Loyola University Chicago View Institution's Website 10 articles published in JoVE Chemistry Characterizing Lewis Pairs Using Titration Coupled with In Situ Infrared Spectroscopy Carly S. Hanson1, James J. Devery1 1Department of Chemistry & Biochemistry, Loyola University Chicago Here, we present a method for the observation of solution interactions between Lewis acids and bases by employing in situ infrared spectroscopy as a detector for titration under synthetically relevant conditions. By examining solution interactions, this method represents a complement to X-ray crystallography, and provides an alternative to NMR spectroscopy. Developmental Biology Assay for Blood-brain Barrier Integrity in Drosophila melanogaster Matthew J. Davis1, Danielle Talbot1, Jennifer Jemc1 1Department of Biology, Loyola University Chicago Blood-brain barrier integrity is critical for nervous system function. In Drosophila melanogaster, the blood-brain barrier is formed by glial cells during late embryogenesis. This protocol describes methods to assay for blood-brain barrier formation and maintenance in D. melanogaster embryos and third instar larvae. Biology Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures David J. Rademacher1, Maleen Cabe2, Joanna C. Bakowska2 1Core Imaging Facility and Department of Microbiology and Immunology, Loyola University Chicago, 2Department of Molecular Pharmacology and Therapeutics, Loyola University Chicago We describe a comprehensive and practical protocol for fluorescence recovery after photobleaching experiments with live cells. Although the protocol was used to measure the mobility of yellow fluorescent protein-tagged p62 in aggresome-like induced structures, it can be applied to a variety of microscopy systems and fluorescent proteins. Neuroscience In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model Symantas Ragauskas1, Eva Kielczewski2, Joseph Vance2,3, Simon Kaja1,4, Giedrius Kalesnykas1 1Experimentica Ltd., 2Leica Microsystems, 3Spective LLC, 4Department of Ophthalmology, Stritch School of Medicine, Loyola University Chicago Here, we present the usefulness of longitudinal in vivo imaging in the follow-up of morphological changes of laser-induced choroidal neovascularization in mice. Medicine Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability Christina Wayman*1,2, Denise A. Duricki*1,2, Lisa A. Roy3, Barbara Haenzi1, Shi-Yen Tsai4, Gwendolyn Kartje4,5,6, John S. Beech7, Diana Cash2, Lawrence Moon1 1Department of Neuroimaging, James Black Centre, Institute of Psychiatry, King's College London, University of London, 3Institute of Neuroscience and Psychology, Wellcome Surgical Institute, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, 4Research Service, Edward Hines Jr. VA Hospital, 5Neurology Service, Edward Hines Jr. VA Hospital, 6Department of Molecular Pharmacology and Therapeutics, Neuroscience Research Institute, Loyola University Chicago, 7Department of Oncology, The Gray Institute for Radiation, Oncology and Biology, University of Oxford Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke. Biology Voltage and Calcium Dual Channel Optical Mapping of Cultured HL-1 Atrial Myocyte Monolayer Jiajie Yan*1, Justin K. Thomson*1, Weiwei Zhao1, Vladimir G. Fast2, Tong Ye3, Xun Ai1 1Department of Cell and Molecular Physiology, Loyola University Chicago, 2Department of Biomedical Engineering, University of Alabama at Birmingham, 3Department of Bioengineering, Clemson University This article describes the technique used to perform dual channel optical mapping in cultured HL-1 atrial cell monolayers. This unique protocol allows the simultaneous visualization of both calcium (Ca) and voltage (Vm) activity in the same area for the detailed detection and analysis of electrophysiological properties of culture monolayers. Biology A Sensitive and Specific Quantitation Method for Determination of Serum Cardiac Myosin Binding Protein-C by Electrochemiluminescence Immunoassay Diederik W.D. Kuster1, David Barefield1, Suresh Govindan1, Sakthivel Sadayappan1 1Department of Cell and Molecular Physiology, Loyola University Chicago Measuring biomarkers in complex biological samples is increasingly guiding clinical decision-making. We describe a highly sensitive method to simultaneously measure cardiac myosin binding protein-C, creatine kinase MB, and cardiac troponin I in serum samples from subjects with myocardial infarction and healthy control subjects. Biology Exploring Arterial Smooth Muscle Kv7 Potassium Channel Function using Patch Clamp Electrophysiology and Pressure Myography Lioubov I. Brueggemann1, Bharath K. Mani1, Jennifer Haick1, Kenneth L. Byron1 1Department of Molecular Pharmacology & Therapeutics, Loyola University Chicago Measurements of Kv7 (KCNQ) potassium channel activity in isolated arterial myocytes (using patch clamp electrophysiological techniques) in parallel with measurements of constrictor/dilator responses (using pressure myography) can reveal important information about the roles of Kv7 channels in vascular smooth muscle physiology and pharmacology. Biology A Semi-quantitative Approach to Assess Biofilm Formation Using Wrinkled Colony Development Valerie A. Ray1, Andrew R. Morris1, Karen L. Visick1 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. Neuroscience Determination of Mitochondrial Membrane Potential and Reactive Oxygen Species in Live Rat Cortical Neurons Dinesh C. Joshi1, Joanna C. Bakowska1 1Department of Molecular Pharmacology and Experimental Therapeutics, Loyola University Chicago We demonstrate application of the fluorescence indicator, TMRM, in cortical neurons to determine the relative changes in TMRM fluorescence intensity before and after application of a specific stimulus. We also show application of the fluorescence probe H2DCF-DA to assess the relative level of reactive oxygen species in cortical neurons.