相干反斯托克斯拉曼散射显微镜的成像特性

对自身不发荧光且不便于荧光标记的化学或生物学样品，集相干反斯托克斯拉曼散射与激光共焦扫描显微镜于一身的相干反斯托克斯拉曼散射显微镜是一种好的选择。因为相干反斯托克斯拉曼散射是一种非线性过程，相干反斯托克斯拉曼散射显微镜的显微成像特性与一般的共焦显微镜非常不同。首先计算了焦点附近相干反斯托克斯拉曼散射激发场的偏振分布，然后，利用格林函数方法，得到了以赫兹偶极子为源的波动方程的精确解，发现对于不同的成像配置和样品形状，像场的相干反斯托克斯拉曼散射场分布非常不同，因此传统的显微镜成像表征方式（如点扩展函数）将不再能描述相干反斯托克斯拉曼散射显微镜的成像特性。     

超衍射极限相干反斯托克斯拉曼散射显微成像技术及其探测极限分析

理论上提出一种突破衍射极限限制的相干反斯托克斯拉曼散射显微成像方法, 并对其探测极限进行分析.通过引入环形附加探测光与艾里斑周边的声子作用, 实现点扩展函数的改造, 提高相干反斯托克斯拉曼散射显微成像系统的横向空间分辨率. 随着分辨率的提高, 信号强度也随之降低, 尤其当应用于生物学、医学研究时, 样品分子数密度通常很低, 这将导致信号探测更加困难. 因此分析系统的探测极限, 确定超分辨体积元内的最小可探测分子数是展开超衍射极限相干反斯 托克斯拉曼散射显微成像实验研究的重要前提. 当泵浦光、斯托克斯光、探测光光强均达到极大值, 分辨率约40 nm三维空间内, 超衍射极限相干反斯托克斯拉曼散射显微成像系统的散粒噪声信噪比由曝 光时间与样品分子数密度决定. 曝光时间若取20 ms, 探测极限约为10³, 样品分子数目只有大于探测极限, 才能保证信号可以从噪声背景中提取出来. 关键词： 突破衍射极限 相干反斯托克斯拉曼散射 非线性光学 探测极限     

基于超连续谱光源的相干反斯托克斯拉曼散射成像理论与实验可行性研究

将超连续谱光纤激光器引入到相干反斯托克斯拉曼散射成像系统,以解决当前相干反斯托克斯拉曼散射成像系统造价昂贵、体积庞大的问题。首先分析了基于超连续谱光源的相干反斯托克斯拉曼散射成像理论及其影响因素;接着搭建了一套基于超连续谱光源的相干反斯托克斯拉曼散射成像系统,并进行了成像性能测试。在物镜、功率、浓度等不同影响因素的情况下,系统测试的实验结果与数值仿真结果具有高度一致性,验证了基于超连续谱光源的相干反斯托克斯拉曼散射成像理论与实验的可行性。     

小尺寸样品CARS图像信号分布实验研究

通过设计实验与分析模型,研究相干反斯托克斯拉曼散射成像对于样品尺寸小于系统点扩展函数尺寸的情况,分析相干反斯托克斯拉曼散射图像周围的depth的形成原因。分析过程首次引入轴向传输动态位移光线（此光线由相干体积元内轴心光线抽象出）对球形或柱形样品直径小于系统点扩展函数横向与轴向尺寸进行建模解析,对于所建模型定量分析结果表明,对于样品尺寸小于系统点扩展函数尺寸样品折射率调制了有效作用深度。Gouy相移只是其表观现象,是一个伴随性特征,因为物理学中存在的另一个极端,当样品尺寸足够小且具有相当的折射率,Gouy相移的作用近似为0,此时,样品周围的depth主要与样品的折射率及系统相干层析体积元内有效作用长度有关,相干反斯托克斯拉曼散射参量信号,具有自身的波矢匹配条件,不仅有大小,有方向,而且还受到样品自身材料折射率的影响比较明显。也就是对于参量过程波矢匹配条件是因,Gouy相移是伴随性特征。自此,通过相干反斯托克斯拉曼散射间接说明所用的紧聚焦参量信号成像过程,正是因为波矢匹配条件的存在使二次信号继承了激发光的紧聚焦特性,而拥有了继承性的Gouy相移,结合实验结果采用物理学的极端假设表明,Gouy相移不是产生depth的主要原因,主要原因是样品及周围的环境的折射率与系统相干层析体积元内有效作用长度之间共同作用的结果。而实验结果与模型定量分析的结果相吻合,此实验帮我们找到了影响CARS图像样品周围depth的成因机制,对于其他小尺寸样品的参量成像结果的分析具有一定的借鉴意义。通过设计实验,结合定量模型分析,首次明确造成小尺寸样品CARS图像周围的depth的真正原因,此抽象模型的拓展性对纳观参量过程的分析具有得天独厚的优势。抽象出的主能量光线动态位移模型成功分析纳观成像结果表明,相干作用长度之内有效作用长度及其行进路径的过程论的主因分析方法是研究纳观作用机制的最佳方法。     

相干反斯托克斯拉曼散射显微成像技术研究

基于全量子理论对相干反斯托克斯拉曼散射(CARS)过程进行了分析, 在此基础上搭建了单频CARS显微成像系统, 获得了不同尺寸聚苯乙烯微球高对比度的CARS显微图像. 为了标定成像系统的空间分辨率, 采用逐点扫描方式对直径为110 nm聚苯乙烯微球成像, 从而重构出系统的点扩展函数. 结果表明: 该CARS显微成像系统的横向空间分辨率约为600 nm, 而由阿贝衍射极限决定的理论空间分辨率约为300 nm. 分析了导致分辨率降低的原因, 并提出了解决方案. 为实现纳米分辨的CARS显微成像打下了坚实的基础.     

基于偏振探测的CARS非共振背景抑制与分离

为消除非共振背景干扰,提高相干反斯托克斯拉曼散射测量精度,研究了基于偏振探测的相干反斯托克斯拉曼散射非共振背景抑制与分离.首先,从理论上证明了相干反斯托克斯拉曼散射信号强度只取决于三阶非线性极化率共振部分,而与非共振部分无关,并且三阶极化率共振部分与非共振部分偏振方向不一致,为偏振相干反斯托克斯拉曼散射提供了理论支撑;然后,以平面火焰炉温度场为研究对象,设计了偏振相干反斯托克斯拉曼散射探测系统实施方案,并与热电偶、常规相干反斯托克斯拉曼散射两种方法在相同条件下开展了温度测量对比实验.实验结果表明:常规相干反斯托克斯拉曼散射方法的测量相对误差优于7%,测量相对不确定度优于7%;偏振相干反斯托克斯拉曼散射方法的测量相对误差优于4%,测量相对不确定度优于5%;相同工况条件下与常规相干反斯托克斯拉曼散射相比,偏振相干反斯托克斯拉曼散射最接近均值的拟合光谱更加平滑.偏振相干反斯托克斯拉曼散射具有更小的拟合误差,是一种更为有效的相干反斯托克斯拉曼散射诊断方法.     

多光子皮肤成像技术及其应用

多光子成像技术是一种层析能力好、信噪比高的新型光学成像技术。在皮肤光学三维检测中,多光子技术已经应用于无创在体成像,且已得到产业化开发。本文将首先介绍多光子皮肤检测系统的若干核心技术,即双光子自发荧光技术、二次谐波成像技术、荧光寿命成像技术、相干反斯托克斯-拉曼成像技术等,然后简要介绍多光子成像系统在皮肤疾病成像检测上的应用,最后分析该系统的优势和未来可能的发展趋势。     

混合溶液中CARS过程的非共振信号实验研究

相干反斯托克斯拉曼散射是一种非线性四波混频效应,但是通过探测共振信号不易进行物质成分的定量光谱分析。本文利用单频相干反斯托克斯拉曼散射光谱分析法,对不同体积比混合的乙醇溶液在远离双光子共振跃迁及其远离溶质与溶剂的特征拉曼共振态位置进行了光谱探测。通过对实验结果进行分析发现,在共振位置2 876 cm-1的信号强度随混合溶液中的乙醇的体积比增加而增加,呈二次方关系。而在远离共振态位置的非共振信号强度随混合溶液中乙醇的体积比增加而增加,且呈线性关系,进而说明了非共振信号强度随着分子数浓度N呈线性变化关系。因此,通过探测非共振信号强度与分子数浓度关系可以为混合物中特定成分的定量光谱分析提供一种研究途径。     

在单模光纤中放大的反斯托克斯拉曼背向自发散射的温度效应

在单模光纤中,输入的激光功率大于阈值时．出现放大的反斯托克斯拉曼背向自发散射现象。实验发现：放大的反斯托克斯拉曼背向自发散射具有温度效应．与反斯托克斯拉曼背向自发散射一样,放大的拉曼散射光的光子通量受到光纤温度的调制。反斯托克斯拉曼背向白发散射的放大效应抑制了单模光纤中的相干噪声,改善了系统的信噪比。实验还发现．放大的反斯托克斯扎曼背向自发散射空域曲线上放大的端点位置随激发功率的增高前移并具有一定的规律性。放大的反斯托克斯拉曼背向自发散射的温度效应作为一种新的测温原理,已应用于远程30km分布光纤温度传感器系统。     

小尺寸样品CARS图像信号分布实验研究（英文）

通过设计实验与分析模型,研究相干反斯托克斯拉曼散射成像对于样品尺寸小于系统点扩展函数尺寸的情况,分析相干反斯托克斯拉曼散射图像周围的depth的形成原因。分析过程首次引入轴向传输动态位移光线(此光线由相干体积元内轴心光线抽象出)对球形或柱形样品直径小于系统点扩展函数横向与轴向尺寸进行建模解析,对于所建模型定量分析结果表明,对于样品尺寸小于系统点扩展函数尺寸样品折射率调制了有效作用深度。Gouy相移只是其表观现象,是一个伴随性特征,因为物理学中存在的另一个极端,当样品尺寸足够小且具有相当的折射率,Gouy相移的作用近似为0,此时,样品周围的depth主要与样品的折射率及系统相干层析体积元内有效作用长度有关,相干反斯托克斯拉曼散射参量信号,具有自身的波矢匹配条件,不仅有大小,有方向,而且还受到样品自身材料折射率的影响比较明显。也就是对于参量过程波矢匹配条件是因,Gouy相移是伴随性特征。自此,通过相干反斯托克斯拉曼散射间接说明所用的紧聚焦参量信号成像过程,正是因为波矢匹配条件的存在使二次信号继承了激发光的紧聚焦特性,而拥有了继承性的Gouy相移,结合实验结果采用物理学的极端假设表明,Gouy相移不是产生depth的主要原因,主要原因是样品及周围的环境的折射率与系统相干层析体积元内有效作用长度之间共同作用的结果。而实验结果与模型定量分析的结果相吻合,此实验帮我们找到了影响CARS图像样品周围depth的成因机制,对于其他小尺寸样品的参量成像结果的分析具有一定的借鉴意义。通过设计实验,结合定量模型分析,首次明确造成小尺寸样品CARS图像周围的depth的真正原因,此抽象模型的拓展性对纳观参量过程的分析具有得天独厚的优势。抽象出的主能量光线动态位移模型成功分析纳观成像结果表明,相干作用长度之内有效作用长度及其行进路径的过程论的主因分析方法是研究纳观作用机制的最佳方法。     

Increasing the imaging depth of coherent anti-Stokes Raman scattering microscopy with a miniature microscope objective

A miniature objective lens with a tip diameter of 1.3 mm was used for extending the penetration depth of coherent anti-Stokes Raman scattering (CARS) microscopy. Its axial and lateral focal widths were determined to be 11.4 and 0.86 microm, respectively, by two-photon excitation fluorescence imaging of 200 nm beads at a 735 nm excitation wavelength. By inserting the lens tip into a soft gel sample, CARS images of 2 microm polystyrene beads 5 mm deep from the surface were acquired. The miniature objective was applied to CARS imaging of rat spinal cord white matter with a minimal requirement for surgery.     

Nonlinear optical properties of [（CH3）4N]Au（dmit）2 using Z-scan technique

The third-order optical nonlinearities of [（CH3）4N]Au（dmit）2 （dmit = 4,5-dithiolate-1,3-dithiole-2-thione） at 532 nm and 1064 nm are investigated using the Z-scan technique with pulses of picoseconds duration. The Z-scan spectra reveal a strong nonlinear absorption （reverse saturable absorption） and a negative nonlinear refraction at 532 nm. No nonlinear absorption is observed at 1064 nm. The molecular second-order hyperpolarizability γ for the [（CH3）4N]Au（dmit）2 molecule at 532nm is estimated to be as high as （2.1 ±0.1） × 10^-31 esu, which is nearly three times larger than that at 1064 nm. The mechanism responsible for the difference between the results is analysed. Nonlinear transmission measurements suggest that this material has potential applications in optical limiting.     

Investigation of third-order nonlinear optical properties of two azo-nickel chelate compounds

This paper investigates the third-order nonlinear optical properties of two azo-nickel chelate compounds by the optical Kerr gate method at 830 nm wavelength with pulse duration of 120 fs. Both of the two compounds exhibited large third-order optical nonlinearity. The second-order hyperpolarizability,γ, of Compound 1 is of 1.0 × 10^-31 esu. Due to the charge transfer, the γ of Compound 2 with electron donor and acceptor group is 4.9 × 10^-31 esu, which is a four-time enhancement in comparison with Compound i. The absorption spectra show that the electron push-pull effect, which induces intramolecular charge transfer, leads to the increased optical nonlinearity.     

Label-free imaging of arterial cells and extracellular matrix using a multimodal CARS microscope

A multimodal nonlinear optical imaging system that integrates coherent anti-Stokes Raman scattering (CARS), sum-frequency generation (SFG), and two-photon excitation fluorescence (TPEF) on the same platform was developed and applied to visualize single cells and extracellular matrix in fresh carotid arteries. CARS signals arising from CH₂-rich membranes allowed visualization of endothelial cells and smooth muscle cells of the arterial wall. Additionally, CARS microscopy allowed vibrational imaging of elastin and collagen fibrils which are also rich in CH₂ bonds. The extracellular matrix organization was further confirmed by TPEF signals arising from elastin’s autofluorescence and SFG signals arising from collagen fibrils’ non-centrosymmetric structure. Label-free imaging of significant components of arterial tissues suggests the potential application of multimodal nonlinear optical microscopy to monitor onset and progression of arterial diseases.     

Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration

We have developed a new coherent anti-Stokes Raman scattering (CARS) microscopy system with a collinear configuration for use in the fingerprint region. The system consists of a picosecond laser system and a transmission-type laser scanning microscope without a pinhole in front of the detector. The observable Raman-shift region is 900-1750 cm(-1), the spectral resolution is 30cm(-1), and the spatial resolution is smaller than 1 mum in the lateral direction and 3.2 mum in the depth direction, with objectives with a numerical aperture of 0.65. CARS spectra and images of polystyrene beads are demonstrated, and CARS imaging of a viable yeast cell is attempted.     

Enhanced optical nonlinear absorption of graded Au--TiO2 composite films

This paper reports that single-layer and graded Au-TiO2 granular composite films with Au atom content 15%- 66% were prepared by using reactive co-sputtering technique. The third-order optical nonlinearity of single-layer and graded composite films was investigated by using s- and p-polarized Z-scans in femtosecond time scale. The nonlinear absorption coefficient βeff of single-layer Au-TiO2 films is measured to be -2.3×10^3-0.76×10^3 cm/GW with Au atom content 15%-66%. The βeff value of the 10-layer Au-TiO2 graded film is enhanced to be -2.1×10^4cm/GW calculated from p-polarized Z-scans, which is about ten times the maximum βeff of single-layer films. Broadened response in the wavelength region 730-860 nm of the enhanced optical nonlinearity of graded Au-TiO2 composite films was also investigated.     

Differential two-signal picosecond-pulse coherent anti-Stokes Raman scattering imaging microscopy by using a dual-mode optical parametric oscillator

We propose and demonstrate a novel differential two-signal technique of coherent anti-Stokes Raman scattering (CARS) imaging microscopy using a picosecond (ps) optical parametric oscillator (OPO). By adjusting a Lyot filter inside the cavity, we operated the OPO oscillating in two stable modes separated by a few nanometers. The CARS images generated by the two modes are separated by a spectrograph behind the microscope setup, and their differential image is directly obtained by balanced lock-in detection. The feasibility of the technique is experimentally verified by imaging micrometer-sized polystyrene beads immersed in water.     

High‐resolution narrowband CARS spectroscopy in the spectral fingerprint region

Coherent anti‐Stokes Raman scattering (CARS) spectroscopy is an important technique for spectroscopy and chemically selective microscopy, but wider implementation requires dedicated versatile tunable sources. We describe an optical parametric oscillator (OPO) based on a magnesium oxide‐doped periodically poled lithium niobate crystal, with a novel variable output coupler, used as a tunable coherent light source. The OPO's signal wavelength ranges from 880 to 1040 nm and its idler wavelength from 1090 to 1350 nm. We use this OPO to demonstrate high‐resolution narrowband CARS spectroscopy on bulk polystyrene from 900 to 3600 cm⁻¹, covering a large part of the molecular fingerprint region. Recording vibrational spectra using narrowband CARS spectroscopy has several advantages over spontaneous Raman spectroscopy, which we discuss. We isolate the resonant part of the CARS spectrum and compare it to the spontaneous Raman spectrum of polystyrene using the maximum entropy method of phase retrieval; we find them to be in extremely good agreement. Copyright © 2009 John Wiley & Sons, Ltd.