1Discipline of Biomedical Science, School of Medical Sciences, Sydney Medical School, University of Sydney, 2School of Biomedical Sciences and Pharmacy, University of Newcastle
Analysis of vestibular hair cell function is complicated by their location deep within the hardest part of the skull, the petrous temporal bone. Most functional hair cell studies have used acutely isolated hair cells. Here we describe a semi-intact preparation of mouse vestibular epithelium for electrophysiological and two-photon microscopy studies.
Published June 13, 2013. Keywords: Neurobiology, Neuroscience, Cellular Biology, Molecular Biology, Biomedical Engineering, Anatomy, Physiology, Surgery, Vestibular, Hair cells, Epithelium, two-photon microscopy, isolated, semi-intact, electrophysiology, electroporation, microscopy, tissue, isolation, animal model
1Department of Microbiology and Immunology, Loyola University Medical Center
We provide a simple, semi-quantitative method to investigate biofilm formation in vitro. This method takes advantage of the Zeiss stemi 2000-C Dissecting Microscope (with camera attachment) to monitor both the timing and pattern of biofilm formation, as assessed by the development of wrinkled colonies.
Published June 7, 2012. Keywords: Microbiology, Immunology, Biofilm, wrinkled colony, rugose, Vibrio fischeri, Zeiss stemi, dissecting microscope, marine biology
1Neurosurgery, Baylor College of Medicine, 2Michael E. DeBakey Veterans Affairs Medical Center, 3Molecular & Cellular Biology, Baylor College of Medicine
We demonstrate the assembly and application of a molecular-scale device powered by a topoisomerase protein. The construct is a bio-molecular sensor which labels two major types of DNA breaks in tissue sections by attaching two different fluorophores to their ends.
Published January 11, 2012. Keywords: Bioengineering, molecular machine, bio-nanotechnology, 5'OH DNA breaks, 5'PO4 DNA breaks, apoptosis labeling, in situ detection, vaccinia topoisomerase I, DNA breaks, green nanotechnology
1Department of Molecular & Human Genetics, Baylor College of Medicine (BCM), 2Precisionary Instruments Inc., 3Departments of Molecular & Human Genetics and Neuroscience, Baylor College of Medicine (BCM), 4Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital
Here we describe a rapid and simple method to image fluorescently labeled cells in semi-thick brain slices. By fixing, slicing, and optically clearing brain tissue we describe how standard epifluorescent or confocal imaging can be used to visualize individual cells and neuronal networks within intact nervous tissue.
Published July 26, 2011. Keywords: Neuroscience, nervous tissue, neurons, confocal, epifluorescent, imaging, clearing, fluorescent proteins
1Development and Aging Program, The Sanford Burnham Institute for Medical Research, 2Cardiac Electrophysiology Group, Dept. of Physiology, Anatomy and Genetics, The Sanford Burnham Institute for Medical Research, 3Biology Department and Heart Institute, San Diego State University
We have developed a Semi-automated Optical Heartbeat Analysis method (SOHA) for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts. We demonstrate the application of our methodology to the analysis of heart function in fruit fly and embryonic mouse hearts.
Published September 16, 2009. Keywords: Physiology, Drosophila, zebrafish, mouse, heart, myosin, dilated, restricted, cardiomyopathy, KCNQ, movement detection
1Department of Pharmacology and Toxicology, University of Alabama at Birmingham
Described here is a protocol for semi-automated imaging of tissue-specific fluorescence in zebrafish embryos.
Published May 17, 2014. Keywords: Developmental Biology, zebrafish, Imaging, fluorescence microscopy, estrogen, developmental biology, endocrine disrupting compounds
1Department of Plant Biology, University of California Davis
Demonstration of key methods for high throughput leaf measurements. These methods can be used to accelerate leaf phenotyping when studying many plant mutants or otherwise screening plants by leaf phenotype.
Published January 21, 2013. Keywords: Plant Biology, Cellular Biology, Molecular Biology, Physiology, Computer Science, Arabidopsis, Arabidopsis thaliana, leaf shape, shade avoidance, ImageJ, LeafJ, petiole, touch-screen tablet, phenotyping, phenomics
1Department of Complexity Science and Engineering, Graduate School of Frontier Sciences, The University of Tokyo, 2Department of Physics, Graduate School of Science, The University of Tokyo
Here we describe a protocol for optogenetic manipulation of motoneuronal activity while monitoring changes in motor output (muscle contraction) in semi-intact Drosophila larvae using lasers within a conventional confocal microscope. This technique enables researchers to achieve local perturbation of neural activity in a few neuromeres to elucidate the dynamics of motor circuits.
Published July 4, 2013. Keywords: Neuroscience, Molecular Biology, Neurobiology, Developmental Biology, Bioengineering, Cellular Biology, Motor Neurons, Neurosciences, Drosophila, Optogenetics, Channelrhodopsin-2, Halorhodopsin, laser, confocal microscopy, animal model
1Whitehead Institute for Biomedical Research, 2Department of Biology, MIT - Massachusetts Institute of Technology, 3Howard Hughes Medical Institute
SDD-AGE is a useful technique for the detection and characterization of amyloid-like polymers in cells. Here we demonstrate an adaptation that makes this technique amenable to large-scale applications.
Published July 16, 2008. Keywords: Basic Protocols, biochemistry, SDD-AGE, amyloid, prion, aggregate
JoVE Application Notes
A demonstration of the advantages in speed, ease of use, and transfer efficiency of the Trans-Blot Turbo transfer system compared to conventional wet tank and semi-dry transfer methods.
Published July 15, 2011. Keywords: Protein transfer, turbo, rapid transfer, blotting, western blot, antigen, antibody, tank transfer, semi-dry, chemiluminescent, membrane