Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy

TLDR

The sample is illuminated with a series of excitation light patterns that encode normally inaccessible high‑resolution information into the observed image, and the recorded images are linearly processed to reconstruct a twice‑resolution image. The method achieves a lateral resolution twice the classical diffraction limit without discarding any emission light, producing images of strikingly increased clarity compared to conventional and confocal microscopes.

Abstract

Lateral resolution that exceeds the classical diffraction limit by a factor of two is achieved by using spatially structured illumination in a wide‐field fluorescence microscope. The sample is illuminated with a series of excitation light patterns, which cause normally inaccessible high‐resolution information to be encoded into the observed image. The recorded images are linearly processed to extract the new information and produce a reconstruction with twice the normal resolution. Unlike confocal microscopy, the resolution improvement is achieved with no need to discard any of the emission light. The method produces images of strikingly increased clarity compared to both conventional and confocal microscopes.