Leica Science Lab - Tag : Neuron https://www.leica-microsystems.com//science-lab/tag/tags/neuron/show/Tag/ Article tagged with Neuron 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/20202 Super-Resolution Researchers Find a “Digital” Mechanism Behind Neuronal Changes from Learning Neurons react to learning and memory by activating synaptic connections. The mechanisms behind this fundamental process are complex and poorly understood. Researchers at Thomas Jefferson University have found that neuron plasticity operates in a “digital” fashion through nanomodules of discrete size that multiply and strengthen neuronal connections upon stimulation. This breakthrough was published on April 23rd in the journal Nature Neuroscience. https://www.leica-microsystems.com//science-lab/researchers-find-a-digital-mechanism-behind-neuronal-changes-from-learning/ Mon, 04 Jun 2018 22:00:00 +0000 Dr. Martin Hruska, Dr. Julia Roberti 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/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/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/18146 EM Sample Preparation Cultured Rat Hippocampal Neurons Application Note for Leica EM ICE - Rat Hippocampal neurons, cultured on 50 μm thick Aclar (Aclar embedding film, EMS) for 19 days, were frozen in the 100 μm deep side of lecithin coated (detailed protocol Appendix I) type A 3 mm Cu/Au carriers (Leica) and sandwiched with the flat side of lecithin coated type B 3 mm Cu/Au carriers (Leica). No additional filler was used, only cell culture medium with the addition of Hepes buffer pH 7.2 to a final concentration of 25 mM. Samples were frozen in a high-pressure freezer (Leica EM ICE). https://www.leica-microsystems.com//science-lab/cultured-rat-hippocampal-neurons/ Mon, 29 Aug 2016 07:12:00 +0000 E. G. van Donselaar, Dr. Martin Harterink, Drs. C. E. M. Vocking, Dr. W. H. Mueller, Prof. Dr. C. C. Hoogenraad https://www.leica-microsystems.com/15645 Confocal Microscopy Live-Cell Imaging Label-free in vivo Imaging of Myelinated Axons in Health and Disease with Spectral Confocal Reflectance Microscopy We report a new technique for high-resolution in vivo imaging of myelinated axons in the brain, spinal cord and peripheral nerve that requires no fluorescent labeling. This method, based on spectral confocal reflectance microscopy (SCoRe), uses a conventional laser scanning confocal system to generate images by merging the simultaneously reflected signals from multiple lasers of different wavelengths. https://www.leica-microsystems.com//science-lab/label-free-in-vivo-imaging-of-myelinated-axons-in-health-and-disease-with-spectral-confocal-reflectance-microscopy/ Fri, 01 Jul 2016 09:29:00 +0000 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/15762 Confocal Microscopy Neuroscience Switching Roles: The Functional Plasticity of Adult Tissue Stem Cells Adult organisms have to adapt to survive, and the same is true for their tissues. Rates and types of cell production must be rapidly and reversibly adjusted to meet tissue demands in response to both local and systemic challenges. Recent work reveals how stem cell (SC) populations meet these requirements by switching between functional states tuned to homoeostasis or regeneration. https://www.leica-microsystems.com//science-lab/switching-roles-the-functional-plasticity-of-adult-tissue-stem-cells/ Mon, 18 May 2015 12:56:00 +0000 https://www.leica-microsystems.com/14367 Neuroscience Live-Cell Imaging Fluorescence Microscopy Confocal Microscopy Axon Injury and Regeneration in the Adult Drosophila Neural regeneration is a fascinating process with profound impact on human health, such that defining biological and genetic pathways is of interest. Here we describe an in vivo preparation for neuronal regeneration in the adult Drosophila. The nerve along the anterior margin of the wing is comprised of ~225 neurons that send projections into the central neuropil (thorax). https://www.leica-microsystems.com//science-lab/axon-injury-and-regeneration-in-the-adult-drosophila/ Fri, 02 Jan 2015 21:01:00 +0000 https://www.leica-microsystems.com/13948 Neuroscience Video Talk by Karl Deisseroth: Optogenetics Optogenetics is a combination of genetics and optics to achieve a gain or loss of function of biochemical events such as action potentials in a particular neuron or tissue. Opsin genes encode proteins that receive light and give rise to ion flow. This talk gives an introduction to optogenetics followed by examples of how optogenetics is being used to study the brain. https://www.leica-microsystems.com//science-lab/video-talk-by-karl-deisseroth-optogenetics/ Tue, 18 Nov 2014 12:52:00 +0000 https://www.leica-microsystems.com/14246 Super-Resolution Neuroscience Correlated Optical and Isotopic Nanoscopy The isotopic composition of different materials can be imaged by secondary ion mass spectrometry. In biology, this method is mainly used to study cellular metabolism and turnover, by pulsing the cells with marker molecules such as amino acids labelled with stable isotopes (15N, 13C). The incorporation of the markers is then imaged with a lateral resolution that can surpass 100 nm. https://www.leica-microsystems.com//science-lab/correlated-optical-and-isotopic-nanoscopy/ Fri, 22 Aug 2014 09:53:00 +0000 https://www.leica-microsystems.com/13740 Super-Resolution Neuroscience Interview with Dr. Yasushi Okada Yasushi Okada, team leader at Riken Quantitative Biology Center in Osaka, Japan, investigates vesicular transport mechanisms in neuronal cells. As the size of transported vesicles is below 100 nm and the diameter of microtubules is about 25 nm, he uses super-resolution techniques to study the sophisticated machinery of neuronal transport. https://www.leica-microsystems.com//science-lab/interview-with-dr-yasushi-okada/ Tue, 03 Jun 2014 15:38:00 +0000 Dipl. oec.-troph. Anja Schué, MD, PhD Yasushi Okada, PhD Isabelle Köster 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/11000 Confocal Microscopy Neuroscience Live-Cell Imaging Imaging Pheromone Sensing in a Mouse Vomeronasal Acute Tissue Slice Preparation In mice, the ability to detect pheromones is principally mediated by the vomeronasal organ (VNO). Here, an acute tissue slice preparation of VNO for performing calcium imaging is described. This physiological approach allows observations of subpopulations and/or individual neurons in a living tissue and is convenient for receptor-ligand identification. https://www.leica-microsystems.com//science-lab/imaging-pheromone-sensing-in-a-mouse-vomeronasal-acute-tissue-slice-preparation/ Fri, 02 May 2014 14:24:00 +0000 https://www.leica-microsystems.com/2469 Laser Microdissection Neuroscience The Morbus Parkinson Puzzle A characteristic sign of M. Parkinson is the deterioration of dopaminergic neurons in the mid-brain, specifically in the substantia nigra (SN, black substance). Different causes and forms of this disease have been identified. In the case of the genetic familial form, for example, it has been possible to identify various genes that have a causal influence for M. Parkinson. https://www.leica-microsystems.com//science-lab/the-morbus-parkinson-puzzle/ Thu, 27 Mar 2014 11:11:00 +0000 Dr. Falk Schlaudraff, Dr. Olaf Spörkel https://www.leica-microsystems.com/11035 Stereo Microscopy Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors while the second involves a wholesale (global) activation of most or all such neurons. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons. https://www.leica-microsystems.com//science-lab/local-and-global-methods-of-assessing-thermal-nociception-in-drosophila-larvae/ Fri, 14 Feb 2014 12:46:00 +0000 https://www.leica-microsystems.com/12449 Laser Microdissection Neuroscience Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection High-quality total RNA has been prepared from cell bodies of mouse spinal cord motor neurons by laser capture microdissection after staining spinal cord sections with Azure B in 70% ethanol. Sufficient RNA (~40–60 ng) is recovered from 3,000–4,000 motor neurons to allow downstream RNA analysis by RNA-seq and qRT-PCR. https://www.leica-microsystems.com//science-lab/production-of-rna-for-transcriptomic-analysis-from-mouse-spinal-cord-motor-neuron-cell-bodies-by-laser-capture-microdissection/ Thu, 30 Jan 2014 09:11:00 +0000 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/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/6773 Laser Microdissection K-ATP Channels in Dopamine Substantia Nigra Neurons Control Bursting and Novelty-induced Exploration Phasic activation of the dopamine (DA) midbrain system in response to unexpected reward or novelty is critical for adaptive behavioral strategies. This activation of DA midbrain neurons occurs via a synaptically triggered switch from low-frequency background spiking to transient high-frequency burst firing. We found that, in medial DA neurons of the substantia nigra (SN), activity of ATP-sensitive potassium (K-ATP) channels enabled NMDA-mediated bursting in vitro as well as spontaneous in vivo burst firing in anesthetized mice. https://www.leica-microsystems.com//science-lab/k-atp-channels-in-dopamine-substantia-nigra-neurons-control-bursting-and-novelty-induced-exploration/ Thu, 04 Oct 2012 22:00:00 +0000 Julia Schiemann, Dr. Falk Schlaudraff, Dr. Verena Klose, Markus Bingmer, Prof. Susumu Seino, Dr. Peter J. Magill, Ph.D. Kareem A. Zaghloul, Dr. Gaby Schneider, Prof. Birgit Liss, Prof. Jochen Röper 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/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/2470 Live-Cell Imaging Neuroscience Sniffing Out the Secrets of Social Behavior Yet we are only just beginning to understand the complexities and functional differences of the sense of smell in mammals. Prof. Marc Spehr, head of the Department of Chemosensation at RWTH Aachen University since 2009, explains his findings on the neuronal mechanisms of olfactory perception and signal processing using the mouse model. He and his team are trying to find out how substances for social interaction are perceived and how this perception generates a specific type of behavior. https://www.leica-microsystems.com//science-lab/sniffing-out-the-secrets-of-social-behavior/ Mon, 01 Nov 2010 23:00:00 +0000 Dipl. oec.-troph. Anja Schué, Prof. Dr. Marc Spehr https://www.leica-microsystems.com/2732 Confocal Microscopy Multiphoton Microscopy Live-Cell Imaging Neuroscience Exploring the Concert of Neuronal Activities Brain research using Confocal and Multiphoton Microscopy. Using imaging techniques such as confocal and two-photon microscopy, neuronal dendritic arborization of neurons and their synaptic interconnections can be visualized. https://www.leica-microsystems.com//science-lab/exploring-the-concert-of-neuronal-activities/ Mon, 01 Nov 2010 23:00:00 +0000 Ph.D. Randy Bruno, Ph.D. Myriam Gastard https://www.leica-microsystems.com/9687 Neuroscience Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells We will demonstrate how to study the functional effects of introducing a point mutation in an ion channel. We study G protein-gated inwardly rectifying potassium (referred to as GIRK) channels, which are important for regulating the excitability of neurons. There are four different mammalian GIRK channel subunits (GIRK1-GIRK4) – we focus on GIRK2 because it forms a homotetramer. https://www.leica-microsystems.com//science-lab/mutagenesis-and-functional-analysis-of-ion-channels-heterologously-expressed-in-mammalian-cells/ Fri, 01 Oct 2010 16:35:00 +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/8111 Neuroscience Live-Cell Imaging New Standard in Electrophysiology and Deep Tissue Imaging The function of nerve and muscle cells relies on ionic currents flowing through ion channels. These ion channels play a major role in cell physiology. One way to investigate ion channels is to use patch clamping. This method allows investigation of ion channels in detail and recording of the electric activity of different types of cells, mainly excitable cells like neurons, muscle fibres or beta cells of the pancreas. The patch clamping technique was developed by Erwin Neher and Bert Sakmann in the 1970s and 80s to study individual ion channels in living cells. In 1991 they received the Nobel Prize for Physiology and Medicine for their work. Today the patch clamping technique is one of the most important methods in the field of electrophysiology. https://www.leica-microsystems.com//science-lab/new-standard-in-electrophysiology-and-deep-tissue-imaging/ Tue, 17 Mar 2009 22:04:00 +0000 Dr. Irmtraud Steinmetz 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