Drosophila Larva Imaginal Disc Dissection: A Method to Observe Developing Epithelia

- To begin the dissection, use one pair of forceps to grasp the larval body in the middle, and a second pair to tear the larva in half. Then pinch the mouth hook at the anterior end while holding the body at the posterior side. Next, push the forceps holding the mouth inside the body. Use the posterior forceps to fold the body wall inside out.

In this preparation, the salivary glands and the gut can be easily identified and removed to reveal the brain and imaginal discs. The antennal, eye, leg, wing, and haltere discs can be seen. Once fixed, transfer the larval preparation onto a slide with mounting medium where the discs can be further dissected. In the example protocol, we will see how to dissect wing imaginal discs using this larval preparation.

- To begin, use a wooden stick or blunt forceps to transfer wandering third instar larvae into a Petri dish with PBS. Then genotype them under fluorescent light to identify the appropriate fluorescent markers for mosaic larvae. Before proceeding, wash the larvae with PBS. Then under a dissection microscope, use two pairs of forceps to pinch the center of the larva, and tear the body in half. Next, pinch the mouth hooks while pushing the