Methods to study meiosis in insect spermatocytes

This chapter covers methods that are useful for the in vitro culture and live-cell study of insect spermatocytes in general and of crane-fly spermatocytes in particular. The merits of crane-fly spermatocytes are detailed in the Introduction section. In the following sections, step-by-step instructions are given for optimizing visualization of meiotic events taking place within living spermatocytes by employing microaspiration to flatten cells and then in subsequent operations to manipulate them via microinjection. Emphasis is on the attributes of ionophoretic injection as a way of introducing fluorescently conjugated proteins into the cytoplasm of flattened spermatocytes. In the last section of this chapter, the presentation of pressure injection is an alternative for delivering cell permeable probes into the interstitial space surrounding spermatocytes within in vitro preparations.