Setup of Micromanipulator for Sperm Selection and Injection for IMSI: Configuring the Microscope for Intracytoplasmic Morphology-Selected Sperm Injection (IMSI)

Intracytoplasmic Morphologically selected Sperm Injection, or IMSI, involves selecting sperm based on high-magnification, high-resolution imaging prior to microinjection into oocytes. The selection step requires detailed examination of sperm morphology at ~6,000 times magnification and typically utilizes a light microscope configured for differential interference contrast (DIC) imaging with high magnification, high numerical aperture DIC objective lenses, coupled with digital magnification to achieve the necessary resolution and magnification. On the other hand, the post-selection microinjection step utilizes low-magnification Hoffman Modulation Contrast imaging. In this chapter, we will describe the different imaging methods used for sperm morphology examination and microinjection and the microscope hardware required to carry out both steps of the IMSI process. Methods for maintaining an optimal sample temperature and various options for micromanipulators are also reviewed.