Figure 2.

Chilled tethering station for preparing flies. (A) Photograph highlighting the major components of the tethering station: micromanipulator (in black), sarcophagus (in red), Peltier-based chiller with heat-sink (silver and black), and a transparent laser-cut fixture with a hand rest. This setup is typically positioned under a dissecting microscope, and a thermistor is used to measure the top surface of the chiller for closed-loop temperature control. (B) A compact, 3-axis micromanipulator fabricated from 3D-printed parts and simple hardware components. Two different arms are shown, one for the tethering station and the other for the experimental rig. Each axis consists of a rotating handle and screw (in yellow), a locking nut (in red) that fixes the location of the screw relative to the outer rail, and nuts within each carriage (in green), that transfer the linear motion. The device is held together and mounted using additional screws (in purple). (C) A rendering of the sorting and mounting plate, containing a series of indentation, each referred to as a sarcophagus, of different dimensions for different animal sizes. Cold-immobilized flies are sorted on the top section of the plate, and single flies are positioned in one of the cavities for gluing to the tether. (D) A photograph of the plate mounted on top of the chiller with a temperature sensor (yellow tape) and a fly glued to the tether (a dispensing needle). (E) The fly picker used to move anesthetized flies. The picker uses suction controlled by the operator's finger to pick up and deposit single flies.