H.O.S.T (Hypo Osmotic Swelling Test)

TAKE 500 MICRO LITRE HTS-H.O.S. SOLUTIONS IN A SMALL TEST TUBE. ADD 100 MICRO LITER OF LIQUIFIED SEMEN SAMPLE TO THE H.O.S. SOLUTIONS AND MIX WELL. KEEP AT 37 DEGREE CENTIGRADE FOR 30 MINUTES.
ADD 50 MIRCO LITRE OF STOP SOLUTION AND TAKE JUST ONE DROP ON THE PLAIN GLASS SLIDE PUT THE COVER SLIP AND EXAMIN UNDER MICROSCOPE IN 40X OBJECTIVE, CALCULATE PERCENTAGE OF COILED TAIL.
NORMAL PERCENTAGE > 60%.

A.I.S.T (Acrosome Intactness Slide Test)

TAKE 500 MICRO LITRE OF HTS - ACROSOME SOLUTION IN A TEST TUBE, ADD 100 MICRO LETER LIQUIFIED SEMEN SAMPLE TO THE CENTRIFUGE TUBE AND MIX WELL.INCUBATE AT 37DEGREE FOR 30 MINUTES. PUT A DROP OF MIXTURE SOLUTION ON THE HTS-GELATIN COATED SLIDE & MAKE SMEAR VERY SMOOTHLY, AIR DRY FOR 2 MINUTES TO REMOVE EXCESS MOISTURE.
INCUBATE FOR 2 HOURS AT 37 DEGREE CENTIGRATES WITH MOISTURE CHAMBER.
AIR DRY AND EXAMINE UNDER 10X - 40X OBJECTIVE.COUNT THE SPERMATOZOA WITH HALOS.
NORMAL PERCENTAGE > 50%

N.C.D.T (Nuclear Chromatin Decondensation Test)

TAKE 100 MICRO LITRE LIQUIFIED SEMEN IN A TEST TUBE, ADD 1ML. MICROLITER WASH SOLUTION AND MIX WELL AND CENTRIFUGE FOR 5 MINUTES AT 1500 RPM. AFTER CENTRIFUGATION DISCARD ALL THE SUPERNATANT WITHOUT DISTURBING PELLETE.
DISOLVE THE NCD POWDER IN 10 ML DISTILLED WATER PROPERLY.
ADD 200 MICRO LITRE HTS-N.C.D. SOLUTIONS TO THE PELLETE AND MIX WELL. KEEP AT ROOM TEMPERATURE FOR 30 MINUTES.
ADD 50-100 MICRO LITRE OF STOP SOLUTION AND TRANSFER A DROP OF MIXTURE SOLUTION ON THE GLASS SLIDE AND COVERED WITH A COVERSLIP. EXAMINE UNDER 40X OBJECTIVE, COUNT THE NUMBER OF CONDENSED AND DECONDENSED HEAD.
NORMAL PERCENTAGE > 70% DECONDENCE SPERMATOZOA.