[Histonet] RE: Churukian Silver stain for Fungus

Mon Jun 9 19:40:27 CDT 2014

Hi Terri,

I agree, Churukian's Ammoniacal silver method is very good.

I assume you are referring to the method given in Churukian, C. J., & Schenk, E. A. (1988). Staining Pneumocystis carinii and fungi in unfixed specimens with ammoniacal silver using a microwave oven. Journal of Histotechnology, 11(1), 19-21.

In their discussion they note:
"Paraffin sections can also be stained with this method with only minor changes. In place of 5% periodic acid use 5% chromic acid"
Though the reason given is that paraffin sections tend to detach from slides.

Charles repeats this recommendation in his later article: Churukian, C. J. (1996). Consistent silver methods for demonstrating basement membranes, reticulum, and fungi. Journal of histotechnology, 19(3), 211-217.

I follow Charles recommendations for paraffin sections (ie chromic acid).

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior 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: Terri Braud [mailto:tbraud <@t> holyredeemer.com] 
Sent: Friday, 6 June 2014 10:50 PM
To: Tony Henwood (SCHN)
Subject: RE: Churukian Silver stain for Fungus

Tony -
I totally agree with you (and your references) that Periodic Acid is not as good when used with Grocott's methenamine silver as in the GMS.
However, it totally rocks when used with the Churukian Ammoniacal Silver Method.  We are still talking about 2 different staining methods.
Terri

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874

-----Original Message-----
From: Tony Henwood (SCHN) [mailto:tony.henwood <@t> health.nsw.gov.au]
Sent: Thursday, June 05, 2014 9:17 PM
To: Terri Braud; histonet <@t> lists.utsouthwestern.edu
Subject: RE: Churukian Silver stain for Fungus

Terri,

I was referring to the incorrect use of periodic acid in the GMS stain.

The Grocott's Methenamine silver stain uses chromic acid, not Periodic acid.
There are many instances in the literature where using a classic GMS has been found superior for demonstrating fungus rather than PAS. 
 I am concerned that Periodic acid - silver staining has not been validated using cases where classic GMS has been shown to be more sensitive than PAS (see below) or in cases of pseudofungi, which are PAS positive but overall GMS negative.

An appreciation of the difficulties in relying on Periodic acid rather than chromic acid for GMS staining can be seen in these articles: 
Chandler, F. W., & Watts, J. C. (1995). Fungal diseases. Journal of Histotechnology, 18(3), 247-252

Carson, F. L., Fredenburgh, J., & Maxwell, J. E. (1999). Inconsistent detection of histoplasma capsulatum with periodic acid oxidation in the Grocott methenamine-silver nitrate (GMS) fungus stain. Journal of histotechnology, 22(2), 119-122.

Carson, F. L., & Richmond, R. S. (2010). Periodic acid cannot replace chromic acid in Grocott's method for fungi. Biotechnic & histochemistry:
official publication of the Biological Stain Commission, 85(4), 270.

Henwood, A. F., Prasad, L., & Bourke, V. M. (2013). The application of heated detergent dewaxing and rehydration to techniques for the demonstration of fungi: a comparison to routine xylene-alcohol dewaxing.
Journal of Histotechnology, 36(2), 45-50.

Song, D. E., Kahn, A. G., Khang, S. K., & Ro, J. Y. (2005). Pseudofungi in pericolic lymph nodes. Archives of pathology & laboratory medicine, 129(4), e97-e100.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior 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: Terri Braud [mailto:tbraud <@t> holyredeemer.com]
Sent: Thursday, 5 June 2014 11:29 PM
To: Tony Henwood (SCHN); histonet <@t> lists.utsouthwestern.edu
Subject: RE: Churukian Silver stain for Fungus

I beg to differ.  I think you are confusing stains. I am not talking about using a Periodic Acid Schiff's stain (PAS) to stain fungus.  I am referring to the Churukian Microwave Ammoniacal Silver stain for fungus.
It oxidizes with Periodic Acid and the Silver solution is similar to what is used for most Reticulum stains. It stains the exact same organisms as a Grocott's Methenamine Silver (GMS), just without staining the elastic fibers. Our pneumocystis stains using the Churukian Ammoniacal Silver stain are just beautiful.
This method has been taught at NSH workshops and has been widely used in published literature.  As soon as I can figure out how to post pictures, I will send some pictures of a a pneumocystis control and an aspergillus control stained with the Churukian method.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874

-----Original Message-----
From: Tony Henwood (SCHN) [mailto:tony.henwood <@t> health.nsw.gov.au]
Sent: Wednesday, June 04, 2014 7:20 PM
To: Terri Braud; histonet <@t> lists.utsouthwestern.edu
Subject: RE: Acid Clean Glassware

No,
Using Periodic acid instead of chromic acid just gives you a PASM. 
Pseudo-fungi are PAS Positive but classic GMS (using chromic acid) negative.
The literature is quite confusing on pseudo-fungi. Some say that they are GMS positive whereas other claim they are GMS negative. 
My own experience is that they are GMS (using chromic acid) negative. 
It is possible that our pathologists aren't aware that the "GMS" that their lab does might be using Periodic acid instead of Chromic acid. 
Commercial kits (eg Sigma and Richard-Allan) use periodic acid instead of chromic acid.
The literature often does not report the exact GMS used which makes clear understanding of the histochemical results difficult.

Pneumocystis will not be easy to see unless chromic acid is used (the mucin stains strongly PAS (and hence PASM) positive obscuring the small microorganisms).

Using PAS, Old fungi, Mucor, Actinomyces and Nocordia do not stain well whereas they stain quite well with GMS.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior 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: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Terri Braud
Sent: Thursday, 5 June 2014 5:58 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Acid Clean Glassware

Chromic acid does a nifty job of removing metal deposits on glassware, but so do many commercial lab detergents.
Chromic acid is the oxidizer for the fungus in the GMS stain.  Go one better and get rid of Chromic Acid out of your lab.  It is probably one of the more toxic / nasty chemicals in your department.  Instead, try Churukian's Ammoniacal Silver for Fungus in the microwave.  It is a much simpler, faster, prettier stain.  It uses Periodic Acid as the oxidizer and does not stain the elastic fibers like a regular GMS.  Both you and your pathologists will love it, I promise.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874
Today's Topics:

   1. Acid Cleaned glassware (Abbott, Tanya)
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Please notify the sender of any error by E-Mail.

Thank you for your cooperation.

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This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Sydney Children's Hospitals Network.

This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Sydney Childrens Hospital's Network accepts no liability for any consequential damage resulting from email containing computer viruses.
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