Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel.
Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel.
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DOI：
10.3791/55044
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发表时间：
2017-01-18
期刊：
影响因子：
--
通讯作者：
Friml J
中科院分区：
文献类型：
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作者：
von Wangenheim D;Hauschild R;Friml J
理解植物发育的关键问题之一是单个细胞在组织的大背景下如何表现。因此，它需要在长时间内以高空间和时间分辨率观察整个器官，这可能会引起光毒性效应。该方案展示了一种用于光片显微镜的植物样品制备方法，其特点是将植物垂直安装在凝胶表面上。植物以这样一种方式安装，根部淹没在液体介质中，而叶子留在空气中。为了保证植物的光合作用，一个定制的照明系统照亮了叶子。为了使根保持在黑暗中，水面上覆盖着黑色塑料箔片。这种方法允许在标准化条件下对植物器官发育进行长期成像。
One of the key questions in understanding plant development is how single cells behave in a larger context of the tissue. Therefore, it requires the observation of the whole organ with a high spatial- as well as temporal resolution over prolonged periods of time, which may cause photo-toxic effects. This protocol shows a plant sample preparation method for light-sheet microscopy, which is characterized by mounting the plant vertically on the surface of a gel. The plant is mounted in such a way that the roots are submerged in a liquid medium while the leaves remain in the air. In order to ensure photosynthetic activity of the plant, a custom-made lighting system illuminates the leaves. To keep the roots in darkness the water surface is covered with sheets of black plastic foil. This method allows long-term imaging of plant organ development in standardized conditions.