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1Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University,Sorbonne Université, 2Institut Fresnel, CNRS UMR7249,Aix Marseille Univ, Centrale Marseille, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft,Delft University of Technology, 4Department of Chemical Engineering,Imperial College London, 5Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS),Service de microscopie électronique (IBPS-SME), 6Laboratoire Matière et Systèmes Complexes (MSC),Université Paris Cité, 7Institute of Biotechnology,Czech Academy of Sciences, BIOCEV
Chapters
- 00:05Introduction
- 00:49Designing Wavy NOA (Norland Optical Adhesive) Replica from Wavy PDMS (Polydimethylsiloxane) Patterns
- 01:49Generation of a Supported Lipid Bilayer and Protein Incubation
- 03:22Sample Fixation, Sample Dehydration, Critical Point Drying
- 04:43Sample Mounting and Coating
- 05:54Acquire the Images Using a Field-Emission SEM (FE-SEM) Microscope
- 06:56Results: Analysis of the Self-Assembly of Septins at the Lipid Membrane
- 08:27Conclusion
Septins are cytoskeletal proteins. They interact with lipid membranes and can sense but also generate membrane curvature at the micron scale. We describe in this protocol bottom-up in vitro methodologies for analyzing membrane deformations, curvature-sensitive septin binding, and septin filament ultrastructure.
