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Nat Methods. 2014 Mar;11(3):313-8. doi: 10.1038/nmeth.2835. Epub 2014 Feb 2.

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

Author information

1
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. [3].
2
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2].
3
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Program in Biophysics, Harvard University, Cambridge, Massachusetts, USA.
4
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA. [3] Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA.
5
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.

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.

PMID:
24487583
PMCID:
PMC4153392
DOI:
10.1038/nmeth.2835
[Indexed for MEDLINE]
Free PMC Article
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