Nanometer Resolution Imaging and Tracking of Single Fluorophores by Sequential Structured Illumination

Abstract

Localization of single fluorescent emitters is central for many physicochemical and biophysical measurements at the nanoscale and beyond ensemble averaging. Examples include single-molecule tracking and super-resolution imaging by single-molecule localization microscopy. Among the numerous localization methods available, MINFLUX constituted a breakthrough for achieving a ∼10-fold improvement in resolution over wide-field camera-based approaches, reaching the molecular scale at moderate photon counts. Since then, other related methods have been developed and a common framework has been established for single-molecule localization through sequential structured illumination. Now, fluorescence nanoscopy and tracking of single fluorophores with true nanometric resolution are sitting on solid conceptual and experimental ground. There is an ample scope of questions where they can be applied in the near future to gain new insight into the dynamics and photophysics of natural and synthetic systems at the nanometer scale.

Exploring the Synergies of Single‐Molecule Fluorescence and 2D Materials Coupled by DNA

“…[ 156 ] Furthermore, if high spatial and temporal resolution are simultaneously required, the so‐called MINFLUX technique [ 160 ] or its pulsed‐interleaved version, p‐MINFLUX, [ 10,158 ] are the methods of choice. [ 159 ] Understanding dynamic processes will gain in importance, resulting in a growing number of tracking applications. [ 159 ] Toward this end, unveiling fast conformational changes will be one step.…”

“…[ 159 ] Understanding dynamic processes will gain in importance, resulting in a growing number of tracking applications. [ 159 ] Toward this end, unveiling fast conformational changes will be one step. [ 160 ]…”

“…[159] Understanding dynamic processes will gain in importance, resulting in a growing number of tracking applications. [159] Toward this end, unveiling fast conformational changes will be one step. [160] While the proposed combination of different fields can be beneficial, it also introduces the challenges and drawbacks of each field.…”