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24 ARTICLES PUBLISHED IN JoVE

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Biology

AC Electrokinetic Phenomena Generated by Microelectrode Structures
Robert Hart 1, Jonghyun Oh 2, Jorge Capurro 2, Hongseok (Moses) Noh 2
1Biomedical Engineering, Science and Health Systems, Drexel University, 2Mechanical Engineering and Mechanics, Drexel University

Manipulating fluids and suspended particles in the micro- and nano-scale is becoming more of a reality as enabling technologies, like AC electrokinetics, continue to develop. Here, we discuss the physics behind AC electrokinetics, how to fabricate these devices and how to interpret the experimental observations.

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Biology

Optimization of the Ugi Reaction Using Parallel Synthesis and Automated Liquid Handling
Jean-Claude Bradley 1, Khalid Baig Mirza 1, Tom Osborne 2, Antony Wiliams 3, Kevin Owens 1
1Department of Chemistry, Drexel University, 2Mettler-Toledo, 3Chemspider

The Ugi reaction has proved to be a convenient way to quickly create diverse libraries of compounds. It involves the reaction of an amine, an aldehyde, a carboxylic acid and an isonitrile typically in methanol at room temperature. In this video, we utilize a 48-slot Mettler-Toledo MiniBlock equipped with filtration tubes and a Mettler-Toledo MiniMapper automated liquid handler was used to deliver the reagents and solvent. The parameters of interest were the concentration, the solvent composition and the excess of some of the reagents.

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Neuroscience

Using MazeSuite and Functional Near Infrared Spectroscopy to Study Learning in Spatial Navigation
Hasan Ayaz 1, Patricia A. Shewokis 1,2, Adrian Curtin 1, Meltem Izzetoglu 1, Kurtulus Izzetoglu 1, Banu Onaral 1
1School of Biomedical Engineering, Science and Health Systems, Drexel University, 2College of Nursing and Health Professions, Drexel University

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments. Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables noninvasive and portable monitoring of cerebral blood oxygenation changes. This paper summarizes collective use of MazeSuite and fNIR within a cognitive processing learning paradigm.

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Chemistry

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
Nathaniel W. Snyder 1, Maya Khezam 1, Clementina A. Mesaros 1, Andrew Worth 1, Ian A. Blair 1
1Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania

Untargeted metabolomics provides a hypothesis generating snapshot of a metabolic profile. This protocol will demonstrate the extraction and analysis of metabolites from cells, serum, or tissue. A range of metabolites are surveyed using liquid-liquid phase extraction, microflow ultraperformance liquid chromatography/high-resolution mass spectrometry (UPLC-HRMS) coupled to differential analysis software.

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Medicine

Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury
R. Vivian Allahyari 1, A. Denise R. Garcia 1
1Department of Biology, Drexel University

This article describes a detailed protocol to produce a forebrain stab injury in adult mice. The stab injury induces severe reactive gliosis and glial scar formation which can be subsequently examined by standard immunohistochemistry methods.

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Bioengineering

A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities
Jacob R. Price 1, Wen K. Shieh 2, Christopher M. Sales 1
1Civil, Architectural, and Environmental Engineering, Drexel University, 2Chemical and Biomolecular Engineering, University of Pennsylvania

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Here, the HDBR is successfully applied in a photobioreactor (PBR) configuration for the study of nitrogen metabolism by a mixed high density algal community.

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Bioengineering

Synthesis of Thermogelling Poly(N-isopropylacrylamide)-graft-chondroitin Sulfate Composites with Alginate Microparticles for Tissue Engineering
Thomas R. Christiani 1, Katelynn Toomer 2, Joseph Sheehan 2, Angelika Nitzl 2, Amanda Branda 2, Elizabeth England 2, Pamela Graney 3, Cristina Iftode 2, Andrea J. Vernengo 1
1Department of Chemical Engineering, Rowan University, 2Department of Biological Sciences, Rowan University, 3Department of Biomedical Engineering, Drexel University

An injectable tissue engineering scaffold composed of poly(N-isopropylacrylamide)-graft-chondroitin sulfate (PNIPAAm-g-CS)-containing alginate microparticles was prepared. The adhesive strength, swelling properties and in vitro biocompatibility are analyzed in this study. The characterization techniques developed here may be applicable to other thermogelling systems.

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Education

LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism
Andrew J. Worth *1,2, Dylan M. Marchione *2,3, Robert C. Parry 1,2, Qingqing Wang 2,3, Kevin P. Gillespie 2,3, Noelle N. Saillant 4, Carrie Sims 4, Clementina Mesaros 1,2, Nathaniel W. Snyder 5, Ian A. Blair 1,2
1Center for Cancer Pharmacology, University of Pennsylvania, 2Center for Excellence in Environmental Toxicology, University of Pennsylvania, 3Penn SRP and Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania, 4Division of Traumatology, Department of Surgery, Critical Care and Acute Care Surgery, University of Pennsylvania, 5A.J. Drexel Autism Institute, Drexel University

Here we show isolated human platelets can be used as an accessible ex vivo model to study metabolic adaptations in response to the complex I inhibitor rotenone. This approach employs isotopic tracing and relative quantification by liquid chromatography-mass spectrometry and can be applied to a variety of study designs.

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Environment

Comparison of Scale in a Photosynthetic Reactor System for Algal Remediation of Wastewater
Kaitlyn D. Sniffen 1, Christopher M. Sales 1, Mira S. Olson 1
1Civil, Architectural, and Environmental Engineering, Drexel University

An experimental methodology is presented to compare the performance of small (100 L) and large (1,000 L) scale reactors designed for algae remediation of landfill wastewater. System characteristics, including surface area to volume ratio, retention time, biomass density, and wastewater feed concentrations, can be adjusted based on application.

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Neuroscience

Anatomically Inspired Three-dimensional Micro-tissue Engineered Neural Networks for Nervous System Reconstruction, Modulation, and Modeling
Laura A. Struzyna *1,2,3, Dayo O. Adewole *1,2,3, Wisberty J. Gordián-Vélez 1,2,3, Michael R. Grovola 2,3, Justin C. Burrell 2,3, Kritika S. Katiyar 2,3,4, Dmitriy Petrov 2,3, James P. Harris 2,3, D. Kacy Cullen 2,3
1Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, 2Center for Brain Injury & Repair, Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, 3Center for Neurotrauma, Neurodegeneration & Restoration, Michael J. Crescenz Veterans Affairs Medical Center, 4School of Biomedical Engineering, Drexel University

This manuscript details the fabrication of micro-tissue engineered neural networks: three-dimensional micron-sized constructs comprised of long aligned axonal tracts spanning aggregated neuronal population(s) encased in a tubular hydrogel. These living scaffolds can serve as functional relays to reconstruct or modulate neural circuitry or as biofidelic test-beds mimicking gray-white matter neuroanatomy.

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JoVE Journal

Three-dimensional Tissue Engineered Aligned Astrocyte Networks to Recapitulate Developmental Mechanisms and Facilitate Nervous System Regeneration
Kritika S. Katiyar *1,2,3, Carla C. Winter *1,2,4, Wisberty J. Gordián-Vélez 1,2,4, John C. O'Donnell 1,2, Yeri J. Song 1,5, Nicole S. Hernandez 1,5, Laura A. Struzyna 1,2,4, D. Kacy Cullen 1,2,5
1Center for Brain Injury & Repair, Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, 2Center for Neurotrauma, Neurodegeneration & Restoration, Michael J. Crescenz Veterans Affairs Medical Center, 3School of Biomedical Engineering, Drexel University, 4Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, 5Neuroscience Graduate Group, Perelman School of Medicine, University of Pennsylvania

We showcase the development of self-assembled, three-dimensional bundles of longitudinally aligned astrocytic somata and processes within a novel biomaterial encasement. These engineered "living scaffolds", exhibiting micron-scale diameter yet extending centimeters in length, may serve as test-beds to study neurodevelopmental mechanisms or facilitate neuroregeneration by directing neuronal migration and/or axonal pathfinding.

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Behavior

Calculating Heart Rate Variability from ECG Data from Youth with Cerebral Palsy During Active Video Game Sessions
Corey Landis 1, Margaret E. O'Neil 2,3, Andrew Finnegan 2,3, Patricia A. Shewokis 1,3,4
1School of Biomedical Engineering, Science and Health Systems, Drexel University, 2Department of Physical Therapy and Rehabilitation Sciences, Drexel University, 3College of Nursing and Health Professions, Drexel University, 4Department of Nutrition Sciences, Drexel University

This protocol describes a method for calculating Heart Rate Variability (HRV) from electrocardiogram (ECG) waveforms. Waveforms from continuous heart rate (HR) recordings during active video game (AVG) sessions were used to measure the aerobic performance of youth with cerebral palsy (CP).

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Bioengineering

Methods for In Vivo Biomechanical Testing on Brachial Plexus in Neonatal Piglets
Anita Singh 1, Rachel Magee 1, Sriram Balasubramanian 2
1Department of Biomedical Engineering, Widener University, 2School of Biomedical Engineering, Science and Health Systems, Drexel University

Presented here are methods to perform in vivo biomechanical testing on brachial plexus in a neonatal piglet model.

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Bioengineering

Fabrication of Ti3C2 MXene Microelectrode Arrays for In Vivo Neural Recording
Nicolette Driscoll 1,2,3, Kathleen Maleski 4,5, Andrew G. Richardson 2,6, Brendan Murphy 1,2,3, Babak Anasori 4,5, Timothy H. Lucas 2,6, Yury Gogotsi 4,5, Flavia Vitale 1,2,3,7,8
1Department of Bioengineering, University of Pennsylvania, 2Center for Neuroengineering and Therapeutics, University of Pennsylvania, 3Corporal Michael J. Crescenz Veterans Affairs Medical Center, 4Department of Materials Science and Engineering, Drexel University, 5A.J. Drexel Nanomaterials Institute, Drexel University, 6Department of Neurosurgery, University of Pennsylvania, 7Department of Neurology, University of Pennsylvania, 8Department of Physical Medicine and Rehabilitation

We describe here a method for fabricating Ti3C2 MXene microelectrode arrays and utilizing them for in vivo neural recording.

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Genetics

Inducing Hairy Roots by Agrobacterium rhizogenes-Mediated Transformation in Tartary Buckwheat (Fagopyrum tataricum)
Yaolei Mi 1,2, Zhihui Zhu 3, Guangtao Qian 3, Yu Li 4, Xiangxiao Meng 1, Jianping Xue 3, Qingfu Chen 5, Wei Sun 1, Yuhua Shi 1,2
1Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, 2Artemisinin Research Center, China Academy of Chinese Medical Sciences, 3College of Life Science, Huaibei Normal University, 4Economic Crop Research Institute Sichuan Academy of Agriculture Sciences, 5Research Center of Buckwheat Industry Technology, Guizhou Normal University

We describe a method of inducing hairy roots by Agrobacterium rhizogenes-mediated transformation in Tartary buckwheat (Fagopyrum tataricum). This can be used to investigate gene functions and production of secondary metabolites in Tartary buckwheat, be adopted for any genetic transformation, or used for other medicinal plants after improvement.

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Environment

Developing a Virtual Reality Video Game to Simulate Rip Currents
Jase Bernhardt 1, Gregory Dusek 2, Arielle Hesse 3, William Santos 4, Thomas Jennings 4, Alexander Smiros 4, Antoinette Montes 1
1Department of Geology, Environment and Sustainability, Hofstra University, 2National Ocean Service, National Oceanographic and Atmospheric Administration, 3Department of Sociology, Drexel University, 4EdTech, Hofstra University

Rip currents are among the deadliest meteorological hazards in the United States. In order to demonstrate the proper actions to take when caught in a rip current in a memorable and engaging way, a virtual reality video game is developed.

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Bioengineering

Direct Bioprinting of 3D Multicellular Breast Spheroids onto Endothelial Networks
Swathi Swaminathan 1, Alisa Morss Clyne 2
1Department of Biology, Drexel University, 2Fischell Department of Bioengineering, University of Maryland

The goal of this protocol is to directly bioprint breast epithelial cells as multicellular spheroids onto pre-formed endothelial networks to rapidly create 3D breast-endothelial co-culture models which can be used for drug screening studies.

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Medicine

Virtual Reality Tools for Assessing Unilateral Spatial Neglect: A Novel Opportunity for Data Collection
Peter J. Schwab 1,2, Alex Miller 2, Ann-Marie Raphail 3, Ari Levine 2, Christopher Haslam 2, H. Branch Coslett 1,2, Roy H. Hamilton 1,2,4
1Department of Neurology, University of Pennsylvania, 2Laboratory for Cognition and Neural Stimulation, University of Pennsylvania, 3Department of Psychology, Drexel University, 4Department of Physical Medicine and Rehabilitation, University of Pennsylvania

The goal was to design, build, and pilot a novel virtual reality task to detect and characterize unilateral spatial neglect, a syndrome affecting 23-46% of acute stroke survivors, expanding the role of virtual reality in the study and management of neurologic disease.

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Bioengineering

Contrast-Enhanced Subharmonic Aided Pressure Estimation (SHAPE) Using Ultrasound Imaging with a Focus on Identifying Portal Hypertension
Flemming Forsberg 1, Ipshita Gupta 1,2, Priscilla Machado 1, Colette M. Shaw 1, Jonathan M. Fenkel 3, Kirk Wallace 4, John R. Eisenbrey 1
1Department of Radiology, Thomas Jefferson University, 2School of Biomedical Engineering, Sciences and Health Systems, Drexel University, 3Department of Medicine, Division of Gastroenterology and Hepatology, Thomas Jefferson University, 4GE Global Research

A protocol for noninvasively estimating ambient pressures utilizing subharmonic ultrasound imaging of infused contrast microbubbles (following appropriate calibration) is described with examples from human patients with chronic liver disease.

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Bioengineering

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting
Tiankun Liu *1,2, Yongchun Shao *1,2, Zitong Wang 1,2, Yuqiu Chen 1,2, Yuan Pang 1,2, Ding Weng 1,2, Wei Sun 1,2,3
1Biomanufacturing Center, Dept. of Mechanical Engineering, Tsinghua University, 2Biomanufacturing and Rapid Forming Technology Key Laboratory of Beijing, 3Department of Mechanical Engineering, Drexel University

Presented here is a mild 3D printing technique driven by alternating viscous-inertial forces to enable the construction of hydrogel microcarriers. Homemade nozzles offer flexibility, allowing easy replacement for different materials and diameters. Cell binding microcarriers with a diameter of 50-500 µm can be obtained and collected for further culturing.

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Neuroscience

Measurement of Oxygen Consumption Rate in Acute Striatal Slices from Adult Mice
Lianteng Zhi 1, Jingyu Zhao 1, David Jaffe 1, Yuanxin Chen 1, Ninghan Wang 1,2, Qi Qin 1, Erin L. Seifert 3, Chenjian Li 4, Hui Zhang 1
1Department of Neuroscience, Thomas Jefferson University, 2School of Biomedical Engineering, Drexel University, 3Department of Pathology, MitoCare Center, Thomas Jefferson University, 4School of Life Sciences, Peking University

Oxygen consumption rate (OCR) is a common proxy for mitochondrial function and can be used to study different disease models. We developed a new method using a Seahorse XF analyzer to directly measure the OCR in acute striatal slices from adult mice that is more physiologically relevant than other methods.

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Bioengineering

Investigating Stress-relaxation and Failure Responses in the Trachea
Anita Singh 1, Tanmay Majmudar 2,3, Adi Iyer 4, Diya Iyer 4, Sriram Balasubramanian 3
1Department of Biomedical Engineering, Widener University, 2Drexel University College of Medicine, 3School of Biomedical Engineering, Science and Health Systems, Drexel University, 4Rosetree Media School District

The present protocol determines the tensile stress-relaxation and failure properties of porcine tracheae. Results from such methods can help improve the understanding of the viscoelastic and failure thresholds of the trachea and help advance the capabilities of computational models of the pulmonary system.

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Bioengineering

Direct Linear Transformation for the Measurement of In-Situ Peripheral Nerve Strain During Stretching
Virginia Orozco 1, Sriram Balasubramanian 1, Anita Singh 2
1School of Biomedical Engineering, Science and Health Systems, Drexel University, 2Bioengineering Department, Temple University

This protocol implements a stereo-imaging camera system calibrated using direct linear transformation to capture three-dimensional in-situ displacements of stretched peripheral nerves. By capturing these displacements, strain induced at varying degrees of stretch can be determined informing the stretch injury thresholds that can advance the science of stretch-dependent nerve repair.

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Three-Dimensional Mapping of the Rotation of Interactive Virtual Objects with Eye-Tracking Data

Three-Dimensional Mapping of the Rotation of Interactive Virtual Objects with Eye-Tracking Data
Rodrigo A. Consoli 1, Eduardo Colli 2, Raimundo da Silva Soares Jr. 3,4, João R. Sato 3, Guilherme A. Marson 1
1Graduate Program in Chemistry, University of São Paulo, Institute of Chemistry, 2Graduate Program in Applied Mathematics, University of São Paulo, Institute of Mathematics and Statistics, 3Graduate Program in Neuroscience and Cognition, Federal University of ABC, Center of Mathematics, Computing and Cognition, 4School of Biomedical Engineering, Science and Health Systems, Drexel University

We have developed a simple, customizable, and efficient method for recording quantitative processual data from interactive spatial tasks and mapping these rotation data with eye-tracking data.

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