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Washington University in St. Louis

39 ARTICLES PUBLISHED IN JoVE

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Biology

Pyrosequencing: A Simple Method for Accurate Genotyping
Cristi King 1, Tiffany Scott-Horton 1
1Department of Internal Medicine, Washington University in St. Louis

Pyrosequencing(R) is one of the most thorough yet simple methods to date used to analyze polymorphisms. This method has led to rapid and efficient single-nucleotide polymorphism evaluation including many clinically relevant polymorphisms. The technique and methodology of Pyrosequencing is explained.

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Biology

Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System
Amos Danielli 1,2, Noga Porat 3, Marcelo Ehrlich 4, Ady Arie 1
1Department of Physical Electronics, Faculty of Engineering, Tel Aviv University, 2Department of Biomedical Engineering, Washington University in St. Louis, 3Department of Biological Sciences, University of Illinois, 4Department of Cell Research and Immunology, Tel Aviv University

Magnetic modulation biosensing system is utilized to rapidly, sensitively and simply detect biological assays, such as DNA molecules and proteins.

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Immunology and Infection

Non-invasive Imaging of Leukocyte Homing and Migration in vivo
Baomei Wang 1, Bernd H. Zinselmeyer 1,2, Jeremiah R. McDole 1, Peggy A. Gieselman 1, Mark J. Miller 1
1Department of Pathology and Immunology, Washington University in St. Louis, 2National Institute of Neurological Disorders and Stroke, NINDS, NIH - National Institute of Health

Here, we describe a non-invasive two-photon (2P) microscopy approach to study leukocyte homing in the mouse footpad. We discuss the technical aspects of our tissue imaging preparation and walk the reader through a typical experiment from initial set up to execution and data collection.

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Biology

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes
Junqiu Yang 1, Kelli Delaloye 2, Urvi S. Lee 2, Jianmin Cui 3
1Department of Energy, Environmental & Chemical Engineering, Washington University in St. Louis, 2Department of Biomedical Engineering, Washington University in St. Louis, 3Department of Biomedical Engineering and Cardiac Bioelectricity and Arrhythmia Center, Washington University in St. Louis

Ionic current of BK channels is recorded using patch clamp techniques. BK channels are expressed in Xenopus oocytes by injecting messenger RNA. The intracellular solution during patch clamp recordings is controlled by a perfusion system.

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Bioengineering

Three-dimensional Optical-resolution Photoacoustic Microscopy
Song Hu 1, Konstantin Maslov 1, Lihong V. Wang 1
1Optical Imaging Laboratory, Department of Biomedical Engineering, Washington University in St. Louis

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

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Bioengineering

Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart
Qing Lou 1, Wenwen Li 1, Igor R. Efimov 1
1Department of Biomedical Engineering, Washington University in St. Louis

This article describes the basic procedures for conducting optical mapping experiments in the Langendorff-perfused rabbit heart using the panoramic imaging system, and the dual (voltage and calcium) imaging modality.

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Bioengineering

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement
Dawn M. Johnson 1, Natalie A. LaFranzo 1, Joshua A. Maurer 1
1Department of Chemistry, Washington University in St. Louis

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

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Bioengineering

Optical Mapping of Action Potentials and Calcium Transients in the Mouse Heart
Di Lang 1, Matthew Sulkin 1, Qing Lou 1, Igor R. Efimov 1
1Department of Biomedical Engineering, Washington University in St. Louis

This paper details the dissection procedure, instrumental setup, and experimental conditions during optical mapping of transmembrane potential (Vm) and intracellular calcium transient (CaT) in intact isolated Langendorff perfused mouse hearts.

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Engineering

Quantitative Strain Mapping of an Abdominal Aortic Aneurysm
Hannah L. Cebull 1, Arvin H. Soepriatna 1, John J. Boyle 2, Craig J. Goergen 1
1Weldon School of Biomedical Engineering, Purdue University, 2Mechanical Engineering & Materials Science, Washington University in St. Louis

Quantitative Strain Mapping of an Abdominal Aortic Aneurysm

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Environment

Optimize Flue Gas Settings to Promote Microalgae Growth in Photobioreactors via Computer Simulations
Lian He 1, Amelia B Chen 1,2, Yi Yu 2, Leah Kucera 3, Yinjie Tang 1
1Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, 2School of Pharmaceutical Sciences, Wuhan University of China, 3Department of Earth and Planetary Sciences, Washington University in St. Louis

Flue gas from power plants is a cheap CO2 source for algal growth. We have built prototype "flue gas to algal cultivation" systems and described how to scale up the algal cultivation process. We have demonstrated the use of a mass-transfer bio-reaction model to simulate and to design the optimal operation of flue gas for the growth of Chlorella sp. in algal photo-bioreactors.

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Biology

The Xenopus Oocyte Cut-open Vaseline Gap Voltage-clamp Technique With Fluorometry
Michael W. Rudokas 1, Zoltan Varga 1, Angela R. Schubert 1, Alexandra B. Asaro 1, Jonathan R. Silva 1
1Department of Biomedical Engineering, Washington University in St. Louis

The cut-open Vaseline gap approach is used to obtain low noise recordings of ionic and gating currents from voltage-dependent ion channels expressed in Xenopus oocytes with high resolution of fast channel kinetics. With minor modification, voltage clamp fluorometry can be coupled to the cut-open oocyte protocol.

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Bioengineering

Quantification of Global Diastolic Function by Kinematic Modeling-based Analysis of Transmitral Flow via the Parametrized Diastolic Filling Formalism
Sina Mossahebi 2,5, Simeng Zhu 2,5, Howard Chen 1,5, Leonid Shmuylovich 3,5, Erina Ghosh 1,5, Sándor J. Kovács 4,5
1Department of Biomedical Engineering, Washington University in St. Louis, 2Department of Physics, Washington University in St. Louis, 3Division of Biology and Biomedical Sciences, Washington University in St. Louis, 4Department of Medicine, Cardiovascular Division, Washington University in St. Louis, 5Cardiovascular Biophysics Lab, Washington University in St. Louis

Accurate, causality-based quantification of global diastolic function has been achieved by kinematic modeling-based analysis of transmitral flow via the Parametrized Diastolic Filling (PDF) formalism. PDF generates unique stiffness, relaxation, and load parameters and elucidates 'new' physiology while providing sensitive and specific indexes of dysfunction.

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Neuroscience

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb
Wayne I. Doyle 1, Gary F. Hammen 2, Julian P. Meeks 1
1Department of Neuroscience, UT Southwestern Medical Center, 2Department of Anatomy and Neurobiology, Washington University in St. Louis

The mouse accessory olfactory bulb (AOB) has been difficult to study in the context of sensory coding. Here, we demonstrate a dissection that produces an ex vivo preparation in which AOB neurons remain functionally connected to their peripheral inputs, facilitating research into information processing of mouse pheromones and kairomones.

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Medicine

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo
Cara C. Rada 1, Stephanie L. Pierce 2, Chad A. Grotegut 2, Sarah K. England 1
1Department of Obstetrics and Gynecology, Washington University in St. Louis, 2Department of Obstetrics and Gynecology, Duke University

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

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Neuroscience

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice
Daniel S. Brenner 1,2, Judith P. Golden 2, Sherri K. Vogt 2, Robert W. Gereau IV 2
1MSTP, Neuroscience Program, Washington University in St. Louis, 2Washington University Pain Center, Department of Anesthesiology, Washington University in St. Louis

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 °C and 5 °C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

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Neuroscience

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System
Frans Vinberg 1, Vladimir Kefalov 1
1Department of Ophthalmology and Visual Sciences, Washington University in St. Louis

Ex vivo ERG can be used to record electrical activity of retinal cells directly from isolated intact retinas of animals or humans. We demonstrate here how common in vivo ERG systems can be adapted for ex vivo ERG recordings in order to dissect the electrical activity of retinal cells.

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Neuroscience

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera
Eirik Søvik *1,2, Jenny A. Plath *3,4, Jean-Marc Devaud 5, Andrew B. Barron 3
1Department of Science and Mathematics, Volda University College, 2Department of Biology, Washington University in St. Louis, 3Department of Biological Sciences, Macquarie University, 4Department of Biology, University of Konstanz, 5Research Center on Animal Cognition, CNRS, Universite de Toulouse

This manuscript describes several protocols for administering pharmacological agents to honey bees, including simple noninvasive methods for free-flying bees, as well as more invasive variants that allow precise localized treatment of restrained bees.

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Biology

High-resolution Optical Mapping of the Mouse Sino-atrial Node
Di Lang 1, Alexey V. Glukhov 1
1Department of Medicine, University of Wisconsin-Madison School of Medicine and Public Health

Here, we present a protocol for optical mapping of electrical activity from the mouse right atrium and especially the sino-atrial node, at a high spatial and temporal resolution.

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Biology

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)
Jing Men 1,2, Jason Jerwick 2,3, Penghe Wu 1,2, Mingming Chen 3,4, Aneesh Alex 2,3, Yutao Ma 4, Rudolph E. Tanzi 5, Airong Li 5, Chao Zhou 1,2,3
1Bioengineering Program, Lehigh University, 2Center for Photonics and Nanoelectronics, Lehigh University, 3Department of Electrical and Computer Engineering, Lehigh University, 4State Key Laboratory of Software Engineering, Wuhan University, 5Genetics and Aging Research Unit, Department of Neurology, Massachusetts General Hospital and Harvard Medical School

Here, the experimental protocols are described for preparing Drosophila at different developmental stages and performing longitudinal optical imaging of Drosophila heartbeats using a custom optical coherence microscopy (OCM) system. The cardiac morphological and dynamical changes can be quantitatively characterized by analyzing the heart structural and functional parameters from OCM images.

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Developmental Biology

Directed Differentiation of Primitive and Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells
Carissa Dege 1, Christopher M. Sturgeon 1
1Department of Internal Medicine, Hematology Division, Washington University in St. Louis

Here, we present human pluripotent stem cell (hPSC) culture protocols, used to differentiate hPSCs into CD34+ hematopoietic progenitors. This method uses stage-specific manipulation of canonical WNT signaling to specify cells exclusively to either the definitive or primitive hematopoietic program.

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Bioengineering

An In Vitro Organ Culture Model of the Murine Intervertebral Disc
Jennifer W. Liu 1,3, Kevin H. Lin 2, Christian Weber 1, Sameer Bhalla 2, Sean Kelso 4, Kaixi Wang 3, Simon Y. Tang 1,3,4
1Department of Biomedical Engineering, Washington University in St. Louis, 2Department of Biology, Washington University in St. Louis, 3Department of Orthopaedic Surgery, Washington University in St. Louis, 4Department of Materials Science and Mechanical Engineering, Washington University in St. Louis

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

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Education

Photoelectron Imaging of Anions Illustrated by 310 Nm Detachment of F
Justin Lyle 1, Sudharson Ravishankar Chandramoulee 1, C. Annie Hart 1, Richard Mabbs 1
1Department of Chemistry, Washington University in St. Louis

Here, we present a protocol for photoelectron imaging of anionic species. Anions generated in vacuo and separated by mass spectrometry are probed using velocity mapped photoelectron imaging, providing details of anion and neutral energy levels, anion and neutral structure and the nature of the anion electronic state.

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Cancer Research

Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography
Yongyang Huang 1, Jinyun Zou 1, Mudabbir Badar 1, Junchao Liu 1, Wentao Shi 5, Shunqiang Wang 2, Qiongyu Guo 3, Xiaofang Wang 1, Sarah Kessel 4, Leo Li-Ying Chan 4, Peter Li 4, Yaling Liu 2,5, Jean Qiu 4, Chao Zhou 1,5,6
1Department of Electrical and Computer Engineering, Lehigh University, 2Department of Mechanical Engineering, Lehigh University, 3Department of Biomedical Engineering, Southern University of Science and Technology, 4Department of Technology R&D, Nexcelom Bioscience LLC, 5Department of Bioengineering, Lehigh University, 6Center for Photonics and Nanoelectronics, Lehigh University

Optical coherence tomography (OCT), a three-dimensional imaging technology, was used to monitor and characterize the growth kinetics of multicellular tumor spheroids. Precise volumetric quantification of tumor spheroids using a voxel counting approach, and label-free dead tissue detection in the spheroids based on intrinsic optical attenuation contrast, were demonstrated.

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Chemistry

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
Brian J. Stockman 1, Abinash Kaur 1, Julia K. Persaud 1, Maham Mahmood 1, Samantha F. Thuilot 1, Melissa B. Emilcar 1, Madison Canestrari 1, Juliana A. Gonzalez 1, Shannon Auletta 1, Vital Sapojnikov 1, Wagma Caravan 1,2, Samantha N. Muellers 1,3
1Department of Chemistry, Adelphi University, 2Department of Chemistry, Washington University in St. Louis, 3Department of Chemistry, Boston University

NMR-based activity assays have been developed to identify and characterize inhibitors of two nucleoside ribohydrolase enzymes. Protocols are provided for initial compound assays at 500 μM and 250 μM, dose-response assays for determining IC50 values, detergent counter screen assays, jump-dilution counter screen assays, and assays in E. coli whole cells.

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Bioengineering

Preclinical Cardiac Electrophysiology Assessment by Dual Voltage and Calcium Optical Mapping of Human Organotypic Cardiac Slices
Sharon A. George *1, Jaclyn A. Brennan *1, Igor R. Efimov 1
1Department of Biomedical Engineering, The George Washington University

This protocol describes the procedure for sectioning and culturing human cardiac slices for preclinical drug testing and details the use of optical mapping for recording transmembrane voltage and intracellular calcium signals simultaneously from these slices.

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Neuroscience

Primary Microglia Isolation from Postnatal Mouse Brains
Siling Du *1,2, Shanshan Xiong *3, Xiangjuan Du 4, Ti-Fei Yuan 1, Bo Peng 3,4,5, Yanxia Rao 1
1Shanghai Key Laboratory of Psychotic Disorders, Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine, 2Division of Biology and Biomedical Science, Washington University in St. Louis, 3Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 4Department of Neurosurgery, Huashan Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, MOE Frontiers Center for Brain Science, Fudan University, 5Co-innovation Center of Neuroregeneration, Nantong University

Primary cell culture is one of the primarily used approaches for studying microglial biology in vitro. Here, we developed a method for simple and rapid microglia isolation from the mouse postnatal day 1 (P1) to P4.

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Chemistry

Comparison of Two Different Synthesis Methods of Single Crystals of Superconducting Uranium Ditelluride
Sheng Ran 1,2,3, I-Lin Liu 1,2, Shanta R. Saha 1,2, Prathum Saraf 1, Johnpierre Paglione 1,2, Nicholas P. Butch 1,2
1Maryland Quantum Materials Center, Department of Physics, University of Maryland, 2National Institute of Standards and Technology, 3Department of Physics, Washington University in St. Louis

Here, we present a protocol to synthesize two types of UTe2 crystals: those exhibiting robust superconductivity, via chemical vapor transport synthesis, and those lacking superconductivity, via molten metal flux synthesis.

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Neuroscience

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings
Kristen N. Reikersdorfer *1, Andrea K. Stacy *2, David A. Bressler 2, Lauren S. Hayashi 2, Keith B. Hengen 1, Stephen D. Van Hooser 2
1Department of Biology, Washington University in St. Louis, 2Department of Biology, Program in Neuroscience, Brandeis University

This protocol describes a procedure for constructing carbon fiber microelectrode arrays for chronic and acute in vivo electrophysiological recordings in mouse (Mus musculus) and ferret (Mustela putorius furo) from multiple brain regions. Each step, following the purchase of raw carbon fibers to microelectrode array implantation, is described in detail, with emphasis on microelectrode array construction.

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Neuroscience

Optimizing the Setup and Conditions for Ex Vivo Electroretinogram to Study Retina Function in Small and Large Eyes
Fatima Abbas 1, Frans Vinberg 1, Silke Becker 1
1John A. Moran Eye Center, University of Utah

Modification of existing multielectrode array or patch clamp equipment makes the ex vivo electroretinogram more widely accessible. Improved methods to record and maintain ex vivo light responses facilitate the study of photoreceptor and ON-bipolar cell function in the healthy retina, animal models of eye diseases, and human donor retinas.

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Biology

Isolation of Cardiac and Vascular Smooth Muscle Cells from Adult, Juvenile, Larval and Embryonic Zebrafish for Electrophysiological Studies
Soma S. Singareddy 1, Conor McClenaghan 1, Helen I. Roessler 2, Robert Tryon 1, Colin G. Nichols 1
1Department of Cell Biology and Physiology and Center for the Investigation of Membrane Excitability Diseases, Washington University in St. Louis, 2Department of Genetics, Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University

The present protocol describes the acute isolation of viable cardiac and vascular smooth muscle cells from adult, juvenile, larval, and embryonic zebrafish (Danio rerio), suitable for electrophysiological studies.

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Genetics

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay
Davi J. Martins *1, Stephanie Tirman *2, Annabel Quinet 2, Carlos F. M. Menck 1
1Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, 2Division of Oncology, Department of Internal Medicine, Washington University in St. Louis

Here we describe two modifications of the DNA fiber assay to investigate single-stranded DNA gaps in replicating DNA after lesion induction. The S1 fiber assay enables the detection of post-replicative gaps using the ssDNA-specific S1 endonuclease, while the gap-filling assay allows visualization and quantification of gap repair.

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Bioengineering

Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function
Elena Gracheva 1, Fei Wang 1, Abigail Matt 1, Hongwu Liang 1, Matthew Fishman 1,2, Chao Zhou 1
1Department of Biomedical Engineering, Washington University in St. Louis, 2Department of Computer Science and Engineering, Washington University in St. Louis

The present protocol describes the generation of Drosophila melanogaster expressing eNpHR2.0 or ReaChR opsins in the heart for OCT imaging and optogenetic heart pacing. Detailed instructions for Drosophila OCT imaging and heart beating modulation, including the simulation of restorable heart arrest, bradycardia, and tachycardia in live animals at different developmental stages, are reported.

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Cancer Research

Generation and Culturing of High-Grade Serous Ovarian Cancer Patient-Derived Organoids
Olivia Graham *1, Jeimmy Rodriguez *1, Lillian van Biljon 1, Bisiayo Fashemi 1, Emily Graham 1, Katherine Fuh 1,2, Dineo Khabele 1, Mary Mullen 1
1Washington University in St. Louis, 2University of California San Francisco

Patient-derived organoids (PDO) are a three-dimensional (3D) culture that can mimic the tumor environment in vitro. In high-grade serous ovarian cancer, PDOs represent a model to study novel biomarkers and therapeutics.

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Cancer Research

Visualizing DNA Damage Repair Proteins in Patient-Derived Ovarian Cancer Organoids via Immunofluorescence Assays
Lillian van Biljon 1, Bisiayo Fashemi 1, Jeimmy Rodriguez 1, Olivia Graham 1, Amanda Compadre 1, Katherine Fuh 1,2, Dineo Khabele 1, Mary Mullen 1
1Washington University in St. Louis, 2University of California San Francisco

The present protocol describes methods for evaluating DNA damage repair proteins in patient-derived ovarian cancer organoids. Included here are comprehensive plating and staining methods, as well as detailed, objective quantification procedures.

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Biology

Decellularization-Based Quantification of Skeletal Muscle Fatty Infiltration
Jacob C. Parson 1, Nicole K. Biltz 1, Gretchen A. Meyer 1,2
1Program in Physical Therapy, Washington University in St. Louis, 2Departments of Neurology, Orthopaedic Surgery and Biomedical Engineering, Washington University in St. Louis

The present study describes decellularization-based methodologies for visualizing and quantifying intramuscular adipose tissue (IMAT) deposition through intact muscle volume, as well as quantifying metrics of individual adipocytes that comprise IMAT.

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Medicine

A Simple Cuff Technique for Murine Left Lung Transplantation
Zhongcheng Mei 1, Mojtaba Taheri 1, Elizabeth A. Jacobsen 2, Wenjun Li 3,4, Andrew E. Gelman 3,4, Daniel Kreisel 3,4, Alexander Sasha Krupnick 1,5
1Department of Surgery, University of Maryland Medical Center, 2Division of Allergy, Asthma and Clinical Immunology, Mayo Clinic, 3Department of Surgery, Washington University in St. Louis, 4Department of Pathology & Immunology, Washington University in St. Louis, 5Department of Microbiology and Immunology, University of Maryland Medical Center

The present protocol describes a cuff technique for a mouse left lung transplantation model. This technique has been developed over several years and has performed well, serving effectively in immunological research.

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Genetics

Concurrent Collection of Fetal Murine Brain and Serum to Assess Effects of Maternal Diet on Nutrition and Neurodevelopment in Neurofibromatosis Type 1
Gemma E. Martin 1, Ambrose Chan 1, Nicole M. Brossier 1
1Department of Pediatrics, Washington University in St. Louis

In this protocol, we describe a method for simultaneous collection of fetal brain tissue as well as high-quality, non-hemolyzed serum from the same mouse embryo. We have utilized this technique to interrogate how maternal dietary exposure affects macronutrient profiles and fetal neurodevelopment in mice heterozygous for Nf1 (Neurofibromatosis Type 1).

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Medicine

Intravital Two-Photon Microscopy of the Transplanted Mouse Lung
Yun Zhu Bai 1, Yuriko Terada 1, Katsutaka Mineura 1, Yuhei Yokoyama 1, Ruben G. Nava 1, Alexander S. Krupnick 2, Andrew E. Gelman 1, Mark J. Miller 3, Daniel Kreisel 1, Wenjun Li 1
1Division of Cardiothoracic Surgery, Washington University in St. Louis, 2Division of Thoracic Surgery, University of Maryland, 3Division of Infectious Diseases, Washington University in St. Louis

Here, we present a protocol to intravitally image the transplanted mouse left lung using two-photon microscopy. This represents a valuable tool for studying cellular dynamics and interactions in real-time following murine lung transplantation.

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Medicine

Determination of Continuity Index Values in Atrial Fibrillation Ablation with Proactive Esophageal Cooling
Catherine Lazarus 1, Jacob Sherman 2, Natalie Putzel 3, Tiffany Sharkoski 4, William Zagrodzky 4, Erik Kulstad 4,5, Alex Ro 6, Jose Nazari 6, Westby Fisher 6, Mark Metzl 6
1Northwestern University, 2Washington University in St. Louis, 3University of Southern California, 4Attune Medical, 5University of Texas Southwestern Medical Center, 6NorthShore University Hospital

This protocol describes the methods used to determine the continuity index in patients undergoing pulmonary vein isolation procedures using radiofrequency ablation and demonstrates the differences in continuity index between ablation procedures using proactive esophageal cooling as compared to procedures using traditional luminal esophageal temperature monitoring.

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