组织器官透明化技术在三维成像研究中的应用

Application of clearing methods on tissues in the three-dimensional imaging research

随着显微成像技术的发展和大数据采集处理系统的不断革新,生物组织成像分析由二维组织切片成像发展到组织器官的三维成像技术。但是,由于组织器官的不透明导致的光线散射,使成像的深度成为一大难题。近年来,组织器官透明化技术取得了很大的发展,该技术通过提高组织的透明度,大大加深了成像深度,从而使得对组织器官的三维成像分析在不需要进行组织切片的情况下得以实现。根据透明化试剂亲水性不同,可将其分为脂溶性透明化技术和水溶性透明化技术,前者减少了组织中的水分,使透明化更加彻底有效;而后者则利用水分子在蛋白分子周围形成的水化膜提高了荧光蛋白稳定性,更利于研究的进行。我们将按照分类对不同透明化技术作出介绍,并从透明化时间长短、荧光保留度等方面进行优劣比较。

With the development of the microscopy and the big data acquisition processing system, the research direction gradually becomes the construction of biological structures by 3D imaging. However, because of opaque which leads to scattering of imaging light through the tissues, the depth of the imaging becomes a challenge. A technique named ＂clearing＂ can increase the organization transparency so that they can enhance the depth of the imaging and have a three- dimensional reconstruction. Being hydrophilic, the clearing reagents were divided into solvent-based clearing and aqueous- based clearing. The solvent-based clearing reduced water in the tissue and aqueous-based clearing used the hydration shell made by water to keep the fluorescent protein stability. In this article we introduced different clearing method and compared them from clearing time and retention in the fluorescence.