DNA-Paint and Exchange-Paint for Multiplexed 3D Super-Resolution Microscopy

Abstract

While it's clear that technological and scientific advances often go hand-in-hand, it's generally difficult to identify which combinations will produce the most lasting impact. Here, we present an unlikely union of DNA engineering and optical microscopy that enables 3D super-resolution imaging with extreme color multiplexing [1]. In essence, DNA-PAINT utilizes thermally-driven binding and unbinding of engineered DNA sequences to induce fluorophore blinking, which ultimately yields images with sub-10-nm spatial resolution. Moreover, because binding sites and flurophores are connected on opposite strands of hybridized DNA, careful engineering of the base pair sequence allows for orthogonal selectivity between signal and target. Thus, EXCHANGE-PAINT goes beyond the single color detection of DNA-PAINT and enables an extreme form of color multiplexing. By combining this unique approach to target labeling with high-speed confocal imaging, we are now in position to move toward full 3D super-resolution imaging. In this work, we present recent progress toward this ultimate goal and demonstrate how such techniques can provide unique insights on molecular scale biophysical systems. R.J., M.D., M.S.A., J.B.W. and P.Y. have filed a provisional US patent application regarding the current work. [1] R. Jungmann, M.S. Avendaño, J.B. Woehrstein, M. Dai. W.M. Shih, P. Yin. Multiplexed Cellular 3D Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT. Nature Methods 11, 313-318 (2014).