Central Michigan University 9 articles published in JoVE Biochemistry Bioluminescent Optogenetics 2.0: Harnessing Bioluminescence to Activate Photosensory Proteins In Vitro and In Vivo Emmanuel L. Crespo1,2, Andreas Bjorefeldt1, Mansi Prakash1, Ute Hochgeschwender1,2,3 1College of Medicine, Central Michigan University, 2Biochemistry, Cell and Molecular Biology Program, Central Michigan University, 3Neuroscience Program, Central Michigan University Bioluminescence-light emitted by a luciferase enzyme oxidizing a small-molecule substrate, a luciferin-can be harnessed to activate photosensory proteins, thereby adding another dimension to light stimulation and enabling the manipulation of a multitude of light-mediated functions in cells across temporal and spatial scales. Biology Quadruple-Checkerboard: A Modification of the Three-Dimensional Checkerboard for Studying Drug Combinations Christina Isber1, David L. Stockman2, Ziad Daoud2,3 1Faculty of Medicine and Medical Sciences, University of Balamand, 2Department of Clinical Microbiology, Michigan Health Clinics, 3College of Medicine, Central Michigan University This protocol describes how to study all possible combinations that can be obtained between four drugs in one single experiment. This method is based on the standard 96-well plate micro dilution assay and the calculation of fractional inhibitory concentrations (FICs) to evaluate the results. Neuroscience Labeling and Imaging of Amyloid Plaques in Brain Tissue Using the Natural Polyphenol Curcumin Panchanan Maiti1,2,3,4,5, Alexandra Plemmons1, Zackary Bowers2,3, Charles Weaver5, Gary Dunbar1,2,3,4 1 Curcumin is an ideal fluorophore for labeling and imaging of amyloid beta protein plaques in brain tissue due to its preferential binding to amyloid beta protein as well as its structural similarities with other traditional amyloid binding dyes. It can be used to label and image amyloid beta protein plaques more efficiently and inexpensively than traditional methods. Biology Preparation of Prokaryotic and Eukaryotic Organisms Using Chemical Drying for Morphological Analysis in Scanning Electron Microscopy (SEM) Madison A. Koon1, Khadijah Almohammed Ali1, Robert M. Speaker1, James P. McGrath1, Eric W. Linton1, Michelle L. Steinhilb1 1Department of Biology, Central Michigan University SEM analysis is an effective method to aid in species identification or phenotypic discrimination. This protocol describes the methods for examining specific morphological details of three representative types of organisms and would be broadly applicable to examining features of many organismal and tissue types. Genetics Reverse Transcription-Loop-mediated Isothermal Amplification (RT-LAMP) Assay for Zika Virus and Housekeeping Genes in Urine, Serum, and Mosquito Samples Sarah N. Bartolone1, Maya O. Tree2, Michael J. Conway2, Michael B. Chancellor1,3, Laura E. Lamb1,3 1Department of Urology, Beaumont Health System, 2Foundational Sciences, College of Medicine, Central Michigan University, 3Oakland University William Beaumont School of Medicine This protocol provides an efficient and low-cost method to detect Zika virus or control targets in human urine and serum samples or in mosquitoes by reverse transcription loop-mediated isothermal amplification (RT-LAMP). This method does not require RNA isolation and can be done within 30 min. Medicine Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling Michelle J. Veite-Schmahl1, Daniel P. Regan1, Adam C. Rivers1, Joseph F. Nowatzke1,2, Michael A. Kennedy1 1Department of Chemistry & Biochemistry, Miami University, 2College of Medicine, Central Michigan University This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling. Biochemistry Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues Lisa M. Meints1, Anne W. Poston1, Brent F. Piligian1, Claire D. Olson1, Katherine S. Badger2, Peter J. Woodruff2, Benjamin M. Swarts1 1Department of Chemistry and Biochemistry, Central Michigan University, 2Department of Chemistry, University of Southern Maine Trehalose analogues are emerging as important molecules for bio(techno)logical and biomedical applications. We describe an optimized protocol for enzymatically synthesizing and purifying trehalose analogues that is simple, efficient, fast, and environmentally friendly. Its application to the rapid production and administration of a probe for the detection of mycobacteria is demonstrated. Engineering Electrochemical Etching and Characterization of Sharp Field Emission Points for Electron Impact Ionization Tyler L. Van Well1, Matthew Redshaw1, Nadeesha D. Gamage1, R. M. Eranjan B. Kandegedara1 1Department of Physics, Central Michigan University A method for electrochemically etching field emission tips is presented. Etching parameters are characterized and the operation of the tips in field emission mode is investigated. Medicine Modeling and Simulations of Olfactory Drug Delivery with Passive and Active Controls of Nasally Inhaled Pharmaceutical Aerosols Xiuhua A. Si1, Jinxiang Xi2 1Department of Mechanical Engineering, California Baptist University, 2School of Engineering and Technology, Central Michigan University This manuscript reviews the modeling and simulations of different protocols to deliver medications to the olfactory region in image-based nasal airway models. Multiple software modules are used to develop the anatomically accurate nose model, generate computational mesh, simulate nasal airflows, and predict particle deposition at the olfactory region.