Easy and stable lipid bilayer formation: A droplets-contacting-method in parylene mircopores for multiple ion channel recordings

摘要

This paper describes simultaneou us 16 channels ion-channel recordings through membrane proteins reconstituted in bilayer lipid membrannes (BLMs) as shown in Figure 1. Although multiple ion-channel recording is necessary for high-throughput screening (HTS) of drug [1], there are no reports of such systems as far as the authors are aware. We have developed several strategies to construct BLMs arrray for multiple ion-channel recording. The “droplets contacting method” which forms BLMs at the interface of two contacting droplets of lipid monolayers is one of the simplest and most efficient methods. Since it does not require skilled techniques, it is highly reproducible and can be applied to an automated system. [2]. In this study we particularly used a double well chip (DWC) (Figure 1a [3]) with parylene micro-pores in thedroplets contacting method. The parylene micro-pores confined the BLMs forming areas and subsequently, augmented the mechanical stability [4]. Using the droplets contacting method and parylene micro-pores, we succeeded in the multiple recording in 14 channels, which is the most multiple recording as far as we know. This method can be applied to fully automated membrane protein reconstitution for HTS system using a spotting robot [5].