Nanosecond time-resolved fluorescence nanoscopy: 3D sub-10 nm imaging and super-resolved fluorescence lifetime imaging

Super-resolution microscopy, or more accurately fluorescence nanoscopy techniques have revolutionized fluorescence based imaging reaching resolutions in the nanometer range, well beyond the diffraction limit of light. Still, current technologies are not able to provide sub-10 nm resolution in all three dimensions. This project aims to develop a new fluorescence nanoscopy methodology capable with nanosecond time-resolution, so as to determine accurately fluorescence excited state lifetime of each super-resolved region.

By combining single-molecule localization nanoscopy (DNA-PAINT) for lateral resolution and time-resolved energy transfer to a metal surface (MIET) for axial resolution, imaging with a resolution of 1 to 6 nm in all three dimensions will be achieved. In addition, the new nanoscope will be capable of delivering for the first time super-resolved fluorescence lifetime imaging (SR-FLIM) and super-resolved Förster resonant energy transfer images (SR-FRET).