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Trinity College Dublin

8 ARTICLES PUBLISHED IN JoVE

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Biology

Crystallizing Membrane Proteins for Structure Determination using Lipidic Mesophases
Martin Caffrey 1, Christopher Porter 1
1Membrane Structural and Functional Biology Group, Schools of Biochemistry and Immunology and Medicine, Trinity College Dublin

Herein is described the procedure implemented in the Caffrey Membrane Structural and Functional Biology Group to set up manually crystallization trials of membrane proteins in lipidic mesophases.

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Biology

Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases
Dianfan Li 1, Coilín Boland 1, Kilian Walsh 1, Martin Caffrey 1
1Membrane Structural and Functional Biology Group, Schools of Medicine and Biochemistry & Immunology, Trinity College Dublin

Herein is described a robotic approach to high-throughput crystallization of membrane proteins in lipidic mesophases for use in structure determination using macromolecular X-ray crystallography. Three robots capable of handling the viscous and sticky protein-laden mesophase integral to the method are introduced.

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Biology

Harvesting and Cryo-cooling Crystals of Membrane Proteins Grown in Lipidic Mesophases for Structure Determination by Macromolecular Crystallography
Dianfan Li 1, Coilín Boland 1, David Aragao 1, Kilian Walsh 1, Martin Caffrey 1
1Membrane Structural and Functional Biology Group, Schools of Medicine and Biochemistry & Immunology, Trinity College Dublin

Herein is described procedures implemented in the Caffrey Membrane Structural and Functional Biology Group to harvest and cryo-cool membrane protein crystals grown in lipidic cubic and sponge phases for use in structure determination using macromolecular X-ray crystallography.

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Bioengineering

Harmonic Nanoparticles for Regenerative Research
Flavio Ronzoni 1, Thibaud Magouroux 2, Remi Vernet 1, Jérôme Extermann 3, Darragh Crotty 4, Adriele Prina-Mello 5, Daniel Ciepielewski 6, Yuri Volkov 4, Luigi Bonacina 2, Jean-Pierre Wolf 2, Marisa Jaconi 1
1Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, 2Physics Department, GAP-Biophotonics, University of Geneva, 3Laboratoire d'Optique Biomédicale (LOB), Faculté des Sciences et Techniques de l'Ingénieur, École Polytechnique Fédérale de Lausanne, 4Department of Clinical Medicine, School of Medicine, Trinity College Dublin, 5School of Medicine and CRANN, Trinity College Dublin, 6Nikon AG Instruments

Protocol details are provided for in vitro labeling human embryonic stem cells with second harmonic generating nanoparticles. Methodologies for hESC investigation by multi-photon microscopy and their differentiation into cardiac clusters are also presented.

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Medicine

Sublingual Immunotherapy as an Alternative to Induce Protection Against Acute Respiratory Infections
Natalia Muñoz-Wolf 1,2, Analía Rial 1, José M. Saavedra 1, José A. Chabalgoity 1
1Departamento de Desarrollo Biotecnológico, Universidad de la República, 2Present Affiliation: Trinity Biomedical Science Institute, Trinity College Dublin

The present work illustrates the convenience of using sublingual immunotherapy to boost the innate immune response in the lungs and confer protection against acute pneumococcal pneumonia in mouse.

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Engineering

Preparation of Liquid-exfoliated Transition Metal Dichalcogenide Nanosheets with Controlled Size and Thickness: A State of the Art Protocol
Claudia Backes 1, Damien Hanlon 2, Beata M. Szydlowska 2, Andrew Harvey 2, Ronan J. Smith 2, Thomas M. Higgins 1, Jonathan N. Coleman 2
1Chair of Applied Physical Chemistry, Ruprecht-Karls University Heidelberg, 2School of Physics and CRANN, Trinity College Dublin

A protocol for the liquid exfoliation of layered materials to nanosheets, their size selection and size measurement by microscopic and spectroscopic techniques is presented.

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Developmental Biology

Serum Free Production of Three-dimensional Human Hepatospheres from Pluripotent Stem Cells
Balta Lucendo-Villarin 1, Hassan Rashidi 1,2, Sharmin Alhaque 1,3, Lena Fischer 1,4, Jose Meseguer-Ripolles 1, Yu Wang 1, Cliona O'Farrelly 4, Michael Themis 3, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh, 2UCL Great Ormond Street Institute of Child Health, University College London, 3Division of Biosciences, Department of Life Sciences, College of Health and Life Sciences, Brunel University London, 4School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin

This protocol describes an approach to produce hepatospheres from human pluripotent stem cells using a defined culture system and cell self-assembly. This protocol is reproducible in a number of cell lines, cost effective and allows the production of stable human hepatospheres for biomedical application.

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Environment

Sampling, Identification and Characterization of Microplastics Release from Polypropylene Baby Feeding Bottle during Daily Use
Dunzhu Li *1,2, Luming Yang *1,2, Rachel Kavanagh 1, Liwen Xiao 2,3, Yunhong Shi 1,2, Daniel K. Kehoe 1, Emmet D. Sheerin 1,4, Yurii K. Gun’ko 4,5, John J. Boland 1,4, Jing Jing Wang 1
1AMBER Research Centre and Centre for Research on Adaptive Nanostructures and Nanodevices (CRANN), Trinity College Dublin, 2Department of Civil, Structural and Environmental Engineering, Trinity College Dublin, 3TrinityHaus, Trinity College Dublin, 4School of Chemistry, Trinity College Dublin, 5BEACON, Bioeconomy SFI Research Centre, University College Dublin

This study detailed a reliable and cost-effective protocol for microplastics collection and detection from the daily use of plastic products.

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