Determination of optical coefficients of biological tissue from a single integrating-sphere

The detection of interactions between light and tissue can be used to characterize the optical properties of the tissue. The development is described of a method that determines optical coefficients of biological tissue from a single optical reflectance spectrum measured with an integrating-sphere. The experimental system incorporated a DH-2000 deuterium tungsten halogen light source, a USB4000-VIS-NIR miniature fiber optic spectrometer and an integrating-sphere. Fat emulsion and ink were used to mimic the scattering and absorbing properties of tissue in the tested sample. The measured optical reflectance spectrums with different scattering and absorbing properties were used to train a back-propagation neural network (BPNN). Then the neural network (BPNN) was used to determine the optical coefficients of biological tissue from a single optical reflectance spectrum measured with an integrating-sphere. Tests on tissue-simulation phantoms showed the relative errors of this technique to be 7% for the reduced scattering coefficient and 15% for the absorption coefficients. The optical properties of human skin were also measured in vivo.     

Spatially resolved absolute diffuse reflectance measurements for noninvasive determination of the optical scattering and absorption coefficients of biological tissue

The absorption and transport scattering coefficients of biological tissues determine the radial dependence of the diffuse reflectance that is due to a point source. A system is described for making remote measurements of spatially resolved absolute diffuse reflectance and hence noninvasive, noncontact estimates of the tissue optical properties. The system incorporated a laser source and a CCD camera. Deflection of the incident beam into the camera allowed characterization of the source for absolute reflectance measurements. It is shown that an often used solution of the diffusion equation cannot be applied for these measurements. Instead, a neural network, trained on the results of Monte Carlo simulations, was used to estimate the absorption and scattering coefficients from the reflectance data. Tests on tissue-simulating phantoms with transport scattering coefficients between 0.5 and 2.0 mm(-1) and absorption coefficients between 0.002 and 0.1 mm(-1) showed the rms errors of this technique to be 2.6% for the transport scattering coefficient and 14% for the absorption coefficients. The optical properties of bovine muscle, adipose, and liver tissue, as well as chicken muscle (breast), were also measured ex vivo at 633 and 751 nm. For muscle tissue it was found that the Monte Carlo simulation did not agree with experimental measurements of reflectance at distances less than 2 mm from the incident beam.     

Fiber-optic probe for noninvasive real-time determination of tissue optical properties at multiple wavelengths

We present a compact, fast, and versatile fiber-optic probe system for real-time determination of tissue optical properties from spatially resolved continuous-wave diffuse reflectance measurements. The system collects one set of reflectance data from six source-detector distances at four arbitrary wavelengths with a maximum overall sampling rate of 100 Hz. Multivariate calibration techniques based on two-dimensional polynomial fitting are employed to extract and display the absorption and reduced scattering coefficients in real-time mode. The four wavelengths of the current configuration are 660, 785, 805, and 974 nm, respectively. Cross-validation tests on a 6 x 7 calibration matrix of Intralipid-dye phantoms showed that the mean prediction error at, e.g., 785 nm was 2.8% for the absorption coefficient and 1.3% for the reduced scattering coefficient. The errors are relative to the range of the optical properties of the phantoms at 785 nm, which were 0-0.3/cm for the absorption coefficient and 6-16/cm for the reduced scattering coefficient. Finally, we also present and discuss results from preliminary skin tissue measurements.     

Monte Carlo-based inverse model for calculating tissue optical properties. Part I: Theory and validation on synthetic phantoms

A flexible and fast Monte Carlo-based model of diffuse reflectance has been developed for the extraction of the absorption and scattering properties of turbid media, such as human tissues. This method is valid for a wide range of optical properties and is easily adaptable to existing probe geometries, provided a single phantom calibration measurement is made. A condensed Monte Carlo method was used to speed up the forward simulations. This model was validated by use of two sets of liquid-tissue phantoms containing Nigrosin or hemoglobin as absorbers and polystyrene spheres as scatterers. The phantoms had a wide range of absorption (0-20 cm(-1)) and reduced scattering coefficients (7-33 cm(-1)). Mie theory and a spectrophotometer were used to determine the absorption and reduced scattering coefficients of the phantoms. The diffuse reflectance spectra of the phantoms were measured over a wavelength range of 350-850 nm. It was found that optical properties could be extracted from the experimentally measured diffuse reflectance spectra with an average error of 3% or less for phantoms containing hemoglobin and 12% or less for phantoms containing Nigrosin.     

Influence of layered tissue architecture on estimates of tissue optical properties obtained from spatially resolved diffuse reflectometry

Most instruments used to measure tissue optical properties noninvasively employ data-analysis algorithms that rely on the simplifying assumption that the tissue is semi-infinite and homogeneous. The influence of a layered tissue architecture on the determination of the scattering and absorption coefficients has been investigated in this study. Reflectance as a function of distance from a point source for a two-layered tissue architecture that simulates skin overlying fat was calculated by using a Monte Carlocode. These data were analyzed by using a diffusion theory modelfor a homogeneous semi-infinite medium to calculate the scatter and absorption coefficients. Depending on the algorithm and the radial distance, the estimated tissue optical properties were different from those of either layer, and under some circumstances, physically impossible. In addition, the sensitivity and cross talk of the estimated optical properties to changes in input optical properties were calculated for different layered geometries. For typical optical properties of skin, the sensitivity to changes in optical properties is highly dependent on the layered architecture, the measurement distance, and the fitting algorithm. Furthermore, a change in the input absorption coefficient may result in an apparent change in the measured scatter coefficient, and a change in the in put scatter coefficient may result in an apparent change in the measured absorption coefficient.     

Integrating the digitized backscattered image to measure absorption and reduced-scattering coefficients in vivo

Measurement of absorption and reduced-scattering optical coefficients a and s is possible when a steady-state backscattered image is used on a sample surface. A new method for processing the backscattered image, acquired with a CCD, has been developed. The image is integrated to decrease sensitivity to noise. The resulting curve is defined as the integral reflectance. The curve is then fitted with a relaxation model to evaluate a and s . We have validated the method with calibrated scattering and absorption phantoms. The integral reflectance method is then applied to measurements of the a and s coefficients of human skin in vivo .     

In vivo determination of local skin optical properties and photon path length by use of spatially resolved diffuse reflectance with applications in laser Doppler flowmetry

Methods for local photon path length and optical properties estimation, based on measured and simulated diffuse reflectance within 2 mm from the light source, are proposed and evaluated in vivo on Caucasian human skin. The accuracy of the methods was good (2%-7%) for path length and reduced scattering but poor for absorption estimation. Reduced scattering and absorption were systematically lower in the fingertip than in the forearm skin (633 nm). A maximum intrasite and interindividual variation of approximately 35% in an average photon path length was found. The methodology was applied in laser Doppler flowmetry, where path-length normalization of the estimated perfusion removed the optical property dependency.     

Determination of the optical properties of two-layer turbid media by use of a frequency-domain hybrid monte carlo diffusion model

The general two-layer inverse problem in biomedical photon migration is to estimate the absorption and scattering coefficients of each layer as well as the top-layer thickness. We attempted to solve this problem, using experimental and simulated spatially resolved frequency-domain (FD) reflectance for optical properties typical of skin overlying muscle or skin overlying fat in the near infrared. Two forward models of light propagation were used: a two-layer diffusion solution [Appl. Opt. 37, 779 (1998)] and a hybrid Monte Carlo (MC) diffusion model [Appl. Opt. 37, 7401 (1998)]. MC-simulated FD reflectance data were fitted as relative measurements to the hybrid and the pure diffusion models. It was found that the hybrid model could determine all the optical properties of the two-layer media studied to ~5%. Also, the same accuracy could be achieved by means of fitting MC-simulated cw reflectance data as absolute measurements, but fitting them as relative ones is an ill-posed problem. In contrast, two-layer diffusion could not retrieve the top-layer optical properties as accurately for FD data and was ill-posed for both relative and absolute cw data. The hybrid and the pure diffusion models were also fitted to experimental FD reflectance measurements from two-layer tissue-simulating phantoms representative of skin-on-fat and skin-on-muscle baseline optical properties. Both the hybrid and the diffusion models could determine the optical properties of the lower layer. The hybrid model demonstrated its potential to retrieve quantitatively the transport scattering coefficient of skin (the upper layer), which was not possible with the pure diffusion model. Systematic discrepancies between model and experiment may compromise the accuracy of the deduced top-layer optical properties. Identifying and eliminating such discrepancies is critical to practical application of the method.     

Sequential estimation of optical properties of a two-layered epithelial tissue model from depth-resolved ultraviolet-visible diffuse reflectance spectra

A method for estimating the optical properties of two-layered media (such as squamous epithelial tissue) over a range of wavelengths in the ultraviolet-visible spectrum is proposed and tested with Monte Carlo modeling. The method first used a fiber-optic probe with angled illumination and the collection fibers placed at a small separation (or=1000 microm) was used to detect diffuse reflectance preferentially from the bottom layer. A second Monte Carlo-based inverse model for a two-layered medium was applied to estimate the bottom layer optical properties, as well as the top layer thickness, given that the top layer optical properties have been estimated. The results of Monte Carlo validation show that this method works well for an epithelial tissue model with a top layer thickness ranging from 200 to 500 microm. For most thicknesses within this range, the absorption coefficients were estimated to within 15% of the true values, the reduced scattering coefficients were estimated to within 20% and the top layer thicknesses were estimated to within 20%. The application of a variance reduction technique to the Monte Carlo modeling proved to be effective in improving the accuracy with which the optical properties are estimated.     

A feature parameter of optical diffuse reflectance on normal and malignant human breast tissue

Light that is delivered to the tissue surface undergoes multiple elastic scattering and absorption; part of it returns to the surface as optical diffuse reflectance, carrying quantitative information on the structure and composition of the measured tissue. In this paper, we utilized a well tested and publicly available Monte Carlo simulation software to simulate optical diffuse reflectance on normal and malignant human breast tissue in the visible wavelength region 450–650 nm. Based on the Monte Carlo simulation results, we discovered a feature parameter of optical diffuse reflectance on the simulated tissue surface. Normal and malignant human breast tissue may be discriminated by the value of the feature parameter. The values of the feature parameter are shown for normal and malignant human breast tissue in the visible wavelength region 450–650 nm. Experiments with tissue phantoms showed that the feature parameter is correlated to the component of the phantom. So the feature parameter is useful for the non-invasive optical diagnosis of biological tissue.     

Why do veins appear blue? A new look at an old question

We investigate why vessels that contain blood, which has a red or a dark red color, may look bluish in human tissue. A CCD camera was used to make images of diffusely reflected light at different wavelengths. Measurements of reflectance that are due to model blood vessels in scattering media and of human skin containing a prominent vein are presented. Monte Carlo simulations were used to calculate the spatially resolved diffuse reflectance for both situations. We show that the color of blood vessels is determined by the following factors: (i) the scattering and absorption characteristics of skin at different wavelengths, (ii) the oxygenation state of blood, which affects its absorption properties, (iii) the diameter and the depth of the vessels, and (iv) the visual perception process.     

In vivo local determination of tissue optical properties: applications to human brain

Local and superficial near-infrared (NIR) optical-property characterization of turbid biological tissues can be achieved by measurement of spatially resolved diffuse reflectance at small source-detector separations (<1.4 mm). However, in these conditions the inverse problem, i.e., calculation of localized absorption and the reduced scattering coefficients, is necessarily sensitive to the scattering phase function. This effect can be minimized if a new parameter of the phase function gamma, which depends on the first and the second moments of the phase function, is known. If gamma is unknown, an estimation of this parameter can be obtained by the measurement, but the uncertainty of the absorption coefficient is increased. A spatially resolved reflectance probe employing multiple detector fibers (0.3-1.4 mm from the source) is described. Monte Carlo simulations are used to determine gamma, the reduced scattering and absorption coefficients from reflectance data. Probe performance is assessed by measurements on phantoms, the optical properties of which were measured by other techniques [frequency domain photon migration (FDPM) and spatially resolved transmittance]. Our results show that changes in the absorption coefficient, the reduced scattering coefficient, and gamma can be measured to within +/-0.005 mm(-1), +/-0.05 mm(-1), and +/-0.2, respectively. In vivo measurements performed intraoperatively on a human skull and brain are reported for four NIR wavelengths (674, 811, 849, 956 nm) when the spatially resolved probe and FDPM are used. The spatially resolved probe shows optimum measurement sensitivity in the measurement volume immediately beneath the probe (typically 1 mm(3) in tissues), whereas FDPM typically samples larger regions of tissues. Optical-property values for human skull, white matter, scar tissue, optic nerve, and tumors are reported that show distinct absorption and scattering differences between structures and a dependence on the phase-function parameter gamma.     

Optical characterization method for black pigments applied to solar-selective absorbing paints

We propose a novel, to our knowledge, method for characterizing the optical properties of pigment particles or powders. Measurements of the diffuse and the total transmittance as well as the diffuse and the total reflectance are used to obtain effective scattering and absorption coefficients per unit length for the particles that are dispersed in a continuous matrix. For dilute dispersions in the single-scattering regime scattering and absorption cross sections of the particles were obtained. The method was applied to two pigments, namely, FeMnCuO(x) and black carbon. The data were obtained by use of pellets consisting of low concentrations of FeMnCuO(x) or black-carbon pigments dispersed in a KBr matrix. The pigment volume concentrations used to evaluate the scattering and the absorption coefficients ranged from 0.053% to 0.530% for FeMnCuO(x) and 0.076% to 0.310% for the black carbon. These ranges were found to exhibit the linear dependence of the coefficients as a function of volume fraction, as given by single-scattering theory.     

Analytical model of light reflectance for extraction of the optical properties in small volumes of turbid media

Monte Carlo simulations and experiments in tissue phantoms were used to empirically develop an analytical model that characterizes the reflectance spectrum in a turbid medium. The model extracts the optical properties (scattering and absorption coefficients) of the medium at small source-detector separations, for which the diffusion approximation is not valid. The accuracy of the model and the inversion algorithm were investigated and validated. Four fiber probe configurations were tested for which both the source and the detector fibers were tilted at a predetermined angle, with the fibers parallel to each other. This parallel-fiber geometry facilitates clinical endoscopic applications and ease of fabrication. Accurate extraction of tissue optical properties from in vivo spectral measurements could have potential applications in detecting, noninvasively and in real time, epithelial (pre)cancers.     

Optical properties of an optically rough coating from inversion of diffuse reflectance measurements

A method is developed for determining the optical properties of an optically rough coating on an opaque substrate from reflectance measurements. A modified Kubelka-Munk two- flux model is used to calculate the reflectance of the coating as a function of the refractive index, absorption coefficient, scattering coefficient, and thickness. The calculated reflectance is then fitted to measurements using a spectral projected gradient algorithm, allowing the optical properties to be obtained. The technique is applied to titanium dioxide coatings on a titanium substrate. Realistic values of refractive index and absorption coefficients are generally obtained. Quantities that are useful for solar water-splitting applications are calculated, including the depth profile of absorption and the proportion of the incident photon flux absorbed in the coating under solar illumination.     

Noninvasive measurement of fluorophore concentration in turbid media with a simple fluorescence /reflectance ratio technique

Measurement of the concentration of fluorescent compounds in turbid media is difficult because the absorption and multiple scattering of excitation and emission of light has a large effect on the detected fluorescence. For surface measurements with optical fibers we demonstrate by experiments and numerical simulation that this effect can be minimized by measurement of the fluorescence at one source-detector distance, the diffusely reflected excitation light at a second distance, and with the ratio of these two signals as an indicator of fluorophore concentration. For optical properties typical of soft tissue in the red and the near infrared the optimum performance is obtained by measurement of fluorescence at 0.65 mm and reflectance at 1.35 mm. This choice reduces the rms error in fluorophore concentration to 14.6% over a wide range of absorption and scattering coefficients.     

基于微扰蒙特卡罗的薄层状组织光学参数重构技术

针对层状组织体的光学参数（吸收系数和散射系数）快速重构问题展开讨论.发展了基于漫反射测量的适合于胃、宫颈等内脏薄层状组织光学参数重构的微扰蒙特卡罗模拟.数值模拟结果显示,该方法能够有效地利用漫反射光强重构临床应用范围内的光学参数,对单层模拟体的吸收系数和散射系数的重构误差均不大于±3%,对双层模拟体的重构误差小于10%,且两参数的重构之间几乎没有互扰.使用固体模型对上述算法进行的实验显示,重构出的散射参数与真值的误差小于±10%.进行一组光学参数重构耗时不超过1 min.结果表明,该方法可快速、准确地重构光学参数,为内脏器官近红外早期癌症诊断的临床应用提供保障.     

Measurement of the absorption and scattering properties of turbid liquid foods using hyperspectral imaging

A hyperspectral imaging system in line scanning mode was used for measuring the absorption and scattering properties of turbid food materials over the visible and near-infrared region of 530-900 nm. An instrumental calibration procedure was developed to compensate for the nonuniform instrument response of the imaging system. A nonlinear curve-fitting algorithm for a steady-state diffusion theory model was proposed to determine absorption (mua) and reduced scattering coefficients (mu's) from the spatially resolved hyperspectral reflectance profiles. The hyperspectral imaging system provided good measurement of mua and mu's for the simulation samples made of Intralipid scattering material and three absorbers (blue dye, green dye, and black ink) with average fitting errors of 16% and 11%, respectively. The optical properties of the fruit and vegetable juices and milks were determined. Values of the absorption and reduced scattering coefficient at 600 nm were highly correlated to the fat content of the milk samples with the correlation coefficient of 0.995 and 0.998, respectively. Compared to time-resolved and frequency-domain techniques, the hyperspectral imaging technique provides a faster and simpler means for measuring the optical properties of turbid food and agricultural products.     

Polarized light reflection from strained sinusoidal surfaces

We propose optical polarization imaging as a minimally invasive technique for measuring the mechanical properties of plastics and soft tissues through their change in reflectance properties with applied strain or force. We suggest that changes in surface roughness are responsible for the linear reflectivity changes with applied stretch or strain. Several aspects of this model are tested, including the dependence on the angle of incidence, the change in scattering and absorption coefficients with strain, and the lateral spatial resolution. The application of the technique to multilayer structures such as skin and competing optical effects such as laser speckle are discussed.     

Imaging with diffusing light: an experimental study of the effect of background optical properties

The overall image quality and diagnostic potential of time-resolved transmittance imaging depend on sensitivity to optical contrast, capacity to discriminate scattering from absorption contributions, and spatial resolution. We have investigated experimentally the effects of the optical properties of the background medium on the overall image quality of optical imaging based on fitting the experimental data to the solution of the diffusion equation and on time gating. Images were acquired from phantoms with different background optical properties, while the optical contrast between inhomogeneities and background is kept constant. Data were collected every 0.2 cm over a 6 cm x 6 cm area from realistic tissue phantoms containing cylindrical inhomogeneities (1 cm high and 1 cm in diameter) embedded in a 5-cm-thick turbid slab. The optical coefficients of the background were varied in the ranges of 5-15 cm(-1) for transport scattering and 0.02-0.08 cm(-1) for absorption. The optical contrast for the inclusions was kept at values of -50% and +50% for the scattering and -75% and +300% for the absorption. The results show that both high scattering and high absorption are beneficial.