Realignment process of actin stress fibers in single living cells studied by focused femtosecond laser irradiation

Three-dimensional dissection of a single actin stress fiber in a living cell was performed based on multi-photon absorption of a focused femtosecond laser pulse. The realignment process of an actin stress fiber was investigated after its direct cutting by a single-shot femtosecond laser pulse irradiation by high-speed transmission and fluorescence imaging methods. It was confirmed that mechanical force led by the femtosecond laser cutting propagates to entire cell through the cytockelton in a 100 μs time scale. The cut actin stress fiber was realigned in the time scale of a few tens of minutes. The dynamic analysis of the realignment induced by single-shot femtosecond laser gives new information on cell activity.     

Measurement of UV absorption of single living cell for cell manipulation using NIR femtosecond laser

Optical UV absorption of single human living cells ranging from 200 nm to 360 nm was measured in situ for the study of cell manipulation using the near-infrared (NIR) femtosecond laser. Human breast living cells of MCF-10A, MCF-7, and MDA-MB-231 were used in this experiment. The selective photo-disruptions of single living cell and its sub-organelle (nucleus) were also demonstrated using the tightly focused 790 nm wavelength femtosecond laser with pulse duration of 110 fs. It was found that each living cell has its own absorption spectrum in UV wavelength ranges. It was also inferred that intrinsic absorption spectrum is attributed to the amount of DNA and protein of living cell. For the study of photo-disruption of single cell using the multi-photon absorption excited by the NIR femtosecond laser pulse, the origin UV absorption spectrum of targeted living cell is important and fundamental information to understand nonlinear interaction between NIR ultrashort, high-intensity laser light and transparent living cell.     

单个飞秒激光作用下熔融石英的微爆阈值研究

对飞秒激光脉冲在透明介质中产生微爆的物理机制进行了研究,比较了长短激光脉冲的光击穿机制,基于Fokker-Planck方程建立了飞秒激光微爆模型,给出了飞秒激光脉冲在透明介质中产生微爆的阈值解析表达式.针对飞秒激光脉冲在熔石英介质中的微爆阈值进行了计算,得到的结果与实验结果基本相符.     

THz generation via optical rectification with ultrashort laser pulse focused to a line

We report on efficient THz pulse generation via optical rectification with femtosecond laser pulses focused to a line by a cylindrical lens. This configuration provides phase-matched conditions in the superluminal regime. 35 pJ THz pulses have been generated with this technique in a stoichiometric LiNbO₃ crystal pumped by 2 μJ femtosecond laser pulses at room temperature. An unusual superquadratic rise of the THz pulse energy with the laser pulse energy has been observed at high laser energies. This extraordinary energy dependence of the THz generation efficiency is explained by self-focusing of the laser beam in the crystal. Z-scan measurements and comparison of the THz pulse spectra created with laser pulses having different energies confirm this interpretation.     

In situ observation of photo-bleaching in human single living cell excited by a NIR femtosecond laser

The photo-bleaching of single living cells excited by femtosecond laser irradiation was observed in situ to study the nonlinear interaction between ultrafast laser pulses and living human breast MDA-MB-231 cells. We conducted a systematic study of the energy dependence of plasma-mediated photo-disruption of fluorescently labeled subcellular structures in the nucleus of living cells using near-infrared (NIR) femtosecond laser pulses through a numerical aperture objective lens (0.75 NA). The behavior of photo-bleached living cells with fluorescently labeled nuclei was observed for 18 h after femtosecond laser irradiation under a fluorescence microscope. The photo-bleaching of single living cells without cell disruption occurred at between 470 and 630 nJ. To study the photo-disruption of subcellular organelles in single living cells using the nonlinear absorption excited by a NIR femtosecond laser pulse, the process of photo-bleaching without photo-disruption provides key information for clarifying the nonlinear interaction between NIR ultrashort, high-intensity laser light and transparent fluorescently labeled living cells.     

Dielectric microsphere mediated transfection using a femtosecond laser

We demonstrate the permeabilization of cell membranes by an enhanced optical field generated under polystyrene microspheres of 1000 nm diameter excited by a femtosecond laser pulse. Fluorescent molecules and short interfering RNA (siRNA) have been successfully delivered to many cells in the irradiated area by a single 80 fs laser pulse at 800 nm wavelength in the presence of antibody-conjugated polystyrene spheres. The ratios of the cells showing permeabilization were 38% and 21% for Fluorescein isothiocyanate-dextran and siRNA, respectively, at the laser fluence of 1.06 J/cm(2). The present method has advantages both in high throughput of many cell treatments and precise processing of minute areas on cell membranes.     

Nanoparticle injection to single animal cells using femtosecond laser-induced impulsive force

An impulsive force, which was generated by focusing tightly a femtosecond laser into a cell culture medium, was applied to inject nanoparticles into local areas of a single mouse fibroblast NIH3T3 cell. When the impulsive force was induced near the cell, the nanoparticles adhering on the cell membrane were introduced, which was directly confirmed by confocal fluorescence microscopy.     

Nonlinear optical effects of near-IR femtosecond laser radiation on the morphology and structure of a nerve cell in the field of an optical trap

Changes in the morphology and structure of a neuron cell in the process of manipulating it with an femtosecond laser tweezers operating at a wavelength of 800 nm are examined. The changes in the morphology and structure are caused by the nonlinear optical absorption and multiphoton excitation of cell biomacromolecules by femtosecond pulses of light. The cell nucleus is demonstrated to be destroyed by focused femtosecond laser radiation (FLR). Changes in the state of the cytoplasmic nucleoproteins and the hydrophobicity of the plasmatic membrane under the action of FLR focused inside the cell are observed. By the example of the simultaneous displacement of the neuron with a cw laser and cutting of a neuron with FLR, the operation of holographic optical manipulator and a scalpel based on the use of femtosecond and cw lasers is considered. The possibility of the simultaneous microsurgical operation with several optical foci of FLR is demonstrated.     

Redistribution of a material at femtosecond laser ablation of a thin silver film

Energy-dispersive X-ray microspectroscopy is used for the first time to quantitatively study the spatial displacement of the material of a 100-nm silver film irradiated by a single femtosecond laser pulse focused on a small spot in the diffraction limit. The silver mass distribution over radial cross sections is determined and matter balance is analyzed for the resulting radially symmetric submicron structures of a microcone with a nanospike with various heights and a through hole. Hydrodynamic processes and phase transitions inducing the melting of the film, motion of the melt, and its recrystallization within a focal spot are studied.     

Self-Assembled Quasi-Periodic Voids in Glass Induced by a Tightly Focused Femtosecond Laser

Multiple refocusing of a tightly focused femtosecond laser due to the dynamic transformation between self-focusing and self-defocusing is employed to provide a novel method to produce quasi-periodic voids in glass. It is found that the diameter or the interval of the periodic voids increases with the increasing pulse energy of the laser. The detailed course for producing periodic voids is discussed by analysing the damaged track induced by the tightly focused femtosecond laser pulses. It is suggested that this periodic structure has potential applications in fabrication of three-dimensional optical devices.     

飞秒激光烧蚀LiNbO3晶体的形貌特征与机理研究

采用了不同能量的单脉冲和多脉冲飞秒激光对LiNbO₃晶体进行烧蚀，并刻蚀了表面衍射型光栅.通过扫描电镜和原子力显微镜观察了烧蚀点的形貌特征，首次发现利用单束飞秒激光脉冲对LiNbO₃晶体烧蚀，可以得到超衍射极限的烧蚀点，当聚焦光斑直径约为2μm、能量为170nJ的单脉冲飞秒激光作用时，烧蚀点的直径约为400nm，100nJ，17个脉冲作用时烧蚀点的直径约为800nm.同时可以观察到在能量较低的多脉冲飞秒激光作用下， LiNbO₃晶体呈现出大约200nm周期性分布的波纹状结构.实验结果表明，选择合适参数的飞秒激光脉冲可以对LiNbO₃晶体进行超衍射极限加工，这对于利用飞秒激光制作LiNbO₃基质的微纳光电子器件有十分重要的意义.     

单次飞秒激光脉冲在玻璃内部产生多次微爆的研究

在入射能量不同的情况下，用近红外飞秒激光脉冲在重钡火石玻璃（ZBaF15）内部产生了色心和微爆，在熔融石英的内部产生了多次微爆现象.实验表明：入射能量较高时，色心的中间会产生微爆；在松聚焦条件下，一个飞秒激光脉冲在透明介质内会激发多次微爆.基于飞秒激光脉冲在透明介质内的自聚焦效应和自由电子等离子体的自散焦效应，理论分析了多次微爆产生的原因；也讨论了飞秒激光脉冲诱导玻璃折射率改变的机理.     

短脉冲激光加工双温方程的优化运算

为研究双温方程中电子热传导项和电子与晶格耦合项对激光辐照物体表面温度场求解的影响,对这两项施加了约束条件。由于飞秒和亚皮秒激光与物质相互作用时间短,电子与晶格来不及耦合,所以对耦合项施加时间约束;根据相分离条件(CPPS),对热传导项施加空间约束。利用有限元方法建立了激光烧蚀金属铜膜表面的有限元模型。通过分析双温方程中热传导项和耦合项对计算结果的影响,发现短脉冲激光与铜金属相互作用过程中电子与晶格耦合项可以忽略,而传导项不可忽略。求解适当激光功率下的双温方程,得到了激光作用中心电子与晶格在不同脉冲宽度激光辐照下的温度变化关系。根据100 fs激光作用后晶格温度场的空间分布情况,研究了激光作用的相分离区域、相爆炸区域以及熔融区域的分布情况。     

Femtosecond laser-induced stimulation of a single neuron in a neuronal network

We demonstrated the stimulation of neurons at a single-cell level in cultured neuronal network by a focused femtosecond laser. When the femtosecond laser was focused on a neuron loaded with a fluorescent calcium indicator, the fluorescence intensity immediately increased at the laser spot, suggesting that intracellular Ca²⁺ increases in the neuronal cell due to the femtosecond laser irradiation. The probability of Ca²⁺ elevation at the laser spot depended on the average laser power, irradiation time, and position of the focal point along the optical axis, indicating that the femtosecond laser activates neurons because of multiphoton absorption. Moreover, after laser irradiation of a single neuron cultured on multielectrode arrays, the evoked electrical activity of the neurons was demonstrated by electrophysiological systems, which concluded that the focused femtosecond laser could achieve stimulating a single neuron in a neuronal network with high spatial and temporal resolution.     

单脉冲飞秒激光时域参数测量技术研究

钛宝石飞秒激光放大系统具有重复频率低、脉冲波形复杂等特点,为准确测量其峰值功率,提出对单次脉冲波形和脉冲宽度测量的需求。介绍了单脉冲飞秒激光时域波形和脉冲宽度的测量原理和测量方法,设计了基于频率分辨光学开关法的单脉冲飞秒激光时域参数测量装置。讨论了单脉冲飞秒激光时域参数测量校准面临的问题及解决方法。     

Control of the photodissociation of CsCl

A femtosecond UV laser pulse is used to resonantly excite CsCl molecules from the ionically bound ground state to the first excited repulsive state. The excitation leads to the dissociation of CsCl. After a certain time delay a visible (VIS) femtosecond laser pulse interrupts the dissociation process by resonantly de-exciting the molecule back to the ground state. According to the Tannor–Rice control scheme, the fraction of dissociated CsCl molecules is controlled by changing the delay time between the two fs laser pulses. The processes involved are investigated theoretically and experimentally. Based on the results, a self-learning system has been realized, which is able to control the dissociation without any a priori knowledge of the molecule. Received: 2 December 1999 / Published online: 24 July 2000     

基于多光子脉冲内干涉相位扫描法对飞秒激光脉冲进行相位测量和补偿的研究

研制了一套基于多光子脉冲内干涉相位扫描方法的可以同时对飞秒激光脉冲进行相位测量和补偿的实验系统装置.实验中,通过自主研发的LabVIEW程序控制液晶空间光调制器和光纤光谱仪,对待测飞秒激光脉冲施加相位扫描,并同时记录受到调制的飞秒激光脉冲的倍频光谱,得到了多光子脉冲内干涉相位扫描(MIIPS)轨迹图.通过MIIPS轨迹图的三次测量和迭代运算,还原出了经过预先啁啾调制的中心波长约为810 nm、重复频率为1 kHz的飞秒激光脉冲的光谱相位,测量精度在0.1 rad以内.根据测量结果,利用液晶空间光调制器对该飞秒激光脉冲进行相位补偿,得到了近似傅里叶变换极限的飞秒激光脉冲.这一装置将在多光子显微成像、脉冲整形、飞秒激光光谱学等众多领域发挥重要作用.     

Removal of doped poly(methylmetacrylate) from tungsten and titanium substrates by femto- and nanosecond laser cleaning

The influence of different laser pulse lengths on the removal of a polymer layer from metal substrates was investigated. As model systems, doped poly(methylmetacrylate) (PMMA) on titanium and tungsten substrates were selected.The ablation threshold and irradiation spot morphology of titanium and tungsten were compared for femtosecond (fs) and nanosecond (ns) laser irradiation and different pulse numbers. Nanosecond laser treatment resulted in a non-homogeneous surface morphology for both titanium and tungsten substrates. Femtosecond irradiation of tungsten revealed a homogeneous ablation spot with little changes in the surface morphology. For titanium, the formation of columnar structures within the irradiation spot was observed.Two different dopant concentrations were used for PMMA to achieve an equal linear absorption coefficient for the femto- and nanosecond laser wavelengths of 790 and 1064 nm. The best results were achieved for the removal of doped PMMA by femtosecond laser irradiation, where only a minimal modification of the metal surface was detected. In the case of nanosecond laser exposure, a pronounced change of the structure was observed, suggesting that damage-free cleaning of the selected metal may only be possible using femtosecond laser pulses. Different experimental parameters, such as laser fluence, pulse repetition rate and sample speed were also investigated to optimize the cleaning quality of doped PMMA from tungsten substrates with femtosecond laser pulses.