Significance: Two-photon microscopy has become the standard platform for deep-tissue fluorescence imaging. However, the use of point scanning in conventional two-photon microscopy limits the speed of volumetric image acquisition. Aim: To obtain fast and deep volumetric images, we combine two-photon light sheet fluorescence microscopy (2p-LSFM) and axicon imaging that yields an extended depth of field (DOF) in 2p-LSFM. Approach: Axicon imaging is achieved by imposing an axicon lens in the detection part of LSFM. Results: The DOF with axicon imaging is extended more than 20-fold over that of a conventional imaging lens, liberating the synchronized scanning in LSFM. We captured images of dynamic beating hearts and red blood cells in zebrafish larvae at volume acquisition rates up to 30 Hz. Conclusions: We demonstrate the fast three-dimensional imaging capability of 2p-LSFM with axicon imaging by recording the rapid dynamics of physiological processes.

中文翻译：

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具有轴锥成像的双光子扫描光片荧光显微镜，用于快速体积成像

意义：双光子显微镜已成为深部组织荧光成像的标准平台。然而，在传统的双光子显微镜中使用点扫描限制了体积图像采集的速度。目标：为了获得快速和深的体积图像，我们结合了双光子光片荧光显微镜 (2p-LSFM) 和轴锥成像，在 2p-LSFM 中产生了扩展的景深 (DOF)。方法：轴锥成像是通过在 LSFM 的检测部分施加轴锥透镜来实现的。结果：轴像成像的自由度是传统成像镜头的 20 倍以上，解放了 LSFM 中的同步扫描。我们以高达 30 Hz 的体积采集率捕获了斑马鱼幼虫中动态跳动的心脏和红细胞的图像。结论：