Luminescent-reaction-enabled super-resolution imaging.

Abstract

By breaking the optical diffraction limit, super-resolution fluorescence microscopy has advanced our understanding of biological complexity under the framework of light-excited luminescence¹. The use of external light excitation remains a key factor that shapes the imaging capabilities and live-cell compatibility of fluorescence-based approaches². An alternative is the reaction-excited luminescence, such as electrochemiluminescence (ECL)³, chemiluminescence (CL)⁴ and bioluminescence (BL)⁵, providing a chemically defined toolbox for enabling different imaging merits, from ultrasensitive analysis^6,7 to biocompatible imaging^8,9. Despite its light-free excitation and high sensitivity, conventional luminescent-reaction-enabled imaging is fundamentally limited in spatiotemporal resolution owing to low photon budget^10,11. Here we develop a chemistry-based super-resolution imaging framework, luminescent-reaction-enabled super-resolution imaging via entropy-weighted correlation combined with deconvolution (RIED). As an experimental-computational concept, RIED introduces a spatiotemporal recording strategy to uncover specific luminescent-reaction-enabled imaging information content, which is efficiently collected and computed to achieve super resolution using a reconstruction strategy adapted to reaction-driven photon statistics. We achieve super-resolution ECL, CL and BL imaging of intracellular organelles, attaining approximately 100 nm resolution. This approach is used for highly sensitive imaging of surface proteins and 41-h ultralong-term continuous super-resolution live-cell imaging of mitochondrial transfer dynamics. Our work establishes an emerging class of chemistry-enabled, laser-free super-resolution microscopy with expanded biological imaging versatilities.