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

Andreas Markwirth, Mario Lachetta, Viola Mönkemöller, Rainer Heintzmann, Wolfgang Hübner, Thomas Huser, Marcel Müller

Research output: Contribution to journalArticle

5 Scopus citations

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.

Original languageEnglish (US)
Number of pages1
JournalNature communications
Volume10
Issue number1
DOIs
StatePublished - Sep 20 2019
Externally publishedYes

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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    Markwirth, A., Lachetta, M., Mönkemöller, V., Heintzmann, R., Hübner, W., Huser, T., & Müller, M. (2019). Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction. Nature communications, 10(1). https://doi.org/10.1038/s41467-019-12165-x