Csu x confocal scanning head

Cells were imaged on an inverted Nikon Ti-E (Nikon, Melville, NY) with a Yokogawa CSU-X confocal scanning head (Yokogawa Electric, Tokyo, Japan) and laser merge model with 491, 561, and 642nm laser lines (Spectral Applied Research, Ontario, Canada). Images were collected on a Zyla 4.2 sCMOS Camera (Andor, Belfast, UK). Optogenetic recruitment utilized a Mosaic digital micromirror device (Andor) coupled to a 405-nm laser. A 60× 1.49 NA ApoTIRF oil immersion objective (Nikon) or a 60× 1.2 Plan Apo water (Nikon) objective was used to collect images. MetaMorph Automation and Image Analysis Software (Molecular Devices, Sunnvyale, CA) controlled all hardware.

Optogenetic experiments were performed on an inverted Nikon T-E (Nikon, Melville, NY) with a laser merge module with 491, 561, and 642nm laser lines (Spectral Applied Research, Ontario, Canada) with a Yokogawa CSU-X confocal scanning head (Yokogawa Electric, Tokyo, Japan). The Zyla 4.2 sCMOS Camera (Andor, Belfast, UK) collected the images. Optogenetic activation was achieved using a Mosaic digital micromirror device (Andor) attached to a 405nm laser. Images were collected on a 60X 1.2 Plan Apo water (Nikon) objective. MetaMorph Automation and Image Analysis Software (Molecular Devices, Sunnyvale, CA) controlled all hardware. Fix-and-stain and live-cell imaging of CN03 wash-ins were performed on an LSM 980 system with an Airyscan 2 (Zeiss) detector in super resolution-mode with a 63x NA1.4 oil objective (Zeiss). Microscopy software used was the Zen digital imaging suite (Zeiss).