ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging.
ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging.
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DOI：
10.1038/srep18631
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发表时间：
2016-01-11
影响因子：
4.6
通讯作者：
Sun W
中科院分区：
文献类型：
--
作者：
Lee E;Choi J;Jo Y;Kim JY;Jang YJ;Lee HM;Kim SY;Lee HJ;Cho K;Jung N;Hur EM;Jeong SJ;Moon C;Choe Y;Rhyu IJ;Kim H;Sun W
在细胞水平上理解器官和生物体的结构组织是生物学中的一个基本挑战。这一任务已经通过从连续切片的组织中获取的图像重建三维结构来实现，这不仅是劳动密集型和耗时的，而且容易出错。组织清除技术的最新进展允许可视化未切片的整个组织或整个身体内的细胞结构和神经网络。然而，目前可用的协议需要长的处理时间。在这里，我们提出了快速和高度可重复的ACT-PRESTO（主动澄清技术-压力相关的大分子进入器官的有效和稳定转移）方法，该方法在1天内清除组织或全身，同时保留组织结构和源自内源性荧光蛋白的基于蛋白质的信号。此外，ACT-PRESTO与传统的免疫标记方法兼容，并通过施加压力加速抗体渗透到厚标本中。该方法的速度和一致性将允许对完整器官和身体中的正常和病理特征进行高内容的映射和分析。
Understanding the structural organization of organs and organisms at the cellular level is a fundamental challenge in biology. This task has been approached by reconstructing three-dimensional structure from images taken from serially sectioned tissues, which is not only labor-intensive and time-consuming but also error-prone. Recent advances in tissue clearing techniques allow visualization of cellular structures and neural networks inside of unsectioned whole tissues or the entire body. However, currently available protocols require long process times. Here, we present the rapid and highly reproducible ACT-PRESTO (active clarity technique-pressure related efficient and stable transfer of macromolecules into organs) method that clears tissues or the whole body within 1 day while preserving tissue architecture and protein-based signals derived from endogenous fluorescent proteins. Moreover, ACT-PRESTO is compatible with conventional immunolabeling methods and expedites antibody penetration into thick specimens by applying pressure. The speed and consistency of this method will allow high-content mapping and analysis of normal and pathological features in intact organs and bodies.