Automated Lipid Bilayer Membrane Formation Using a Polydimethylsiloxane Thin Film

J Vis Exp. 2016 Jul 10:(113):54258. doi: 10.3791/54258.

Abstract

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for biosensor applications. However, conventional BLM formation techniques have several drawbacks and they often require specific expertise and laborious processes. In particular, conventional BLMs suffer from low formation success rates and inconsistent membrane formation time. Here, we demonstrate a storable and transportable BLM formation system with controlled thinning-out time and enhanced BLM formation rate by replacing conventionally used films (polytetrafluoroethylene, polyoxymethylene, polystyrene) to polydimethylsiloxane (PDMS). In this experiment, a porous-structured polymer such as PDMS thin film is used. In addition, as opposed to conventionally used solvents with low viscosity, the use of squalene permitted a controlled thinning-out time via slow solvent absorption by PDMS, prolonging membrane lifetime. In addition, by using a mixture of squalene and hexadecane, the freezing point of the lipid solution was increased (~16 °C), in addition, membrane precursors were produced that can be indefinitely stored and readily transported. These membrane precursors have reduced BLM formation time of < 1 hr and achieved a BLM formation rate of ~80%. Moreover, ion channel experiments with gramicidin A demonstrated the feasibility of the membrane system.

Publication types

  • Video-Audio Media

MeSH terms

  • Biosensing Techniques
  • Dimethylpolysiloxanes / chemistry*
  • Gramicidin / chemistry
  • Ion Channels / chemistry
  • Lipid Bilayers / chemical synthesis*
  • Lipid Bilayers / chemistry*

Substances

  • Dimethylpolysiloxanes
  • Ion Channels
  • Lipid Bilayers
  • Gramicidin