Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan.
Research Center for Applied Sciences, Academia Sinica, Taipei, Taiwan.
J Biomed Opt. 2021 Nov;26(11). doi: 10.1117/1.JBO.26.11.116503.
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.
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.
Axicon imaging is achieved by imposing an axicon lens in the detection part of LSFM.
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.
We demonstrate the fast three-dimensional imaging capability of 2p-LSFM with axicon imaging by recording the rapid dynamics of physiological processes.
双光子显微镜已成为用于深层组织荧光成像的标准平台。然而，传统双光子显微镜中的点扫描限制了体积图像采集的速度。
为了获得快速和深层的体积图像，我们将双光子光片荧光显微镜（2p-LSFM）与轴棱锥成像相结合，在 2p-LSFM 中获得了扩展的景深（DOF）。
通过在 LSFM 的检测部分施加轴棱锥透镜来实现轴棱锥成像。
轴棱锥成像的景深比传统成像透镜扩展了 20 多倍，释放了 LSFM 中的同步扫描。我们以高达 30 Hz 的速率捕获了斑马鱼幼虫中动态跳动的心脏和红细胞的图像。
我们通过记录生理过程的快速动态来展示带轴棱锥成像的 2p-LSFM 的快速三维成像能力。
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