Zhou Can, Zheng Ting, Luo Ting, Yan Cheng, Sun Qingtao, Ren Miao, Zhao Peilin, Chen Wu, Ji Bingqing, Wang Zhi, Li Anan, Gong Hui, Li Xiangning
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan 430074, China.
These authors contributed equally.
Biomed Opt Express. 2020 Nov 12;11(12):7132-7149. doi: 10.1364/BOE.405801. eCollection 2020 Dec 1.
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.
光学透明化方法在全器官三维生物信息获取中被广泛应用。然而，透明化组织的成像质量常常受到组织透明化不均匀的限制。通过将组织透明化与基于机械切片的全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，从而在浅层和深层都获得各向同性和高分辨率的图像。然而，对于光学透明化的生物组织来说，保持良好的切片性能仍然具有挑战性。在此，我们开发了一种名为M-CUBIC（可加工CUBIC）的透明化方法，该方法将改良的CUBIC方法与PNAGA（聚N-丙烯酰甘氨酰胺）水凝胶包埋相结合，使组织透明化的同时提高其切片性能。利用高通量光片断层扫描平台（HLTP）和荧光显微光学切片断层扫描（fMOST），我们从完整的小鼠大脑中获取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。该方法可用于获取多种类型生物器官具有亚细胞分辨率的微观结构，有助于生物学研究。
