1Cancer and Blood Diseases Institute, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, 2IBM
The present protocol describes a novel method of identifying a population of enucleating orthochromatic erythroblasts by multi-spectral imaging flow cytometry, providing a visualization of the erythroblast enucleation process.
Published June 6, 2014. Keywords: Basic Protocol, Erythropoiesis, Erythroblast enucleation, Reticulocyte, Multi-Spectral Imaging Flow Cytometry, FACS, Multiparameter high-speed cell imaging in flow, Aspect ratio, Delta centroid XY
1Division of Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, 2Center for Regenerative Therapeutics, Brigham and Women's Hospital, 3Harvard Medical School, Harvard University, 4Harvard Stem Cell Institute, Harvard University, 5Harvard-MIT Division of Health Sciences and Technology, 6Department of Mechanical Engineering, Massachusetts Institute of Technology
This study used a multi-well plate microfluidic system, significantly increasing throughput of cell rolling studies under physiologically relevant shear flow. Given the importance of cell rolling in the multi-step cell homing cascade and the importance of cell homing following systemic delivery of exogenous populations of cells in patients, this system offers potential as a screening platform to improve cell-based therapy.
Published October 16, 2013. Keywords: Bioengineering, Microfluidics, Endothelial Cells, Leukocyte Rolling, HL-60 cells, TNF-α, P-selectin, E-selectin
1Department of Psychology, University of Pennsylvania
Informational connectivity measures the correspondence between time courses of multi-voxel information across different brain regions. Multi-voxel pattern discriminability time series are extracted from regions and compared, revealing networks that are not identified in a typical functional connectivity approach.
Published July 1, 2014. Keywords: Neuroscience, fMRI, MVPA, connectivity, informational connectivity, functional connectivity, networks, multi-voxel pattern analysis, decoding, classification, method, multivariate
1Department of Biomedical Engineering, Washington University in St. Louis
We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.
Published January 25, 2013. Keywords: Neuroscience, Neurobiology, Biomedical Engineering, Bioengineering, Physiology, Anatomy, Cellular Biology, Molecular Biology, Entomology, Olfactory Receptor Neurons, Sensory Receptor Cells, Electrophysiology, Olfactory system, extracellular multi-unit recordings, first-order olfactory receptor neurons, second-order projection neurons, third-order Kenyon cells, neurons, sensilla, antenna, locust, Schistocerca Americana, animal model
JoVE Immunology and Infection
1Protein Crystallization Lab, Emerald Bio, 2Molecular Biology Lab, Emerald Bio, 3Scientific Sales Representative, Emerald Bio, 4Group Leader II, Emerald Bio, 5Group Leader I, Emerald Bio, 6Chair of Advisory Board, Emerald Bio, 7Director of Multi-Target Services, Emerald Bio, 8Senior Project Leader, Emerald Bio, 9Project Leader II & SSGCID Site Manager, Emerald Bio
Structure-based drug design plays an important role in drug development. Pursuing multiple targets in parallel greatly increases the chance of success for lead discovery. The following article highlights how the Seattle Structural Genomics Center for Infectious Disease utilizes a multi-target approach for gene-to-structure determination of the PB2 influenza A subunit.
Published June 28, 2013. Keywords: Infection, Structural Biology, Virology, Genetics, Medicine, Biomedical Engineering, Molecular Biology, Infectious Diseases, Microbiology, Genomics, high throughput, multi-targeting, structural genomics, protein crystallization, purification, protein production, X-ray crystallography, Gene Composer, Protein Maker, expression, E. coli, fermentation, influenza, virus, vector, plasmid, cell, cell culture, PCR, sequencing
1Graduate School of Biomedical Science, Neuroscience Program, University of Texas, 2Department of Neurobiology and Anatomy, University of Texas
A fundamental issue in our understanding of cortical circuitry is how networks in different cortical layers encode sensory information. Here we describe electrophysiological techniques utilizing multi-contact laminar electrodes to record single-units and local field potentials and present analyses to identify cortical layers.
Published September 8, 2011. Keywords: Neuroscience, laminar probes, cortical layers, local-field potentials, population coding
1Laboratory of Neural Microcircuitry - Brain Mind Institute, Ecole Polytechnique Federale de Lausanne
Multi-electrode patch-clamp recordings constitute a complex task. Here we show how, by automating of many of the experimental steps, it is possible to accelerate the process leading to qualitative improvement in performance and number of recordings.
Published October 18, 2013. Keywords: Neuroscience, Patch-clamp, automatic positioning, whole-cell, neuronal recording, in vitro, multi-electrode
1Center for Systems Biology, Massachusetts General Hospital
We present a method for rapid, reversible immobilization of small molecules and functionalized nanoparticle assemblies for Surface Plasmon Resonance (SPR) studies, using sequential on-chip bioorthogonal cycloaddition chemistry and antibody-antigen capture.
Published September 23, 2013. Keywords: Chemistry, Organic Chemicals, Macromolecular Substances, Chemistry and Materials (General), Surface Plasmon Resonance, Bioorthogonal Chemistry, Diels-Alder Cycloaddition Reaction, Small Molecule Immobilization, Binding Kinetics, Immobilized Nanoparticles
1Department of Biology, University of Washington
Olfactory cues mediate many different behaviors in insects, and are often complex mixtures comprised of tens to hundreds of volatile compounds. Using gas chromatography with multi-channel recording in the insect antennal lobe, we describe a method for the identification of bioactive compounds.
Published February 24, 2013. Keywords: Neuroscience, Neurobiology, Physiology, Biochemistry, Chemistry, Entomlogy, Behavior, electrophysiology, olfaction, olfactory system, insect, multi-channel recording, gas chromatography, pollination, bees, Bombus impatiens, antennae, brain, animal model
1Department of Anesthesiology & Pain Medicine, Mitochondria and Metabolism Center, University of Washington, Seattle
A spectrofluorometric protocol for the measurement of the mitochondrial permeability transition pore opening in isolated mouse heart mitochondria is presented here. The assay involves the simultaneous measurement of mitochondria Ca2+ handling, mitochondrial membrane potential and mitochondrial volume. The procedure for obtaining high-quality and functional heart mitochondria is also described.
Published September 7, 2012. Keywords: Cellular Biology, Mitochondria, respiration, mitochondrial permeability transition pore (mPTP), membrane potential, swelling, calcium, spectrofluorometer