Featured
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Research Highlight |
iSCAT gets a signal boost
Photonic resonator interferometric scattering microscopy offers label-free imaging of biomolecules in vitro with improved signal-to-noise ratio.
- Rita Strack
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Review Article |
Understanding the invisible hands of sample preparation for cryo-EM
The quality of structural data obtained in cryo-EM is affected by multiple factors pertaining to sample preparation. This Review discusses available techniques and current challenges.
- Giulia Weissenberger
- , Rene J. M. Henderikx
- & Peter J. Peters
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Article |
Deep learning-enhanced light-field imaging with continuous validation
A deep learning–based algorithm enables efficient reconstruction of light-field microscopy data at video rate. In addition, concurrently acquired light-sheet microscopy data provide ground truth data for training, validation and refinement of the algorithm.
- Nils Wagner
- , Fynn Beuttenmueller
- & Anna Kreshuk
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Article |
Phasor S-FLIM: a new paradigm for fast and robust spectral fluorescence lifetime imaging
Phasor S-FLIM combines novel electronics for multichannel fluorescence lifetime acquisition and a phasor-based unmixing algorithm for real-time analysis of reliable spectral lifetime imaging data, enabling new biological observations.
- Lorenzo Scipioni
- , Alessandro Rossetta
- & Enrico Gratton
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Research Highlight |
Expanding views of the transcriptome
In situ long-read sequencing combined with expansion microscopy enables precise views of transcriptomes in intact biological systems.
- Rita Strack
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Brief Communication |
Molecular-scale axial localization by repetitive optical selective exposure
ROSE-Z achieves axial interference through an asymmetrical optical scheme, yielding 2 nm axial localization precision with ~3,000 photons and a single objective, which offers improved multicolor three-dimensional localization microscopy for cellular structures.
- Lusheng Gu
- , Yuanyuan Li
- & Wei Ji
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Article |
Deep learning-based point-scanning super-resolution imaging
Point-scanning super-resolution imaging uses deep learning to supersample undersampled images and enable time-lapse imaging of subcellular events. An accompanying ‘crappifier’ rapidly generates quality training data for robust performance.
- Linjing Fang
- , Fred Monroe
- & Uri Manor
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Research Highlight |
Super-resolution 3D live cell imaging
3D pRESOLFT enables volumetric live cell imaging with high spatial and temporal resolution.
- Nina Vogt
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Correspondence |
Pycro-Manager: open-source software for customized and reproducible microscope control
- Henry Pinkard
- , Nico Stuurman
- & Laura Waller
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Article |
High-definition imaging using line-illumination modulation microscopy
HD-fMOST is a microscopy technique for imaging large samples at high throughput and with high definition, which is achieved with a line-illumination modulation approach. The technology is illustrated by imaging fluorescently labeled neurons in whole mouse brains.
- Qiuyuan Zhong
- , Anan Li
- & Qingming Luo
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Article |
Real-time volumetric reconstruction of biological dynamics with light-field microscopy and deep learning
Reconstruction of light-field microscopy data with a deep-learning network achieves high reconstruction speed and reduces artifacts, as illustrated for moving C. elegans and beating zebrafish hearts.
- Zhaoqiang Wang
- , Lanxin Zhu
- & Peng Fei
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Research Highlight |
DNA-PAINT takes a left turn
DNA-PAINT with left-handed DNA oligomers enables imaging of nuclear structures with reduced background.
- Rita Strack
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News & Views |
The promise and peril of deep learning in microscopy
- David P. Hoffman
- , Isaac Slavitt
- & Casey A. Fitzpatrick
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Method to Watch |
Single-objective light sheet microscopy
Single-objective light sheet fluorescence microscopes are driving innovation in volumetric imaging.
- Rita Strack
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Brief Communication |
Miniature two-photon microscopy for enlarged field-of-view, multi-plane and long-term brain imaging
A two-photon miniature microscope with enlarged field of view and axial scanning capabilities has been developed and applied in freely moving mice.
- Weijian Zong
- , Runlong Wu
- & Heping Cheng
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Article |
Click-ExM enables expansion microscopy for all biomolecules
Click-ExM uses click-chemistry-based labeling to increase the versatility of expansion microscopy. Click-ExM enables imaging of numerous classes of biomolecules including lipids, glycans, proteins, DNA, RNA and small molecules.
- De-en Sun
- , Xinqi Fan
- & Xing Chen
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Research Highlight |
Cryo-EM goes atomic
Technological advances push the limits of single-particle cryo-EM to achieve atomic resolution.
- Rita Strack
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Article |
Non-uniform refinement: adaptive regularization improves single-particle cryo-EM reconstruction
Membrane proteins exhibit spatial variation in rigidity and disorder, which poses a challenge for traditional cryo-EM reconstruction algorithms. Non-uniform refinement accounts for this spatial variability, yielding improved 3D reconstruction quality even for small membrane proteins.
- Ali Punjani
- , Haowei Zhang
- & David J. Fleet
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Editorial |
The power of a picture
This month, we celebrate the beauty and power of microscopy images.
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Article |
Live-cell super-resolved PAINT imaging of piconewton cellular traction forces
Tension-PAINT integrates molecular tension probes with DNA-PAINT to enable ~25-nm-resolution mapping of piconewton mechanical events. Tension-PAINT can be used to study dynamic forces, and an irreversible variant integrates force history over time.
- Joshua M. Brockman
- , Hanquan Su
- & Khalid Salaita
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This Month |
Manu Prakash
Frugally built technology to study the ocean’s microbes, and engineering for societal good.
- Vivien Marx
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Research Highlight |
A dual-constriction biological nanopore
Addition of a second constriction to CsgG-family nanopores improves the accuracy of homopolymer sequencing.
- Arunima Singh
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Correspondence |
SMAP: a modular super-resolution microscopy analysis platform for SMLM data
- Jonas Ries
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This Month |
Uri Manor
A new way to see actin in action, hearing as others don’t, and saved by the guitar.
- Vivien Marx
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Brief Communication |
Optimizing imaging speed and excitation intensity for single-molecule localization microscopy
A systematic evaluation shows that excitation intensity has a dramatic impact on image quality in localization microscopy and reveals the benefits of lower excitation intensity for improved labeling efficiency and localization precision.
- Robin Diekmann
- , Maurice Kahnwald
- & Jonas Ries
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Article |
Scale-free vertical tracking microscopy
Scale-free vertical tracking microscopy based on a ‘hydrodynamic treadmill’ enables measuring long-range movements of freely suspended organisms with high spatiotemporal resolution.
- Deepak Krishnamurthy
- , Hongquan Li
- & Manu Prakash
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Brief Communication |
Time-resolved cryo-EM using Spotiton
A spraying-mixing approach for preparing cryo-EM grids using the Spotiton robot allows time-resolved observations of short-lived biomolecular states.
- Venkata P. Dandey
- , William C. Budell
- & Bridget Carragher
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Brief Communication |
Actin chromobody imaging reveals sub-organellar actin dynamics
Genetically encoded fluorescent-protein-tagged actin nanobodies fused to organelle membrane-targeting sequences enables high-resolution imaging of transient organelle–actin contact sites in live cells.
- Cara R. Schiavon
- , Tong Zhang
- & Uri Manor
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Research Highlight |
Imaging single glycan molecules
Direct imaging of carbohydrates using electrospray ion beam deposition and scanning tunneling microscopy.
- Arunima Singh
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Article |
ZipSeq: barcoding for real-time mapping of single cell transcriptomes
ZipSeq uses patterned illumination and photocaged oligonucleotides to serially print barcodes onto live, intact tissues for spatially mapping single cell transcriptomes.
- Kenneth H. Hu
- , John P. Eichorst
- & Matthew F. Krummel
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News & Views |
A new way to see RNAs
Producing reliable atomic- or close-to-atomic-resolution structures of RNA-only molecules has been a formidable task. Ribosolve can solve sub-nanometer-resolution cryo-EM structures of unbound RNA molecules with unprecedented accuracy and speed.
- Jane S. Richardson
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Brief Communication |
Up to 100-fold speed-up and multiplexing in optimized DNA-PAINT
Hundred-fold-faster DNA-PAINT imaging is enabled by the introduction of concatenated, periodic DNA sequence motifs in the docking strand. Six orthogonal sequences are described for speed-optimized and highly multiplexed cellular imaging.
- Sebastian Strauss
- & Ralf Jungmann
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Article |
Homogeneous multifocal excitation for high-throughput super-resolution imaging
Multifocal flat illumination for field-independent imaging (mfFIFI) enables patterned illumination over an extended field of view. Integration with instant structured illumination microscope allowed for high-speed, multicolor, volumetric super-resolution imaging over 100 × 100 µm2.
- Dora Mahecic
- , Davide Gambarotto
- & Suliana Manley
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Article |
DeepSTORM3D: dense 3D localization microscopy and PSF design by deep learning
DeepSTORM3D uses deep learning for accurate localization of point emitters in densely labeled samples in three dimensions for volumetric localization microscopy with high temporal resolution, as well as for optimal point-spread function design.
- Elias Nehme
- , Daniel Freedman
- & Yoav Shechtman
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Article |
Real-time 3D movement correction for two-photon imaging in behaving animals
Real-time 3D movement correction by tracking a fluorescent bead in the field of view enables functional imaging with 3D two-photon random-access microscopy in behaving mice and zebrafish.
- Victoria A. Griffiths
- , Antoine M. Valera
- & R. Angus Silver
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Brief Communication |
Three-photon head-mounted microscope for imaging deep cortical layers in freely moving rats
A head-mounted three-photon microscope based on a custom-designed optical fiber and dispersion compensation enables imaging of activity from neuronal populations deep in the cortex of freely moving rats.
- Alexandr Klioutchnikov
- , Damian J. Wallace
- & Jason N. D. Kerr
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Article |
Three-dimensional nanoscopy of whole cells and tissues with in situ point spread function retrieval
In situ point spread function (PSF) retrieval (INSPR) enables precise single-molecule localization in 3D single-molecule localization microscopy of whole cells and tissues. It directly determines PSF from a single-molecule blinking dataset, removing errors associated with sample-induced aberrations.
- Fan Xu
- , Donghan Ma
- & Fang Huang
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Brief Communication |
Deep three-photon imaging of the brain in intact adult zebrafish
Three-photon microscopy provides access to most of the adult zebrafish brain for both structural and functional imaging, as well as to the Danionella dracula brain.
- Dawnis M. Chow
- , David Sinefeld
- & Joseph R. Fetcho
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Review Article |
Scanless two-photon excitation with temporal focusing
This Review discusses temporal focusing microscopy and its applications in neuroscience for imaging and optogenetic activation.
- Eirini Papagiakoumou
- , Emiliano Ronzitti
- & Valentina Emiliani
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Article |
Single-molecule displacement mapping unveils nanoscale heterogeneities in intracellular diffusivity
Single-molecule displacement/diffusivity mapping (SMdM) enables nanoscale mapping of freely diffusing molecules in mammalian cells and reveals the structural basis of variations in local diffusivity in both the cytoplasm and nucleus.
- Limin Xiang
- , Kun Chen
- & Ke Xu
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PYMEVisualize: an open-source tool for exploring 3D super-resolution dataBrowse broader subjects
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- Atomic force microscopy
- Ca2+ imaging
- Confocal microscopy
- Cryoelectron microscopy
- Interference microscopy
- Light-sheet microscopy
- Multiphoton microscopy
- Phase-contrast microscopy
- Polarization microscopy
- Scanning electron microscopy
- Scanning probe microscopy
- Super-resolution microscopy
- Total internal reflection microscopy
- Transmission electron microscopy
- Transmission light microscopy
- Wide-field fluorescence microscopy
