[Histonet] Acid-cleaning of slides for metals stains

Lee & Peggy Wenk lpwenk <@t> sbcglobal.net
Sun Sep 16 19:13:04 CDT 2012

For most histology demonstration of iron or copper, we are not doing a 
quantitative analysis (exactly how much is in the tissue). We are usually 
just demonstrating yes there is iron or copper, or no there is not. In the 
case of hemosiderosis, hemochromatosis, or Wilson's disease, we might be 
documenting there is a LOT of iron or copper, but we're don't demonstrating 
a specific amount. If this is what you are doing (yes/no), then if your d. 
water (either deionized or distilled)  has practically no iron or copper in 
it, there is no need to acid-clean glassware or the slides, as long as you 
do the following:
1) Use the slides as they are from the box.
2) Wash the coplin jars in hot water with soap and bleach (or other 
commercial cleaner). After rinsing in hot tap water several times (3-4), 
rinse in d. water several times (3-4). To test to see if all the soap and 
bleach (or cleaner) has been rinsed off completely, touch a pH strip to the 
wet inside of the coplin jar. If the pH turns acidic or basic, then there is 
still soap and/or commercial cleaner in it. Keep rinsing in d. water until 
pH meter remains neutral.
3) Test your d. water once in a while for contaminants. A company called 
HACH has a lot of kits for this, relatively cheap. Can test for resistivity 
or conductivity, which will tell you how many ions are in the water. If 
there are very few, then don't bother testing for which ones (unless you 
HAVE to do the quantitative testing of minerals/metals). If there are a lot 
of ions, then you might want to find out which ones, either by sending out 
to an outside testing company, or buying some of Hach's tests for copper or 
iron (since this is what you are concerned about).

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

The opinions expressed are mine, and do not reflect one Beaumont Hospital.

-----Original Message----- 
From: Esther C Peters
Sent: Sunday, September 16, 2012 3:20 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Acid-cleaning of slides for metals stains

We are going to do special stains for iron (Perl's Prussian Blue for ferric 
iron, Mallory's Method from WebPath: Internet Pathology Laboratory) and 
copper (Rhodanine from Carson and Hladik) in mouse livers. We have on hand 
unopened boxes of plus-charged microscope slides. Do we need to acid clean 
these slides before we put sections on them for these stains (understanding 
that the charge will no longer exist, but the cleaning is more important for 
these stains)?

I would also appreciate any insights about the best acid-cleaning procedure 
for all glassware for these stains. I have used nitric acid in the past, 
swirling it around the staining dishes and covering glass racks in a 
staining dish (how long should this be for?), then rinsing with 
double-deionized water (or would you recommend distilled only?). Thank you!

Esther C. Peters, Ph.D.
Assistant Professor
Department of Environmental Science & Policy
Biology Program/Medical Technology Coordinator
George Mason University
4400 University Drive, MSN 5F2
Fairfax, VA 22030-4444
Office: David King Hall 3057
Phone: 703-993-3462
Fax: 703-993-1066
epeters2 <@t> gmu.edu

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