Recent advancements in structured-illumination microscopy toward live-cell imaging

Microscopy ◽  
2015 ◽  
Vol 64 (4) ◽  
pp. 237-249 ◽  
Author(s):  
Yasuhiro Hirano ◽  
Atsushi Matsuda ◽  
Yasushi Hiraoka
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

scholarly journals Advances in High-Speed Structured Illumination Microscopy

2021 ◽  
Vol 9 ◽  
Author(s):  
Tianyu Zhao ◽  
Zhaojun Wang ◽  
Tongsheng Chen ◽  
Ming Lei ◽  
Baoli Yao ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
High Speed ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Light Power ◽  
Recent Developments ◽  
Super Resolution Microscopy

Super-resolution microscopy surpasses the diffraction limit to enable the observation of the fine details in sub-cellular structures and their dynamics in diverse biological processes within living cells. Structured illumination microscopy (SIM) uses a relatively low illumination light power compared with other super-resolution microscopies and has great potential to meet the demands of live-cell imaging. However, the imaging acquisition and reconstruction speeds limit its further applications. In this article, recent developments all targeted at improving the overall speed of SIM are reviewed. These comprise both hardware and software improvements, which include a reduction in the number of raw images, GPU acceleration, deep learning and the spatial domain reconstruction. We also discuss the application of these developments in live-cell imaging.

scholarly journals Motion artefact detection in structured illumination microscopy for live cell imaging

2016 ◽  
Vol 24 (19) ◽  
pp. 22121 ◽  
Author(s):  
Ronny Förster ◽  
Kai Wicker ◽  
Walter Müller ◽  
Aurélie Jost ◽  
Rainer Heintzmann
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Motion Artefact ◽  
Artefact Detection

Live Cell Imaging With Spatial Light Modulator-based Optical Sectioning Structured Illumination Microscopy

2014 ◽  
Vol 20 (S3) ◽  
pp. 388-389
Author(s):  
Zdeněk Švindrych ◽  
Pavel Křížek ◽  
Evgeny Smirnov ◽  
Martin Ovesný ◽  
Josef Borkovec ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Spatial Light Modulator ◽  
Cell Imaging ◽  
Live Cell ◽  
Structured Illumination ◽  
Optical Sectioning ◽  
Structured Illumination Microscopy ◽  
Light Modulator

scholarly journals Multi-color structured illumination microscopy for live cell imaging based on the enhanced image recombination transform algorithm

2021 ◽  
Vol 12 (6) ◽  
pp. 3474
Author(s):  
Tianyu Zhao ◽  
Huiwen Hao ◽  
Zhaojun Wang ◽  
Yansheng Liang ◽  
Kun Feng ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

Live Cell Super-Resolution Imaging with N-SIM

2012 ◽  
Vol 20 (4) ◽  
pp. 18-21
Author(s):  
Christopher B. O'Connell
Keyword(s):  
Light Microscope ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Specific Proteins ◽  
Resolution Imaging ◽  
Patterned Illumination

The ability to visualize the distributions of specific proteins with a light microscope and fluorescent probes is largely responsible for our current understanding of cellular structure. A major limitation of this approach arises from the blurring effects of diffraction, which decreases resolution and limits the ability to obtain information at the nanoscale. There has been a tremendous drive to develop optical and computational methods that improve the resolution of the light microscope, and structured illumination microscopy (SIM) is one solution. This method uses patterned illumination to double both lateral and axial resolution. Nikon's N-SIM is a commercial system that integrates the most desirable features of light microscopy, specific labeling of molecules, and live cell imaging, with structured illumination. This provides the ability to achieve super resolution suitable for a range of biological applications.

scholarly journals Highly photostable, reversibly photoswitchable fluorescent protein with high contrast ratio for live-cell superresolution microscopy

2016 ◽  
Vol 113 (37) ◽  
pp. 10364-10369 ◽  
Author(s):  
Xi Zhang ◽  
Mingshu Zhang ◽  
Dong Li ◽  
Wenting He ◽  
Jianxin Peng ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Fluorescent Protein ◽  
Contrast Ratio ◽  
Live Cell ◽  
Acquisition Time ◽  
Fluorescence Signal ◽  
Effective Quantum Yield ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

Two long-standing problems for superresolution (SR) fluorescence microscopy are high illumination intensity and long acquisition time, which significantly hamper its application for live-cell imaging. Reversibly photoswitchable fluorescent proteins (RSFPs) have made it possible to dramatically lower the illumination intensities in saturated depletion-based SR techniques, such as saturated depletion nonlinear structured illumination microscopy (NL-SIM) and reversible saturable optical fluorescence transition microscopy. The characteristics of RSFPs most critical for SR live-cell imaging include, first, the integrated fluorescence signal across each switching cycle, which depends upon the absorption cross-section, effective quantum yield, and characteristic switching time from the fluorescent “on” to “off” state; second, the fluorescence contrast ratio of on/off states; and third, the photostability under excitation and depletion. Up to now, the RSFPs of the Dronpa and rsEGFP (reversibly switchable EGFP) families have been exploited for SR imaging. However, their limited number of switching cycles, relatively low fluorescence signal, and poor contrast ratio under physiological conditions ultimately restrict their utility in time-lapse live-cell imaging and their ability to reach the desired resolution at a reasonable signal-to-noise ratio. Here, we present a truly monomeric RSFP, Skylan-NS, whose properties are optimized for the recently developed patterned activation NL-SIM, which enables low-intensity (∼100 W/cm2) live-cell SR imaging at ∼60-nm resolution at subsecond acquisition times for tens of time points over broad field of view.

scholarly journals Live-cell imaging of human spermatozoa using structured illumination microscopy

2018 ◽  
Vol 9 (12) ◽  
pp. 5939
Author(s):  
Ida S. Opstad ◽  
Daria A. Popova ◽  
Ganesh Acharya ◽  
Purusotam Basnet ◽  
Balpreet S. Ahluwalia
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Human Spermatozoa ◽  
Structured Illumination ◽  
Structured Illumination Microscopy
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