1Department of Ophthalmology, University of Basel, 2Department of Biomedicine, University of Basel
Comet assay measures DNA breaks, induced by different factors. If all factors (except oxidative stress) causing DNA damage are kept constant, the amount of DNA damage is a good indirect parameter of oxidative stress. The goal of this protocol is to use comet assay for indirect measurement of oxidative stress.
Published May 22, 2015. Keywords: Molecular Biology, Comet assay analysis, Single-cell gel electrophoresis, DNA breaks, Oxidative stress
1Department of Mechanical Engineering, Massachusetts Institute of Technology, 2Laboratory for Manufacturing and Productivity, Massachusetts Institute of Technology, 3School of Engineering and Applied Sciences, Harvard University, 4Department of Materials Science and Engineering, Massachusetts Institute of Technology, 5Department of Chemistry & Chemical Biology, Harvard University
Tin sulfide (SnS) is a candidate material for Earth-abundant, non-toxic solar cells. Here, we demonstrate the fabrication procedure of the SnS solar cells employing atomic layer deposition, which yields 4.36% certified power conversion efficiency, and thermal evaporation which yields 3.88%.
Published May 22, 2015. Keywords: Engineering, Solar cells, thin films, thermal evaporation, atomic layer deposition, annealing, tin sulfide
1Institut de Physique du Globe de Paris, Institut Universitaire de France, Université Paris Diderot, 2INSERM (Institut National de la Santé et de la Recherche Médicale), U1148: Laboratory for Vascular Translational Science, Université Paris Diderot
We present the technique to measure with high precision zinc isotope ratios in mouse organs.
Published May 22, 2015. Keywords: Chemistry, Zinc, isotopes, MC-ICP-MS, chromatography, mice, purification
JoVE Immunology and Infection
1Department of Obstetrics & Gynecology, Wayne State University School of Medicine, 2School of Paediatrics and Reproductive Health, Research Centre for Reproductive Health, the Robinson Research Institute, The University of Adelaide, 3Department of Immunology & Microbiology, Wayne State University School of Medicine, 4Perinatology Research Branch, NICHD/NIH/DHHS
Described herein is a protocol to isolate and analyze the infiltrating leukocytes of tissues at the maternal-fetal interface (uterus, decidua, and placenta) of mice. This protocol maintains the integrity of most cell surface markers and yields enough viable cells for downstream applications including flow cytometry analysis.
Published May 21, 2015. Keywords: Immunology, Decidua, Dissociation, Isolation, Leukocytes, Myometrium, Placenta, Pregnancy, Uterus
JoVE Immunology and Infection
1Perinatology Research Branch, NICHD/NIH/DHHS, 2Department of Obstetrics and Gynecology, University of Michigan, 3Department of Epidemiology and Biostatistics, Michigan State University, 4Department of Molecular Obstetrics and Genetics, Wayne State University, 5Department of Obstetrics and Gynecology, Wayne State University School of Medicine, 6Department of Immunology and Microbiology, Wayne State University School of Medicine
Described herein is a protocol to isolate and further study the infiltrating leukocytes of the decidua basalis and decidua parietalis - the human maternal-fetal interface. This protocol maintains the integrity of cell surface markers and yields enough viable cells for downstream applications as proven by flow cytometry analysis.
Published May 21, 2015. Keywords: Immunology, Accutase, Decidua Basalis, Decidua Parietalis, Flow Cytometry, Immunophenotyping, Pregnancy
JoVE Immunology and Infection
1Department of Oral Microbiology and Immunology, School of Dentistry and Dental Research Institute, Seoul National University
Here a method to localize bacteria within paraffin-embedded tissues using DIG-labeled 16S rRNA-targeting DNA probes has been described. This protocol can be applied to study the role of bacteria in various diseases such as periodontitis, cancers, and inflammatory immune diseases.
Published May 21, 2015. Keywords: Immunology, periodontology, oral microbiology, in situ hybridization, 16S rRNA, bacteria, paraffin-embedded tissue, species-specific, universal
JoVE Developmental Biology
1Department of Developmental Biology, McGowan Institute for Regenerative Medicine, University of Pittsburgh
To help assess the molecular mechanisms underlying zebrafish biliary-driven liver regeneration, we established a liver injury model in which the nitroreductase-expressing hepatocytes are genetically ablated upon metronidazole treatment. In this protocol, we describe how to adeptly manipulate, monitor and analyze hepatocyte ablation and biliary-driven liver regeneration.
Published May 20, 2015. Keywords: Developmental Biology, nitroreductase, metronidazole, biliary epithelial cells, hepatocytes, zebrafish, liver regeneration, cell ablation, transgenic
1Department of Baker Institute for Animal Health, Cornell University, 2Department of Molecular Biology and Genetics, Cornell University, 3Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School
Here, we document the use of the soft agar colony formation assay to test the effects of a peptidylarginine deiminase (PADI) enzyme inhibitor, BB-Cl-amidine, on breast cancer tumorigenicity in vitro.
Published May 20, 2015. Keywords: Medicine, Peptidylarginine deiminase enzymes, breast cancer, soft agar assay, cellular transformation, cancer therapies
1Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center
The goal of this protocol is to study breast tumorigenesis. With this technique, mouse mammary tumors are removed and primary cells are prepared from tumors. A lung extraction protocol is included for studying lung metastasis. Furthermore, another protocol for analyzing mouse embryonic fibroblasts from the mouse embryo is included.
Published May 20, 2015. Keywords: Medicine, Tumor, breast, lung, primary, MEF, embryo, fibroblasts, cancer, cell, mouse
1Department of Physiology and Neurobiology, University of Connecticut, 2Department of Anatomic Pathology and Laboratory Medicine, University of Connecticut Health Center
Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain’s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.
Published May 19, 2015. Keywords: Neuroscience, Aging, ventriculomegaly, lateral ventricles, MRI, ependymal cells, glial scarring