[Histonet] Manual Copper stain

[Matthew Lunetta]

A. EQUIPMENT
· Coplin Jars
· Graduated Cylinders

B. REAGENTS 
RHODANINE SATURATED SOLUTION (stock) 
p-dimethylaminobenxalrhodanine .3 gm 
Absolute alcohol 100 ml
Let the solution sit to allow sediment to fall to the bottom.

RHODANINE SATURATED SOLUTION (working)
Rhodanine saturated solution 3 ml
Distilled water 40 ml
Do not shake stock solution before removing the needed quantity from the
top of the solution. Mix the working solution when placing the slides
into the coplin jar. 

DILUTED MAYERS HEMATOXYLIN SOLUTION
Mayer*s hematoxylin & distilled water (50:50)


0.5% SODIUM BORATE (borax) AQUEOUS
Sodium borate 0.5 gm
Distilled water 100 ml

C. TISSUE CONTROL
Tissue positive for Copper

1. Deparaffinize and hydrate to water
2. Incubate in Rhodanine working solution at 67*C for 1 hour. 
3. Wash well in several changes of distilled water.
4. Stain in diluted Mayers hematoxylin for 10 minutes.
5. Rinse with distilled water.
6. Quickly rinse in 0.5% sodium borate.
7. Rinse well with distilled water.
8. Dehydrate, clear and mount with synthetic media


D. RESULTS
Copper * Bright red to red yellow
Nuclei * Light blue

REFERENCE 
Modified for altitude from the Rhodanine Method for Copper: Theory and
Practice of Histology Sheehan, Hrapchak, Second Edition, pg 230.



Hope this helps,
Matt Lunetta BS HT (ASCP)