基于受激辐射信号的谱域光学相干层析分子成像方法

鉴于不同生理病理状态下组织复折射率实部的变化不大,传统光学相干层析(OCT)成像技术在分子特异性识别上存在先天不足.为此,本文提出了基于受激辐射信号的OCT成像方法,可在实现传统散射成像的同时,实现基于受激辐射信号的分子成像.在超高分辨率谱域OCT系统的基础上,通过增设光谱分光与调制抽运光支路,建立了基于单宽谱光源的抽运探测谱域OCT系统,详细推导了调制抽运下受激辐射信号的获取与成像公式.利用搭建的抽运探测谱域OCT系统,实现了瞬态受激辐射信号的相干探测.基于同时获取的受激辐射OCT信号和传统OCT信号,成功重构了氮化物粉末构建样品的基于受激辐射信号的分子对比OCT图像.     

Spectral domain second-harmonic optical coherence tomography

Optical coherence tomography (OCT) provides micrometer-scale structural imaging by coherent detection of backscattered light. Molecular contrast in OCT has been demonstrated using transient absorption, coherent anti-Stokes Raman scattering, and second-harmonic (SH) generation. The sensitivity of molecular contrast signals can be enhanced by use of Fourier domain techniques. We have constructed a spectrometer-based Fourier domain SH-OCT system for simultaneous acquisition of the fundamental and SH signals. We report a >30 dB increase in SH sensitivity over a similar time domain SH-OCT system and demonstrate contrast between cartilage and bone using collagen as the contrast agent.     

基于光热效应的显微光谱技术在单粒子检测中应用和发展

高灵敏度的单粒子检测技术是纳米粒子在生物医学、化学、光电子等领域应用的前提条件。常见的单粒子检测技术主要包括基于粒子的荧光、拉曼、散射和吸收等信号而发展起来的光学显微成像及光谱技术。其中,拉曼光谱和荧光光谱技术主要适用于一些具有拉曼活性的分子/粒子或可发光的荧光分子或粒子,然而即使对于荧光效率高的有机染料分子和半导体纳米粒子,固有的光漂白和blinking现象也对单粒子探测形成了挑战。散射光谱测量是应用于单粒子检测的另外一种方法,从理论上讲,由于瑞利散射随着尺寸的减小而呈六次方减弱的趋势,在细胞或生物组织内,小尺寸粒子的散射信号很难从背景散射噪声中分离出来。众所周知,介质吸收激发光后会引起介质内的折射率变化,进而在光加热区附近出现折射率的梯度分布,称为光热效应(photothermal effect)。基于粒子光热效应的光学显微成像和光谱测量技术具有信号灵敏度高、无背景散射、原位和免标记等优点,在单粒子检测领域展现了良好的应用潜力。综述了近年来基于光热效应的显微光谱技术在单粒子检测中应用和研究发展,首先介绍了光热效应的测量原理;接着分别讨论了光热透镜测量技术、微分干涉相差测量技术和光热外差测量技术的实验装置,比较了各种测量技术的信噪比、灵敏度、分辨率等特点,并且介绍这些测量技术在单粒子检测中的应用研究进展;接着,论述了近年来研究人员在提高光热显微测量的信噪比、改善动态测量性能以及在红外波段拓展等方面的最新研究成果;最后,简单总结了光热测量技术在单粒子检测领域所面临的挑战。     

聚吡咯修饰的金纳米棒对肿瘤的光学相干层析成像的影响

聚吡咯(PPy)制备简单、生物相容性好,且在近红外(NIR)光谱范围内有很强的吸收,可作为一种良好的光热治疗试剂;同时,其NIR光吸收性质也可用于增强光学相干层析成像(OCT)的对比效果。因此,采用PPy对传统的OCT对比试剂——金纳米棒(GNR)进行表面修饰,有望获得对比效果更好且生物毒性较小的新型OCT对比试剂。选用吡咯为起始原料,在GNR表面进行一步简单的氧化聚合反应即可制备得到PPy修饰的金纳米棒(GNR-PPy)。利用紫外-可见吸收光谱,拉曼光谱和透射电子显微镜对制备的样品进行了分析和表征。构建小鼠荷瘤模型,以研究GNR-PPy对肿瘤OCT图像对比度的增强效果。采用中心波长为840 nm的OCT系统对注射了纳米粒子的肿瘤区域进行OCT成像。结果表明,肿瘤组织注射了GNR-PPy后,OCT信号衰减非常明显;与注射了GNR的OCT图像相比,840 nm光在GNR-PPy的OCT图像中的穿透深度明显更低。从OCT图像中抽提出一维的衰减曲线对OCT图像进行定量分析,发现注射有GNR-PPy肿瘤组织的OCT信号衰减系数明显高于注射了GNR的组织。表明,相对于GNR,GNR-PPy具有更好的OCT信号对比效果,这在增强肿瘤成像效果方面具有潜在应用价值。     

复谱频域OCT成像的研究

采用频域技术的OCT系统，深度扫描信息由背向散射光谱的傅立叶反变换获得，简化了轴向扫描过程，从而使快速OCT成像成为可能．为了实现复杂生物组织的OCT快速成像，消除谱频域OCT的“混叠”现象、重建样品的真实层析结构，本文引入了相移干涉技术，构建了一套满量程复谱频域OCT实验系统，并为系统设计了特殊的分束镜和移相器，为最终实现复杂生物组织的OCT快速成像提供了条件．     

Time-gated Fourier-domain optical coherence tomography

A novel optical coherence tomography (OCT) system is presented that combines Fourier-domain OCT with incoherent nonlinear time gating. By processing backscattered light in the optical domain, the user can select a restricted depth field of view for improved contrast and acquisition speed. This technique has the additional advantage that imaging is done in the infrared (approximately 1280 nm) but is detected in the visible(approximately 504 nm).     

Principles and calibration of collinear photofragmentation and atomic absorption spectroscopy

The kinetics of signal formation in collinear photofragmentation and atomic absorption spectroscopy (CPFAAS) are discussed, and theoretical equations describing the relation between the concentration of the target molecule and the detected atomic absorption in case of pure and impure samples are derived. The validity of the equation for pure samples is studied experimentally by comparing measured target molecule concentrations to concentrations determined using two other independent techniques. Our study shows that CPFAAS is capable of measuring target molecule concentrations from parts per billion (ppb) to hundreds of parts per million (ppm) in microsecond timescale. Moreover, the possibility to extend the dynamic range to cover eight orders of magnitude with a proper selection of fragmentation light source is discussed. The maximum deviation between the CPFAAS technique and a reference measurement technique is found to be less than 5 %. In this study, potassium chloride vapor and atomic potassium are used as a target molecule and a probed atom, respectively.     

Correlation characteristics of optical coherence tomography images of turbid media with statistically inhomogeneous optical parameters

Noisy structure of optical coherence tomography (OCT) images of turbid medium contains information about spatial variations of its optical parameters. We propose analytical model of statistical characteristics of OCT signal fluctuations from turbid medium with spatially inhomogeneous coefficients of absorption and backscattering. Analytically predicted correlation characteristics of OCT signal from spatially inhomogeneous medium are in good agreement with the results of correlation analysis of OCT images of different biological tissues. The proposed model can be efficiently applied for quantitative evaluation of statistical properties of absorption and backscattering fluctuations basing on correlation characteristics of OCT images.     

Theory of laser-induced amplitude and phase gratings including photoselection,orientational relaxation and population kinetics

The generation of amplitude and phase gratings in an absorbing medium is discussed, including the effects of light polarization, population kinetics and orientational relaxation. A theory is given which describes light diffraction by a combination of an amplitude and a phase grating. It is shown that the coherent coupling effect in transient absorption measurements is solely due to the amplitude grating. Weak absorption and a low triplet yield favour diffraction of the probing beam by the phase grating in triple-beam experiments. Phase grating experiments can be performed with highly soluble absorber dyes, the absorption maximum of which is far away from the laser wavelength. Due to photoselection, diffraction by an amplitude grating depends on the polarization of the probing beam while phase grating diffraction is insensitive to this. Perpendicular polarization of the two exciting pulses prevents the buildup of a phase grating and results in light diffracted by the amplitude grating that is proportional to the transient dichroism signal as known from the common absorption relaxation.     

正交双路光纤电流互感器的信号处理方法

利用正交双路光纤传输光电流信号的处理方法,满足了光纤电流互感器在电力系统暂态保护应用中的实时处理要求.此方法可从正交双路光信号中直接提取两路电流信息,减小了光路传输不理想对电流测量值的影响.结合模数变换过采样技术和先进的数字信号处理技术,提高了系统对暂态故障的响应速度,改进了检测性能.实验表明,此方法在暂态信号的采集和实时处理方面,比现有方法的迟延小且精度高.     

Molecularly sensitive optical coherence tomography

Molecular contrast in optical coherence tomography (OCT) is demonstrated by use of coherent anti-Stokes Raman scattering (CARS) for molecular sensitivity. Femtosecond laser pulses are focused into a sample by use of a low-numerical-aperture lens to generate CARS photons, and the backreflected CARS signal is interferometrically measured. With the chemical selectivity provided by CARS and the advanced imaging capabilities of OCT, this technique may be useful for molecular contrast imaging in biological tissues. CARS can be generated and interferometrically measured over at least 600 microm of the depth of field of a low-numerical-aperture objective.     

Protein-based molecular contrast optical coherence tomography with phytochrome as the contrast agent

We report the use of phytochrome A (phyA), a plant protein that can reversibly switch between two states with different absorption maxima (at 660 and 730 nm), as a contrast agent for molecular contrast optical coherence tomography (MCOCT). Our MCOCT scheme builds up a difference image revealing the distribution of phyA within a target sample from pairs of consecutive OCT A-scans acquired at a probe wavelength of 750 nm, both with and without additional illumination of the target sample with 660-nm light. We demonstrate molecular imaging with this new MCOCT modality in a target sample containing a mixture of 0.2% Intralipid and 83 microM of phyA.     

Optical Coherence Tomography: its role in the non-invasive structural examination and conservation of cultural heritage objects—a review

A brief introduction to Optical Coherence Tomography (OCT) is presented, stressing the origin of the tomographic signal and the detection methods defining various modalities of the technique. The parameters of the tomographs, such as axial and lateral resolution, wavelength and intensity of the probing light, imaging range, time of examination, and sensitivity are then defined, and a paradigm for interpreting the OCT tomograms provided. The second part of the article comprises a review of the utilisation of OCT for structural examination of artworks, illustrated with some representative results. Applications to the structural imaging of semi-transparent subsurface layers such as varnishes and glazes, of underdrawings and of reverse painting on glass, are described first, and then applications in the examination of the structure and state of preservation of historic glass, jade, glazed porcelain and faience are discussed. Finally, the use of OCT combined with LIBS analysis and laser ablation of surface layers is presented.     

Simulation of basic characteristics of single-shot full-field optical coherence tomography using spatially phase-modulated reference light

For the single-shot full-field optical coherence tomography (OCT) using spatially phase-modulated reference light, the basic characteristics have been simulated. At low spatial frequencies, the OCT signal intensity is enhanced twofold owing to subtractions, and with increasing the spatial frequency, the OCT signal intensity decreases 0.636 times at half the Nyquist frequency. OCT signal intensities also depend on orientations in images. Residual noninterference components of signal intensities between adjacent uniform areas increase background noise and reduce the system sensitivity. In the reference light, the optimum phase difference between adjacent uniform areas is 180 deg. Deviations from 180 deg reduce subtracted interference components. It is important that interference intensity, noninterference components and phases between adjacent uniform areas be approximately the same to obtain the OCT image with the reduction of background noise stably.     

四能级原子量子相干过程中瞬态光学特性

利用半经典理论研究了N-型四能级相干原子的瞬态光学特性。理论分析表明：当信号光开启的瞬间，介质对探针光的非线性吸收从接近于零的状态迅速增加，经过驰豫振荡过程，趋于稳定状态。本文的研究结果将为吸收型光子开关的设计提供理论依据和参考。     

Speckle variance detection of microvasculature using swept-source optical coherence tomography

We report on imaging of microcirculation by calculating the speckle variance of optical coherence tomography (OCT) structural images acquired using a Fourier domain mode-locked swept-wavelength laser. The algorithm calculates interframe speckle variance in two-dimensional and three-dimensional OCT data sets and shows little dependence to the Doppler angle ranging from 75 degrees to 90 degrees . We demonstrate in vivo detection of blood flow in vessels as small as 25 microm in diameter in a dorsal skinfold window chamber model with direct comparison with intravital fluorescence confocal microscopy. This technique can visualize vessel-size-dependent vascular shutdown and transient vascular occlusion during Visudyne photodynamic therapy and may provide opportunities for studying therapeutic effects of antivascular treatments without on exogenous contrast agent.     

瞬态吸收光谱茜素超快动力学研究

激发态质子转移是光物理学、光化学和光生物过程中最基本的化学反应之一。激发态分子内质子转移(excited-state intramolecular proton transfer, ESIPT)通常是指有机分子受到激发,到达激发态后,质子在激发态势能面上从质子供体基团转移到质子受体基团并形成含有分子内氢键多元环的过程, 一般发生在亚皮秒量级。质子转移可应用于有机发光二极管、荧光探针等领域。茜素,即1,2-二羟基蒽醌,可从茜草根部提取,具有与醌类衍生物相似的结构,常用于染料、染色剂和药物等。近年来,发现茜素分子具有质子转移特性,可用来制备新型“绿色”染料敏化电池。利用稳态吸收、稳态荧光和飞秒瞬态吸收光谱技术以及第一性原理理论计算对溶于乙醇溶液的茜素分子的质子转移过程进行了研究和分析。稳态吸收和稳态荧光研究结果表明: 在基态时,茜素分子的正常构型9,10-酮处于稳定状态,容易发生跃迁;在激发态时,茜素分子的互变异构体构型1,10-酮处于稳定状态,容易产生荧光发射。飞秒瞬态吸收光谱测量使用的激光的激发波长为370 nm。测得的瞬态吸收光谱在430 nm附近存在茜素的基态漂白信号。通过使用全局拟合方法对瞬态吸收光谱进行分析研究发现：茜素正常构型9,10-酮的激发态分子内质子转移时间为110.5 fs,茜素互变异构体构型1,10-酮分子内振动弛豫时间为30.7 ps,茜素互变异构体构型1,10-酮荧光寿命为131.7 ps。通过使用单波长动力学拟合的方法对瞬态吸收光谱进行分析发现：发生质子转移的时间尺度与运用全局拟合方法得出的结果基本一致;茜素分子的正常构型9,10-酮分子在110.5 fs的时间尺度内处于快速减少的趋势,而茜素分子的互变异构体构型1,10-酮分子在这一时间尺度内处于快速上升的趋势。当延迟时间增大时,茜素分子的互变异构体构型1,10-酮分子又呈现缓慢衰减的趋势。     

激光激发等离子体中的光吸收实验

利用两个激光等离子体相互作用获得离子的吸收光谱的技术已得到很大改善。实验中采用了新的信号检测系统和新的数据处理技术。此实验技术已成功地应用到对瞬态的中等Z离子的吸收谱的测量。获得了BeⅣ、BeⅢ、BeⅡ和BeⅠ离子的1s电子的吸收谱及相关的软X射线范围内的光电离谱。简要地实验得到的碳离子的初步结果。     

超快微光分子光谱探测技术研究

按照超快极微弱分子吸收光谱学,荧光光谱学,时间分辨光谱学以及偏振荧光光谱学探测特性,设计、集成组建了能够探测紫外－可见－红外快到飞秒时间分辨的单分子光学事件的激光瞬态光谱仪.其光源有从300 nm到3000 nm连续可调的飞秒激光器、纳秒氢灯及连续氙灯.光谱分辨率达0.05 nm,在泵浦探测差异吸收下有小于150 fs的时间分辨率.谱仪能够实时给出光谱曲线及生物分子组分寿命.利用该谱仪探测了PSⅡCC,PSⅡRC的能量传递动力学.在83 K温度下PSⅡCC中的β-Car分子接收507 nm光能,以单步跃迁和随机转移的方式通过Chl a641.5637/638分子传递光能到反应中心Chl a683.2680/681,平均传能时间为77 ps,有59.94％的组分用355 ps时间电荷重组.在PSⅡRC中的β-Car分子接收507 nm光能,由Chl a641.5637/638分子传递光能通过Chl a678.2675.5到反应中心,平均传能时间为88.5 ps.在83 K温度下,反应中心复合物离子对;P680+·pheo-］平均再复合寿命为19.35 ns.     

Spectroscopic optical coherence tomography

Spectroscopic optical coherence tomography (OCT), an extension of conventional OCT, is demonstrated for performing cross-sectional tomographic and spectroscopic imaging. Information on the spectral content of backscattered light is obtained by detection and processing of the interferometric OCT signal. This method allows the spectrum of backscattered light to be measured over the entire available optical bandwidth simultaneously in a single measurement. Specific spectral features can be extracted by use of digital signal processing without changing the measurement apparatus. An ultrabroadband femtosecond Ti:Al(2)O(3) laser was used to achieve spectroscopic imaging over the wavelength range from 650 to 1000 nm in a simple model as well as in vivo in the Xenopus laevis (African frog) tadpole. Multidimensional spectroscopic data are displayed by use of a novel hue-saturation false-color mapping.