Proscan 3

Imaging was performed on a Nikon Eclipse Ti confocal microscope, with a Yokogawa CSU-X1 spinning disk, a Prior Proscan III motorized stage, an Agilent MLC 400B laser launch containing 405, 488, 561, and 650 nm lasers, a cooled iXon DU897 EMCCD camera, and fitted with an environmental chamber to ensure cells were kept at a 37° C and 5% CO₂ during imaging. All images were captured with either a 10X or 20X air objective and were collected at intervals of 2–3 min.

Imaging experiments were conducted on a Nikon Eclipse Ti confocal microscope equipped with a Yokogawa CSU-X1 spinning disk, a Prior Proscan III motorized stage, an Agilent MLC 400B laser launch, and a cooled iXon DU897 EMCCD camera. An environmental chamber was used to maintain cells at 37°C and 5% CO₂ during imaging. In microscopy experiments using optogenetic stimuli, an X-cite XLED1 light source linked to a Polygon400 Mightex Systems digital micromirror device was used to stimulate cells with 500-ms pulses of 450 nm blue light every 1 min, which we define as continuous blue light stimulation. All images were collected using a 20× air, 40× air, or 60× oil objective. Time-lapse images were acquired every 1–3 min.