PREPARATION AND ANALYSIS OF PURE CELL-POPULATIONS FROM DROSOPHILA

This chapter focuses on the isolation and analysis of pure populations of Drosophila cells. The chapter discusses, whole animal cell sorting (WACS), a relatively new technique that is used to isolate and study a number of different early embryonic cell types in vitro. It can be used to prepare analytical quantities of almost any early embryonic cell type, provided a lacZ strain that expresses β-galactosidase specifically in the cells of interest is available. WACS is used to purify cells from early stages of embryogenesis; applying WACS to later stages of development will probably require the use of other methods for cell dissociation besides the mechanical procedure as most cells become more adherent as the embryo matures. Although traditional methods such as treatment with trypsin or ethylene glycol tetraacetic acid (EGTA) can be used, we can also envision the introduction of a transgene that can be conditionally activated to disrupt cell associations when needed. It is necessary to determine whether endogenous β-galactosidase activity is present at these later developmental stages, which could interfere with the procedure unless it can be specifically inhibited. © 1994 Elsevier Science Publishers, B.V.