Image quality guided smart rotation improves coverage in microscopy
Image quality guided smart rotation improves coverage in microscopy
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DOI：
10.1038/s41467-019-13821-y
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发表时间：
2020-01-09
影响因子：
16.6
通讯作者：
Huisken, Jan
中科院分区：
文献类型：
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作者：
He, Jiaye;Huisken, Jan
荧光显微镜是生物发现的重要工具。人们不断需要在更大的视野范围内获得更好的空间分辨率。尽管在提高光学仪器的理论分辨率和速度方面已经取得了长足的进步，但在介观样品中，图像质量仍然在很大程度上受到样品光学特性的限制。在选择性平面照明显微镜 (SPIM) 中，可实现的光学性能因照明和检测中遇到的光学退化而受到阻碍。通过样本旋转或附加光路进行多视图成像是提高样本覆盖率的流行策略。在这项工作中，我们引入了一种智能旋转工作流程，该工作流程利用动态图像分析来识别最佳光片成像方向。智能旋转工作流程优于传统方法，无需额外硬件，并使用相同或更少的角度实现更好的样品覆盖，从而减少数据量和光毒性。
Fluorescence microscopy is an essential tool for biological discoveries. There is a constant demand for better spatial resolution across a larger field of view. Although strides have been made to improve the theoretical resolution and speed of the optical instruments, in mesoscopic samples, image quality is still largely limited by the optical properties of the sample. In Selective Plane Illumination Microscopy (SPIM), the achievable optical performance is hampered by optical degradations encountered in both the illumination and detection. Multi-view imaging, either through sample rotation or additional optical paths, is a popular strategy to improve sample coverage. In this work, we introduce a smart rotation workflow that utilizes on-the-fly image analysis to identify the optimal light sheet imaging orientations. The smart rotation workflow outperforms the conventional approach without additional hardware and achieves a better sample coverage using the same number of angles or less and thereby reduces data volume and phototoxicity.