Bottom-Up In Vitro Methods to Assay the Ultrastructural Organization, Membrane Reshaping, and Curvature Sensitivity Behavior of Septins

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2.3K Views

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09:09 min

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August 17th, 2022

DOI :

10.3791/63889-v

August 17th, 2022

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Brieuc Chauvin*¹, Koyomi Nakazawa*¹, Alexandre Beber¹^,⁷, Aurélie Di Cicco¹, Bassam Hajj¹, François Iv², Manos Mavrakis², Gijsje H. Koenderink³, João T. Cabral⁴, Michaël Trichet⁵, Stéphanie Mangenot*⁶, Aurélie Bertin*¹

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

Podsumowanie

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Bottom up Methods

Cytoskeletal Filaments

Curvature Sensitivity

Septins

In Vitro Assays

Scanning Electron Microscopy

Norland Optical Adhesive

PDMS Template

Small Unilamellar Vesicles

Supported Lipid Bilayer

Glutaraldehyde Fixation

UV Photopolymerization

Air Plasma Cleaning

Lipid Film Resuspension

Rozdziały w tym wideo

0:05

Introduction

0:49

Designing Wavy NOA (Norland Optical Adhesive) Replica from Wavy PDMS (Polydimethylsiloxane) Patterns

1:49

Generation of a Supported Lipid Bilayer and Protein Incubation

3:22

Sample Fixation, Sample Dehydration, Critical Point Drying

4:43

Sample Mounting and Coating

5:54

Acquire the Images Using a Field-Emission SEM (FE-SEM) Microscope

6:56

Results: Analysis of the Self-Assembly of Septins at the Lipid Membrane

8:27

Conclusion

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