1. Prepare 3 tubes; 1 large and 2 small tubes.
2. Add Minimum Essential Medium ( MEM ) into specimen container about 10 ml. Mix thoroughly.
[ Usually used 10ml pipette to mix medium and semen].
3. Pipette the mixture into large tube.
4. Centrifuge for 10 minutes at 2000 rpm.
5. Remove supernatant, leave only palette.
6. Smach, shake until palette not stick at bottom of tube.
7. Pipette about 0.4ml GIVF solution into first small tube.
8. Pipette palette by using pasteur glass tube to the lower layer of GIVF solution.
Do not let it mix with solution [let it be gel-like paste at bottom of solution]. Leave for 30 minutes by
9. Pipette the top layer of solution carefully and move to another small tube (second small tube).
10. Mix the solution in the small tube.
11. Pipette a drop of solution from the small tube into counting chamber and glass slide.
Observed under microscope.