Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction
| Autor: | Markwirth, Andreas, Lachetta, Mario, Mönkemöller, Viola, Heintzmann, Rainer, Hübner, Wolfgang, Huser, Thomas, Müller, Marcel |
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| Rok vydání: | 2019 |
| Předmět: |
SR-SIM
Science Biophysics ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION Color Article Fluorescence imaging Imaging Imaging Three-Dimensional Image Processing Computer-Assisted Humans lcsh:Science Lighting ComputingMethodologies_COMPUTERGRAPHICS VIGOR Microscopy Video Nanoscale biophysics Lasers Optical Imaging Imaging and sensing Mitochondria Microscopy Fluorescence microscopy lcsh:Q Software |
| Zdroj: | Nature Communications Nature Communications, Vol 10, Iss 1, Pp 1-11 (2019) |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-019-12165-x |
| Popis: | 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. Sequential acquisition and image reconstruction in super-resolved structured illumination microscopy (SR-SIM) is time-consuming. Here the authors optimise both acquisition and reconstruction software to achieve multicolour SR-SIM at video frame-rates with reconstructed images displaying with only milliseconds delay during the experiment. |
| Databáze: | OpenAIRE |
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