Single-biomolecule observation with micro one-way valves for rapid buffer exchange

This paper describes a method for the rapid exchange of buffer solution during single-molecule observation. We use a simple, transparent, all-plastic one-way valve integrated on a coverslip on a flow cell. The valve is formed using a membrane made of parylene covering a microhole. It opens when a buffer solution is introduced from the microhole (the flow pushes the cover membrane) and closes when suctioning the buffer solution (the membrane is pulled back and seals the microhole to prevent the diffusion of the solution). To check valve performance, we observed the response of a rotary biomotor, $F1$-ATPase, for several buffer solutions in the fabricated chamber.