1. Remove any residual cumulus cells from the oocytes by culturing in the hyaluronidase treatment medium (see Subheading 3.2., step 6) for up to 10 min in the hot box until the cumulus cells begin to loosen.
2. Pipet repeatedly with an automatic pipettor to remove the cumulus cells (see Note 11).
3. Wash several times in wash medium to leave the cumulus cells behind.
4. Examine the oocytes under a dissecting microscope while turning the oocytes over to select mature oocytes with polar bodies.
5. Lift these oocytes out of the dish and store in the enucleation medium dish until required.
6. Take a manageable group of oocytes and place in the Hoechst/CB dish and culture for 15 min in the hot box.
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