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Neuroscience

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JoVE Journal Neuroscience
SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy
 

SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy

Article DOI: 10.3791/54349-v • 10:58 min • August 24th, 2016
August 24th, 2016 •
1,2, 1,2,3,4
1Department of Cellular and Molecular Physiology, Yale University School of Medicine, 2Nanobiology Institute, Yale University, 3Department of Molecular Biophysics and Biochemistry, Yale University, 4Laboratoire de Neurophotonique, Université Paris Descartes, Faculté des Sciences Fondamentales et Biomédicales, Centre National de la Recherche Scientifique (CNRS)

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Here, we present a protocol to detect single, SNARE-mediated fusion events between liposomes and supported bilayers in microfluidic channels using polarized TIRFM, with single molecule sensitivity and ~15 msec time resolution. Lipid and soluble cargo release can be detected simultaneously. Liposome size, lipid diffusivity, and fusion pore properties are measured.

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SNARE-mediated Fusion Proteoliposomes Tethered Supported Bilayers Microfluidic Flow Cell Polarized TIRF Microscopy Membrane Fusion Events Neuronal SNARE Proteins Exocytotic SNARE Proteins Neurotransmitter Release Hormone Release Fusion Pore Cargo Release Kinetics Vesicle Docking Biochemically Defined Environments Exocytosis Envelope Viruses Target Cell Membranes Infection Microfabrication Protein Purification Imaging
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