SSBD:database

Detail of Fig6_SDSRM

Shinichi Hayashi, Yasushi Okada (2015) Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics., Molecular biology of the cell, Volume 26, Number 9, pp. 1743-51

Published in 2015 May 1 (Electronic publication in Feb. 25, 2015, midnight )

• doi: 10.1091/mbc.E14-08-1287
• pmid: 25717185
  

(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.

• Animals
• Cell Line
• Endosomes/ultrastructure
• Microscopy, Confocal
• Microtubules/ultrastructure
• Mitochondria/ultrastructure
• Mitochondrial Membranes/ultrastructure
• Optical Imaging/instrumentation[major]
• Potoroidae
• Single-Cell Analysis/instrumentation[major]
• Transport Vesicles/ultrastructure