Opt Lett. 2018 Nov 1;43(21):5271-5274. doi: 10.1364/OL.43.005271.
Optical sectioning has become an indispensable technique for high-speed volumetric imaging in the past decade. Here we present a novel optical-sectioning method that produces a thin plane of illumination by exploiting the spatial and temporal properties of multiphoton excitation. Critically, the illumination and detection share the same optical path, as in a conventional epi-fluorescence microscope configuration. Therefore, the imaged sample can be prepared as for standard fluorescence microscopy. Our method also leads to a laterally structured illumination pattern, and this feature can be utilized in structured illumination microscopy to further enhance the imaging performance. We show an example of such an approach, which achieves axial resolution finer than confocal microscopy. We also demonstrate the potential of the new method for biological applications by performing three-dimensional imaging of living Caenorhabditis elegans.
在过去的十年中，光学切片已成为高速体积成像不可或缺的技术。在这里，我们提出了一种新的光学切片方法，该方法利用多光子激发的空间和时间特性产生薄的照明平面。关键是，照明和检测共享相同的光路，就像传统的 epi-荧光显微镜配置一样。因此，可以像进行标准荧光显微镜一样准备成像样品。我们的方法还导致了横向结构的照明图案，并且该特征可用于结构照明显微镜中以进一步提高成像性能。我们展示了这样一种方法的示例，其实现了优于共聚焦显微镜的轴分辨率。我们还通过对活体秀丽隐杆线虫进行三维成像，展示了新方法在生物学应用中的潜力。
