JoVE Immunology and Infection
1Department of Immunology, The University of Texas MD Anderson Cancer Center, 2The University of Texas Graduate School of Biomedical Sciences
This protocol describes experimental procedures to assess the differentiation of plasmacytoid dendritic cells in Peyer’s patch from common dendritic cell progenitors, using techniques involving FACS-mediated cell isolation, hydrodynamic gene transfer, and flow analysis of immune subsets in Peyer’s patch.
Published March 17, 2014. Keywords: Immunology, hematopoiesis, dendritic cells, Peyer's patch, cytokines, adoptive transfer
1Anatomical Institute, Department of Biomedicine, University of Basel
We present a protocol that permits to view and to quantitatively asses the morphology of the dendritic tree of individual Purkinje cells grown in organotypic cerebellar slice cultures. This protocol is intended to promote studies on the mechanisms of Purkinje cell dendritic development.
Published March 21, 2012. Keywords: Neuroscience, dendritic development, dendritic branching, cerebellum, Purkinje cells
1Department of Physiology, Northwestern University Feinberg School of Medicine, 2Department of Psychiatry and Behavioral Sciences, Northwestern University Feinberg School of Medicine
Numerous recent studies have identified mutations in synaptic proteins associated with brain pathologies. Primary cultured cortical neurons offer great flexibility in examining the effects of these disease-associated proteins on dendritic spine morphology and motility.
Published July 13, 2011. Keywords: Neuroscience, Excitatory synapse, neuroscience, brain, cortex, cortical neurons, primary culture, confocal microscopy, time-lapse imaging, remodeling.
1Center for Neuroscience, Swammerdam Institute for Life Sciences, University of Amsterdam, 2Van Leeuwenhoek Centre for Advanced Microscopy, Section Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam
This article describes a working protocol to image dendritic spines from hippocampal neurons in vitro using Structured Illumination Microscopy (SIM). Super-resolution microscopy using SIM provides image resolution significantly beyond the light diffraction limit in all three spatial dimensions, allowing the imaging of individual dendritic spines with improved detail.
Published May 4, 2014. Keywords: Neuroscience, Dendritic Spine, Microscopy, Confocal, Fluorescence, Neurosciences, hippocampus, primary neuron, super resolution microscopy, structured illumination microscopy (SIM), neuroscience, dendrite
1Department of Molecular Biosciences, University of California, Davis
We describe a protocol for using bone morphogenetic protein-7 (BMP-7) or Matrigel to selectively induce dendritic growth in primary sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rats.
Published March 21, 2012. Keywords: Neuroscience, Bone morphogenetic proteins (BMPs), Matrigel, dendrite, dendritogenesis, neuronal morphogenesis, sympathetic neurons
JoVE Immunology and Infection
1Department of General Neurology, Hertie Institute for Clinical Brain Research, 2Institute of Pharmacology, University of Bern, 3Department of Immunology, University Medical Center Hamburg-Eppendorf, 4Department of Thoracic and Cardiovascular Surgery, University Clinic Tuebingen, 5Department of Neurology, University Hospital Erlangen
This protocol details a method to isolate antigen presenting cells from human thymus via different steps of enzymatic digestion of the tissue followed by density centrifugation of the single cell suspension and finally magnetic and/or FACS sorting of the cell populations of interest.
Published September 19, 2013. Keywords: Immunology, Immune System Processes, Biological Processes, immunology, Immune System Diseases, Immune System Phenomena, Life Sciences (General), immunology, human thymus, isolation, dendritic cells, mTEC, cTEC
1Department of Cellular and Molecular Physiology, Yale University School of Medicine
An imaging technique for monitoring of membrane potential changes with sub-micrometer spatial and sub-millisecond temporal resolution is described. The technique, based on laser excitation of voltage-sensitive dyes, allows measurements of signals in axons and axon collaterals, terminal dendritic branches, and individual dendritic spines.
Published November 29, 2012. Keywords: Neuroscience, Medicine, Physiology, Molecular Biology, Cellular Biology, voltage-sensitive dyes, brain, imaging, dendritic spines, axons, dendrites, neurons
1Department of Molecular Cell and Developmental Biology, University of California, Santa Cruz
Time-lapse imaging in the living animal provides valuable information on structural reorganization in the intact brain. Here, we introduce a thinned-skull preparation that allows transcranial imaging of fluorescently labeled synaptic structures in the living mouse cortex by two-photon microscopy.
Published May 12, 2014. Keywords: Neuroscience, dendritic spine, mouse cortex, in vivo, two-photon microscopy, thinned-skull, imaging
JoVE Immunology and Infection
1Experimental and Clinical Research Center, A joint cooperation between the Charité Medical Faculty and the Max Delbrück Center for Molecular Medicine, 2Berlin Ultrahigh Field Facility (B.U.F.F.), Max Delbrück Center for Molecular Medicine
Tracking of cells using MRI has gained remarkable attention in the past years. This protocol describes the labeling of dendritic cells with fluorine (19F)-rich particles, the in vivo application of these cells, and monitoring the extent of their migration to the draining lymph node with 19F/1H MRI and 19F MRS.
Published March 20, 2013. Keywords: Molecular Biology, Immunology, Cellular Biology, Physiology, Anatomy, Biomedical Engineering, Hematology, nuclear magnetic resonance, NMR, Fluorine, dendritic cells, migration, lymph nodes, magnetic resonance imaging, MRI, magnetic resonance spectroscopy, MRS, spectroscopy, imaging, cell tracking, clinical techniques
1Canadian Centre for Behavioural Neuroscience, University of Lethbridge
This paper describes the procedures for tactile stimulation of rat pups and subsequent Golgi-Cox staining of neuronal morphology. Tactile stimulation is a positive experience that is administered in the perinatal period by stroking pups with a household duster. Golgi-cox staining is a reliable procedure permitting the visualization of entire neurons.
Published September 25, 2013. Keywords: Neuroscience, Brain, Prefrontal Cortex, Neurons, Massage, Staining and Labeling, mPFC, spine density, methodology, enrichment