Stephen J. Matcher

Quantitative assessment of skin layers absorption and skin reflectance spectra simulation in the visible and near-infrared spectral regions

We have simulated diffuse reflectance spectra of skin by assuming a wavelength-independent scattering coefficient for the different skin tissues and using the known wavelength dependence of the absorption coefficient of oxy- and deoxyhaemoglobin and water. A stochastic Monte Carlo method is used to convert the wavelength-dependent absorption coefficient and wavelength-independent scattering coefficient into reflected intensity. The absorption properties of skin tissues in the visible and near-infrared spectral regions are estimated by taking into account the spatial distribution of blood vessels, water and melanin content within distinct anatomical layers. The geometrical peculiarities of skin histological structure, degree of blood oxygenation and the haematocrit index are also taken into account. We demonstrate that when the model is supplied with reasonable physical and structural parameters of skin, the results of the simulation agree reasonably well with the results of in vivo measurements of skin spectra.

In vivo measurements of the wavelength dependence of tissue-scattering coefficients between 760 and 900 nm measured with time-resolved spectroscopy.

We present in vivo values for the optical transport coefficients (mu(a), mu(s)?) of the adult human forearm, calf, and head from 760 to 900 nm measured with time-resolved spectroscopy. The accuracy of the method is tested with tissue-simulating phantoms. We obtain mu(s)?(lambda) approximately 1.1 - (5.1 x 10(-4) lambda) mm(-1) (forearm), 1.6 - (8.9 x 10(-4) lambda) mm(-1) (calf), and 1.45 - (6.5 x 10(-4) lambda) mm(-1) (head), where lambda is measured in nanometers. At 800 nm we obtain mu(a) = 0.023 +/- 0.004 mm(-1) (forearm), 0.017 +/- 0.005 mm(-1) (calf), and 0.016 +/- 0.001 mm(-1) (head). Our values differ substantially from published in vitro data. In particular, our transport coefficients for the adult head are substantially lower than previously reported values for adult human cerebral matter and pig skull cortical bone measured in vitro.

Absolute quantification of deoxyhaemoglobin concentration in tissue near infrared spectroscopy

We describe a simple technique for non-invasively determining the absolute concentration of deoxyhaemoglobin (Hb) in living tissue using near infrared spectroscopy (NIRS). The technique uses second-differential spectroscopy to determine the relative concentration of Hb to tissue water by fitting of the spectral features of these two chromophores in the 710 nm to 840 nm region of the near infrared (NIR) spectrum. Since the concentration of tissue water is generally known with accuracies of a few per cent, one can then obtain the absolute concentration of Hb ([Hb]). The validity and likely accuracy of the technique is assessed by applying it to artificially generated NIRS data. Some clinical validation is presented by comparing, during an in vivo study, changes in [Hb] obtained by this method and those calculated using more conventional techniques of relative quantification. Finally, we discuss the likely clinical significance of the measurement of absolute Hb concentration.

Identification and Quantification of Intrinsic Optical Contrast for Near‐infrared Mammography

Near‐infrared spectroscopy has been used to quantify the composition of healthy female breast tissue in vivo. By collecting transillumination spectra in the wavelength range 680–1100 nm at 7–9 positions on the breasts of five female volunteers, an attempt was made to quantify the intra‐ and intersubject variability of breast composition. The dominant absorbers are water, lipids and hemoglobin. Hemoglobin concentration in the breast is substantially lower than that in the brain or muscle (less than 10 μM). The measured deoxyhemoglobin concentration can vary by up to 100% between different positions on the same breast. Water and lipid concentrations can show similar variability. Phantom and simulation studies demonstrate that this variability is not due to the effects of tissue boundaries on the measurements. The low hemoglobin concentration implies that optical breast imaging should be performed at wavelengths below about 850 nm to ensure that the image contrast comes predominantly from hemoglobin. Intrasubject variability could have implications for the ability of optical imaging to discern tumors from background contrast variations.

Computer simulation of the skin reflectance spectra

The reflectance spectra of the human skin in visible and near-infrared (NIR) spectral region have been calculated using the Monte Carlo technique, and the specular and internal reflection on the medium surface is taken into account. Skin is represented as a complex inhomogeneous multi-layered highly scattering and absorbing medium. The model takes into account variations in spatial distribution of blood, index of blood oxygen saturation, volume fraction of water and chromophores content. The simulation of the skin tissues optical properties and skin reflectance spectra are discussed. Comparison of the results of simulation and in vivo experimental results are given.

The collagen structure of bovine intervertebral disc studied using polarization-sensitive optical coherence tomography

Polarization-sensitive optical coherence tomography (PS-OCT) is used to measure the birefringence properties of bovine intervertebral disc and equine flexor tendon. For equine tendon the birefringence delta n is (6.0 +/- 0.2) x 10(-3) at a wavelength of 1.3 microm. This is somewhat larger than the values reported for bovine tendon. The surface region of the annulus fibrosus of a freshly excised intact bovine intervertebral disc displays an identical value of birefringence, delta n = (6.0 +/- 0.6) x 10(-3) at 1.3 microm. The nucleus pulposus does not display birefringence, the measured apparent value of delta n = (0.39 +/- 0.01) x 10(-3) being indistinguishable from the effects of depolarization due to multiple scattering. A clear difference is found between the depth-resolved retardance of equine tendon and that of bovine intervertebral disc. This apparently relates to the lamellar structure of the latter tissue, in which the collagen fibre orientation alternates between successive lamellae. A semi-empirical model based on Jones calculus shows that the measurements are in reasonable agreement with previous optical and x-ray data. These results imply that PS-OCT could be a useful tool to study collagen organization within the intervertebral disc in vitro and possibly in vivo and its variation with applied load and disease.

Modelling the sampling volume for skin blood oxygenation measurements

The absolute quantified measurement of haemoglobin skin blood saturation from collected reflectance spectra of the skin is complicated by the fact that the blood content of tissues can vary both in the spatial distribution and in the amount. These measurements require an understanding of which vascular bed is primarily responsible for the detected signal. Knowing the spatial detector depth sensitivity makes it possible to find the best range of different probe geometries for the measurements of signal from the required zones and group of vessels inside the skin. To facilitate this, a Monte Carlo simulation has been developed to estimate the sampling volume offered by fibre-optic probes with a small source-detector spacing (in the current report 250 μm, 400 μm and 800 μm). The optical properties of the modelled medium are taken to be the optical properties of the Caucasian type of skin tissue in the visible range of the spectrum. It is shown that, for a small source-detector separation (800 μm and smaller), rough boundaries between layers of different refractive index can play a significant role in skin optics. Wavy layer interfaces produce a deeper and more homogeneous distribution of photons within the skin and tend to suppress the direct channelling of photons from source to detector. The model predicts that a probe spacing of 250 μm samples primarily epidermal layers and papillary dermis, whereas spacings of 400–800 μm sample upper blood net dermis and dermis.

Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy

Second harmonic generation (SHG) and two-photon fluorescence (TPF) microscopy is used to image the intercellular and pericellular matrix in normal and degenerate equine articular cartilage. The polarization sensitivity of SHG can be used directly to determine fiber orientation in the superficial 10 to 20 microm of tissue, and images of the ratio of intensities taken with two orthogonal polarization states reveal small scale variations in the collagen fiber organization that have not previously been reported. The signal from greater depths is influenced by the birefringence and biattenuance of the overlying tissue. An assessment of these effects is developed, based on the analysis of changes in TPF polarization with depth, and the approach is validated in tendon where composition is independent of depth. The analysis places an upper bound on the biattenuance of tendon of 2.65 x 10(-4). Normal cartilage reveals a consistent pattern of variation in fibril orientation with depth. In lesions, the pattern is severely disrupted and there are changes in the pericellular matrix, even at the periphery where the tissue appears microscopically normal. Quantification of polarization sensitivity changes with depth in cartilage will require detailed numerical models, but in the meantime, multiphoton microscopy provides sensitive indications of matrix changes in cartilage degeneration.

Measurement of bone mineral density via light scattering

In this study we have investigated the potential of optical techniques to monitor changes in bone mineral density (BMD) via changes in scattering coefficient. For each of five bone samples, diffuse reflection and transmission coefficients were measured over the wavelength range 520-960 nm using an integrating sphere and CCD spectrometer. These were converted into optical absorption and scattering coefficients using a Monte Carlo inversion procedure. Measurements were made on samples immersed in formic acid solution for different lengths of time in order to investigate the effect of reduction in BMD on the optical properties. After full demineralization, the optical scattering coefficient fell by a factor 4. From the observed degree of fluctuation of the measurements, we estimate that BMD could be measured with an accuracy of 7% if optical scattering can be measured with an accuracy of 10%. We also report preliminary measurements of bone scattering using optical coherence tomography (OCT). An inter-side variability of 3% is obtained on dry samples with and without overlying periosteum. These results suggest that minimally invasive techniques for measuring optical scattering, such as OCT, may have a role in monitoring regional changes in BMD. This could be an important advance in our understanding of bone remodelling and its relationship to osteoarthritis. Both the integrating sphere and OCT measurements also suggest that light transport in bone is spatially anisotropic. OCT was used to assess probability of obtaining results in vivo.

Polypyrrole Nanoparticles: A Potential Optical Coherence Tomography Contrast Agent for Cancer Imaging

A near-infrared (NIR) absorbing contrast agent based on polypyrrole nanoparticles is described. Quantitative optical coherence tomography studies on tissue phantoms and Mie scattering calculations indicate their potential application for early-stage cancer diagnosis.