Histological Staining- Whole Mount Carmine Alum


-Remove mammary gland and flatten/stretch out on a microscope slide


-Air dry tissue 5 min


-Fix overnight in Carnoy’s Solution

            75% ETOH

            25% Glacial Acetic Acid


-Stain overnight in carmine alum

            .2% carmine dye (wt/vol)

            .5% aluminum potassium sulfate (wt/vol)


-Destain (2% HCL in 70% ETOH)---about 2 hours or until background is minimal


-Dehydrate in increasing concentrations of ETOH for 30 minutes each (70%, 80%, 95%, 100%)


-Defat in Toluene and leave in toluene to store or adhere coverslip using permount (sigma)




Carnoys (500 mL)

 375 mL 100% ETOH

125 mL glacial acetic acid


Carmine Alum (250 mL)

.5 g carmine dye (sigma)

1.25 aluminum potassium sulfate (sigma)


Destainer (500 mL)

350 mL ETOH (100%)

28.2 mL of 36-38% HCL

top to 500 mL