IMAGING NEURONAL SUBSETS AND OTHER CELL-TYPES IN WHOLE-MOUNT DROSOPHILA EMBRYOS AND LARVAE USING ANTIBODY PROBES

This chapter discusses the immunocytochemical procedures used to observe specific tissues and cells within whole-mount preparations of Drosophila embryos and young larvae. A special emphasis is placed on providing examples of antibodies and techniques used to examine the patterns of neurons and axons within the developing nervous system. These methods have two primary applications in the study of Drosophila development. First, once a gene of interest has been cloned and antibodies have been raised to the protein product, these immunocytochemical techniques can be used to determine the pattern of protein distribution during embryogenesis and larval development. Second, antibody probes that recognize defined tissues and cells can be used as markers to analyze phenotypes in mutant embryos. The protocol discussed in the chapter, the secondary antibodies are produced in goats and normal goat serum is used as a blocking agent. This ensures that the secondary antibodies will not recognize any of the immunoglobulins in the normal serum being used as a blocking agent. The procedures outlined in this chapter also work well for immunostaining a wide variety of organisms other than Drosophila. © 1994, Elsevier Science Publishers, B.V.