scholarly journals Cusp-artifacts in high order Superresolution Optical Fluctuation Imaging

2019 ◽  
Author(s):  
Xiyu Yi ◽  
Shimon Weiss
Keyword(s):  
Dynamic Range ◽  
Imaging Techniques ◽  
Photophysical Properties ◽  
Super Resolution ◽  
High Order ◽  
Fluorescence Probes ◽  
Structured Illumination ◽  
Odd Order ◽  
Even Order ◽  
High Order Cumulants

AbstractSuperresolution Optical Fluctuation Imaging (SOFI) offers a simple and affordable alternative to the more sophisticated (and expensive) super-resolution imaging techniques such as STED, PALM, STORM, structured illumination, and other derivative methods. In SOFI, the calculation of high order cumulants provides higher resolution but drastically expands the dynamic range of the resulting image. In this study, we have identified another type of artifact for high order SOFI cumulants, dubbed as ‘cusp artifacts.’ A series of realistic simulations are performed to study the cusp artifacts under the influences of various factors, including the blinking statistics, the spatial distribution of photophysical properties of the sample, the total number of frames processed per dataset, photobleaching, and noise. Experiments, simulations, and theory all show that high order cumulants and odd-order moments could suffer from cusp artifacts. These cusp artifacts also degrade the fidelity of bSOFI that has been proposed to solve the dynamic range expansion of image pixel intensities. Alternatively, cusp-artifacts could be altogether eliminated by utilizing even-order moments constructed directly or from cumulants for image reconstruction. Together with dynamic range compression, these approaches yield improved SOFI images. Our study provides new insight into the nature of high order SOFI images, outlines guidelines for developing and screening SOFI-optimized fluorescence probes, and suggests improved strategies for SOFI data acquisition.

scholarly journals Moments reconstruction and local dynamic range compression of high order Superresolution Optical Fluctuation Imaging

2018 ◽  
Author(s):  
Xiyu Yi ◽  
Sungho Son ◽  
Ryoko Ando ◽  
Atsushi Miyawaki ◽  
Shimon Weiss
Keyword(s):  
Dynamic Range ◽  
Imaging Techniques ◽  
Resolution Enhancement ◽  
Super Resolution ◽  
High Order ◽  
Live Cells ◽  
Local Dynamic ◽  
Actin Stress Fiber ◽  
Dynamic Range Compression ◽  
Even Order

Abstract:Super-resolution Optical Fluctuation Imaging (SOFI) offers a simple and affordable alternative to other super-resolution (SR) imaging techniques. The theoretical resolution enhancement of SOFI scales linearly with the cumulants’ order, while imaging conditions are less phototoxic to living samples as compared to other SR methods. High order SOFI could, therefore, be a method of choice for dynamic live cell imaging. However, due to cusp-artifacts and to dynamic range expansion of pixel intensities, this promise has not been materialized as of yet. Here we investigated and compared high order moments vs. high order cumulants SOFI reconstructions. We demonstrate that even-order moments reconstructions are intrinsically free of cusp artifacts, allowing for a subsequent deconvolution operation to be performed, hence enhancing the resolution even further. High order moments reconstructions performance was examined for various (simulated) conditions and applied to (experimental) imaging of QD labeled microtubules in fixed cells, and actin stress fiber dynamics in live cells.

Super-Resolution Imaging in Fluid Mechanics Using New Illumination Approaches

2020 ◽  
Vol 52 (1) ◽  
pp. 369-393
Author(s):  
Minami Yoda
Keyword(s):  
Fluid Mechanics ◽  
Total Internal Reflection ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Internal Reflection ◽  
Interfacial Flows ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Entire Volume ◽  
Resolution Imaging

Quantifying submillimeter flows using optical diagnostic techniques is often limited by a lack of spatial resolution and optical access. This review discusses two super-resolution imaging techniques, structured illumination microscopy and total internal reflection fluorescence or microscopy, which can visualize bulk and interfacial flows, respectively, at spatial resolutions below the classic diffraction limits. First, we discuss the theory and applications of structured illumination for optical sectioning, i.e., imaging a thin slice of a flow illuminated over its entire volume. Structured illumination can be used to visualize the interior of multiphase flows such as sprays by greatly reducing secondary scattering. Second, the theory underlying evanescent waves is introduced, followed by a review of how total internal reflection microscopy has been used to visualize interfacial flows over the last 15 years. Both techniques, which are starting to be used in fluid mechanics, could significantly improve quantitative imaging of microscale and macroscale flows.

An overview of structured illumination microscopy: recent advances and perspectives

2021 ◽  
Author(s):  
Krishnendu Samanta ◽  
Joby Joseph
Keyword(s):  
Spatial Frequency ◽  
Basic Principle ◽  
Imaging Techniques ◽  
Resolution Enhancement ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Structured Illumination ◽  
Post Processing ◽  
Structured Illumination Microscopy ◽  
Near Future

Abstract Structured illumination microscopy (SIM) is one of the most significant widefield super-resolution optical imaging techniques. The conventional SIM utilizes a sinusoidal structured pattern to excite the fluorescent sample; which eventually down-modulates higher spatial frequency sample information within the diffraction-limited passband of the microscopy system and provides around two-fold resolution enhancement over diffraction limit after suitable computational post-processing. Here we provide an overview of the basic principle, image reconstruction, technical development of the SIM technique. Nonetheless, in order to push the SIM resolution further towards the extreme nanoscale dimensions, several different approaches are launched apart from the conventional SIM. Among the various SIM methods, some of the important techniques e.g. TIRF, non-linear, plasmonic, speckle SIM etc. are discussed elaborately. Moreover, we highlight different implementations of SIM in various other imaging modalities to enhance their imaging performances with augmented capabilities. Finally, some future outlooks are mentioned which might develop fruitfully and pave the way for new discoveries in near future.

scholarly journals Super-Resolution Imaging of Tight and Adherens Junctions: Challenges and Open Questions

2020 ◽  
Vol 21 (3) ◽  
pp. 744 ◽  
Author(s):  
Hannes Gonschior ◽  
Volker Haucke ◽  
Martin Lehmann
Keyword(s):  
Single Molecule ◽  
Imaging Techniques ◽  
Rapid Development ◽  
Ion Homeostasis ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Molecular Architecture ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Resolution Imaging

The tight junction (TJ) and the adherens junction (AJ) bridge the paracellular cleft of epithelial and endothelial cells. In addition to their role as protective barriers against bacteria and their toxins they maintain ion homeostasis, cell polarity, and mechano-sensing. Their functional loss leads to pathological changes such as tissue inflammation, ion imbalance, and cancer. To better understand the consequences of such malfunctions, the junctional nanoarchitecture is of great importance since it remains so far largely unresolved, mainly because of major difficulties in dynamically imaging these structures at sufficient resolution and with molecular precision. The rapid development of super-resolution imaging techniques ranging from structured illumination microscopy (SIM), stimulated emission depletion (STED) microscopy, and single molecule localization microscopy (SMLM) has now enabled molecular imaging of biological specimens from cells to tissues with nanometer resolution. Here we summarize these techniques and their application to the dissection of the nanoscale molecular architecture of TJs and AJs. We propose that super-resolution imaging together with advances in genome engineering and functional analyses approaches will create a leap in our understanding of the composition, assembly, and function of TJs and AJs at the nanoscale and, thereby, enable a mechanistic understanding of their dysfunction in disease.

scholarly journals Structured illumination microscopy and its new developments

2016 ◽  
Vol 09 (03) ◽  
pp. 1630010 ◽  
Author(s):  
Jianling Chen ◽  
Caimin Qiu ◽  
Minghai You ◽  
Xiaogang Chen ◽  
Hongqin Yang ◽  
...  
Keyword(s):  
Optical Microscopy ◽  
Spatial Resolution ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Living Cells ◽  
The Other ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Recent Developments ◽  
Super Resolution Microscopy

Optical microscopy allows us to observe the biological structures and processes within living cells. However, the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light diffraction. Structured illumination microscopy (SIM), a type of new emerging super-resolution microscopy, doubles the spatial resolution by illuminating the specimen with a patterned light, and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy. In addition, SIM is easier to combine with the other imaging techniques to improve their imaging resolution, leading to the developments of diverse types of SIM. SIM has great potential to meet the various requirements of living cells imaging. Here, we review the recent developments of SIM and its combination with other imaging techniques.

scholarly journals Circumventing the optical diffraction limit with customized speckles

2020 ◽  
Author(s):  
Nicholas Bender ◽  
Mengyuan Sun ◽  
Hasan Yılmaz ◽  
Joerg Bewersdorf ◽  
Hui Cao
Keyword(s):  
Imaging Techniques ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Optical Diffraction ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Dynamic Speckle ◽  
Speckle Patterns ◽  
Super Resolution Microscopy ◽  
Fluorescence Photoswitching

Speckle patterns have been widely used in imaging techniques such as ghost imaging, dynamic speckle illumination microscopy, structured illumination microscopy, and photoacoustic fluctuation imaging. Recent advances in the ability to control the statistical properties of speckles has enabled the customization of speckle patterns for specific imaging applications. In this work, we design and create special speckle patterns for parallelized nonlinear pattern-illumination microscopy based on fluorescence photoswitching. We present a proof-of-principle experimental demonstration where we obtain a spatial resolution three times higher than the diffraction limit of the illumination optics in our setup. Furthermore, we show that tailored speckles vastly outperform standard speckles. Our work establishes that customized speckles are a potent tool in parallelized super-resolution microscopy.

scholarly journals Super-Resolved Fluorescence Imaging of Peripheral Nerve  

2021 ◽  
Author(s):  
Iván Coto Hernández ◽  
Suresh Mohan ◽  
Nate Jowett
Keyword(s):  
Peripheral Nerve ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Biomedical Science ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Unmyelinated Axons ◽  
Histopathologic Evaluation ◽  
Resolution Imaging

Abstract Traditional histopathologic evaluation of peripheral nerve employs brightfield microscopy with diffraction limited resolution of ~ 250 nm. Though electron microscopy yields nanoscale resolution of the nervous system, it is resource-intensive and incompatible with life. Super-resolution microscopy (SRM) comprises a set of imaging techniques permitting unprecedented resolution of fluorescent objects using visible light. The advent of SRM has transformed biomedical science in establishing non-toxic means for investigation of nanoscale cellular structures. Herein, sciatic nerve sections from GFP-variant expressing mice, and regenerating human nerve from cross-facial autografts labelled with a myelin-specific fluorescent dye were imaged by super-resolution radial fluctuation microscopy, stimulated emission depletion microscopy, and structured illumination microscopy. Super-resolution imaging of axial cryosections of murine sciatic nerves demonstrated robust visualization of myelinated and unmyelinated axons. Super-resolution imaging of axial cryosections of human cross-facial nerve grafts demonstrated enhanced resolution of small-calibre thinly-myelinated regenerating motor axons. The utility of SRM in imaging of mammalian cranial and peripheral nerves is demonstrated. The increase in contrast and structural clarity achievable with super-resolution techniques enables visualization of unmyelinated axons, regenerating axons, cytoskeleton ultrastructure, and neuronal appendages using light microscopes.

scholarly journals High-Order Interpolation Algorithms for Charge Conservation in Particle-in-Cell Simulations

2013 ◽  
Vol 13 (4) ◽  
pp. 1134-1150 ◽  
Author(s):  
Jinqing Yu ◽  
Xiaolin Jin ◽  
Weimin Zhou ◽  
Bin Li ◽  
Yuqiu Gu
Keyword(s):  
Particle Trajectory ◽  
High Order ◽  
Third Order ◽  
Charge Conservation ◽  
Particle In Cell ◽  
Interpolation Algorithms ◽  
Odd Order ◽  
Even Order ◽  
Pic Code ◽  
High Order Interpolation

AbstractHigh-order interpolation algorithms for charge conservation in Particle-in-Cell (PIC) simulations are presented. The methods are valid for the case that a particle trajectory is a zigzag line. The second-order and third-order algorithms which can be applied to any even-order and odd-order are discussed in this paper, respectively. Several test simulations are performed to demonstrate their validity in two-dimensional PIC code. Compared with the simulation results of one-order, high-order algorithms have advantages in computation precision and enlarging the grid scales which reduces the CPU time.

scholarly journals A Review of Super-Resolution Imaging through Optical High-Order Interference [Invited]

2019 ◽  
Vol 9 (6) ◽  
pp. 1166 ◽  
Author(s):  
Peilong Hong ◽  
Guoquan Zhang
Keyword(s):  
Optical Imaging ◽  
Single Photon ◽  
Near Field ◽  
Super Resolution ◽  
High Order ◽  
Structured Illumination ◽  
Wave Nature ◽  
Light Matter Interaction ◽  
Resolution Imaging ◽  
Rayleigh Limit

Resolution is crucially important for optical imaging, which defines the smallest spatial feature of object that can be delivered by light wave. However, due to the wave nature of light, optical imaging is of limited resolution, widely known as Rayleigh limit or Abbe limit. Nevertheless, this limit can be overcome by considering the loopholes in the derivation of the Rayleigh limit, such as light–matter interaction, structured illumination, and near-field interference. In contrast to the conventional single-photon interference, multi-photon amplitudes responsible for optical high-order interference could be designed to possess a reduced effective wavelength, enabling the breakthrough of the Rayleigh limit. In this review, we will present recently developed super-resolution imaging schemes based on optical high-order interference, and discuss future perspectives.

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