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Uniformed Services University

6 ARTICLES PUBLISHED IN JoVE

Neuroscience

Migratory Behavior of Cells Generated in Ganglionic Eminence Cultures

Marcin Gierdalski^*1, Thomas McFate^*1, Joseph Abbah², Sharon L. Juliano^1,2

¹Dept. of Anatomy, Physiology and Genetics, Uniformed Services University, ²Neuroscience Program, Uniformed Services University

Time lapse imaging of 3D tissue culture allows studying migratory behavior of individual cells originating from ganglionic eminence in reaction to fractionated protein extract from cerebral cortex.

Immunology and Infection

Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue

Tami T. Tamashiro¹, Clifton Lee Dalgard^1,2,3, Kimberly R. Byrnes²

¹Neuroscience Program, Uniformed Services University, ²Department of Anatomy, Physiology, and Genetics, Uniformed Services University, ³Molecular and Cell Biology, Uniformed Services University

Isolating primary microglia from the cellular heterogeneity of the brain is essential to investigate their role in both physiological and pathological conditions. This protocol describes a mechanical isolation and mixed cell culture technique that provides high yield and high purity, viable primary microglial cells for in vitro study and downstream applications.

Immunology and Infection

A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum

William G. Valiant¹, Joseph J. Mattapallil¹

¹Department of Microbiology and Immunology, Uniformed Services University

We describe a simple and easy protocol for measuring antibody dependent enhancement of infection by Zika virus convalescent serum using Dengue virus Reporter Viral Particles.

Developmental Biology

Controlled Cortical Impact Model of Mouse Brain Injury with Therapeutic Transplantation of Human Induced Pluripotent Stem Cell-Derived Neural Cells

Orion Furmanski^1,2, Michael D. Nieves^1,2,3, Martin L. Doughty^1,2,3

¹Center for Neuroscience and Regenerative Medicine, Uniformed Services University, ²Department of Anatomy, Physiology, and Genetics, Uniformed Services University, ³Graduate Program in Neuroscience, Uniformed Services University

This protocol demonstrates methodologies for a mouse model of open-skull traumatic brain injury and transplantation of cultured human induced pluripotent stem cell-derived cells into the injury site. Behavioral and histologic tests of outcomes from these procedures are also described in brief.

Developmental Biology

Visualizing the Effects of Oxidative Damage on Drosophila Egg Chambers using Live Imaging

Kelsey M. Sheard¹, Rachel T. Cox¹

¹Department of Biochemistry and Molecular Biology, Uniformed Services University

The objective of this protocol is to use live imaging to visualize the effects of oxidative damage on the localization and dynamics of subcellular structures in Drosophila ovaries.

Cancer Research

Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

Dave Mitchell^*1,2, Allison L. Hunt^*1,4, Kelly A. Conrads^1,2, Brian L. Hood^1,2, Sasha C. Makohon-Moore^1,2, Christine Rojas¹, G. Larry Maxwell^1,3,4, Nicholas W. Bateman^1,2,3, Thomas P. Conrads^1,3,4

¹Women’s Health Integrated Research Center, Gynecologic Cancer Center of Excellence, Department of Obstetrics and Gynecology, Uniformed Services University and Walter Reed National Military Medical Center, ²The Henry M. Jackson Foundation for the Advancement of Military Medicine, ³The John P. Murtha Cancer Center Research Program, Department of Surgery, Uniformed Services University, ⁴Women’s Health Integrated Research Center, Inova Women’s Service Line, Inova Health System

This protocol describes a high-throughput workflow for artificial intelligence-driven segmentation of pathology-confirmed regions of interest from stained, thin tissue section images for enrichment of histology-resolved cell populations using laser microdissection. This strategy includes a novel algorithm enabling the transfer of demarcations denoting cell populations of interest directly to laser microscopes.