Department of Systems Pharmacology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Laboratory for Synthetic Biology, RIKEN Quantitative Biology Center, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan; CREST, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
Department of Systems Pharmacology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Cell. 2014 Nov 6;159(4):911-24. doi: 10.1016/j.cell.2014.10.034.
The development of whole-body imaging at single-cell resolution enables system-level approaches to studying cellular circuits in organisms. Previous clearing methods focused on homogenizing mismatched refractive indices of individual tissues, enabling reductions in opacity but falling short of achieving transparency. Here, we show that an aminoalcohol decolorizes blood by efficiently eluting the heme chromophore from hemoglobin. Direct transcardial perfusion of an aminoalcohol-containing cocktail that we previously termed CUBIC coupled with a 10 day to 2 week clearing protocol decolorized and rendered nearly transparent almost all organs of adult mice as well as the entire body of infant and adult mice. This CUBIC-perfusion protocol enables rapid whole-body and whole-organ imaging at single-cell resolution by using light-sheet fluorescent microscopy. The CUBIC protocol is also applicable to 3D pathology, anatomy, and immunohistochemistry of various organs. These results suggest that whole-body imaging of colorless tissues at high resolution will contribute to organism-level systems biology.
单细胞分辨率的全身成像技术使我们能够采用系统生物学的方法来研究生物体中的细胞回路。先前的透明化方法主要侧重于均匀化单个组织之间不匹配的折射率,从而降低了不透明度,但仍未达到完全透明的效果。在这里,我们发现一种氨基醇通过有效地从血红蛋白中洗脱血红素发色团来使血液褪色。我们之前将含有氨基醇的鸡尾酒称为 CUBIC,并通过直接心脏灌注该鸡尾酒,然后再结合 10 天至 2 周的透明化方案,可使成年小鼠的几乎所有器官以及新生和成年小鼠的整个身体褪色并变得几乎完全透明。该 CUBIC 灌注方案通过使用光片荧光显微镜能够快速实现单细胞分辨率的全身和全器官成像。CUBIC 方案也适用于各种器官的 3D 病理学、解剖学和免疫组织化学。这些结果表明,高分辨率的无色组织全身成像将有助于机体系统生物学的研究。