Fast free-of-acrylamide clearing tissue (FACT)-an optimized new protocol for rapid, high-resolution imaging of three-dimensional brain tissue.
Fast free-of-acrylamide clearing tissue (FACT)-an optimized new protocol for rapid, high-resolution imaging of three-dimensional brain tissue.
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快速无丙烯酰胺透明组织 (FACT)——一种优化的新方案，用于三维脑组织的快速、高分辨率成像
DOI：
10.1038/s41598-017-10204-5
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发表时间：
2017-08-29
影响因子：
4.6
通讯作者：
Feng Y
中科院分区：
文献类型：
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作者：
Xu N;Tamadon A;Liu Y;Ma T;Leak RK;Chen J;Gao Y;Feng Y
快速无丙烯酰胺清除组织(FACT)是一种新的基于十二烷基硫酸钠(十二烷基硫酸钠)的清除方案，用于含有转基因或免疫标记荧光蛋白的脑组织的化学清除和成像。在本研究中，我们开发了这一新方法，并优化了工作流程的多个维度，包括减少清除时间、提高荧光信号的效率而不需要电泳或复杂仪器、保留细胞结构细节、优化共聚焦显微镜以及加速数据收集。我们使用在小胶质细胞中表达荧光色素的转基因小鼠大脑，系统地比较了七种清除方案与FACT方案。在1毫米厚的成年小鼠脑片中检测转基因标记标记只需要6天，在相同大小的组织中检测抗体标记标记需要14天。通过减少清除时间、调节十二烷基硫酸钠溶液的pH值、采用合适的温度进行组织清除，实现了荧光信号的保存，这些都是该方法的优越性所在。我们的结论是，FACT协议可以成功地应用于小鼠脑组织的荧光成像，并将有助于在三维器官系统的背景下进行大型细胞及其网络组装的结构分析和连接。
Fast Free-of-Acrylamide Clearing Tissue (FACT) is a new sodium dodecyl sulfate (SDS)-based clearing protocol for the chemical clearing and imaging of brain tissue containing transgenic or immunolabeled fluorescent proteins. In the present study, we have developed this new method and optimized multiple dimensions of the workflow, including reduced clearing time, improved efficiency of fluorescent signals without the need for electrophoretic or complex instrumentations, preservation of cytoarchitectural details, optimized confocal microscopy, and accelerated data collection. We systematically compared seven clearing protocols with the FACT protocol, using transgenic mouse brains with fluorochrome expression in microglia. Only six days were required for detecting transgene-labeled markers in a 1-mm thick brain slice from adult mice, and 14 days were required for detecting antibody-labeled markers in the same-sized tissue. Preservation of fluorescent signal was achieved by decreasing clearing time, adjusting the pH of the SDS solution, and using the appropriate temperature for tissue clearing, all of which contributed to the superiority of our method. We conclude that the FACT protocol can be successfully applied to the fluorescent imaging of mouse brain tissue, and will facilitate structural analyses and connectomics of large assemblies of cells and their networks in the context of three-dimensional organ systems.
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影响因子：
48
作者：
Chung, Kwanghun;Deisseroth, Karl
通讯作者：
Deisseroth, Karl
DOI：
10.1177/1073858413504465
发表时间：
2014-10
期刊：
The Neuroscientist : a review journal bringing neurobiology, neurology and psychiatry
影响因子：
--
作者：
Fields RD;Araque A;Johansen-Berg H;Lim SS;Lynch G;Nave KA;Nedergaard M;Perez R;Sejnowski T;Wake H
通讯作者：
Wake H
简单，可扩展的蛋白质组学成像，用于完整系统的高维分析。
影响因子：
64.5
作者：
Murray E;Cho JH;Goodwin D;Ku T;Swaney J;Kim SY;Choi H;Park YG;Park JY;Hubbert A;McCue M;Vassallo S;Bakh N;Frosch MP;Wedeen VJ;Seung HS;Chung K
通讯作者：
Chung K
影响因子：
64.8
作者：
通讯作者：
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影响因子：
14.8
作者：
通讯作者：
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