A versatile, multi-laser twin-microscope system for light-sheet imaging
A versatile, multi-laser twin-microscope system for light-sheet imaging
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DOI：
10.1063/1.5144487
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发表时间：
2020-05-01
影响因子：
1.6
通讯作者：
Truong, Thai V.
中科院分区：
文献类型：
--
作者：
Keomanee-Dizon, Kevin;Fraser, Scott E.;Truong, Thai V.
与传统的点扫描显微镜相比，光片显微镜可提供更快的成像速度并降低光毒性，使其成为持续数小时或数天的生物动力学成像的首选技术。这种延长的成像时间带来了挑战，因为它减少了一天内可以成像的标本数量。在这里，我们提出了一种多功能光片成像仪器，它结合了两个独立控制的显微镜双胞胎，使它们可以共享超快近红外激光器和一组连续波可见激光器，从而提高吞吐量并降低成本。为了允许对各种样本进行成像，每个显微镜双体都提供灵活的成像参数，包括（i）在单光子和/或双光子激发模式下操作，（ii）通过三个正交激发臂传输一到三个光片，（iii）亚微米到微米成像分辨率，（iv）多色兼容性，以及（v）直立（提供倒置）检测几何结构。我们提供双显微镜设计的详细描述，以帮助希望构建和使用类似系统的仪器制造商。我们通过对完整斑马鱼胚胎中跳动的心脏进行快速成像、对厚厚的患者来源肿瘤类器官进行深度成像以及对行为幼虫斑马鱼的神经活动进行温和的全脑成像，展示了该仪器在生物学研究中的多功能性。
Light-sheet microscopy offers faster imaging and reduced phototoxicity in comparison to conventional point-scanning microscopy, making it a preferred technique for imaging biological dynamics for durations of hours or days. Such extended imaging sessions pose a challenge, as it reduces the number of specimens that can be imaged in a given day. Here, we present a versatile light-sheet imaging instrument that combines two independently controlled microscope-twins, built so that they can share an ultrafast near-infrared laser and a bank of continuous-wave visible lasers, increasing the throughput and decreasing the cost. To permit a wide variety of specimens to be imaged, each microscope-twin provides flexible imaging parameters, including (i) operation in one-photon and/or two-photon excitation modes, (ii) delivery of one to three light-sheets via a trio of orthogonal excitation arms, (iii) sub-micron to micron imaging resolution, (iv) multicolor compatibility, and (v) upright (with provision for inverted) detection geometry. We offer a detailed description of the twin-microscope design to aid instrument builders who wish to construct and use similar systems. We demonstrate the instrument's versatility for biological investigation by performing fast imaging of the beating heart in an intact zebrafish embryo, deep imaging of thick patient-derived tumor organoids, and gentle whole-brain imaging of neural activity in behaving larval zebrafish.