[Histonet] Pentachrome stain questions

Fri May 23 10:46:50 CDT 2025

Thank you, John. You finally answered my initial question.

In your opinion, does the pentachrome actually give you the old vs new collagen answer? I have a pathologist who orders the pentachrome like most doctors order an H/E stain. I will be doing a lot of these unless I can show a good alternative.

As Always,

Colleen Forster
________________________________
From: John Kiernan <jkiernan at uwo.ca>
Sent: Friday, May 23, 2025 12:02 AM
To: Gudrun Lang <gu.lang at gmx.at>; 'Colleen Forster' <cforster at umn.edu>; Bernice Frederick <b-frederick at northwestern.edu>; Colleen Forster <colleen.forster at biosiminnovations.com>
Cc: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pentachrome stain questions

Why does anyone want to do the "pentachrome" method, which provided 4 (not 5) colours even in a perfectly stained section?  See Lee Luna's "Manual of Histologic Staining Methods of the AFIP" (1968, p.95-97).  Two of the dyes prescribed by HZ Movat (Arch. Path. 80:289-295) in the original 17-step method probably are no longer available.

Russell's modification (21 steps!) is associated with a photo in the late Freida Carson's book. The intense red of a vessel wall makes the tiny surrounding (black) elastin fibres almost invisible. Freida's book provides various better and well illustrated elastin staining methods.

Any correctly used alcian blue dye from a batch recently certified by the Biological Stain Commission will remain in place after minimal treatment with anything slightly alkaline, such as hard tap water.  Check this out at   https://biologicalstaincommission.org/current-issues/
Current Issues | The Biological Stain Commission<https://biologicalstaincommission.org/current-issues/>
Several issues have arisen since 2002, the year of publication of the 10th edition of Conn’s Biological Stains and of a detailed account of tests then used in the Commission’s assay laboratory (Penney et al. 2002), The following notes (alphabetically by names of dyes) refer to new tests, revised standards, and substances added to the list of stains for which certification is available.
biologicalstaincommission.org
John Kiernan
= = =
________________________________
From: Colleen Forster via Histonet <histonet at lists.utsouthwestern.edu>
Sent: May 22, 2025 4:43 PM
To: Gudrun Lang <gu.lang at gmx.at>; 'Colleen Forster' <cforster at umn.edu>; Bernice Frederick <b-frederick at northwestern.edu>
Cc: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pentachrome stain questions

Bernice,

I am aware of this. That is the question.

If in fact, you need that solution to transform the Alcian Blue into a permanent state, how can it work with protocols that don't require that step?

I used the Carson protocol because I was trying several to find one that kept the blue. But I want to understand the "why" here. When you are doing these 100 slides at a time you need it to be reliable and I want to understand.

Colleen Forster
________________________________
From: Bernice Frederick via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Thursday, May 22, 2025 2:22 PM
To: Gudrun Lang <gu.lang at gmx.at>; 'Colleen Forster' <cforster at umn.edu>
Cc: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pentachrome stain questions

[Caution: This email originated from outside the organization.]

I am using the Cancer Diagnostics kit with no issues and it does not use alkaline alcohol at all. Even other procedures I've see do not either. There is one in one of the Freida Carson books that should tell you everything you need to know.
Bernice

-----Original Message-----
From: Gudrun Lang via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Thursday, May 22, 2025 12:59 PM
To: 'Colleen Forster' <cforster at umn.edu>
Cc: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Pentachrome stain questions

Hi Colleen,
can you give us more details of the pentrachrome stain you use? Maybe there is a better chance for help.

Gudrun

-----Ursprüngliche Nachricht-----
Von: Colleen Forster via Histonet [mailto:histonet at lists.utsouthwestern.edu]

Gesendet: Mittwoch, 21. Mai 2025 18:51
An: histonet-request
Betreff: [Histonet] Pentachrome stain questions

Ok special stains guru's, I have a couple questions about reagents for the Pentachrome stain.

Some protocols call for heated alkaline alcohol, some call for this solution at room temp. and some protocols don't use this solution at all.

Can some please help me understand these 3 questions:

   1. What is the purpose of this solution?
   2. Is heat only to accelerate the process or necessary?
   3. How can other protocols not use it at all?

I have been reading papers, surfing the net trying to get a straight understanding. Any help to shed light would be greatly appreciated~
--
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930
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________________________________
From: Colleen Forster via Histonet <histonet at lists.utsouthwestern.edu>
Sent: May 22, 2025 4:43 PM
To: Gudrun Lang <gu.lang at gmx.at>; 'Colleen Forster' <cforster at umn.edu>; Bernice Frederick <b-frederick at northwestern.edu>
Cc: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pentachrome stain questions

Bernice,

I am aware of this. That is the question.

If in fact, you need that solution to transform the Alcian Blue into a permanent state, how can it work with protocols that don't require that step?

I used the Carson protocol because I was trying several to find one that kept the blue. But I want to understand the "why" here. When you are doing these 100 slides at a time you need it to be reliable and I want to understand.

Colleen Forster
________________________________
From: Bernice Frederick via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Thursday, May 22, 2025 2:22 PM
To: Gudrun Lang <gu.lang at gmx.at>; 'Colleen Forster' <cforster at umn.edu>
Cc: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Pentachrome stain questions

[Caution: This email originated from outside the organization.]

I am using the Cancer Diagnostics kit with no issues and it does not use alkaline alcohol at all. Even other procedures I've see do not either. There is one in one of the Freida Carson books that should tell you everything you need to know.
Bernice

-----Original Message-----
From: Gudrun Lang via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Thursday, May 22, 2025 12:59 PM
To: 'Colleen Forster' <cforster at umn.edu>
Cc: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Pentachrome stain questions

Hi Colleen,
can you give us more details of the pentrachrome stain you use? Maybe there is a better chance for help.

Gudrun

-----Ursprüngliche Nachricht-----
Von: Colleen Forster via Histonet [mailto:histonet at lists.utsouthwestern.edu]

Gesendet: Mittwoch, 21. Mai 2025 18:51
An: histonet-request
Betreff: [Histonet] Pentachrome stain questions

Ok special stains guru's, I have a couple questions about reagents for the Pentachrome stain.

Some protocols call for heated alkaline alcohol, some call for this solution at room temp. and some protocols don't use this solution at all.

Can some please help me understand these 3 questions:

   1. What is the purpose of this solution?
   2. Is heat only to accelerate the process or necessary?
   3. How can other protocols not use it at all?

I have been reading papers, surfing the net trying to get a straight understanding. Any help to shed light would be greatly appreciated~
--
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
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_______________________________________________
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