Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT.
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.
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. [3].
Ralf
R
Jungmann
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. [3].
Maier S
MS
Avendaño
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2].
Johannes B
JB
Woehrstein
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Program in Biophysics, Harvard University, Cambridge, Massachusetts, USA.
Mingjie
M
Dai
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.
William M
WM
Shih
1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. [2] Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
Peng
P
Yin
D003116:Color; D003584:Cytological Techniques / Q000379:methods*; D004247:DNA / Q000737:chemistry; D021621:Imaging, Three-Dimensional; D008856:Microscopy, Fluorescence / Q000639:trends*
D004247:DNA