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

Florian Schueder, Juanita Lara-Gutiérrez, Brian J. Beliveau, Sinem K. Saka, Hiroshi M. Sasaki, Johannes B. Woehrstein, Maximilian T. Strauss, Heinrich Grabmayr, Peng Yin () and Ralf Jungmann ()
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Florian Schueder: Ludwig Maximilian University
Juanita Lara-Gutiérrez: Wyss Institute for Biologically Inspired Engineering
Brian J. Beliveau: Wyss Institute for Biologically Inspired Engineering
Sinem K. Saka: Wyss Institute for Biologically Inspired Engineering
Hiroshi M. Sasaki: Wyss Institute for Biologically Inspired Engineering
Johannes B. Woehrstein: Ludwig Maximilian University
Maximilian T. Strauss: Ludwig Maximilian University
Heinrich Grabmayr: Ludwig Maximilian University
Peng Yin: Wyss Institute for Biologically Inspired Engineering
Ralf Jungmann: Ludwig Maximilian University

Nature Communications, 2017, vol. 8, issue 1, 1-9

Abstract: 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.

Date: 2017
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DOI: 10.1038/s41467-017-02028-8

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