Plan apochromat 100 1.4 oil dic

For yeast cells, SD medium containing agar was poured into a Gene Frame (ThermoFisher Scientific) attached to a cover glass. Cells were loaded and the agar pad was sealed with a coverslip. The cells were transferred into an incubation chamber maintained at 30 °C. Images were obtained using an Axio Observer 7 fluorescence microscope (Zeiss), equipped with an Axiocam 506 mono camera and a 20×, as well as 100× oil objective lens (Plan-APOCHROMAT 100×/1.4 Oil DIC). Focus stabilization was done with the Definite Focus module (Zeiss).
For human HEK293 cells, 40,000 cells were seeded into µ-Slide 4 Well chambered coverslips (Ibidi) using medium without phenol red. Cell transfection was performed as described above using medium without phenol red. After 24 h, HEPES pH 7.4 buffer was added to a final concentration of 25 mM and the cells were transferred to a incubation chamber maintained at 37 °C. Images were obtained using an Axio Observer 7 fluorescence microscope (Zeiss), equipped with an Axiocam 506 mono camera, and a 20× objective lens.

Images were mostly obtained using an Axio Observer Z1 fluorescence microscope (Zeiss), equipped with an Axiocam 506 mono camera and a 20×, as well as a 100× oil objective lens (Plan-APOCHROMAT 100×/1.4 Oil DIC) controlled by the Zen blue software. Raw data were collected as Z-stacks and projected using ImageJ (NIH) with manual quantification.

Cells grown on poly-L-lysine-coated confocal dishes were loaded with Fluorochrome-tagged Aβ_1–42 HiLyte Fluor 555 (1 μg/ml, ANASPEC) for 12 h. Cells were then washed twice with PBS and incubated with cell culture medium before light stimulation. Imaging was performed with a Plan-APOCHROMAT 100×/1.4 Oil DIC oil immersion objective on an LSM980 confocal microscope (Zeiss).