High-contrast, synchronous volumetric imaging with selective volume illumination microscopy
High-contrast, synchronous volumetric imaging with selective volume illumination microscopy
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DOI：
10.1038/s42003-020-0787-6
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发表时间：
2020-02-14
影响因子：
5.9
通讯作者：
Fraser, Scott E.
中科院分区：
文献类型：
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作者：
Truong, Thai, V;Holland, Daniel B.;Fraser, Scott E.
光场荧光显微镜独特地通过在一个快照中捕获扩展的体积来提供快速、同步的体积成像，但是由于其宽场照明策略产生的背景信号而经常遭受低对比度。我们实现了基于光场的选择性体积照明显微镜（SVIM），其中照明仅限于感兴趣的体积，去除从无关的样品体积产生的背景，并显着提高图像的对比度。我们通过捕获海水中流动细菌的细胞分辨率3D电影来展示SVIM的能力，因为它们在鱿鱼共生伙伴中定殖，以及在斑马鱼幼虫中跳动的心脏和全脑神经活动。这些应用展示了我们设想的SVIM将能够实现的成像应用的广度，在以单细胞分辨率、快速体积速率和高对比度捕获组织尺度3D动态生物系统以揭示潜在的生物学。一种结合光场显微术与选择性平面照明显微术以提高影像对比度与解析度的方法。他们通过将SVIM应用于海水中流动的细菌以及斑马鱼幼鱼跳动的心脏和整个大脑来生成细胞分辨率的3D电影。
Light-field fluorescence microscopy uniquely provides fast, synchronous volumetric imaging by capturing an extended volume in one snapshot, but often suffers from low contrast due to the background signal generated by its wide-field illumination strategy. We implemented light-field-based selective volume illumination microscopy (SVIM), where illumination is confined to only the volume of interest, removing the background generated from the extraneous sample volume, and dramatically enhancing the image contrast. We demonstrate the capabilities of SVIM by capturing cellular-resolution 3D movies of flowing bacteria in seawater as they colonize their squid symbiotic partner, as well as of the beating heart and brain-wide neural activity in larval zebrafish. These applications demonstrate the breadth of imaging applications that we envision SVIM will enable, in capturing tissue-scale 3D dynamic biological systems at single-cell resolution, fast volumetric rates, and high contrast to reveal the underlying biology.Thai Truong et al. present light-field-based selective volume illumination microscopy (SVIM), a method for enhancing image contrast and resolution by combining light-field microscopy and selective plane illumination microscopy. They generate cellular-resolution 3D movies by applying SVIM to flowing bacteria in seawater and to the beating heart and whole brain of larval zebrafish.