This paper presents a preliminary application of Raman spectroscopy in conjunction with the chemometric method of partial least squares to predict silicone concentrations in homogenous turbid samples. The chemometric technique is applied to Raman spectra to develop an empirical, linear model relating sample spectra to polydimethylsiloxane (silicone) concentration. This is done using a training set of samples having optical properties and known concentrations representative of those unknown samples to be predicted. Partial least squares, performed via cross-validation, was able to predict silicone concentrations in good agreement with true values. The detection limit obtained for this preliminary investigation is similar to that reported in the magnetic resonance spectroscopy literature. The data acquisition time for this Raman-based method is 200 s which compares favourably with the 17 h acquisition required for magnetic resonance spectroscopy to obtain a similar sensitivity. The combination of Raman spectroscopy and chemometrics shows promise as a tool for quantification of silicone concentrations from turbid samples.

This paper presents a phantom which simulates the optical properties of tissue. The phantom absorption coefficient, scattering coefficient, anisotropy factor, and fluorescence quantum yield can be independently varied to investigate the effects of these parameters on fluorescence excitation and emission spectra from 300 to 650 nm. Phantom fluorophores include Flavin Adenine Dinucleotide (FAD) and Rhodamine B. Absorption is controlled by adjusting phantom hemoglobin concentration. On the basis of their smoothly varying scattering coefficient and the relatively low amount of fluorescence contributed to the mixture in comparison to other available scatterers, 1.05-ttm-diameter polystyrene microspheres were selected as a scatterer. Sample inhomogeneities are simulated by preparing the phantom in a gelatin substrate. The optical properties of turbid phantoms determined with the use of indirect techniques agree well with known values as long as μ(1 - g) > μ. Data are presented from dilute, absorbing, and turbid phantoms and inhomogeneous phantoms to qualitatively illustrate the effects of optical properties and sample geometry on fluorescence spectra. The phantom provides the framework for detailed quantitative investigations of the effects of optical properties, sample size, shape, and structure, boundary conditions, and collection geometry on fluorescence spectra. © 1993 Society for Applied Spectroscopy.

Frequency-domain photon migration (FDPM) is a widely used technique for measuring the optical properties (i.e., absorption, micro(a), and reduced scattering, micro(s)', coefficients) of turbid samples. Typically, FDPM data analysis is performed with models based on a photon diffusion equation; however, analytical solutions are difficult to obtain for many realistic geometries. Here, we describe the use of models based instead on representative samples and multivariate calibration (chemometrics). FDPM data at seven wavelengths (ranging from 674 to 956 nm) and multiple modulation frequencies (ranging from 50 to 600 MHz) were gathered from turbid samples containing mixtures of three absorbing dyes. Values for micro(a) and micro(s)' were extracted from the FDPM data in different ways, first with the diffusion theory and then with the chemometric technique of partial least squares. Dye concentrations were determined from the FDPM data by three methods, first by least-squares fits to the diffusion results and then by two chemometric approaches. The accuracy of the chemometric predictions was comparable or superior for all three dyes. Our results indicate that chemometrics can recover optical properties and dye concentrations from the frequency-dependent behavior of photon density waves, without the need for diffusion-based models. Future applications to more complicated geometries, lower-scattering samples, and simpler FDPM instrumentation are discussed.

We present a method to extend rank-annihilation-factor analysis (RAFA) for the analysis of fluorescence from homogeneous turbid samples. The method is based on a fundamental relationship between the fluorescence of a dilute solution and that of a turbid solution. We have derived this relationship, known as the transfer function, for turbid materials using the two-flux Kubelka-Munk theory. The method is tested with spectroscopic data from optically thin and turbid samples of the media of a human aorta. At 450-nm excitation, agreement between the measured and predicted dilute-solution fluorescence spectra is within 5% at all emission wavelengths; at 340-nm excitation, agreement is within 20% at all wavelengths, with some residual Soret-band absorption. The simulations presented indicate that the transfer function is markedly more sensitive to absorption than to scattering properties.

We modified and combined an open-source spatial frequency domain imaging (openSFDI) build with laser speckle imaging (LSI) into a widefield rodent cortical hemodynamic imaging system. Here, we present system specifications and in-vitro phantom measurement results.

Background

Photodynamic therapy (PDT) offers the potential for enhanced treatment of nonmelanoma skin cancer (NMSC) with minimal scarring. Yet, PDT has not achieved consistent long term effectiveness to gain widespread clinical acceptance for treatment of skin cancer. Therapeutic response varies between practitioners, patients and lesions. One important contributing factor is the absence of quantitative tools to perform in vivo dosimetry. To this end, we have developed a new quantitative imaging device that can be used to investigate parameters related to optimizing dosimetry.

Methods

We present a spatial frequency domain imaging (SFDI) based device designed to: (1) determine the optical properties at the therapeutic wavelength, which can inform variations in light penetration depth and (2) measure the spatially resolved oxygen saturation of the skin cancer lesions and surrounding tissue. We have applied this system to a preliminary clinical study of nine skin cancer lesions.

Results

Optical properties vary greatly both spatially [101%, 48% for absorption and reduced scattering, respectively] and across patients [102%, 57%]. Blood volume maps determined using visible wavelengths (460, 525, and 630 nm) represent tissue volumes within ∼1 mm in tissue (1.17 ± 0.3 mm). Here the average total hemoglobin concentration is approximately three times greater in the lesion than that detected in normal tissue, reflecting increased vasculature typically associated with tumors. Data acquired at near infrared wavelengths (730 and 850 nm) reports tissue blood concentrations and oxygenations from the underlying dermal microvasculature (volumes reaching 4.36 ± 1.32 mm into tissue).

Conclusions

SFDI can be used to quantitatively characterize in vivo tissue optical properties that could be useful for better informing PDT treatment parameters. Specifically, this information provides spatially resolved insight into light delivery into tissue and local tissue oxygenation, thereby providing more quantitative and controlled dosimetry specific to the lesion. Ultimately, by optimizing the execution of PDT, this instrument has the potential to positively improve treatment outcomes.

Accurate and timely assessment of burn wound severity is a critical component of wound management and has implications related to course of treatment. While most superficial burns and full thickness burns are easily diagnosed through visual inspection, burns that fall between these extremes are challenging to classify based on clinical appearance. Because of this, appropriate burn management may be delayed, increasing the risk of scarring and infection. Here we present an investigation that employs spatial frequency domain imaging (SFDI) and laser speckle imaging (LSI) as non-invasive technologies to characterize in-vivoburn severity. We used SFDI and LSI to investigate controlled burn wounds of graded severity in a Yorkshire pig model. Burn wounds were imaged starting at one hour after the initial injury and daily at approximately 24, 48 and 72 hours post burn. Biopsies were taken on each day in order to correlate the imaging data to the extent of burn damage as indicated via histological analysis. Changes in reduced scattering coefficient and blood flow could be used to categorize burn severity as soon as one hour after the burn injury. The results of this study suggest that SFDI and LSI information have the potential to provide useful metrics for quantifying the extent and severity of burn injuries.

Port-wine stain (PWS) birthmarks are one class of benign congenital vascular malformation. Laser therapy is the most successful treatment modality of PWS. Unfortunately, this approach has limited efficacy, with only 10% of patients experiencing complete blanching of the PWS. To address this problem, several research groups have developed technologies and methods designed to study treatment outcome and improve treatment efficacy. This article reviews seven optical imaging techniques currently in use or under development to assess treatment efficacy, focusing on: reflectance spectrophotometers/tristimulus colorimeters; laser Doppler flowmetry and laser Doppler imaging; cross-polarized diffuse reflectance colour imaging system; reflectance confocal microscopy; optical coherence tomography; spatial frequency domain imaging; and laser speckle imaging.