Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT.

Autor:	Schueder F; Department of Physics and Center for Nanoscience, Ludwig Maximilian University, 80539, Munich, Germany.; Max Planck Institute of Biochemistry, 82152, Martinsried, Germany., Lara-Gutiérrez J; Wyss Institute for Biologically Inspired Engineering, Boston, MA, 02115, USA.; Department of Systems Biology, Harvard University, Boston, MA, 02115, USA., Beliveau BJ; Wyss Institute for Biologically Inspired Engineering, Boston, MA, 02115, USA.; Department of Systems Biology, Harvard University, Boston, MA, 02115, USA., Saka SK; Wyss Institute for Biologically Inspired Engineering, Boston, MA, 02115, USA.; Department of Systems Biology, Harvard University, Boston, MA, 02115, USA., Sasaki HM; Wyss Institute for Biologically Inspired Engineering, Boston, MA, 02115, USA.; Department of Systems Biology, Harvard University, Boston, MA, 02115, USA., Woehrstein JB; Department of Physics and Center for Nanoscience, Ludwig Maximilian University, 80539, Munich, Germany.; Max Planck Institute of Biochemistry, 82152, Martinsried, Germany., Strauss MT; Department of Physics and Center for Nanoscience, Ludwig Maximilian University, 80539, Munich, Germany.; Max Planck Institute of Biochemistry, 82152, Martinsried, Germany., Grabmayr H; Department of Physics and Center for Nanoscience, Ludwig Maximilian University, 80539, Munich, Germany.; Max Planck Institute of Biochemistry, 82152, Martinsried, Germany., Yin P; Wyss Institute for Biologically Inspired Engineering, Boston, MA, 02115, USA. py@hms.harvard.edu.; Department of Systems Biology, Harvard University, Boston, MA, 02115, USA. py@hms.harvard.edu., Jungmann R; Department of Physics and Center for Nanoscience, Ludwig Maximilian University, 80539, Munich, Germany. jungmann@biochem.mpg.de.; Max Planck Institute of Biochemistry, 82152, Martinsried, Germany. jungmann@biochem.mpg.de.
Jazyk:	angličtina
Zdroj:	Nature communications [Nat Commun] 2017 Dec 12; Vol. 8 (1), pp. 2090. Date of Electronic Publication: 2017 Dec 12.
DOI:	10.1038/s41467-017-02028-8
Abstrakt:	Single-molecule localization microscopy (SMLM) can visualize biological targets on the nanoscale, but complex hardware is required to perform SMLM in thick samples. Here, we combine 3D DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) with spinning disk confocal (SDC) hardware to overcome this limitation. We assay our achievable resolution with two- and three-dimensional DNA origami structures and demonstrate the general applicability by imaging a large variety of cellular targets including proteins, DNA and RNA deep in cells. We achieve multiplexed 3D super-resolution imaging at sample depths up to ~10 µm with up to 20 nm planar and 80 nm axial resolution, now enabling DNA-based super-resolution microscopy in whole cells using standard instrumentation.
Databáze:	MEDLINE
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