Recently biological imaging techniques have been rapidly advanced. In particular, real-time visualization at the single-molecule level has been making a big break-through in life sciences. Here we introduce a novel single-molecule imaging technique DNA curtain that is a hybrid system concatenating nano-fabrication, microfluidics, lipid fluidity, and fluorescence microscopy. The DNA curtain system provides a universal platform to study diverse protein-DNA interaction in a direct manner. Using the DNA curtain, we investigate how DNA damage repair proteins search for DNA lesions.