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

Andreas Markwirth, Mario Lachetta, Viola Mönkemöller, Rainer Heintzmann, Wolfgang Hübner, Thomas Huser () and Marcel Müller ()
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Andreas Markwirth: Bielefeld University
Mario Lachetta: Bielefeld University
Viola Mönkemöller: Bielefeld University
Rainer Heintzmann: Leibniz Institute of Photonic Technology
Wolfgang Hübner: Bielefeld University
Thomas Huser: Bielefeld University
Marcel Müller: Bielefeld University

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-12165-x

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