Extended Data Fig. 2: Preparation for calcium imaging from the VNC in singing flies.
From: Activity of nested neural circuits drives different courtship songs in Drosophila
a, Equipment. b, A cold-anesthetized fly was placed on a Peltier plate held at 4 °C. Middle and hind legs were cut with forceps around the femur-trochanter junctions. The proboscis was glued to the head capsule. c, Using a manual manipulator attached to the plate holder, the recording plate was positioned above the fly so that the part of the thorax between the forelegs and middle legs is visible through the central hole of the plate. The fly was attached to the plate by applying glue to the coxa and trochanter of the middle legs and femur of the forelegs. The fly was held at room temperature afterward. d, For dissection, the top side of the plate was filled with saline. The cuticle and the sutures covering the pro- and meso-thoracic ganglia of the VNC were removed with fine forceps. Care must be taken not to deform the thorax during dissection, which could significantly reduce singing behavior during calcium imaging.
