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Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction

Author

Listed:
  • Andreas Markwirth

    (Bielefeld University)

  • Mario Lachetta

    (Bielefeld University)

  • Viola Mönkemöller

    (Bielefeld University
    KU Leuven)

  • Rainer Heintzmann

    (Leibniz Institute of Photonic Technology
    Friedrich-Schiller-University Jena)

  • Wolfgang Hübner

    (Bielefeld University)

  • Thomas Huser

    (Bielefeld University)

  • Marcel Müller

    (Bielefeld University
    KU Leuven)

Abstract

Super-resolved structured illumination microscopy (SR-SIM) is among the fastest fluorescence microscopy techniques capable of surpassing the optical diffraction limit. Current custom-build instruments are able to deliver two-fold resolution enhancement with high acquisition speed. SR-SIM is usually a two-step process, with raw-data acquisition and subsequent, time-consuming post-processing for image reconstruction. In contrast, wide-field and (multi-spot) confocal techniques produce high-resolution images instantly. Such immediacy is also possible with SR-SIM, by tight integration of a video-rate capable SIM with fast reconstruction software. Here we present instant SR-SIM by VIGOR (Video-rate Immediate GPU-accelerated Open-Source Reconstruction). We demonstrate multi-color SR-SIM at video frame-rates, with less than 250 ms delay between measurement and reconstructed image display. This is achieved by modifying and extending high-speed SR-SIM image acquisition with a new, GPU-enhanced, network-enabled image-reconstruction software. We demonstrate high-speed surveying of biological samples in multiple colors and live imaging of moving mitochondria as an example of intracellular dynamics.

Suggested Citation

  • Andreas Markwirth & Mario Lachetta & Viola Mönkemöller & Rainer Heintzmann & Wolfgang Hübner & Thomas Huser & Marcel Müller, 2019. "Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-12165-x
    DOI: 10.1038/s41467-019-12165-x
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