Re: [Histonet] questions about rhodanine-copper stain

From:Rene J Buesa

I gave up rodamine many years ago because of weak staining and problematic. I switched to Timm's sulfur oxide methods, very reliable and fast (although a little "smelling").
Rene J.

Mildred Fail  wrote:
i keep the rhodanine stock a year. Our protocol is very simple
Rena fail
RHODANINE SATURATED SOLUTION (STOCK)
Absolute Alcohol 100.0 ml
5 p-dimethylaminobenzylidene* 0.2 gm

RHODANINE SOLUTION (WORKING)
Rhodanine Saturated Solution 3.0 ml
Distilled Water 47.0 ml
Shake stock before measuring and mixing solutions.

1. Deparaffinize and hydrate to distilled water.
2. Using plastic Coplin Jars; preheat the Rhodanine solution in 1200 watt microwave on half power 30 seconds then incubate for 5 minutes.
3. Wash well in several changes of distilled water.
4. Mayer's Hematoxylin for 1 minute. 
5. Rinse with distilled water.
6. A quick rinse in 0.5% Sodium Borax.
7. Rinse well with distilled water
8. Deparaffinize, clear, and mount
It will fade if not cover slipped right away

udrun Lang" 10/26/05 08:23AM >>>
I am dealing with the rhodanine stain, because we shall establish it in our
histolab. 

I've read three different protocols (AFIP, Churukian, webpath). 



AFIP and Churukian require sodiumacetat solutions to mix with the
stocksolution, 

the others require just the dilution of the stocksolution in Aqua dest.

Why? Do both versions work?



Churukian demands coverslipping with aqueous medium, the others don't.

Is the staining result soluble in ethanol?



The stability of the 0,2% rhodanine/ethanol stocksolution is described
between one day and three months.

What is the right shelf live?



I hope someone can explain it to me. 





Gudrun Lang



General hospital Linz, Austria



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