Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT

Ralf Jungmann, Maier S. Avendano, Johannes B Woehrstein, Mingjie Dai, William M Shih, Peng Yin

Nature Methods 11(3):313-318 (2014).

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Abstract:   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 super-resolution imaging that achieves sub-10-nm spatial resolution in vitro on synthetic DNA structures. We also report a multiplexing apporach (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.

   

  • Summary: This ultrasharp Exchange-PAINT image simultaneously spots microtubules (green), mitochondria (purple), Golgi apparatus (red), and peroxisomes (yellow) from a single human cell.

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