Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction
Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction
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DOI：
10.1038/s41467-019-12165-x
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发表时间：
2019-09-20
影响因子：
16.6
通讯作者：
Mueller, Marcel
中科院分区：
文献类型：
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作者：
Markwirth, Andreas;Lachetta, Mario;Mueller, Marcel
超分辨结构照明显微镜（SR-SIM）是能够超越光学衍射极限的最快荧光显微镜技术之一。目前的定制仪器能够以高采集速度提供两倍的分辨率增强。SR-SIM通常是一个两步过程，原始数据采集和随后耗时的图像重建后处理。相比之下，宽场和（多点）共焦技术立即产生高分辨率图像。SR-SIM通过将支持视频速率的SIM与快速重建软件紧密集成，也可以实现这种即时性。在这里，我们介绍了VIGOR的即时SR-SIM（视频速率即时GPU加速开源重建）。我们演示了视频帧速率下的多色SR-SIM，测量和重建图像显示之间的延迟小于250 ms。这是通过使用新的、GPU增强的、支持网络的图像重建软件修改和扩展高速SR-SIM图像采集来实现的。我们展示了多种颜色的生物样品的高速测量和移动线粒体的实时成像，作为细胞内动力学的一个例子。
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.