High-speed super-resolution imaging of rotationally symmetric structures using SPEED microscopy and 2D-to-3D transformation

Li, Yichen; Tingey, Mark; Ruba, Andrew; Yang, Weidong

doi:10.1038/s41596-020-00440-x

Cited by 19 publications

(38 citation statements)

References 91 publications

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“…The fast advancement in super resolution imaging and digital image processing/analysis definitely will elevate our understanding towards the chromatin architecture and chromatin dynamics. The recent emerging super resolution imaging techniques would enable the dynamic visualization of the chromatin structure at a video rate (Li et al, 2021; Wu et al, 2021), this will help to integrate the chromatin motion with the conformational change of the chromatin (Lakadamyali & Cosma, 2020). In addition, the utilization of artificial intelligence in image processing has revolutionized the digital image processing and analysis.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, it is found that active RNA polymerase II (RNAPII) globally constrains chromatin movements, and RNAPII inhibition or its rapid depletion released the chromatin constraints and increased chromatin dynamics (Nagashima et al, 2019) (Figure 3c). ture at a video rate (Li et al, 2021;Wu et al, 2021), this will help to integrate the chromatin motion with the conformational change of the chromatin (Lakadamyali & Cosma, 2020). In addition, the utilization of artificial intelligence in image processing has revolutionized the digital image processing and analysis.…”

Section: Chromatin Motion and Transcriptionmentioning

confidence: 99%

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A guide for single‐particle chromatin tracking in live cell nuclei

Zhang

Seitz

Chang

et al. 2022

Cell Biology International

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The emergence of labeling strategies and live cell imaging methods enables the imaging of chromatin in living cells at single digit nanometer resolution as well as milliseconds temporal resolution. These technical breakthroughs revolutionize our understanding of chromatin structure, dynamics and functions. Single molecule tracking algorithms are usually preferred to quantify the movement of these intranucleus elements to interpret the spatiotemporal evolution of the chromatin. In this review, we will first summarize the fluorescent labeling strategy of chromatin in live cells which will be followed by a systematic comparison of live cell imaging instrumentation. With the proper microscope, we will discuss the image analysis pipelines to extract the biophysical properties of the chromatin. Finally, we expect to give practical suggestions to broad biologists on how to select methods and link to the model properly according to different investigation purposes.

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Section: Discussionmentioning

confidence: 99%

Section: Chromatin Motion and Transcriptionmentioning

confidence: 99%

A guide for single‐particle chromatin tracking in live cell nuclei

Zhang

Seitz

Chang

et al. 2022

Cell Biology International

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“…In another study focused on the dynamics of mRNA export in the absence of targeted Basket Nups, Li and colleagues employed AID strategies to observe the alterations in export behavior or mRNA via SPEED microscopy [ 129 ]. SPEED microscopy is a super-resolution technique permitting a spatial localization precision during in vivo imaging of approximately 10 nm and a temporal resolution up to 0.4 ms that has been employed to great effect to track macromolecules as they move through the NPC or primary cilium of the cell [ 144 , 163–167 ]. In addition to deriving dynamic information regarding the docking and export behavior of macromolecules, SPEED microscopy makes effective use of a 2D-to-3D transformation algorithm.…”

Section: Basket Nups and Their Impact On Nuclear Export Of Mrnamentioning

confidence: 99%

Spelling out the roles of individual nucleoporins in nuclear export of mRNA

Tingey

et al. 2022

Nucleus

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The Nuclear Pore Complex (NPC) represents a critical passage through the nuclear envelope for nuclear import and export that impacts nearly every cellular process at some level. Recent technological advances in the form of Auxin Inducible Degron (AID) strategies and Single-Point Edge-Excitation sub-Diffraction (SPEED) microscopy have enabled us to provide new insight into the distinct functions and roles of nuclear basket nucleoporins (Nups) upon nuclear docking and export for mRNAs. In this paper, we provide a review of our recent findings as well as an assessment of new techniques, updated models, and future perspectives in the studies of mRNA’s nuclear export.

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“…After data collection, post-localization transform algorithms or/and simulations will be applied to obtain super-resolution structural and dynamic information. The complete protocol includes cell culture and sample preparation, SPEED microscopy imaging, data analysis, and validation through simulation, which takes approximately nine days to complete [67].…”

Section: Limitations Of Speed Microscopymentioning

confidence: 99%

Nuclear Import of Adeno-Associated Viruses Imaged by High-Speed Single-Molecule Microscopy

Junod

Saredy

Yang

2021

Viruses

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Understanding the detailed nuclear import kinetics of adeno-associated virus (AAV) through the nuclear pore complex (NPC) is essential for the application of AAV capsids as a nuclear delivery instrument as well as a target for drug development. However, a comprehensive understanding of AAV transport through the sub-micrometer NPCs in live cells calls for new techniques that can conquer the limitations of conventional fluorescence microscopy and electron microscopy. With recent technical advances in single-molecule fluorescence microscopy, we are now able to image the entire nuclear import process of AAV particles and also quantify the transport dynamics of viral particles through the NPCs in live human cells. In this review, we initially evaluate the necessity of single-molecule live-cell microscopy in the study of nuclear import for AAV particles. Then, we detail the application of high-speed single-point edge-excitation sub-diffraction (SPEED) microscopy in tracking the entire process of nuclear import for AAV particles. Finally, we summarize the major findings for AAV nuclear import by using SPEED microscopy.

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High-speed super-resolution imaging of rotationally symmetric structures using SPEED microscopy and 2D-to-3D transformation

Cited by 19 publications

References 91 publications

A guide for single‐particle chromatin tracking in live cell nuclei

A guide for single‐particle chromatin tracking in live cell nuclei

Spelling out the roles of individual nucleoporins in nuclear export of mRNA

Nuclear Import of Adeno-Associated Viruses Imaged by High-Speed Single-Molecule Microscopy

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