A method of online quantitative interpretation of diffuse reflection profiles of biological tissues

We have developed a method of combined interpretation of spectral and spatial characteristics of diffuse reflection of biological tissues, which makes it possible to determine biophysical parameters of the tissue with a high accuracy in real time under conditions of their general variability. Using the Monte Carlo method, we have modeled a statistical ensemble of profiles of diffuse reflection coefficients of skin, which corresponds to a wave variation of its biophysical parameters. On its basis, we have estimated the retrieval accuracy of biophysical parameters using the developed method and investigated the stability of the method to errors of optical measurements. We have showed that it is possible to determine online the concentrations of melanin, hemoglobin, bilirubin, oxygen saturation of blood, and structural parameters of skin from measurements of its diffuse reflection in the spectral range 450–800 nm at three distances between the radiation source and detector.     

Visualizing of skin chromophore concentrations by use of RGB images

A method is proposed for visualizing simply the concentrations of melanin, oxygenated blood, and deoxygenated blood in skin tissue using digital RGB images. The total blood concentration and oxygen saturation can also be reconstructed. Monte Carlo simulation of light transport specifies a relation between the chromophore concentrations and Commission Internationale de l'Eclairage XYZ, which are compatible with the common RGB working space. Experiments with a tissuelike agar gel phantom demonstrated the possibility of the method. In vivo imaging of a human hand during forearm occlusion demonstrated the ability of the method to evaluate hemodynamics of skin tissue.     

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.     

Depth visualization of a local blood region in skin tissue by use of diffuse reflectance images

A simple method is proposed for visualizing the depth distribution of a local blood region in skin tissue by using diffuse reflectance images at two isosbestic wavelengths of hemoglobin, 420 and 585 nm. Monte Carlo simulation of light transport specifies a relation between optical densities and the depth of the region under given concentrations of melanin in the epidermis and blood in the dermis. Phantom and in vivo experiments were performed to show the usefulness of the method.     

Evaluating a Multi-Spectral Imaging System for Mapping Pigments in Human Skin

A multi-spectral imaging system is evaluated for mapping melanin density, total-hemoglobin density, and oxygen saturation in human skin. In this system, the distribution of pigments in human skin is estimated and displayed from digital video signals using three pre-computed “look-up” tables for color conversions. The accuracy of the system is analyzed based on computer simulation by changing the number of bands, quantization levels of the camera system, and dimensions of approximation for spectral reflectance. The optimal system is examined based on the results of computer simulation.     

Explanation of Human Skin Color by Multiple Linear Regression Analysis Based on the Modified Lambert-Beer Law

Quantitative analysis of human skin color is needed in the medical and cosmetic fields. Because of the strong light scattering by biological tissues, however, analysis of skin color has not yet been fully successful. Human skin color is dominated by the colors of blood and melanin which are modified by scattering. Exposure of human skin to hot water or UV-ray changes its color because the absorbance spectrum of reflection from the human skin, and the changes in the absorbance spectra of reflection reflect the absorbance spectra of blood and melanin which are the absorption spectrum distorted by scattering. By applying the modified Lambert-Beer law, the absorbance spectrum of reflection from human skin can be expressed proportional to those distorted absorbance spectra of blood and melanin. Multiple linear regression analysis is successfully used to reproduce the absorbance spectrum of reflection from human skin from the distorted absorbance spectra of blood and melanin.     

基于扩散理论的生物组织固有荧光光谱复原方法研究

组织固有荧光光谱定义为未受生物组织吸收、散射作用影响的荧光光谱,能够直接反映组织微观结构和生物化学性质信息。为了减少吸收和散射特性对组织荧光光谱的干扰,从实测的组织荧光光谱中复原更能反映组织荧光特性的组织固有荧光光谱,搭建了基于光纤探头的组织光谱测量系统,实现生物组织相同位置处的荧光光谱和漫反射光谱测量。提出运用扩散理论从实测的漫反射光谱中提取组织生理参数,包括组织中血液体积分数、血氧饱和度、黑色素含量以及波长500 nm处约化散射系数和瑞利散射在总散射中的比例,进而计算可见波段范围内的组织光学参数;然后,根据组织光学参数和实测的漫反射光谱,从实测的荧光光谱中复原得到组织固有荧光光谱。进行临床试验验证,采集受试者皮肤组织荧光光谱与组织漫反射光谱,并复原皮肤固有荧光光谱。通过复原得到的固有荧光光谱反映人体皮肤糖基化终产物积聚量,并最终用于糖尿病无创筛查。结果显示,分别使用实测的荧光光谱和复原得到的固有荧光光谱用于糖尿病筛查时,在特异性水平同为75%时,敏感性分别为69%和90%。     

A method for operative quantitative interpretation of multispectral images of biological tissues

A method for operative retrieval of spatial distributions of biophysical parameters of a biological tissue by using a multispectral image of it has been developed. The method is based on multiple regressions between linearly independent components of the diffuse reflection spectrum of the tissue and unknown parameters. Possibilities of the method are illustrated by an example of determining biophysical parameters of the skin (concentrations of melanin, hemoglobin and bilirubin, blood oxygenation, and scattering coefficient of the tissue). Examples of quantitative interpretation of the experimental data are presented.     

Absorption spectra and light penetration depth of normal and pathologically altered human skin

A three-layered skin model (stratum corneum, epidermis, and dermis) and engineering formulas for radiative transfer theory are used to study absorption spectra and light penetration depths of normal and pathologically altered skin. The formulas include small-angle and asymptotic approximations and a layer-addition method. These characteristics are calculated for wavelengths used for low-intensity laser therapy. We examined several pathologies such as vitiligo, edema, erythematosus lupus, and subcutaneous wound, for which the bulk concentrations of melanin and blood vessels or tissue structure (for subcutaneous wound) change compared with normal skin. The penetration depth spectrum is very similar to the inverted blood absorption spectrum. In other words, the depth is minimal at blood absorption maxima. The calculated absorption spectra enable the power and irradiation wavelength providing the required light effect to be selected. Relationships between the penetration depth and the diffuse reflectance coefficient of skin (unambiguously expressed through the absorption coefficient) are analyzed at different wavelengths. This makes it possible to find relationships between the light fields inside and outside the tissue. __________ Translated from Zhurnal Prikladnoi Spektroskopii Vol. 74, No. 3, pp. 387–394, May–June, 2007.     

基于温度不敏感源-探测器距离的测量方法的模拟研究

温度变化是影响近红外无创血糖测量精度的主要因素之一。为降低温度变化对近红外漫反射光谱的影响,提出了一种基于温度不敏感源-探测器距离的测量方法,即在漫反射光强对人体组织温度变化不敏感的源-探测器距离处进行光谱测量。利用Monte Carlo方法模拟了温度为30～40 ℃、葡萄糖浓度为0～300 mmol·L-1的皮肤组织在多个源-探测器距离处的漫反射光强。根据模拟结果,分析了人体皮肤组织模型中温度不敏感源-探测器距离的存在性及其受葡萄糖浓度变化的影响;比较了1 000 nm处温度恒定和温度变动时,不同源-探测器距离处漫反射光强与葡萄糖浓度的相关性;进一步地,利用六个波长(1 000,1 050,1 100,1 150,1 350和1 410 nm)下的温度不敏感源-探测器距离及其他距离处的漫反射光强,建立了葡萄糖的偏最小二乘(PLS)模型,并比较了这些模型在温度恒定和温度变动时的预测精度。结果表明,在1 000～1 440 nm范围内,人体存在温度不敏感源-探测器距离,且葡萄糖浓度变化对该距离的影响可以忽略不计;当组织温度变化时,温度不敏感源-探测器距离处的漫反射光强与葡萄糖浓度的相关性及建模效果均明显优于其他源-探测器距离,基本接近样品温度恒定时的情况。研究表明,基于温度不敏感源-探测器距离的测量方法能有效降低温度变化对漫反射光强的影响,有望提高近红外漫反射无创血糖测量的精度。     

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。     

接触压力作用下皮肤形变对于漫反射光谱测量影响研究

人体成分无创光谱检测过程中,测量条件特别是探头压力造成的组织形变在一定程度上影响光谱测量结果。利用物理模型与仿真方法,定量分析了探头接触压力作用下皮肤组织应变,及光学参数与漫反射光谱变化。首先,在多层皮肤组织物理复合模型基础上,结合生物组织固液两相组成特性,定量分析了压力作用下各层皮肤组织形变大小以及成分变化。该模型将皮肤力学模型简化为固态弹性结构及包含于其中的牛顿流体,利用有限元方法,分别模拟不同压力作用下多层皮肤组织应变大小及时间特性,并根据各层组织体积及水含量变化,定量计算其散射系数及吸收系数改变。然后,利用Monte Carlo方法模拟压力作用前后光在组织中传输过程,定量分析了探头压力对漫反射光谱中所包含人体成分检测有效信息的影响。实验结果表明,在光纤探头接触压力作用下,人体皮肤组织中真皮层厚度变小,自由水体积减小,引起散射特性及光子传输路径改变,使得漫反射光谱中所包含的有效光谱信息下降。该研究结果为优化人体漫反射光谱测量策略,提高测量准确性以及重复性提供了参考。     

Evaluation of light scattering properties and chromophore concentrations in skin tissue based on diffuse reflectance signals at isosbestic wavelengths of hemoglobin

Optical Review - We investigate a method to evaluate light-scattering properties and chromophore concentrations in human skin tissue through diffuse reflectance spectroscopy using the reflectance...     

生物组织光传播的时域特征分析

采用蒙特卡罗方法对脉冲光在组织中传播的时域特征作了研究，并把计算结果与漫射理论的解析解作了比较，文中详细分析了组织光学特性参数与组织表面时间分辨的漫反射光分布之间的关系。结果表明：散射系影响着漫反射光强到达峰值的时间，吸收系数影响着漫反射光强在峰值之后随时间下降的速度，而ｇ因子对响应曲线的前沿影响较大，在后沿阶段基本上没有影响。     

Estimation of Absorbing Components in a Local Layer Embedded in the Turbid Media on the Basis of Visible to Near- Infrared (VIS-NIR) Reflectance Spectra

A new method is studied for estimating the hemoglobin concentration and oxygenation of a local blood layer in a human skin tissue model on the basis of visible and near infrared reflectance spectra. This method uses the iterative Monte Carlo technique for a multi-layered skin tissue model. The numerical simulation was performed to investigate the effects of estimation errors for epidermis and dermis layers on the results for the local blood layer. Experiments with skin tissue phantoms were performed to verify the possibility of this method.     

Application of transcutaneous diffuse reflectance spectroscopy in the measurement of blood glucose concentration

In this paper, the propagation characteristics of near-infrared (NIR) light in the palm tissue are analyzed,and the principle and feasibility of using transcutaneous diffuse reflectance spectroscopy for non-invasive blood glucose detection are presented. An optical probe suitable for measuring the diffuse reflectance spectrum of human palm and a non-invasive blood glucose detection system using NIR spectroscopy are designed. Based on this system, oral glucose tolerance tests are performed to measure the blood glucose concentrations of two young healthy volunteers. The partial least square calibration model is then constructed by all individual experimental data. The final result shows that correlation coefficients of the two experiments between the predicted blood glucose concentrations and the reference blood glucose concentrations are 0.9870 and 0.9854, respectively. The root mean square errors of prediction of full cross validation are 0.54 and 0.52 mmol/1, respectively.     

利用近红外光谱检测多层组织血氧饱和度的研究

利用近红外光谱无创检测生物组织血氧饱和状态,是一种极富研究和应用前景的检测技术,在临床检测中被广泛应用.但常规临床检测应用于指端仅反映局部血氧饱和度信息,在使用中具有局限性,信号的可信度也存在质疑.该文提出了一种采用反射式脉搏血氧饱和度检测技术检测生物多层组织氧合状况的新方法,该方法通过调节入射光强以适应解剖学中生物组织多层结构的检测.应用该方法针对手指结构的实验结果表明,随着入射光强的改变,反映血氧饱和状态的光电脉搏波信号有显著变化.结合手指解剖学分析表明,光电脉搏波信号的变化与手指的多层面组织结构相对应,反映不同层面血氧饱和状态.这一特点表明,通过此法可以针对生物组织的多层结构进行测量.     

Intensity of an oxygen saturation image improved using scanning NIR-LED light irradiation

A near-infrared (940 nm) image of skin tissue can be substantially enhanced using optical scanning technology (OST) that can reduce the scattering effect and remove the wrinkles or fine hair in the image. Using OST to create the skin tissue image ensured that the contour of blood vessels became more clear than that obtained from the divergent light source. In addition, the oxygen saturation of the blood vessels and tissues obtained using OST were approximately 98.65 and 93.17%, respectively, exhibiting an increase of more than 20% compared with that obtained using divergent light, and approached the value when measured using commercial pulse oximetry. Because the proposed method has a deep light penetration depth and high image intensity and resolution for oxygen saturation analysis, it is highly appropriate to be applied to future studies on diabetes and cardiovascular diseases.     

皮肤组织容积脉搏波400~1000 nm光谱特性仿真研究

根据皮肤组织解剖结构特性建立了六层层状模型,并给出了皮肤组织各层的特性参数;考虑了氧合血红蛋白和还原血红蛋白的吸收特性,依据皮肤组织各层的水、血、脂肪、血氧饱和度含量以及血管大小给出了皮肤组织各层的光谱吸收系数;对不同波长散射系数做了适当简化,给出了皮肤组织各层的光谱散射系数。利用蒙特卡罗方法仿真血管组织在收缩与舒张两种状态下, 400~1 000 nm波长光在皮肤组织多层模型中的传输过程,并通过统计大量光子的分布特性,获得了皮肤组织光谱反射系数,并利用模拟所得的两种状态下的反射系数计算得到了光谱容积脉搏波幅度。仿真结果表明,当入射光强一定时,绿光的容积脉搏波幅度优于红光和蓝光。通过计算不同波长光沿皮肤组织深度方向光能流率衰减为1/e时对应的皮肤组织深度,获得了皮肤组织光谱穿透深度。结果显示,血管舒张状态下蓝光和绿光的穿透深度较小,蓝光大部分只能达到表皮层,绿光能到达微循环层,红光可直达真皮层。考虑到光在皮肤组织中传播包含了一个从收缩到舒张的动态过程,基于此,根据穿透深度定义了脉搏波信号产生深度,利用血管舒张与收缩两种不同状态下的穿透深度计算得到了光谱产生深度。结果表明,不同波长光产生深度大于其穿透深度,蓝光产生深度较浅,且其受到的血液吸收调制较小,因而其获得的脉搏信号易受噪声干扰;红光的容积脉搏波产生深度较大,但是相比于绿光其受血液吸收调制较小,且绿光产生深度足够达到真皮血管层,因而红光容积脉搏波的幅度小于绿光。上述仿真结果明确了皮肤组织部分光谱特性,为皮肤组织多光谱容积脉搏波的精确获取及其他相关研究提供了一定的理论基础。     

漫反射光谱用于光动力治疗中鲜红斑痣皮肤氧含量监测的初步研究

尝试利用漫反射光谱检测鲜红斑痣皮肤的氧含量,并用于监测光动力治疗中鲜红斑痣皮肤氧含量的变化。采用Ocean Optics公司生产的USB2000微型光纤光谱仪采集PWS皮肤的漫反射光谱,根据氧合血红蛋白与还原血红蛋白(皮肤中两种主要的吸光基团)吸收光谱的差异,分析病变皮肤氧含量及治疗中氧含量的变化趋势。对14例PWS患者进行治疗中皮肤漫反射光谱的监测。结果显示,治疗前不同类型PWS皮肤氧含量不同,光动力治疗中大部分P4、P5型PWS皮肤氧含量没有显著降低,P6型PWS皮肤氧含量降低明显。表明现有的漫反射光谱法可定性监测PWS皮肤氧含量的变化趋势。通过进一步提高仪器的灵敏度并建立相应的光谱分析方法,有望更准确地测量PWS皮肤中的氧含量变化。