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SNARE-medieret Fusion af Single proteoliposomer med bundet understøttede dobbeltlag i en Mikrofluid Flow Cell Overvåget af Polariseret TIRF Microscopy
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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)
Chapters
- 00:05Title
- 01:01Preparation of a PDMS Block to Form the Microfluidic Channel
- 03:17Formation of the Tethered Supported Bilayer
- 06:18Checking Bilayer Fluidity
- 07:06Introducing v-SUVs into the Microfluidic Channels
- 07:59Observing the Fusion of Single v-SUVs and the SBL
- 08:42Results: Docking and Fusion Events Described by Lipid and Content Release
- 10:26Conclusion
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.
