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Texas Children's Hospital

11 ARTICLES PUBLISHED IN JoVE

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Neuroscience

A Rapid Approach to High-Resolution Fluorescence Imaging in Semi-Thick Brain Slices
Jennifer Selever 1, Jian-Qiang Kong 2, Benjamin R. Arenkiel 3,4
1Department of Molecular & Human Genetics, Baylor College of Medicine (BCM), 2Precisionary Instruments Inc., 3Departments of Molecular & Human Genetics and Neuroscience, Baylor College of Medicine (BCM), 4Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital

Here we describe a rapid and simple method to image fluorescently labeled cells in semi-thick brain slices. By fixing, slicing, and optically clearing brain tissue we describe how standard epifluorescent or confocal imaging can be used to visualize individual cells and neuronal networks within intact nervous tissue.

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Neuroscience

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue
Kevin Ung 1, Benjamin R. Arenkiel 1,2,3
1Department of Molecular & Human Genetics, Baylor College of Medicine (BCM), 2Department of Neuroscience, Baylor College of Medicine (BCM), 3Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

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Medicine

Implantation of Total Artificial Heart in Congenital Heart Disease
Iki Adachi 1,2, David S. L. Morales 3
1Congenital Heart Surgery, Texas Children's Hospital, 2Michael E. DeBakey Department of Surgery, Baylor College of Medicine, 3Cincinnati Children's Hospital Medical Center, The University of Cincinnati College of Medicine

This is a case report of a patient with congenitally corrected transposition of the great arteries (CCTGA) who received a total artificial heart (TAH) as a bridge to heart transplant. The TAH was successfully implanted with modifications to accommodate the patient's congenitally malformed heart.

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Immunology and Infection

Super-resolution Imaging of the Natural Killer Cell Immunological Synapse on a Glass-supported Planar Lipid Bilayer
Peilin Zheng *1,2, Grant Bertolet *1,2,3, Yuhui Chen 1,2, Shengjian Huang 1,2, Dongfang Liu 1,2,3
1Center for Human Immunobiology, Texas Children's Hospital, 2Department of Pediatrics, Baylor College of Medicine, 3Department of Pathology and Immunology, Baylor College of Medicine

We describe here a combination of the glass-supported lipid bilayer technique of forming immunological synapses with the super-resolution imaging technique of stimulated emission depletion (STED) microscopy. The goal of this protocol is to provide users with the instructions necessary to successfully carry out these two techniques.

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JoVE Journal

An Array-based Comparative Genomic Hybridization Platform for Efficient Detection of Copy Number Variations in Fast Neutron-induced Medicago truncatula Mutants
Yuhui Chen 1, Xianfu Wang 2, Shunfei Lu 3, Hongcheng Wang 2, Shibo Li 2, Rujin Chen 1
1Laboratory of Plant Genetics and Development, Noble Research Institute, 2Genetics Laboratory, University of Oklahoma Health Science Center, 3Medicine and Health School, Li Shui University

This protocol provides experimental steps and information about reagents, equipment, and analysis tools for researchers who are interested in carrying out whole genome array-based comparative genomic hybridization (CGH) analysis of copy number variations in plants.

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Neuroscience

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice
Gary Liu *1,2, Jay M. Patel *2,3, Burak Tepe 1, Cynthia K. McClard 2,4, Jessica Swanson 4, Kathleen B. Quast 4, Benjamin R. Arenkiel 1,3,4,5
1Program in Developmental Biology, Baylor College of Medicine, 2Medical Scientist Training Program, Baylor College of Medicine, 3Department of Neuroscience, Baylor College of Medicine, 4Department of Molecular and Human Genetics, Baylor College of Medicine, 5Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

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JoVE Journal

Navigating MARRVEL, a Web-Based Tool that Integrates Human Genomics and Model Organism Genetics Information
Julia Wang 1,2, Zhandong Liu 3,4, Hugo J. Bellen 1,4,5,6,7, Shinya Yamamoto 1,4,5,6
1Program in Developmental Biology, Baylor College of Medicine, 2Medical Scientist Training Program, Baylor College of Medicine, 3Department of Pediatrics, Baylor College of Medicine, 4Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, 5Department of Molecular and Human Genetics, Baylor College of Medicine, 6Department of Neuroscience, Baylor College of Medicine, 7Howard Hughes Medical Institute, Baylor College of Medicine

Here, we present a protocol to access and analyze many human and model organism databases efficiently. This protocol demonstrates the use of MARRVEL to analyze candidate disease-causing variants identified from next-generation sequencing efforts.

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Genetics

In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
J. Michael Harnish *1, Samantha L. Deal *2, Hsiao-Tuan Chao 1,3,4,5, Michael F. Wangler 1,2,4, Shinya Yamamoto 1,2,4,5
1Department of Molecular and Human Genetics, Baylor College of Medicine, 2Program in Developmental Biology, Baylor College of Medicine, 3Department of Pediatrics, Section of Neurology and Developmental Neuroscience, Baylor College of Medicine, 4Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, 5Department of Neuroscience, Baylor College of Medicine

The goal of this protocol is to outline the design and performance of in vivo experiments in Drosophila melanogaster to assess the functional consequences of rare gene variants associated with human diseases.

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Cancer Research

Natural Killer (NK) and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line
Minh Ma 1,2, Saiaditya Badeti 1, James K. Kim 1, Dongfang Liu 1,3
1Department of Pathology, Immunology and Laboratory Medicine, Rutgers-New Jersey Medical School, 2Department of Microbiology, Biochemistry & Molecular Genetics, Public Health Research Institute Center, New Jersey Medical School, Rutgers University, 3Center for Immunity and Inflammation, New Jersey Medical School, Rutgers-The State University of New Jersey

Here, we present a method to expand peripheral blood natural killer (PBNK), NK cells from liver tissues, and chimeric antigen receptor (CAR)-NK cells derived from peripheral blood mononuclear cells (PBMCs) or cord blood (CB). This protocol demonstrates the expansion of NK and CAR-NK cells using 221-mIL-21 feeder cells in addition to the optimized purity of expanded NK cells.

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Neuroscience

Imaging and Quantification of Intact Neuronal Dendrites via CLARITY Tissue Clearing
Brandon T. Pekarek 1, Patrick J. Hunt 1,2, Benjamin D. W. Belfort 1,2, Gary Liu 2, Benjamin R. Arenkiel 1,3
1Department of Genetics and Genomics, Baylor College of Medicine, 2Medical Scientist Training Program, Baylor College of Medicine, 3Department of Neuroscience, Baylor College of Medicine

Neuronal dendritic morphology often underlies function. Indeed, many disease processes that affect the development of neurons manifest with a morphological phenotype. This protocol describes a simple and powerful method for analyzing intact dendritic arbors and their associated spines.

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Neuroscience

Molecular Imaging of Human Brain Organoids Using Mass Spectrometry
Saleh M. Khalil *1,2, Gerarda Cappuccio *1,2, Feng Li 3,4, Mirjana Maletic-Savatic 1,2,4,5
1Department of Pediatrics -Neurology, Baylor College of Medicine, 2Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, 3Department of Pathology & Immunology, Baylor College of Medicine, 4Center for Drug Discovery, Baylor College of Medicine, 5Department of Neuroscience, Baylor College of Medicine

An advanced method was developed for mass spectrometry imaging (MSI) of brain organoids that allows mapping metabolite distributions within these models. This technology offers insights into brain metabolic pathways and metabolite signatures during early development and in disease, promising a deeper understanding of the human brain function.

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