Ds fi2 epifluorescence microscope

Images were acquired on an Olympus DS-Fi2 epifluorescence microscope, using 40X and 100X Olympus UplanFI oil immersion objectives, a Nikon DS-fi2 camera and the Q-Imaging capture software. Confocal acquisition was carried out at Unidad de Microscopía Avanzada (UMA), Pontificia Universidad Católica de Chile. Cells were imaged on a Nikon Eclipse C2 Si Confocal Spectral Microscope with NIS-Elements C software. High-resolution confocal images were acquired at UMA and Massachusetts General Hospital (MGH) Program in Membrane Biology (PMB) Microscopy Core Facility, using the Zeiss LSM 800 Airyscan confocal microscope and Zeiss LSM 880 Airyscan confocal microscope, respectively, with Airyscan acquisition mode and conventional super-resolution processing using the ZEN software by Zeiss.

Cells were fixed using 4% paraformaldehyde for 10 min at room temperature. After fixation, three washes with PBS 1× solution were performed, and a final wash in distilled water was made to remove excess salt. Finally, the coverslips were mounted with a Vectashield/DAPI solution (Vector). Images were acquired in dark and light conditions with a DS-Fi2 epifluorescence microscope (Olympus, Shinjuku-ku, Tokyo, Japan) equipped with a Nikon DS-Fi2 camera (Nikon, Minato-ku, Tokyo, Japan) operated with QCapture Suite PLUS 3.1.3.10 (Q-Imaging).