Download or read book Nanoscale Imaging of Synapses written by U. Valentin Nägerl and published by Humana. This book was released on 2016-08-23 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synapses underlie rapid and flexible neural communication in the brain and they hold the key to understanding higher brain functions in health and disease. Because they are very small and highly dynamic, it is very difficult to study them with traditional techniques. Fortunately, recent ground-breaking advances in optical microscopy (e.g. STED, PALM, STORM, SIM) have greatly improved our ability to image living synapses at the nanoscale, even down to the level of single molecules. The proposed volume brings together leading researchers to review these exciting new techniques and their application in neurobiological research. It will explain and discuss the basic principles behind the various superresolution modalities, how they are implemented, what their scope and limitations are etc. In addition, several key research discoveries on synapses enabled by these novel approaches will be highlighted.

Download or read book Nanoscale Imaging of Synapse Morphology in the Mouse Neocortex in Vivo by Two photon STED Microscopy written by Mirelle Jamilla Tamara Ter Veer and published by . This book was released on 2016 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The brain is a complex organ consisting of neurons and non-neuronal cells. Communication between neurons takes place via synapses, whose morphological remodeling is thought to be crucial for information processing and storage in the mammalian brain. Recently, this neuro-centric view of synaptic function has evolved, also taking into account the glial processes in close vicinity of the synapse. However, as their structure is well below the spatial resolution of conventional light microscopy, progress in investigating them in a physiological environment, the intact brain, has been impeded. Indeed, little is known on the nanoscale morphological variations of dendritic spines, the interaction with glial processes, and how these affect synaptic transmission in vivo. Here, we aim to visualize the dynamic nano-morphology of dendritic spines in mouse somatosensory cortex in vivo. We implemented super-resolution 2P-STED time-lapse imaging, which allows for high spatial resolution and deep tissue penetration, in anesthetized mice, and show that the nano-morphology of spines is diverse, variable, but on average stable, and that differences in spine morphology can have an effect on spine biochemical compartmentalization in vivo. Moreover, implementation of dual color in vivo super-resolution imaging and a novel astrocytic labeling approach provided the first steps towards nanoscale characterization of neuron-glia interactions in vivo. These findings bring new insights in synapse dynamics at the nanoscale in vivo, and our methodological endeavors help pave the way for a better understanding of how nanoscale aspects of spine morphology and their dynamics might contribute to brain physiology and animal behavior.

Download or read book Quantitative Nanoscale Imaging of Synaptic Protein Organization written by Lara Laparra Cuervo and published by . This book was released on 2019 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: The arrival of super-resolution techniques has driven researchers to explore biological areas that were unreachable before. Such techniques not only allowed the improvement of spatial resolution in images but also the possibility to perform quantitative measurements at the single-molecule level. The interest in that particular field has been growing over the years and new and more sophisticated tools have been developed. Neuroscience has been one of the first fields to adapt and benefit from super-resolution microscopy. These techniques opened a new window of opportunity to reveal spatial organization of the neuronal cytoskeleton and the molecular organization and dynamics of the synapse, structures below the spatial resolution limit of conventional light microscopy. Protein organization and stoichiometry is central for synaptic transmission in neurons. Knowing the absolute numbers of proteins playing a key role in diseases can be of extreme interest in order to understand the mechanisms that lead to such neurological disorders. In this thesis we exploited the super-resolution techniques to develop a pioneering method for quantitative single-molecule measurements and to unravel the organization of a synaptic protein complex that was never visualized before with nanoscale spatial resolution. We developed a novel method in order to quantify the photoactivation efficiency of eight different photoswitchable fluorescent proteins commonly used in super-resolution experiments. We used the glycine receptor as a template because of its well-known stoichiometry and tagged eight different photoswitchable fluorescent proteins to the a- and ß-subunits of this receptor and transiently transfected them to Xenopus oocytes. The fact that the fluorescent proteins are genetically encoded make them highly suitable for quantitative single-molecule counting. The photoactivation efficiency, which is the percentage of a fluorescent protein that photoactivates into a fluorescently detectable form, plays a critical role in properly interpreting quantitative measurements. Moreover, we also focused our studies on super-resolution imaging of a synaptic protein complex, called LGI1 complex. This ensemble of proteins is one of the main key players involved in different neurological disorders. Leucine rich glioma activated 1 (LGI1) is a neuronal protein that forms a trans-synaptic bridge linking pre- and postsynaptic transmembrane proteins (ADAM22 and ADAM23) and helps to organize a multimeric complex at the synapse including AMPA receptors and voltage-gated potassium channels (VGKC). LGI1 autoimmune encephalitis is a severe neuropsychiatric disorder related to epilepsy where the patients produce autoantibodies against LGI1, which alter synaptic plasticity. However, the molecular mechanisms that lead to the observed problems in patients still remain largely unknown. Using well-characterized synaptic markers as molecular standards, we determined the positioning of LGI1 and the other related proteins within the synaptic space at nanoscale resolution by means of multi-color STORM. Further, the comparison of this molecular architecture in healthy neurons versus neurons treated with antibodies from patients suffering from LGI1 autoimmune encephalitis showed that these antibodies impact the nanoscale organization of pre-synaptic proteins. These results suggested a loss of LGI1 interaction with pre-synaptic proteins upon antibody binding and gave further insight into early changes in pathology.

Download or read book Quantifying and Controlling the Nano Architecture of Neuronal Synapses written by Xiaobing Chen and published by Frontiers Media SA. This book was released on 2022-09-27 with total page 195 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Far Field Optical Nanoscopy written by Philip Tinnefeld and published by Springer. This book was released on 2015-02-07 with total page 340 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book describes developments in the field of super-resolution fluorescence microscopy or nanoscopy. In 11 chapters, distinguished scientists and leaders in their respective fields describe different nanoscopy approaches, various labeling technologies, and concrete applications. The topics covered include the principles and applications of the most popular nanoscopy techniques STED and (f)PALM/STORM, along with advances brought about by fluorescent proteins and organic dyes optimized for fluorescence nanoscopy. Furthermore, the photophysics of fluorescent labels is addressed, specifically for improving their photoswitching capabilities. Important applications are also discussed, such as the tracking and counting of molecules to determine acting forces in cells, and quantitative cellular imaging, respectively, as well as the mapping of chemical reaction centers at the nano-scale. The 2014 Chemistry Nobel Prize® was awarded for the ground-breaking developments of super-resolved fluorescence microscopy. In this book, which was co-edited by one of the prize winners, readers will find the most recent developments in this field.

Download or read book Expansion Microscopy of C Elegans written by Chih-Chieh Jay Yu and published by . This book was released on 2020 with total page 146 pages. Available in PDF, EPUB and Kindle. Book excerpt: Expansion microscopy (ExM) enables 3-D, nanoscale-precise imaging of biological specimens by isotropic swelling of hydrogel-embedded, chemically processed tissue. Such capability raises the question of whether nanoscale mapping of biomolecules could be performed in an entire organism, which would allow super-resolution-mediated in situ analyses, such as digital quantification of biomolecules and mapping of synaptic contacts, to be performed within the context of an entire nervous system. The nematode Caenorhabditis elegans could be a suitable model for such organism-wide analyses, due to its tractable physical size, deterministic cell lineage, ease of genetic control, and well-established literature. However, C. elegans is enclosed in a chemically impermeable and mechanically tough cuticle, which could hinder the deployment of ExM. In this thesis, we present a strategy, expansion of C. elegans (ExCel), to expand fixed, cuticle-enclosed intact animals of C. elegans. ExCel enables simultaneous readout of fluorescent proteins, RNAs, DNA locations, and anatomical structures at resolutions of ~65-75 nm (3.3-3.8x linear expansion). We also developed epitope-preserving ExCel, which enables imaging of endogenous proteins stained by antibodies, and iterative ExCel, which enables imaging of fluorescent proteins at a ~25-nm resolution (20x linear expansion). We demonstrate the utility of the ExCel toolbox for multiplexed imaging of multiple molecular types, for mapping synaptic proteins, for identifying previously unreported proteins at cell junctions, and for gene expression analysis in multiple individual neurons of the same animal. In addition to ExCel, we discuss two other ExM-related technologies, including tetragel, which is a highly homogeneous hydrogel network that improves the nanoscale isotropy of biological ultrastructure expanded by ExM, and stochastic arrangement of reporters in clusters (STARC), which is a strategy for recording neuronal activity at a subneurite-level resolution, in densely labeled neuronal populations. Taken together, the work presented in this thesis extends the capabilities of ExM, and lays the foundation for a comprehensive, functionally and structurally informed analysis of an entire organism, which could reveal new insights in neuroscience, organismal development, and systems biology.

Download or read book STED Nanoscopy of Synaptic Substructures in Living Mice written by Jennifer-Magdalena Masch and published by . This book was released on 2017 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Optical nanoscopy has revolutionized far-field microscopy, enabling the observation of subcellular structures and dynamics from completely new perspectives. Among other fields, neuroscience benefits greatly from the remarkable advances in super-resolution microscopy, which provide unprecedented insights into the molecular organization and function of synapses. STED nanoscopy has been one of the most successful methods for live-cell applications and is the only superresolution technique that has been demonstrated for imaging in living mice so far. However, in vivo nanoscale imaging of synapt...

Download or read book Single Molecule Microscopy in Neurobiology written by Nobuhiko Yamamoto and published by Humana. This book was released on 2021-05-13 with total page 331 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume looks at the methodology and techniques used by experts to study how certain molecules function in specific locations, and their temporal patterns. Chapters in this book cover topics such as in vivo single-molecule tracking of voltage-gated calcium channels with split-fluorescent proteins in CRISPR-engineering C. elegans; protein-protein interactions in membranes using single particle tracking; neuropathological diseases revealed by quantum-dot single particle tracking; SPoD-OnSPAN; and investigating molecular diffusion inside small neuronal compartments with two-photon fluorescence correlation spectroscopy. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and comprehensive, Single Molecule Microscopy is a valuable resource for any researcher interested in learning more about this important field.

Download or read book Synapse Development and Maturation written by Pasko Rakic and published by Academic Press. This book was released on 2020-05-29 with total page 560 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synapse Development and Maturation, the latest release in the Comprehensive Developmental Neuroscience series, presents the latest information on the genetic, molecular and cellular mechanisms of neural development. The book provides a much-needed update that underscores the latest research in this rapidly evolving field, with new section editors discussing the technological advances that are enabling the pursuit of new research on brain development. This volume focuses on the synaptogenesis and developmental sequences in the maturation of intrinsic and synapse-driven patterns. - Features leading experts in various subfields as section editors and article authors - Presents articles that have been peer reviewed to ensure accuracy, thoroughness and scholarship - Includes coverage of mechanisms which regulate synapse formation and maintenance during development - Covers neural activity, from cell-intrinsic maturation, to early correlated patterns of activity

Download or read book Pharmacology of Neurotransmitter Release written by Thomas C. Südhof and published by Springer Science & Business Media. This book was released on 2007-12-07 with total page 582 pages. Available in PDF, EPUB and Kindle. Book excerpt: It has been known for half a century that neurotransmitters are released in preformed quanta, that the quanta represent transmitter-storing vesicles, and that release occurs by exocytosis. The focus of this book is twofold. In the first part, the molecular events of exocytosis are analysed. In the second part of the book, the presynaptic receptors for endogenous chemical signals are presented that make neurotransmitter release a highly regulated process.

Download or read book Visualizing Chemistry written by National Research Council and published by National Academies Press. This book was released on 2006-06-01 with total page 222 pages. Available in PDF, EPUB and Kindle. Book excerpt: Scientists and engineers have long relied on the power of imaging techniques to help see objects invisible to the naked eye, and thus, to advance scientific knowledge. These experts are constantly pushing the limits of technology in pursuit of chemical imagingâ€"the ability to visualize molecular structures and chemical composition in time and space as actual events unfoldâ€"from the smallest dimension of a biological system to the widest expanse of a distant galaxy. Chemical imaging has a variety of applications for almost every facet of our daily lives, ranging from medical diagnosis and treatment to the study and design of material properties in new products. In addition to highlighting advances in chemical imaging that could have the greatest impact on critical problems in science and technology, Visualizing Chemistry reviews the current state of chemical imaging technology, identifies promising future developments and their applications, and suggests a research and educational agenda to enable breakthrough improvements.

Download or read book Nanoscale Co organization of AMPAR and Neuroligin Probed with Single molecule Based Microscopy written by Kalina Haas and published by . This book was released on 2013 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The brain is made of complex networks of interconnected neuronal cells. All our mental activities are underlain by electrochemical signals passing through dedicated neuronal circuits. Climbing further up on the complexity ladder, information processing by neurons is performed by multiple molecules assembling and interacting together. It is well accepted that the understanding of the molecular structuring inside neuronal cells is essential to apprehend functioning of the brain. For this reason, study of the organization of the key neuronal and synaptic molecules greatly contributes to understand the mystery of the brain. AMPA receptors (AMPARs) are ionotropic glutamate receptors that play a central role in synaptic plasticity and basal synaptic transmission in the central nervous system. The distribution of AMPARs on the neuronal membrane is remarkably heterogeneous. They are organized in distinct functional aggregates, called nanodomains. Previous work demonstrate that the postsynaptic adhesion molecule Neuroligin (NRLG) anchors AMPARs through PSD-95 in the postsynaptic membrane while simultaneously forming a trans-synaptic adhesion complex with presynaptic Neurexin (NRX), and recruiting vesicular release machinery at the presynaptic site. In this way, NRLG functionally organizes synapses by recruiting post and pre-synaptic molecules essential for regulation of synaptic responses. Here we studied the effect of NRLG modulation (modification of expression level or activity) on AMPAR nano-dynamics and nano-organization at individual synapses. Our hypothesis is that the NRX-NRLG complex could be involved in the precise localization of postsynaptic receptors and their apposition with the neurotransmitter release sites in the presynaptic active zone, thus playing important role in proper signal transduction. The size of the postsynaptic density (PSD) is in the order of 500 nm, whereas the average diameter of AMPAR nanodomains 100 nm. Such small dimension necessitated the application of super-resolution microscopy techniques, whose resolution in the range of 20-40 nm is almost an order of magnitude better than diffraction limited fluorescence microscopy. Probe-based far-field fluorescence nanoscopies allow visualizing cells down to almost molecular level. To achieve my goals, I implemented different single-molecule localization nanoscopies which rely on the detection of selected populations of fluorescence probes that are separated in space and time. It was proposed that membrane trafficking of neurotransmitter receptors may contribute to modulation of synaptic efficacy. I have probed diffusional properties of AMPARs with single particle tracking, which has long been applied to probe heterogeneity of the cell membrane. Relative localization of biomolecules provides the basis for understanding their functional relationship. It is well accepted that the juxtaposition of two objects, as well as their colocalization, may give evidence of their association. With the recent developments in multi-color acquisition of single molecule and ensemble based super resolution images, it is now possible to explore the colocalization at the nanoscale between biomolecules in live and fixed cells. Despite the popularity and wide spread application of super resolution imaging, there exist only a few quantitative analysis paradigms for the colocalization of multicolor super-resolution images. Here, with the aid of conventional colocalization measurement paradigms and multivariate statistics, we analyze and report in detail the scale segregation and proximity of macromolecules within functional zones of synapses. Furthermore, we use these paradigms to evaluate fluorescent tags involved in the routine generation of single molecule based super-resolution images. We extend our analysis to elucidate in depth the co-aggregation and clustering of two key synaptic molecules, PSD95 and AMPARs, which are involved in basal synaptic organization and transmission.

Download or read book Fluorescent Nanodiamonds written by Huan-Cheng Chang and published by John Wiley & Sons. This book was released on 2018-11-12 with total page 294 pages. Available in PDF, EPUB and Kindle. Book excerpt: The most comprehensive reference on fluorescent nanodiamond physical and chemical properties and contemporary applications Fluorescent nanodiamonds (FNDs) have drawn a great deal of attention over the past several years, and their applications and development potential are proving to be manifold and vast. The first and only book of its kind, Fluorescent Nanodiamonds is a comprehensive guide to the basic science and technical information needed to fully understand the fundamentals of FNDs and their potential applications across an array of domains. In demonstrating the importance of FNDs in biological applications, the authors bring together all relevant chemistry, physics, materials science and biology. Nanodiamonds are produced by powerful cataclysmic events such as explosions, volcanic eruptions and meteorite impacts. They also can be created in the lab by high-pressure high-temperature treatment of graphite or detonating an explosive in a reactor vessel. A single imperfection can give a nanodiamond a specific, isolated color center which allows it to function as a single, trapped atom. Much smaller than the thickness of a human hair, a nanodiamond can have a huge surface area that allows it to bond with a variety of other materials. Because of their non-toxicity, nanodiamonds may be useful in biomedical applications, such as drug delivery and gene therapy. The most comprehensive reference on a topic of rapidly increasing interest among academic and industrial researchers across an array of fields Includes numerous case studies and practical examples from many areas of research and industrial applications, as well as fascinating and instructive historical perspectives Each chapter addresses, in-depth, a single integral topic including the fundamental properties, synthesis, mechanisms and functionalisation of FNDs The first book published by the key patent holder with his research group in the field of FNDs Fluorescent Nanodiamonds is an important working resource for a broad range of scientists and engineers in industry and academia. It will also be a welcome reference for instructors in chemistry, physics, materials science, biology and related fields.

Download or read book Nanoscale Photonic Imaging written by Tim Salditt and published by Springer Nature. This book was released on 2020-06-09 with total page 634 pages. Available in PDF, EPUB and Kindle. Book excerpt: This open access book, edited and authored by a team of world-leading researchers, provides a broad overview of advanced photonic methods for nanoscale visualization, as well as describing a range of fascinating in-depth studies. Introductory chapters cover the most relevant physics and basic methods that young researchers need to master in order to work effectively in the field of nanoscale photonic imaging, from physical first principles, to instrumentation, to mathematical foundations of imaging and data analysis. Subsequent chapters demonstrate how these cutting edge methods are applied to a variety of systems, including complex fluids and biomolecular systems, for visualizing their structure and dynamics, in space and on timescales extending over many orders of magnitude down to the femtosecond range. Progress in nanoscale photonic imaging in Göttingen has been the sum total of more than a decade of work by a wide range of scientists and mathematicians across disciplines, working together in a vibrant collaboration of a kind rarely matched. This volume presents the highlights of their research achievements and serves as a record of the unique and remarkable constellation of contributors, as well as looking ahead at the future prospects in this field. It will serve not only as a useful reference for experienced researchers but also as a valuable point of entry for newcomers.

Download or read book Receptor and Ion Channel Detection in the Brain written by Rafael Luján and published by Humana. This book was released on 2016-02-02 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Receptor and Ion Channel Detection in the Brain provides state-of-the-art and up-to-date methodological information on molecular, neuroanatomical and functional techniques that are currently used to study neurotransmitter receptors and ion channels in the brain. The chapters have been contributed by world-wide recognized neuroscientists who explain in an easy and detailed way well established and tested protocols embracing molecular, cellular, subcellular, anatomical and electrophysiological aspects of the brain. This comprehensive and practical manual is presented in a simple, step-by-step manner for laboratory use, and also offers unambiguous detail and key implementation advice that proves essential for successful results and facilitate choosing the best method for the target proteins under study. This work serves as a useful guide for young researchers and students in training as well as for neurologists and established scientists who wish to extend their repertoire of techniques.

Download or read book Inorganic Nanoparticles written by Claudia Altavilla and published by CRC Press. This book was released on 2017-12-19 with total page 885 pages. Available in PDF, EPUB and Kindle. Book excerpt: Among the various nanomaterials, inorganic nanoparticles are extremely important in modern technologies. They can be easily and cheaply synthesized and mass produced, and for this reason, they can also be more readily integrated into applications. Inorganic Nanoparticles: Synthesis, Applications, and Perspectives presents an overview of these special materials and explores the myriad ways in which they are used. It addresses a wide range of topics, including: Application of nanoparticles in magnetic storage media Use of metal and oxide nanoparticles to improve performance of oxide thin films as conducting media in commercial gas and vapor sensors Advances in semiconductors for light-emitting devices and other areas related to the energy sector, such as solar energy and energy storage devices (fuel cells, rechargeable batteries, etc.) The expanding role of nanosized particles in the field of catalysis, art conservation, and biomedicine The book’s contributors address the growing global interest in the application of inorganic nanoparticles in various technological sectors. Discussing advances in materials, device fabrication, and large-scale production—all of which are urgently required to reduce global energy demands—they cover innovations in areas such as solid-state lighting, detailing how it still offers higher efficiency but higher costs, compared to conventional lighting. They also address the impact of nanotechnology in the biomedical field, focusing on topics such as quantum dots for bioimaging, nanoparticle-based cancer therapy, drug delivery, antibacterial agents, and more. Fills the informational gap on the wide range of applications for inorganic nanoparticles in areas including biomedicine, electronics, storage media, conservation of cultural heritage, optics, textiles, and cosmetics Assembling work from an array of experts at the top of their respective fields, this book delivers a useful analysis of the vast scope of existing and potential applications for inorganic nanoparticles. Versatile as either a professional research resource or textbook, this effective tool elucidates fundamentals and current advances associated with design, characterization, and application development of this promising and ever-evolving device.

Download or read book Super Resolution Imaging in Biomedicine written by Alberto Diaspro and published by CRC Press. This book was released on 2016-11-03 with total page 448 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book encompasses the full breadth of the super-resolution imaging field, representing modern techniques that exceed the traditional diffraction limit, thereby opening up new applications in biomedicine. It shows readers how to use the new tools to increase resolution in sub-nanometer-scale images of living cells and tissue, which leads to new information about molecules, pathways and dynamics. The book highlights the advantages and disadvantages of the techniques, and gives state-of-the-art examples of applications using microscopes currently available on the market. It covers key techniques such as stimulated emission depletion (STED), structured illumination microscopy (SSIM), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM). It will be a useful reference for biomedical researchers who want to work with super-resolution imaging, learn the proper technique for their application, and simultaneously obtain a solid footing in other techniques.