Microscopy

  • Article
    | Open Access

    Although the first dissection of the human ovary dates back to the 17th century, its characterization is still limited. Here, the authors have unraveled a unique biophysical and topological phenotype of reproductive-age tissue, bridging biophysics and female fertility and providing a blueprint for the artificial ovary.

    • Emna Ouni
    • , Alexis Peaucelle
    •  & Christiani A. Amorim
  • Article
    | Open Access

    Determining the quality of localisation microscopy images is currently challenging. Here the authors report use of the Haar wavelet kernel analysis (HAWK) Method for the Assessment of Nanoscopy, termed HAWKMAN, to assess the reliability of localisation information.

    • Richard J. Marsh
    • , Ishan Costello
    •  & Susan Cox
  • Article
    | Open Access

    The behaviour of Tungsten ditelluride (WTe2) in few-layer form is not yet fully characterized. Here the authors use a near-field terahertz microscopy technique to observe the electromagnetic responses of WTe2 flakes from one to several layers and to study their semimetallic/ semiconducting behavior.

    • Ran Jing
    • , Yinming Shao
    •  & D. N. Basov
  • Article
    | Open Access

    Molecular tethers physically bridge transport vesicles to their target membranes as a prerequisite step for fusion. Here the authors control vesicle tethering using optogenetic approaches to study the interplay between vesicle tethering and fusion.

    • Seong J. An
    • , Felix Rivera-Molina
    •  & Derek Toomre
  • Article
    | Open Access

    The morphology of semicrystalline plastics on the 1-100 μm scale, such as spherulites, strongly affect mechanical and other properties of the material but currently only 2D imaging techniques are available. Here, the authors use fluorescence labels and confocal microscopy to visualize the internal structure of neat polymers and composites in 3D and reveal unsuspected morphologies.

    • Shu-Gui Yang
    • , Zhen-Zhen Wei
    •  & Goran Ungar
  • Article
    | Open Access

    The authors introduce an analytical approach for quantitative analysis of 3D atom dynamics during electron microscopy. They image a Co-Mo-S nanocrystal with 1.5 Å resolution, and observe chemical transformations caused by beam-stimulated vibrations.

    • Fu-Rong Chen
    • , Dirk Van Dyck
    •  & Stig Helveg
  • Article
    | Open Access

    Single-pixel holography generates holographic images with a single-pixel detector making this relatively inexpensive. Here the authors report a high-throughput single-pixel compressive holography method for imaging biological tissue which can either provide a large field of view or high resolution.

    • Daixuan Wu
    • , Jiawei Luo
    •  & Zhaohui Li
  • Article
    | Open Access

    The authors have recently developed molecular force microscopy (MFM) which uses fluorescence polarisation to measure cell-surface receptor force orientation. Here they show that structured illumination microscopes, which inherently use fluorescence polarisation, can be used for MFM in a turn-key manner.

    • Aaron Blanchard
    • , J. Dale Combs
    •  & Khalid Salaita
  • Article
    | Open Access

    Honeycomb layered oxides are an emerging class of materials with peculiar physicochemical properties. Here, the authors report the synthesis and electrochemical energy storage characterisations of a mixed-alkali honeycomb layered oxide material capable of storing Na and K ions simultaneously.

    • Titus Masese
    • , Yoshinobu Miyazaki
    •  & Tomohiro Saito
  • Article
    | Open Access

    The authors demonstrate efficient excitation of nanodiamonds by a focused beam of helium ions, resulting in ionoluminescence. They use this for quantification and correlative localization of single particles within a whole cell at sub-30 nm resolution, and investigate nanodiamond radiosensitisation effects.

    • Zhaohong Mi
    • , Ce-Belle Chen
    •  & Andrew A. Bettiol
  • Article
    | Open Access

    Mitochondrial transport toward both the plus- and minus-ends of microtubules is mediated by motor proteins linked to mitochondria by TRAK adaptor proteins. Here the authors investigate the role of TRAK2 as a bidirectional motor adaptor, and propose a model where TRAK2 coordinates the activities of opposing kinesin-1 and cytoplasmic dynein motors as a single interdependent motor complex.

    • Adam R. Fenton
    • , Thomas A. Jongens
    •  & Erika L. F. Holzbaur
  • Article
    | Open Access

    Imaging rates in single-pixel imaging has been limited by the dependence on configurable spatial light modulators. Here, the authors use cyclic Hadamard patterns coded onto a spinning mask to demonstrate dynamic imaging with rates up to 72 frames per second and real time reconstruction capabilities.

    • Evgeny Hahamovich
    • , Sagi Monin
    •  & Amir Rosenthal
  • Article
    | Open Access

    The authors develop a method to build Manhattan Raman Scattering (MARS) probes based on different core atoms, conjugation ring numbers, and stable isotope substitutions. A quantitative model predicts vibrational frequencies of MARS dyes from structures, which are used in supermultiplexed vibrational imaging.

    • Yupeng Miao
    • , Naixin Qian
    •  & Wei Min
  • Article
    | Open Access

    Currently, bidirectional control of activity in the same neurons in the same experiment is difficult. Here the authors report a Bidirectional Pair of Opsins for Light-induced Excitation and Silencing, BiPOLES, which they use in a range of organisms including worms, fruit flies, mice and ferrets.

    • Johannes Vierock
    • , Silvia Rodriguez-Rozada
    •  & J. Simon Wiegert
  • Article
    | Open Access

    Wavefront shaping is used to overcome scattering in biological tissues during imaging, but determining the compensation is slow. Here, the authors use holographic phase stepping interferometry, where new phase information is updated after each measurement, enabling fast improvement of the wavefront correction.

    • Molly A. May
    • , Nicolas Barré
    •  & Alexander Jesacher
  • Article
    | Open Access

    Pleckstrin homology (PH) domains are presumed to bind phosphoinositides (PIPs) but only few specific interactions are known. Using TIRF microscopy based assays, functional studies and an experimentally validated prediction algorithm, the authors show that specific PIP binding is widespread among human PH domains.

    • Nilmani Singh
    • , Adriana Reyes-Ordoñez
    •  & Jie Chen
  • Article
    | Open Access

    3D real-space analysis of thick nanoparticle crystals is non-trivial. Here, the authors demonstrate the structural analysis of a bulk-like Laves phase by imaging an off-stoichiometric binary mixture of hard-sphere-like nanoparticles in spherical confinement by electron tomography, enabling defect analysis on the single-particle level.

    • Da Wang
    • , Ernest B. van der Wee
    •  & Alfons van Blaaderen
  • Article
    | Open Access

    The authors demonstrate accurate localization in three dimensions by comprehensive calibration of an ordinary microscope, exploiting the latent information of intrinsic aberrations. Rigid transformation of the emitter positions tests the method and enables measurements in six degrees of freedom.

    • Craig R. Copeland
    • , Craig D. McGray
    •  & Samuel M. Stavis
  • Article
    | Open Access

    Particle fusion can improve signal-to-noise ratio in single molecule localization microscopy, but is limited by structural heterogeneity. Here, the authors demonstrate an unsupervised classification method that differentiates structurally different DNA origami structures without prior knowledge.

    • Teun A.P.M. Huijben
    • , Hamidreza Heydarian
    •  & Bernd Rieger
  • Article
    | Open Access

    Super-resolution microscopy is often limited by photobleaching or uneven distribution of fluorophores. The authors present a label-free superresolution method termed VISTA, combining sample-expansion and vibrational imaging, with resolution down to 78 nm in protein-rich biological structures in cells and tissues.

    • Chenxi Qian
    • , Kun Miao
    •  & Lu Wei
  • Article
    | Open Access

    Studying dynamic processes in mechanobiology has been challenging due to lack of appropriate tools. Here, the authors present an interference-based method, illuminated via two rapidly alternating wavelengths, which enables real-time mapping of nanoscale forces with sub-second mechanical fluctuations.

    • Andrew T. Meek
    • , Nils M. Kronenberg
    •  & Malte C. Gather
  • Article
    | Open Access

    Cyanobacterial thylakoid membranes host the molecular machinery for the light-dependent reactions of photosynthesis and respiratory electron flow. Here, the authors show that newly synthesized thylakoids emerge between the plasma membrane and pre-existing thylakoids and describe the time-dependent assembly process of photosynthetic complexes.

    • Tuomas Huokko
    • , Tao Ni
    •  & Lu-Ning Liu
  • Article
    | Open Access

    It is difficult to apply SMLM to complex biological tissues. Here the authors report REALM, Robust and Effective Adaptive Optics in Localisation Microscopy, to improve SMLM in tissue and use this to resolve the organisation of spectrin in the axon initial segment in brain tissue.

    • Marijn E. Siemons
    • , Naomi A. K. Hanemaaijer
    •  & Lukas C. Kapitein
  • Article
    | Open Access

    Probes with reversible fluorescence are useful in super-resolution microscopy, but lack sufficient chemical specificity. Here, the authors engineer alkyne tagged diarylethene to realize photo-switchable stimulated Raman scattering probes with high chemical resolution, for applications in living cells.

    • Jianpeng Ao
    • , Xiaofeng Fang
    •  & Minbiao Ji
  • Article
    | Open Access

    The authors introduce stochastic optical scattering localization imaging (SOSLI) for non-invasive super-resolution imaging through scattering media. They capture multiple speckle patterns of photo-switchable point sources and use the speckle correlation to retrieve images with 100 nm resolution.

    • Dong Wang
    • , Sujit K. Sahoo
    •  & Cuong Dang
  • Article
    | Open Access

    Optical aberrations in Structured Illumination Microscopy (SIM) can lead to loss of resolution and artifacts making it unsuitable for thick samples. Here the authors combine Adaptive Optics and SIM (AO-3DSIM) to improve the 3D resolution and reduce artifacts, performing 3D-SIM in C.elegans.

    • Ruizhe Lin
    • , Edward T. Kipreos
    •  & Peter Kner
  • Article
    | Open Access

    Uniform illumination is a prerequisite for quantitative analyses in both classical fluorescence microscopy and single molecule localisation microscopy. Here, the authors introduce ASTER, an illumination technique that generates uniform illumination over large and adaptable fields of view, compatible with epifluorescence, HiLo and TIRF illumination schemes.

    • Adrien Mau
    • , Karoline Friedl
    •  & Sandrine Lévêque-Fort
  • Article
    | Open Access

    Diffractive optical elements are broadly used due to their ability to reshape the wavefront efficiently and conveniently. By using liquid immersion, the authors here enable microscale 3D-printed optics to behave like nanoscopic structures while maintaining high performance.

    • Reut Orange-Kedem
    • , Elias Nehme
    •  & Yoav Shechtman
  • Article
    | Open Access

    Here, the authors present a high-speed photothermal spatial light modulator which can generate a step-like wavefront change without diffraction artifacts. They use this to perform quantitative phase imaging, capturing sub-millisecond motion with a nanometer resolution in 3D.

    • Hadrien M. L. Robert
    • , Kristýna Holanová
    •  & Marek Piliarik
  • Article
    | Open Access

    Adaptation of current algorithms to 3D SMLM data is currently problematic. Here the authors report a method that increases the signal-to-noise ratio and resolution of 3D single particle analysis in localization microscopy and enables determination of the symmetry groups of macromolecular complexes.

    • Hamidreza Heydarian
    • , Maarten Joosten
    •  & Bernd Rieger
  • Article
    | Open Access

    The FRET efficiency usually predominantly depends on the proximity of donor and acceptor. Here the authors report an anisotropy-based mode of FRET detection, FRET-induced Angular Displacement Evaluation via Dim donor (FADED), to allow quantification of the relative angle between donor and acceptor.

    • Danai Laskaratou
    • , Guillermo Solís Fernández
    •  & Hideaki Mizuno
  • Article
    | Open Access

    Previous work has demonstrated electric-field detection with nitrogen-vacancy centers in diamond; however, nanoscale electric-field imaging has not been shown. Here, the authors use individual nitrogen-vacancy centers to map out electric field contours from a tip of an atomic force microscope with 10 nm resolution.

    • Ke Bian
    • , Wentian Zheng
    •  & Ying Jiang
  • Article
    | Open Access

    Bacterial cell division by cell wall synthesis proteins is guided by treadmilling filaments of the cytoskeleton protein FtsZ. Here authors use nanofabrication, advanced microscopy, and microfluidics to resolve the function of FtsZ treadmilling in the Gram-positive model organism Bacillus subtilis.

    • Kevin D. Whitley
    • , Calum Jukes
    •  & Séamus Holden
  • Article
    | Open Access

    Encoding data in DNA is a promising approach to high density data storage. Here the authors present a prototype sequencing-free method that uses the spatial orientation of DNA strands with super-resolution microscopy readout.

    • George D. Dickinson
    • , Golam Md Mortuza
    •  & William L. Hughes
  • Article
    | Open Access

    Space charges or injected charges dominate the ferroelectric degradation, however, its physics insight remains unclear. Here, the authors reveal the nature of ferroelectric degradation in nanodevices and the role of the injected charges in polarization reversal.

    • Qianwei Huang
    • , Zibin Chen
    •  & Xiaozhou Liao
  • Article
    | Open Access

    Performing multiple FRET measurements at once can be challenging. Here the authors report a method to discriminate between overlapping FRET pairs, even if the fluorophores display almost identical absorption and emission spectra, based on the photochromism of the donor fluorophores.

    • Thijs Roebroek
    • , Wim Vandenberg
    •  & Peter Dedecker
  • Article
    | Open Access

    Multiphoton microscopy requires precise increases in excitation power with imaging depth to generate contrast without damaging the sample. Here the authors show how an adaptive illumination function can be learned from the sample’s shape and used for in vivo imaging of whole lymph nodes.

    • Henry Pinkard
    • , Hratch Baghdassarian
    •  & Laura Waller
  • Article
    | Open Access

    Super-resolution microscopy and single molecule fluorescence spectroscopy require optimisation of the temporal or spatial resolution, which are usually mutually exclusive. Here the authors report a GPU-supported, camera-based strategy to achieve high spatial and temporal resolution from the same dataset.

    • Jagadish Sankaran
    • , Harikrushnan Balasubramanian
    •  & Thorsten Wohland
  • Article
    | Open Access

    Here, the authors present photonic resonator interferometric scattering microscopy, which utilises a dielectric photonic crystal as the sample substrate. The resonant near-field enhancement leads to improved signal to noise ratio without increasing illumination intensity.

    • Nantao Li
    • , Taylor D. Canady
    •  & Brian T. Cunningham
  • Article
    | Open Access

    Structured illumination microscopy is usually limited to 2 times spatial resolution improvement over the diffraction limit. Here, the authors introduce a metamaterial structure to generate speckle-like sub-diffraction limit illumination patterns in the near field, and achieve a 7-fold resolution improvement down to 40 nm.

    • Yeon Ui Lee
    • , Junxiang Zhao
    •  & Zhaowei Liu
  • Article
    | Open Access

    Here, the authors present two local methods for analyzing cryo-EM maps: LocSpiral and LocBSharpen that enhance high-resolution features of cryoEM maps, while preventing map distortions. They also introduce LocBFactor and LocOccupancy, which allow obtaining local B-factors and electron density occupancy maps from cryo-EM reconstructions and the authors demonstrate that these methods improve the interpretability and analysis of cryo-EM maps using different test cases among them recent SARS-CoV-2 spike glycoprotein structures.

    • Satinder Kaur
    • , Josue Gomez-Blanco
    •  & Javier Vargas