Single-shot 3D wide-field fluorescence imaging with a Computational Miniature Mesoscope.
Single-shot 3D wide-field fluorescence imaging with a Computational Miniature Mesoscope.
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DOI：
10.1126/sciadv.abb7508
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发表时间：
2020-10
期刊：
影响因子：
13.6
通讯作者：
Tian L
中科院分区：
文献类型：
--
作者：
Xue Y;Davison IG;Boas DA;Tian L
计算微型显微镜能够实现单次大规模3D荧光成像，具有单细胞水平的分辨率。荧光显微镜是生物学和神经科学不可或缺的。对自由行为动物进行记录的需求进一步推动了小型化显微镜（微型镜）的发展。然而，传统的显微镜/微型镜固有地受到其有限的空间带宽积、浅景深（DOF）以及无法分辨三维（3D）分布式发射器的限制。在这里，我们提出了一个计算微型显微镜（CM 2），克服了这些瓶颈，并使单次拍摄的3D成像跨越8 mm × 7 mm的视场和2.5 mm的自由度，实现7 μm的横向分辨率和优于200 μm的轴向分辨率。CM 2采用紧凑的轻量化设计，集成了用于成像的微透镜阵列和用于激发的发光二极管阵列。其扩展的成像能力是通过计算成像来实现的，计算成像通过算法来增强光学器件。我们在实验上验证了三维荧光样品的介观成像能力。我们进一步量化了散射和背景荧光对幻影实验的影响。
Computational Miniature Mesoscope enables single-shot large-scale 3D fluorescence imaging with single-cell level resolution. Fluorescence microscopes are indispensable to biology and neuroscience. The need for recording in freely behaving animals has further driven the development in miniaturized microscopes (miniscopes). However, conventional microscopes/miniscopes are inherently constrained by their limited space-bandwidth product, shallow depth of field (DOF), and inability to resolve three-dimensional (3D) distributed emitters. Here, we present a Computational Miniature Mesoscope (CM2) that overcomes these bottlenecks and enables single-shot 3D imaging across an 8 mm by 7 mm field of view and 2.5-mm DOF, achieving 7-μm lateral resolution and better than 200-μm axial resolution. The CM2 features a compact lightweight design that integrates a microlens array for imaging and a light-emitting diode array for excitation. Its expanded imaging capability is enabled by computational imaging that augments the optics by algorithms. We experimentally validate the mesoscopic imaging capability on 3D fluorescent samples. We further quantify the effects of scattering and background fluorescence on phantom experiments.