Table 1.

Troubleshooting guidance

Step	Problem	Possible reason	Possible solution
Preparation/running experiments	Worms leaving the wells	Male worm Hyperactive hermaphrodite	Fill moats around the wells with copper sulfate
Constructing the Blue Light Stimulus for Sleep Perturbation, 12	The power supply is turned on but the LEDs do not turn on	Voltage is set too low Wired LEDs may be improperly connected	Increase the voltage Using a multimeter, check the resistance between the positive terminal of the first LED and the negative terminal of the second LED. Properly wired LEDs should have a finite resistance between the input and output terminals.
Setting Imaging System Parameters, 38	The image is too dark	Exposure time is too short Aperture diaphragm is too small LEDs not properly positioned and are too far from sample Voltage of the power supply is too low	Increase the exposure time (Device properties- >Exposure time). Should be less than 1 second, longer exposures result in motion blurring due to animal movement. Open aperture Re-position LEDs Increase the voltage of the power supply
Setting Imaging System Parameters, 38	A white ring is visible at the perimeter of the wells	Scattering from the illumination LEDs	Ensure the illumination LEDs are not pointing upward but instead are directed horizontally, parallel to the sample plane. The angle between the illumination LED direction and the sample should be minimized to reduce scattering of illumination LEDs onto the camera sensor.