Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging

ACS Photonics ◽  
2021 ◽  
Author(s):  
Alice Sandmeyer ◽  
Mario Lachetta ◽  
Hauke Sandmeyer ◽  
Wolfgang Hübner ◽  
Thomas Huser ◽  
...  
Keyword(s):  
Cost Effective ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

scholarly journals Extending resolution of structured illumination microscopy with sparse deconvolution

2021 ◽  
Author(s):  
Weisong Zhao ◽  
Shiqun Zhao ◽  
Liuju Li ◽  
Xiaoshuai Huang ◽  
Shijia Xing ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Super Resolution ◽  
Live Cell ◽  
Frame Rate ◽  
Live Cells ◽  
Photon Flux ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Spatiotemporal Resolution

Abstract The spatial resolutions of live-cell super-resolution microscopes are limited by the maximum collected photon flux. Taking advantage of a priori knowledge of the sparsity and continuity of biological structures, we develop a deconvolution algorithm that further extends the resolution of super-resolution microscopes under the same photon budgets by nearly twofold. As a result, sparse structured illumination microscopy (Sparse-SIM) achieves ~60 nm resolution at a 564 Hz frame rate, allowing it to resolve intricate structural intermediates, including small vesicular fusion pores, ring-shaped nuclear pores formed by different nucleoporins, and relative movements between the inner and outer membranes of mitochondria in live cells. Likewise, sparse deconvolution can be used to increase the three-dimensional resolution and contrast of spinning-disc confocal-based SIM (SD-SIM), and operates under conditions with the insufficient signal-to-noise-ratio, all of which allows routine four-color, three-dimensional, ~90 nm resolution live-cell super-resolution imaging. Overall, sparse deconvolution may be a general tool to push the spatiotemporal resolution limits of live-cell fluorescence 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 Quantification of cristae architecture reveals time-dependent characteristics of individual mitochondria

2020 ◽  
Vol 3 (7) ◽  
pp. e201900620
Author(s):  
Mayuko Segawa ◽  
Dane M Wolf ◽  
Nan W Hultgren ◽  
David S Williams ◽  
Alexander M van der Bliek ◽  
...  
Keyword(s):  
Machine Learning ◽  
Mitochondrial Fission ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Fusion Event ◽  
New Strategy ◽  
Function Relationship ◽  
Relationship Of ◽  
Standard Techniques

Recent breakthroughs in live-cell imaging have enabled visualization of cristae, making it feasible to investigate the structure–function relationship of cristae in real time. However, quantifying live-cell images of cristae in an unbiased way remains challenging. Here, we present a novel, semi-automated approach to quantify cristae, using the machine-learning Trainable Weka Segmentation tool. Compared with standard techniques, our approach not only avoids the bias associated with manual thresholding but more efficiently segments cristae from Airyscan and structured illumination microscopy images. Using a cardiolipin-deficient cell line, as well as FCCP, we show that our approach is sufficiently sensitive to detect perturbations in cristae density, size, and shape. This approach, moreover, reveals that cristae are not uniformly distributed within the mitochondrion, and sites of mitochondrial fission are localized to areas of decreased cristae density. After a fusion event, individual cristae from the two mitochondria, at the site of fusion, merge into one object with distinct architectural values. Overall, our study shows that machine learning represents a compelling new strategy for quantifying cristae in living cells.

Evaluating resolution in live cell structured illumination microscopy

2019 ◽  
Author(s):  
Jakub Pospisil ◽  
Karel Fliegel ◽  
Jan Švihlík ◽  
Miloš Klíma
Keyword(s):  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

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 Tiled Reconstruction Improves Structured Illumination Microscopy

2020 ◽  
Author(s):  
David P. Hoffman ◽  
Eric Betzig
Keyword(s):  
Super Resolution ◽  
Reconstruction Algorithm ◽  
Live Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Resolution Imaging

AbstractStructured illumination microscopy (SIM) is widely used for fast, long-term, live-cell super-resolution imaging. However, SIM images can contain substantial artifacts if the sample does not conform to the underlying assumptions of the reconstruction algorithm. Here we describe a simple, easy to implement, process that can be combined with any reconstruction algorithm to alleviate many common SIM reconstruction artifacts and briefly discuss possible extensions.

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
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