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Texas A&M Health Science Center

10 ARTICLES PUBLISHED IN JoVE

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Biology

Live Cell Response to Mechanical Stimulation Studied by Integrated Optical and Atomic Force Microscopy
Andreea Trache 1,2, Soon-Mi Lim 1
1Department of Systems Biology and Translational Medicine, College of Medicine, Cardiovascular Research Institute, Texas A&M Health Science Center, 2Department of Biomedical Engineering, Texas A&M University

This paper aims to instruct the reader in the operation of an integrated atomic force-optical imaging microscope for mechanical stimulation of live cells in culture. A step-by-step protocol is presented. A representative data set that shows live cell response to mechanical stimulation is presented.

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Immunology and Infection

Using Luciferase to Image Bacterial Infections in Mice
Mi Hee Chang 1, Suat L.G. Cirillo 1, Jeffrey D. Cirillo 1
1Microbial & Molecular Pathogenesis, Texas A&M Health Science Center

Methods for bioluminescence imaging of bacterial infections in living animals are decribed. Pathogens are modified to express luciferase allowing optical whole body imaging of infections in live animals. Animal models can be infected with luciferase expressing pathogens and the resulting course of disease visualized in real-time by bioluminescence imaging.

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Bioengineering

Polymer Microarrays for High Throughput Discovery of Biomaterials
Andrew L. Hook 1, Chien-Yi Chang 2, Jing Yang 1, David J. Scurr 1, Robert Langer 3, Daniel G. Anderson 3, Steve Atkinson 2, Paul Williams 2, Martyn C. Davies 1, Morgan R. Alexander 1
1Laboratory of Biophysics and Surface Analysis, University of Nottingham , 2School of Molecular Medical Sciences, University of Nottingham , 3David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology

A description of the formation of a polymer microarray using an on-chip photopolymerization technique. The high throughput surface characterization using atomic force microscopy, water contact angle measurements, X-ray photoelectron spectroscopy and time of flight secondary ion mass spectrometry and a cell attachment assay is also described.

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JoVE Journal

Mouse Fetal Liver Culture System to Dissect Target Gene Functions at the Early and Late Stages of Terminal Erythropoiesis
Baobing Zhao 1, Yang Mei 1, Jing Yang 1, Peng Ji 1
1Department of Pathology, Northwestern University

We present an in vitro mouse fetal liver erythroblast culture system that dissects the early and late stages of terminal erythropoiesis. This system facilitates functional analysis of specific genes in different developmental stages.

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Medicine

A Simple Critical-sized Femoral Defect Model in Mice
Bret H. Clough 1, Matthew R. McCarley 2, Carl A. Gregory 1,3
1Institute for Regenerative Medicine at Scott & White Hospital, Texas A&M Health Science Center, 2Department of Orthopedic Surgery, University of Texas Medical Branch, 3Molecular and Cellular Medicine, Texas A&M Health Science Center

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

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JoVE Journal

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
Vishnu V. Krishnamurthy *1, Aurora J. Turgeon *2, John S. Khamo 1, Payel Mondal 1, Savanna R. Sharum 1, Wenyan Mei 2, Jing Yang 2, Kai Zhang 1,3,4
1Department of Biochemistry, University of Illinois at Urbana-Champaign, 2Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 3Neuroscience Program, University of Illinois at Urbana-Champaign, 4Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign

This protocol describes an optogenetic strategy to modulate mitogen-activated protein kinase (MAPK) activity during cell differentiation and Xenopus embryonic development. This method allows for the reversible activation of the MAPK signaling pathway in mammalian cell culture and in multicellular live organisms, like Xenopus embryos, with high spatial and temporal resolution.

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Immunology and Infection

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence
Riti Sharan *1, Hee-Jeong Yang *2, Preeti Sule 1, Jeffrey D. Cirillo 1
1Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, 2Tuberculosis Research Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health

We describe the optical imaging of mice infected with Mycobacterium tuberculosis (M. tuberculosis) using reporter enzyme fluorescence (REF). This protocol facilitates the sensitive and specific detection of M. tuberculosis in pre-clinical animal models for pathogenesis, therapeutics and vaccine research.

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Bioengineering

Fabrication and Characterization of Optical Tissue Phantoms Containing Macrostructure
Madeleine S. Durkee 1, Landon D. Nash 1, Fatemeh Nooshabadi 1, Jeffrey D. Cirillo 2, Duncan J. Maitland 1, Kristen C. Maitland 1
1Department of Biomedical Engineering, Texas A&M University, 2Deparment of Molecular Pathogenesis and Immunology, Texas A&M College of Medicine

Optical tissue phantoms are essential tools for calibration and characterization of optical imaging systems and validation of theoretical models. This article details a method for phantom fabrication that includes replication of tissue optical properties and three-dimensional tissue structure.

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Developmental Biology

A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos
Sanat Upadhyay 1, Leoncio Vergara 2, Pranjali Shah 2, Jan-Åke Gustafsson 3,4, Ioannis Kakadiaris 1,4, Maria Bondesson 5
1Computational Biomedicine Lab, Texas Institute of Measurement Evaluation and Statistics, University of Houston, 2Institute of Biosciences and Technology, Texas A&M Health Science Center, 3Department of Biology and Biochemistry, Center for Nuclear Receptors and Cell Signaling, University of Houston, 4Department of Biosciences and Nutrition, Novum, Karolinska Institutet, 5Department of Intelligent Systems Engineering, Indiana University

This article describes a method to mount fragile zebrafish embryos for extended time-lapse confocal microscopy. This low-cost method is easy to perform using regular glass-bottom microscopy dishes for imaging on any inverted microscope. The mounting is performed in layers of agarose at different concentrations.

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Immunology and Infection

Automated, High-Throughput Detection of Bacterial Adherence to Host Cells
Jing Yang 1, Qing-Ming Qin 1, Erin Van Schaik 1, James E. Samuel 1, Paul de Figueiredo 1,2
1Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, 2College of Veterinary Medicine, Texas A&M University

Detection of host-bacterial pathogen interactions based on phenotypic adherence using high-throughput fluorescence labeling imaging along with automated statistical analysis methods enables rapid evaluation of potential bacterial interactions with host cells.

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