Dissection and Immunolabeling of the Central and Peripheral Nervous System of Drosophila Larvae

The ability to visualize the cells and proteins of a tissue within their original context (i.e., in vivo) is invaluable for the study of that biological system. Visualization is especially important in tissues with complex and convoluted structures, such as the neurons and glia of the nervous system. The central and peripheral nervous systems (CNS and PNS, respectively) of the third-instar larvae of the fruit fly, Drosophila melanogaster, are found on the ventral side of the larvae and are overlaid by the rest of the body tissues. Careful removal of overlying tissues while not damaging the delicate structures of the CNS and PNS is essential for proper visualization of these tissues. This protocol describes the dissection of Drosophila third-instar larvae into fillets and their subsequent immunolabeling to visualize endogenously tagged or antibody-labeled proteins and tissues in the fly CNS and PNS.