JoVE Applied Physics
1Physics and Astronomy Department, Vassar College, 2Biology Department, Vassar College
The technique presented here measures the path of freely swimming microscopic species using single wavelength exposure. C. elegans are used to demonstrate shadow imaging as an inexpensive alternative to costly microscopes. This technique can be adapted to accommodate various orientations, environments and species to measure direction, speed, acceleration and forces.
Published April 18, 2014. Keywords: Physics, C. elegans, nematode, shadow imaging, locomotion, video analysis, swimming behavior, force
1Synthetic Biology Platform, Wyss Institute for Biologically Inspired Engineering, Harvard University, 2Department of Systems Biology, Harvard Medical School, 3Department of Biotechnology, Delft University of Technology
This work describes a novel method for selectively targeting subcellular organelles in plants, assayed using the BioRad Gene Gun.
Published April 18, 2014. Keywords: Environmental Sciences, Plant Leaves, Synthetic Biology, Plants, Genetically Modified, DNA, Plant, RNA, Gene Targeting, Plant Physiological Processes, Genes, Gene gun, Gibson assembly, Nicotiana benthamiana, Alternative splicing, confocal microscopy, chloroplast, peroxisome
JoVE Applied Physics
1Department of Bioengineering and Therapeutic Sciences, Unversity of California, San Francisco
We have developed a technique for picoinjecting microfluidic drops that does not require metal electrodes. As such, devices incorporating our technique are simpler to fabricate and to use.
Published April 18, 2014. Keywords: Bioengineering, Droplet microfluidics, picoinjection, lab on a chip, electrodes, microfabrication
1INSERM UMR-910, Aix-Marseille University, 2Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego
We describe a series of methods to inject dyes, DNA vectors, virus, and cells in order to monitor both cell fate and phenotype of endogenous and grafted cells derived from embryonic or pluripotent cells within mouse embryos at embryonic day (E)9.5 and later stages of development.
Published April 17, 2014. Keywords: Developmental Biology, Cell, DNA, dye injection, mouse embryo, embryo culture, ultrasound, mouse heart, stem cells
1Department of Biomedical Engineering, Carnegie Mellon University, 2Department of Materials Science and Engineering, Carnegie Mellon University
A method to obtain nanofibers and complex nanostructures from single or multiple extracellular matrix proteins is described. This method uses protein-surface interactions to create free-standing protein-based materials with tunable composition and architecture for use in a variety of tissue engineering and biotechnology applications.
Published April 17, 2014. Keywords: Bioengineering, Nanofibers, Nanofabrics, Extracellular Matrix Proteins, Microcontact Printing, Fibronectin, Laminin, Tissue Engineering, poly(N-isopropylacrylamide), Surface-Initiated Assembly
1Department of Cell Biology and Physiology, Washington University School of Medicine
Four-dimensional (4D) imaging is utilized to study the behavior and interactions among two types of endosomes in living vertebrate nerve terminals. Movement of these small structures is characterized in three dimensions, permitting confirmation of events such as endosome fusion and exocytosis.
Published April 16, 2014. Keywords: Neuroscience, Microscopy, Fluorescence, Endocytosis, nerve, endosome, lysosome, deconvolution, 3D, 4D, epifluorescence
1Graduate Program in Neuroscience, Rutgers University, 2Department of Biomedical Engineering, Rutgers University, 3Center for Cognitive Science, Rutgers University, 4Department of Psychology, Rutgers University, 5Department of Computer Science, Rutgers University
It is unclear how top-down signals from the ventral visual stream affect movement. We developed a paradigm to test motor behavior towards a target on a 3D depth inversion illusion. Significant differences are reported in both deliberate, goal-directed movements and automatic actions under illusory and veridical viewing conditions.
Published April 16, 2014. Keywords: Behavior, vision for action, vision for perception, motor control, reach, grasp, visuomotor, ventral stream, dorsal stream, illusion, space perception, depth inversion
1Department of Veterans Affairs, Minneapolis Veterans Affairs Health Care System, 2Department of Food Science and Nutrition, University of Minnesota, 3Department of Integrative Biology and Physiology, University of Minnesota, 4Department of Medicine, University of Minnesota Medical School, University of Minnesota
Multiplex assays can provide beneficial information for basic cellular mechanisms and eliminate waste of reagents and unnecessary repetitive experiments. We describe here a multiplex caspase-3/7 activity assay, using fluorescent- and luminescent-based methods, to determine cell viability in an in vitro hypothalamic model following oxidative challenge with palmitic acid.
Published April 16, 2014. Keywords: Neuroscience, apoptosis, obesity, caspase, resazurin, DEVD, palmitic acid, hypothalamic model
JoVE Immunology and Infection
1Department of Neurology, University of Münster, 2Interdisciplinary Center for Clinical Research (IZKF), Münster, 3Institute of Physiology – Neuropathophysiology, University of Münster
Experimental autoimmune encephalomyelitis (EAE) is an established animal model of multiple sclerosis. C57BL/6 mice are immunized with myelin oligodendrocyte glycoprotein (MOG) peptide 35-55 (MOG35-55), resulting in an ascending flaccid paralysis caused by autoreactive immune cells in the central nervous system. Protocols for disease induction and monitoring will be discussed.
Published April 15, 2014. Keywords: Immunology, experimental autoimmune encephalomyelitis, EAE, multiple sclerosis, MS, animal model, Autoimmunity, neuroinflammation, central nervous system, pertussis
JoVE Immunology and Infection
1Department of Clinical Neurosciences, John Van Geest Centre for Brain Repair, University of Cambridge, UK, 2NIHR Biomedical Research Center, University of Cambridge, UK
The transplantation of neural stem/progenitor cells (NPCs) holds great promises in regenerative neurology. The systemic delivery of NPCs has turned into effective, low invasive, and therapeutically very efficacious protocol to deliver stem cells in the brain and spinal cord of rodents and nonhuman primates affected by experimental chronic inflammatory damage of the central nervous system.
Published April 15, 2014. Keywords: Immunology, Somatic neural stem/precursor cells, neurodegenerative disorders, regenerative medicine, multiple sclerosis, experimental autoimmune encephalomyelitis, systemic delivery, intravenous, intracerebroventricular