Hello Prakash, 

To the best of my knowledge, i suggest that u go for swim up protocol of sperm preparation  for IVF and u can bypass the need for 2 washing's. This method will surely give you >85% live, healthy sperms for IVF. 

all the best! 

Swim-up is one option, perhaps you can also follow a Density gradient separation technique.

Thanks for suggestions 

we are going for swim up procedure and getting good results in cattle sperm but for buck sperm during the swim up protocol at time of incubation with working BO media we are getting lots of dead sperms or exhausted sperms. could this be due to improper cryopreservation or due to our media any suggestions?

Dear Prakash

there is a new product for the magnetic depletion of apoptotic sperm cells:  MACS ART Annexin V Complete Kit (200-070-507). The protocol is fast and simple and should be done after DGC.

If you send me an email ([email protected]) I'll be happy to send you a visual overview of the protocol as well as references.

thanks  Dr. Navid we completed this .. we used swim up approach and reached to late morulla stage ... what we did is Zona thinning by using Acid Tyode medium and we got some  results...