Csu w1 sora spinning disk scan head

Fluorescence confocal microscopy for Figure 2—figure supplement 1A and Figure 6—figure supplements 1, 2 was performed using a Zeiss Axio Imager with a Yokogawa spinning-disc confocal scanner. Fluorescence confocal microscopy for all other figures was performed using a custom-built inverted Zeiss Axio Observer with CSU-W1 Sora spinning disk scan head (Yokogawa), 1×/2.8× relay lens (Yokogawa), fast piezo z-drive (Applied Scientific Instrumentation), and a iXon Life 888 EMCCD camera (Andor). Samples were illuminated with 405/488/561/637 nm solid-state laser (Coherent), using a 405/488/561/640 transmitting dichroic (Semrock) and 624-40/692-40/525-30/445-45 nm bandpass filter (Semrock), respectively. Images from either microscope were taken with using Slidebook v6.0 software (Intelligent Imaging Innovations) using a 40×–1.3 NA/63×–1.4 NA objective (Zeiss) depending on sample.

Fluorescence confocal microscopy was performed using a ×63, 1.4 numerical aperture objective on an inverted ZEISS LSM 880-AiryScan (Fig. 2e,f and Extended Data Figs. 2f, 5a,b) or inverted Zeiss Axio Observer with CSU-W1 Sora spinning disk scan head (Yokogawa), 1X/×2.8 relay lens (Yokogawa), fast piezo z-drive (Applied Scientific Instrumentation), a iXon Life 888 EMCCD camera (Andor) and a 405/488/561/637 nm solid-state laser (Coherent) with a 405/488/561/640 nm transmitting dichroic (Semrock) and 624–40/692–40/525–30/445–45 nm bandpass filter (Semrock; all other figures). The ZEISS ZEN 3.4 (blue edition) imaging software and Airyscan Processing were used for images captured with the AiryScan. The Slidebook v.6.0 software from Intelligent Imaging Innovations was used for images captured with the Zeiss Axio.