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Smart scanning for low-illumination and fast RESOLFT nanoscopy in vivo

Author

Listed:
  • Jes Dreier

    (KTH Royal Institute of Technology)

  • Marco Castello

    (Istituto Italiano di Tecnologia)

  • Giovanna Coceano

    (KTH Royal Institute of Technology)

  • Rodrigo Cáceres

    (PSL Research University, CNRS
    Sorbonne Université
    Université Paris Descartes)

  • Julie Plastino

    (PSL Research University, CNRS
    Sorbonne Université)

  • Giuseppe Vicidomini

    (Istituto Italiano di Tecnologia)

  • Ilaria Testa

    (KTH Royal Institute of Technology)

Abstract

RESOLFT fluorescence nanoscopy can nowadays image details far beyond the diffraction limit. However, signal to noise ratio (SNR) and temporal resolution are still a concern, especially deep inside living cells and organisms. In this work, we developed a non-deterministic scanning approach based on a real-time feedback system which speeds up the acquisition up to 6-fold and decreases the light dose by 70–90% for in vivo imaging. Also, we extended the information content of the images by acquiring the complete temporal evolution of the fluorescence generated by reversible switchable fluorescent proteins. This generates a series of images with different spatial resolution and SNR, from conventional to RESOLFT images, which combined through a multi-image deconvolution algorithm further enhances the effective resolution. We reported nanoscale imaging of organelles up to 35 Hz and actin dynamics during an invasion process at a depth of 20–30 µm inside a living Caenorhabditis elegans worm.

Suggested Citation

  • Jes Dreier & Marco Castello & Giovanna Coceano & Rodrigo Cáceres & Julie Plastino & Giuseppe Vicidomini & Ilaria Testa, 2019. "Smart scanning for low-illumination and fast RESOLFT nanoscopy in vivo," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-08442-4
    DOI: 10.1038/s41467-019-08442-4
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