49,708 research outputs found
Image informatics strategies for deciphering neuronal network connectivity
Brain function relies on an intricate network of highly dynamic neuronal connections that rewires dramatically under the impulse of various external cues and pathological conditions. Among the neuronal structures that show morphologi- cal plasticity are neurites, synapses, dendritic spines and even nuclei. This structural remodelling is directly connected with functional changes such as intercellular com- munication and the associated calcium-bursting behaviour. In vitro cultured neu- ronal networks are valuable models for studying these morpho-functional changes. Owing to the automation and standardisation of both image acquisition and image analysis, it has become possible to extract statistically relevant readout from such networks. Here, we focus on the current state-of-the-art in image informatics that enables quantitative microscopic interrogation of neuronal networks. We describe the major correlates of neuronal connectivity and present workflows for analysing them. Finally, we provide an outlook on the challenges that remain to be addressed, and discuss how imaging algorithms can be extended beyond in vitro imaging studies
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Atomic electron tomography in three and four dimensions
Atomic electron tomography (AET) has become a powerful tool for atomic-scale structural characterization in three and four dimensions. It provides the ability to correlate structures and properties of materials at the single-atom level. With recent advances in data acquisition methods, iterative three-dimensional (3D) reconstruction algorithms, and post-processing methods, AET can now determine 3D atomic coordinates and chemical species with sub-Angstrom precision, and reveal their atomic-scale time evolution during dynamical processes. Here, we review the recent experimental and algorithmic developments of AET and highlight several groundbreaking experiments, which include pinpointing the 3D atom positions and chemical order/disorder in technologically relevant materials and capturing how atoms rearrange during early nucleation at four-dimensional atomic resolution
Wide-Field Multi-Parameter FLIM: Long-Term Minimal Invasive Observation of Proteins in Living Cells.
Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes
Real-Time analysis and visualization for single-molecule based super-resolution microscopy
Accurate multidimensional localization of isolated fluorescent emitters is a time consuming process in single-molecule based super-resolution microscopy. We demonstrate a functional method for real-time reconstruction with automatic feedback control, without compromising the localization accuracy. Compatible with high frame rates of EM-CCD cameras, it relies on a wavelet segmentation algorithm, together with a mix of CPU/GPU implementation. A combination with Gaussian fitting allows direct access to 3D localization. Automatic feedback control ensures optimal molecule density throughout the acquisition process. With this method, we significantly improve the efficiency and feasibility of localization-based super-resolution microscopy
Optimized imaging using non-rigid registration
The extraordinary improvements of modern imaging devices offer access to data
with unprecedented information content. However, widely used image processing
methodologies fall far short of exploiting the full breadth of information
offered by numerous types of scanning probe, optical, and electron
microscopies. In many applications, it is necessary to keep measurement
intensities below a desired threshold. We propose a methodology for extracting
an increased level of information by processing a series of data sets
suffering, in particular, from high degree of spatial uncertainty caused by
complex multiscale motion during the acquisition process. An important role is
played by a nonrigid pixel-wise registration method that can cope with low
signal-to-noise ratios. This is accompanied by formulating objective quality
measures which replace human intervention and visual inspection in the
processing chain. Scanning transmission electron microscopy of siliceous
zeolite material exhibits the above-mentioned obstructions and therefore serves
as orientation and a test of our procedures
Plasmonic antennas and zero mode waveguides to enhance single molecule fluorescence detection and fluorescence correlation spectroscopy towards physiological concentrations
Single-molecule approaches to biology offer a powerful new vision to
elucidate the mechanisms that underpin the functioning of living cells.
However, conventional optical single molecule spectroscopy techniques such as
F\"orster fluorescence resonance energy transfer (FRET) or fluorescence
correlation spectroscopy (FCS) are limited by diffraction to the nanomolar
concentration range, far below the physiological micromolar concentration range
where most biological reaction occur. To breach the diffraction limit, zero
mode waveguides and plasmonic antennas exploit the surface plasmon resonances
to confine and enhance light down to the nanometre scale. The ability of
plasmonics to achieve extreme light concentration unlocks an enormous potential
to enhance fluorescence detection, FRET and FCS. Single molecule spectroscopy
techniques greatly benefit from zero mode waveguides and plasmonic antennas to
enter a new dimension of molecular concentration reaching physiological
conditions. The application of nano-optics to biological problems with FRET and
FCS is an emerging and exciting field, and is promising to reveal new insights
on biological functions and dynamics.Comment: WIREs Nanomed Nanobiotechnol 201
R&D Paths of Pixel Detectors for Vertex Tracking and Radiation Imaging
This report reviews current trends in the R&D of semiconductor pixellated
sensors for vertex tracking and radiation imaging. It identifies requirements
of future HEP experiments at colliders, needed technological breakthroughs and
highlights the relation to radiation detection and imaging applications in
other fields of science.Comment: 17 pages, 2 figures, submitted to the European Strategy Preparatory
Grou
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