Broad applicability of a streamlined ethyl cinnamate-based clearing procedure
Broad applicability of a streamlined ethyl cinnamate-based clearing procedure
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DOI：
10.1242/dev.166884
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发表时间：
2019-02-01
期刊：
影响因子：
4.6
通讯作者：
Tanaka, Elly M.
中科院分区：
文献类型：
--
作者：
Masselink, Wouter;Reumann, Daniel;Tanaka, Elly M.
混浊和不透明是组织的固有特性，其限制了通过大组织获取显微图像的能力。创建均匀的折射率，称为组织清除，克服了大多数这些问题。这些方法使研究人员能够以前所未有的深度和分辨率对大型复杂的3D结构进行成像。然而，由于成本、时间、现有方法的复杂性和对荧光信号的潜在负面影响的组合，组织清除在有限程度上被采用。在这里，我们描述了2 Eci（第二代基于肉桂酸乙酯的清除），它可用于在短短1-5天内清除几个物种的各种组织，包括人类类器官，黑腹果蝇，斑马鱼，蝾螈和非洲爪蟾，同时保留广泛的荧光蛋白，包括GFP，mCherry，Brainbow和Alexa共轭荧光团。肉桂酸乙酯是无毒的，可以很容易地用于多用户显微镜设施。这种方法由于其易于使用，肉桂酸乙酯的无毒性质和广泛的适用性，为更广泛的研究人员开放了组织清除。
Turbidity and opaqueness are inherent properties of tissues that limit the capacity to acquire microscopic images through large tissues. Creating a uniform refractive index, known as tissue clearing, overcomes most of these issues. These methods have enabled researchers to image large and complex 3D structures with unprecedented depth and resolution. However, tissue clearing has been adopted to a limited extent due to a combination of cost, time, complexity of existing methods and potential negative impact on fluorescence signal. Here, we describe 2Eci (2nd generation ethyl cinnamate-based clearing), which can be used to clear a wide range of tissues in several species, including human organoids, Drosophila melanogaster, zebrafish, axolotl and Xenopus laevis, in as little as 1-5 days, while preserving a broad range of fluorescent proteins, including GFP, mCherry, Brainbow and Alexa-conjugated fluorophores. Ethyl cinnamate is non-toxic and can easily be used in multi-user microscope facilities. This method opens up tissue clearing to a much broader group of researchers due to its ease of use, the non-toxic nature of ethyl cinnamate and broad applicability.