Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues.
Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues.
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DOI：
10.1038/nbt.3641
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发表时间：
2016-09
影响因子：
46.9
通讯作者：
Chung, Kwanghun
中科院分区：
文献类型：
--
作者：
Ku, Taeyun;Swaney, Justin;Park, Jeong-Yoon;Albanese, Alexandre;Murray, Evan;Cho, Jae Hun;Park, Young-Gyun;Mangena, Vamsi;Chen, Jiapei;Chung, Kwanghun
多细胞生物体的生物学在多个尺寸尺度上协调，从亚纳米级的分子到宏观尺度，细胞群体的组织范围内的相互连接。在这里，我们介绍了一种方法的多尺度组织的完整组织的超分辨率成像。该方法被称为蛋白质组放大分析（MAP），将整个器官线性扩展四倍，同时保留其整体结构和三维蛋白质组组织。MAP基于以下观察结果：在水凝胶-组织杂交期间防止内源性蛋白质内和内源性蛋白质之间的交联允许在蛋白质变性和解离时自然扩增。扩张的组织保留了其蛋白质含量，其精细的亚细胞细节和其器官规模的细胞间连接。现成的抗体可用于组织放大蛋白质组的多轮免疫标记和成像，我们的实验表明成功率为82%（测试的100/122抗体）。我们发现，标本的大小可以可逆地调制图像区域间的连接和精细的突触结构在小鼠大脑。
The biology of multicellular organisms is coordinated across multiple size scales, from the sub-nanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs four-fold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details and its organ-scale intercellular connectivity. Off-the-shelf antibodies can be used for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, with our experiments demonstrating a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
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光学成像。膨胀显微镜。
DOI：
10.1126/science.1260088
发表时间：
2015-01-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen F;Tillberg PW;Boyden ES
通讯作者：
Boyden ES
影响因子：
48
作者：
Craddock, R. Cameron;Jbabdi, Saad;Yan, Chao-Gan;Vogelstein, Joshua T.;Castellanos, F. Xavier;Di Martino, Adriana;Kelly, Clare;Heberlein, Keith;Colcombe, Stan;Milham, Michael P.
通讯作者：
Milham, Michael P.
影响因子：
7.7
作者：
Albanese A;Chung K
通讯作者：
Chung K
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
48
作者：
Chung, Kwanghun;Deisseroth, Karl
通讯作者：
Deisseroth, Karl