scholarly journals Superaufgelöste Mikroskopie: Pilze unter Beobachtung

BIOspektrum ◽  
2021 ◽  
Vol 27 (4) ◽  
pp. 380-382
Author(s):  
Sebastian Sputh ◽  
Sabine Panzer ◽  
Christian Stigloher ◽  
Ulrich Terpitz
Keyword(s):  
Electron Microscopy ◽  
Membrane Protein ◽  
Fluorescence Imaging ◽  
Filamentous Fungus ◽  
Light And Electron Microscopy ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Fusarium Fujikuroi ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

AbstractThe diffraction limit of light confines fluorescence imaging of subcellular structures in fungi. Different super-resolution methods are available for the analysis of fungi that we briefly discuss. We exploit the filamentous fungus Fusarium fujikuroi expressing a YFP-labeled membrane protein showing the benefit of correlative light- and electron microscopy (CLEM), that combines structured illumination microscopy (SIM) and scanning election microscopy (SEM).

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.

DEEP-UV CONFOCAL FLUORESCENCE IMAGING AND SUPER-RESOLUTION OPTICAL MICROSCOPY OF BIOLOGICAL SAMPLES

2012 ◽  
Vol 05 (04) ◽  
pp. 1250025 ◽  
Author(s):  
TREVOR A. SMITH ◽  
LIISA M. HIRVONEN ◽  
CRAIG N. LINCOLN ◽  
XIAOTAO HAO
Keyword(s):  
Fluorescence Imaging ◽  
Biological Samples ◽  
Super Resolution ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Diffractive Imaging ◽  
Wide Range ◽  
Confocal Fluorescence ◽  
Deep Uv ◽  
Confocal Fluorescence Imaging

A wide range of techniques has been developed to image biological samples at high spatial and temporal resolution. In this paper, we report recent results from deep-UV confocal fluorescence microscopy to image inherent emission from fluorophores such as tryptophan, and structured illumination microscopy (SIM) of biological materials. One motivation for developing deep-UV fluorescence imaging and SIM is to provide methods to complement our measurements in the emerging field of X-ray coherent diffractive imaging.

scholarly journals Chlamydomonas FAP70 is a component of the previously uncharacterized ciliary central apparatus projection C2a

2021 ◽  
Author(s):  
Yuqing Hou ◽  
Lei Zhao ◽  
Tomohiro Kubo ◽  
Xi Cheng ◽  
Nathan McNeill ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mass Spectrometry ◽  
Chlamydomonas Reinhardtii ◽  
Model Organism ◽  
Super Resolution ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Cryo Electron Microscopy ◽  
N Terminus ◽  
Central Apparatus

Cilia are essential organelles required for cell signaling and motility. Nearly all motile cilia have a “9+2” axoneme composed of 9 outer doublet microtubules plus 2 central microtubules; the central microtubules together with their projections is termed the central apparatus (CA). In Chlamydomonas reinhardtii, a model organism for studying cilia, 30 proteins are known CA components, and ∼36 more are predicted to be CA proteins. Among the candidate CA proteins is the highly conserved FAP70, which also has been reported to be associated with the doublet microtubules. Here we determined by super-resolution structured illumination microscopy that FAP70 is located exclusively in the CA, and show by cryo-electron microscopy that its N-terminus is located at the base of the CA's C2a projection. We also found that fap70-1 mutant axonemes lack most of the C2a projection. Mass spectrometry revealed that fap70-1 axonemes lack not only FAP70 but two other conserved candidate CA proteins, FAP65 and FAP147. Finally, FAP65 and FAP147 co-immunoprecipitated with HA-tagged FAP70. Taken together, these data identify FAP70, FAP65, and FAP147 as the first defining components of the C2a projection.

scholarly journals Recent Progress in the Correlative Structured Illumination Microscopy

Chemosensors ◽  
2021 ◽  
Vol 9 (12) ◽  
pp. 364
Author(s):  
Meiting Wang ◽  
Jiajie Chen ◽  
Lei Wang ◽  
Xiaomin Zheng ◽  
Jie Zhou ◽  
...  
Keyword(s):  
High Frequency ◽  
Optical Transmission ◽  
Imaging Technique ◽  
Super Resolution ◽  
Diffraction Limit ◽  
Optical Diffraction ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Recent Developments ◽  
Resolution Imaging

The super-resolution imaging technique of structured illumination microscopy (SIM) enables the mixing of high-frequency information into the optical transmission domain via light-source modulation, thus breaking the optical diffraction limit. Correlative SIM, which combines other techniques with SIM, offers more versatility or higher imaging resolution than traditional SIM. In this review, we first briefly introduce the imaging mechanism and development trends of conventional SIM. Then, the principles and recent developments of correlative SIM techniques are reviewed. Finally, the future development directions of SIM and its correlative microscopies are presented.

scholarly journals Spike Generators and Cell Signaling in the Human Auditory Nerve: An Ultrastructural, Super-Resolution, and Gene Hybridization Study

2021 ◽  
Vol 15 ◽  
Author(s):  
Wei Liu ◽  
Maria Luque ◽  
Hao Li ◽  
Anneliese Schrott-Fischer ◽  
Rudolf Glueckert ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Auditory Nerve ◽  
Spiral Ganglion ◽  
Super Resolution ◽  
Nerve Fibers ◽  
The Body ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Prosthetic Devices ◽  
Complex Sounds

Background: The human auditory nerve contains 30,000 nerve fibers (NFs) that relay complex speech information to the brain with spectacular acuity. How speech is coded and influenced by various conditions is not known. It is also uncertain whether human nerve signaling involves exclusive proteins and gene manifestations compared with that of other species. Such information is difficult to determine due to the vulnerable, “esoteric,” and encapsulated human ear surrounded by the hardest bone in the body. We collected human inner ear material for nanoscale visualization combining transmission electron microscopy (TEM), super-resolution structured illumination microscopy (SR-SIM), and RNA-scope analysis for the first time. Our aim was to gain information about the molecular instruments in human auditory nerve processing and deviations, and ways to perform electric modeling of prosthetic devices.Material and Methods: Human tissue was collected during trans-cochlear procedures to remove petro-clival meningioma after ethical permission. Cochlear neurons were processed for electron microscopy, confocal microscopy (CM), SR-SIM, and high-sensitive in situ hybridization for labeling single mRNA transcripts to detect ion channel and transporter proteins associated with nerve signal initiation and conductance.Results: Transport proteins and RNA transcripts were localized at the subcellular level. Hemi-nodal proteins were identified beneath the inner hair cells (IHCs). Voltage-gated ion channels (VGICs) were expressed in the spiral ganglion (SG) and axonal initial segments (AISs). Nodes of Ranvier (NR) expressed Nav1.6 proteins, and encoding genes critical for inter-cellular coupling were disclosed.Discussion: Our results suggest that initial spike generators are located beneath the IHCs in humans. The first NRs appear at different places. Additional spike generators and transcellular communication may boost, sharpen, and synchronize afferent signals by cell clusters at different frequency bands. These instruments may be essential for the filtering of complex sounds and may be challenged by various pathological conditions.

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 Metamaterial assisted illumination nanoscopy via random super-resolution speckles

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yeon Ui Lee ◽  
Junxiang Zhao ◽  
Qian Ma ◽  
Larousse Khosravi Khorashad ◽  
Clara Posner ◽  
...  
Keyword(s):  
Surface Characterization ◽  
Low Cost ◽  
Near Field ◽  
Super Resolution ◽  
Diffraction Limit ◽  
High Temporal Resolution ◽  
Wide Field ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Bio Imaging

AbstractStructured illumination microscopy (SIM) is one of the most powerful and versatile optical super-resolution techniques. Compared with other super-resolution methods, SIM has shown its unique advantages in wide-field imaging with high temporal resolution and low photon damage. However, traditional SIM only has about 2 times spatial resolution improvement compared to the diffraction limit. In this work, we propose and experimentally demonstrate an easily-implemented, low-cost method to extend the resolution of SIM, named speckle metamaterial-assisted illumination nanoscopy (speckle-MAIN). A metamaterial structure is introduced to generate speckle-like sub-diffraction-limit illumination patterns in the near field with improved spatial frequency. Such patterns, similar to traditional SIM, are then used to excite objects on top of the surface. We demonstrate that speckle-MAIN can bring the resolution down to 40 nm and beyond. Speckle-MAIN represents a new route for super-resolution, which may lead to important applications in bio-imaging and surface characterization.

scholarly journals Two-dimensional TIRF-SIM–traction force microscopy (2D TIRF-SIM-TFM)

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Liliana Barbieri ◽  
Huw Colin-York ◽  
Kseniya Korobchevskaya ◽  
Di Li ◽  
Deanna L. Wolfson ◽  
...  
Keyword(s):  
Resolution Enhancement ◽  
Traction Force ◽  
Super Resolution ◽  
Traction Force Microscopy ◽  
Two Dimensional ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Force Microscopy ◽  
Health And Disease ◽  
Spatio Temporal

AbstractQuantifying small, rapidly evolving forces generated by cells is a major challenge for the understanding of biomechanics and mechanobiology in health and disease. Traction force microscopy remains one of the most broadly applied force probing technologies but typically restricts itself to slow events over seconds and micron-scale displacements. Here, we improve >2-fold spatially and >10-fold temporally the resolution of planar cellular force probing compared to its related conventional modalities by combining fast two-dimensional total internal reflection fluorescence super-resolution structured illumination microscopy and traction force microscopy. This live-cell 2D TIRF-SIM-TFM methodology offers a combination of spatio-temporal resolution enhancement relevant to forces on the nano- and sub-second scales, opening up new aspects of mechanobiology to analysis.

scholarly journals Double moiré localized plasmon structured illumination microscopy

Nanophotonics ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Ruslan Röhrich ◽  
A. Femius Koenderink
Keyword(s):  
Near Field ◽  
Super Resolution ◽  
Reconstruction Algorithm ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Experimental Realization ◽  
Spatial Frequencies ◽  
Wide Range ◽  
Localized Plasmon ◽  
Plasmonic Grating

AbstractStructured illumination microscopy (SIM) is a well-established fluorescence imaging technique, which can increase spatial resolution by up to a factor of two. This article reports on a new way to extend the capabilities of structured illumination microscopy, by combining ideas from the fields of illumination engineering and nanophotonics. In this technique, plasmonic arrays of hexagonal symmetry are illuminated by two obliquely incident beams originating from a single laser. The resulting interference between the light grating and plasmonic grating creates a wide range of spatial frequencies above the microscope passband, while still preserving the spatial frequencies of regular SIM. To systematically investigate this technique and to contrast it with regular SIM and localized plasmon SIM, we implement a rigorous simulation procedure, which simulates the near-field illumination of the plasmonic grating and uses it in the subsequent forward imaging model. The inverse problem, of obtaining a super-resolution (SR) image from multiple low-resolution images, is solved using a numerical reconstruction algorithm while the obtained resolution is quantitatively assessed. The results point at the possibility of resolution enhancements beyond regular SIM, which rapidly vanishes with the height above the grating. In an initial experimental realization, the existence of the expected spatial frequencies is shown and the performance of compatible reconstruction approaches is compared. Finally, we discuss the obstacles of experimental implementations that would need to be overcome for artifact-free SR imaging.

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