Visualizing Plant Cells in A Brand New Way
Visualizing Plant Cells in A Brand New Way
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以全新方式可视化植物细胞
DOI：
10.1016/j.molp.2016.02.006
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发表时间：
--
期刊：
影响因子：
27.5
通讯作者：
Hao Wang
中科院分区：
文献类型：
--
作者：
Hao Wang
荧光标记的融合蛋白的体内成像对于细胞生物学家研究活标本中的蛋白质定位、动力学和功能是必不可少的。不同的视觉技术，从经典的激光共聚焦扫描显微镜到新开发的超分辨率显微镜，都极大地帮助生物学家做出了新的发现。在植物中，成像技术的应用往往受到植物挑战性特性的限制，这些特性包括：(I)由于多层细胞堆叠和厚厚的植物细胞壁而导致的样品不透明；(Ii)来自叶绿素、木质素、叶黄素和外部蜡质表面的强烈自发荧光干扰。近年来，晶格光学显微镜的突破性发展显示了在分子水平上对活细胞或组织进行四维成像的独特能力，而背景和干扰可以忽略不计。通过利用超薄和无衍射激发贝塞尔光束的优势，它显示出超高的时空分辨率，可以跟踪纳米尺度的亚细胞过程，良好的样品穿透能力，以及超低的光毒性和光漂白，这是特别理想的长期实时成像。因此，晶格薄片显微镜不是更新成像设备的问题，而是为活体标本的4D视觉技术开辟了一个全新的未来，克服了植物材料的局限性。因此，这预示着植物细胞成像的一个很有前途的新未来。
In vivo imaging of fluorescence-tagged fusion proteins is essential to study protein localization, dynamics, and functions in living specimens for cell biologists. Different visual technologies, ranging from classical confocal laser scanning microscopy to newly developed super-resolution microscopy, have tremendously assisted biologists in making new discoveries. In plants, application of imaging technologies is often limited due to plant challenging properties which include:(i) opacity of specimens due to multiple stack layers of cells and a thick plant cell wall; and (ii) strong autofluorescence disturbance from chlorophyll, lignin, xanthophyll, and an outer waxy surface. Recent groundbreaking development of lattice light-sheet microscopy has demonstrated unique abilities to image live cells or tissues in four-dimensions (4D) at molecular level with negligible background and interference. By taking advantages of ultrathin and non-diffracting excitation Bessel beams, it shows extraordinary high spatiotemporal resolution to follow nanoscale subcellular processes, good sample-penetration ability, and ultralow phototoxicity and photobleaching, which are particularly ideal for longterm live imaging. Therefore, lattice light-sheet microscopy is not a matter of updating the imaging facilities but opens up a brand new future for 4D visual technology of live specimens and overcomes the limitations of plant material. It thus suggests a promising new future for plant cell imaging.