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Saint Louis University

14 ARTICLES PUBLISHED IN JoVE

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Biology

Determination of DNA Methylation of Imprinted Genes in Arabidopsis Endosperm
Matthew Rea 1, Ming Chen 1, Shan Luan 1, Drutdaman Bhangu 1, Max Braud 1, Wenyan Xiao 1
1Department of Biology, Saint Louis University

Imprinting is a phenomenon in plant and mammal reproduction. DNA methylation plays an important role in mechanisms of imprinting. Isolating endosperm and determining methylation status of imprinted genes in Arabidopsis can be difficult. In this protocol, we describe how to isolate endosperm and determine methylation by bisulfite sequencing.

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Bioengineering

Mechanical Testing of Mouse Carotid Arteries: from Newborn to Adult
Mazyar Amin 1, Victoria P. Le 1, Jessica E. Wagenseil 1
1Department of Biomedical Engineering, Saint Louis University

Passive mechanical testing of mouse carotid arteries is described, with special consideration for adapting to different specimen ages. The procedures include determining the in vivo length of the artery, mounting it in a pressure myograph, recording data, measuring the unloaded dimensions and analyzing the resulting data.

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Bioengineering

Measuring Left Ventricular Pressure in Late Embryonic and Neonatal Mice
Victoria P. Le 1, Attila Kovacs 2, Jessica E. Wagenseil 1
1Department of Biomedical Engineering, Saint Louis University, 2Department of Internal Medicine, Washington University School of Medicine

Measuring left ventricular pressure (LV) in embryonic and neonatal mice is described. Pressure is measured by inserting a needle connected to a fluid-filled transducer into the LV under ultrasound guidance. Care must be taken to maintain normal cardiac function during the experimental protocol.

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Immunology and Infection

Recurrent Herpetic Stromal Keratitis in Mice, a Model for Studying Human HSK
Jessica Morris 1, Patrick M. Stuart 1, Megan Rogge 1, Chloe Potter 1, Nipun Gupta 1, Xiao-Tang Yin 1
1Department of Ophthalmology, Saint Louis University

Most studies of herpetic corneal disease use a primary infection model. However, primary infection with HSV-1 does not typically lead to human disease. Here we describe a recurrent model of herpetic corneal disease, which more closely mimics human disease.

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JoVE Core

Scanning-probe Single-electron Capacitance Spectroscopy
Kathleen A. Walsh 1, Megan E. Romanowich 1, Morewell Gasseller 1,2, Irma Kuljanishvili 1,3, Raymond Ashoori 4, Stuart Tessmer 1
1Department of Physics and Astronomy, Michigan State University, 2Department of Chemistry & Biochemistry/Physics, Mercyhurst University, 3Department of Physics, Saint Louis University, 4Department of Physics, Massachusetts Institute of Technology

Scanning-probe single-electron capacitance spectroscopy facilitates the study of single-electron motion in localized subsurface regions. A sensitive charge-detection circuit is incorporated into a cryogenic scanning probe microscope to investigate small systems of dopant atoms beneath the surface of semiconductor samples.

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Medicine

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation
Xiao-Tang Yin 1, Deena A. Tajfirouz 1, Patrick M. Stuart 1
1Department of Ophthalmology, Saint Louis University

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

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Bioengineering

Simple Polyacrylamide-based Multiwell Stiffness Assay for the Study of Stiffness-dependent Cell Responses
Sana Syed 1, Amin Karadaghy 1, Silviya Zustiak 1
1Biomedical Engineering Department, Saint Louis University

Here, a method that enables quick, efficient, and inexpensive preparation of polyacrylamide gels in a multiwell plate format is described. The method does not require any specialized equipment and could be easily adopted by any research laboratory. It would be particularly useful in research focused on understanding stiffness-dependent cell responses.

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Biology

Validation of a Mouse Model to Disrupt LINC Complexes in a Cell-specific Manner
David Razafsky 1, Chloe Potter 1, Didier Hodzic 1
1Department of Ophthalmology and Visual Sciences, Washington University in St. Louis School of Medicine

Nuclear envelope proteins play a central role in many basic biological processes and have been implicated in a variety of human diseases. This protocol describes a new Cre/Lox-based mouse model that allows for the spatiotemporal control of LINC complexes disruption.

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Neuroscience

Whole-cell Currents Induced by Puff Application of GABA in Brain Slices
Yangjian Feng *1,2, Binliang Tang *1,2, Ming Chen 2, Li Yang 1,3,4,5
1School of Psychology, South China Normal University, 2School of Life Sciences, South China Normal University, 3Brain Science Institute, South China Normal University, 4Center for Studies of Psychological Application, South China Normal University, 5Guangdong Key Laboratory of Mental Health and Cognitive Science, South China Normal University

We describe the puff technique, by which pharmacological reagents can be administered during whole-cell patch-clamp recording, and highlight various aspects of the features that are crucial for its success.

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Biochemistry

Measuring Trans-Plasma Membrane Electron Transport by C2C12 Myotubes
Shannon C. Kelly *1, Amanda M. Eccardt *1, Jonathan S. Fisher 1
1Department of Biology, Saint Louis University

The goal of this protocol is to spectrophotometrically monitor trans-plasma membrane electron transport utilizing extracellular electron acceptors and to analyze enzymatic interactions that may occur with these extracellular electron acceptors.

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Environment

Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris
Katherine M. Martin 1, Elizabeth A. Hasenmueller 2, John R. White 3, Lisa G. Chambers 4, Jeremy L. Conkle 1
1Department of Physical and Environmental Sciences, Texas A&M University-Corpus Christi, 2Department of Earth and Atmospheric Sciences, Saint Louis University, 3Department of Oceanography and Coastal Sciences, Louisiana State University, 4Department of Biology, University of Central Florida

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

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Immunology and Infection

Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse
James D. Brien *1, Mariah Hassert *1, E. Taylor Stone 1, Elizabeth Geerling 1, Lillian Cruz-Orengo 2, Amelia K. Pinto 1
1Department of Molecular Microbiology and Immunology, Saint Louis University, 2School of Veterinary Medicine University of California Davis

The goal of the protocol is to demonstrate the techniques used to investigate viral disease by isolating and quantifying Zika virus, from multiple organs in a mouse following infection.

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Chemistry

Versatile Technique to Produce a Hierarchical Design in Nanoporous Gold
Palak Sondhi 1, Dharmendra Neupane 1, Jay K. Bhattarai 2, Hafsah Ali 1, Alexei V. Demchenko 3, Keith J. Stine 1
1Department of Chemistry and Biochemistry, University of Missouri-Saint Louis, 2Mallinckrodt Pharmaceuticals Company, 3Department of Chemistry, Saint Louis University

Nanoporous gold with a hierarchical and bimodal pore size distribution can be produced by combining electrochemical and chemical dealloying. The composition of the alloy can be monitored via EDS-SEM examination as the dealloying process advances. The material's loading capacity can be determined by studying protein adsorption onto the material.

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Bioengineering

Tumor Spheroid Fabrication and Encapsulation in Polyethylene Glycol Hydrogels for Studying Spheroid-Matrix Interactions
Joseph Bruns 1, Shabnam Nejat 1, Allison Faber 1, Silviya P. Zustiak 1
1Biomedical Engineering Department, Saint Louis University

Here, we present a protocol that enables fast, robust, and cheap fabrication of tumor spheroids followed by hydrogel encapsulation. It is widely applicable as it does not require specialized equipment. It would be particularly useful for exploring spheroid-matrix interactions and building in vitro tissue physiology or pathology models.

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