Sprecher
Beschreibung
Super-resolution microscopy has transformed biological imaging by enabling visualization of sub-cellular structures at nanometer scales beyond the diffraction limit, while also opening new possibilities for single-molecule sensing, nanomaterials characterization, and the study of molecular interactions with unprecedented spatial precision. Single-molecule localization microscopy (SMLM) techniques, such as DNA-PAINT, routinely achieve lateral resolution of approximately 10 nm. Sequential imaging approaches, such as Exchange-PAINT and Resolution Enhancement by Sequential Imaging (RESI), further improve spatial precision to the sub-nanometer scale by capturing targets over multiple imaging rounds. While these methods achieve remarkable accuracy, they come at the cost of long imaging cycles and reduced experimental practicality. Simultaneous multiplexed imaging presents a promising alternative, but current implementations are hindered by a limited fluorophore palette, spectral crosstalk, chromatic aberrations, and the need for elaborate alignment and calibration, typically restricting them to three colors. To overcome these limitations and push the boundaries of both spatial resolution and multiplexing, we introduce a novel wide-field SMLM technique that integrates fluorescence lifetime imaging microscopy (FLIM). By encoding different targets with distinct fluorescence lifetimes, our method enables simultaneous multi-target imaging in the same field of view. Benchmarked on DNA origami, our technique offers a scalable, high-resolution alternative for structural biology and cell imaging applications.
