Stain reagent kit

for cytologyfor sperm analysis

This method is modified from Diff-Quick and Wright-Giemsa stain recommended by WHO. It is intended for staining examination of spermatozoon morphology, sperm cytology and prostate cytology. Principle: Proteins with different isoelectric points in spermatozoon and cells carry different charges under the same pH and thus they will selectively bind to different stains. The amido liberated from acidophilic proteins carry positive charges and bind acid stain (Eosin) to become red. The carboxyls liberated from basophilic protein carry negative charges and bind alkaline stain (Methylene blue) to stain blue. Neutrophil proteins, whose positive charges from the liberated amido are equivalent to the negative charges of liberated carboxyls, bind simultaneously both the acid stain and alkaline stain, and appear mauve. However, as the liberated charges are equal, the stain is weak. The buffer can prevent the interference of acid and alkaline substances for a satisfied result. Methods: Centrifuge the liquefied sperm for 8 minutes (500 rpm) and remove the supernatant. Wash precipitate with isotonic saline solution 2~3 times for 5 minutes each (2,000 rpm). For the specimen with many sperms, take the liquefied sperm from the bottom of tube, wash and centrifuge with isotonic saline solution 2~3 times. Add isotonic saline solution to resuspend the centrifuge precipitate after washing. Push into slice according to the pushing blood film method. Dry in the air or with ventilation. Cover the smear with 1~2 drops of Stain A for 15~20 seconds (the stain may dry at this time). Wash away Stain A with M/15 phosphate buffer, then throw off the buffer briefly.