Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll
Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll
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DOI：
10.1111/j.1469-8137.2010.03244.x
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发表时间：
2010-01-01
期刊：
影响因子：
9.4
通讯作者：
Love, John
中科院分区：
文献类型：
--
作者：
Littlejohn, George R.;Gouveia, Joao D.;Love, John
叶肉中的空气空间产生有害的光学效应，影响共聚焦显微镜。将叶片置于无毒、无荧光的全氟碳、全氟十烷（PFD）中，利用共聚焦显微镜和叶绿素荧光技术评估其光学增强和生理效果。在PFD中安装拟南芥叶片显著改善了叶片的光学质量，从而使高分辨率激光扫描共聚焦成像比用水更深入叶肉两倍。在PFD中孵育对挂载标本的生理影响小于水。我们得出结论，应用PFD作为安装介质，大大提高了活叶肉细胞和维管束细胞的共聚焦图像分辨率，而生理影响最小。
P>Air spaces in the leaf mesophyll generate deleterious optical effects that compromise confocal microscopy.Leaves were mounted in the nontoxic, nonfluorescent perfluorocarbon, perfluorodecalin (PFD), and optical enhancement and physiological effect were assessed using confocal microscopy and chlorophyll fluorescence.Mounting leaves of Arabidopsis thaliana in PFD significantly improved the optical qualities of the leaf, thereby enabling high-resolution laser scanning confocal imaging over twofold deeper into the mesophyll, compared with using water. Incubation in PFD had less physiological impact on the mounted specimen than water.We conclude that the application of PFD as a mounting medium substantially increases confocal image resolution of living mesophyll and vascular bundle cells, with minimal physiological impact.