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| Open AccessQuantitative super-resolution imaging of Bruchpilot distinguishes active zone states
Complex molecular interactions occur in the active zone cytomatrix (CAZ) within the presynaptic terminal to regulate synaptic plasticity. Here, the authors use imaging techniques to show that the CAZ is composed of units containing on average 137 Bruchpilot proteins, many of which are arranged into clusters.
- Nadine Ehmann
- , Sebastian van de Linde
- & Robert J. Kittel
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Article
| Open AccessAlarmin S100A8/S100A9 as a biomarker for molecular imaging of local inflammatory activity
Alarmins are locally released during inflammation and are early amplifiers of inflammation. Here Vogl et al.show that the alarmin S100A8/S100A9 can be used as a sensitive marker to detect subclinical inflammation and follow disease progression in a variety of disease models.
- Thomas Vogl
- , Michel Eisenblätter
- & Johannes Roth
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Direct optical sensing of single unlabelled proteins and super-resolution imaging of their binding sites
The development of biosensors with improved sensitivity for the detection of biological molecules presents obvious possibilities for improved diagnostic tools in healthcare. Here, the authors present a technique for the label-free detection and super-resolution imaging of single proteins.
- Marek Piliarik
- & Vahid Sandoghdar
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Singlet oxygen triplet energy transfer-based imaging technology for mapping protein–protein proximity in intact cells
Fluorescence imaging can be used for the characterization of protein interactions in cells, although typical methods require the interacting groups to be quite close. Here, the authors present a fluorescent technique using singlet oxygen for the imaging of more distant protein–protein interactions.
- Tsz-Leung To
- , Michael J. Fadul
- & Xiaokun Shu
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Article
| Open AccessChemical reactivation of quenched fluorescent protein molecules enables resin-embedded fluorescence microimaging
Resin embedding of specimens is widely used for microscopy but can cause the loss of fluorescence from green-fluorescent protein. Here, the authors show that these proteins can be reactivated in resin-embedded samples through the use of alkaline buffer.
- Hanqing Xiong
- , Zhenqiao Zhou
- & Shaoqun Zeng
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Article
| Open AccessSeparating NADH and NADPH fluorescence in live cells and tissues using FLIM
NAD and NADP play fundamentally different roles in cellular metabolism, and yet these pyridine nucleotides cannot be distinguished spectroscopically in living cells. Blacker et al.demonstrate that fluorescence lifetime imaging can be used to quantify NADPH/NADH balance in cultured cells and in the mammalian cochlea.
- Thomas S. Blacker
- , Zoe F. Mann
- & Michael R. Duchen
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An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging
Infrared fluorescent proteins offer advantages for deep in vivo imaging thanks to the tissue-penetrating properties of infrared light. Here, Yu et al. design a monomeric infrared fluorescent protein that, when combined with expression of haeme oxygenase in cells, shows improved performance for in vivoimaging of neurons and brain tumours.
- Dan Yu
- , William Clay Gustafson
- & Xiaokun Shu
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Macromolecular structures probed by combining single-shot free-electron laser diffraction with synchrotron coherent X-ray imaging
Macromolecular complexes hold promise for future generations of drug delivery carriers, but probing their structures with high resolution is challenging. Here, the authors combine X-ray free-electron laser and synchrotron approaches to reveal the core-shell structure of RNA interference microsponges.
- Marcus Gallagher-Jones
- , Yoshitaka Bessho
- & Changyong Song
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A red-shifted fluorescent substrate for aldehyde dehydrogenase
Aldehyde dehydrogenase (ALDH)-expressing stem cells can be stained and sorted using a green fluorescent substrate. Minn et al.synthesise a red-shifted fluorescent ALDH probe and demonstrate sorting and microscopic staining of cells in the presence of a green fluorescent marker.
- Il Minn
- , Haofan Wang
- & Martin G. Pomper
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| Open AccessImaging of molecular surface dynamics in brain slices using single-particle tracking
Lateral diffusion of transmembrane signalling molecules is implicated in neuronal communication but imaging in tissue is limited by poor temporal resolution. Here, the authors use quantum dots to label lipids and adhesion molecules, allowing them to track single-molecule motions in subcellular compartments.
- B. Biermann
- , S. Sokoll
- & M. Heine
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Real-time in vivo imaging of invasive- and biomaterial-associated bacterial infections using fluorescently labelled vancomycin
Invasive and biomaterial-associated infections in humans are often difficult to diagnose and treat. Here, the authors demonstrate that real-time in vivodetection and imaging of these bacterial infections can be successfully performed with fluorescently labelled vancomycin.
- Marleen van Oosten
- , Tina Schäfer
- & Gooitzen M. van Dam
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Imaging energy status in live cells with a fluorescent biosensor of the intracellular ATP-to-ADP ratio
The ratio between ATP and ADP within the cell is a key indicator of metabolic status. Tantama et al. describe a ratiometric, genetically encoded fluorescent sensor for ATP:ADP that is now optimized for mammalian cells, and demonstrate that it can detect physiological changes in energy consumption and production.
- Mathew Tantama
- , Juan Ramón Martínez-François
- & Gary Yellen
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A peptide probe for targeted brown adipose tissue imaging
Current methods to image brown adipose tissue rely on radioactive tracers and specialized imaging equipment. Here Azhdarinia et al.report a peptide-based probe that selectively binds to the vascular endothelium of brown adipose tissue and allows the near-infrared imaging of brown fat in mice.
- Ali Azhdarinia
- , Alexes C. Daquinag
- & Mikhail G. Kolonin
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Article
| Open AccessVisualizing influenza virus infection in living mice
Mouse models of influenza infection that permit monitoring of infection in living animals are scarce. Here Pan et al. describe an engineered influenza virus expressing luciferase, which enables real-time in vivovisualization of viral infection and assessment of antiviral drugs.
- Weiqi Pan
- , Zhenyuan Dong
- & Ling Chen
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Non-invasive in vivo assessment of IDH1 mutational status in glioma
The metabolic reaction catalysed by the isocitrate dehydrogenase 1 (IDH1) enzyme is commonly perturbed in some glioma subtypes due to gain-of-function mutations in the IDH1 gene. Here, Chaumeil et al.present a method that detects mutant IDH1 activity by measuring the levels of different hyperpolarized metabolites produced by wild-type and mutant IDH1.
- Myriam M. Chaumeil
- , Peder E. Z. Larson
- & Sabrina M. Ronen
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In vivo imaging and histochemistry are combined in the cryosection labelling and intravital microscopy technique
Imaging cells within live animals is important in biomedical research, but this process is limited by the availability of probes. Ritsma et al.combine intravital microscopy with immunohistochemistry to increase the panel of available reagents and examine the role of T cells in the migration of breast cancer cells.
- Laila Ritsma
- , Nienke Vrisekoop
- & Jacco van Rheenen
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Article
| Open AccessDirect imaging of single UvrD helicase dynamics on long single-stranded DNA
Tracking single molecules on long stretches of single-stranded DNA poses technical challenges due to its propensity to form hairpin structures. To solve this problem, the authors combine TIRF microscopy with optical tweezers to stretch the DNA and capture the dynamics of DNA unwinding by UvrD DNA helicase.
- Kyung Suk Lee
- , Hamza Balci
- & Taekjip Ha
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Single-cell and subcellular pharmacokinetic imaging allows insight into drug action in vivo
Current pharmacokinetic models describe the distribution of drugs within tissues but usually lack single-cell resolution. Here Weissleder and colleagues visualize the subcellular distribution of an anticancer drug in real time in living animals and develop a model to extrapolate these findings to humans.
- Greg M. Thurber
- , Katy S. Yang
- & Ralph Weissleder
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Sequential then interactive processing of letters and words in the left fusiform gyrus
Reading requires accurate and rapid distinction between similar visual stimuli. Thesen and colleagues use human intracranial electrophysiology and brain imaging to show that letter-selective responses, in an area of the brain immediately posterior to the visual word-form area, occur before word selection.
- Thomas Thesen
- , Carrie R. McDonald
- & Eric Halgren
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| Open AccessLuminescent proteins for high-speed single-cell and whole-body imaging
Luminescent proteins are important tools for biomedical imaging but tend to emit fairly little light. Saito et al.. describe a brighter version of a bioluminescent protein that can visualize intracellular dynamics of various signalling molecules with high spatial and temporal resolution.
- Kenta Saito
- , Y-F Chang
- & Takeharu Nagai
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| Open AccessDynamic blastomere behaviour reflects human embryo ploidy by the four-cell stage
Abnormal human embryo development is implicated in the embryo arrest observed during in vitrofertilization. Chavez and colleagues perform time-lapse imaging on human embryos and find that chromosomally abnormal embryos exhibit diverse cell cycle parameters that may contribute to arrest.
- Shawn L. Chavez
- , Kevin E. Loewke
- & Renee A. Reijo Pera
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Imaging the post-fusion release and capture of a vesicle membrane protein
Rapid synaptic transmission requires efficient recycling of synaptic vesicle membrane proteins. Sochackiet al.use live cell, electron and super-resolution microscopy to visualize exocytosis of vesicular transporters and their rapid recapture in clathrin-rich microdomains in the plasma membrane.
- Kem A. Sochacki
- , Ben T. Larson
- & Justin W. Taraska
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Real-time in vivo imaging of the beating mouse heart at microscopic resolution
Microscopic imaging techniques have a high spatio-temporal resolution but, in living animals, are hampered by cardiac and respiratory motion. This paper describes a microscopic setup that allows fluorescent confocal imaging of the beating mouse heart over a period of several hours.
- Sungon Lee
- , Claudio Vinegoni
- & Ralph Weissleder
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Imaging enzyme-triggered self-assembly of small molecules inside live cells
Supramolecular interactions allow some small molecules to self-assemble into nanofibres and hydrogels in aqueous environments. Gaoet al.report a hydrogelator that forms fluorescent nanofibres within cells, leading to the visualization of their self-assembly at the endoplasmic reticulum.
- Yuan Gao
- , Junfeng Shi
- & Bing Xu
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| Open AccessDeep-tissue focal fluorescence imaging with digitally time-reversed ultrasound-encoded light
Fluorescence imaging is important for biomedical research and applications, but the turbidity of biological material prohibits deep tissue study. By combining ultrasound-encoding with digital time-reversal, Wanget al.perform focussed fluorescence imaging at a tissue depth of 2.5mm.
- Ying Min Wang
- , Benjamin Judkewitz
- & Changhuei Yang
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Article
| Open AccessNeural circuits in the brain that are activated when mitigating criminal sentences
Jurors can be influenced by mitigating circumstances when deciding on sentences for committed crimes. Yamadaet al. show that feelings of sympathy created by mitigating circumstances activate moral conflict regions of the brain that predict individual differences in the severity of the sentence.
- Makiko Yamada
- , Colin F. Camerer
- & Hidehiko Takahashi
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Article
| Open AccessIntracellular temperature mapping with a fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy
Intracellular temperature mapping has not previously been achieved. Now, a fluorescent polymeric thermometer has been developed that can be used in combination with fluorescence-lifetime imaging microscopy to allow thermometry with spatial and temperature resolutions of 200 nm and 0.18–0.58 ° C.
- Kohki Okabe
- , Noriko Inada
- & Seiichi Uchiyama
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Biomedical photoacoustics beyond thermal expansion using triggered nanodroplet vaporization for contrast-enhanced imaging
A dual-contrast agent has been developed for combined ultrasound and photoacoustic imaging. This agent uses vaporization for ultrasound contrast enhancement and photoacoustic signal generation, providing significantly higher signals than thermal expansion, the most commonly used photoacoustic mechanism.
- Katheryne Wilson
- , Kimberly Homan
- & Stanislav Emelianov
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Super-resolution surface mapping using the trajectories of molecular probes
Surface characterization of soft materialsin situis challenging due to the importance of non-covalent interactions. Now, a new chemical imaging method is reported that generates images of surface interactions by combining many molecular probe trajectories.
- Robert Walder
- , Nathaniel Nelson
- & Daniel K. Schwartz
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A synthetic icosahedral DNA-based host–cargo complex for functional in vivo imaging
Encapsulating molecules within supramolecular frameworks for potential biological application is challenging. Bhatiaet al. incorporate a fluorescent polymer within an icosahedral DNA nanocapsule, and show that it can be used to target specific cells in vivoand map pH spatially and temporally.
- Dhiraj Bhatia
- , Sunaina Surana
- & Yamuna Krishnan
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Subcellular in vivo time-lapse imaging and optical manipulation of Caenorhabditis elegans in standard multiwell plates
The nematodeCaenorhabditis elegans is a widely used model organism for visualizing development, ageing and other phenomena, but high-throughput imaging requires immobilization of the animal. Rohde and Yanik report a rapid immobilization and imaging method for the high-throughput screening of C. elegans.
- Christopher B. Rohde
- & Mehmet Fatih Yanik
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Development and host cell modifications of Plasmodium falciparum blood stages in four dimensions
ThePlasmodium falciparum parasite that causes malaria has a complex life cycle in human erythrocytes. Using time-lapse three-dimensional imaging, the authors show the intraerythrocytic stages of the parasite and provide new insight into the export of P. falciparumproteins to Maurer's clefts.
- Christof Grüring
- , Arlett Heiber
- & Tobias Spielmann
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Image transmission through an opaque material
The challenge of reconstructing the image of an object when viewed through an opaque material is of particular importance for biological tissues. Here, the authors show that it is possible to reconstruct the image of a complex object from interference patterns of multiple wavefronts using phase-shifting interferometry.
- Sébastien Popoff
- , Geoffroy Lerosey
- & Sylvain Gigan
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