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7 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells
Martina Kučerová-Levisohn 1, Jordana Lovett 1, Armin Lahiji 1, Roxanne Holmes 2, Juan Carlos Zúñiga-Pflücker 2, Benjamin D. Ortiz 1
1Department of Biological Sciences, Hunter College and Graduate Center, City University of New York, 2Sunnybrook Research Institute, Department of Immunology, University of Toronto

Mouse embryonic stem cells can be differentiated to T cells in vitro using the OP9-DL1 co-culture system. Success in this procedure requires careful attention to reagent/cell maintenance, and key technique sensitive steps. Here we discuss these critical parameters and provide a detailed protocol to encourage adoption of this technology.

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Genetics

Manipulation of Ploidy in Caenorhabditis elegans
Erlyana K. Clarke 1, Katherine A. Rivera Gomez 1,2, Zaki Mustachi 1, Mikaela C. Murph 1,3, Mara Schvarzstein 1,2,3
1Brooklyn College, Biology Department, City University of New York, 2The Graduate Center, Biology Department, City University of New York, 3Advanced Science Research Center, City University of New York

This method allows for the generation of tetraploid and triploid Caenorhabditis nematodes from any diploid strain. Polyploid strains generated by this method have been used to study chromosome interactions in meiotic prophase, and this method is useful for examining important basic questions in cell, developmental, evolutionary, and cancer biology.

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JoVE Core

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers
Irina E. Catrina 1, Livia V. Bayer 1,2, Omar S. Omar 1,2, Diana P. Bratu 1,2
1Biological Sciences Department Hunter College, City University of New York, 2Program in Molecular, Cellular, and Developmental Biology, Graduate Center, City University of New York

Here, we present a protocol for the visualization, detection, analysis and tracking of endogenous mRNA trafficking in live Drosophila melanogaster egg chamber using molecular beacons, spinning disc confocal microscopy, and open-source analysis software.

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Behavior

Lexical Decision Task for Studying Written Word Recognition in Adults with and without Dementia or Mild Cognitive Impairment
Alexandre Nikolaev 1, Eve Higby 2,3, JungMoon Hyun 4, Sameer Ashaie 5,6
1Helsinki Collegium for Advanced Studies, University of Helsinki, 2University of California, Riverside, 3California State University, East Bay, 4Hunter College, City University of New York, 5Feinberg School of Medicine, Northwestern University, 6Shirley Ryan AbilityLab

This article describes how to implement a simple lexical decision experiment to assess written word recognition in neurologically healthy participants and in individuals with dementia and cognitive decline. We also provide a detailed description of reaction time analysis using principal components analysis (PCA) and mixed-effects modeling.

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Genetics

2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
Ning Zhang 1,2, Shundi Shi 2, Barney Yoo 3, Xiaohong Yuan 1, Wenjia Li 4, Shenglong Zhang 1
1Department of Biological and Chemical Sciences, New York Institute of Technology, 2Department of Chemical Engineering, Columbia University, 3Department of Chemistry, Hunter College, City University of New York, 4Department of Computer Science, New York Institute of Technology

Here, we describe a detailed protocol for an LC-MS-based sequencing method that can be used as a direct method to sequence short RNA (<35 nt per run) without a cDNA intermediate, and as a general method to sequence different nucleotide modifications in a single study at single-base precision.

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Environment

Simulating Impacts of Ice Storms on Forest Ecosystems
John L. Campbell 1, Lindsey E. Rustad 1, Charles T. Driscoll 2, Ian Halm 3, Timothy J. Fahey 4, Habibollah Fakhraei 5, Peter M. Groffman 6,7, Gary J. Hawley 8, Wendy Leuenberger 9, Paul G. Schaberg 10
1Northern Research Station, U.S. Forest Service, Durham, NH, 2Department of Civil and Environmental Engineering, Syracuse University, 3Northern Research Station, U.S. Forest Service, North Woodstock, NH, 4Department of Natural Resources, Cornell University, 5Department of Civil and Environmental Engineering, Southern Illinois University, 6Advanced Science Research Center at the Graduate Center, City University of New York, 7Cary Institute of Ecosystem Studies, 8Rubenstein School of Environment and Natural Resources, University of Vermont, 9Department of Forestry and Natural Resources, University of Kentucky, 10Northern Research Station, U.S. Forest Service, Burlington, VT

Ice storms are important weather events that are challenging to study because of difficulties in predicting their occurrence. Here, we describe a novel method for simulating ice storms that involves spraying water over a forest canopy during sub-freezing conditions.

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Experimental Entomology in the Age of Video
Daniel A. Friedman 1, Judith R. Wexler 2, Sebastian Alvarado 3
1Department of Entomology & Nematology, University of California, 2Department of Ecology, Evolution, and Behavior, The Hebrew University in Jerusalem, 3Department of Biology, Queens College, City University of New York

Experimental Entomology in the Age of Video

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