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Simon Fraser University

11 ARTICLES PUBLISHED IN JoVE

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Biology

Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons
David M. Kwinter 1, Michael A. Silverman 1
1Department of Biological Sciences, Simon Fraser University

Live cell imaging is of particular utility when studying the dynamics of organelle trafficking. Here we describe a protocol for live imaging of dense-core vesicles in cultured neurons using wide-field fluorescence microscopy. This protocol is flexible and can be adapted to image other organelles such as mitochondria, endosomes, and peroxisomes.

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Erratum: Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons.
David M. Kwinter 1, Michael A. Silverman 1
1Department of Biological Sciences, Simon Fraser University

A correction to author names has been made for article Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons.

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Biology

Glycopeptide Capture for Cell Surface Proteomics
M. C. Gilbert Lee 1, Bingyun Sun 1
1Department of Chemistry, Simon Fraser University

Cell surface proteins are biologically important and widely glycosylated. We introduce here a glycopeptide-capture approach to solubilize, enrich, and deglycosylate these proteins for facile LC-MS based proteomic analyses.

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Neuroscience

Detection of In Situ Protein-protein Complexes at the Drosophila Larval Neuromuscular Junction Using Proximity Ligation Assay
Simon Wang 1, SooHyun Yoo 1, Hae-yoon Kim 1, Mannan Wang 2, Clare Zheng 2, Wade Parkhouse 2, Charles Krieger 2, Nicholas Harden 1
1Department of Molecular Biology and Biochemistry, Simon Fraser University, 2Department of Biomedical Physiology and Kinesiology, Simon Fraser University

This protocol demonstrates how Proximity Ligation Assay can be used to detect in situ protein-protein interactions at the Drosophila larval neuromuscular junction. With this technique, Discs large and Hu-li tai shao are shown to form a complex at the postsynaptic region, an association previously identified through co-immunoprecipitation.

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Medicine

Busulfan as a Myelosuppressive Agent for Generating Stable High-level Bone Marrow Chimerism in Mice
Kyle Peake 1, John Manning 1, Coral-Ann Lewis 1,2, Christine Barr 1, Fabio Rossi 2, Charles Krieger 1,3
1Department of Biomedical Physiology and Kinesiology, Simon Fraser University, 2The Biomedical Research Centre, University of British Columbia, 3Division of Neurology, Department of Medicine, Neuromuscular Disease Unit, VHHSC

We describe a protocol whereby busulfan conditioning permits the bone marrow of a recipient mouse to be replaced with bone marrow cells from donor mice ubiquitously expressing green fluorescent protein, in the absence of irradiation. This technique is useful to study bone marrow cell accumulation in the central nervous system.

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Medicine

Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis
Winnie Enns 1, Anna von Rossum 1, Jonathan Choy 1
1Department of Molecular Biology and Biochemistry, Simon Fraser University

We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure.

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Chemistry

A 'Plug and Play' Method to Create Water-dispersible Nanoassemblies Containing an Amphiphilic Polymer, Organic Dyes and Upconverting Nanoparticles
Khaled M. Arafeh *1, Amir M. Asadirad *1, Jason Woodson Li 1, Danielle Wilson *1, Tuoqi Wu *1, Neil R. Branda 1
14D LABS and Department of Chemistry, Simon Fraser University

Organic dye molecules and oleic acid coated upconverting nanoparticles are not water-soluble. This protocol describes a ‘plug and play’ method that enables the transfer of organic dye molecules and upconverting particles from their initial hydrophobic solvent to water.

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JoVE Core

Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields
Seung C. Kim 1, Alex Mason 2, Wooseok Im 3
1College of Medicine, Yonsei University, 2Simon Fraser University, 3Biomedical Research Institute, Seoul National University Hospital

The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.

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Environment

Determination of the Settling Rate of Clay/Cyanobacterial Floccules
Tiffany Playter 1, Kurt Konhauser 1, George W. Owttrim 2, Denise S. Whitford 2, Tyler Warchola 1, Cheryl Hodgson 1,3, Aleksandra M. Mloszewska 4, Bruce Sutherland 1, J.-P. Zonneveld 1, S. George Pemberton 1, Murray K. Gingras 1
1Department of Earth and Atmospheric Sciences, University of Alberta, 2Department of Biological Sciences, University of Alberta, 3Department of Earth Sciences, Simon Fraser University, 4Earth Sciences Department, University of Toronto

The interaction and sedimentation of the clay and bacterial cells within the marine realm, observed in natural environments, can be best investigated in a controlled lab environment. Here, we describe a detailed protocol, which outlines a novel method for measuring the sedimentation rate of clay and cyanobacterial floccules.

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Biochemistry

Fluorescent Visualization of Mango-tagged RNA in Polyacrylamide Gels via a Poststaining Method
Iqra M. Yaseen *1, Quiana R. Ang *1, Peter J. Unrau 1
1Department of Molecular Biology and Biochemistry, Simon Fraser University

Here we present a sensitive, rapid, and discriminating post-gel staining method to image RNAs tagged with RNA Mango aptamers I, II, III, or IV, using either native or denaturing polyacrylamide gel electrophoresis (PAGE) gels. After running standard PAGE gels, Mango-tagged RNA can be easily stained with TO1-Biotin and then analyzed using commonly available fluorescence readers.

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Biology

CRISPR-Cas9-Mediated Precise Knock-In Edits in Zebrafish Hearts
Kyle E. Simpson 1, Shoaib Faizi 1, Ravichandra Venkateshappa 1, Mandy Yip 1, Raj Johal 1, Damon Poburko 1, Yen May Cheng 1, Diana Hunter 1, Eric Lin 1, Glen F. Tibbits 1, Thomas W. Claydon 1
1Department of Biomedical Physiology and Kinesiology, Simon Fraser University

This protocol describes an approach to facilitate precise knock-in edits in zebrafish embryos using CRISPR-Cas9 technology. A phenotyping pipeline is presented to demonstrate the applicability of these techniques to model a Long QT Syndrome-associated gene variant.

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