title: Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.
abstract: Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging.
authors.affiliation: Products Development Department 6, R&D Division, Olympus Corporation, 2951 Ishikawa-cho, Hachioji, Tokyo 192-8507, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Laboratory for Cell Polarity Regulation, Quantitative Biology Center, RIKEN, Furuedai 6-2-3, Suita, Osaka 565-0874, Japan y.okada@riken.jp.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okada
mesh_terms: D000818:Animals; D002460:Cell Line; D011992:Endosomes / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008870:Microtubules / Q000648:ultrastructure; D008928:Mitochondria / Q000648:ultrastructure; D051336:Mitochondrial Membranes / Q000648:ultrastructure; D061848:Optical Imaging / Q000295:instrumentation*; D048328:Potoroidae; D059010:Single-Cell Analysis / Q000295:instrumentation*; D022161:Transport Vesicles / Q000648:ultrastructure