Vitrification with microinjection of single seminiferous tubules

Abstract This is a protocol for vitrification with microinjection of single seminiferous tubules an efficient cryopreservation approach for limited testicular tissue. Microinjection of seminiferous tubules maintained tubule structural integrity and germ cell numbers, and reduced spermatogonial apoptosis after cryopreservation. But it cannot be used on large testicular tissues. The key stages of the protocol is vitrification solution introduced into the seminiferous tubules via microinjection needles connected to a microinjection pump with a flow rate set at 2 μl/s under microscopy. The time taken for it to be completed is about half an hour for vitrification and 15 min for thawing