Femtosecond near-infrared laser pulses as a versatile non-invasive tool for intra-tissue nanoprocessing in plants without compromising viability
Femtosecond near-infrared laser pulses as a versatile non-invasive tool for intra-tissue nanoprocessing in plants without compromising viability
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DOI：
10.1046/j.1365-313x.2002.01346.x
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发表时间：
2002-08-01
期刊：
影响因子：
7.2
通讯作者：
König, K
中科院分区：
文献类型：
--
作者：
Tirlapur, UK;König, K
在本报告中，我们描述了一种高度可重复的基于飞秒近红外（NIR）激光的纳米加工技术，该技术可用于植物细胞壁的非侵入性组织内纳米解剖，以及选择性破坏单个质体或其部分，而不影响细胞的活力。超精密组织内纳米处理是通过将近红外（λ = 740 和 800 nm）飞秒钛蓝宝石激光器的辐射衍射极限聚焦到亚飞升体积并随后形成高度局部化的瞬时等离子体来产生高光强度（10(12) W cm(-2) ）来实现的。纳米手术后，对相应细胞目标区域的电子显微镜分析显示，细胞壁上有干净的非交错损伤，切口宽度小于 400 nm。据我们所知，这是在植物组织中非侵入性进行的最小切割。进一步的证据，包括叶绿素荧光的双光子成像，表明使用强烈的超快近红外脉冲可以完全敲除单个目标叶绿体或其一部分，而不会对相邻质体产生任何可见的有害影响。通过从细胞中排除碘化丙啶以及细胞质流的存在，可以确定纳米处理后细胞的活力。这项技术进步“q1”的潜在应用包括发育生物学应用，特别是解决个体发生事件和细胞与细胞相互作用的时空控制的研究，以及引力生物学应用。
In this report, we describe a highly reproducible femtosecond near-infrared (NIR) laser-based nanoprocessing technique that can be used both for non-invasive intra-tissue nanodissection of plant cell walls as well as selective destruction of a single plastid or part thereof without compromising the viability of the cells. The ultra-precise intra-tissue nanoprocessing is achieved by the generation of high light intensity (10(12) W cm(-2) ) by diffraction-limited focusing of the radiation of an NIR (lambda = 740 and 800 nm) femtosecond titanium-sapphire laser to a sub-femtolitre volume and subsequent highly localized instantaneous plasma formation. Following nanosurgery, electron microscopical analysis of the corresponding cellular target areas revealed clean non-staggering lesions across the cell wall with a cut width measuring less than 400 nm. To our knowledge, this is the smallest cut made non-invasively within a plant tissue. Further evidence, including two-photon imaging of chlorophyll fluorescence, revealed that a single target chloroplast or part thereof can be completely knocked out using intense ultra-fast NIR pulses without any visible deleterious effect on the adjacent plastids. The vitality of the cells after nanoprocessing has been ascertained by exclusion of propidium iodide from the cells as well as by the presence of cytoplasmic streaming. The potential applications of this technical advance "q1" include developmental biology applications, particularly studies addressing spatio-temporal control of ontogenetic events and cell-cell interactions, and gravitational biology applications.