Featured
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Article |
Large-scale deep tissue voltage imaging with targeted-illumination confocal microscopy
In targeted-illumination confocal microscopy, the combination of targeted illumination with confocal detection allows one-photon voltage imaging across large fields of view with a high signal-to-noise ratio.
- Sheng Xiao
- , William J. Cunningham
- & Jerome Mertz
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Research Briefing |
VIBRANT: a phenotyping method for drug discovery using vibrational spectroscopy
We developed a high-content profiling method named vibrational painting (VIBRANT) for single-cell drug response measurements, combining vibrational imaging, multiplexed vibrational probes and machine learning. VIBRANT showed high performance in predicting drug mechanisms of action, discovering novel compounds and assessing drug combinations, demonstrating great promise for phenotypic drug discovery.
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Article |
VIBRANT: spectral profiling for single-cell drug responses
Vibrational painting (VIBRANT) is a high-content single-cell phenotypic profiling method using mid-infrared imaging with vibrational probes for metabolic activity, which offers high accuracy with minimal batch effects to capture cellular responses to perturbation.
- Xinwen Liu
- , Lixue Shi
- & Wei Min
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Article |
Crystal ribcage: a platform for probing real-time lung function at cellular resolution
A fabricated transparent ribcage facilitates real-time analysis of whole-lung function down to cellular resolution.
- Rohin Banerji
- , Gabrielle N. Grifno
- & Hadi T. Nia
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Brief Communication
| Open AccessOrgan Mapping Antibody Panels: a community resource for standardized multiplexed tissue imaging
Organ Mapping Antibody Panels are a community-led initiative to create standardized antibody panels for multiplexed spatial imaging.
- Ellen M. Quardokus
- , Diane C. Saunders
- & Andrea J. Radtke
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Article
| Open AccessCross-modality supervised image restoration enables nanoscale tracking of synaptic plasticity in living mice
XTC is a supervised deep-learning-based image-restoration approach that is trained with images from different modalities and applied to an in vivo modality with no ground truth. XTC’s capabilities are demonstrated in synapse tracking in the mouse brain.
- Yu Kang T. Xu
- , Austin R. Graves
- & Jeremias Sulam
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News & Views |
Line-scanning speeds up Brillouin microscopy
Two new Brillouin microscopes leverage line-scanning to overcome previous limitations of the technique, enabling fast imaging, with low phototoxicity, of mechanical properties in living embryos of model organisms and tumor spheroids.
- Nargess Khalilgharibi
- , Giulia Paci
- & Yanlan Mao
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Article
| Open AccessHigh-resolution line-scan Brillouin microscopy for live imaging of mechanical properties during embryo development
Line-scan Brillouin microscopy enables fast 3D imaging of mechanical properties with low phototoxicity, as shown for Drosophila and mouse embryos, as well as ascidians.
- Carlo Bevilacqua
- , Juan Manuel Gomez
- & Robert Prevedel
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Brief Communication |
LILAC: enhanced actin imaging with an optogenetic Lifeact
LILAC is a photoactivatable version of Lifeact, a tool for labeling F-actin. LILAC can help avoid cytotoxicity, which is sometimes associated with the use of Lifeact.
- Kourtney L. Kroll
- , Alexander R. French
- & Ronald S. Rock
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Article
| Open AccessHyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal-to-noise fluorescence
Hybrid Unmixing offers enhanced imaging of multiplexed fluorescence labels, enabling longitudinal imaging of multiple fluorescent signals with reduced illumination intensities.
- Hsiao Ju Chiang
- , Daniel E. S. Koo
- & Francesco Cutrale
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Article
| Open AccessOptimizing multiplexed imaging experimental design through tissue spatial segregation estimation
A statistical approach for optimal design of multiplexed imaging studies has been developed. It determines experimental parameters that facilitate cell phenotype identification.
- Pierre Bost
- , Daniel Schulz
- & Bernd Bodenmiller
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Brief Communication |
Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein
miRFP718nano is a rationally designed small near-infrared fluorescent protein with an emission tail that extends into the short-wave infrared range for improved multiplexed and deep-tissue imaging applications.
- Olena S. Oliinyk
- , Chenshuo Ma
- & Vladislav V. Verkhusha
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Article
| Open AccessImage-seq: spatially resolved single-cell sequencing guided by in situ and in vivo imaging
Image-seq isolates cells from specific tissue locations under image guidance for analysis by single-cell RNA sequencing. The technique can be combined with in vivo imaging to document the temporal and dynamic history of the cells prior to sequencing.
- Christa Haase
- , Karin Gustafsson
- & Charles P. Lin
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Article |
Multiplexed bioluminescence microscopy via phasor analysis
The combination of engineered probes and spectral phasor analysis overcomes long-standing challenges associated with bioluminescence detection at the microscale, enabling multiplexed, real-time imaging of cellular features without the need for excitation light.
- Zi Yao
- , Caroline K. Brennan
- & Jennifer A. Prescher
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Article |
Automated high-speed 3D imaging of organoid cultures with multi-scale phenotypic quantification
A method for high-content 3D imaging of organoids.
- Anne Beghin
- , Gianluca Grenci
- & Virgile Viasnoff
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Method to Watch |
Imaging without the labels
Advances in label-free microscopy are expanding experimental observations, from biophysics to cell biology and beyond.
- Rita Strack
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Editorial |
Minding microscopy metadata
Guidelines for improving reporting and reproduciblity in microscopy take center stage in this month’s Focus issue.
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Article |
An adaptive optics module for deep tissue multiphoton imaging in vivo
A compact adaptive optics module corrects aberrations in two-photon and three-photon microscopy, enabling structural and functional imaging deep in the mouse brain, the mouse spinal cord and the zebrafish larva.
- Cristina Rodríguez
- , Anderson Chen
- & Na Ji
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Article |
High-speed, cortex-wide volumetric recording of neuroactivity at cellular resolution using light beads microscopy
Light beads microscopy is a two-photon microscopy approach that allows high-speed volumetric imaging of neuronal activity at the mesoscale.
- Jeffrey Demas
- , Jason Manley
- & Alipasha Vaziri
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Perspective |
Best practices and tools for reporting reproducible fluorescence microscopy methods
Comprehensive guidelines and resources to enable accurate reporting for the most common fluorescence light microscopy modalities are reported with the goal of improving microscopy reporting, rigor and reproducibility.
- Paula Montero Llopis
- , Rebecca A. Senft
- & Michelle S. Itano
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Article |
Three-dimensional adaptive optical nanoscopy for thick specimen imaging at sub-50-nm resolution
The combination of adaptive optics with an improved isoSTED nanoscope allows imaging of cells and tissues with sub-50-nm isotropic resolution.
- Xiang Hao
- , Edward S. Allgeyer
- & Joerg Bewersdorf
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Comment |
REMBI: Recommended Metadata for Biological Images—enabling reuse of microscopy data in biology
Bioimaging data have significant potential for reuse, but unlocking this potential requires systematic archiving of data and metadata in public databases. We propose draft metadata guidelines to begin addressing the needs of diverse communities within light and electron microscopy. We hope this publication and the proposed Recommended Metadata for Biological Images (REMBI) will stimulate discussions about their implementation and future extension.
- Ugis Sarkans
- , Wah Chiu
- & Alvis Brazma
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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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News & Views |
Deep-learning-enhanced light-field microscopy
Light-field microscopes can image three-dimensional dynamics of biological samples at unprecedented speed, but the computational reconstruction necessary for image formation is artifact-prone and time-consuming. Deep learning closes this gap between imaging and reconstruction speed.
- Kai Wang
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Article |
Miniaturized head-mounted microscope for whole-cortex mesoscale imaging in freely behaving mice
The mini-mScope is a miniature microscope that can image neural activity at the mesoscale in most of the dorsal cortex of freely behaving mice.
- Mathew L. Rynes
- , Daniel A. Surinach
- & Suhasa B. Kodandaramaiah
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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 |
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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Article |
Next-generation GRAB sensors for monitoring dopaminergic activity in vivo
Red and improved green versions of the genetically encoded dopamine sensor GRABDA have been developed. These neurotransmitter sensors are used alone or in combination with, for example, calcium sensors in behaving fruit flies and rodents.
- Fangmiao Sun
- , Jingheng Zhou
- & Yulong Li
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Article |
Plasmonic scattering imaging of single proteins and binding kinetics
Plasmonic scattering microscopy (PSM) enables the imaging of single proteins on SPR instruments. The method enables measurement of protein size and binding kinetics and is fully compatible with simultaneous traditional SPR measurements.
- Pengfei Zhang
- , Guangzhong Ma
- & Nongjian Tao
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Brief Communication |
High-sensitivity and high-specificity biomechanical imaging by stimulated Brillouin scattering microscopy
Stimulated Brillouin scattering microscopy overcomes the trade-off between acquisition speed and spectral resolution in spontaneous Brillouin scattering microscopy and allows visualization of elasticity and viscosity, as shown in C. elegans.
- Itay Remer
- , Roni Shaashoua
- & Alberto Bilenca
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Article |
Mid-infrared metabolic imaging with vibrational probes
Small vibrational tags (azide, 13C-edited carbonyl and deuterium-labeled probes) were introduced as metabolic probes for mid-infrared imaging. The tags allow unprecedented in situ visualization of metabolism in cells and animals with high information throughput.
- Lixue Shi
- , Xinwen Liu
- & Wei Min
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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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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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Brief Communication |
An adaptive excitation source for high-speed multiphoton microscopy
An adaptive excitation source enables two- and three-photon imaging of the awake mouse brain with high spatial and temporal resolution at 30-fold-reduced laser power relative to conventional approaches.
- Bo Li
- , Chunyan Wu
- & Chris Xu
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Brief Communication |
Non-reversible tissue fixation retains extracellular vesicles for in situ imaging
Tissue fixation with formaldehyde and a water-soluble carbodiimide crosslinker (EDC) leads to retention of extracellular vesicles within tissues and allows for reliable extracellular vesicle imaging for semiquantitative imaging applications.
- Mrinali P. Gupta
- , Sangeetha Tandalam
- & John T. G. Pena
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Comment |
Putting advanced microscopy in the hands of biologists
The growth of advanced, custom microscopy has outpaced commercialization, with biologists unable to benefit from these developments. We propose a complementary model for access based on shareable, traveling and configurable microscopes, with potential benefits for biologists, developers and the community.
- Rory M. Power
- & Jan Huisken
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Brief Communication |
An order of magnitude faster DNA-PAINT imaging by optimized sequence design and buffer conditions
DNA-PAINT is sped up by an order of magnitude by optimizing sequences and buffer conditions, enabling faster imaging with no compromise to image quality or resolution, improved single-molecule counting and enhanced cellular imaging.
- Florian Schueder
- , Johannes Stein
- & Ralf Jungmann
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Article |
Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0
SCAPE 2.0 is a versatile imaging platform that enables real-time three-dimensional microscopy of cellular function and dynamic motion in living organisms at over 100 volumes per second with minimal photodamage, and high-throughput structural imaging in fixed, cleared and expanded samples.
- Venkatakaushik Voleti
- , Kripa B. Patel
- & Elizabeth M. C. Hillman
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Review Article |
Brillouin microscopy: an emerging tool for mechanobiology
This Review discusses the principles, advantages and limitations of Brillouin microscopy, a non-invasive tool for measuring mechanical properties of biological samples in three dimensions.
- Robert Prevedel
- , Alba Diz-Muñoz
- & Giuseppe Antonacci
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Brief Communication |
Molecular resolution imaging by repetitive optical selective exposure
Repetitive optical selective exposure (ROSE) is an interferometric single-molecule localization microscopy method offering twofold improvement in lateral resolution with the same photon budget compared with conventional approaches.
- Lusheng Gu
- , Yuanyuan Li
- & Wei Ji
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Perspective |
Biological imaging of chemical bonds by stimulated Raman scattering microscopy
This Perspective summarizes advances in stimulated Raman spectroscopy (SRS) instruments and probes, and highlights biological discoveries made with these technologies.
- Fanghao Hu
- , Lixue Shi
- & Wei Min
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Brief Communication |
Thermophoretic trap for single amyloid fibril and protein aggregation studies
A thermophoretic trapping approach enables users to observe the behavior of single amyloid fibrils in solution for hours without any immobilization.
- Martin Fränzl
- , Tobias Thalheim
- & Frank Cichos
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Brief Communication |
Direct wavefront sensing enables functional imaging of infragranular axons and spines
Two-photon microscopy in combination with adaptive optics enables diffraction-limited morphological and functional imaging up to around 800 μm below the pia. This is achieved with the help of fluorescent microvessels serving as guidestars.
- Rui Liu
- , Zengyi Li
- & David Kleinfeld
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Research Highlight |
Scattering microscopy takes single-particle tracking to the next level
Interferometric scattering microscopy enables extended monitoring of gold-nanoparticle-labeled membrane proteins with impressive spatiotemporal resolution.
- Rita Strack