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Supravital Stains for Determining Viability (Vitality) of Spermatozoa

Description

Supravital stains can help differentiate live from dead sperm. The findings depend on the integrity of the plasma membrane of viable sperm; an intact plasma membrane is able to exclude certain stains. The percent viable sperm correlate with sperm motility.


Indications for performing a supravital stain: when motility is < 40%

 

Stains:

(1) eosin Y (1% eosin Y in isotonic phosphate buffer)

(2) eosin Y-nigrosin (1% eosin Y and 10% nigrosin in distilled water)

 

A small amount of semen is mixed with the stain. A smear is then prepared and air-dryed. The smear is examined with an oil-immersion lens.

 

Stain

Viable Sperm

Nonviable/Dead Sperm

eosin Y

colorless or white (unstained)

pink or red

eosin Y-nigrosin

colorless or white (unstained)

pink or red

 

percent viable sperm =

= (number of viable sperm) / ((number of viable sperm) + (number of nonviable sperm)) * 100%

 

Limitations:

• Supravital stains do not correlate with motility after cryopreservation (Ochninger et al).


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Specialty

ICD-10