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

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 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.

SUBMITTER: Jungmann R

PROVIDER: S-EPMC4153392 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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

Jungmann Ralf R Avendaño Maier S MS Woehrstein Johannes B JB Dai Mingjie M Shih William M WM Yin Peng P

Nature methods 20140202 3

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. ...[more]

PMID: 24487583

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