Real-time absorption and scattering characterization of slab-shaped turbid samples obtained by a combination of angular and spatially resolved measurements

We present a fast and accurate method for real-time determination of the absorption coefficient, the scattering coefficient, and the anisotropy factor of thin turbid samples by using simple continuous-wave noncoherent light sources. The three optical properties are extracted from recordings of angularly resolved transmittance in addition to spatially resolved diffuse reflectance and transmittance. The applied multivariate calibration and prediction techniques are based on multiple polynomial regression in combination with a Newton--Raphson algorithm. The numerical test results based on Monte Carlo simulations showed mean prediction errors of approximately 0.5% for all three optical properties within ranges typical for biological media. Preliminary experimental results are also presented yielding errors of approximately 5%. Thus the presented methods show a substantial potential for simultaneous absorption and scattering characterization of turbid media.     

Measurement of optical transport properties of normal and malignant human breast tissue

We report measurement of optical transport parameters of normal and malignant (ductal carcinoma) human breast tissue. A spatially resolved steady-state diffuse reflectance technique was used for measurement of the reduced scattering coefficient (mu(s)?) and the absorption coefficient (mu(a)) of the tissue. The anisotropy parameter of scattering (g) was estimated by goniophotometric measurements of the scattering phase function. The values of mu(s)? and mu(a) for malignant breast tissue were observed to be larger than those for normal breast tissue over the wavelength region investigated (450-650 nm). Further, by using both the diffuse reflectance and the goniophotometric measurements, we estimated the Mie equivalent average radius of tissue scatterers to be larger in malignant tissue than in normal tissue.     

Determination of the surface roughness and refractive index of amorphous As40S60 films deposited by spin coating

An envelope method, based on the optical reflection spectrum taken at normal incidence, has been successfully applied to the geometrical–optical characterization of thin dielectric films having significant surface roughness. Such a method allows the determination of the average thickness and the refractive index of the films with accuracies better than 1%, as well as the average amplitude of the surface roughness with an accuracy of about 2%. Amorphous As₄₀S₆₀ thin films have been deposited by spin coating, onto glass substrates, from a solution of the bulk material in n-propylamine. Indications of the surface roughness in these films were found from total (specular plus diffuse) reflectance measurements using an integrating sphere, and also from mechanical measurements using a stylus profiler. The latter technique provided a value for the average surface roughness of 20 ± 4 nm, which is in excellent agreement with the optically determined value of 17.4 ± 0.4 nm.     

Comparison of spatially and temporally resolved diffuse-reflectance measurement systems for determination of biomedical optical properties

Time-resolved and spatially resolved measurements of the diffuse reflectance from biological tissue are two well-established techniques for extracting the reduced scattering and absorption coefficients. We have performed a comparison study of the performance of a spatially resolved and a time-resolved instrument at wavelengths 660 and 786 nm and also of an integrating-sphere setup at 550-800 nm. The first system records the diffuse reflectance from a diode laser by means of a fiber bundle probe in contact with the sample. The time-resolved system utilizes picosecond laser pulses and a single-photon-counting detection scheme. We extracted the optical properties by calibration using known standards for the spatially resolved system, by fitting to the diffusion equation for the time-resolved system, and by using an inverse Monte Carlo model for the integrating sphere. The measurements were performed on a set of solid epoxy tissue phantoms. The results showed less than 10% difference in the evaluation of the reduced scattering coefficient among the systems for the phantoms in the range 9-20 cm(-1), and absolute differences of less than 0.05 cm(-1) for the absorption coefficient in the interval 0.05-0.30 cm(-1).     

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.     

Determination of optical parameters of human breast tissue from spatially resolved fluorescence: a diffusion theory model

We report the measurement of optical transport parameters of pathologically characterized malignant tissues, normal tissues, and different types of benign tumors of the human breast in the visible wavelength region. A spatially resolved steady-state diffuse fluorescence reflectance technique was used to estimate the values for the reduced-scattering coefficient (mu(s)') and the absorption coefficient (mu(a)) of human breast tissues at three wavelengths (530, 550, and 590 nm). Different breast tissues could be well differentiated from one another, and different benign tumors could also be distinguished by their measured transport parameters. A diffusion theory model was developed to describe fluorescence light energy distribution, especially its spatial variation in a turbid and multiply scattering medium such as human tissue. The validity of the model was checked with a Monte Carlo simulation and also with different tissue phantoms prepared with polystyrene microspheres as scatterers, riboflavin as fluorophores, and methylene blue as absorbers.     

Diffuse and specular reflectance from rough surfaces

We present a reflection model for isotropic rough surfaces that have both specular and diffuse components. The surface is assumed to have a normal distribution of heights. Parameters of the model are the surface roughness given by the rms slope, the albedo, and the balance between diffuse and specular reflection. The effect of roughness on diffuse reflection is taken into account, instead of our modeling this component as a constant Lambertian term. The model includes geometrical effects such as masking and shadowing. The model is compared with experimental data obtained from goniophotometric measurements on samples of tiles and bricks. The model fits well to samples with very different reflection properties. Measurements of the sample profiles performed with a laser profilometer to determine the rms slope show that the assumed surface model is realistic. The model could therefore be used in machine vision and computer graphics to approximate reflection characteristics of surfaces. It could also be used to predict the texture of surfaces as a function of illumination and viewing angles.     

High-Transmission-Efficiency and Side-Viewing Micro OIDRS Probe for Fast and Minimally-Invasive Tumor Margin Detection

The determination of a cancer free margin I organs is a difficult and time consuming process, with an unmet need for rapid determination of tumor margin at surgery. In this paper, we report the design, fabrication and testing of a novel miniaturized optical sensor probe with "side-viewing" capability. Its unprecedented small size, unique "side-viewing" capability and high optical transmission efficiency enable the agile maneuvering and efficient data collection even in the narrow cavities inside the human body. The sensor probe consists of four micromachined substrates with optical fibers for oblique light incidence and collection of spatially resolved diffuse reflectance from the contacted tissues. The optical sensor probe has been used to conduct the oblique incidence diffuse reflectance spectroscopy (OIDRS) on a human pancreatic specimen. Based on the measurement results, the margin of the malignant tumor has been successfully determined optically, which matches well with the histological results.     

Clinical determination of tissue optical properties by endoscopic spatially resolved reflectometry

A noninvasive method to measure the optical properties of a diffusing and absorbing medium is described. Based on the spatially resolved measurement of diffuse reflectance at the sample surface, this method is particularly suitable for investigating the in vivo optical properties of biological tissues endoscopically in a clinical context. The sensitivity of the measurement is discussed, and two optical probes for two different clinical applications are presented. Preliminary measurements are performed on a nonbiological medium, which illustrate the possibilities of the proposed method. Finally, we report on in vivo measurements of the optical properties of the human esophageal wall at 630 nm.     

Determination of the refractive index of single crystal bulk samples and micro-structures

We present comparative studies for the exact determination of the refractive index of single crystals using spectroscopic ellipsometry and photonic-mode-structure investigations by means of spatially resolved photoluminescence spectroscopy, especially in the near band-gap spectral range. By applying such complementary methods we can overcome the uncertainties in the determination of the bulk refractive index introduced by surface properties. The physical effects used are the electromagnetic field reflection used by spectroscopic ellipsometry at large scale planar single crystals and the whispering-gallery-mode formation by total internal reflection in confined micro-structures. We demonstrate the applicability of such studies using the example of uniaxial ZnO bulk samples and micro-wires. By assuming a surface near region with electronic properties different from the bulk material, the method presented here gives the refractive index dispersion for both types of samples in an energy range from 1 to 3.4 eV.     

A new optical coefficient of biological tissue that determines the diffuse reflectance light distribution on the tissue surface

When a light beam is incident normally upon a tissue, the spatial distribution of diffuse reflectance on the tissue surface is affected by the optical properties of the tissue. The purpose of this paper is to find the relationship between the spatial distribution of diffuse reflectance on the tissue surface and optical properties of the tissue. The diffuse reflected light distribution is found to be determined mainly by a new optical coefficient μ_as. A qualitative relationship between μ_as and the diffuse reflected light distribution was developed by Monte Carlo simulations. Measurements of the diffuse reflected light distribution on solutions of Intralipid-10% with added ink confirm the Monte Carlo simulations. The new optical coefficient μ_as was measured from several tissue examples by the qualitative relationship. The value of μ_as was useful for determining the components of the tissue.     

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.     

Surface properties of hard protective coatings studied by optical techniques

The paper describes optical study of SiC, C and NiC layers deposited on Si substrates by double beam ion sputtering (DBIS) method. The following optical methods: ellipsometry, bidirectional reflection distribution function (BRDF) and total integrated scattering (TIS) studies have been applied. The obtained results allowed us to determine the refractive indices, extinction coefficients and the roughness parameters of DBIS films. Also surface profiles of optical constants determined from scanning ellipsometric measurements have been presented. The power spectral density functions (PSD) of surface roughness for studied samples have been determined. The influence of the deposition technology on film topography has been discussed.     

Ultrasonic reflection from a stressed interface

In seeking to develop an NDE technique for verifying the serviceability of NiTi couplers for plumbing unions, it was found that the reflection coefficient for a normally incident ultrasonic wave correlated with the stress level at the NiTi-tubing interface. Current theory pertaining to interface reflection is not complete but, even at present status, there is semiquantitative accord between theoretical predictions and normalized data for reflection coefficient versus stress level. These results are at fixed frequency and with comparable surface roughness. Variation of either frequency or surface roughness further affects the reflection coefficient. Current studies are being pursued to define this effect.     

Simultaneous extraction of optical transport parameters and intrinsic fluorescence of tissue mimicking model media using a spatially resolved fluorescence technique

We present a method based on spatially resolved fluorescence measurement for the simultaneous estimation of optical transport parameters, namely, the reduced scattering coefficient (micro s'), the absorption coefficient (micro a), and the intrinsic fluorescence spectra from turbid media. The accuracy of this approach was tested by conducting studies on a series of tissue-simulating phantoms with known optical transport properties. The estimated relative error in the values for micro s' and micro a using this technique was found to be < or =10%. Furthermore, the line shape and intensity of the intrinsic fluorescence recovered by using this approach were observed to be free from the distorting effects of the wavelength-dependent absorption and scattering properties of the medium, and they were in excellent agreement with the directly measured intrinsic fluorescence spectra of the fluorophores.     

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.     

Localization of Luminescent Inhomogeneities in Turbid Media with Spatially Resolved Measurements of cw Diffuse Luminescence Emittance

We present a steady-state method for localizing a source ofluminescence (i.e., fluorescence or phosphorescence) buried in asemi-infinite turbid medium with unknown optical properties. Adiffusion theory expression describing the emittance of an isotropicpoint source is fit to spatially resolved surface measurements of thediffuse emittance from the luminescent source. The techniquereports the location of the center of a 6.0-mm-diameter, fluorophore-containing spherical bulb embedded in a liquid phantom withan accuracy of 1.0 mm or better for source depths as great as 40.0mm. Monte Carlo data are analyzed to investigate the range and thepossible sources of error in the reconstructed source depth.     

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.     

Ion polishing with the aid of a planarizing film

An ion polishing technique employing a planarizing film in conjunction with ion-beam erosion at the planarizing angle was used to remove surface irregularities associated with conventional polishing of optical surfaces. By maintaining a planar surface throughout the erosion process, nonuniform erosion resulting from faceting, redeposition, and ion reflection is eliminated. Smooth surfaces on fused quartz are obtained by erosion of a planarizing film of photoresist at an angle of 60 degrees. The method is applicable to a wide variety of materials and may be useful for removing the surface roughness limitation on the laser-induced damage threshold of optical surfaces.     

Determination of refractive indices of opaque rough surfaces

The refractive indices of optical materials are usually determined from spectrophotometric and ellipsometric measurements of specular beams. When the roughness of the interfaces increases, the energy in the specularly reflected and transmitted beams decreases and scattering becomes predominant. For strong roughness (compared to the incident wavelength) a surface does not exhibit specular reflection or transmission, making difficult the determination of the refractive index. We describe two techniques, based on scattering measurements, that one can use to determine the refractive indices of opaque inhomogeneous media.