Semen analysis test - Sri Lanka male female infertility treatment ...

to the zona.HUMAN SPERM-ZP BINDING RATIO TEST• The number of sperm bound to the ZP is strongly related to the fertilizationrate, human sperm-ZP interaction tests have been developed usingoocytes that failed to fertilize in vitro.• These oocytes can be used either fresh or after storage in concentratedsalt solutions.• Because the ZP binding capacity is variable, control (fertile donor) and testsperm are labeled with different fluorochromes (fluorescein andrhodamine).• After incubation with equal numbers of control and test sperm, the oocytesare aspirated through a wide bore pipet to dislodge loosely adherentsperm and the numbers of sperm tightly bound to the ZP are counted witha fluorescence microscope.• Results are expressed as a ratio of binding to the ZP of test and controlsperm for four oocytes.• An alternative method is to cut the zonae in half and expose each to testand control sperm (Hemizona assay).HUMAN SPERM-ZP PENETRATION TEST.• It is difficult to determine the number of sperm penetrating into the ZPwhen many sperm are bound to the surface.• The sperm bound to the surface of the ZP can be sheared off byrepeatedly aspirating the oocyte with a pipet with internal diameter lessthan the diameter of the oocyte (120? m).