Whole-Brain Imaging with Single-Cell Resolution Using Chemical Cocktails and Computational Analysis

被引：945

作者：

Susaki, Etsuo A. ^{[1
,2
,3
,4
]}

Tainaka, Kazuki ^{[1
,3
,4
]}

Perrin, Dimitri ^{[2
]}

Kishino, Fumiaki ^{[5
]}

Tawara, Takehiro ^{[6
]}

Watanabe, Tomonobu M. ^{[7
]}

Yokoyama, Chihiro ^{[8
]}

Onoe, Hirotaka ^{[8
]}

Eguchi, Megumi ^{[9
]}

Yamaguchi, Shun ^{[9
,10
]}

Abe, Takaya ^{[11
]}

Kiyonari, Hiroshi ^{[11
]}

Shimizu, Yoshihiro ^{[12
]}

Miyawaki, Atsushi ^{[13
]}

Yokota, Hideo ^{[6
]}

Ueda, Hiroki R. ^{[1
,2
,3
,4
]}

机构：

[1] RIKEN Quantitat Biol Ctr, Lab Synthet Biol, Chuo Ku, Kobe, Hyogo 6500047, Japan

[2] RIKEN Ctr Dev Biol, Lab Syst Biol, Chuo Ku, Kobe, Hyogo 6500047, Japan

[3] Univ Tokyo, Grad Sch Med, Dept Syst Pharmacol, Bunkyo Ku, Tokyo 1130033, Japan

[4] Japan Sci & Technol Agcy, CREST, Kawaguchi, Saitama 3320012, Japan

[5] Univ Tokyo, Fac Med, Bunkyo Ku, Tokyo 1130033, Japan

[6] RIKEN Ctr Adv Photon, Image Proc Res Team, Wako, Saitama 3510198, Japan

[7] Osaka Univ, OLABB, RIKEN Quantitat Biol Ctr, Lab Comprehens Bioimaging, Suita, Osaka 5650874, Japan

[8] RIKEN Ctr Life Sci Technol, Biofunct Imaging Team, Chuo Ku, Kobe, Hyogo 6500047, Japan

[9] Gifu Univ, Grad Sch Med, Div Morphol Neurosci, Gifu 5011194, Japan

[10] Japan Sci & Technol Agcy, PRESTO, Kawaguchi, Saitama 3320012, Japan

[11] RIKEN Ctr Dev Biol, Lab Anim Resources & Genet Engn, Chuo Ku, Kobe, Hyogo 6500047, Japan

[12] RIKEN Quantitat Biol Ctr, Lab Cell Free Prot Synth, Chuo Ku, Kobe, Hyogo 6500047, Japan

[13] RIKEN Brain Sci Inst, Lab Cell Funct Dynam, Wako, Saitama 3510198, Japan

来源：

CELL | 2014年 / 157卷 / 03期

基金：

日本学术振兴会; 日本科学技术振兴机构;

关键词：

LIGHT-SHEET MICROSCOPY; MOUSE-BRAIN; IN-VIVO; ULTRAMICROSCOPY; RECONSTRUCTION; CONNECTIONS; PROJECTIONS; EXPRESSION; MICE; VISUALIZATION;

D O I：

10.1016/j.cell.2014.03.042

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Systems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution.

引用

页码：726 / 739

页数：14

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