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

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Date
2019
Volume
10
Issue
Journal
Nature Communications
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Publisher
Berlin : Nature Publishing
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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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 (Berlin : Nature Publishing). Berlin : Nature Publishing. https://doi.org//10.1038/s41467-019-12165-x
License
CC BY 4.0 Unported