VGO 511: Veterinary Andrology and Reproductive Techniques (1+1)

Rose bengal staining method

PROCEDURE

(Preparation of Rose Bengal stain & Staining method)

Instead of eosin-nigrosin stain, Rose bengal stain can also be used for sperm morphology examination.

Preparation of Rose Bengal stain

Rose Bengal - 3 gm
Formalin (37-41%) - 1 ml
Distilled water - 100 ml

• The stain is prepared by adding and grinding the contents in pestle and mortar.

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Staining method

• Prepare a thin smear of semen in a clean grease free slide and dry it
• Place slide in a jar containing Rose Bengal stain for 20 minutes
• Wash the slide in running tap water
• Dry it and examine under oil immersion microscope
• Count 200 sperms and express the results

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