Experimental and numerical study on simultaneous effects of scattering and absorption on fluorescence spectroscopy of a breast phantom

Detection of dysplastic lesions can decrease morbidity and mortality caused by cancer. The fluorescence spectroscopy is a noninvasive method of detecting dysplasia in several organs. During dysplastic progression, fluorescence intensity of spectrum is changed due to variation in absorption and scattering coefficients of tissue. In this work we have experimentally verified simultaneous effects of scattering and absorption coefficients on fluorescence intensity of different tissue like phantoms with the same optical properties as the human breast ductal carcinoma. The results are compared with those obtained by Monte Carlo simulation and good agreement between them is observed. This provides an important detecting method to discriminate dysplastic tissue from normal tissue.     

In vivo near-infrared spectral detection of pressure-induced changes in breast tissue

A diffuse near-infrared tomography system was used to measure dynamic changes in the absolute optical properties of the human breast that were induced through pressure applied to the tissue surface. Results from five subjects show that absorption and scattering coefficients changed measurably when pressure was increased and that these relative changes correlated with the subjects' body-mass index, indicating that the effect depends on tissue composition. Fitting the absolute absorption and scattering coefficients at six wavelengths to the molar absorption spectra of the three predominant chromophores revealed that both the average total hemoglobin and oxygen saturation increased by 10%, while water concentration decreased by more than 12%. These changes indicate that the pressure-induced variation is likely due to water displacement and vascular volume increase in the region being imaged, for mild application of pressure to the breast. These results suggest that the pressure applied during optical measurements of tissue may alter the tissue physiology, and care should be taken to factor this effect into the design of optical medical instrumentation. In addition, the technique provides a unique approach to measuring tissue elastic changes in vivo in the female breast and may offer a new method for dynamic contrast imaging based on elasto-optical measurements.     

650nm光在Intralipid-10%中分布的测量和分析

本文提出一种用光纤测定光在生物组织模型Iitralipid内部光分布的方法并测定了650nm激光在Iitralipid 10%中的分布,该方法与各向同性球形光纤探头探测组织内光分布是一种互补的方法。实验结果揭示了在生物组织模型中不同方向散射光的分布规律,表明了不同的组织光学参数和不同直径的入射光束与光分布之间的关系;采用外推边界条件的漫射方程解拟合前向散射光的测量数据确定了组织模型的散射系数和吸收系数并得到较好的结果,所测量的光学参数与公开发表的结果比较其差别在2 35%以内。     

热作用致良性前列腺增生组织的光学特性变化

研究了热作用下的良性前列腺增生(BPH)组织对808和980 nm的半导体激光的光学特性的变化及其差异。实验采用双积分球测量系统以及反向倍增法获取良性前列腺增生组织的光学特性。结果表明,热作用下的良性前列腺增生组织对808 nm和980 nm的吸收系数、约化散射系数和光学穿透深度都是随着加热温度的变化而变化的,在20~80℃的温度范围内,良性前列腺增生组织对808 nm的吸收系数和约化散射系数都分别明显地较其对980 nm的吸收系数和约化散射系数要大,而其对808 nm的光学穿透深度却明显地较其对980 nm的光学穿透深度要小,吸收系数的最大值都在20℃,分别为0.528 mm-1和0.448 mm-1;最小值分别在50℃和70℃,分别为0.436 mm-1和0.326 mm-1,吸收系数的最大差异在70℃,其值为34.1%;约化散射系数的最大值都在80℃,其值分别为1.45 mm-1和1.43 mm-1,最小值分别在20℃和70℃,分别为1.15 mm-1和0.973 mm-1,最大差异在70℃,其值为24.4%;光学穿透深度的最大值分别在50℃和70℃,其值分别为0.684 mm和0.887 mm,最小值都在80℃,其值分别为0.608 mm和0.696 mm,最大差异在70℃,其值为30.4%。在70℃的热作用下良性前列腺增生组织达到完全热凝固,吸收系数、散射系数和光学穿透深度的差异达到最大值。     

Monte Carlo simulation of photon migration path in turbid media

A new method of Monte Carlo simulation is developed to simulate the photon migration path in a scattering medium after an ultrashort-pulse laser beam comes into the medium.The most probable trajectory of photons at an instant can be obtained with this method.How the photon migration paths are affected by the optical parameters of the scattering medium is analyzed.It is also concluded that the absorption coefficient has no effect on the most probable trajectory of photons.     

基于漫反射光谱的组织光学参数测量系统与方法研究

在体组织光学参数测量是生物医学光子学研究重点,不仅为人体成分无创检测、光学成像、光动力疗法等研究提供基础,并且可以快速获取人体光学参数变化,为临床诊断提供依据。研究了利用单一源探距离漫反射光谱在体测量光学参数的测量系统与反构方法。漫反射光谱测量系统由宽谱光源、高分辨光纤光谱仪及光纤探头组成,结构简单,测量方便,可准确快速测量样品漫反射光谱。在光纤探头几何形状基础上,研究了光纤收集及系统传递函数,在此基础上对反构算法进行了校正。光学参数反构算法中正向模型基于Monte Carlo以及神经网络方法,适用光学参数范围大,计算速度快;逆向算法采用主成分分析与非线性建模拟合相结合的方法,可抑制测量噪声影响。在测量系统及反构算法基础上,进行了组织仿体光学参数测量实验,结果表明,利用单一源探距离下漫反射谱,可以较为准确获取吸收系数以及约化散射系数,均方根误差分别达到4.58%以及7.92%。为保证系统测量准确性,测量波长范围应覆盖样品中所含吸收物质吸收峰范围。所研究的在体组织光学参数测量方法为人体成分无创检测及测量条件变化获取提供了基础。     

Quantitative spectroscopy of superficial turbid media

We report a novel diffuse optical spectroscopy probe design for determining optical properties of superficial volumes of turbid samples. The fiber-based probe employs a highly scattering layer placed in contact with the sample of interest. This layer diffuses photons from a collimated light source before they enter the sample and provides a basis for describing light transported in superficial media by the diffusion approximation. We compare the performance of this modified two-layer diffusion model with Monte Carlo simulations. A set of experiments that demonstrate the feasibility of this method in turbid tissue phantoms is also presented. Optical properties deduced by this approach are in good agreement with those derived by use of a benchmark method for determining optical properties. The average interrogation depth of the probe design investigated here is estimated to be less than 1 mm.     

Diffuse reflectance spectroscopy with a self-calibrating fiber optic probe

Calibration of the diffuse reflectance spectrum for instrument response and time-dependent fluctuation as well as interdevice variations is complicated, time consuming, and potentially inaccurate. We describe a novel fiber optic probe with a real-time self-calibration capability that can be used for tissue optical spectroscopy. The probe was tested in a number of liquid phantoms over a relevant range of tissue optical properties. Absorption and scattering coefficients are extracted with an average absolute error and standard deviation of 6.9%+/-7.2% and 3.5%+/-1.5%, respectively.     

Effect of absorption of multiply scattering media on the degree of residual polarization of backscattered light

The effect of absorption in a scattering medium on the degree of residual polarization of backscattered radiation is studied in the case of probing of multiply scattering media by a linearly polarized light. An approximate expression describing the dependence of the degree of residual linear polarization of the backscattered radiation on the optical characteristics of a multiply scattering medium is derived within the framework of the phenomenological approach, based on the concept of the distribution of the optical paths of partial components of the scattered optical field under the conditions of multiple scattering, and with the use of the ideas about the similarity of statistical moments of the multiply scattered optical fields. The cut-off of the partial components, characterized by a large value of the optical path, because of their absorption, results in a substantial increase of the degree of residual polarization for the bands of the selective absorption caused by the presence of chromophores in the scattering medium. The results of experiments with model scattering media (whole milk) and biological tissues (human skin in vivo) are presented.     

Photon path length distributions for an isotropically scattering planar medium

The optical path length concept is employed to determine path length distributions by an approximate numerical inversion of the Laplace transform. Six-term or 13-term closed-form representations of the hemispherical reflectance and transmittance optical path length distributions are presented. The mean path lengths and the distributions integrated over path length are found to be in excellent agreement with known results for a scattering optical depth range of 0.001 to 50.0. Any absorption feature of a medium is easily included in a scattering calculation using the results presented. Internal directional heat flux optical path length distributions are shown to exhibit the effects of higher orders of scattering.     

Correction method for influence of tissue scattering for sidestream dark-field oximetry using multicolor LEDs

We have previously proposed an estimation method of intravascular oxygen saturation \(({\rm SO}_2)\) from the images obtained by sidestream dark-field (SDF) imaging (we call it SDF oximetry) and we investigated its fundamental characteristics by Monte Carlo simulation. In this paper, we propose a correction method for scattering by the tissue and performed experiments with turbid phantoms as well as Monte Carlo simulation experiments to investigate the influence of the tissue scattering in the SDF imaging. In the estimation method, we used modified extinction coefficients of hemoglobin called average extinction coefficients (AECs) to correct the influence from the bandwidth of the illumination sources, the imaging camera characteristics, and the tissue scattering. We estimate the scattering coefficient of the tissue from the maximum slope of pixel value profile along a line perpendicular to the blood vessel running direction in an SDF image and correct AECs using the scattering coefficient. To evaluate the proposed method, we developed a trial SDF probe to obtain three-band images by switching multicolor light-emitting diodes and obtained the image of turbid phantoms comprised of agar powder, fat emulsion, and bovine blood-filled glass tubes. As a result, we found that the increase of scattering by the phantom body brought about the decrease of the AECs. The experimental results showed that the use of suitable values for AECs led to more accurate \({\rm SO}_2\) estimation. We also confirmed the validity of the proposed correction method to improve the accuracy of the \({\rm SO}_2\) estimation.     

粒状磷光增感屏点扩展函数的蒙特卡罗研究

磷光增感屏的内禀性质对磷光增感屏的成像性能影响很大.基于米氏(Mie)散射理论,计算得到磷光增感屏在不同颗粒尺寸和不同颗粒堆积密度情况下的散射系数和吸收系数.然后利用Monte Carlo(MC)方法模拟了可见光光子在磷光增感屏Gd₂O₂S:Tb(GOS)内的散射和吸收事件,得到磷光增感屏内不同位置处的可见光收集效率.利用MCNP程序模拟X射线束在磷光增感屏内的能量沉积分布,得到了金属-磷光增感屏总的点扩展函数(PSF).结果表明,在兆伏X射线成像系统中,使用小颗粒尺寸和高堆积密度的GOS磷光屏,可以改善增感屏系统的空间分辨率.     

Fourier-domain methodology for depth-selective photothermoacoustic imaging of tissue chromophores

Development of a novel photothermoacoustic (PTA) imaging technique utilizing a frequency-modulated (chirped) optical excitation and Fourier-domain methodology for depth-selective imaging of tissue chromophores is presented. Use of frequency-domain signal detection rather than short-pulse time-resolved measurements of pressure transients give an advantage of higher SNR typical to coherent detection techniques. Additionally, we introduce chirped optical excitation to generate linear frequency modulated PTA response which enables unambiguous and precise depth measurements of tissue chromophores. In order to obtain depth profilometric information from the frequency-domain PTA (FD-PTA) measurements, we describe implementation of two signal processing algorithms: matched filter compression and heterodyne mixing with coherent detection. We show that direct relationship between chromophore depth and spectrum of photothermo acoustic signals can be established to enable depth-selective tissue imaging. Application of amplitude and phase FD-PTA imaging is demonstrated in experiments with light-scattering phantoms and chicken breast tissues containing subsurface inclusions. The potential of the FD-PTA method for noninvasive tissue tomography and molecular imaging is discussed.     

Estimation of the spectral absorption of light by components of human skin

The spectra of the absorption coefficients of external radiation by basic chromophores of human skin, such as melanin, basic tissue, and blood (including oxy- and deoxyhemoglobin), are studied in the wavelength range of 300–1000 nm. For estimates, analytical methods of the theory of light transfer are used, which take into account the multilayer structure of a biological tissue, multiple light scattering in a medium, and multiple rereflections of radiation between layers. The calculated spectra are compared with the wavelength dependences of the absorption indices of these components available from the literature. It is shown that the spectral behaviors of the coefficients and indices of absorption strongly differ, which is related to the selectivity of the optical properties of a biological tissue. The possibilities of predicting the absorption coefficients of the skin components from the absorption coefficient of the entire skin measured under conditions of variation of its biophysical parameters (the volume concentrations of melanin and blood vessels) over a wide range are evaluated.     

热凝固致良性前列腺增生组织在590～1 064 nm的吸收和散射特性的变化

研究了自然的和热凝固的良性前列腺增生(BPH)组织在590～1 064 nm光谱范围的光学特性及其差异,实验采用带积分球附件的分光光度计以及反向倍增法获取组织样品的吸收和散射特性参数。结果表明：热凝固导致BPH组织在590～1 064 nm光谱范围的吸收系数明显地减小的,自然的和热凝固的BPH组织的吸收系数都有一个峰值在990 nm处,其值分别为0.438和0.416 mm-1,自然的和热凝固的BPH组织的吸收系数的最大差异在1 064 nm,其值为86.79%,其最小差异在920 nm,其值为4.74%。热凝固导致BPH组织在600～1 064 nm光谱范围的约化散射系数明显地增大,而在590 nm处,热凝固导致BPH组织的约化散射系数却是明显地减小,自然的和热凝固的BPH组织的约化散射系数都有一个峰值在970 nm处,其值分别为1.090和1.449 mm-1,其另一个峰值都在1 050 nm处,其值分别为1.116和1.627 mm-1,自然的和热凝固的BPH组织的约化散射系数的最大差异在1 060 nm,其值为47.73%,其最小差异在600 nm,其值为4.86%。     

Estimating optical properties in layered tissues by use of the Born approximation of the radiative transport equation

We use the Born approximation of the radiative transport equation to recover simultaneously the absorption and scattering coefficients in a single layer of a two-layer tissue sample from reflectance data. This method reduces the estimation of both optical properties to a single linear, least-squares problem. It is valid over length scales smaller than a transport mean free path and hence is useful for epithelial tissue layers. We demonstrate the accuracy of this method by using spatially resolved reflectance data computed with Monte Carlo simulations.     

The numerical and experimental study of photon diffusion inside biological tissue using boundary integral method

In this study, the diffusion of photons in turbid media, like biological tissue has been studied. Due to scattering and absorption of photons in such media, the study of photon propagation in biological tissue is complicated. The several numerical methods have been presented to simulate the behavior of diffused photons. Recently, Boundary Integral Method (BIM) has been offered to simulate photon migration inside biological tissues. This method has advantage, e.g. lower computational time in compared with other numerical methods. In this study, the accuracy and precision of BIM compares with another numerical method like Monte Carlo technique and finite difference method, and also the calculated results obtained by BIM and Monte Carlo method evaluate with measured results. Furthermore, the effects of scattering and absorption coefficient of tissue on the measured signal are studied.     

Influence of a skull cranial window on the measurement of haemoglobin concentration in cortical tissue by multi-spectral imaging analysis

The simplified cranial window, which is transparent thinned skull, has been used for the optical imaging of cortical tissue of small animals to measure the concentration change in haemoglobin as an intrinsic signal of brain activation. The multi-spectral images of the cortical tissue of guinea pigs through the skull cranial window were compared with those of the exposed cortex to evaluate the influence of the scattering and absorption properties of the skull on the measurement of the concentration change in haemoglobin. Although skull thickness affects the sensitivity of the optical signal due to a decrease in mean optical path length in the cortical tissue, the influence of the skull cranial window on the wavelength dependence of optical path length can be ignored when the skull thickness is less than approximately 100 mm. Accurate concentration changes in oxygenated and deoxygenated haemoglobins can be calculated from the optical signal measured through a skull cranial window and the wavelength dependence of optical path length for the exposed cortex.     

Differential optical spectroscopy for absorption characterization of scattering media

Reflectance techniques are commonly used to characterize the optical properties of tissues. However, the precise determination of local chromophore concentrations in turbid media is usually difficult because of the nonlinear dependence of light intensity as a function of scattering and absorption coefficients. A technique is presented to easily determine absorbent compound concentration ratios in a turbid media from three optical reflectance spectra, in the visible range, measured for source-detector distances less than 1cm. The validity of the method is experimentally established, in cases of sets of diluted milk containing absorbent inks, over a relatively wide range of absorption (0.05-0.5 cm(-1)) and reduced scattering (10-20 cm(-1)) coefficients.     

Absorption Coefficient Measurements of Strongly Scattering Media Using Time-Resolved Transmittance of a Short Pulse in Near-Infrared (NIR) Wavelength Range

Absorption coefficient measurements of strongly scattering and weakly absorbing media have been performed using time resolved transmittance of a 100 fs pulse through a 30 mm slab containing latex spheres suspended in water and absorbing ink solutions. The scattering and absorption coefficients were selected so that the optical properties of the media were similar to those of biological tissues. Measured curves of time-resolved transmittance of the pulse through the media were used to estimate the optical properties of the media. The experiment was made at two different wavelengths, 784 nm and 810 nm. Estimated absorption coefficients were in good agreement with those measured in a nonscattering case by a spectrophotometer.Presented at the 7th International Workshop on Multiple Scattering Lidar/Light Experiments (MUSCLE7), July 21-23 1994, Chiba, Japan.