Dfc900 gt camera

A Leica DMi8 Live Cell Imaging System, equipped with an OKOLab stage-top live cell incubation system, LASX software, Leica HCX PL APO 63x/1.40–0.60na oil objective, Lumencor LED light engine, CTRadvanced+ power supply, and a Leica DFC900 GT camera, was used to acquire images. Exposure times were set at 50 ms (GFP, 470 nm) and 200 ms (mCherry, 550 nm), with LED light sources at 50% power, and images acquired every 30 seconds over a 10 min time course. TIRF imaging was conducted using a 100X TIRF compatible objective and 10% laser power.

HEK293T cells and Neuro2a cells were imaged on a Leica DMi8 Live Cell Imaging System, equipped with an OKOLab stage-top live cell incubation system, LASX software, Leica HCX PL APO 63×/1.40 to 0.60 numerical objective oil objective, Lumencor LED light engine, CTRadvanced+ power supply, and a Leica DFC900 GT camera. The mCherry channel (553 nm, 50% power, 200 ms exposure) was used for pre, during, and post-light visualization. GFP channel (480 nm, 50% power, 50 ms exposure) was used for Cry2 activation. Images were captured in 30 sec intervals.
Live-cell confocal images of HEK293T cells were collected on Zeiss LSM 700 laser scanning microscope equipped with a stage-top incubator and ZEN Black 2012 software. Images were captured on the mCherry channel (553 nm) and GFP channel (480 nm) for blue light illumination in 30 sec intervals. For ERK-KTR cell experiments, ERK-KTR was visualized on the GFP channel (5% power), and the nuclear stain was visualized on the DAPI channel (395 nm, 5% power). All images were analyzed using Fiji Image J software.