JoVE Immunology and Infection
1Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Harvard Medical School, 2Department of Mechanical and Aerospace Engineering, The Ohio State University, 3Center for Computational and Integrative Biology, Massachusetts General Hospital, Harvard Medical School, 4Dept. of Chemical and Biomolecular Engineering, Vanderbilt University
A method is described to individually select, manipulate, and image live pathogens using an optical trap coupled to a spinning disk microscope. The optical trap provides spatial and temporal control of organisms and places them adjacent to host cells. Fluorescence microscopy captures dynamic intercellular interactions with minimal perturbation to cells.
Published July 28, 2011. Keywords: Immunology, Optical trapping, optical tweezers, T-cell, pathogen, live cell imaging, spinning disk confocal microscopy, Aspergillus fumigatus, Candida albicans, fungi
1Department of Biology, Pace University, 2Cellular and Molecular Medicine, University of California, San Diego, 3Division of Cell Biology and Cell Physiology, Zoological Institute, Braunschweig University of Technology
The temporal and spatial resolution of genetic manipulations determines the spectrum of biological phenomena that they can perturb. Here we use temporally and spatially discrete in vivo electroporation, combined with transgenic lines of zebrafish, to induce expression of a GFP transgene specifically in neurons of the developing olfactory bulb.
Published August 15, 2011. Keywords: Neuroscience, electroporation, zebrafish, olfactory bulb, Gal4 enhancer trap
JoVE Applied Physics
1Department of Physics and Astronomy, University of Sheffield, 2Department of Chemical and Biological Engineering, The University of Sheffield, 3ISIS Pulsed Neutron and Muon Source Science and Technology Facilities Council, Rutherford Appleton Laboratory
Progress has been made in utilizing spin echo resolved grazing incidence scattering (SERGIS) as a neutron scattering technique to probe the length-scales in irregular samples. Crystallites of [6,6]-phenyl-C61-butyric acid methyl ester have been probed using the SERGIS technique and the results confirmed by optical and atomic force microscopy.
Published January 15, 2014. Keywords: Physics, Spin Echo Resolved Grazing Incidence Scattering, Neutron, Crystallite, Organic solar cell, PCBM, P3HT
1Department of Molecular Biology, Lewis-Sigler Institute for Integrative Genomics, Princeton University, 2Department of Physics, Lewis-Sigler Institute for Integrative Genomics, Princeton University
We present a protocol for bending filamentous bacterial cells attached to a cover-slip surface with an optical trap to measure the cellular bending stiffness.
Published April 26, 2010. Keywords: Microbiology, optical trap, cell mechanics, E. coli, cell bending
JoVE Clinical and Translational Medicine
1Medicine, University of California San Diego - UCSD, 2Bioengineering, University of California San Diego - UCSD, 3Radiology, University of California San Diego - UCSD
A MR imaging method to study the distribution of pulmonary blood flow under a variety of physiological conditions, in this case exposure to three different inspired oxygen concentrations: hypoxia, normoxia, and hyperoxia, is described. This technique utilizes human pulmonary physiology research techniques in an MR scanning environment.
Published May 30, 2011. Keywords: Medicine, arterial spin labeling, lung proton density, functional lung imaging, hypoxic pulmonary vasoconstriction, oxygen consumption, ventilation, magnetic resonance imaging
1Schulich Faculty of Chemistry, The Technion, Israel Institute of Technology
This protocol describes a method for micron-scale three-dimensional imaging of oxygen concentration in the immediate environment of live cells by electron spin resonance microscopy.
Published August 26, 2010. Keywords: Cellular Biology, ESR, EPR, Oxygen, Imaging, microscopy, live cells
1Department of Zoology, University of Oklahoma - Norman, 2Department of Biology, Brandeis University
We describe a Flippase-induced intersectional Gal80/Gal4 repression (FINGR) method, allowing tissue-specific FLP to determine Gal80 expression patterns. Wherever Gal4 and FLP overlap, Gal4 expression is turned on (Gal80 flipped out) or off (Gal80 flipped in). The FINGR method is versatile for clonal analysis and neural circuit mapping.
Published June 3, 2011. Keywords: Neuroscience, UAS, Gal4, Gal80, Flippase, FRT, Clonal analysis, Behavior, Drosophila
1Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, 2Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign
In this article, we present a microfluidic-based method for particle confinement based on hydrodynamic flow. We demonstrate stable particle trapping at a fluid stagnation point using a feedback control mechanism, thereby enabling confinement and micromanipulation of arbitrary particles in an integrated microdevice.
Published January 21, 2011. Keywords: Bioengineering, hydrodynamic, trap, trapping, confinement, micromanipulation, microfluidics, stagnation point flow
1The Davis Heart and Lung Research Institute, The Ohio State University, 2Department of Pharmacology, College of Medicine, The Ohio State University
Electron paramagnetic resonance (EPR) spectroscopy was employed to detect nitric oxide from bovine aortic endothelial cells and superoxide radical anion from human neutrophils using iron (II)-N-methyl-D-glucamine dithiocarbamate, Fe(MGD)2 and 5,5-dimethyl-1-pyroroline-N-oxide, DMPO, respectively.
Published August 18, 2012. Keywords: Molecular Biology, Cellular Biology, Physics, Biophysics, spin trap, eNOS, ROS, superoxide, NO, EPR
1Medical Service, VA Palo Alto Health Care System , Stanford University School of Medicine
The associated chromosome trap (ACT) assay is a novel unbiased method for identifying long-range DNA interactions. The characterization of long range DNA interactions will allow us to determine the relationship of nuclear architecture to gene expression in both normal physiology and in diseased states.
Published April 23, 2011. Keywords: Molecular Biology, Associated chromosomal Trap, DNA long-range interaction, nuclear architecture, gene regulation