[Histonet] Metals

Wed Oct 2 01:49:17 CDT 2019

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Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) | Principal Scientist; Adjunct Fellow, School of Medicine, University of Western Sydney; Visiting Lecturer, School of Life Sciences, Faculty of Science, University of Technology Sydney | Histopathology
t: (02) 9845 3306 | e: tony.henwood at health.nsw.gov.au<mailto:tony.henwood at health.nsw.gov.au> | w: www.schn.health.nsw.gov.au<http://www.schn.health.nsw.gov.au>
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From: John Kiernan [mailto:jkiernan at uwo.ca]
Sent: Wednesday, 2 October 2019 3:01 PM
To: Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP) <jqb7 at cdc.gov>; Tony Henwood (SCHN) <tony.henwood at health.nsw.gov.au>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: Metals

Timm's sulphide-silver method is very sensitive, and modifications (mostly by Danscher) even more so. Sulphide-silver methods detect only those metals that have insoluble sulphides (copper and zinc but not aluminium, in Jeanine's list). It is necessary to fix in a special solution containing hydrogen sulphide - stink and also serious safety precautions!

Tony's mention of Mallory & Parker's fresh hematoxylin stain prompted me to look it up. The 1939 paper is a free PDF download (Google Scholar: Mallory Parker Hematoxylin Stain Metals). Mallory FB, Parker F (1939) Fixing and staining methods for lead and copper in tissues. Am. J. Pathol. 15: 517-522 and Plates 83-85.
The authors noted the importance of fixation (neutral formalin was OK for copper but no good for lead, which needed 95% or 100% alcohol). Like F.B. Mallory's other papers about staining methods, it's rather vague on technical details and has no references.

The late Ralph D. Lillie reported more thorough investigations of staining for metals with haematoxylin in his classic book Histopathologic Technic and Practical Histochemistry (4th and last edn 1976, ISBN 0070378622), giving colours of the complexes with 30 metals introduced into tissues. I have tried Lillie's method on paraffin sections of rat tissues containing a few of these, and it works.  ISBN 9781907904325 (p.333-334)  may be more accessible than Lillie's book, which has become an expensive classic.

For the more specific stains mentioned in Tony's message you need to do some critical reading. The best place to start may be Frieda Carson's Histotechnology textbook. ISBN  978-0891896401.

Enough about metals for now!

John Kiernan
London, Canada
= = =
________________________________
From: Tony Henwood (SCHN) via Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>>
Sent: 30 September 2019 07:22
To: Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP) <jqb7 at cdc.gov<mailto:jqb7 at cdc.gov>>
Cc: Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>>
Subject: Re: [Histonet] Metals

Two good screening stains are Mallory and Parker’s Fresh Hematoxylin Stain for Metals and  Timm’s Silver Sulphide Method for Metals. Malloy's results:
Aluminium       Blue-black
Copper  Greenish-blue
Iron    Blue-black
Lead    Blue
Zinc    Blue

For more specific staining:
Aluminon Stain for Aluminium Hydroxide
Walton’s Stain for Aluminium  (Phloxine binds the aluminium)
Bedrick et al (1986) method for Zinc
Rubeanic Acid Technique for Copper
Rhodanine Technique for Copper

These methods are quite sensitive but  there are some specificity issues. I can provide further details and references if required. Here are some:

Ohtsuki, Y., Yamaguchi, T., Sonobe, H., Takahashi, K., Hayashi, K., Takenaka, A., ... & Terao, N. (1989). Stain Technology: A Simplified Aluminum Stain in Paraffin Sections of Bone from Hemodialysis Patients. Stain technology, 64(2), 55-59.

Walton, J. R., Diamond, T. H., Kumar, S., & Murrell, G. A. C. (2007). A sensitive stain for aluminum in undecalcified cancellous bone. Journal of inorganic biochemistry, 101(9), 1285-1290.

Bedrick, A. E., Ramaswamy, G., & Tchertkoff, V. (1986). Histochemical determination of copper, zinc, and iron in some benign and malignant tissues. American journal of clinical pathology, 86(5), 637-640.

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

________________________________________
From: Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP) via Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>>
Sent: Monday, 30 September 2019 20:49
To: 'histonet at lists.utsouthwestern.edu'
Subject: [Histonet] Metals

Morning all!

I need some advice re: protocols to demonstrate metals in FFPE tissues. Metals such as copper, aluminum and zinc.

Thanks much!

Jeanine Sanders, BS, HT(ASCP), QIHC(ASCP)
Centers for Diseases Control and Prevention
1600 Clifton Road NE
MS H18-SB
Bldg. 18, Rm SB-114
Atlanta, GA 30329
404-639-3590

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