Application of confocal laser scanning microscopy in plant biology
Application of confocal laser scanning microscopy in plant biology
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共聚焦激光扫描显微镜在植物生物学中的应用
DOI：
10.1002/bbpc.19890930332
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发表时间：
1989
影响因子：
3
通讯作者：
H. Engelhardt
中科院分区：
文献类型：
--
作者：
W. Knebel;H. Quader;R. Wijnaendts;H. Engelhardt
荧光共聚焦激光扫描显微镜（CLSM）越来越被认为是研究动植物细胞三维结构的一种合适的技术。成像过程包括收集物体中一定体积的发出的荧光。然而，需要一个强大的处理系统来存储单个数据并重建对象的图像。物体的三维图像可以通过扩展聚焦、立体图像或阴影模型来实现。-这些程序用于详细说明固定或活植物细胞中结构成分和细胞器的空间排列。用间接免疫荧光技术观察了洋葱根细胞和苔藓原胞茎尖细胞分裂过程中的微管排列。此外，内质网通过荧光染料3,3'-二己氧碳氰碘（DiOC）的活体染色可见。例如在正面洋葱鳞茎鳞片表皮细胞和根毛细胞。此外，还展示了几个例子，证明了CLSM在单个细胞器的放大，图像和运动测量方面提供的可能性。一些测量是用一个紧凑的反向共聚焦系统进行的，该系统还包括一个经典的荧光显微镜。荧光共聚焦激光扫描显微镜由于具有分辨聚焦深度以外的荧光灯的特性，在研究植物细胞成分的三维排列方面具有很大的优势。
Fluorescence confocal laser scanning microscopy (CLSM) is increasingly acknowledged as a suitable technique to study the 3-dimensional structure of animal and plant cells. The imaging process involves the collection of the emitted fluorescent light of a defined volume in the object. A powerful processing system is, however, required to store the single data and reconstruct the image of the object. A 3-D-image of an object can be achieved either by the extended focus, the stereo image, or the shadowing mode1). —These procedures were applied to elaborate on the spatial arrangement of structural components and organells in fixed or living plant cells. Microtubule arrays in dividing onion root cells and in caulonema tip cells of moss protonemata are visualized by the indirect immunofluorescence technique. In addition, the endoplasmic reticulum is visualized through vital staining with the fluorochrome 3,3'-dihexyloxacarbocyanine-iodide (DiOC). For example in adaxial onion bulb scale epidermis cells and root hair cells. Also, several examples are shown which demonstrate the possibilities the CLSM offers with respect to magnification, image, and motility measurements of single organelles. Some of the measurements have been performed with a compact inverted confocal system which also includes a classical fluorescence microscope. Fluorescence confocal laser scanning microscopy turned out to be of great advantage for studying the 3-D-arrangement of plant cell constituents due to the property of the focal scanner which discriminates fluorescent light outside of the depth of focus.