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

Author

Listed:
  • Florian Schueder

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Juanita Lara-Gutiérrez

    (Wyss Institute for Biologically Inspired Engineering
    Harvard University)

  • Brian J. Beliveau

    (Wyss Institute for Biologically Inspired Engineering
    Harvard University)

  • Sinem K. Saka

    (Wyss Institute for Biologically Inspired Engineering
    Harvard University)

  • Hiroshi M. Sasaki

    (Wyss Institute for Biologically Inspired Engineering
    Harvard University)

  • Johannes B. Woehrstein

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Maximilian T. Strauss

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Heinrich Grabmayr

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

  • Peng Yin

    (Wyss Institute for Biologically Inspired Engineering
    Harvard University)

  • Ralf Jungmann

    (Ludwig Maximilian University
    Max Planck Institute of Biochemistry)

Abstract

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.

Suggested Citation

  • Florian Schueder & Juanita Lara-Gutiérrez & Brian J. Beliveau & Sinem K. Saka & Hiroshi M. Sasaki & Johannes B. Woehrstein & Maximilian T. Strauss & Heinrich Grabmayr & Peng Yin & Ralf Jungmann, 2017. "Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT," Nature Communications, Nature, vol. 8(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-02028-8
    DOI: 10.1038/s41467-017-02028-8
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