RE: [Histonet] Arsenic Laced Chicken/Cherax destructor

From:"Tony Henwood"

This was from Histonet some years ago. The URL is:

Stains for Arsenic
During June 1999, a query was posted to Histonet asking for
demonstration methods for Arsenic in tissues. Following is a summary: 

Roy Ellis (1) and Lynette Thibodeau (2) suggest Castel's Method: 

Fix in 10% formalin containing 2.5% copper sulfate for 5 days. 
Wash for 24 hours in running water. 
Process and embed in parffin wax. 
Deparaffinized sections show green granules of Scheele's green (CuHAsO3)
which, though insoluble in water, is dissolved by acids and by ammonium
hydroxide. By substituting copper acetate for the sulfate, the green
granular paris green or cupric acetoarsenite is produced. Its
solubilities are similar (Castel's method, Bull.Histol.Appliq. V13:106,
1936). A light safranin counterstain gives good contrast. Method
courtesy of Lillie, 3rd edition, Histopathologic technic and practical
histochemistry, page 445. 

John Kiernan (3) writes that arsenic compounds react with hydrogen
sulphide to give insoluble As2S3. This is yellow, and unlikely to be
visible, but could probably be amplified with a physical developer
("autometallography" or Timm's sulphide-silver method). However, this
procedure demonstrates pretty well every metal that has an insoluble
sulphide, so it would have no specificity for Arsenic. 

John also notes that there is a Japanese investigator, Y. Sumi, who has
developed histochemical methods for inorganic substances based on
combinations of chromogenic reagents with mixtures of "masking" agents
that block the reactivity of elements other than the one you want to
stain. His best known methods are for Cd and Hg, but he may have done
something for As. 

Philip Oshel (4) suggests that if the arsenic is expected to be in
nontrace amounts, you can detect, and maybe semi-quatify its presence
with energy-dispersive x-ray spectroscopy (EDX or EDS) in a SEM or TEM.
SEM might be better, as you could use "bulk" specimens--the small bits
of tissue you prepare for sectioning. If there's enough arsenic, you
could use paraffin-embedded sections in the SEM, but be aware that you
could melt the paraffin. This can cause enough contamination in the
column to antagonize the person in charge of the 'scope. 

For TEM, you'd have to use thin sections, but this would give better
localization, if that's important. Prepare the specimen according to
routine procedures for your tissues for either SEM or TEM. 

Roy Ellis (2/6/99) 
Lynette Thibodeau (2/6/99) 
John Kiernan (1/6/99) 
Philip Oshel (1/6/99) 


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-----
[] On Behalf Of Bruce
Sent: Monday, 28 May 2007 12:35 PM
Subject: [Histonet] Arsenic Laced Chicken/Cherax destructor

Dear Colleagues :),
I have an academic who has been feeding her Yabbies (fresh water 
Cray's)/Cherax Destructor  - ...(love the name) with chicken laced 
She is wanting /hoping for - a procedure that will show arsenic in 
the tissue/paraffin sections. I have searched Histology Books & also 
did a Google search to no avail.
Any advice/offers of stains/procedures will be greatly appreciated. Many
thanks in advance, Bruce in OZ
HISTOLOGIST PH:61383446282
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