I need to stain mouse mammary epithelial cells with carmine to detect prolactin. I have the carmine in powder form.
Look into Pearse AGE (1968/1972)Histochemistry, theoretical and applied Vol I and II, presumably the more recent ones are available at your library, also Romeis B (1968) Mikroskopische Technik Oldenbourg Verlag, Munchen, Wien. Romeis is old but you will find recepies for all kind of solutions, how to make and store. Succes.
Preparation
Solution A Carmine stock soluation
Add 2gm carmine, 1gm potassium carbonate, 5gm potassium chloride to 60ml distilled water. Then boil gently for 5 minutes using a large flask to avoid spillage (the heat produces a carmine metallic ion complex which is active staining ingredient. Cool then add 20 cm concentrated ammonia.
Filter and store in dark container at 4 degree, where it should remain usable for several months.
Solution B Carmine working solution.
Stock solution 15ml
Concentrated ammonia 12.5ml
Methanol 12.5ml
Solution C Best's differentiator
Methanol 40ml
Ethanol 80ml
Distilled water 100ml
Method
1- Dewax and positive control sections and brings to water (duplicate sections may be treated with diastase).
2- Stain the nuclei well (with iron haematoxylin stain) differentiate in acid alcohol so that the background is clear- wash and blue.
3- Stain with carmine solution 5-15 minutes (the older stock carmine used the longer staining time).
4- Wash well in either Best's differentiator (solution C) or 74.o.p industrcial methylated sprit.
5- Rinse in fresh alcohol
6- Clear in Xylene and mount as desired.