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.
Polygon400 systems
Manufactured by Mightex
The Polygon400 by Mightex Systems is a programmable multi-channel LED light source designed for precision illumination and optogenetic applications. It features up to 400 independently controllable LED channels, allowing for complex spatiotemporal light patterns. The device is configurable and can be customized to meet the specific requirements of various research and industrial applications.
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2 protocols using polygon400 systems
1
Confocal Microscopy with Optogenetic Stimulation
2
Confocal Microscopy with Optogenetic Stimulation
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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.
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