Super-resolution photon-efficient imaging by nanometric double-helix point spread function localization of emitters (SPINDLE)

Abstract

Super-resolution imaging with photo-activatable or photoswitchable probes is a promising tool in biological applications to reveal previously unresolved intra-cellular details with visible light. This field benefits from developments in the areas of molecular probes, optical systems, and computational post-processing of the data. The joint design of optics and reconstruction processes using double-helix point spread functions (DH-PSF) provides high resolution three-dimensional (3D) imaging over a long depth-of-field. We demonstrate for the first time a method integrating a Fisher information efficient DH-PSF design, a surface relief optical phase mask, and an optimal 3D localization estimator. 3D super-resolution imaging using photo-switchable dyes reveals the 3D microtubule network in mammalian cells with localization precision approaching the information theoretical limit over a depth of 1.2 µm. Bewersdorf, "Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples," Nat. Methods 5(6), 527-529 (2008

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