Nonlinear structured-illumination microscopy with a photoswitchable protein reveals cellular structures at 50-nm resolution.

ABSTRACT: Using ultralow light intensities that are well suited for investigating biological samples, we demonstrate whole-cell superresolution imaging by nonlinear structured-illumination microscopy. Structured-illumination microscopy can increase the spatial resolution of a wide-field light microscope by a factor of two, with greater resolution extension possible if the emission rate of the sample responds nonlinearly to the illumination intensity. Saturating the fluorophore excited state is one such nonlinear response, and a realization of this idea, saturated structured-illumination microscopy, has achieved approximately 50-nm resolution on dye-filled polystyrene beads. Unfortunately, because saturation requires extremely high light intensities that are likely to accelerate photobleaching and da

SUBMITTER: Rego EH 

PROVIDER: S-EPMC3271870 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature