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  • Two-color STED Microscopy of Living Synapses using a Single Laser-beam Pair

    The advent of superresolution microscopy has opened up new research opportunities into dynamic processes at the nanoscale inside living biological specimens. This is particularly true for synapses, which are very small, highly dynamic, and embedded in brain tissue. Stimulated emission depletion (STED) microscopy, a recently developed laser-scanning technique, has been shown to be well suited for imaging living synapses in brain slices using yellow fluorescent protein as a single label. However, it would be highly desirable to be able to image presynaptic boutons and postsynaptic spines, which together form synapses, using two different fluorophores.
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  • Modern Fluorescent Proteins and their Biological Applications

    Here we present two review articles on fluorescent proteins and their biological applications. These first article reviews our current knowledge of blue, green, and red chromophore formation in permanently emitting FPs, photoactivatable FPs, and fluorescent timers. The second article focuses on novel monomeric RFPs and their application for studying gene expression, nuclear localization, and dynamics using advanced imaging.
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  • Webinar: Applications, Labeling Strategies and Fluorophores for Super-Resolution

    The free online webinar on super-resolution presented by Leica Microsystems in association with Microscopy & Analysis took place on Tuesday, 15 November 2011. Register and view the webinar on demand.
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  • Neurobiology and Microscopy

    Neurobiology, the science of nerves and the brain, has mainly been driven forward in the last 200 years by microscopic investigations. The structures of cellular and subcellular structures, interaction and the three-dimensional assembly of neurons were made visible by various microscopy techniques. The optical microscope is also a necessary tool for visualizing micropipettes in electrophysiological measurements. Thirdly, many types of functional imaging are performed by means of optical microscopy.
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  • Abstracts of the First European Super-Resolution User-Club Meeting

    The first European Super-resolution User-Club meeting took place from October 27 to 29 in Göttingen, Germany. Prof. Stefan Hell, the inventor of the STED technology, has hosted this first meeting. The user club is aimed at pioneering researchers from the European scientific community, who are early adopters and developers of super-resolution techniques.
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  • Natural Killer Cell Lytic Granule Secretion Occurs through a Pervasive Actin Network at the Immune Synapse

    Accumulation of filamentous actin (F-actin) at the immunological synapse (IS) is a prerequisite for the cytotoxic function of natural killer (NK) cells. Subsequent to reorganization of the actin network, lytic granules polarize to the IS where their contents are secreted directly toward a target cell, providing critical access to host defense.
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  • Webinar: Super-Resolution

    In 1873, Ernst Abbe developed a theory that defined the limit of resolution of the light microscope. Following suit from astronomy, Abbe defined resolution as the ability to resolve, as separate, two point sources of light. The Abbe limit of 200–300 nm is based upon the ability of the light microscope to collect only a subset of spatial frequencies and the physiological properties of the human eye.
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  • STED Nanoscopy of Actin Dynamics in Synapses deep inside Living Brain Slices

    It is difficult to investigate the mechanisms that mediate long-term changes in synapse function because synapses are small and deeply embedded inside brain tissue. Although recent fluorescence nanoscopy techniques afford improved resolution, they have so far been restricted to dissociated cells or tissue surfaces. However, to study synapses under realistic conditions, one must image several cell layers deep inside more-intact, three-dimensional preparations that exhibit strong light scattering, such as brain slices or brains in vivo.
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  • Unlimited Resolution - STED

    STED uses a differential method of two different diffraction patterns, where one pattern excites and the second pattern de-excites fluorochromes. The residual excited area is controllable by intensity down to (theoretically) zero – unlimited resolution.
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  • A STED-y Route to Commercialization

    The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken.
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  • Nanoscopy in a Living Multicellular Organism Expressing GFP

    We report superresolution fluorescence microscopy in an intact living organism, namely Caenorhabditis elegans nematodes expressing green fluorescent protein (GFP)-fusion proteins. We also superresolve, by stimulated emission depletion (STED) microscopy, living cultured cells, demonstrating that STED microscopy with GFP can be widely applied.
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  • The Missing Link to the Nanocosm of Life

    Fully understanding the functionality and complexity of the human central nervous system remains one of the major open questions in modern science. Stimulated emission depletion microscopy (STED) can be the method to reveal biological nanostructures
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  • Restless Receptors

    Synapses are the switch-points in our brain for information transmission, learning and memory. News studies and developments of imaging techniques have provided new insights into the dynamics of glutamate receptors. The use of superresolution technologies is making an essential contribution to this research.
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  • The Fate of Synaptic Vesicle Components upon Fusion

    Neurotransmitter release relies on the fusion of synaptic vesicles with the plasma membrane of synaptic boutons, which is followed by the recycling of vesicle components and formation of new vesicles. It is not yet clear whether upon fusion the vesicles persist as multimolecular patches in the plasma membrane, or whether they segregate into individual components.
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  • A Guide to Super-Resolution Fluorescence Microscopy

    For centuries, cell biology has been based on light microscopy and at the same time been limited by its optical resolution. However, several new technologies have been developed recently that bypass this limit.
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  • Confocal Nanoscopy Goes Multicolor

    Scientists strive to understand the architecture of life. They want to learn how biological structures are arranged in respect to one another. Multicolor superresolution imaging allows fundamental questions to be addressed by far-field fluorescence microscopy in unprecedented detail.
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  • Observing Life’s Nanostructures with STED

    The secrets of life and the causes of many diseases can only be fully explained if we understand the functions of the smallest components of organisms. Using the super high resolution STED microscope, research scientists are now able to observe cellular proteins and molecular structures measuring only a few nanometres.
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  • In Vivo Labeling Method Using a Genetic Construct for Nanoscale Resolution Microscopy

    We demonstrate beam scanning-stimulated emission depletion microscopy with in vivo labeled cells. A red emitting fluorescent dye is introduced into membrane protein fused to a multifunctional reporter protein (HaloTag, Promega, Madison, WI) in live cells. This approach allows superresolution stimulated emission depletion imaging without the limitations of immunofluorescence-based staining.
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  • Interview with Stefan Hell – For me, Pioneering is …

    Our interview partner on the topic of Pioneering is Prf. Stefan Hell, a scientific member of the Max Planck Society and director at the Max Planck Institute for Biophysical Chemistry in Göttingen, Germany, where he heads the Department of Nanobiophotonics. In addition to further chairs and memberships, he is also head of the High resolution Optical Microscopy research group, a partnership department of the German Cancer Research Center (DFKZ).
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