Visualization of plastids and lipophilic components in living colonies of a wild strain of the hydrocarbon-forming green alga Botryococcus by laser scanning confocal microscopy
Visualization of plastids and lipophilic components in living colonies of a wild strain of the hydrocarbon-forming green alga Botryococcus by laser scanning confocal microscopy
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DOI：
10.1023/a:1008055613827
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发表时间：
1998-01-01
影响因子：
3.3
通讯作者：
Cleary, AL
中科院分区：
文献类型：
--
作者：
Beakes, GW;Cleary, AL
来自澳大利亚伯利格里芬湖的产烃绿藻芽孢球菌的野生菌株的菌落用共聚焦激光扫描显微镜进行了检测。显微镜配有双波长氪氩激光器，可同时检测叶绿素自身荧光和亲脂性染料荧光。这种快速而简单的技术揭示了活细胞中自身荧光质体的精确结构构象及其在致密球形菌落内的三维空间排列。DIOC6(3)细胞被亲脂性碳氰染料染色，细胞的顶端包含强烈染色的颗粒阵列，以及一个更分散的内部池系统，被认为是内质网。阳离子亲脂性染料罗丹明123在最外层细胞质中显示出较细的网状系统，部分覆盖在质体上。两种染料都显示了细胞外基质的亲脂性，并使从菌落渗出的脂球的分泌可见。这表明，共聚焦激光扫描显微技术将成为一种理想的工具，用于筛选分离物形成和分泌碳氢化合物的潜力，这些过程在这种具有潜在商业价值的藻类中仍远未被清楚地了解。
Colonies of a wild strain from Lake Burley-Griffin, Australia, of the hydrocarbon-producing green alga Botryococcus were examined by confocal laser scanning microscopy. The microscope was fitted with a dual wavelength krypton-argon laser, which permitted simultaneous detection of chlorophyll autofluorescence and lipophilic dye fluorescence. This quick and simple technique revealed the precise structural conformation of the autofluorescing plastids in living cells and their 3-dimensional spatial arrangement within the dense globular colonies. Cells stained with the lipophilic carbocyanine dye, DIOC6(3) contain an apical array of intensely staining granules as well as a more diffuse internal cisternal system thought to be endoplasmic reticulum. The cationic lipophilic dye rhodamine123 revealed a finer reticulate system in the outermost cytoplasm partially overlaying the plastid. Both dyes revealed the lipophilic nature of the extracellular matrix and enabled the secretion of lipid globules exuded from the colonies to be visualized. It is suggested that confocal laser scanning microcopy would make an ideal tool to screen isolates for their potential to form and secrete hydrocarbon, processes which are still far from clearly understood in this potentially commercially important alga.