You will need the pre-warmed media, dishes, dispensing pipettor with tips, a pipetting device, a marker pen, timer and cryodevices.
Note that all steps are preformed at room temperature.
First, prepare the dishes.
Dispense 100uL – 1ml Equilibration solutions (ES) into the well/dish.
Dispense 100uL – 1ml Vitrification solution (VS) into the second well/dish.
Transfer the embryo(s) to the ES and equilibrate for 5-15 minutes; bear in mind that the time depends on the embryo stage.
Note: Optimal timings must be confirmed in individual laboratory conditions.
During equilibration, prepare the cryodevice taking care to maintain sterility.
This includes labelling with appropriate witnessing.
At the end of the equilibration time, transfer the embryo(s) to the well/dish containing VS for 1 minute.
During equilibration, prepare the cryodevice taking care to maintain sterility.
At the end of the minute, load the embryo(s) on to the cryodevice in minimal volume. Remove any excess medium.
Transfer to the liquid nitrogen and stir gently.
Cap the device, taking care to keep the sample under the liquid nitrogen.
