Efficient Lipid Bilayer Formation by Dipping Lipid-Loaded Microperforated Sheet in Aqueous Solution

Koki Kamiya,Toshihisa Osaki,Shoji Takeuchi,Nobuo Misawa,Satoshi Fujii

doi:10.3390/mi12010053

Koki Kamiya, Toshihisa Osaki + Show 3 more

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https://doi.org/10.3390/mi12010053

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Abstract

This paper describes a method for a bilayer lipid membrane (BLM) formation using a perforated sheet along with an open chamber. Microscopic observation of the formed membrane showed a typical droplet interface bilayer. We proved that the formed membrane was a BLM based on electrical measurements of the membrane protein α-hemolysin, which produces nanopores in BLMs. Unlike the conventional approach for BLM formation based on the droplet contact method, this method provides aqueous surfaces with no organic solvent coating layer. Hence, this method is suitable for producing BLMs that facilitate the direct addition of chemicals into the aqueous phase.

Highlights

Bilayer lipid membranes (BLMs) are widely used as artificial cell membranes for research on plasma membranes, membrane proteins, and various membrane-associated biomolecules [1,2,3,4]
The system requires an accurate regulation of the solution injection and/or ejection to prevent damage to the BLMs owing to a pressure imbalance
We evaluated the stability of BLMs in our system basedononthe thetime timebetween betweenthe thefirst firstinsertion insertionofofαHL

Summary

Introduction

Bilayer lipid membranes (BLMs) are widely used as artificial cell membranes for research on plasma membranes, membrane proteins, and various membrane-associated biomolecules [1,2,3,4]. The “painting method” [10] and “monolayer-folding method” [11] have been frequently used and regarded as simple methods for obtaining BLMs. In recent years, the “droplet contact method” [12,13,14] has received significant attention for BLM production as an easier technique that generates a droplet interface bilayer (DIB) in organic solvents containing lipid molecules. This configuration makes it difficult to inject some chemicals, such as drug candidates; in addition, the aqueous phase from the outside impedes the fundamental studies of membrane proteins. The system requires an accurate regulation of the solution injection and/or ejection to prevent damage to the BLMs owing to a pressure imbalance

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Results

Conclusion