Rhodanine Method for Copper

EMS26602

Components:

Rhodamine Saturated Solution
Mayer’s Haematoxylin
Sodium Borate 5% Aqueous Solution

Fixation: 

10% Buffered Neutral Formalin

Embedding: 

Cut paraffin sections at 6 to 10µm. (Thicker sections may stain better)

Staining Procedure:

1. Hydrate slides to distilled water.
2. Incubate in Rhodanine working solution at 37°C for 18 hours. *To prepare Rhodanine Working Solution mix: Rhodanine Saturated Solution 6ml Distilled water 94ml. Note: Shake stock solution before measuring and mixing solutions and shake working solution when pouring it on slides.
3. Wash well in several changes of distilled water.
4. Stain in diluted Mayer’s Haematoxylin for 10 minutes. (To prepare diluted Mayer’s Haematoxlin mix - Mayers Haematoxylin 50ml Distilled water 50ml)
5. Rinse with distilled water.
6. Quickly rinse in Sodium Borate, 5%.
7. Rinse well with distilled water.
8. Dehydrate through 95% alcohol to absolute alcohol. Clear in Xylene, 2 changes each, and coverslip using a synthetic mounting medium.

Note: The use of chemically clean glassware is necessary. Shake stock solution before measuring and mixing solutions and shake the working solution before pouring it onto the slides.

Results:

Copper – orange/red

Nuclei – light blue

Note: With low copper concentrations in tissue, slight fading may occur after coverslipping and the golden precipitate may be difficult to distinguish from lipofuscin.

References:

Sheehan and Hrapchack, Theory and Practice of Histotechnology. St. Louis, The Mosby Company 1980 p. 230.