Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine.
Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine.
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DOI：
10.1186/s12861-014-0048-3
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发表时间：
2014-12-21
影响因子：
--
通讯作者：
Kim S
中科院分区：
文献类型：
--
作者：
Lee H;Park JH;Seo I;Park SH;Kim S
在细胞，电路和器官范围内绘制组织结构对于理解生理和生物功能是重要的。最近开发了一种称为CLARITY的生物电化学技术，用于透明完整组织内的三维解剖和表型定位。该方法为理解神经系统和器官回路的结构-功能关系提供了重大进展。因此，在本研究中，我们旨在改进原始的CLARITY程序，并为各种完整器官制定特定的CLARITY方案。我们确定了减少气泡形成、变色和组织表面黑色颗粒沉积的最佳条件，从而可以产生更清晰的器官图像。我们还确定了每种类型器官合适的清除液更换周期，并令人信服地证明了250-280 mA是组织清除的理想电流范围。然后，我们获得了各种类型的清除器官，包括脑、胰腺、肝脏、肺、肾和肠。此外，我们还测定了海马区轴突纤维、小脑浦肯野层、胰腺血管和细胞核的图像。CLARITY是一项创新的生化技术，用于各种类型组织的结构和分子分析。我们开发了用于清除脑、胰腺、肺、肠、肝和肾的改进CLARITY方法，并确定了清除每种特定组织类型的适当实验条件。这些优化的方法将有助于CLARITY在不同类型器官中的应用。本文的在线版本（doi:10.1186/s12861-014-0048-3）包含补充材料，授权用户可以使用。
Mapping of tissue structure at the cellular, circuit, and organ-wide scale is important for understanding physiological and biological functions. A bio-electrochemical technique known as CLARITY used for three-dimensional anatomical and phenotypical mapping within transparent intact tissues has been recently developed. This method provided a major advance in understanding the structure-function relationships in circuits of the nervous system and organs by using whole-body clearing. Thus, in the present study, we aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs. We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images. We also determined the appropriate replacement cycles of clearing solution for each type of organ, and convincingly demonstrated that 250–280 mA is the ideal range of electrical current for tissue clearing. We then acquired each type of cleared organs including brain, pancreas, liver, lung, kidney, and intestine. Additionally, we determined the images of axon fibers of hippocampal region, the Purkinje layer of cerebellum, and vessels and cellular nuclei of pancreas. CLARITY is an innovative biochemical technology for the structural and molecular analysis of various types of tissue. We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type. These optimized methods will be useful for the application of CLARITY to various types of organs. The online version of this article (doi:10.1186/s12861-014-0048-3) contains supplementary material, which is available to authorized users.
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影响因子：
64.8
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14.8
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影响因子：
29
作者：
Rodriguez-Diaz R;Abdulreda MH;Formoso AL;Gans I;Ricordi C;Berggren PO;Caicedo A
通讯作者：
Caicedo A
影响因子：
3.2
作者：
SKALLI, O;PELTE, MF;ORCI, L
通讯作者：
ORCI, L
影响因子：
82.9
作者：
Speier, Stephan;Nyqvist, Daniel;Berggren, Per-Olof
通讯作者：
Berggren, Per-Olof