组织透明化技术的研究与应用

文题释义：组织透明技术：应用水溶性有机溶剂或者亲水性试剂，对固定组织通过浸泡、电泳或灌注等方式进行透化处理，然后应用高折射率介质匹配组织折射率，降低光散射，使组织达到光学透明，进而增加成像深度和图像对比度的技术。光散射：光从一种介质进入另外一种介质时，由于介质成分的变化，光的强度和曲度都会相应改变，当遇到两种均匀介质的分界面时，光会发生反射和折射现象，但当光在不均匀介质中传播时，一部分光线会偏离原始传播路径向四面八方传播，即形成光的散射现象。背景：对生物体进行全身细胞分析是生物医学领域的主要挑战之一。组织透明技术结合光学成像和图像处理技术，能够将整个器官或全身快速透明并进行结构和细胞分析，为在生命科学中应用先进的光学技术提供了一个非常有前景的解决方案。目的：分析组织透明技术原理及过程，总结组织透明技术研究进展，介绍组织透明成像技术，讨论组织透明技术在生物医学研究领域中的应用。方法：第一作者以“tissue clearing technique，tissue optical clearing，whole-body imaging，3D imaging”为关键词，检索PubMed数据库发表的相关文献，初检文献168篇，系统整理、筛选后，对纳入的72篇文献进行分析、总结和讨论。结果与结论：目前组织透明技术主要分为两类，有机溶剂透明技术和亲水性试剂透明技术，组织透明步骤包括：①组织固定；②透化作用；③脱色作用；④折光率匹配。组织透明技术能够使组织快速光学透明，显著提高成像深度和图像对比度，结合显微成像技术如共聚焦显微镜、双光子显微镜和光片显微镜等，能够在单细胞水平实现全身或全器官的高分辨率三维成像，加速了生物体全身细胞分析进程。未来，组织透明技术将会持续发展，并将推动新型透化试剂和透化专用显微镜的研发，从而加强从完整机体系统获取结构和分子信息，有助于对完整生物系统的综合理解。 ORCID: 0000-0003-1647-7922(田婷)中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程

Research and application of tissue optical clearing technique

BACKGROUND:Whole-body cell analysis of organisms is one of the major challenges in biomedicine.Tissue optical clearing technique combined with optical imaging and image processing technique can make the whole organ or the body transparent rapidly for structural and cellular analyses,providing a very promising solution for the application of advanced optical technique in life science.OBJECTIVE:To analyze the principle and process of tissue optical clearing technique,to summarize the research progress of tissue optical clearing technique,to present imaging technology for tissue clearing,and to discuss the application of tissue optical clearing technique in biomedical researches.METHODS:The first author searched relevant literatures in PubMed database with the keywords of“tissue optical clearing technique,tissue optical clearing,whole-body imaging and 3D imaging”.A total of 168 articles were initially retrieved.After sorting and screening systematically,72 literatures were included for analysis,summary and discussion.RESULTS AND CONCLUSION:Current tissue optical clearing protocols are divided into two groups:solvent-based clearing methods and hydrophilic reagent-based clearing methods.Tissue clearing is usually conducted by the following steps:(a)tissue fixation,(b)permeabilization,(c)decolorization,and(d)refractive index matching.Tissue optical clearing technique can rapidly transform tissue into an optically transparent form,improving imaging depth and contrast.Combined with microscope imaging techniques such as confocal,two-photon and light sheet microscopes,tissue clearing can achieve 3D imaging of the whole organ or body at cellular resolution,accelerating the process of whole-body cell analysis of organisms.Tissue optical clearing technique will continue to evolve in the future,promote the development of new clearing reagents and clearing optimized microscopes,further enhance the acquisition of structural and molecular information from intact systems and contribute to the comprehensive understanding of whole biological systems.