Femtosecond laser nanosurgery of sub-cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool
Femtosecond laser nanosurgery of sub-cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool
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DOI：
10.1002/jbio.201100055
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发表时间：
2012-02-01
影响因子：
2.8
通讯作者：
Filippidis, George
中科院分区：
文献类型：
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作者：
Tserevelakis, George J.;Psycharakis, Stylianos;Filippidis, George
飞秒激光辅助显微生物标本的纳米手术是一种相对较新的技术，它允许选择性地破坏亚细胞结构，而不会对周围区域造成任何不希望的损伤。必须对目标结构进行染色，以便在纳米外科手术中清晰可见。然而，最终的纳米手术结果的验证是困难的，因为目标结构可以简单地光漂白，而不是选择性地破坏。这一事实构成了该技术的主要缺点。在我们的研究中，我们采用了一种多模式系统，该系统将非线性成像模式与纳米手术能力相结合，用于选择性破坏HeLa癌细胞中的亚细胞结构。三次谐波发生（THG）成像模式被用作一种工具，用于识别的结构，进行纳米手术实验。不需要对生物样品进行染色，因为THG是物质的固有性质。此外，通过双光子激发荧光（TPEF）测量验证了纳米手术处理后的细胞活力。(C)2012 WILEY-VCH Verlag GmbH & Co. KGaA，魏因海姆）
Femtosecond laser assisted nanosurgery of microscopic biological specimens is a relatively new technique which allows the selective disruption of sub-cellular structures without causing any undesirable damage to the surrounding regions. The targeted structures have to be stained in order to be clearly visualized for the nanosurgery procedure. However, the validation of the final nanosurgery result is difficult, since the targeted structure could be simply photobleached rather than selectively destroyed. This fact comprises a main drawback of this technique. In our study we employed a multimodal system which integrates non-linear imaging modalities with nanosurgery capabilities, for the selective disruption of sub-cellular structures in HeLa cancer cells. Third Harmonic Generation (THG) imaging modality was used as a tool for the identification of structures that were subjected to nanosurgery experiments. No staining of the biological samples was required, since THG is an intrinsic property of matter. Furthermore, cells' viability after nanosurgery processing was verified via Two Photon Excitation Fluorescence (TPEF) measurements. (C) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)