High-speed 10-plex DNA-PAINT with a larger sequence repertoire

Abstract DNA-Points Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) enables multiplexed super-resolution imaging of biological samples. We expand the repertoire of speed-optimized DNA sequences for DNA-PAINT imaging to facilitate the molecular-resolution imaging of up to ten targets in a sequential manner. These imager sequences showcase a range of binding kinetics, providing the flexibility to select optimal sequences for specific applications. We demonstrate the application of twelve speed-optimized imager sequences for multiplexed super-resolution imaging of the microtubular network and Nuclear Lamins in the cellular context. We anticipate that the ability to image as many as ten targets using the simple Exchange-PAINT strategy at accelerated speed will drive its further adoption for diverse cellular imaging applications.