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11MACS: RAPID AQUEOUS CLEARING SYSTEM FOR THREE-DIMENSIONAL MAPPING OF
INTACT ORGANS
JINGTAN ZHU, TINGTING YU, YUSHA LI, JIANYI XU, YISONG QI, YINGTAO YAO, YILIN MA, PENG WAN, ZHILONG CHEN, XIANGNING LI, HUI GONG, QINGMING LUO, DAN ZHU*
1Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science
and Technology, China
2
MoE Key Laboratory for Biomedical Photonics, Huazhong University of Science and Technology, Wuhan 430074, China
Email: [email protected]
Abstract
Tissue optical clearing techniques have provided important tools for large-volume imaging. Aqueous-based clearing methods are known for good fluorescence preservation and scalable size maintenance, but are limited by long incubation time, or insufficient clearing performance, or requirements for specialized devices. Additionally, few clearing methods are compatible with widely-used lipophilic dyes while maintaining high clearing performance. Here, to address these issues, m-xylylenediamine (MXDA) is firstly introduced into tissue clearing and used to develop a rapid, highly efficient aqueous clearing method with robust lipophilic dyes compatibility, termed MXDA-based Aqueous Clearing System (MACS). MACS can render whole adult brains highly transparent within 2.5 d and is also applicable for other intact organs. Meanwhile, MACS possesses ideal compatibilitywith multiple probes, especially for lipophilic dyes. MACS achieves three-dimensional (3D) imaging of the intact neural structures labelled by various techniques. Combining MACS with DiI labelling, MACS allows reconstruction of the detailed vascular structures of various organs and generates 3D pathology of glomeruli tufts in healthy and diabetic kidneys. Therefore, MACS provides a useful method for 3D mapping of intact tissues and is expected to facilitate morphological, physiological and pathological studies of various organs.
Figure 1:Overview of MACS clearing method.
