Nanosilver-Enhanced Far-Field Fluorescence Fluctuations for Super-Resolution Microscopy

Fluctuation-based super-resolution microscopy enhances image resolution using signal fluctuations, yet the inherent fluctuations of fluorophores limit its spatiotemporal resolution. In this work, we reveal a far-field enhancement (FFE) effect via a nanosilver film that significantly boosts fluorescence fluctuations of fluorophores positioned up to 10 mu m away. The FFE effect arises from the interference of scattered light from the nanosilver film and photothermal-induced refractive index changes in the imaging medium, which create periodic auxiliary illumination on the sample. This phenomenon enabled the development of far-field enhanced super-resolution microscopy (FFE-SRM), a technique compatible with commonly used fluorophores. FFE-SRM improves temporal resolution up to 10-fold and enhances spatial resolution by about 2-fold over various SRM methods, including stochastic optical fluctuation imaging, super-resolution radial fluctuation, mean-shift super-resolution, and direct stochastic optical reconstruction microscopy. We demonstrated the potential of FFE-SRM by revealing mitochondrial dynamics in live-cell imaging, advancing super-resolution imaging, and cellular process exploration.