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Nat Commun. 2019 Sep 20;10(1):4315. doi: 10.1038/s41467-019-12165-x.

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

Author information

1
Biomolecular Photonics, Faculty of Physics, Bielefeld University, Bielefeld, Germany.
2
Laboratory for NanoBiology, Department of Chemistry, KU Leuven, Leuven, Belgium.
3
Leibniz Institute of Photonic Technology, Jena, Germany.
4
Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller-University Jena, Jena, Germany.
5
Biomolecular Photonics, Faculty of Physics, Bielefeld University, Bielefeld, Germany. thomas.huser@physik.uni-bielefeld.de.
6
Biomolecular Photonics, Faculty of Physics, Bielefeld University, Bielefeld, Germany. muellerphysics@gmail.com.
7
Laboratory for NanoBiology, Department of Chemistry, KU Leuven, Leuven, Belgium. muellerphysics@gmail.com.

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.

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