if you are doing the general method and automated or manual- It might be a 
pH issue as much as the staining times. What is your control tissue?




Joelle Weaver MAOM, HTL (ASCP) QIHC
  From: sa...@hotmail.ca
 To: histonet@lists.utsouthwestern.edu
 Date: Thu, 6 Dec 2012 20:58:10 -0500
 Subject: [Histonet] Alcian Blue
 
 
 Hi Everyone:
 Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
 and some say 10 min?Our control works well at 10 minutes but today I had a 
 Dr. say that he expected a small amount of cells to stain and they didn't in 
 an esophagus bx.So, now I'm wondering if b/c most of the procedures that I've 
 read say 30 min, that's what would be best?
 Thanks
 Sheila  
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Re: [Histonet] Alcian Blue

[Lee Peggy Wenk]

I think the key phrase was that the pathologists expected a small amount of 
cells to stain in the esophagus biopsy.


If your control has a lot of acid mucopolysaccharides (AMP) in it (such as 
using a normal intestine, or lung with bronchus), then 10 minutes in Alcian 
blue would definitely show positivity in the goblet cells or mucin cells, of 
which there are a lot of goblet cells in the intestine and mucin cells in 
the bronchus, and each goblet/mucin cell has a lot of AMP.


If on the other hand, there are only a few cells, and particularly if each 
of the cells has a minimum amount of AMP (common in dysplastic or cancerous 
conditions), then 30 minutes in Alcian Blue would be better to demonstrate 
minimal number of cells or minimal amounts of AMP.


Use 30 minutes staining time for Alcian blue all the time, to help find the 
minimal number of cells and/or minimal amounts of AMP.


Peggy A. Wenk, HTL(ASCP)
Beaumont Hospital
Royal Oak, MI 48073

Opinions expressed do not reflect upon Beaumont Hospital.

-Original Message- 
From: Sheila Adey

Sent: Thursday, December 06, 2012 8:58 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alcian Blue


Hi Everyone:
Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
and some say 10 min?Our control works well at 10 minutes but today I had a 
Dr. say that he expected a small amount of cells to stain and they didn't in 
an esophagus bx.So, now I'm wondering if b/c most of the procedures that 
I've read say 30 min, that's what would be best?

Thanks
Sheila  ___
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RE: [Histonet] Alcian Blue

[Susan.Walzer]

We've always used 30 minutes and it has never failed.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila Adey
Sent: Thursday, December 06, 2012 8:58 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alcian Blue


Hi Everyone:
Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
and some say 10 min?Our control works well at 10 minutes but today I had a Dr. 
say that he expected a small amount of cells to stain and they didn't in an 
esophagus bx.So, now I'm wondering if b/c most of the procedures that I've read 
say 30 min, that's what would be best?
Thanks
Sheila
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RE: [Histonet] Alcian Blue

[Lynette Pavelich]

30 here.

Lynette


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] on behalf of 
susan.wal...@hcahealthcare.com [susan.wal...@hcahealthcare.com]
Sent: Friday, December 07, 2012 3:04 AM
To: sa...@hotmail.ca; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Alcian Blue

We've always used 30 minutes and it has never failed.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila Adey
Sent: Thursday, December 06, 2012 8:58 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alcian Blue


Hi Everyone:
Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
and some say 10 min?Our control works well at 10 minutes but today I had a Dr. 
say that he expected a small amount of cells to stain and they didn't in an 
esophagus bx.So, now I'm wondering if b/c most of the procedures that I've read 
say 30 min, that's what would be best?
Thanks
Sheila
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Re: [Histonet] Alcian Blue

[Rene J Buesa]

If after many successful runs with your Alcian Blue reagents you get a 
complaint about either strength of staining or cells not staining at all, most 
likely you have a problem with the solutions. If you prepare them in house 
check the preparation date and prepare a fresh solution. The general procedure 
calls for 30 min, unless you use a microwave oven version of the staining 
protocol.
René J.

From: Sheila Adey sa...@hotmail.ca
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu 
Sent: Thursday, December 6, 2012 8:58 PM
Subject: [Histonet] Alcian Blue


Hi Everyone:
Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
and some say 10 min?Our control works well at 10 minutes but today I had a Dr. 
say that he expected a small amount of cells to stain and they didn't in an 
esophagus bx.So, now I'm wondering if b/c most of the procedures that I've read 
say 30 min, that's what would be best?
Thanks
Sheila                         ___
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[Histonet] Alcian Blue

[Sheila Adey]

Hi Everyone:
Can anyone tell me why some Alcian blue procedures say 30 min in Alcian Blue 
and some say 10 min?Our control works well at 10 minutes but today I had a Dr. 
say that he expected a small amount of cells to stain and they didn't in an 
esophagus bx.So, now I'm wondering if b/c most of the procedures that I've read 
say 30 min, that's what would be best?
Thanks
Sheila
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RE: [Histonet] Alcian blue on the Ventana/Cryoembedding Apparatus

[anita dudley]

allison,  we do all of our alcian blue on the machine.  we love it!!
anita dudley
providence hosp
mobile, ala
 

 From: allison_sc...@hchd.tmc.edu
 To: histonet@lists.utsouthwestern.edu
 Date: Fri, 21 Oct 2011 14:00:44 +
 Subject: [Histonet] Alcian blue on the Ventana/Cryoembedding Apparatus
 
 Hello to all in histo land. For those of you that have the ventana special 
 stains machine, can the alcian blue 2.5 be done on the machine? I am not sure 
 if they have kits for this stain. Also is anyone using the cryoembedding 
 apparatus from pathology innovations? Thanks in advance.
 
 Allison Scott HT(ASCP)
 Histology Supervisor
 LBJ Hospital.
 
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[Histonet] Alcian blue on the Ventana/Cryoembedding Apparatus

[Scott, Allison D]

Hello to all in histo land.  For those of you that have the ventana special 
stains machine, can the alcian blue 2.5 be done on the machine?  I am not sure 
if they have kits for this stain.  Also is anyone using the cryoembedding 
apparatus from pathology innovations?  Thanks in advance.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital.

CONFIDENTIALITY NOTICE:
If you have received this e-mail in error, please immediately notify the
sender by return e-mail and delete this e-mail and any attachments from 
your computer system.

To the extent the information in this e-mail and any attachments contain 
protected health information as defined by the Health Insurance Portability 
and Accountability Act of 1996 (HIPAA), PL 104-191; 45 CFR Parts 160 and 
164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or 
privileged.  This e-mail may also be confidential and/or privileged under 
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above.  If you are not the intended recipient, or any authorized 
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AW: [Histonet] Alcian blue on the Ventana/Cryoembedding Apparatus

[Gudrun Lang]

Ventana has a kit for alcian blue.
I wasn't very satisfied with this kit. The staining was faint in comparison
to manual staining. 
Staining temp is at 60 degrees. I'm not sure how the temp influences exactly
the pH of the staining solution, but it may decrease it under the wanted
2,5. So the result is rather a pH1 than pH2,5 alcian blue.
That are just theoretic considerations, never checked. - what do you think?
Gudrun

-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Scott,
Allison D
Gesendet: Freitag, 21. Oktober 2011 16:01
An: 'histonet@lists.utsouthwestern.edu'
Betreff: [Histonet] Alcian blue on the Ventana/Cryoembedding Apparatus

Hello to all in histo land.  For those of you that have the ventana special
stains machine, can the alcian blue 2.5 be done on the machine?  I am not
sure if they have kits for this stain.  Also is anyone using the
cryoembedding apparatus from pathology innovations?  Thanks in advance.

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital.

CONFIDENTIALITY NOTICE:
If you have received this e-mail in error, please immediately notify the
sender by return e-mail and delete this e-mail and any attachments from 
your computer system.

To the extent the information in this e-mail and any attachments contain 
protected health information as defined by the Health Insurance Portability 
and Accountability Act of 1996 (HIPAA), PL 104-191; 45 CFR Parts 160 and 
164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or

privileged.  This e-mail may also be confidential and/or privileged under 
Texas law.  The e-mail is for the use of only the individual or entity named

above.  If you are not the intended recipient, or any authorized 
representative of the intended recipient, you are hereby notified that any 
review, dissemination or copying of this e-mail and its attachments is 
strictly prohibited.

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[Histonet] Alcian blue-safranin

[Figliuolo, Leticia]

Hello everybody,

I was wondering if anyone has a protocol for Alcian Blue-Safranin (for mast 
cells).

I would greatly appreciate any help.

Thank you!

Leticia Figliuolo
e-mail: leticia.figliu...@roche.com


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[Histonet] alcian blue 2.5 and derm specimens

[faith14913]

We are having a problem with our alcian blue pH2.5 stain.  It is taining the 
collagen really dark and the mucin really light.  My boss just handed me this 
assignment today and told me to find out what is wrong with it, however I am 
not very familiar with this stain.  He suggested I remake all solutions fresh 
and try a known positive control, but I just wanted to check and see what 
histoland thought.

Also for derms, the epidermis on all of our squares and incisions are rolling 
up on the tissue during processing.  It happens when we use the VIP and the 
Pathos (microwave) processors.  I however have never done derms before this job 
so am also not very familiar with itany help???  Thanks!



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[Histonet] Alcian blue quistion

[mohamed abd el razik]

hi every body
i have a confusing quistion reagrding Alcian blue pH 1  2.5 stains.
suppose you have 100 goblet cell react positive to pH 1 which mean all have 
sulfated mucin. and 50 cell from other section react positive to pH 2.5 which 
mean acidic type(sulfated or sialated). why these 50 cell only react to pH 2.5 
althogh the other non reacted cells possess acidic sulfated type?? on other 
word i need to know for what type of mucins alcian B 2.5 react with?
 
thanx all
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[Histonet] Alcian blue quistion

[mohamed abd el razik]

hi every body
i have a confusing quistion reagrding Alcian blue pH 1  2.5 stains.
suppose you have 100 goblet cell react positive to pH 1 which mean all have 
sulfated mucin. and 50 cell from other section react positive to pH 2.5 which 
mean acidic type(sulfated or sialated). why these 50 cell only react to pH 2.5 
althogh the other non reacted cells possess acidic sulfated type?? on other 
word i need to know for what type of mucins alcian B 2.5 react with?
 
thanx all
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[Histonet] Alcian Blue-Acid mucosaccharides with and without hyaluronidase

[Kathy M. Gorham]

Does anyone out there do this stain? Alcian Blue-Acid mucosaccharides
with and without hyaluronidase? Where do you get the testicular
hyaluronidase?  Thank you so much. This for a student.
Kathy Gorham H.T.


GRH National Recognition
Outstanding Rural Health Organization of 2009 awarded by NRHA
Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP
Leader in Innovative Excellence 2009 awarded by the OAHHS
Financial Excellence Award 2010 awarded by the national Rural Health Research  
Policy Analysis Center
Healthcare Achievement Award for Quality in Patient Care Delivery and 
Satisfaction 2010 awarded by Amerinet

GRH Mission
We will ensure access to high-quality, cost-effective health services in a 
safe, customer-friendly environment for all those in need of our services.


GRH Confidentiality Notice
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and should be protected against viewing by unauthorized persons. The information
herein may have been disclosed from records whose confidentiality is protected
by Federal and State Law. Federal regulations prohibit further distribution or
copying of this information without permission.  If you received this e-mail
transmission in error, please notify the sender immediately to arrange for 
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Re: [Histonet] Alcian Blue-Acid mucosaccharides with and without hyaluronidase

[Geoff McAuliffe]

On 3/3/2011 1:47 PM, Kathy M. Gorham wrote:

Does anyone out there do this stain? Alcian Blue-Acid mucosaccharides
with and without hyaluronidase? Where do you get the testicular
hyaluronidase?  Thank you so much. This for a student.
Kathy Gorham H.T.


You can get the enzyme from Sigma. Note that there are two different 
hylaluronidases, bovine testicular and Streptomyces. They have different 
activities.


Geoff


GRH National Recognition
Outstanding Rural Health Organization of 2009 awarded by NRHA
Gold Standard Critical Access Hospital 2009 awarded by LarsonAllen LLP
Leader in Innovative Excellence 2009 awarded by the OAHHS
Financial Excellence Award 2010 awarded by the national Rural Health Research  
Policy Analysis Center
Healthcare Achievement Award for Quality in Patient Care Delivery and 
Satisfaction 2010 awarded by Amerinet

GRH Mission
We will ensure access to high-quality, cost-effective health services in a 
safe, customer-friendly environment for all those in need of our services.


GRH Confidentiality Notice
This e-mail and any attached documents are for the intended recipient/s only
and should be protected against viewing by unauthorized persons. The information
herein may have been disclosed from records whose confidentiality is protected
by Federal and State Law. Federal regulations prohibit further distribution or
copying of this information without permission.  If you received this e-mail
transmission in error, please notify the sender immediately to arrange for 
return
or destruction of this information.

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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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[Histonet] Alcian blue alizarin red double stain

[Thomas, Nancy]

Our lab has a request for alcian blue/alizarin red staining on mouse kidney 
teratomas.  This will be done on FFPE slide mounted sections.  In the past, we 
have done skeletal staining using these stains which also involved all of the 
clearing steps using potassium hydroxide.Has anyone done this stain (minus 
all of the clearing steps) on paraffin sections?  If so, could I get a copy of 
your protocol?
Thank you,
Nancy

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RE: [Histonet] Alcian blue alizarin red double stain

[Walters, Katherine S]

I would also be interested in the protocol.

Katherine Walters
Histology Director
Central Microscopy Research Facilities
85 Eckstein Medical Research Building
University of Iowa
Iowa City, Iowa 52242-1101

phone: # 319-335-8142
fax:  # 319-384-4469

katherine-walt...@uiowa.edu
www.uiowa.edu/~cemrf



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Thomas,
Nancy
Sent: Tuesday, April 20, 2010 12:49 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] Alcian blue alizarin red double stain

Our lab has a request for alcian blue/alizarin red staining on mouse
kidney teratomas.  This will be done on FFPE slide mounted sections.  In
the past, we have done skeletal staining using these stains which also
involved all of the clearing steps using potassium hydroxide.Has
anyone done this stain (minus all of the clearing steps) on paraffin
sections?  If so, could I get a copy of your protocol?
Thank you,
Nancy

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[Histonet] Alcian Blue 2.5

[Laurie Elmgren]

Hi Everyone,

Lately, we have been having a problem with our Alcian Blue. It is fine
up to the washing step. Once it hits the water, the blue fades to
unacceptable. We have tried reducing the water rinse after the blue to
just a quick rinse in d-H20, and it is still poor. The vendor replaced
the kit, and it worked fine the first time, then two weeks later, the
same problem arose. Three different techs have tried the stain three
times. Any suggestions?

 

Laurie Elmgren

Histology Supervisor

Sunrise Medical Labs

240 Motor Pkwy

Hauppauge, NY 11788

(631)435-1515x1108

 

 

This message contains privileged and confidential information intended
only for the use of the addressee named above. If you are not the
intended recipient of this message you must not disseminate, copy or
take any action in reliance on it. 

 

 

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Re: [Histonet] Alcian Blue 2.5

[Geoff McAuliffe]

The pH of the stain may be too high, it should be 2.5-2.6 so check it 
with a pH meter.
If the pH is fine you/your vendor may have a bad batch of dye since 
under normal conditions the stain is difficult to wash out.
Make sure the dye is certified by the Biological Stain Commission. If 
there is no certification tag the dye is not certified.


Geoff

Laurie Elmgren wrote:

Hi Everyone,

Lately, we have been having a problem with our Alcian Blue. It is fine
up to the washing step. Once it hits the water, the blue fades to
unacceptable. We have tried reducing the water rinse after the blue to
just a quick rinse in d-H20, and it is still poor. The vendor replaced
the kit, and it worked fine the first time, then two weeks later, the
same problem arose. Three different techs have tried the stain three
times. Any suggestions?

 


Laurie Elmgren

Histology Supervisor

Sunrise Medical Labs

240 Motor Pkwy

Hauppauge, NY 11788

(631)435-1515x1108

 

 


This message contains privileged and confidential information intended
only for the use of the addressee named above. If you are not the
intended recipient of this message you must not disseminate, copy or
take any action in reliance on it. 

 

 


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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] Alcian Blue 2.5

[John Kiernan]

If the colour washes out of the stained sections, the dye is not alcian blue, 
whatever the label on the bottle might say. Ask for a refund.
 
One of the criteria for certifying alcian blue for use as a biological stain is 
that the soluble dye becomes completely insoluble and cannot be extracted. The 
tests for certification are all published - Biotech. Histochem. 77:237-275, 
2002 - (pp. 241-242 for alcian blue). For an additional insolubilization test 
and revised spectrophotometric criteria for alcian blue, see also Biotech. 
Histochem. 83: 201-203, 2008.
 
If you buy a pre-made solution, check with the vendor that it contains alcian 
blue from a batch of dye powder that has been certified by the Biological Stain 
Commission (BSC). Each batch of a certified dye has a label with a number 
consisting of some letters (indicating the name of the stain and the company 
that submitted samples), a hyphen, and then a number. For example, DcAn-14 was 
a batch of alcian blue certified in 2009 for Dudley Chemical Co. Every bottle 
of certified alcian blue powder has a label with a number of that kind. 
 
Anyone making up a solution of non-certified alcian blue should not be 
surprised if it doesn't work properly as a stain. Unsatisfactory dyes called 
alcian blue have been around for as long as this dye has been in use as a 
microscopical stain (60 years). The criteria for identifying a good batch 
were set out by Scott  Mowry in 1970: Alcian blue - a consumer's guide. J. 
Histochem. Cytochem. 18:842. These criteria are essentially the same as those 
currently used for certification by the BSC.
 
John Kiernan
Anatomy, UWO
London, Canada
= = =
- Original Message -
From: Laurie Elmgren lelmg...@sunriselab.com
Date: Wednesday, March 24, 2010 5:34
Subject: [Histonet] Alcian Blue 2.5
To: histonet@lists.utsouthwestern.edu

 Hi Everyone,
 
 Lately, we have been having a problem with our Alcian Blue. It 
 is fine
 up to the washing step. Once it hits the water, the blue fades to
 unacceptable. We have tried reducing the water rinse after the 
 blue to
 just a quick rinse in d-H20, and it is still poor. The vendor replaced
 the kit, and it worked fine the first time, then two weeks 
 later, the
 same problem arose. Three different techs have tried the stain three
 times. Any suggestions?
 
  
 
 Laurie Elmgren
 
 Histology Supervisor
 
 Sunrise Medical Labs
 
 240 Motor Pkwy
 
 Hauppauge, NY 11788
 
 (631)435-1515x1108
 
  
 
  
 
 This message contains privileged and confidential information 
 intendedonly for the use of the addressee named above. If you 
 are not the
 intended recipient of this message you must not disseminate, 
 copy or
 take any action in reliance on it. 
 
  
 
  
 
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Re: [Histonet] alcian blue van gieson stain

[Anthony Reilly]

Hi Margaret
 
You perform the Alcian Blue first but as with Movatt's pentachrome you need to 
treat the sections after AB with alcoholic ammonia for an hour to convert the 
AB to Monastral Blue as the AB is soluable in picric acid and the Monastral 
blue resistant. Let me know if you need a method
 
regards
Tony
 
 
 
 

Tony Reilly
Chief Scientist, Anatomical Pathology
Pathology Queensland-PA Laboratory
_
Clinical and Statewide Services Division| QueenslandHealth
 
Level 1, Building 15,Princess Alexandra Hospital
Ipswich Road,WOOLLOONGABBA  Qld4102
Ph: 07 3176 2412
Mob: 0402 139411
Fax: 07 3176 2930
Email: tony_rei...@health.qld.gov.au
Web:  www.health.qld.gov.au/qhcss/
 
 


 Perry, Margaret margaret.pe...@sdstate.edu 13/03/2010 12:30 am 
We want to try the double stain alcian blue Van gieson.  Do you stain for the 
alcian blue first and then the Van gieson?  We do both stains but I am unsure 
of how to put them together.
Margaret Perry
South Dakota State University

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[Histonet] alcian blue van gieson stain

[Perry, Margaret]

We want to try the double stain alcian blue Van gieson.  Do you stain for the 
alcian blue first and then the Van gieson?  We do both stains but I am unsure 
of how to put them together.
Margaret Perry
South Dakota State University

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Re: [Histonet] alcian blue van gieson stain

[Rena Fail]

Alcian Blue first



- Original Message 
From: Perry, Margaret margaret.pe...@sdstate.edu
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Sent: Fri, March 12, 2010 9:30:40 AM
Subject: [Histonet] alcian blue van gieson stain

We want to try the double stain alcian blue Van gieson.  Do you stain for the 
alcian blue first and then the Van gieson?  We do both stains but I am unsure 
of how to put them together.
Margaret Perry
South Dakota State University

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pH meter calibration. Was Re: [Histonet] Alcian Blue

[John Kiernan]

A pH meter must be calibrated hourly or daily, not monthly! 
 
In a research environment, with frequently changing chemicals that might damage 
the glass electrode, you need to calibrate against standard (usually bought) 
buffers several times within every day. pH meter electrodes are horribly 
fragile and horribly expensive. Replacing them is a big item in every lab's 
budget. 
 
John Kiernan
Anatomy, UWO
London, Canada
= = =
- Original Message -
From: Diana McCaig dmcc...@ckha.on.ca
Date: Sunday, December 6, 2009 13:49
Subject: [Histonet] Alcian Blue
To: histonet@lists.utsouthwestern.edu

 In the past, when we made up 1% Alcian blue in 3% acetic acid (using
 bottle distilled water), the pH was always 2.5.  We did not 
 have to
 modify it to get the proper pH.  Still using the same 
 powder, we are
 getting a pH of 2.1.  I hate to adjust it because it seems 
 to affect the
 staining.  Any suggestions.  The pH meter is 
 calibrated monthly.
  
 Diana 
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[Histonet] Alcian Blue

[Diana McCaig]

In the past, when we made up 1% Alcian blue in 3% acetic acid (using
bottle distilled water), the pH was always 2.5.  We did not have to
modify it to get the proper pH.  Still using the same powder, we are
getting a pH of 2.1.  I hate to adjust it because it seems to affect the
staining.  Any suggestions.  The pH meter is calibrated monthly.
 
Diana 
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