Detail of Fig8_SDSRM
Project
Title
Time-lapse SDSRM images of the dynamics of the outer membrane of mitochondria stained with TOMM20-mEmelard in MDCK cells
Description
NA
Release, Updated
2017-10-03,
2018-11-15
2018-11-15
License
CC BY-NC-SA
Kind
Image data
based on Experiment
File Formats
Data size
26.0 MB
Protein names
TOMM20
Protein tags
mEmelard
Datatype
organelle dynamics
Molecular Function (MF)
Biological Process (BP)
-
Cellular Component (CC)
organelle
(
GO:0043226
)
Biological Imaging Method
XYZ Scale
XY: 100 nanometer per slice, Z: NA
T scale
0.1 second for each time interval
Image Acquisition
Experiment type
TimeLapse
Microscope type
Other
Acquisition mode
StructuredIllumination
Contrast method
Fluorescence
Microscope model
Custom made microscope system based on IX81 (Olympus) with a disk-scannning unit IX2-DSU (Olympus), custom-made disk (DU-DSR-SP; Olympus) and laser illumination optics (Sapphire488HP; Coherent)
Detector model
ORCA FLASH 4.0 v2
Objective model
Olympus UPLSAPO60XO 60x/1.3 O
Filter set
Summary of Methods
See details in Hayashi and Okada (2015) Mol Biol Cell, 26(9): 1743-1751.
Related paper(s)
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.(MeSH Terms)
Contact
Yasushi Okada
, RIKEN
, Quantitative Biology Center
, Laboratory for Cell Polarity Regulation
Contributors
Shinichi Hayashi, Yasushi Okada
OMERO Dataset
OMERO Project
Source