FCS measurements were performed on a Nikon A1 confocal microscope (Nikon Instrument, Tokyo, Japan) with a 40 × objective NA = 1, 25 (Nikon Instrument, Tokyo, Japan). Excitation Argon LASER 488 nm (Coherent, USA) was used to scan samples in confocal illumination. To spectrally select the excitation from the emitted light, we used a bandpass dichroic mirror (405/488 nm). Fluorescence photons were collected on a photodiode and images were reconstructed with NIS software (Nikon Instruments, Tokyo, Japan).
To perform FCS, the CLSM was adapted with a TCSPC module (PicoQuant, Germany), and a pulsed laser at 488 nm (LDH-D-C 488, PicoQuant), a TSCPC unit, and a single molecule counting detector (SPCM CD 3516H, Excelitas Technologies Corp., USA). To drive this photon counting system, we used SymphoTime software (PicoQuant, Germany) in FCS mode. A high pass filter (BLP-488R, SEMROCK, USA) was set before the detector.
To perform FCS, the CLSM was adapted with a TCSPC module (PicoQuant, Germany), and a pulsed laser at 488 nm (LDH-D-C 488, PicoQuant), a TSCPC unit, and a single molecule counting detector (SPCM CD 3516H, Excelitas Technologies Corp., USA). To drive this photon counting system, we used SymphoTime software (PicoQuant, Germany) in FCS mode. A high pass filter (BLP-488R, SEMROCK, USA) was set before the detector.