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- 摘要
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Successful optimization of reconstruction parameters in structured illumination microscopy – A practical guide
摘要： The impact of the different reconstruction parameters in super-resolution structured illumination microscopy (SIM) on image artifacts is carefully analyzed. These parameters comprise the Wiener filter parameter, an apodization function, zero-frequency suppression and modifications of the optical transfer function. A detailed investigation of the reconstructed image spectrum is concluded to be suitable for identifying artifacts. For this purpose, two samples, an artificial test slide and a more realistic biological system, were used to characterize the artifact classes and their correlation with the image spectra as well as the reconstruction parameters. In addition, a guideline for efficient parameter optimization is suggested and the implementation of the parameters in selected up-to-date processing packages (proprietary and open-source) is depicted.
关键词： Super resolution,Fluorescence microscopy,Structured illumination microscopy,Parameter estimation,Image reconstruction
更新于2025-09-23 15:23:52
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[IEEE OCEANS 2018 MTS/IEEE Charleston - Charleston, SC, USA (2018.10.22-2018.10.25)] OCEANS 2018 MTS/IEEE Charleston - Dynamic beam shaping exploiting orbital angular momentum for underwater structured light illumination
摘要： This work demonstrates dynamic optical beam generation using two different approaches: a Mach-Zehnder interferometer to generate three coherently coupled orbital angular momentum modes and a HOBBIT (high-order Bessel beams integrated in time) system which switches between spatial modes. By expanding the number of coherently coupled modes, a broad range of structured light patterns can be spatially and temporally controlled for structured illumination in underwater imaging applications.
关键词： underwater,orbital angular momentum,interference,structured illumination,coherent
更新于2025-09-23 15:22:29
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Magnification-invariant surface profiling technique for structured illumination imaging and microscopy
摘要： The structured illumination (SI) surface profiling method that is fast and has a depth-independent image magnification ratio is presented. Utilizing an electrically tunable lens (ETL) makes it possible to scan the object plane rapidly and motionlessly. The change in magnification ratio that generally occurs with the variation in the focal length of an objective lens is minimized by using a 4f relay system and placing the ETL at the confocal plane. The high reflection at the uncoated membrane surfaces of the ETL is bypassed by using polarization optics. It is also proposed that the fast response of the ETL can be fully exploited by switching the scanning sequence from that of a conventional SI imaging system; performing the depth scan before the pattern shift. It is shown that the scanning speed was increased up to 25 times. The principle and the related problems of SI profiling based on an ETL are fully analyzed, and the concept is confirmed experimentally. In the experiment, scanning depth variation of 35 mm could be achieved while keeping the magnification ratio variation below 0.03. The ability of 3D profiling is demonstrated by SI imaging a cone-shape 3D object and a face-shape plaster figure. In the fields requiring fast scanning, such as an intraoral scanner or biomedical imaging, the proposing SI imaging method can be fully utilized.
关键词： Tunable Lens,Surface Profiling,Intraoral scanner,Structured Illumination,Magnification invariant
更新于2025-09-23 15:21:01
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Structured-illumination Makyoh-topography: optimum grid position and its constraints
摘要： Complementing conventional Makyoh topography with structured illumination using a sparse square grid, the large-scale surface shape can be calculated with a deflectometry approach, while the sample’s morphology can still be imaged. However, the grid’s image must be sharp not to mask the Makyoh image of the sample morphology. In this paper, the instrumental conditions for the grid sharpness are established. The two main types of Makyoh set-ups (lens and mirror based) are analysed. It is shown that the lens-based set-ups allow the position of the grid to be sharp on the Makyoh image. However, for mirror-based set-ups this is not possible because of geometrical instrumental constraints. The calculations are corroborated with experiments.
关键词： Makyoh topography,surface defects,structured illumination,deflectometry,geometrical optics,flatness testing
更新于2025-09-23 15:21:01
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Polarization-Controlled Plasmonic Structured Illumination
摘要： Structured light in the subwavelength scale is important for a broad range of applications ranging from lithography to imaging. Of particular importance is the ability to dynamically shift the pattern of the fields, which has led to the development of structured illumination microscopy. Further extension of structured illumination to plasmonic systems has enabled imaging beyond diffraction limit. However, structured illumination usually requires complicated optical setups entailing moving mechanical parts. Here a polarization tunable structured plasmonic fields (SPF) is proposed and experimentally demonstrated. The SPF is formed by surface plasmon interference (SPI) generated by a fishbone-shaped metasurface on a thin gold film. Importantly, the SPF can be continuously shifted by merely varying the linear polarization state of an incident beam. The precise control of the fringes of structured illumination and elimination of mechanical control will have great potential in subdiffractional imaging for practical applications.
关键词： Plasmonic,metasurface,structured illumination,subdiffractional imaging
更新于2025-09-19 17:13:59
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<i>De Novo</i> -Designed Near-Infrared Nanoaggregates for Super-Resolution Monitoring of Lysosomes in Cells, in Whole Organoids, and <i>in Vivo</i>
摘要： As the cleaners of cells, lysosomes play an important role in circulating organic matter within cells, recovering damaged organelles, and removing waste via endocytosis. Because lysosome dysfunction is associated with various diseases—lysosomal storage diseases, inherited diseases, rheumatoid arthritis, and even shock—it is vital to monitor the movement of lysosomes in cells and in vivo. To that purpose, a method of optical imaging, super-resolution imaging technology (e.g., SIM and STORM), can overcome the limitations of traditional optical imaging and afford a range of possibilities for fluorescence imaging. However, the short wavelength excitation and easy photobleaching of super-resolution fluorescence probes somewhat problematize super-resolution imaging. As described herein, we designed a low-toxicity, photostable, near-infrared small molecule fluorescence probe HD-Br for use in the super-resolution imaging of lysosomes. The interaction of lysosomes and mitochondria was dynamically traced while using the probe’s properties to label the lysosomes. Because the probe has the optimal near-infrared excitation and emission wavelengths, liver organoid 3D imaging and Caenorhabditis elegans imaging were also performed. Altogether, our findings indicate valuable approaches and techniques for super-resolution 3D and in vivo imaging.
关键词： lysosome-targeted,3D organoids imaging,C. elegans imaging,structured illumination microscopy,nanoaggregates
更新于2025-09-11 14:15:04
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Time-Resolved Structured Illumination Microscopy for Phase Separation Dynamics of Water and 2-Butoxyethanol Mixtures: Interpretation of “Early Stage” Involving Micelle-Like Structures
摘要： Phase separation dynamics of a water/2-butoxyethanol (2BE) mixture was studied with newly developed time-resolved structured illumination microscopy (SIM). Interestingly, an employed hydrophobic fluorescent probe for SIM showed spectral shifts up to 500 ns after a laser-induced temperature jump, which suggests 2BE micellar-like aggregates become more hydrophobic at the initial stage of phase separation. This hydrophobic environment in 2BE aggregates, probably due to the ejection of water molecules, continued up to at least 10 μs. Time-resolved SIM and previously-reported light scattering data clearly showed that the size of a periodic structure remained constant (ca. 300 nm) from 3 to 10 μs, and then the growth of periodic structures having the self-similarity started. We think that the former and the latter processes correspond to “early stage” (concentration growth) and “late stage” (size growth), respectively, in phase separation dynamics. Here we suggest that, in the early stage, the entity to bear 2BE phase be water-poor 2BE aggregates, and the number density of these aggregates would simply increase in time.
关键词： phase separation dynamics,hydrophobic fluorescent probe,2-butoxyethanol,structured illumination microscopy,micellar-like aggregates
更新于2025-09-10 09:29:36
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Faster, sharper, and deeper: structured illumination microscopy for biological imaging
摘要： Structured illumination microscopy (SIM) allows rapid, super-resolution (SR) imaging in live specimens. We review recent technical advances in SR-SIM, with emphasis on imaging speed, resolution, and depth. Since its introduction decades ago, the technique has grown to offer myriad implementations, each with its own strengths and weaknesses. We discuss these, aiming to provide a practical guide for biologists and to highlight which approach is best suited to a given application.
关键词： Structured illumination microscopy,resolution,biological imaging,super-resolution imaging,imaging speed,depth
更新于2025-09-09 09:28:46
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[IEEE 2018 20th International Conference on Transparent Optical Networks (ICTON) - Bucharest (2018.7.1-2018.7.5)] 2018 20th International Conference on Transparent Optical Networks (ICTON) - Recent Advances in 3D Structured Illumination Microscopy
摘要： In structured illumination microscopy (SIM) the sample under investigation is illuminated using a structured illumination (SI) pattern. This SI pattern encodes high spatial frequencies of fine features within the sample, which usually are not transferred by the conventional three-dimensional (3D) optical transfer function (OTF) of the imaging system and fills the missing cone of frequencies in the OTF for better discrimination of the out-of- focus light. Thereby, SIM provides super-resolution (SR) performance beyond the diffraction limit and optical- sectioning (OS) capability with the use of data post-processing approaches. 3D structured patterns that include lateral and axial variations in the illumination have attracted more attention recently as they provide OS and SR enhancement in three dimensions. In this paper, we review recent implementations in generating a 3D SI pattern with tunable modulation frequency (independently of both the objective lens and the wavelength used) using a Fresnel biprism or a Wollaston prism. From the raw SIM images, final SIM images with simultaneous OS and SR are retrieved without artifacts due to coherent noise.
关键词： Fresnel biprism,Wollaston prism,structured illumination design,super-resolution,tunable frequency,three-dimensional microscopy
更新于2025-09-04 15:30:14
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Characterization of neurite dystrophy after trauma by high speed structured illumination microscopy and lattice light sheet microscopy
摘要： Background: Unbiased screening studies have repeatedly identified actin-related proteins as one of the families of proteins most influenced by neurotrauma. Nevertheless, the status quo model of cytoskeletal reorganization after neurotrauma excludes actin and incorporates only changes in microtubules and intermediate filaments. Actin is excluded in part because it is difficult to image with conventional techniques. However, recent innovations in fluorescent microscopy provide an opportunity to image the actin cytoskeleton at super-resolution resolution in living cells. This study applied these innovations to an in vitro model of neurotrauma. New method: New methods are introduced for traumatizing neurons before imaging them with high speed structured illumination microscopy or lattice light sheet microscopy. Also, methods for analyzing structured illumination microscopy images to quantify post-traumatic neurite dystrophy are presented. Results: Human induced pluripotent stem cell-derived neurons exhibited actin organization typical of immature neurons. Neurite dystrophy increased after trauma but was not influenced by jasplakinolide treatment. The F-actin content of dystrophies varied greatly from one dystrophy to another. Comparison with existing methods: In contrast to fixation dependent methods, these methods capture the evolution of the actin cytoskeleton over time in a living cell. In contrast to prior methods based on counting dystrophies, this quantification scheme parameterizes the severity of a given dystrophy as it evolves from a local swelling to an almost-perfect spheroid that threatens to transect the neurite. Conclusions: These methods can be used to investigate genetic factors and therapeutic interventions that modulate the course of neurite dystrophy after trauma.
关键词： Traumatic brain injury,Dystrophy,Structured illumination microscopy,Human induced pluripotent stem cell derived neurons,Lattice light sheet microscopy
更新于2025-09-04 15:30:14