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University of Maine

9 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Non-invasive Imaging of Disseminated Candidiasis in Zebrafish Larvae
Kimberly M. Brothers 1, Robert T. Wheeler 1
1Department of Molecular and Biomedical Sciences, University of Maine

The rapid development, small size and transparency of zebrafish are tremendous advantages for the study of innate immune control of infection1-4. Here we demonstrate techniques for infecting zebrafish larvae using the fungal pathogen Candida albicans by microinjection, methodology recently used to implicate phagocyte NADPH oxidase activity in control of fungal dimorphism5.

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Immunology and Infection

Quantification of the Respiratory Burst Response as an Indicator of Innate Immune Health in Zebrafish
Michelle F. Goody 1, Eric Peterman 1, Con Sullivan 1, Carol H. Kim 1
1Department of Molecular and Biomedical Sciences, University of Maine

The innate immune response protects organisms against pathogen infection. A critical component of the innate immune response, the phagocyte respiratory burst, generates reactive oxygen species that kill invading microorganisms. We describe a respiratory burst assay that quantifies reactive oxygen species produced when the innate immune response is chemically induced.

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Biology

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
Nikki M. Curthoys *1, Michael J. Mlodzianoski *1, Dahan Kim 1, Samuel T. Hess 1
1Department of Physics and Astronomy, University of Maine

We demonstrate the use of fluorescence photo activation localization microscopy (FPALM) to simultaneously image multiple types of fluorescently labeled molecules within cells. The techniques described yield the localization of thousands to hundreds of thousands of individual fluorescent labeled proteins, with a precision of tens of nanometers within single cells.

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Immunology and Infection

Modeling Mucosal Candidiasis in Larval Zebrafish by Swimbladder Injection
Remi L. Gratacap 1, Audrey C. Bergeron 1, Robert T. Wheeler 1,2
1Department of Molecular and Biomedical Sciences, University of Maine, 2Graduate School of Biomedical Sciences and Engineering, University of Maine

In vivo spatio-temporal interactions of pathogen and immune defenses at the mucosal level are not easily imaged in existing vertebrate hosts. The method presented here describes a versatile platform to study mucosal candidiasis in live vertebrates using the swimbladder of the juvenile zebrafish as an infection site.

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Environment

Methods of Soil Resampling to Monitor Changes in the Chemical Concentrations of Forest Soils
Gregory B. Lawrence 1, Ivan J. Fernandez 2, Paul W. Hazlett 3, Scott W. Bailey 4, Donald S. Ross 5, Thomas R. Villars 6, Angelica Quintana 7, Rock Ouimet 8, Michael R. McHale 1, Chris E. Johnson 9, Russell D. Briggs 10, Robert A. Colter 11, Jason Siemion 1, Olivia L. Bartlett 12, Olga Vargas 13, Michael R. Antidormi 1, Mary M. Koppers 9
1New York Water Science Center, U.S. Geological Survey, 2School of Forest Resources, University of Maine, 3Natural Resources Canada, Canadian Forest Service, 4Northern Research Station, U.S. Forest Service, 5Department of Plant and Soil Science, University of Vermont, 6Ottauquechee NRCD, USDA Natural Resources Conservation Service, 7Green Mountain National Forest, U.S. Forest Service, 8Direction de la Recherche Forestière, Ministère du Québec, 9Department of Civil and Environmental Engineering, Syracuse University, 10Division of Environmental Science, SUNY College of Environmental Science and Forestry, 11White Mountain National Forest, U.S. Forest Service, 12Natural Resources and Earth System Sciences, University of New Hampshire, 13Greenwich, NY Field Office, USDA Natural Resources Conservation Service

Repeated soil sampling has recently been shown to be an effective way to monitor forest soil change over years and decades. To support its use, a protocol is presented that synthesizes the latest information on soil resampling methods to aid in the design and implementation of successful soil monitoring programs.

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Immunology and Infection

Using Zebrafish Models of Human Influenza A Virus Infections to Screen Antiviral Drugs and Characterize Host Immune Cell Responses
Con Sullivan 1,2, Denise Jurcyzszak 1, Michelle F. Goody 3, Kristin A. Gabor 4, Jacob R. Longfellow 1, Paul J. Millard 2,5, Carol H. Kim 1,2
1Department of Molecular and Biomedical Sciences, University of Maine, 2Graduate School of Biomedical Sciences and Engineering, University of Maine, 3School of Biology and Ecology, University of Maine, 4Division of Intramural Research, Immunity, Inflammation and Disease Laboratory, National Institute of Environmental Health Sciences, NIH, 5Department of Chemical and Biological Engineering, University of Maine

Systemic and localized zebrafish infection models for human influenza A virus are demonstrated. Using a systemic infection model, zebrafish can be used to screen antiviral drugs. Using a localized infection model, zebrafish can be used to characterize host immune cell responses.

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Environment

Impedance Pneumography for Minimally Invasive Measurement of Heart Rate in Late Stage Invertebrates
Amalia M. Harrington 1,2, Holland Haverkamp 1,3, Heather J. Hamlin 1,2
1School of Marine Sciences, University of Maine, 2Aquaculture Research Institute, University of Maine, 3Ecology and Environmental Sciences Program, University of Maine

Measuring heart rate during a thermal challenge provides insight into physiological responses of organisms as a consequence of acute environmental change. Using the American lobster (Homarus americanus) as a model organism, this protocol describes the use of impedance pneumography as a relatively noninvasive and nonlethal approach to measure heart rate in late stage invertebrates.

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Developmental Biology

A Drosophila Model to Study Wound-induced Polyploidization
Erin C. Bailey *1, Ari S. Dehn *1, Kayla J. Gjelsvik 2, Rose Besen-McNally 1, Vicki P. Losick 1
1Biology Department, Boston College, 2Graduate School of Biomedical Sciences and Engineering and Kathryn W. Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory, University of Maine

Wound-induced polyploidization is a conserved tissue repair strategy where cells grow in size instead of dividing to compensate for cell loss. Here is a detailed protocol on how to use the fruit fly as a model to measure ploidy and its genetic regulation in epithelial wound repair.

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Neuroscience

Lineage Tracing of Inducible Fluorescently-Labeled Stem Cells in the Adult Mouse Brain
Gabriel S. Jensen 1,2, Jake W. Willows 2, David T. Breault 3, Kristy L. Townsend 1,2
1Graduate School of Biomedical Science and Engineering, University of Maine, 2Department of Neurosurgery, Ohio State University Wexner Medical Center, 3Boston Children’s Hospital, Harvard Medical School

The ability to permanently mark stem cells and their progeny with a fluorophore using an inducible transgenic lineage tracing mouse line allows for spatial and temporal analysis of activation, proliferation, migration, and/or differentiation in vivo. Lineage tracing can reveal novel information about lineage commitment, response to intervention(s), and multipotency.

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