Depth distributions of light action spectra for skin chromophores

Light action spectra over wavelengths of 300–1000 nm are calculated for components of the human cutaneous covering: melanin, basal (bloodless) tissue, and blood oxy- and deoxyhemoglobin. The transformation of the spectra with depth in biological tissue results from two factors. The first is the wavelength dependence of the absorption coefficient corresponding to a particular skin chromophore and the second is the spectral selectivity of the radiation flux in biological tissue. This factor is related to the optical properties of all chromophores. A significant change is found to take place in the spectral distribution of absorbed radiant power with increasing depth. The action spectrum of light for the molecular oxygen contained in all components of biological tissue is also studied in the 625–645 nm range. The spectra are found to change with both the volume fraction of blood vessels and the degree of oxygenation of the blood. These results are useful for analyzing processes associated with optical absorption that are possible mechanisms for the interaction of light with biological tissues: photodissociation of oxyhemoglobin and the light-oxygen effect.     

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。     

Spectrum of the effect of laser radiation on hemoglobin of the blood vessels of skin

We suggested and consider the primary mechanism of biostimulation and of the therapeutic effect of low-level laser radiation caused by a local increase in the effectiveness of oxygen transfer by oxyhemoglobin of the blood vessels of skin. The spectra of the effect of laser radiation on oxyhemoglobin and hemoglobin of the blood vessels of skin are presented which were obtained using a method of numerical simulation with allowance for the optical characteristics of skin and for the depth of penetration into it of radiation of different wavelengths. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 65, No. 6, pp. 877–860, November–December, 1998.     

离体正常乳腺组织350～850 nm波段光谱特性

采用带有积分球附件的紫外/可见/近红外分光光度计测量了离体正常乳腺组织在350～850 nm光谱范围的反射率和透射率,运用反向倍加法得到了离体正常乳腺组织在相应光谱范围的光学参数,分析了正常乳腺组织的光学穿透深度随波长的变化情况。实验结果表明：350～850 nm波段正常乳腺组织的约化散射系数μ′_s大于吸收系数μ_a。μ′_s随着波长的增加而减小,即从350 nm波长值为9.731 mm-1～850 nm波长值为1.476 mm-1。μ_a从350 nm波长值为0.798 mm-1～850 nm波长值为0.102 mm-1,410 nm波长处存在一个吸收峰,其值为0.506 mm-1。光学穿透深度随着波长的增加而增大,从350 nm波长值为0.199 mm-1～850 nm波长值为1.439 mm。基于反向倍加法计算获得乳腺组织的光学参数,采用Monte Carlo模拟得到其相应光谱范围的反射率和透射率,并与实际测量值进行比较,二者的一致性较好。实验结果为乳腺组织的光活检及其光学治疗提供重要参考。     

Spectrophotometric system for measuring the characteristics of light scattered by biological tissues and humoral media

The structure, assembly, and physical capabilities of a spectrophotometric system intended for studying biotissues and humoral fluids are described. It can be used to investigate all the characteristics of scattered light at wavelengths of 400–1000 nm that are of interest for various problems in biomedical optics. Examples of these problems include noninvasive diagnostics of the structural and biophysical parameters of human skin tissue, analysis of the hemoglobin composition, sizes and degree of aggregation of erythrocytes, and evaluating the depth of penetration of light into biotissue. Pilot experiments on measuring the characteristics of scattered light are conducted in order to select an optimum operating mode for the system, estimate its errors, and develop ways of minimizing these errors.     

Effect of storage conditions of skin samples on their optical characteristics

The optical characteristics of skin samples are experimentally studied ex vivo and in vitro at different storage conditions. The experiments are performed on a Cary-2415 spectrophotometer in the spectral range 400–700 nm. Based on the measured diffuse reflectance and total transmittance spectra, the spectra of the absorption and reduced scattering coefficients are calculated in terms of the inverse adding-doubling method. It is shown that the method of storage of samples mainly affects the reduced scattering coefficient of biotissue. Thus, upon storage of skin in an isotonic solution and in its absence, the reduced scattering coefficient increases ∼1.5 and ∼2 times, respectively, compared to the value of this parameter for the intact sample. The differences in the absorption spectra of ex vivo samples and samples stored under different conditions are the most noticeable in the absorption range of blood and are significant above 600 nm.     

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.     

5个不同波长的激光及其线偏振激光辐照人正常膀胱组织光学特性

采用双积分球系统和光辐射测量技术的基本原理以及运用生物组织的光学模型,研究了476.5, 488, 496.5, 514.5和532 nm激光及其线偏振激光辐照人正常膀胱组织的光学特性。结果表明：组织对激光及线偏振激光的总衰减系数和散射系数均随着波长的增大而减小,而且线偏振激光与非线偏振激光入射是有明显的差异。吸收系数是随着波长的增大而缓慢地减小,但有一些起伏,而与是否线偏振光入射无明显差异。平均散射余弦也是随着波长的增大而减小,而且线偏振激光与非线偏振激光入射是有明显的差异。光学穿透深度则是随着波长的增大而增大,而有一些起伏。折射率在这5个波长范围内的值在(1.37～1.44)之间。Kubelka-Munk二流模型下组织对同一波长的激光及其线偏振激光的吸收系数、散射系数、总衰减系数和有效衰减系数没有显著性差异(P＞0.05)。组织对不同波长的激光或其线偏振激光的吸收系数、散射系数、总衰减系数和有效衰减系数则有些是有明显的差异。     

Numerical and experimental study of excited light and auto-fluorescence diffusion in teeth

Excited light and corresponding intrinsic fluorescence diffusion inside teeth tissue are an essential problem for light-based carious lesion detection. Based on finite element numerical analysis of diffusion equation, the photon density distribution of both excited light and autofluorescence of 2D premolar teeth model is obtained. The dependence of excited light and autofluorescence density distribution inside the teeth model on the scattering coefficient of enamel (5–25 mm⁻¹) and dentine (100–140 mm⁻¹) is numerically simulated and analyzed. The fitted results reveal that fluorescence intensity decreases exponentially. Optical penetration depth and fluorescence relative depth declined with the increment of scattering coefficient of enamel. And the dentine had the opposite effect. Finally, the experiment of measurement of fluorescence intensity on the teeth surface is conducted and the result is compared with the numerical computation.     

Normal and pathological color characteristics of human blood components

We have determined the averaged absorption spectra in the near UV and visible regions of the electromagnetic spectrum for aqueous solutions of hemolyzed blood, plasma, and serum from 100 donors and 95 patients with different diagnoses and different severity of their conditions. From the averaged absorption spectra, we calculated the color characteristics of the hemolyzed blood, plasma, and serum from the donors and patients by the standard CIE procedure. The blood is considered as a single, indivisible light-absorbing system in studying complex biological specimens. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 73, No. 2, pp. 220–224, March–April, 2006.     

Anomalous infrared and visible light absorption and local structure of Ag–Au core/shell nanoparticles

We measured infrared and visible light absorption spectra and EXAFS for Ag–Au core/shell particles. The shell thickness and core diameter can be evaluated from the EXAFS results, which are almost consistent with those obtained using TEM. The influence of a thinner shell only slightly appeared in the visible absorption spectra, whereas the influence appeared strongly in the infrared absorption spectra. The spectra of the material in the vicinity of the particle surface appear in the infrared spectra. On the other hand, the spectra of the rather more internal material are observed in the visible spectra. It is thought that the influence of the core metal is different in the visible spectra from the infrared spectra. By considering the penetration depth, this phenomenon can be explained.     

Isosbestic points in absorption spectra of isooctane, <Emphasis Type="Italic">n</Emphasis>-heptane,toluene, and benzene binary mixtures

Experimental absorption spectra of several pure hydrocarbons such as isooctane, n-heptane, toluene, and benzene and their binary mixtures are compared in the 1090–1240 nm and 1620–1820 nm wavelength regions, in which the second and first overtones, respectively, of vibrational frequencies of molecular groups CH, CH₂, and CH₃ are located. It is shown that in all cases there exist narrow isosbestic areas for binary mixtures specific for each of them. These areas can serve as individual characteristics of binary hydrocarbon mixtures, making qualitative analysis possible without recording the entire spectrum. Optimal wavelengths for quantitative analysis of the hydrocarbon mixture composition are determined for which the dependence between the absorption coefficient and the component concentration is approximately linear.     

Imaging of subcutaneous blood vessels and flow velocity profiles by optical coherence tomography

We have applied a compact low power rapid scanning Doppler Optical Coherence Tomography system to monitor multi-dimensional velocity profiles within the complex vessels and simultaneous real-time non-invasive imaging of skin tissues morphology in vivo, in the wavelength range of 1.3–1.5 nm. Optical clearing of skin tissues has been utilized to achieve depth of OCT images up to 1.7 mm. Current approach enables applying low-power (0.4–0.5 mW) and low-noise broadband near-infrared light sources and obtaining OCT images with down to 12 μm spatial resolution. Two-dimensional time-domain OCT images of complex flow velocity profiles in blood vessel phantom and in vivo subcutaneous human skin tissues are presented. The effect of optical clearing on in vivo images is demonstrated and discussed.     

Effect of four-photon interactions on coherent population trapping in Λ-systems

The resonance fluorescence spectrum of a Λ-system excited by two resonant light fields is calculated using a Markov analysis. Analytical formulas are derived in the strong-field limit within and beyond the rotating wave approximation. It is shown that the resonance fluorescence of the system does not vanish during coherent population trapping. Its spectrum consists of two multiplets which are similar to a triplet in the resonance fluorescence spectrum of a two-level atom and lie at the electronic transition frequencies, together with two triplets located at the frequencies of four-photon processes involving the optical excitation fields. The latter are fundamental in character and impose limits on the lower bound of the dephasing rate for the Raman resonance owing to the effect of radiative decay of the dipole transitions on the dynamics of the ground state. The effect of four-photon dephasing on the absorption spectrum of a Λ-system is analyzed and found to lead to a substantial reduction in the depth of a dip in the absorption spectrum which vanishes as the laser field strength is increased. Zh. éksp. Teor. Fiz. 113, 144–167 (January 1998)     

Oxyhemoglobin photodissociation efficiency in biological tissue exposed to laser radiation

We have obtained quantitative data on the differential (with respect to depth) and the integrated oxyhemoglobin photodissociation efficiency in the dermis when the skin surface is exposed to a light beam in the wavelength range 300–650 nm. With this aim, we have used our own previously developed optical model for skin tissue and analytical procedure for calculating the characteristics of optical fields in a medium. We have estimated the number of oxygen molecules formed at different depths in the medium, and also their integrated number over the entire thickness of the dermis as a function of the irradiation wavelength. We consider models for a dermis that is homogeneous with respect to depth and a dermis that has a layered structure. We show that the spectral photodissociation efficiency has a number of maxima associated with the absorption spectrum of oxyhemoglobin and the optical properties of all the layers of skin tissue. We discuss the effect of the epidermis on these maxima.     

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%。     

Localized light absorption by hemoglobins of an erythrocyte suspension

A correction factor accounting for the differences between the absorption spectra of an erythrocyte suspension and hemolyzed blood (hemoglobin solution) has been calculated. Erythrocytes and their aggregates are simulated by “soft” cylindrical (disk-shaped) particles. The anomalous diffraction approximation is used. At equal absorbing mass, the absorption coefficient of the suspension in the blue spectral region is shown to be capable of being several times lower than that of the solution due to localization of the absorber in erythrocytes. The hemoglobin localization effect is less important for λ _em > 600 nm. Approximate analytical equations that are valid for cylindrical particles with a length-to-diameter ratio l/d > 3–4 are proposed for calculating the correction factor. More cumbersome formulas of anomalous diffraction must be used at smaller l/d values. It is shown that these formulas transform at the limit into the analytical equations. The spectra of the correction factor for cylindrical and spherical particles are compared. The simple approximation for spherical particles is demonstrated to work well for rather long aggregates when the cylinder volume-to-surface area ratio is the same as for a spherical particle. The results can be used to solve various problems of erythrocyte optics, e.g., to reproduce sizes of aggregates by measuring their spectral absorption.     

Measurement of optical penetration depth and refractive index of human tissue

Experimental techniques for measurement of optical penetration depth and refractive index of humantissue are presented, respectively. Optical penetration depth can be obtained from the measurement ofthe relative fluence-depth distribution inside the target tissue. The depth of normal and carcinomatoushuman lung tissues irradiated with the wavelengths of 406.7, 632.8 and 674.4 nm in vitro are respectivelydetermined.In addition,a novel simple method based on total internal reflection for measuring therefractive index of biotissue in vivo is developed,and the refractive indices of skin from people of differentage, sex and skin color are measured.Their refractive indices are almost same and the average is 1.533.     

IR spectrum of the closed form of indolinospironaphthooxazine

IR absorption spectra of indolinospironaphthooxazine (ISNO) are presented. The geometric structure and normal coordinates of the ISNO molecule have been calculated using a quantum mechanical DFT method. The IR spectrum of ISNO in its original spiro form has been interpreted for the first time. Four normal modes in the range 800–1200 cm^–1 with the largest amplitudes of C_spiro–O bond change are identified. The excited ISNO molecule spiro form may experience photoconversion to the merocyanine form through them.     

Potential and restriction of high resolution,high energy photoemission in 400–8,000 eV for studying strongly correlated electron systems

InSe and InSe:Er single crystals were grown by using the Bridgman–Stockbarger method. The absorption measurements were carried out for voltage U=0 and U=30 V states of InSe and InSe:Er samples in the temperature range of 10–320 K with a step of 10 K. The absorption edge shifted towards longer wavelengths and the intensity of the absorption spectra decreased under a 5.90 kV/cm electric field. The same binding energy values for InSe and InSe:Er were calculated as 22.2 and 14.2 meV at U=0 and U=30 V, respectively. The steepness parameters and Urbach energies for InSe and InSe:Er samples increased with increasing sample temperature in the range of 10–320 K. An applied electric field caused a shift and a decrease of the intensity of the absorption spectra and an increase in the Urbach energy and steepness parameters. The shift of the absorption edge can be explained on the basis of the Franz–Keldysh effect or thermal heating of the sample under the electric field. PACS 71.20.Nr