JoVE Immunology and Infection
1Department of Biological Sciences, University of Wisconsin Milwaukee
Bacterial pyomelanin production results in increased resistance to oxidative stress and virulence. We report on techniques that can be used to determine inhibition of pyomelanin production and assay the resulting increase in sensitivity to oxidative stress in bacteria, as well as determine antibiotic minimum inhibitory concentration (MIC).
Published August 31, 2015. Keywords: Immunology, Microbiology, Pseudomonas aeruginosa, pyomelanin, NTBC, oxidative stress, minimum inhibitory concentration
1Bloomberg School of Public Health, Environmental Health Sciences, Johns Hopkins University, 2Department of Medicine, Pulmonary Diseases and Critical Care Medicine, University of North Carolina School of Medicine
The goal of this paper is to describe simple methods that will greatly aid in the setup and analysis of mouse lungs with lung cancer or other pathologies. We present 3 protocols to simply and reliably carry out lung instillations, fixation, and lung volume measurements.
Published August 31, 2015. Keywords: Medicine, Lung histology, Lung morphometry, Lung fixation, Lung stereology, Lung volume, Lung Intubation
1Department of Translational Medical Science, Graduate School of Medicine, Mie University, 2Department of Molecular and Cellular Pharmacology, Pharmacogenomics, and Pharmacoinformatics, Graduate School of Medicine, Mie University
Repeated blood sampling is necessary and important in animal research. We developed a novel, non-lethal and reliable method for repeated blood collection from adult zebrafish, and applied this method to the study of blood biochemistry, including glucose metabolism.
Published August 30, 2015. Keywords: Basic Protocol, Repeated blood collection, adult zebrafish, dorsal aorta, hemoglobin, fasting blood glucose, plasma triacylglycerol, total cholesterol, animal model
1Children's Hospital of Eastern Ontario Research Institute, 2Sunnybrook Research Institute, University of Toronto, 3Department of Biochemistry, University of Toronto
Protein phosphorylation is a central feature of how cells interpret and respond to information in their extracellular milieu. Here, we present a high throughput screening protocol using kinases purified from mammalian cells to rapidly identify kinases that phosphorylate a substrate(s) of interest.
Published August 29, 2015. Keywords: Molecular Biology, Molecular Biology, protein phosphorylation, protein kinase, screening, signal transduction, kinomics
1Department of Biomedical Engineering, Boston University, 2Departments of Biomedical Engineering, Chemistry, and Medicine, Boston University
Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.
Published August 28, 2015. Keywords: Bioengineering, Electrospinning, electrospraying, polycaprolactone, poly(lactide-co-glycolide), microfiber, nanofiber, microparticles, superhydrophobic, biomaterials, drug delivery, biodegradable, surface coatings.
1School of Pharmacy, Pacific University, 2Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University
The goal of this protocol is to describe the preparation and characterization of physically entrapped, poorly water soluble drugs in micellar drug delivery systems composed of amphiphilic block copolymers.
Published August 28, 2015. Keywords: Chemistry, Amphiphilic block copolymers, Polymeric micelles, drug delivery, in vitro characterization, chemotherapy, multi-drug loading, nanocarriers
1Division of Cardiovascular Medicine, Department of Medicine, University of Massachusetts Medical School
De novo lipogenesis and β-fatty acid oxidation constitute key metabolic pathways in hepatocyte, pathways that are perturbed in several metabolic disorders, including fatty liver disease. Here we demonstrate isolation of mouse primary hepatocytes and describe quantification of β-fatty acid oxidation and lipogenesis.
Published August 27, 2015. Keywords: Molecular Biology, Liver, Hepatocyte, Mouse, Fatty Acid, Oxidation, Lipogenesis, Metabolism, Palmitate
1Department of Cell & Developmental Biology, SUNY Upstate Medical University
Many mammalian cells preferentially migrate towards a more rigid matrix or substrate through durotaxis. The goal of this protocol is to provide a simple in vitro system that can be used to study and manipulate cell durotaxis behaviors by incorporating polydimethylsiloxane (PDMS) substrates of defined rigidity, interfacing with glass coverslips.
Published August 27, 2015. Keywords: Molecular Biology, cell migration, mechanosensing, durotaxis, polydimethylsiloxane, tumor invasion, extracellular matrix rigidity, mechanotransduction, U2OS
1Department of Materials Science and Engineering, University of California Berkeley, 2Materials Science Division, Lawrence Berkeley National Laboratory, 3Advanced Light Source Division, Lawrence Berkeley National Laboratory, 4Department of Chemical and Biomolecular Engineering, University of California Berkeley, 5Environmental Energy Technology Division, Lawrence Berkeley National Laboratory
Synchrotron-based hard X-ray microtomography is used to image the electrochemical growth of dendrites from a lithium metal electrode through a solid polymer electrolyte membrane.
Published August 26, 2015. Keywords: Engineering, Lithium-ion batteries, lithium dendrite growth, polymer electrolytes, X-ray microtomography, electrochemistry, X-ray imaging
1Electrical Engineering, Stanford University, 2Applied Physics, Stanford University
The scalability and resolution of conventional optical manipulation techniques are limited by diffraction. We circumvent the diffraction limit and describe a method of optically transporting nanoparticles across a chip using a gold surface patterned with a path of closely spaced C-shaped plasmonic resonators.
Published August 26, 2015. Keywords: Engineering, Plasmonics, optical trapping, near-field trapping, nano-optical conveyor belt, C-shaped aperture, optical tweezers, lab-on-a-chip, nanotechnology, C-shaped engraving, CSE, electron beam lithography, nanostructure