ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging

作者:Lee Eunsoo; Choi Jungyoon; Jo Youhwa; Kim Joo Yeon; Jang Yu Jin; Lee Hye Myeong; Kim So Yeun; Lee Ho Jae; Cho Keunchang; Jung Neoncheol; Hur Eun Mi; Jeong Sung Jin; Moon Cheil; Choe Youngshik; Rhyu Im Joo; Kim Hyun; Sun Woong
来源:Scientific Reports, 2016, 6(1): 18631.
DOI:10.1038/srep18631

摘要

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.

  • 出版日期2016-1-11