Optical clearing methods are widely used for three-dimensional biological information acquisition in the whole organ. However, the imaging quality of cleared tissues is often limited by ununiformed tissue clearing. By combining tissue clearing with mechanical sectioning based whole organ imaging system, we can reduce the influence of light scattering and absorption on the tissue to get isotropic and high resolution in both superficial and deep layers. However, it remains challenging for optical cleared biological tissue to maintain good sectioning property. Here, we developed a clearing method named M-CUBIC (machinable CUBIC), which combined a modified CUBIC method with PNAGA (poly-N-acryloyl glycinamide) hydrogel embedding to transparentize tissue while improving its sectioning property. With high-throughput light-sheet tomography platform (HLTP) and fluorescent micro-optical sectioning tomography (fMOST), we acquired continuous datasets with subcellular resolution from intact mouse brains for single neuron tracing, as well as the fine vascular structure of kidneys. This method can be used to acquire microstructures of multiple types of biological organs with subcellular resolutions, which can facilitate biological research.

中文翻译：

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可加工的光学清除方法以亚细胞分辨率连续成像大体积组织

光学清除方法被广泛用于整个器官的三维生物信息获取。但是，清除的组织的成像质量通常受组织清除不均匀的限制。通过将组织清除与基于机械切片的整个器官成像系统相结合，我们可以减少光散射和吸收对组织的影响，从而在表层和深层均获得各向同性和高分辨率。然而，光学清除的生物组织保持良好的切片性能仍然具有挑战性。在这里，我们开发了一种称为M-CUBIC（可加工CUBIC）的清除方法，该方法将改进的CUBIC方法与PNAGA（聚N-丙烯酰甘氨酰胺）水凝胶嵌入相结合，以使组织透明化，同时提高其切片性能。利用高通量光片断层扫描平台（HLTP）和荧光微光学断层扫描（fMOST），我们从完整的小鼠大脑获得了具有亚细胞分辨率的连续数据集，用于单个神经元追踪以及肾脏的精细血管结构。该方法可用于获取具有亚细胞分辨率的多种类型生物器官的微观结构，从而有助于生物学研究。