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Negatively-Chirped Laser Enables Nonlinear Excitation and Nanoprocessing with sub-20-fs Pulses

机译：负hir激光可实现亚20 fs脉冲的非线性激发和纳米处理

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It has long been considered that the advantages emerging from employing chirp pre-compensation in nonlinear microscopy were overweighed by the complexity of prism- or grating-based compressors. These concerns were refuted with the advent of dispersive-mirrors-based compressors that are compact, user-friendly and sufficiently accurate to support sub-20-fs pulse delivery. Recent advances in the design of dispersive multilayer mirrors resulted in improved bandwidth (covering now as much as half of the gain bandwidth of Ti: Sapphire) and increased dispersion per bounce (one reflection off a state-of-the-art dispersive mirror pre-compensates the dispersion corresponding to > 10mm of glass). The compressor built with these mirrors is sufficiently compact to be integrated in the housing of a sub-12-fs Ti: Sapphire oscillator. A complete scanning nonlinear microscope (FemtOgene, JenLab GmbH) equipped with highly-dispersive, large-NA objectives (Zeiss EC Plan-Neofluoar 40x/1.3, Plan-Neofluar 63x/1,25 Oil) was directly seeded with this negatively chirped laser. The pulse duration was measured at the focus of the objectives by inserting a scanning autocorrelator in the beam path between the laser and the microscope and recording the second order interferometric autocorrelation traces with the detector integrated in the microscope. Pulse durations < 20fs were measured with both objectives. The system has been applied for two-photon imaging, transfection and optical manipulation of stem cells. Here we report on the successful transfection of human stem cells by transient optoporation of the cell membrane with a low mean power of < 7 mW and a short us beam dwell time. Optically transfected cells were able to reproduce. The daughter cell expressed also green fluorescent proteins (GFP) indicating the successful modification of the cellular DNA.
机译：长期以来一直认为，在非线性显微镜中采用线性调频预补偿所产生的优势被基于棱镜或光栅的压缩机的复杂性所抵消。基于分散镜的压缩机的问世驳斥了这些担忧，该压缩机结构紧凑，用户友好且足够精确以支持20 fs以下的脉冲传输。色散多层镜设计的最新进展提高了带宽（现在覆盖了钛：蓝宝石的增益带宽的一半），并提高了每反弹的色散（从最先进的色散镜预反射得到的反射）补偿大于10mm玻璃的色散）。装有这些反射镜的压缩机非常紧凑，可以集成在12-fs以下的Ti：蓝宝石振荡器的外壳中。带有负-激光直接播种了配备了高度分散的大型NA物镜（Zeiss EC Plan-Neofluoar 40x / 1.3，Plan-Neofluar 63x / 1,25油）的完整扫描非线性显微镜（FemtOgene，JenLab GmbH）。通过在激光和显微镜之间的光路中插入扫描自相关器并使用集成在显微镜中的检测器记录二阶干涉自相关迹线，来测量物镜焦点处的脉冲持续时间。两个目标均测量了<20fs的脉冲持续时间。该系统已应用于干细胞的双光子成像，转染和光学操作。在这里，我们报道了通过干膜细胞膜的瞬时平衡成功实现了人类干细胞的转染，细胞膜的平均功率<7 mW，信号束停留时间短。光学转染的细胞能够繁殖。子细胞还表达绿色荧光蛋白（GFP），表明细胞DNA的成功修饰。

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