Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing

被引：790

作者：

Yang, Bin ^{[1
]}

Treweek, Jennifer B. ^{[1
]}

Kulkarni, Rajan P. ^{[1
,2
]}

Deverman, Benjamin E. ^{[1
]}

Chen, Chun-Kan ^{[1
]}

Lubeck, Eric ^{[1
]}

Shah, Sheel ^{[1
]}

Cai, Long ^{[3
]}

Gradinaru, Viviana ^{[1
]}

机构：

[1] CALTECH, Div Biol & Biol Engn, Pasadena, CA 91125 USA

[2] Univ Calif Los Angeles, David Geffen Sch Med, Dept Med, Div Dermatol, Los Angeles, CA 90095 USA

[3] CALTECH, Div Chem & Chem Engn, Pasadena, CA 91125 USA

来源：

CELL | 2014年 / 158卷 / 04期

关键词：

MESSENGER-RNA DETECTION; NEURONS; VISUALIZATION; VECTORS; CLARITY; ORGANS;

D O I：

10.1016/j.cell.2014.07.017

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. We describe PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We show that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution. These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies.

引用

页码：945 / 958

页数：14

共 31 条

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