Jungmann, RalfAvendaño, Maier SWoehrstein, Johannes BDai, MingjieShih, WilliamYin, Peng2023-02-132014Jungmann, Ralf, Maier S Avendaño, Johannes B Woehrstein, Mingjie Dai, William M Shih, and Peng Yin. 2014. “Multiplexed 3D Cellular Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT.” Nature Methods 11 (3) (February 2): 313–318. doi:10.1038/nmeth.2835.1548-70911548-7105https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37374349Super-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 super-resolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures. We also 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 (2D) imaging and three-color 3D imaging of proteins in fixed cells.en-USSuper-resolution microscopyFluorescence imagingCellular imagingJournal Article2023-02-1310.1038/nmeth.2835