Ix83 zdc2

All images were recorded in a modified Olympus IX83 (Tokyo, Japan) with a 100 × 1.4 NA objective and an Andor Zyla 4.2 sCMOS camera with an effective pixel size of 65 nm in the images. CoolLED pE-4000 was used as the light source. Olympus IX83-ZDC2 was used to correct focus drift and an Olympus IX3-SSU ultrasonic stage was used for multipoint imaging. An Olympus double lamp housing U-DULHA was used to introduce a 405-nm diode laser used in microirradiation. An Okolab UNO (Naples, Italy) live-cell chamber supplied with humidified CO₂ and an objective heater was used.

All widefield imaging was performed using an inverted microscope (IX83-ZDC2; Evident/Olympus) equipped with a cMOS camera (Orca-Fusion, Hamamatsu) and Xenon light source. Images were acquired using a 100× 1.59 NA TIRF objective (Olympus) and with Cellsens software (Evident/Olympus).
Confocal microscopy was performed on an inverted laser scanning confocal microscopy (LSM780 or 980, Zeiss). The LSM780 was equipped with a motorized X,Y stage, Z-focus, five lasers (405, 488, 561, 594, 633), and three fluorescence detectors (two flanking PMTs and a central 34 channel GaGasp with ~40% QE). Images were acquired with a 63x Plan-Apochromat oil objective (NA=1.4) using ZEN Black v 2.3. The LSM980 is equipped with a motorized X,Y stage and Z focus with high speed Piezo insert as well as four diode lasers (405, 488, 561, 633). Images were acquired with 1x Nyquist sampling using a 63× 1.4 NA Plan-Apochromat oil objective (Zeiss) and 4 channel GaAsP detectors on Zen Blue 3.6 software. This microscope is equipped with Airyscan 2, however, this module was not used in these experiments.