News & Views |
Featured
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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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Research Highlight |
CLEM takes a polar plunge
Combining cryogenic super-resolution microscopy and focused ion beam scanning electron microscopy allows nanoscale views of protein architecture in the context of cellular ultrastructure in whole cells.
- Rita Strack
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This Month |
Na Ji
How joy and wide-ranging curiosity leads to neurobiology tools for new types of questions.
- Vivien Marx
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Brief Communication |
Rapid mesoscale volumetric imaging of neural activity with synaptic resolution
Integrating a Bessel focus module into a two-photon fluorescence mesoscope enables high-speed volumetric imaging of neuronal activity in soma, dendrites and spines.
- Rongwen Lu
- , Yajie Liang
- & Na Ji
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Brief Communication |
Kilohertz two-photon fluorescence microscopy imaging of neural activity in vivo
High-speed two-photon laser scanning microscopy using a passive laser scanner based on free-space angular-chirp-enhanced delay achieves frame rates suitable for voltage imaging in vivo in the mouse brain.
- Jianglai Wu
- , Yajie Liang
- & Na Ji
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Perspective |
Visualizing the genome in high resolution challenges our textbook understanding
This Perspective highlights how high-resolution imaging has informed our view and helped overturn the textbook understanding of 4D genome organization.
- Melike Lakadamyali
- & Maria Pia Cosma
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Brief Communication |
Simultaneous multiplane imaging with reverberation two-photon microscopy
Reverberation two-photon microscopy enables video-rate multiplane neuroimaging by performing near-instantaneous axial scanning over large depth ranges while maintaining 3D micrometer-scale resolution.
- Devin R. Beaulieu
- , Ian G. Davison
- & Jerome Mertz
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Article |
MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells
Advances in MINFLUX nanoscopy enable multicolor imaging over large fields of view, bringing true nanometer-scale fluorescence imaging to labeled structures in fixed and living cells.
- Klaus C. Gwosch
- , Jasmin K. Pape
- & Stefan W. Hell
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Research Highlight |
Be still, my beating heart
An optical gating strategy allows day-long light-sheet imaging of the developing heart in zebrafish embryos.
- Rita Strack
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Research Highlight |
RAMPing up voltage indicator imaging
Combining a voltage sensor with random-access microscopy allows imaging of neuronal activity in behaving mice.
- Nina Vogt
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Brief Communication |
Localization microscopy at doubled precision with patterned illumination
SIMFLUX combines elements of MINFLUX with structured illumination to double localization precision and improve resolution in localization microscopy. The approach was demonstrated on DNA origami and on cellular microtubules.
- Jelmer Cnossen
- , Taylor Hinsdale
- & Sjoerd Stallinga
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