Et600 50m

DNA-PAINT imaging was carried out on an inverted microscope (Nikon Instruments, Eclipse Ti2) with the Perfect Focus System, applying an objective-type total internal reflection fluorescence (TIRF) configuration equipped with an oil-immersion objective (Nikon Instruments, Apo SR TIRF 100×, NA 1.49, oil). A 488-nm (200 mW, Toptica iBeam smart) or 561-nm laser (Coherent Sapphire, 200 mW) was used for excitation and was coupled into a single-mode fiber. The laser beam was passed through cleanup filters (Chroma Technology, ZET561/10) and coupled into the microscope objective using a beam splitter (Chroma Technology, ZT561rdc). Fluorescence light was spectrally filtered with an emission filter (Chroma Technology, ET600/50 m) and imaged with a scientific complementary metal-oxide semiconductor camera (Andor, Zyla 4.2plus) without further magnification, resulting in an effective pixel size of 130 nm after 2 × 2 binning. The camera readout sensitivity was set to 16-bit, and readout bandwidth to 540 MHz. Three-dimensional imaging was performed using a cylindrical lens (Nikon Instruments, N-STORM) in the detection path.

Fluorescence imaging was carried out on an inverted microscope (Nikon Instruments, Eclipse Ti2) with the Perfect Focus System, applying an objective-type TIRF configuration with an oil-immersion objective (Nikon Instruments, Apo SR TIRF 100×, NA 1.49, Oil). A 561 nm and 640 nm (MPB Communications Inc, 2W, DPSS-system) laser were used for excitation. The laser beam was passed through clean-up filters (Chroma Technology, ZET561/10, ZET642/20x) and coupled into the microscope objective using a beam splitter (Chroma Technology, ZT561rdc, ZT647rdc). Fluorescence light was spectrally filtered with an emission filter (Chroma Technology, ET600/50m and ET575lp, ET705/72m and ET665lp) and imaged on a sCMOS camera (Andor, Zyla 4.2 Plus) without further magnification, resulting in an effective pixel size of 130 nm (after 2 × 2 binning).