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JoVE Journal Neuroscience
SNARE-medieret Fusion af Single proteoliposomer med bundet understøttede dobbeltlag i en Mikrofluid Flow Cell Overvåget af Polariseret TIRF Microscopy
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SNARE-medieret Fusion af Single proteoliposomer med bundet understøttede dobbeltlag i en Mikrofluid Flow Cell Overvåget af Polariseret 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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Her præsenterer vi en protokol til at opdage enkelt, SNARE-medieret fusion begivenheder mellem liposomer og understøttede dobbeltlag i mikrofluide kanaler ved hjælp polariseret TIRFM, med enkelt molekyle følsomhed og ~ 15 ms tidsopløsning. Lipid- og opløseligt fragt frigivelse kan detekteres samtidigt. Liposomstørrelse, lipid diffusivitet, og fusion pore egenskaber måles.

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Neuroscience Membrane fusion SNARE proteiner lipid dobbeltlag proteoliposomer støttet dobbeltlag enkelt vesikel fusion fusion pore exocytose single-molekyle fluorescens mikrofluidik TIRFM biofysik
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