1Focal Area Infection Biology, Biozentrum of the University of Basel, 2Proteomics Core Facility, Biozentrum of the University of Basel
The ubiquitous second messenger c-di-GMP controls growth and behavior of many bacteria. We have developed a novel Capture Compound Mass Spectrometry based technology to biochemically identify and characterize c-di-GMP binding proteins in virtually any bacterial species.
Published March 29, 2015. Keywords: Chemistry, Capture compound, photoactivable crosslinker, mass spectrometry, c-di-GMP effector, EAL, GGDEF, PilZ, Pseudomonas aeruginosa
1National Exposure Research Laboratory, U.S Environmental Protection Agency, 2Technical Support Center, Office of Ground Water and Drinking Water, U.S Environmental Protection Agency
EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.
Published March 28, 2015. Keywords: Environmental Sciences, enteric virus, environmental microbiology, water, virus occurrence, electropositive cartridge filters, sample collection
1Neural Engineering Center, Department of Biomedical Engineering, Case Western Reserve University
We have developed an in vitro unfolded hippocampus which preserves CA1-CA3 array of neurons. Combined with the penetrating micro-electrode array, neural activity can be monitored in both the longitudinal and transverse orientations. This method provides advantages over hippocampal slice preparations as the propagation in the entire hippocampus can be recorded simultaneously.
Published March 27, 2015. Keywords: Neuroscience, Penetrating micro-electrode array (PMEA), unfolded intact hippocampus, neural activity propagation, neural signal mapping, flat pyramidal cell sheet, unfolded hippocampus placement
1Department of Orthopaedic Surgery, University of California Los Angeles, 2Department of Bioengineering, University of California Los Angeles
Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.
Published March 26, 2015. Keywords: Bioengineering, Anterior cruciate ligament, tissue engineering, animal model, biodegradable scaffold, rat, knee
1Department of Clinical Neurosciences, Hotchkiss Brain Institute at University of Calgary, 2Department of Oncology, Hotchkiss Brain Institute at University of Calgary
Demyelinating diseases can be modeled in animals by focal application of lysolecithin into the CNS. A single injection of lysolecithin into mouse spinal cord produces a lesion that spontaneously repairs over time. The goal is to study factors involved in de- and remyelination, and to test agents for enhancing repair.
Published March 26, 2015. Keywords: Neuroscience, demyelination, remyelination, lysolecithin, spinal cord, oligodendrocyte, myelin, multiple sclerosis
JoVE Developmental Biology
1Feinberg Cardiovascular Research Institute, Northwestern University, 2Cardiovascular Research Institute, University of California, San Francisco
Mutations that lead to congenital heart defects benefit from in vivo investigation of cardiac structure during development, but high-resolution structural studies in the mouse embryonic heart are technically challenging. Here we present a robust immunofluorescence and image analysis method to assess cardiomyocyte-specific structures in the developing mouse heart.
Published March 26, 2015. Keywords: Developmental Biology, Immunofluorescence, mouse, embryonic heart, cardiomyocyte, development, sarcomere, intercalated disc, costamere, s-α-actinin, cryosection
1Departments of Chemistry and Cell and Molecular Biology, The Scripps Research Institute, 2Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University
Zinc-finger domains are intrinsically cell-permeable and capable of mediating protein delivery into a broad range of mammalian cell types. Here, a detailed step-by-step protocol for implementing zinc-finger technology for intracellular protein delivery is presented.
Published March 25, 2015. Keywords: Molecular Biology, protein delivery, cell-penetrating peptide, zinc-finger, protein transduction domain, chemical biology, molecular biology
1Biomedical Engineering Department, Saint Louis University
Here, a method that enables quick, efficient, and inexpensive preparation of polyacrylamide gels in a multiwell plate format is described. The method does not require any specialized equipment and could be easily adopted by any research laboratory. It would be particularly useful in research focused on understanding stiffness-dependent cell responses.
Published March 25, 2015. Keywords: Bioengineering, Multiwell, substrate stiffness, drug screening, polyacrylamide, Young’s modulus, high-throughput
JoVE Immunology and Infection
1Center for Biologics Evaluation and Research, US Food and Drug Administration, 2Center for Drug Evaluation and Research, US Food and Drug Administration
This protocol describes a high-throughput qRT-PCR assay for the analysis of type I and III IFN expression signatures. The assay discriminates single base pair differences between the highly similar transcripts of these genes. Through batch assembly and robotic pipetting, the assays are consistent and reproducible.
Published March 24, 2015. Keywords: Immunology, Interferon, Innate Immunity, qRT-PCR Assay, Probes, Primers, Automated Pipetting
JoVE Immunology and Infection
1Department of Medicine III, RWTH University-Hospital Aachen, 2IZKF Aachen Core Facility "Two-Photon Imaging", RWTH University-Hospital Aachen, 3Institute for Laboratory Animal Science & Experimental Surgery, RWTH Aachen University, 4Institute for Pharmacology, RWTH University-Hospital Aachen
Stable intravital high-resolution imaging of immune cells in the liver is challenging. Here we provide a highly sensitive and reliable method to study migration and cell-cell-interactions of immune cells in mouse liver over long periods (about 6 hours) by intravital multiphoton laser scanning microscopy in combination with intensive care monitoring.
Published March 24, 2015. Keywords: Immunology, intravital imaging, TPLSM, two-photon microscopy, liver, migration, microscopy, leukocyte traffic, inflammation