Featured
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Methods in Brief |
Finding and correcting immobile fluorophores
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Correspondence |
Fluorescence and super-resolution standards based on DNA origami
- Jürgen J Schmied
- , Andreas Gietl
- & Philip Tinnefeld
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Brief Communication |
Ultrabright photoactivatable fluorophores created by reductive caging
The resolution achievable with single-molecule–based super-resolution fluorescence imaging is increased via a fluorophore caging procedure that uses a reducing agent to convert dyes to a long-lived dark state from which they can be activated with UV light and emit high numbers of photons.
- Joshua C Vaughan
- , Shu Jia
- & Xiaowei Zhuang
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Research Highlights |
More dyes enter the realm of nanoscopy
Widely used dyes for conventional microscopy of subcellular structures can also be used for super-resolution imaging.
- Daniel Evanko
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Correspondence |
Aptamers as potential tools for super-resolution microscopy
- Felipe Opazo
- , Matthew Levy
- & Silvio O Rizzoli
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News & Views |
Faster and more versatile tools for super-resolution fluorescence microscopy
Two methods for estimating fluorophore positions provide useful new options to researchers performing either single-particle tracking or super-resolution imaging.
- Alex Small
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Brief Communication |
Rational design of true monomeric and bright photoactivatable fluorescent proteins
Structure determination followed by targeted engineering of the popular photoactivatable fluorescent protein monomeric (m)Eos2 yields mEos3 versions that are more monomeric and less disruptive in protein fusions and also exhibit higher labeling density, brightness and other beneficial properties.
- Mingshu Zhang
- , Hao Chang
- & Tao Xu
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Brief Communication |
A simple, versatile method for GFP-based super-resolution microscopy via nanobodies
Nanobodies that bind to fluorescent proteins with high affinity and are coupled to bright organic dyes allow simple efficient labeling of fusion proteins for super-resolution microscopy.
- Jonas Ries
- , Charlotte Kaplan
- & Helge Ewers
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Correspondence |
Reply to "'Self-healing' dyes: intramolecular stabilization of organic fluorophores"
- Scott C Blanchard
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Methods in Brief |
Fast two-color structured-illumination microscopy
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Correspondence |
'Self-healing' dyes: intramolecular stabilization of organic fluorophores
- Philip Tinnefeld
- & Thorben Cordes
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News & Views |
Super resolution for common probes and common microscopes
A sophisticated analysis approach based on the concept of fluorophore localization provides dynamic super-resolution data of GFP-labeled live cells using a common, arc lamp–based wide-field fluorescence microscope.
- Keith A Lidke
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Brief Communication |
Dual-objective STORM reveals three-dimensional filament organization in the actin cytoskeleton
The use of dual-objective detection with astigmatism-based three-dimensional stochastic optical reconstruction microscopy (STORM) imaging improves resolution more than twofold and removes noise in resulting super-resolution images. This allowed detailed fluorescence imaging of distinctive features of the three-dimensional actin cytoskeleton ultrastructure with single-filament resolution in cells.
- Ke Xu
- , Hazen P Babcock
- & Xiaowei Zhuang
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Analysis |
Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging
A quantitative characterization of the switching properties of 26 organic dyes relates these properties to the quality of localization-based super-resolution images they generate. The data are a useful resource for selecting dyes and point to avenues for future analysis.
- Graham T Dempsey
- , Joshua C Vaughan
- & Xiaowei Zhuang
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News in Brief |
Deep imaging with STED
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Brief Communication |
Super-resolution 3D microscopy of live whole cells using structured illumination
Live-cell volumetric super-resolution imaging with 120-nm lateral and 360-nm axial resolution using structured-illumination microscopy at speeds of up to 5 s per cell volume over >50 time points captures fine cellular dynamics using only low illumination intensities.
- Lin Shao
- , Peter Kner
- & Mats G L Gustafsson
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Brief Communication |
Live-cell 3D super-resolution imaging in thick biological samples
The combination of light-sheet microscopy and localization-based super-resolution imaging allows deep subdiffraction resolution imaging in thick scattering specimens as demonstrated by three-dimensional super-resolution imaging of proteins in live 150-μm-diameter cell spheroids.
- Francesca Cella Zanacchi
- , Zeno Lavagnino
- & Alberto Diaspro
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Article |
Fast, three-dimensional super-resolution imaging of live cells
Judicious choice of probes and imaging conditions allows two-dimensional super-resolution imaging of live cells at speeds up to 2 Hz with ~25-nm resolution and three-dimensional super-resolution imaging at ~1 Hz with ~30 nm x-y and ~50 nm z dimension resolution using stochastic optical reconstruction microscopy (STORM).
- Sara A Jones
- , Sang-Hee Shim
- & Xiaowei Zhuang
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Correspondence |
DAOSTORM: an algorithm for high- density super-resolution microscopy
- Seamus J Holden
- , Stephan Uphoff
- & Achillefs N Kapanidis
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News & Views |
An in-depth view
Two fluorescence microscopy techniques provide alternative routes to obtaining three-dimensional super-resolution images with greater axial depth.
- Bo Huang
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Article |
Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores
Analyzing the first and higher-order moments of the diffraction spot of a 4Pi fluorescence detection scheme facilitates two-color, three-dimensional super-resolution microscopy with ~6 nm axial and ~8–23 nm lateral resolution in a layer ~650 nm thick.
- Daniel Aquino
- , Andreas Schönle
- & Alexander Egner
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Article |
Confined activation and subdiffractive localization enables whole-cell PALM with genetically expressed probes
Confined photoactivation of photoactivatable mCherry using two-photon illumination with line-scanning temporal focusing in combination with three-dimensional localization algorithms allows three-dimensional super-resolution microscopy of cellular features at <50 nm lateral and <100 nm axial resolution and depths greater than 8 μm.
- Andrew G York
- , Alireza Ghitani
- & Hari Shroff
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Correspondence |
Live-cell dSTORM with SNAP-tag fusion proteins
- Teresa Klein
- , Anna Löschberger
- & Markus Sauer
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Method to Watch |
Fast 3D super-resolution fluorescence microscopy
High-speed fluorescence imaging in all three dimensions at nanometer resolution will resolve, in finer detail, the workings of the living cell.
- Erika Pastrana
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Brief Communication |
Live-cell super-resolution imaging with trimethoprim conjugates
Use of a trimethoprim chemical tag allows super-resolution live-cell microscopy by stochastic single molecule–based localization imaging of the dynamics of genetically tagged histone H2B in cell nuclei.
- Richard Wombacher
- , Meike Heidbreder
- & Markus Sauer
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Brief Communication |
Optimized localization analysis for single-molecule tracking and super-resolution microscopy
A theoretical and experimental treatment of fitting methods for localizing the centers of diffraction-limited spots is presented. Use of an analytical point spread function shows that maximum likelihood fitting is superior to both unweighted and weighted least squares Gaussian fitting.
- Kim I Mortensen
- , L Stirling Churchman
- & Henrik Flyvbjerg