University of Akron 9 articles published in JoVE Chemistry Depolymerizable Olefinic Polymers Based on Fused-Ring Cyclooctene Monomers Devavrat Sathe1, Junfeng Zhou1, Hanlin Chen1, Junpeng Wang1 1School of Polymer Science and Polymer Engineering, University of Akron Here, we describe protocols for the preparation of trans-cyclobutane fused cyclooctenes (tCBCO), their polymerization to prepare depolymerizable olefinic polymers, and the depolymerization of these polymers under mild conditions. Additionally, protocols for the preparation of depolymerizable networks and compression molding of rigid linear plastics based on this system are described. Biochemistry A Model Membrane Platform for Reconstituting Mitochondrial Membrane Dynamics Yifan Ge1, Sivakumar Boopathy1, Adam Smith2, Luke H. Chao1,3 1Department of Molecular Biology, Massachusetts General Hospital, 2Department of Chemistry, University of Akron, 3Department of Genetics, Harvard Medical School Mitochondrial fusion is an important homeostatic reaction underlying mitochondrial dynamics. Described here is an in vitro reconstitution system to study mitochondrial inner-membrane fusion that can resolve membrane tethering, docking, hemifusion, and pore opening. The versatility of this approach in exploring cell membrane systems is discussed. Biology A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT Janna M. Andronowski1, Reed A. Davis1, Caleb W. Holyoke2 1Department of Biology, The University of Akron, 2Department of Geosciences, The University of Akron We employed a geological (coring) sampling protocol to procure cortical bone specimens of uniform size for SRµCT experiments from the anterior aspect of human femora. This method is minimally destructive, efficient, results in cylindrical specimens that minimize imaging artifacts from irregular sample shapes and improves microarchitectural visualization and analysis. Bioengineering A Lab-On-A-Chip Platform for Stimulating Osteocyte Mechanotransduction and Analyzing Functional Outcomes of Bone Remodeling Sharon L. Truesdell1, Estee L. George1, Christopher C. Van Vranken1, Marnie M. Saunders1 1Department of Biomedical Engineering, The University of Akron Here, we present protocols for analyzing bone remodeling within a lab-on-a-chip platform. A 3D printed mechanical loading device can be paired with the platform to induce osteocyte mechanostransduction by deforming the cellular matrix. The platform can also be used to quantify bone remodeling functional outcomes from osteoclasts and osteoblasts (resorption/formation). Engineering Application of Design Aspects in Uniaxial Loading Machine Development Robert P. Thoerner1, Jonathan D. King1, Marnie M. Saunders1 1Department of Biomedical Engineering, University of Akron Here we present a protocol to develop a pure uniaxial loading machine. Critical design aspects are employed to ensure accurate and reproducible testing results. Neuroscience Where You Cut Matters: A Dissection and Analysis Guide for the Spatial Orientation of the Mouse Retina from Ocular Landmarks Katelyn B. Sondereker1, Maureen E. Stabio2, Jenna R. Jamil1, Matthew J. Tarchick1, Jordan M. Renna1 1Department of Biology, The University of Akron, 2Department of Cell and Developmental Biology, University of Colorado Denver This protocol provides a comprehensive dissection and analysis guide for the use of deep ocular landmarks, s-opsin immunohistochemistry, Retistruct, and custom code to accurately and reliably orient the isolated mouse retina in anatomical space. Bioengineering Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing Stephanie Lemmo Ham1, Ehsan Atefi1, Darcy Fyffe1, Hossein Tavana1 1Department of Biomedical Engineering, The University of Akron A protocol for robotic printing of cancer cell spheroids in a high throughput 96-well plate format using an aqueous two-phase system is presented. Bioengineering Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration Aleesha M. McCormick1, Natalie A. Jarmusik1, Elizabeth J. Endrizzi1, Nic D. Leipzig1 1Department of Chemical and Biomolecular Engineering, University of Akron Developing biotinylatable fusion proteins has many potential applications in various fields of research. Recombinant protein engineering is a straight forward procedure that is cost-effective, providing high yields of custom-designed proteins. Bioengineering Characteristics of Precipitation-formed Polyethylene Glycol Microgels Are Controlled by Molecular Weight of Reactants Susan Thompson1, Jessica Stukel1, Abrar AlNiemi1,2, Rebecca Kuntz Willits1 1Department of Biomedical Engineering, The University of Akron, 2 This work describes the formation of poly(ethylene glycol) (PEG) microgels via a photopolymerized precipitation reaction. Increasing the PEG molecular weight increased microgel diameter and swelling ratio. Simple adaptations to the PEG microgel precipitation reaction are explored for future applications of microgels as drug delivery vehicles and tissue engineering scaffolds.