ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging.
ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging.
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DOI：
10.1242/dev.127613
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发表时间：
2015-12-01
期刊：
影响因子：
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通讯作者：
Higashiyama T
中科院分区：
文献类型：
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作者：
Kurihara D;Mizuta Y;Sato Y;Higashiyama T
用于可视化和分析精确形态和基因表达模式的成像技术对于理解所有生物体发育过程中的生物过程至关重要。在化学筛选的帮助下，我们开发了一种使用化学溶液的清洁方法，称为ClearSee，用于植物组织中形态和基因表达的深度成像。ClearSee可迅速减少叶绿素的自发荧光，同时保持荧光蛋白的稳定性。通过调节折射率失配，可以在ClearSee处理的样品中通过共焦和双光子激发显微镜进行全器官和全植物成像。此外，ClearSee还适用于荧光蛋白的荧光成像，以允许对多个基因表达进行结构分析。鉴于ClearSee与化学染料染色兼容，该技术可用于与遗传标记结合的深度成像以及不适合转基因方法的植物物种。该方法适用于完整形态的整体成像，有助于加速植物生物学研究中新现象的发现。总结：光学透明试剂ClearSee改善了荧光蛋白和染料的荧光成像，并允许对多种植物组织中的基因表达模式进行结构分析。
Imaging techniques for visualizing and analyzing precise morphology and gene expression patterns are essential for understanding biological processes during development in all organisms. With the aid of chemical screening, we developed a clearing method using chemical solutions, termed ClearSee, for deep imaging of morphology and gene expression in plant tissues. ClearSee rapidly diminishes chlorophyll autofluorescence while maintaining fluorescent protein stability. By adjusting the refractive index mismatch, whole-organ and whole-plant imaging can be performed by both confocal and two-photon excitation microscopy in ClearSee-treated samples. Moreover, ClearSee is applicable to multicolor imaging of fluorescent proteins to allow structural analysis of multiple gene expression. Given that ClearSee is compatible with staining by chemical dyes, the technique is useful for deep imaging in conjunction with genetic markers and for plant species not amenable to transgenic approaches. This method is useful for whole imaging for intact morphology and will help to accelerate the discovery of new phenomena in plant biological research. Summary: The optical clearing reagent ClearSee improves the multicolor imaging of fluorescent proteins and dyes and allows the structural analysis of gene expression patterns in multiple plant tissues.