Leica Science Lab - Tag : Synapse https://www.leica-microsystems.com//science-lab/tag/tags/synapse/show/Tag/ Article tagged with Synapse en-US https://www.leica-microsystems.com/25022 EM Sample Preparation Bridging Structure and Dynamics at the Nanoscale through Optogenetics and Electrical Stimulation Nanoscale ultrastructural information is typically obtained by means of static imaging of a fixed and processed specimen. However, this is only a snapshot of one moment within a dynamic system in which structures are constantly changing. Exploring specific time points of a dynamic process is therefore a major challenge. Exploring a process at the nanoscale through optogenetics or electrical field stimulation in combination with timed millisecond precision vitrification is a promising technology to overcome this challenge. In the first part of a series of application notes the practical considerations of stimulation-assisted vitrification are discussed. https://www.leica-microsystems.com//science-lab/bridging-structure-and-dynamics-at-the-nanoscale-through-optogenetics-and-electrical-stimulation/ Mon, 20 May 2019 22:00:00 +0000 Dr. Andres Kaech, PhD Frédéric Leroux https://www.leica-microsystems.com/19872 Super-Resolution Abstracts of the 7th European Super-Resolution User-Club Meeting The 7th Super-Resolution User Club Meeting was held in collaboration with Prof Pavel Hozák , at the Institute of Molecular Genetics of the ASCR in Prague. Keeping the event close to science is one of the founding principles of the event, allowing all participants to network, share and explore exciting new super-resolution and nanoscopy applications. Central to this are the scientific talks given during the meeting, with this cutting-edge microscopy technique as their central theme. A wide selection of topics were covered, prompting interesting discussions during the workshops. https://www.leica-microsystems.com//science-lab/abstracts-of-the-7th-european-super-resolution-user-club-meeting/ Sun, 17 Dec 2017 23:00:00 +0000 Prof. DrSc. Pavel Hozák, Dr. Jindřiška Fišerová, Dr. Eugene Katrukha, Dr. Susan Cox, Prof. Christian Eggeling, Ph.D. Marc van Zandvoort, Dr. Jordi Andilla, Dr. Martin Offterdinger, Prof. Dr. Torsten Ochsenreiter, Dr. Camille Boutin https://www.leica-microsystems.com/19607 EM Sample Preparation Interview with Dr. Shigeki Watanabe on Research in Synaptic Membrane Dynamics Dr. Shigeki Watanabe, principle investigator of the department of Cell Biology at the Johns Hopkins University School of Medicine in Baltimore, held a workshop in Zürich, Switzerland on methods to study synaptic dynamics with millisecond precision. In collaboration with Dr. Andres Käch from the University of Zurich all workshop attendees enjoyed presentations and hands-on sessions on the EM ICE by Leica Microsystems with Light and Electrical Stimulation, revealing the latest developments in brain research. During this workshop Dr. Bernd Sägmüller from Leica Microsystems had the chance for an interview with Dr. Watanabe. https://www.leica-microsystems.com//science-lab/interview-with-dr-shigeki-watanabe-on-research-in-synaptic-membrane-dynamics/ Thu, 06 Jul 2017 23:00:00 +0000 Dr. Bernd Sägmüller, PhD Shigeki Watanabe https://www.leica-microsystems.com/19109 Confocal Microscopy Human NK Cell Development Requires CD56-mediated Motility and Formation of the Developmental Synapse While distinct stages of natural killer (NK) cell development have been defined, the molecular interactions that shape human NK cell maturation are poorly understood. Here we define intercellular interactions between developing NK cells and stromal cells which, through contact-dependent mechanisms, promote the generation of mature, functional human NK cells from CD34+ precursors. We show that developing NK cells undergo unique, developmental stage-specific sustained and transient interactions with developmentally supportive stromal cells, and that the relative motility of NK cells increases as they move through development in vitro and ex vivo. https://www.leica-microsystems.com//science-lab/human-nk-cell-development-requires-cd56-mediated-motility-and-formation-of-the-developmental-synapse/ Wed, 15 Feb 2017 07:36:00 +0000 Ph.D. Emily M. Mace https://www.leica-microsystems.com/18940 EM Sample Preparation Visualization of Membrane Dynamics with Millisecond Temporal Resolution Application Note for Leica EM ICE, Leica EM AFS2 - Electrical stimulation of neurons combined with high-pressure freezing allows physiological activation of synaptic activity and precise control over the time frame of the induced synaptic activity. https://www.leica-microsystems.com//science-lab/visualization-of-membrane-dynamics-with-millisecond-temporal-resolution/ Mon, 07 Nov 2016 10:53:00 +0000 PhD Shigeki Watanabe https://www.leica-microsystems.com/18905 Super-Resolution Neuroscience Botulinum Neurotoxin Type-A Enters a Non-Recycling Pool of Synaptic Vesicles Neuronal communication relies on synaptic vesicles undergoing regulated exocytosis and recycling for multiple rounds of fusion. Whether all synaptic vesicles have identical protein content has been challenged, suggesting that their recycling ability may differ greatly. Botulinum neurotoxin type-A (BoNT/A) is a highly potent neurotoxin that is internalized in synaptic vesicles at motor nerve terminals and induces flaccid paralysis. Recently, BoNT/A was also shown to undergo retrograde transport, suggesting it might enter a specific pool of synaptic vesicles with a retrograde trafficking fate. Using high-resolution microscopy techniques including electron microscopy and single molecule imaging, we found that the BoNT/A binding domain is internalized within a subset of vesicles that only partially co-localize with cholera toxin B-subunit and have markedly reduced VAMP2 immunoreactivity. https://www.leica-microsystems.com//science-lab/botulinum-neurotoxin-type-a-enters-a-non-recycling-pool-of-synaptic-vesicles/ Wed, 19 Oct 2016 11:51:00 +0000 https://www.leica-microsystems.com/18876 Super-Resolution Abstracts of the 6th European Super-Resolution User-Club Meeting The 6th European Super-Resolution User Club Meeting was held in collaboration with Dr. Timo Zimmermann, CRG, and Dr. Pablo Loza-Alvarez, ICFO, Barcelona. According to the founding principle of the club of keeping close to science, both imaging facilities at the CRG and the ICFO opened their doors to the User Club members, allowing them to explore exciting super-resolution and and nanoscopy applications. The meeting agenda covered highly relevant talks around this year’s central theme “Core Facilities and Super-Resolution Microscopy”, as well as plenty of opportunities to network amongst super-resolution users from different European countries. Here we present the abstracts of the talks held during the meeting. https://www.leica-microsystems.com//science-lab/abstracts-of-the-6th-european-super-resolution-user-club-meeting/ Tue, 18 Oct 2016 08:59:00 +0000 Dr. Timo Zimmermann, Dr. Pablo Loza-Alvarez, Dr. Alberto Lleó, Dr. Gražvydas Lukinavicius, Prof. Philip Tinnefeld, Prof. Hans-Georg Kräusslich, Dr. Steffen Dietzel, Dr. Valeria Caiolfa, Lorenzo Albertazzi https://www.leica-microsystems.com/18301 EM Sample Preparation Freeze-Fracture Replication of Pyramidal Cells Application Note for Leica EM HPM100 - Frozen samples (90 μm thick slices frozen by HPM100) were inserted into a double replica table and then fractured into two pieces at –130°C (after insertion of the tissue into BAF 060 the samples should be left in the chamber for 20 min to reach the –130°C). https://www.leica-microsystems.com//science-lab/freeze-fracture-replication-of-pyramidal-cells/ Thu, 08 Sep 2016 16:23:00 +0000 Akos Kulik https://www.leica-microsystems.com/17957 Super-Resolution Confocal Microscopy Light Sheet Microscopy Neuroscience Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence imaging (SeeDB2). SeeDB2 matches the refractive indices of fixed tissues to that of immersion oil (1.518), thus minimizing both light scattering and spherical aberrations. https://www.leica-microsystems.com//science-lab/super-resolution-mapping-of-neuronal-circuitry-with-an-index-optimized-clearing-agent/ Wed, 27 Apr 2016 10:07:00 +0000 https://www.leica-microsystems.com/16593 Super-Resolution Neuroscience Super-Resolution Microscopy of the Synaptic Active Zone At the presynaptic active zone (AZ) a variety of specialized proteins are assembled to complex architectures, which set the basis for speed, precision and plasticity of synaptic transmission. Recently, super-resolution microscopy (SRM) techniques have begun to enter the neurosciences. These approaches combine high spatial resolution with the molecular specificity of fluorescence microscopy. Here, we discuss how SRM techniques can be used to obtain information on the organization of AZ proteins. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-of-the-synaptic-active-zone/ Tue, 15 Dec 2015 10:15:00 +0000 Nadine Ehmann https://www.leica-microsystems.com/14658 Super-Resolution Neuroscience A New Probe for Super-Resolution Imaging of Membranes Elucidates Trafficking Pathways The molecular composition of the organelles involved in membrane recycling is difficult to establish as a result of the absence of suitable labeling tools. We introduce in this paper a novel probe, named membrane-binding fluorophore-cysteine-lysine-palmitoyl group (mCLING), which labels the plasma membrane and is taken up during endocytosis. https://www.leica-microsystems.com//science-lab/a-new-probe-for-super-resolution-imaging-of-membranes-elucidates-trafficking-pathways/ Fri, 17 Oct 2014 12:16:00 +0000 https://www.leica-microsystems.com/13877 Super-Resolution Neuroscience Super-Resolution Microscopy Helped to Create the First 3D Model of a Synapse A research team from Göttingen, led by Prof. Silvio O. Rizzoli, managed to determine the copy numbers and positions of all important building blocks of a synapse for the first time. This allowed them to reconstruct the first scientifically accurate 3D model of a synapse. https://www.leica-microsystems.com//science-lab/super-resolution-microscopy-helped-to-create-the-first-3d-model-of-a-synapse/ Tue, 01 Jul 2014 14:59:00 +0000 https://www.leica-microsystems.com/13373 Super-Resolution Neuroscience Synaptic Vesicle Recycling: Steps and Principles Synaptic vesicle recycling is one of the best‐studied cellular pathways. Many of the proteins involved are known, and their interactions are becoming increasingly clear. However, as for many other pathways, it is still difficult to understand synaptic vesicle recycling as a whole. https://www.leica-microsystems.com//science-lab/synaptic-vesicle-recycling-steps-and-principles/ Mon, 26 May 2014 09:30:00 +0000 Prof. Silvio Rizzoli https://www.leica-microsystems.com/13280 EM Sample Preparation Neuroscience Capturing Cellular Dynamics with Millisecond Temporal Resolution The combination of two powerful techniques: optogenetics and high-pressure freezing now makes it possible to visualize a dynamic cellular activity with temporal resolution of 5 milliseconds. By coupling a flash of light with high-pressure freezing, the process of vesicle recycling at the synapses can now be imaged by electron microscopy. https://www.leica-microsystems.com//science-lab/capturing-cellular-dynamics-with-millisecond-temporal-resolution/ Mon, 12 May 2014 13:16:00 +0000 PhD Shigeki Watanabe, PhD Erik M. Jørgensen https://www.leica-microsystems.com/11033 Neuroscience Stereo Microscopy Live-Cell Imaging Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells Mauthner cells (M-cells) are large reticulospinal neurons located in the hindbrain of teleost fish. They are key neurons involved in a characteristic behavior known as the C-start or escape response that occurs when the organism perceives a threat. The M-cell has been extensively studied in adult goldfish where it has been shown to receive a wide range of excitatory, inhibitory and neuromodulatory signals. We have been examining M-cell activity in embryonic zebrafish in order to study aspects of synaptic development in a vertebrate preparation. In the late 1990s Ali and colleagues developed a preparation for patch clamp recording from M-cells in zebrafish embryos, in which the CNS was largely intact. https://www.leica-microsystems.com//science-lab/patch-clamp-recordings-from-embryonic-zebrafish-mauthner-cells/ Fri, 11 Oct 2013 21:40:00 +0000 https://www.leica-microsystems.com/10135 Super-Resolution Widefield Microscopy Image Restoration and Deconvolution Abstracts of the 3rd European Super-Resolution User-Club Meeting The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and widefield super-resolution users from ten European countries took three days’ out to deepen their knowledge on super-resolution techniques and applications and make use of an opportunity for full exchange of experiences. https://www.leica-microsystems.com//science-lab/abstracts-of-the-3rd-european-super-resolution-user-club-meeting/ Mon, 08 Jul 2013 10:27:00 +0000 Prof. Alberto Diaspro, Ph.D. Illaria Testa, Prof. Silvio Rizzoli, Ph.D. Giuseppe Vicidomini, Prof. Ralf Jacob, Ph.D. Eric Hosy, Prof. Colin Sheppard, Zeno Lavagnino https://www.leica-microsystems.com/7401 EM Sample Preparation Capturing Neurotransmitter Receptors and Ion Channels Neurotransmitter receptors and ion channels in the central nervous system are localized to synaptic and extrasynaptic membrane compartments of pre- and postsynaptic elements of neurons. The impact of the activation of these proteins on synaptic integration and regulation of transmitter release depends on their precise location relative to synapses, as well as on the density and coupling of molecules in microcompartments of the cells. High-resolution qualitative and quantitative visualization of membranebound receptors and ion channels is, therefore, essential for understanding their roles in cell communication. https://www.leica-microsystems.com//science-lab/capturing-neurotransmitter-receptors-and-ion-channels/ Thu, 14 Feb 2013 23:00:00 +0000 Daniel Althof, Akos Kulik https://www.leica-microsystems.com/7735 Super-Resolution Widefield Microscopy Image Restoration and Deconvolution Abstracts of the 2nd European Super-Resolution User-Club Meeting The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. With a mixture of engaging talks by key experts in the field of super-resolution microscopy and stimulating discussion sessions, the meeting proved as popular as last year’s event, attracting a wide range of scientists interested in both confocal and widefield super-resolution and sample preparation techniques. https://www.leica-microsystems.com//science-lab/abstracts-of-the-2nd-european-super-resolution-user-club-meeting/ Thu, 01 Nov 2012 23:00:00 +0000 Prof. Hjalmar Brismar, Prof. Dr. Dr. h.c. Stefan Hell, Prof. Christian Eggeling, Dr. Anna Szymborska, Prof. Alberto Diaspro, Ph.D. Eric Hosy, Ph.D. Giuseppe Vicidomini, Hans van der Voort https://www.leica-microsystems.com/5331 Neuroscience Super-Resolution Sharp Live Images from the Mouse Brain To explore the most intricate structures of the brain in order to decipher how it functions – Stefan Hell’s team of researchers at the Max Planck Institute for Biophysical Chemistry in Göttingen has made a significant step closer to this goal. Using the STED microscopy developed by Hell, the scientists have, for the first time, managed to record detailed live images inside the brain of a living mouse. https://www.leica-microsystems.com//science-lab/sharp-live-images-from-the-mouse-brain/ Sun, 04 Mar 2012 23:00:00 +0000 Dr. Sebastian Berning, Dr. Katrin Willig, Dr. Heinz Steffens, Dr. Payam Dibaj, Prof. Dr. Dr. h.c. Stefan Hell https://www.leica-microsystems.com/10159 Super-Resolution 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. https://www.leica-microsystems.com//science-lab/two-color-sted-microscopy-of-living-synapses-using-a-single-laser-beam-pair/ Sun, 20 Nov 2011 15:22:00 +0000 https://www.leica-microsystems.com/10165 Super-Resolution Neuroscience Live-Cell Imaging 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. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-of-actin-dynamics-in-synapses-deep-inside-living-brain-slices/ Wed, 07 Sep 2011 16:39:00 +0000 https://www.leica-microsystems.com/10866 CLEM Fluorescence Microscopy EM Sample Preparation A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms Electron microscopy (EM) achieves the highest spatial resolution in protein localization, but specific protein EM labeling has lacked generally applicable genetically encoded tags for in situ visualization in cells and tissues. Here we introduce ‘"miniSOG"’ (for mini Singlet Oxygen Generator), a fluorescent flavoprotein engineered from Arabidopsis phototropin 2. https://www.leica-microsystems.com//science-lab/a-genetically-encoded-tag-for-correlated-light-and-electron-microscopy-of-intact-cells-tissues-and-organisms/ Sun, 04 Sep 2011 19:37:00 +0000 https://www.leica-microsystems.com/3000 Live-Cell Imaging Neuroscience Image Restoration and Deconvolution Mapping Billions of Synapses with Microscopy and Mathematics A combination of widefield imaging techniques and image segmentation analysis enable researchers to map learning-induced functional changes in individual synapses throughout the hippocampus. https://www.leica-microsystems.com//science-lab/mapping-billions-of-synapses-with-microscopy-and-mathematics/ Tue, 12 Apr 2011 22:00:00 +0000 Dr. Christopher S. Rex, Allison Paradise https://www.leica-microsystems.com/2727 Neuroscience Super-Resolution 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. https://www.leica-microsystems.com//science-lab/restless-receptors/ Mon, 01 Nov 2010 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, PhD Daniel Choquet https://www.leica-microsystems.com/2471 Neuroscience Super-Resolution 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 https://www.leica-microsystems.com//science-lab/the-missing-link-to-the-nanocosm-of-life/ Mon, 01 Nov 2010 23:00:00 +0000 https://www.leica-microsystems.com/10214 Super-Resolution Neuroscience 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. https://www.leica-microsystems.com//science-lab/the-fate-of-synaptic-vesicle-components-upon-fusion/ Fri, 01 Oct 2010 11:41:00 +0000 Dr. Felipe Opazo https://www.leica-microsystems.com/4138 Super-Resolution Neuroscience 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. https://www.leica-microsystems.com//science-lab/observing-lifes-nanostructures-with-sted/ Mon, 16 Mar 2009 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, Prof. Dr. Stephan Sigrist, Prof. Silvio Rizzoli, Dr. Gregorz Wilczynski