Oocytes Aspiration Plus




IVF for Large Scale Embryo Production PROTOCOLS
















Oocytes Aspiration Plus (OAP)



Oocyte Aspiration Procedure

      1. Aspiration is performed using OAP (oocyte aspiration plus) media. The tube is labeled with cow # and # of oocytes aspirated. The tube is allowed to sit for 10 min following aspiration in a 39 C warming bath. The tube lid is parafilmed before placement in water bath rack.

      2. A filter is placed in a 50 mL conical tube. A transfer pipet is used to remove fluid and oocytes from the bottom of conical tube, this is repeated for 2-3 more times. The filter is rinsed with OAP media to remove blood followed by OWP media. The filter is placed over a searching dish and rinsed into the searching dish with 15-20 mL of OWP media.

      3. The plate is searched and oocytes are transferred into a 35 mm plate labeled with the cow #. The 35 mm contains 3 mL of OWP and is placed on the warming plate next to the scope just prior to searching the grid dish.

      4. Once all the oocytes are transferred to the 35 mm plate, and washed three times in OWP, they are then transferred using a Drummond tool into the 1mL vial of SAR IVM re-parafilmed and quickly placed back into the transport incubator. Vials are placed all the way to the bottom of the incubator using a hemostat. Care must be taken to ensure vials are not dropped into the bottom upsetting the SAR IVM and oil overlay.

      5. Once oocytes are transported back to the lab, loosen caps on SAR IVM vials and place into CO2 5%, 39 C incubator.

      6. Total incubation time of 22-24hrs for bovine IVM.


      If donors were treated with FSH, with many follicles, the follicles may be filled with blood. RLI OAP HD (OAP heavy duty) can be used to reduce the clots during OPU sessions. OAP HD is supplemented a higher concentration of heparin than regular OAP, this can significantly reduce the blood clots and possible loss of oocytes.






    OPU-IVF Protocol (pdf format)