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The Pennsylvania State University

19 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Preparation of Viral DNA from Nucleocapsids
Moriah L. Szpara 1, Yolanda R. Tafuri 1, L. W. Enquist 1
1Department of Molecular Biology, Princeton University

We describe the process of isolating high purity herpesvirus nucleocapsid DNA from infected cells. The final DNA captured from solution is of high concentration and purity, making it ideally suited for high-throughput sequencing, high fidelity PCR reactions, and transfections to produce new viral recombinants.

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Engineering

Encapsulation and Permeability Characteristics of Plasma Polymerized Hollow Particles
Anaram Shahravan 1, Themis Matsoukas 1
1Department of Chemical Engineering, The Pennsylvania State University

We have used plasma enhanced chemical vapor deposition to deposit thin films ranging from a few nm to several 100 nm on nano-sized particles of various materials. We subsequently etch the core material to produce hollow nanoshells whose permeability is controlled by the thickness of the shell. We characterize the permeability of these coatings to small solutes and demonstrate that these barriers can provide sustained release of the core material over several days.

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Bioengineering

Molecular Entanglement and Electrospinnability of Biopolymers
Lingyan Kong 1, Gregory R. Ziegler 1
1Department of Food Science, Pennsylvania State University

Electrospinning is a fascinating technique used to fabricate micro- to nano-scale fibers from a wide variety of materials. Molecular entanglement of the constituent polymers in the spinning dope is essential for successful electrospinning. We present a protocol for utilizing rheology to evaluate the electrospinnability of two biopolymers, starch and pullulan.

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Behavior

Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors
Colleen McSweeney 1, Yingwei Mao 1
1Department of Biology, The Pennsylvania State University

Stereotactic injection of lentiviruses expressing cDNAs or shRNAs can modulate gene expression in specific brain areas of mice. Here, we present a protocol to combine stereotactic injections with behavioral tasks, such as the Open Field Test (OFT) and the Forced Swim Test (FST).

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Developmental Biology

Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line
Mackenzie M. Shipley 1, Colleen A. Mangold 1, Moriah L. Szpara 1
1Department of Biochemistry and Molecular Biology, The Huck Institutes of the Life Sciences, The Pennsylvania State University

It is critical in neurobiology and neurovirology to have a reliable, replicable in vitro system that serves as a translational model for what occurs in vivo in human neurons. This protocol describes how to culture and differentiate SH-SY5Y human neuroblastoma cells into viable neurons for use in in vitro applications.

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Engineering

Characterization of Full Set Material Constants and Their Temperature Dependence for Piezoelectric Materials Using Resonant Ultrasound Spectroscopy
Liguo Tang 1, Wenwu Cao 2
1Key Laboratory of Underwater Acoustic Communication and Marine Information Technology, Xiamen University, 2Department of Mathematics and Material Research Institute, The Pennsylvania State University

This protocol describes the procedure of measuring the temperature dependence of the full set material constants of piezoelectric materials using resonant ultrasound spectroscopy (RUS).

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Chemistry

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology
Amir Sheikhi 1,2,3, Han Yang 1,2,3, Md. Nur Alam 1,2,3, Theo G. M. van de Ven 1,2,3
1Department of Chemistry, McGill University, 2Center for Self-Assembled Chemical Structures (CSACS), McGill University, 3Pulp and Paper Research Center, McGill University

Synthesis schemes to prepare highly stable wood fiber-based hairy nanoparticles and functional cellulose-based biopolymers have been detailed.

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Immunology and Infection

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Joselyn N. Allen 1, Adwitia Dey 1, Ruth Nissly 2, James Fraser 1, Shan Yu 1, Gayathri Balandaram 1, Jeffrey M. Peters 1, Pamela A. Hankey-Giblin 1
1Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, 2Microscopy and Cytometry Facility, The Huck Institutes of Life Sciences, Pennsylvania State University

This protocol allows a researcher to isolate and characterize tissue-resident macrophages in various hallmark inflamed tissues extracted from diet-induced models of metabolic disorders.

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Bioengineering

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions
Djoshkun Shengjuler 1, Simou Sun 2, Paul S. Cremer 1,2, Craig E. Cameron 1
1Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 2Department of Chemistry, The Pennsylvania State University

Here we present a supported lipid bilayer in the context of a microfluidic platform to study protein-phosphoinositide interactions using a label-free method based on pH modulation.

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Education

Determination of Thermodynamic Properties of Alkaline Earth-liquid Metal Alloys Using the Electromotive Force Technique
Thomas P. Nigl 1, Nathan D. Smith 1, Timothy Lichtenstein 1, Jarrod Gesualdi 1, Kuldeep Kumar 1, Hojong Kim 1
1Department of Materials Science and Engineering, The Pennsylvania State University

This protocol describes the measurement of the electromotive force of alkaline-earth elements in liquid metal alloys at high temperatures (723-1,123 K) to determine their thermodynamic properties, including activity, partial molar entropy, partial molar enthalpy, and phase transition temperatures, over a wide composition range.

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Behavior

Assessment of Midline Lingual Point-Pressure Somatosensation Using Von Frey Hair Monofilaments
Nicole M. Etter 1, Scott P. Breen 2,3, Maya I. M. Alcala 2,3, Gregory R. Ziegler 3, John E. Hayes 2,3
1Department of Communication Sciences and Disorders, College of Health and Human Development, The Pennsylvania State University, 2Sensory Evaluation Center, The Pennsylvania State University, 3Department of Food Science, College of Agricultural Sciences, The Pennsylvania State University

This work describes a standard method to assess tactile sensation at the midline of the tongue tip. Using Von Frey Hair (VFH) monofilaments, this protocol provides estimates of detection and discrimination threshold estimates for oral point pressure (OPP).

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Biology

Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis
Elena Dalla Benetta *1, Duverney Chaverra-Rodriguez *1, Jason L. Rasgon 2, Omar S. Akbari 1
1Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, San Diego, 2Department of Entomology, The Center for Infectious Disease Dynamics, The Huck Institutes of the Life Sciences, The Pennsylvania State University

Here, we describe methods for efficient pupal and adult injections in Nasonia vitripennis as accessible alternatives to embryo microinjection, enabling functional analysis of genes of interest using either RNA-silencing via RNA interference (RNAi) or gene knockout via CRISPR/Cas9 genome editing.

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Chemistry

Picometer-Precision Atomic Position Tracking through Electron Microscopy
Leixin Miao 1, Adrian Chmielewski 1, Debangshu Mukherjee 2, Nasim Alem 1
1Department of Materials Science & Engineering, The Pennsylvania State University, 2Center for Nanophase Materials Sciences, Oak Ridge National Laboratory

This work presents a workflow for atomic position tracking in atomic resolution transmission electron microscopy imaging. This workflow is performed using an open-source Matlab app (EASY-STEM).

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JoVE Core

The Role of Fabric in Frictional Properties of Phyllosilicate-Rich Tectonic Faults
Cristiano Collettini 1,4, Telemaco Tesei 2, Fabio Trippetta 1, Marco M. Scuderi 1, Eliza Richardson 3, Chris Marone 1,3, Giacomo Pozzi 4, Cecilia Viti 5
1Dipartimento di Scienze della Terra, Sapienza Università di Roma, 2Dipartimento di Geoscienze, Università di Padova, 3Department of Geoscience, The Pennsylvania State University, 4Istituto Nazionale di Geofisica e Vulcanologia, 5Dipartimento di Scienze Fisiche, della Terra e dell’Ambiente, Università di Siena

Friction of phyllosilicates-rich faults sheared in their in situ geometry is significantly lower than friction of their powdered equivalents.

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Biochemistry

Sample Preparation using a Lipid Monolayer Method for Electron Crystallographic Studies
Chloe D. Truong 1,2, Dewight R. Williams 3, Mary Zhu 1, Joseph Che-Yen Wang 4, Po-Lin Chiu 1,2
1School of Molecular Sciences, Arizona State University, 2Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 3Eyring Materials Center, Arizona State University, 4Department of Microbiology and Immunology, The Pennsylvania State University

Lipid monolayers have been used as a foundation for forming two-dimensional (2D) protein crystals for structural studies for decades. They are stable at the air-water interface and can serve as a thin supporting material for electron imaging. Here we present the proven steps on preparing lipid monolayers for biological studies.

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Bioengineering

Photodegradable Hydrogel Interfaces for Bacteria Screening, Selection, and Isolation
Niloufar Fattahi 1, Niloy Barua 1, André J. van der Vlies 2, Ryan R. Hansen 1
1Tim Taylor Department of Chemical Engineering, Kansas State University, 2Department of Materials Science and Engineering, The Pennsylvania State University

The use of photodegradable hydrogels to isolate bacterial cells by utilizing a high-resolution light pattering tool is reported. Essential experimental procedures, results, and advantages of the process are reviewed. The method enables rapid and inexpensive isolation of targeted bacteria showing rare or unique functions from heterogeneous communities or populations.

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Bioengineering

Gelatin Methacryloyl Granular Hydrogel Scaffolds: High-throughput Microgel Fabrication, Lyophilization, Chemical Assembly, and 3D Bioprinting
Zaman Ataie 1, Arian Jaberi 1, Sina Kheirabadi 1, Aneesh Risbud 2, Amir Sheikhi 1,2
1Department of Chemical Engineering, The Pennsylvania State University, 2Department of Biomedical Engineering, The Pennsylvania State University

This article describes protocols for high-throughput gelatin methacryloyl microgel fabrication using microfluidic devices, converting microgels to resuspendable powder (micro-aerogels), the chemical assembly of microgels to form granular hydrogel scaffolds, and developing granular hydrogel bioinks with preserved microporosity for 3D bioprinting.

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Cancer Research

Intra-Peritoneal Transplantation for Generating Acute Myeloid Leukemia in Mice
Fenghua Qian 1, Brooke E. Arner 1, Shaneice K. Nettleford 1, Robert F. Paulson 1, K. Sandeep Prabhu 1
1Department of Veterinary and Biomedical Sciences, Center for Molecular Immunology and Infectious Disease and Center for Molecular Toxicology and Carcinogenesis, Penn State Cancer Institute, The Pennsylvania State University

Here, intra-peritoneal injection of leukemia cells is utilized to establish and propagate acute myeloid leukemia (AML) in mice. This new method is effective in the serial transplantation of AML cells and can serve as an alternative for those who may experience difficulties and inconsistencies with intravenous injection in mice.

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Medicine

Intradermal Microdialysis: An Approach to Investigating Novel Mechanisms of Microvascular Dysfunction in Humans
Auni C. Williams *1, Virginia G. Content *1, Nathalie V. Kirby 1, Lacy M. Alexander 1
1Department of Kinesiology, The Pennsylvania State University

Intradermal microdialysis is a minimally invasive technique used to investigate microvascular function in health and disease. Both dose-response and local heating protocols can be utilized for this technique to explore mechanisms of vasodilation and vasoconstriction in the cutaneous circulation.

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