[Histonet] Yasue's Silver Nitrate-Rubeanic Acid Method for CalciumOxalate

Tony Henwood AnthonyH <@t> chw.edu.au
Wed Aug 1 18:17:48 CDT 2007


This is the one we have used:

Yasue's Method for Calcium Oxalate

Calcium oxalate can be demonstrated in several organs in patients suffering from hereditary or acquired oxalosis. Calcium oxalate is also common in the thyroid in cases of nodular goitre. Oxalates have been identified in giant cells in various granulomatous lesions such as skin granuloma, tuberculous or chronic granulomatous salpingitis, and sarcoid granuloma (Katoh et al 1993).  Oxalosis is also associated with aspergillosis

Under high power calcium oxalate appears as colourless crystals that exhibit brilliant birefringence under polarised light. They stain black-brown with Yasue's method (Yasue 1969), where as they do not stain with von Kossa's or alizarin red S (Katoh et al 1993).

Yasue (1969) Acta Histochem Cytochem 2:83-95.
Katoh et al (1993) Am J Surg Pathol 17(7):698-705.

Fixation:      10% buffered formalin.

Microtomy:     5μm paraffin sections.
Solutions
1.	5% Acetic Acid
2.	5% Potassium Hydroxide
3.	5% Silver Nitrate
4.	Rubeanic Acid Solution:

Prepare a saturated solution of rubeanic acid in 100ml of 70% ethanol
Add 2 drops of concentrated ammonium

5.	2% Methyl Green
6.	50% ethanol

Method:

1.	Dewax and Hydrate sections
2.	Treat with 5% acetic acid 30min.
3.	Wash in water
4.	Place in 5% Potassium Hydroxide 30min.
5.	Wash in water
6.	Rinse in distilled water
7.	Place in 5% silver nitrate 20min.
8.	Rinse in distilled water
9.	Place in Rubeanic acid solution for 1min
10.	Rinse in 50% ethanol
11.	Rinse in water.
12.	Counterstain in 2% Methyl Green 2min.
13.	Dehydrate, clear and mount

Results:

Calcium Oxalate	Brown/black
Nuclei			Green.


Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of djemge <@t> aol.com
Sent: Thursday, 2 August 2007 8:02 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Yasue's Silver Nitrate-Rubeanic Acid Method for CalciumOxalate



Does anyone have the protocol for Yasue's Silver Nitrate Rubeanic Acid method. One of the researchers is doing a calcium oxalate study. I have already given him Pizzolato's, Von Kossa, and Alizarin Red for comparisons. I have heard that the Yasue's method is much more specific. Thanks, Donna

Donna Emge, HT(ASCP)
Northwestern University
Feinburg School of Medicine 
303 E Superior St. Lurie 7-220
Chicago, IL 60611
d-emge <@t> northwestern.edu

________________________________________________________________________
AOL now offers free email to everyone.  Find out more about what's free from AOL at AOL.com. _______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

*********************************************************************
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 Children's Hospital at Westmead

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




More information about the Histonet mailing list