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Super-resolution imaging in whole cells and tissues via DNA-PAINT on a spinning disk confocal with optical photon reassignment

Author

Listed:
  • Cecilia Zaza

    (University College London)

  • Megan D. Joseph

    (University College London)

  • Olivia P. L. Dalby

    (University College London
    University College London)

  • Rhian F. Walther

    (University College London)

  • Karol Kołątaj

    (University of Fribourg
    University of Fribourg)

  • Germán Chiarelli

    (University of Fribourg)

  • Franck Pichaud

    (University College London
    University College London)

  • Guillermo P. Acuna

    (University of Fribourg
    University of Fribourg)

  • Sabrina Simoncelli

    (University College London
    University College London)

Abstract

Single-Molecule Localization Microscopy (SMLM) has traditionally faced challenges to optimize signal-to-noise ratio, penetration depth, field-of-view (FOV), and spatial resolution simultaneously. Here, we show that DNA-PAINT imaging on a Spinning Disk Confocal with Optical Photon Reassignment (SDC-OPR) system overcomes these trade-offs, enabling high-resolution imaging across multiple cellular layers and large FOVs. We demonstrate the system’s capability with DNA origami constructs and biological samples, including nuclear pore complexes, mitochondria, and microtubules, achieving a spatial resolution of 6 nm in the basal plane and sub-10 nm localization precision at depths of 9 µm within a 53 × 53 µm² FOV. Additionally, imaging of the developing Drosophila eye epithelium at depths up to 9 µm with sub-13 nm average localization precision, reveals distinct E-cadherin populations in adherens junctions. Quantitative analysis of Collagen IV deposition in this epithelium indicated an average of 46 ± 27 molecules per secretory vesicle. These results underscore the versatility of DNA-PAINT on an SDC-OPR for advancing super-resolution imaging in complex biological systems.

Suggested Citation

  • Cecilia Zaza & Megan D. Joseph & Olivia P. L. Dalby & Rhian F. Walther & Karol Kołątaj & Germán Chiarelli & Franck Pichaud & Guillermo P. Acuna & Sabrina Simoncelli, 2025. "Super-resolution imaging in whole cells and tissues via DNA-PAINT on a spinning disk confocal with optical photon reassignment," Nature Communications, Nature, vol. 16(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-60263-w
    DOI: 10.1038/s41467-025-60263-w
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    References listed on IDEAS

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    1. Matthieu Cavey & Matteo Rauzi & Pierre-François Lenne & Thomas Lecuit, 2008. "A two-tiered mechanism for stabilization and immobilization of E-cadherin," Nature, Nature, vol. 453(7196), pages 751-756, June.
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