I have found the 16 hour copper method works if you drop the slide in
first thing in the morning and finish it about six hours later. Worth a
try.....
denise woodward, Boston, MA


On Wed, 15 Sep 1999 23:15:35 +0000 "Tony Henwood"
<henwood@mail.one.net.au> writes:
> Dear Kathy,
> 
> >      We were just wondering... does anyone have a copper method 
> that does 
> >      not require 18 hours incubation time?  Thanks!
> >
> Try the following:
> 
> GARVEY'S TECHNIQUE FOR COPPER
> 
>           NOTES: 
> 
>           After pretreatment with picric acid to remove calcium and 
> formalin
>           pigment, and exposure to hot sulphuric acid to remove 
> zinc, iron and
>           manganese, the remaining copper is reacted with silver. 
> 
>           SOLUTIONS: 
> 
>           1.   8% Alcoholic picric acid. 
> 
>           2.   10% (v/v) Sulphuric acid
> 
>           3.   0.025% Lithium Carbonate
> 
>           4.   1% Silver Nitrate
> 
>           5.   2.5% Gum Mastic in Absolute Alcohol
>                     Gum Mastic          0.38g
>                     Absolute Alcohol    15ml
> 
>           6.   0.8% Hydroquinone
>                     Hydroquinone        0.2g
>                     Deionised water     25ml
> 
>           7.   Reducing solution
>                     0.8% Hydroquinone   25ml
>                     2.5% Gum Mastic     15ml
>                     1% Siver Nitrate    1ml
> 
>           8.   5% Sodium Thiosulphate
> 
>           9.   Kernechtrott
> 
> 
>           STAINING PROCEDURE:
> 
>           1.   Dewax and hydrate sections.
> 
>           2.   Place in alcoholic picric acid     15min.
> 
>           3.   Wash in water.
> 
>           4.   Place in preheated 10% sulphuric acid  15min  60oC.
> 
>           5.   Neutralise sections in lithium carbonate   2min.
> 
>           6.   Wash in deionised water.
> 
>           7.   Rinse in absolute ethanol. 
> 
>           8.   Treat in gum mastic   5min.
> 
>           9.   Place in developer at 60oC   15min. 
> 
>           10.  Rinse in absolute ethanol. 
> 
>           11.  Wash in water. 
> 
>           12.  Treat with Sodium Thiosulphate     5min
> 
>           13.  Counterstain in Kernechtrott  3min. 
> 
>           14.  Rinse in water, dehydrate, clear and mount. 
> 
> 
>           RESULTS: 
> 
>                Copper and Melanin       black
>                Nuclei                   red
> 
> 
>           REFERENCE: 
> 
>           Garvey, etal.,(1988)  J.Histotechn. 11(4):265-267. 
> 
>  
> Tony Henwood
> Senior Scientist
> Anatomical Pathology
> Royal Prince Alfred Hospital
> Sydney, AUSTRALIA
> 
> http://www2.one.net.au/~henwood
> http://www.pathsearch.com/homepages/TonyHenwood/default.html
> 

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