Figure 6.

Sample experiment settings file

These sample settings define imaging resolution (bin 2), framerate (100 ms exposure = 10 frames/s), and illumination timing (20 ms for brightfield, 10 ms for fluorescence with 5 ms delay from exposure onset). Stimulation recordings capture 300 frames (or 30 s at 10 fps), and the stimulation program is a 10 s chemical pulse (valve 1 opens from frame 50–150 or from 5–15 s within each recording trial). The entire experiment lasts 12 h. Sleep state changes are monitored using a 10-s interval and changes must persist for 3 intervals (i.e., 30 s) to register a transition. After each stimulation trial, 15 min should elapse before triggering a new stimulation (regardless of any transitions within this time delay), and the next trigger shall begin following the opposite transition (i.e., Sleep-to-wake follows Wake-to-sleep).