As an added precaution during the preparation procedure, carefully examine each CryoTip outside of the package. Prior to use, all CryoTips should be examined under a suitable magnification (40x power) for possible damage (such as tip breakages or cracks) that may have occurred during transport.
1. Aseptically dispense one (1) 50 μL drop of ES.
2. Transfer embryo(s) (2 maximum), to the ES drop and expose undisturbed for 6-10 minutes.
NOTE: The specimen(s) will shrink and then gradually return to original size, indicating that equilibration is complete.
3. During above equilibration in ES, aseptically dispense one (1) 50 μL drop of VS 2 minutes prior to complete equilibration.
4. Transfer embryo(s) with minimal volume of medium from ES to the VS drop for 30 seconds before loading.
5. Gently pipette embryo(s) once within VS drop to ensure complete rinse with VS.
NOTE: To minimize floating, after 10 seconds pipette the specimen(s) to the bottom center of the VS drop.
6. Load, seal and plunge CryoTip into LN2 within 80 seconds, not to exceed 110 seconds after initial exposure to VS.
7. Refer to CryoTip Loading Protocol diagram and product insert for detailed loading instructions.