2013YWL

黎彥汶 (Yan-Wen Li)

Construction of An Asymmetric Lipid Bilayer Formation System For Atomic Force Microscopy Measurement

可配合原子力顯微鏡量測之非對稱脂質雙層膜成膜平台的建構

Lipid bilayer membrane is very common in organism, and it is a very important component of a biological cell. Most of lipid bilayer membranes are asymmetric, such as the membrane of erythrocyte. The properties of artificial asymmetric bilayer are more like the original cell membrane. When protein or some cell components work on it, the behavior of the component will more close to biological organism in nature.

This lipid bilayer formation system is based upon connecting aqueous volumes immersed in a hydrophobic phase containing lipids. Briefly, a lipid monolayer self-assembles at the water-oil boundary of the aqueous components, and when two components are brought into contact, a bilayer spontaneously forms between them. This formation system and method have several advantages. Full-oil-phase environment prevent aqueous droplet evaporation in oil chamber. To increase the lifetime of bilayer, hydrogel-supporting design makes bilayer strong. In experiments, the symmetric bilayer has been formed to discuss the factors that might afford stability and lifetime of bilayer. In bilayer formation experiment with optical image, the optical image was viewed by using inverted fluorescence microscopy, and formation success rate is 83%. The formation and lifetime of bilayer recorded in video clip through CCD. The longest lifetime of bilayer is 15 minutes. The electrophysiological recording measures by using patch clamp. The electrophysiological signal was detected through all formation timeline, and the longest lifetime of bilayer is about 1 minute. Bilayer capacitance per area is 0.32 μF/cm2.

The lipid bilayer formation system will soon to be use to form asymmetric bilayer. Further more, membrane also will insert into the device to record ion channel signals, this can make sure the feasibility of the system. The final goal is that using atomic force microscopy to measure the dynamic situation when proteins are working, quantify the values of protein’s displacement.