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Handling of Gametes for In Vitro Insemination in Birds.

Methods in molecular biology (Clifton, N.J.) (2017-08-16)
Shusei Mizushima, Mei Matsuzaki, Tomohiro Sasanami
ABSTRACT

A characteristic biological property of avian gamete (e.g., extremely large egg and polyspermic fertilization) does not allow the direct observation of sperm-egg interactions in vitro, but recent research advances make it possible to manipulate the gamete in vitro. Here, we describe the techniques for the handling of gametes required for in vitro fertilization assay. In addition, we also introduce the procedures for sperm-perivitelline membrane assay, intracytoplasmic sperm injection, and ex ovo culture.

MATERIALS
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Sigma-Aldrich
Hanks′ Balanced Salt solution, 10 ×, Modified, without calcium, magnesium or sodium bicarbonate