A method for ultrafast tissue clearing that preserves fluorescence for multimodal and longitudinal brain imaging

作者：Qing‐Hong Shan, Xin‐Ya Qin, Nan Zhou, Chuan Huang, Yu Wang, Peng Chen, Jiang‐Ning Zhou · 发表于：BMC Biology · 年份：2022 · DOI：10.1186/s12915-022-01275-6 · 被引用次数：21 · 研究领域：Advanced Fluorescence Microscopy Techniques、Optical Coherence Tomography Applications、Cancer Research and Treatments

BACKGROUND: Tissue-clearing techniques have recently been developed to make tissues transparent for three-dimensional (3D) imaging at different scales, including single-cell resolution. However, current tissue-clearing workflows have several disadvantages, including complex protocols, time-consuming application, and fluorescence quenching. Additionally, they can be used mainly for clearing larger-volume samples, preventing wide and easy applicability in conventional experimental approaches. In this study, we aimed to develop a versatile, fast, and convenient method for clearing thin and semi-thick samples, which can be used for three-dimensional imaging of experimental or even clinical samples. RESULTS: We developed an alkaline solution (AKS) containing a combination of 2,2'-thiodiethanol (TDE), DMSO, D-sorbitol, and Tris for tissue clearing, as the alkaline environment is suitable for maintaining the fluorescence of most commonly used fluorescence protein GFP and its variants, and tested its clearing effect on samples from mice and human brains. We assessed the clearing speed, the preservation of fluorescence protein and dyes, and the imaging depth and quality. The results showed that AKS treatment rapidly cleared 300-μm-thick brain slices and 1-mm-thick slices from different organs within 5 min and 1 h, respectively. Moreover, AKS was compatible with a variety of fluorescence proteins and dyes. Most importantly, AKS enhanced the fluorescence of YFP, in contrast to the major...