How to make and trap a pseudo-vesicle with a micropipette

TL;DR

This paper introduces a straightforward technique to create and manipulate giant lipid pseudo-vesicles within agarose gel using a micropipette, verified by fluorescence imaging and protein insertion.

Contribution

The method enables simple production and deformation of lipid pseudo-vesicles in gel, facilitating studies of membrane mechanics and protein interactions.

Findings

Successful formation of lipid pseudo-vesicles with an oily cap

Verification of lipid bilayer integrity via protein insertion

Mechanical deformation of vesicles demonstrated

Abstract

We present a simple method to produce giant lipid pseudo-vesicles (vesicles with an oily cap on the top), trapped in an agarose gel. The method can be implemented using only a regular micropipette and relies on the formation of a water/oil/water double droplet in liquid agarose. We characterize the produced vesicle with fluorescence imaging and establish the presence and integrity of the lipid bilayer by the successful insertion of {\alpha}-Hemolysin transmembrane proteins. Finally, we show that the vesicle can be easily mechanically deformed, non-intrusively, by indenting the surface of the gel.