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Membrane-enclosed structures called vesicles transport proteins and lipids across the cell. The vesicles derive their cargo from the plasma membrane, Golgi, ER, or endosome. Coated vesicles are spherical, protein-coated carriers with a 50–100 nm diameter that mediate bidirectional transport between the ER and the Golgi. The distribution of proteins between the ER and Golgi complex is dynamic and is maintained by different coated vesicles. Their formation is driven by the assembly of...
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Updated: Nov 23, 2025

Lipid Bilayer Vesicle Generation Using Microfluidic Jetting
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Making Droplet-Embedded Vesicles to Model Cellular Lipid Droplets.

Aymeric Chorlay1, Alexandre Santinho1, Abdou Rachid Thiam1

  • 1Laboratoire de Physique de l'École Normale Supérieure, ENS, Université PSL, CNRS, Sorbonne Université, Université de Paris, 75005 Paris, France.

STAR Protocols
|December 30, 2020
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Summary

We developed a protocol for droplet-embedded vesicles (DEVs), mimicking cellular lipid droplets (LDs) interacting with the endoplasmic reticulum (ER). This tunable model aids research into LD biogenesis and protein interactions at cellular interfaces.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Biophysics

Background:

  • Cellular lipid droplets (LDs) are crucial organelles involved in lipid metabolism and signaling.
  • The interaction between LDs and the endoplasmic reticulum (ER) membrane is vital for LD biogenesis and function.
  • Studying these interactions requires model systems that recapitulate cellular complexity with controllable parameters.

Purpose of the Study:

  • To present a reproducible protocol for creating droplet-embedded vesicles (DEVs).
  • To establish a model system that mimics the physical contact between cellular lipid droplets and the endoplasmic reticulum bilayer.
  • To enable the investigation of lipid droplet biogenesis and protein interactions at the ER-LD interface.

Main Methods:

  • Preparation of droplet-embedded vesicles (DEVs) using a phospholipid bilayer encapsulating an oil droplet.
  • Characterization of DEVs to confirm structural integrity and mimicry of cellular LD-ER interactions.
  • Utilizing DEVs as a platform for studying biophysical properties and molecular interactions.

Main Results:

  • Successfully developed a reproducible protocol for generating DEVs.
  • Demonstrated that DEVs effectively model the physical association of lipid droplets with endoplasmic reticulum membranes.
  • Established that the lipid composition and biophysical properties of both the droplet and bilayer are precisely controllable and tunable.

Conclusions:

  • Droplet-embedded vesicles (DEVs) provide a versatile and controllable model system for studying lipid droplet biology.
  • This model system facilitates research into the mechanisms of lipid droplet biogenesis and the factors governing protein binding at the ER-LD interface.
  • The protocol offers a valuable tool for advancing our understanding of lipid droplet-membrane interactions in a simplified, yet relevant, context.