-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