Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT
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Open Access
- 2 February 2014
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Methods
- Vol. 11 (3), 313-318
- https://doi.org/10.1038/nmeth.2835
Abstract
Variations on point accumulation for imaging in nanoscale topography (PAINT) for super-resolution imaging extend the technology to allow simple imaging of cellular proteins as well as synthetic DNA nanostructures and provide a high level of multiplexing. 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.Keywords
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