Dual orca flash 4.0 digital cmos camera c13440

All data were acquired with a commercial Nikon Eclipse Ti2 inverted microscope (Nikon instruments, Tokyo, Japan), equipped with A1R confocal scanhead, N-SIM and N-STORM modules (Nikon instruments, Tokyo, Japan). The fully motorized automated microscope was controlled by the NIS Elements software (version 5.42.01, Laboratory Imaging s.r.o, Praha, Czech Republic). The system performs multicolor widefield, confocal, and single-molecule localization imaging thanks to a pE-4000 (CoolLED, Andover, UK) light source with 16 selectable LED wavelengths (Widefield microscopy) and a LU-NV laser unit (Nikon instruments, Tokyo, Japan) equipped with 5 laser lines (405 nm (23.1 mW), 440 nm (25.5 mW) 488 nm (79.1 mW), 561 nm (79 mW), and 647 nm (137 mW)) (Confocal, SIM, TIRF and dSTORM microscopy). The microscope is equipped with fluorescence filter sets for the optimal detection of the employed fluorochromes thanks to a double-layer turret allowing the combination of 5 cubes per layer. The upper turret is devoted to widefield and STORM imaging, with the lower one acting as an additional filter wheel. SIM images were instead collected through the lower layer by spatially calibrated filter cubes. Emitted light was collected by a CMOS camera (Dual ORCA Flash 4.0 Digital CMOS camera C13440, Hamamatsu, Tokio, Japan) set on a 16-bit scale detection modality (Widefield/SIM/TIRF/dSTORM).

All data were acquired with a commercial inverted Nikon Eclipse Ti2 microscope (Nikon instruments, Tokyo, Japan), equipped with an A1R confocal scanhead and N-SIM and N-STORM modules (Nikon instruments, Tokyo, Japan). The fully motorized automated microscope was controlled by the NIS Elements software (version 5.42.01). The system performed multicolor widefield, confocal, and single-molecule localization imaging thanks to (i) an LED light source (pE-4000, CoolLED, Andover, UK) with 16 selectable wavelengths for widefield microscopy; (ii) a laser unit (LU-NV, Nikon instruments, Tokyo, Japan) equipped with 5 laser lines (405 nm (23.1 mW), 440 nm (25.5 mW) 488 nm (79.1 mW), 561 nm (79 mW), 647 nm (137 mW)) for confocal microscopy, and (iii) a laser bench (L4Cc combiner, Oxxius S.A., Lannion, France) equipped with four high-power sources (405 nm (216 mW), 488 nm (240 mW), 561 nm (240 mW), 640 nm (360 mW)) and two acousto-optic modulators for single-molecule microscopy). Emitted light was filtered by a filter wheel (Optospin, Cairn Research Ltd., Faversham, Kent, UK) and then was collected by a CMOS camera (Dual ORCA Flash 4.0 Digital CMOS camera C13440, Hamamatsu, Tokyo, Japan) set on a 16-bit scale detection modality (Widefield/DNA PAINT).

Sample were imaged by a commercial inverted Nikon Eclipse Ti2 microscope (Nikon instruments, Tokyo, Japan) equipped with an A1R confocal scan-head and N-SIM and N-STORM modules (Nikon instruments, Tokyo, Japan) and controlled by NIS Elements software (version 5.42.01). For widefield microscopy, an LED light source (pE-4000, CoolLED, Andover, United Kingdom) with 16 selectable wavelengths for widefield microscopy was employed. The light source for confocal imaging was a laser unit (LU-NV, Nikon instruments, Tokyo, Japan) equipped with 5 laser lines (405 nm (23.1 mW), 440 nm (25.5 mW) 488 nm (79.1 mW), 561 nm (79 mW), 647 nm (137 mW)), while a laser bench (L4Cc combiner, Oxxius S.A., Lannion, France) equipped with four high-power sources (405nm (216 mW), 488 nm (240 mW), 561 nm (240 mW), 640 nm (360 mW)) was employed for single-molecule localization experiments. A filter wheel (Optospin, Cairn Research Ltd, Faversham, Kent, UK) was placed in front of a CMOS camera (Dual ORCA Flash 4.0 Digital CMOS camera C13440, Hamamatsu, Tokyo, Japan) to acquire 16-bit scaled images (Widefield/DNA PAINT).