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