Choi Jungyoon, Lee Eunsoo, Kim June Hoan, Sun Woong
Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Seoul 02841, Korea.
Exp Neurobiol. 2019 Jun;28(3):436-445. doi: 10.5607/en.2019.28.3.436. Epub 2019 Jun 26.
Over the last two decades, several tissue clearing methodologies have been established that render tissues optically transparent and allow imaging of unsectioned tissues of significant volumes, thus improving the capacity to study the relationships between cell and 3D tissue architecture. Despite these technical advances, the important unsolved challenges that these methods face include complexity, time, consistency of tissue size before and after clearing, and ability to immunolabel various antibodies in cleared tissue. Here, we established very simple and fast tissue clearing protocol, FxClear, which involves acrylamide-free electrophoretic tissue clearing (ETC). By removal of the acrylamide infusion step, we were able to achieve fast reaction time, smaller tissue expansion, and higher immunoreactivity. Especially, immunoreactivity and fluorescence intensity were increased in FxClear-processed tissues compared to un-cleared tissues. Our protocol may be suitable for small-sized biopsy samples for 3D pathological examinations.
在过去二十年中，已经建立了几种组织透明化方法，这些方法使组织具有光学透明性，并允许对大量未切片的组织进行成像，从而提高了研究细胞与三维组织结构之间关系的能力。尽管有这些技术进步，但这些方法面临的重要未解决挑战包括复杂性、时间、透明化前后组织大小的一致性，以及在透明化组织中对各种抗体进行免疫标记的能力。在这里，我们建立了一种非常简单快速的组织透明化方案FxClear，它涉及无丙烯酰胺的电泳组织透明化（ETC）。通过去除丙烯酰胺灌注步骤，我们能够实现快速反应时间、较小的组织膨胀和更高的免疫反应性。特别是，与未处理的组织相比，FxClear处理的组织中免疫反应性和荧光强度有所增加。我们的方案可能适用于用于三维病理检查的小尺寸活检样本。
