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

Joerg Nikolaus; Erdem Karatekin

doi:10.3791/54349

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

JoVE Journal
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

10,898 Views

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10:58 min

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August 24, 2016

DOI:

10.3791/54349-v

DOI:

10.3791/54349-v

•

10:58 min

August 24, 2016

•

10881 Views

Joerg Nikolaus^1,2, Erdem Karatekin^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)

Summary

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.