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Intra Cytoplasmic Sperm Injection
Once the oocyte is mature, it is ready to be fertilized. The sperm sample (can be very poor quality) sperm, frozen, cooled or fresh needs to be prepared. This involves purifying it through a density gradient solution then washing it twice. A plate is prepared with a drop of media for the egg to be placed and a drop of PVP (a very thick liquid for slowing the sperm cell and allowing the injection pipette to damage it's tail then aspirated tail first into the injection pipette. The egg is stabilized with the holding pipette and the sperm is injected.
This procedure allows us to fertilize an oocyte with only one single sperm cell. This is accomplished by maturing the oocyte until it has a polar body, purifying a sperm sample, injecting the sperm into the oocyte, culturing the injected egg until it divides (cleavage). Once it becomes a blastocyst, it is transferred into a recipient mare or vitrified (frozen).
