Banneyake, Bm U. (2008-12). Development of a Microfluidic Device for Synthesis of Lipid Bi-Layer In-Situ. Master's Thesis.
Lipid bi-layers are ubiquitous components of biological cells and are found in a variety of cell components. In biological membranes, lipid bi-layer membranes carry membrane proteins, which control transport of material and communication of signals in and out the cell. There are several disadvantages involved with patch clamping method as a way of studying biological membranes and protein interactions. Hence, artificial synthesis of bi-layer has been of great interest in basic biophysical studies, drug discoveries in pharmaceutical studies and study of protein nanopores for precise engineering applications. However, conventional lipid bi-layer synthesis techniques require skilled operators, have low repeatability (reliability), have portability restrictions and result in unstable bi-layers having a short lifetime. In this investigation a novel microfluidic device and a method for artificial synthesis of lipid bi-layer in-situ are explored. In the proposed method, lipid trapped at an aperture on a Teflon sheet, is thinned to form a lipid bi-layer by a continuous flow of buffer solution on both sides of the aperture in the microfluidic device. The microfluidic device is expected to have advantages from its compact design. Further, the new approach is expected to be repetitive and good for automation removing the requirement of a skilled operator. The microfluidic device was fabricated using two glass substrates. Two channels of ? ? shape were etched and through holes were fabricated at all four terminal ends of the microchannels on each glass substrate. A thin Teflon sheet carrying a 100?m diameter hole was sandwiched between the two glass wafers forming two sets of microchannels on both sides of the aperture. An analytical microfluid model of the microchannels was developed to investigate the nature of the flow and to select microchannel parameters. Experiments using the proposed device were performed to verify the feasibility of the novel approach for lipid bi-layer synthesis. Experimental results suggest formation of a lipid bi-layer at an aperture on the Teflon sheet but further investigation might be necessary for verification. Life time of the bi-layer is short mainly due to low quality of the used aperture.