Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577, Japan.
Sci Rep. 2019 Aug 12;9(1):11687. doi: 10.1038/s41598-019-48265-3.
The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a method enabling axially resolved volumetric imaging without a moving observation plane in the framework of laser scanning microscopy. A scanning light needle spot with an extended focal depth provides excitation, which normally produces a deep focus image with a loss of depth information. In our method, the depth information is retrieved from transposed lateral information on an array detector by utilising non-diffracting and self-bending characteristics imposed on fluorescent signals. This technique, implemented in two-photon microscopy, achieves truly volumetric images constructed from a single raster scan of a light needle, which has the capability to significantly reduce the acquisition time.
在包括生命科学在内的各个领域，对快速三维容积成像的需求正在增加。激光扫描荧光显微镜已被广泛用于此目的；然而，体积图像是通过具有变化的观察平面的二维图像堆叠构建的，最终限制了采集速度。在这里，我们提出了一种在激光扫描显微镜框架内无需移动观察平面即可实现轴向分辨体积成像的方法。扩展焦深的扫描光针点提供激发，通常会产生深度信息丢失的深焦图像。在我们的方法中，深度信息是通过利用施加在荧光信号上的无衍射和自弯曲特性从阵列探测器上的转置横向信息中检索出来的。这项在双光子显微镜中实现的技术，从单个光针的单次光栅扫描构建真正的体积图像，这有可能显著缩短采集时间。
