Centrifuge tube size: 50 ml
Pre-warm centrifuge shields, centrifuge tubes and skim milk extender to body temperature (37° C.)
- Pour 20 ml raw semen in each centrifuge tube.
- Add 20 ml skim milk extender pre-warmed to 37° C (body temperature). Total volume extended semen = 40 ml per tube
- Replace caps, then invert tubes gently 2-3 times.
- Place tubes in centrifuge shields, then place in centrifuge.
CAUTION - make sure tubes within the centrifuge are balanced.
- Set timer for 10-12 minutes.
- Set speed at 1000 rpms or 400 g's. Click here to for RPM calculator
- Allow centrifuge to slow without braking.
- Remove tubes from centrifuge.
- Pour off supernatant from each tube to 5-10 ml. and re-suspend pellet of sperm by gently agitating.
- Add enough extender to bring concentration of progressively motile cells to 25-50 million/ml. Use worksheet to calculate volume of extender. Divide volume by number of centrifuge tubes used.
- Volume of Centrifuged Semen: ml.
- Progressive Motility: %
- Count: million/ml.
|A.||Total Cells (Count x Volume) / 1000 = billion cells.|
|B.||Assume 70% recovery from centrifuge:|
(.70 x Total Cells) = billion Total Cells after centrifugation.
|C.||Divide Total Cells after Centrifugation (B) x 1000 by 25-501. to get Total Extended Volume.|
(B x 1000) / 25-50) = ml. of semen + extender.
|D.|| (B) / 1 billion = Cooled breeding doses: doses.|