大空间生物组织透射的Monte Carlo模拟方法的加速

乳腺等类型生物组织的透射Monte Carlo模拟(简称MC模拟)耗时很长,这主要是由于被模拟的组织较厚和Monte Carlo模拟自身统计特性引起的所需光子数量多造成的。针对光源与接收面在同一直线上,采用光纤接收透射光的模型进行分析,通过考虑生物组织自身的光学特性,进行了边界约束和有效后向散射次数的限制来缩短仿真时间,经过多次实验验证,以两次散射所到位置为分界,并充分利用组织光学参数计算约束边界,计算相对比较简单,同时考虑到入射与出射的实际情况进行微扩,从而得到的约束边界。对光学参数相同但厚度不同的组织进行仿真,分析进入同一位置光纤的光子数所发生后向散射次数,发现后向散射次数随着组织厚度与散射系数的增大而增大,随着吸收系数与各向异性因子的增大而减小,通过限定后向散射次数来节省时间。仿真结果表明,在传统计算机上,该方法能够显著缩短MC模拟的运算时间50%左右,尤其适用于较厚且边界较为复杂的高散射物质的MC模拟。该提速方法主要应用于光源与接收面处于同一直线,且生物组织较厚,散射系数较大的情况,能够有效节约仿真所需时间,有利于分析组织透射成像。     

Noninvasive determination of tissue optical properties based on radiative transfer theory

We present a feasibility study of a new method for determining the tissue optical properties, including the absorption and scattering coefficients and the scattering asymmetry factor. A state-of-the-art radiative transfer model for the coupled air/tissue system, based on rigorous radiative transfer theory, is used in our forward modeling simulations. The concept of the effective photon penetration depth is introduced and used to help determine the depth below, which information about the tissue will not be available through noninvasive imaging of a biological tissue using reflected diffuse light. Simulation results show that for accurate determination of tissue optical properties, one can use radiative transfer theory in conjunction with measurements of reflected radiances as well as other existing techniques.     

Graphical thresholding procedure and optimal light level estimation for spatially resolved photon counting with EMCCDs

Thanks to the electron multiplying function that can effectively convert the weak incident photon signal to amplified electron output, electron multiplying charged-coupled devices (EMCCDs) are becoming useful and popular detectors in photon counting regimes necessitating also spatial resolution. A multi-imaging strategy has been already proposed and experimentally tested to improve the accuracy of photon counting with an EMCCD, by taking into account the random nature of its on-chip gain and the possibility of multiple photodetection events on 1 pixel. In this paper, referring to the thresholding procedure developed for photon counting, we present a clear graphical method for the threshold and the optimal light level estimations. Thanks to the graphical visualization of the probabilities involved in the detection errors on 1 pixel, we are able to derive in a straightforward way and for any EMCCD, the threshold level and thus the best mean level of illumination to be used in order to minimize the false detection probabilities that might ruin the image statistics, especially in cases where quantum spatial effects might be observed.     

Diffuse optical imaging and spectroscopy,in vivo

Based on photon migration the new goal of diffuse optical imaging is to reveal optical contrasts in the depth of biological tissues. We discuss first the origin of contrast mechanism (absorption, fluorescence and scattering) used on diffuse optical imaging and spectroscopy. Then, various experimental approaches are described based on CW, pulsed and modulated light excitation and detection. Theoretical models which provide solutions for direct and inverse problems are presented using random walk theory. Finally two studies on breast imaging and on the use of fluorescence exogeneous markers are discussed in detail.     

基于蒙特卡罗模拟的散射介质中后向光散射模型及分析应用

构建了内光源模型探讨散射介质中的光散射现象,利用蒙特卡罗方法研究了逃逸出组织的后向散射光子数随光子在组织内部发生的散射次数的分布关系,探讨了光源照明方式、辐射强度、接收方式、调制等参数的变化对后向散射的影响,结果表明后向散射光子的数量随散射次数的分布并非简单的单调递增或递减,而是一条先增大后减小出现峰值的曲线. 峰值位置、峰值大小及曲线形状与光源、探测方式、组织光学特性参数等有关. 关键词： 医用光学与生物技术 散射介质 后向散射 蒙特卡罗模拟     

利用多像素光子计数器的光学串话效应提高光子探测效率

通过分析多像素光子计数器(MPPC)的工作原理和其光学串话(OC)效应的特点,提出在使用MPPC输出雪崩信号的幅度或电荷量作为光子计数的参量时,利用MPPC的OC效应能提高MPPC的光子探测效率的观点,并从理论上分析了OC效应对光子探测效率的影响。理论分析结果显示,在这两种光子计数模式下,利用OC效应能明显提高MPPC的光子探测效率。利用本文模型计算得出当MPPC的雪崩单元数M为1 600个,忽略OC效应时的光子探测效率等于30%,光学串话概率等于50%,以及单脉冲入射光子数均值为10时,包含OC效应影响的等效光子探测效率可提高50%,达45%左右。该结果对MPPC在天体物理、粒子物理、荧光光谱探测等弱光探测场合的应用有一定指导意义。     

Experimental proof of the feasibility of using an angled fiber-optic probe for depth-sensitive fluorescence spectroscopy of turbid media

An angled fiber-optic probe that facilitates depth-sensitive fluorescence measurements was developed for enhancing detection of epithelial precancers. The probe was tested on solid, two-layered phantoms and proved to be effective in selectively detecting fluorescence from different layers. Specifically, a larger illumination angle provides greater sensitivity to fluorescence from the top layer as well as yielding an overall higher fluorescence signal. Monte Carlo simulations of a theoretical model of the phantoms demonstrate that increasing the illumination angle results in an increased excitation photon density and, thus, in increased fluorescence generated in the top layer.     

Dark-field optical coherence microscopy

Dark-field illumination is known to enhance scattering contrast in optical microscopy. We combined this concept with Fourier domain optical coherence microscopy (OCM). The detection and illumination paths are decoupled, and only the scattered light originating from the sample generates the tomogram signal, whereas any specular reflection is highly suppressed. We analyze and discuss this dark-field OCM concept and present its superior imaging quality on live cell samples.     

Performance analysis of a swept-source optical coherence tomography system with a quadrature interferometer and optical amplification

A performance analysis of signal to noise ratio for an optical coherence tomography system with quadrature detection and a semiconductor optical amplifier in the sample arm is discussed. The results are compared and discussed in relation to a conventional OCT system (without optical amplification). An increase of the signal to noise ratio up to 14 dB at a depth of 0.5 mm is obtained compared to the system without the optical amplifier. Overall, an improvement was demonstrated for signal coming from deeper regions within the samples. Arterial plaque from a myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbit is visualized and characterized using this system. Improvement of signal to noise ratio increases the penetration depth possible for OCT images, from 1 mm to 2 mm within the vessel wall of an artery. Preliminary results show that vulnerable plaque with fibrous cap, macrophage accumulations and calcification in the arterial tissue are measurable with this OCT system.     

Low cost optical detection of DNA hybridization on biochips

The detection of DNA hybridization can be either complicated or require expensive devices. Some methods use a fluorescence signal to investigate hybridization. Here, we present an optical probe based on optical fibers both for illumination and for fluorescence collection. The detection is made with a microspectrophotometer and the signal is then treated with a PC. We also developed the DNA biochips (glass plate with a gold layer functionalized with target DNA) as well as a dedicated microtank which maintains a constant flow of probe DNA over the target one. Finally, the detection of hybridization with this simple and versatile system is presented.     

Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations

We report on measurements and Monte Carlo analysis of the spatial distribution of light in two-photon excitation fluorescence (TPEF) microscopy operating through scattering media. The axial profile of TPEF produced by a high numerical aperture microscope objective focusing through a tissue-like optical phantom at depths more than five photon mean-scattering lengths is studied. The measured profile is quantitatively interpreted with time-resolved Monte Carlo simulations that take into account the spatio-temporal distribution of the photons within the turbid medium under a femtosecond excitation regime. It is shown that considering only the ballistic photons leads to an under-estimation of the actual out-of-focus distribution of the fluorescence, whereas it is over-estimated when the ballistic and scattered photons are treated alike. Comparison of the in-focus signal to the overall out-of-focus background clearly pointed out that the signal-to-background ratio of TPEF microscopy images benefits from a shortening of the excitation pulses. Moreover, in homogeneously labeled specimen, the background is shown to overcome the signal at imaging depths of about five to six photon mean-scattering paths.     

Enhancement of electron spin coherence by optical preparation of nuclear spins

We study a large ensemble of nuclear spins interacting with a single electron spin in a quantum dot under optical excitation and photon detection. At the two-photon resonance between the two electron-spin states, the detection of light scattering from the intermediate exciton state acts as a weak quantum measurement of the effective magnetic (Overhauser) field due to the nuclear spins. In a coherent population trapping state without light scattering, the nuclear state is projected into an eigenstate of the Overhauser field operator, and electron decoherence due to nuclear spins is suppressed: We show that this limit can be approached by adapting the driving frequencies when a photon is detected. We use a Lindblad equation to describe the driven system under photon emission and detection. Numerically, we find an increase of the electron coherence time from 5 to 500 ns after a preparation time of 10 micros.     

Spatial Fourier-decomposition optical fluorescen tomography-theoretical investigation

A new three-dimensional (3D) optical fluorescent tomographic imaging scheme is proposed with structured illumination and spatial Fourierdomain decomposition methods for the first time. In this spatial Fourier-decomposition optical fluorescence tomography (SF-OFT), the intensity of focused excitation light from an objective lens is modulated to be a cosine function along the optical axis of the system. For a given position in a two-dimensional (2D) raster scanning process, the spatial frequency of the cosine function along the optical axis sweeps in a proper range while a series of fluorescence intensity are detected accordingly. By making an inverse discrete cosine transformation of these recorded intensity profiles, the distribution of fluorescent markers along the optical axis of a focused laser beam is obtained. A 3D optical fluorescent tomography can be achieved with this proposed SF-OFT technique with a simple 2D raster scanning process.     

Biochip fluorescence detection system with spatial and spectral separation

A novel optical arrangement for fluorescence detection that employs spatial separation as well as spectral filter to increase the signal to noise ratio is proposed. Using a prism and two mirrors, the elliptical laser beam of a laser diode, as an excitation light, is homogenized and the transmitted excitation light is separated from the fluorescence not to reach the collecting optics. Uncooled CCD can capture the fluorescence image of up to 40 fluorescently-labeled protein patterns without scanning or mechanical translation. This paper presents the simulation, construction and measurement results of the developed optical system. The measurements show that the combination of prism and mirrors converts the excitation light from the laser diode to uniform illumination on the specimen, and provides the separation between excitation and fluorescence light to give high signal to noise ratio. It is also possible to assay various protein concentrations ranging from 1000 to 10 ng/ml reliably. We believe that the proposed fluorescence detection system can be used to build a commercially valuable, low cost, hand-held or miniature fast detection device for point-of-care applications.     

Analysis of influence of black matrix effect in spatial light modulator on optical correlation detection

Motion detection with a joint transform optical correlator is based on the maximally correlated bright spots in a correlation image; therefore, the quality of a correlation image greatly influences optical correlation detection. As the core component of a joint transform optical correlator, a spatial light modulator has an opaque part (black matrix structure) that is an important factor that needs to be considered. In this paper, we first analyzed the effect of the fill factor on the light energy distribution in the image plane according to the mechanism of the spatial light modulator using a multiple-subpixel matrix to simulate a single pixel, while zeroing some subpixels to simulate the black matrix part in the single pixel and employing computer software to simulate the joint transform optical correlator to obtain the simulated correlation image result with a black matrix effect. In addition, we built an experimental setup to obtain an actually photographed correlation image, which was well consistent with the simulated result.     

应用于数字高速成像的离轴光学系统设计

为提高数字高速多幅相机的像面照度,简化光学系统结构,提出一种离轴光学系统设计方法。该系统使用多组相同物镜,采用圆心一组,圆周均匀分布多组的方式平行阵列排布。分布于圆周上的物镜,其像面接收器相对物镜轴心平移离轴,以获得与圆心物镜相同的物面图像。分析了单物镜像面照度、畸变等与视场有关参数对光学系统各像面匹配的影响。在此基础上给出一个5通道离轴光学系统设计实例,系统空间分辨率优于30 lp/mm,畸变小于0.1%,F数2.0。相比目前广泛采用的棱锥、棱镜分幅方式,该离轴光学系统像面照度增加5倍以上,系统结构简易。     

On the Emission Intensity of Fluorescent Microspheres in Cardiac Tissue Images

Formulations for the total fluorescence intensity of fluorescent microspheres in slabs of cardiac tissue were determined experimentally and theoretically. The tissue depth, at which the slab can be considered as a semi-infinite turbid medium, and critical layer thickness, which accounts for the most emission intensity were evaluated to be 8–9 and 3–5 mm, respectively, for the cardiac tissue. When fluorescent microspheres are linearly distributed across the slab depth, the mean absorption of them is proportional to the sum of their normalized total emissions in the slab excited from both sides. The formulations may be used for the fluorescence images analysis of cardiac and other biological tissues.     

Integrated optical microcavities for enhanced evanescent-wave spectroscopy

The use of integrated optical microcavities (MCs) for enhanced optical spectroscopy and sensing is investigated. The MC sustains high- Q whispering-gallery modes, in which the energy of the optical field can be efficiently stored. The resulting enhanced field can be used to probe fluorescent molecules in the cladding of the MC. Enhanced fluorescence excitation with an integrated optical MC is demonstrated experimentally for what is believed to be the first time. A comparison between a MC and a straight waveguide shows that the MC may give an increase of 40 times in fluorescence excitation. Because of the ultrasmall size of the MC (15 microm in radius), the fluorescence signal may be observed from only 20 molecules in the cladding.     

Vacuum-stimulated raman scattering based on adiabatic passage in a high-finesse optical cavity

We report on the first observation of stimulated Raman scattering from a Lambda-type three-level atom, where the stimulation is realized by the vacuum field of a high-finesse optical cavity. The scheme produces one intracavity photon by means of an adiabatic passage technique based on a counterintuitive interaction sequence between pump laser and cavity field. This photon leaves the cavity through the less-reflecting mirror. The emission rate shows a characteristic dependence on the cavity and pump detuning, and the observed spectra have a subnatural linewidth. The results are in excellent agreement with numerical simulations.     

Imaging optically scattering objects with ultrasound-modulated optical tomography

We show the feasibility of imaging objects having different optical scattering coefficients relative to the surrounding scattering medium using ultrasound-modulated optical tomography (UOT). While the spatial resolution depends on ultrasound parameters, the image contrast depends on the difference in scattering coefficient between the object and the surrounding medium. Experimental measurements obtained with a CCD-based speckle contrast detection scheme are in agreement with Monte Carlo simulations and analytical calculations. This study complements previous UOT experiments that demonstrated optical absorption contrast.