Vsc 08688 camera

After fixation, cells were imaged using Nikon Ti2 inverted microscope. LAMP1 positive puncta and nuclear staining were imaged using the green channel (GFP) and blue channel (DAPI), respectively. GFP images were acquired at 25% LED intensity and 200 ms exposure. DAPI images were acquired at 15% LED intensity and 75 ms exposure settings. Images were acquired using CFI PLAN APO LAMBDA 40X CF160 Plan Apochromat Lambda 40X objective lens, N.A. 0.95, W.D. 0.17–0.25 mm, F.O.V. 25 mm, DIC, Correction collar 0.11–0.23 mm, Spring Loaded, and using Andor Zyla VSC-08688 camera.

All reporter cell lines were seeded in 96-well glass-bottom plates with #1.5 cover glass (Cellvis, P96-1.5H-N). A549 cells were directly plated while U2OS cells were plated after treating the 96-well plate with collagen solution (Gibco, A1048301) to increase cell adherence. Live cell imaging was performed using Nikon Ti2 inverted microscope with an okolab stage top incubator to maintain 37°C and 5% CO₂. Cells were plated at approximately 1.7 X 10⁴ cells per well for 12–18 h prior to performing the experiment. A total of 4–5 positions were imaged in each well at the indicated time using the NIS-Elements AR software. A549 reporter cell line and A549 bulk sorted cells for all chemical treatment experiments were imaged at 25% LED intensity and 200 ms exposure for GFP channel and at 30% LED intensity and 350 ms for TRITC channel images. U2OS reporter cells and A549 reporter clonal cell line for starvation experiments were imaged at 35% LED intensity and 200 ms exposure for GFP channel and at 45% LED intensity and 350 ms for TRITC channel images. Images were acquired using CFI PLAN APO LAMBDA 40X CF160 Plan Apochromat Lambda 40X objective lens, N.A. 0.95, W.D. 0.17–0.25 mm, F.O.V. 25 mm, DIC, Correction collar 0.11–0.23 mm, Spring Loaded, and using Andor Zyla VSC-08688 camera.