CPM Seminar

Approaching the limit: Multiplexed Super-Resolution Microscopy with DNA-PAINT and Exchange-PAINT

Ralf Jungmann

Wyss Institute
Harvard University

Super-resolution fluorescence microscopy is a powerful tool for biological research, but obtaining multiplexed images for a large number of distinct target species remains challenging. Here we use the transient binding of short fluorescently labeled oligonucleotides (DNA-PAINT, a variation of point accumulation for imaging in nanoscale topography) for simple and easy-to-implement multiplexed superresolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures.

We report a multiplexing approach (Exchange-PAINT) that allows sequential imaging of multiple targets using only a single dye and a single laser source. We experimentally demonstrate ten-color super-resolution imaging in vitro on synthetic DNA structures as well as four-color two-dimensional imaging and three-color 3D imaging of proteins in fixed cells.

References:
1. R. Jungmann, M.S. Avenda�o, J.B. Woehrstein, M. Dai, W.M. Shih, P. Yin, Nature Methods (2014).
2. R. Iinuma, Y. Ke, R. Jungmann, T. Schlichthaerle, J.B. Woehrstein, P. Yin. Science (2014).
3. N.D. Derr, B.S. Goodman, R. Jungmann, A.E. Leschziner, W.M. Shih, S.L. Reck-Peterson. Science (2012).
4. C. Lin, R. Jungmann, A.M. Leifer, C. Li, D. Levner, G.M. Church, W.M. Shih, and P. Yin. Nature Chemistry (2012).
5. R. Jungmann, C. Steinhauer, M. Scheible, A. Kuzyk, P. Tinnefeld, and F.C. Simmel. Nano Letters (2010).