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

STAINING PROCEDURE AND RESULT INTERPRETATION

Procedure (Video demonstration:

Giemsa staing procedure Part 1 & Giemsa staing procedure Part 2

• A drop of diluted semen (1:5 to 1:10 in 2.9 per cent sodium citrate) is kept on a clean, grease free, pre warmed glass slide and is dried in air. The frozen thawed semen needs no dilution.
• The slide is immersed in 5% formaldehyde for 30 minutes for fixing semen smear.
• The slide is washed in running tap water and dried in air.
• Then the slide is immersed in working Giemsa solution for 3 hours at 37⁰C.
• Finally wash the slide in running tap water and dry it.
• Examine the slide under oil immersion of phase contrast microscope and count 200 sperms.
• The acrosome based on the morphology is classified as follows

S.No.	Defect	Picture
1	Normal acrosome
2	Partially lost acrosome
3	Completely lost acrosome
4	Ruffled/wrinkled acrosome
5	Knobbed acrosome
6	Diadem defect
7	Detached galea capitis
8	Apical notch/ridge

• The fresh semen should have 80 per cent and above acrosome integrity and the frozen semen should have minimum of 65 per cent intact acrosome.
• The hereditary defects like knobbed acrosome and diadem defect should not exceed 5 per cent.

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