Airy fast detector module

Unexpanded and expanded specimens in 35 mm glass bottom dishes were fixed as described in the previous protocol. Wide-field fluorescent images were collected onto an EMCCD camera (Evolve 512, Delta Photometrics) with an oil-immersed objective (150 × /1.45, Olympus), yielding a pixel size of 106 nm. Emissions were collected through an objective and filtered by a 520 nm band-pass filter (FF01-520/35-25, Semrock) for Alexa488 or by a 655 nm band-pass filter (FF02-655/40-25, Semrock) for Alexa647. Airyscan imaging was performed using a commercial microscope (ZEISS, LSM880, Germany) with an additional Airy FAST detector module (Zeiss), equipped with a Plan-Apochromat 63 × /1.4NA oil objective (Zeiss; Plan-Apochromat 63 × /1.4 Oil DIC M27). Samples stained with BG-dsDNA-Alexa488 were excited using a 488 nm laser. Emissions were collected through a 495–550 nm band-pass filter, 570 nm long-pass filter, and a 1.25 airy unit (~60 μm) pinhole onto a 32 GaAsP detector element. Pictures were obtained using ZEN software (Zeiss; black edition) and pixel size was set to ~40 nm in the x and y directions. SIM imaging was performed using a commercial microscope (NIKON, A1&SIM&STORM, Japan) with a SIM model equipped with a 100 × /1.49NA oil objective (Nikon; CFI Apo TIRF 100 × /1.49 Oil), and the 488 laser was selected.