Advances in laser speckle imaging: From qualitative to quantitative hemodynamic assessment.

Laser speckle imaging (LSI) techniques have emerged as a promising method for visualizing functional blood vessels and tissue perfusion by analyzing the speckle patterns generated by coherent light interacting with living biological tissue. These patterns carry important biophysical tissue information including blood flow dynamics. The noninvasive, label-free, and wide-field attributes along with relatively simple instrumental schematics make it an appealing imaging modality in preclinical and clinical applications. The review outlines the fundamentals of speckle physics and the three categories of LSI techniques based on their degree of quantification: qualitative, semi-quantitative and quantitative. Qualitative LSI produces microvascular maps by capturing speckle contrast variations between blood vessels containing moving red blood cells and the surrounding static tissue. Semi-quantitative techniques provide a more accurate analysis of blood flow dynamics by accounting for the effect of static scattering on spatiotemporal parameters. Quantitative LSI such as optical speckle image velocimetry provides quantitative flow velocity measurements, which is inspired by the particle image velocimetry in fluid mechanics. Additionally, discussions regarding the prospects of future innovations in LSI techniques for optimizing the vascular flow quantification with associated clinical outlook are presented.

Preclinical quantitative in-vivo assessment of skin tissue vascularity in radiation-induced fibrosis with optical coherence tomography.

Radiation therapy (RT) is widely and effectively used for cancer treatment but can also cause deleterious side effects, such as a late-toxicity complication called radiation-induced fibrosis (RIF). Accurate diagnosis of RIF requires analysis of histological sections to assess extracellular matrix infiltration. This is invasive, prone to sampling limitations, and thus rarely used; instead, current practice relies on subjective clinical surrogates, including visual observation, palpation, and patient symptomatology questionnaires. This preclinical study demonstrates that functional optical coherence tomography (OCT) is a useful tool for objective noninvasive in-vivo assessment and quantification of fibrosis-associated microvascular changes in tissue. Data were collected from murine hind limbs 6 months after 40-Gy single-dose irradiation and compared with nonirradiated contralateral tissues of the same animals. OCT-derived vascular density and average vessel diameter metrics were compared to quantitative vascular analysis of stained histological slides. Results indicate that RIF manifests significant microvascular changes at this time point posttreatment. Abnormal microvascular changes visualized by OCT in this preclinical setting suggest the potential of this label-free high-resolution noninvasive functional imaging methodology for RIF diagnosis and assessment in the context of clinical RT.

Monte Carlo simulation of polarization-sensitive second-harmonic generation and propagation in biological tissue.

Polarization-sensitive second harmonic generation (p-SHG) is a nonlinear optical microscopy technique that has shown great promise in biomedicine, such as in detecting changes in the collagen ultrastructure of the tumor microenvironment. However, the complex nature of light-tissue interactions and the heterogeneity of biological samples pose challenges in creating an analytical and experimental quantification platform for tissue characterization via p-SHG. We present a Monte Carlo (MC) p-SHG simulation model based on double Stokes-Mueller polarimetry for the investigation of nonlinear light-tissue interaction. The MC model predictions are compared with experimental measurements of second-order nonlinear susceptibility component ratio and degree of polarization (DOP) in rat-tail collagen. The observed trends in the behavior of these parameters as a function of tissue thickness, as well as the overall extent of agreement between MC and experimental results, are discussed. High sensitivities of the susceptibility ratio and DOP are observed for the varying tissue thickness on the incoming fundamental light propagation pathway.

Assessment of local structural disorders of the bladder wall in partial bladder outlet obstruction using polarized light imaging.

Partial bladder outlet obstruction causes prominent morphological changes in the bladder wall, which leads to bladder dysfunction. In this paper, we demonstrate that polarized light imaging can be used to identify the location of obstruction induced structural changes that other imaging modalities fail to detect. We induced 2-week and 6-week partial outlet obstruction in rats, harvested obstructed bladders, then measured their retardances while distended to high pressures and compared them to controls. Our results show that the retardance of the central part of the ventral side (above the ureters) closer to the urethra can be used as a potential metric of the distending bladder obstruction.

Comparative study of differential matrix and extended polar decomposition formalisms for polarimetric characterization of complex tissue-like turbid media.

Development of methodologies for quantification/unique interpretation of the intrinsic polarimetry characteristics of biological tissues are important for various applications involving tissue characterization/diagnosis. A detailed comparative evaluation of the polar decomposition and the differential matrix decomposition of Mueller matrices for extraction/quantification of the intrinsic polarimetry characteristics (with special emphasis on linear retardance δ, optical rotation Ψ and depolarization Δ parameters was performed, because these are the most prominent tissue polarimetry effects) from complex tissue-like turbid media exhibiting simultaneous scattering and polarization effects. The results suggest that for media exhibiting simultaneous linear retardance and optical rotation polarization events, the use of retarder polar decomposition with its associated analysis which assumes sequential occurrence of these effects, results in systematic underestimation of δ and overestimation of Ψ parameters. Analytical relationships between the polarization parameters (δ, Ψ) extracted from both the retarder polar decomposition and the differential matrix decomposition for either simultaneous or sequential occurrence of the linear retardance and optical rotation effects were derived. The self-consistency of both decompositions is validated on experimental Mueller matrices recorded from tissue-simulating phantoms (whose polarization properties are controlled, known a-priori, and exhibited simultaneously) of increasing biological complexity. Additional theoretical validation tests were performed on Monte Carlo-generated Mueller matrices from analogous turbid media exhibiting simultaneous depolarization (Δ), linear retardance (δ) and optical rotation (Ψ) effects. After successful evaluation, the potential advantage of the differential matrix decomposition over the polar decomposition formalism was explored for monitoring of myocardial tissue regeneration following stem cell therapy.

Depolarization of light in turbid media: a scattering event resolved Monte Carlo study.

Details of light depolarization in turbid media were investigated using polarization-sensitive Monte Carlo simulations. The surviving linear and circular polarization fractions of photons undergoing a particular number of scattering events were studied for different optical properties of the turbid media. It was found that the threshold number of photon scattering interactions that fully randomize the incident polarization (defined here as <1% surviving polarization fraction) is not a constant, but varies with the photon detection angle. Larger detection angles, close to backscattering direction, show lower full depolarization threshold number for a given set of sample's optical properties. The Monte Carlo simulations also confirm that depolarization is not only controlled by the number of scattering events and detection geometry, but is also strongly influenced by other factors such as anisotropy g, medium linear birefringence, and the polarization state of the incident light.

Mueller matrix decomposition for polarized light assessment of biological tissues.

The Mueller matrix represents the transfer function of an optical system in its interactions with polarized light and its elements relate to specific biologically or clinically relevant properties. However, when many optical polarization effects occur simultaneously, the resulting matrix elements represent several "lumped" effects, thus hindering their unique interpretation. Currently, no methods exist to extract these individual properties in turbid media. Here, we present a novel application of a Mueller matrix decomposition methodology that achieves this objective. The methodology is validated theoretically via a novel polarized-light propagation model, and experimentally in tissue simulating phantoms. The potential of the approach is explored for two specific biomedical applications: monitoring of changes in myocardial tissues following regenerative stem cell therapy, through birefringence-induced retardation of the light's linear and circular polarizations, and non-invasive blood glucose measurements through chirality-induced rotation of the light's linear polarization. Results demonstrate potential for both applications.

Interstitial Doppler optical coherence tomography as a local tumor necrosis predictor in photodynamic therapy of prostatic carcinoma: an in vivo study.

We have tested the feasibility of real-time localized blood flow measurements, obtained with interstitial (IS) Doppler optical coherence tomography (DOCT), to predict photodynamic therapy (PDT)-induced tumor necrosis deep within solid Dunning rat prostate tumors. IS-DOCT was used to quantify the PDT-induced microvascular shutdown rate in s.c. Dunning prostate tumors (n=28). Photofrin (12.5 mg/kg) was administered 20 to 24 hours before tumor irradiation, with 635 nm surface irradiance of 8 to 133 mWcm(-2) for 25 minutes. High frequency ultrasound and calipers were used to measure the thickness of the skin covering the tumor and the location of the echogenic IS probe within it. A two-layer Monte Carlo model was used to calculate subsurface fluence rates within the IS-DOCT region of interest (ROI). Treatment efficacy was estimated by percent tumor necrosis within the ROI, as quantified by H&E staining, and correlated to the measured microvascular shutdown rate during PDT treatment. IS-DOCT measured significant PDT-induced vascular shutdown within the ROI in all tumors. A strong relationship (R2=0.723) exists between the percent tumor necrosis at 24 hours posttreatment and the vascular shutdown rate: slower shutdown corresponded to higher treatment efficacy, i.e., more necrosis. Controls (needle+light, no drug, n=3) showed minimal microvascular changes or necrosis (4%+/-1%). This study has correlated a biological end point with a direct and localized measurement of PDT-induced microvascular changes, suggesting a potential clinical role of on-line, real-time microvascular monitoring for optimizing treatment efficacy in individual patients.

Combined optical intensity and polarization methodology for analyte concentration determination in simulated optically clear and turbid biological media.

The use of a combined spectral intensity and polarization signals optically scattered by tissue to determine analyte concentration in optically clear and turbid biological media was explored in a simulation study. Blood plasma was chosen as the biological model and glucose as the analyte of interest. The absorption spectrum and optical rotatory dispersion were modeled using experimental data and the Drude's equation, respectively, between 500 and 2000 nm. A polarization-sensitive Monte Carlo light-propagation model was used to simulate scattering media. Unfold partial least squares and multiblock partial least squares were used as regression methods to combine the spectral intensity and polarization signals, and to predict glucose concentrations in both clear and scattering models. The results show that the combined approaches produce better predictive results in both clear and scattering media than conventional partial least squares analysis, which uses intensity or polarization spectra independently. This improvement was somewhat diminished with the addition of scattering to the model, since the polarization signals were reduced due to multiple scattering. These findings demonstrate promise for the combined approach in clear or moderately scattering biological media; however, the method's applicability to highly scattering tissues is yet to be determined. The methodology also requires experimental validation.

Stokes polarimetry in multiply scattering chiral media: effects of experimental geometry.

The spatial distribution of optical rotation alpha and surviving linear polarization fraction beta(L) of light scattered from cylindrical turbid chiral (glucose-containing) and achiral samples is studied using a linear Stokes polarimeter. alpha and beta(L) are measured in and off the incident plane as the detection angle changes from the forward to the backward direction. The experimental results exhibit a complex dependence on the detection geometry: alpha is more sensitive to glucose presence off the incident plane, whereas beta(L) exhibits larger effects in-plane, as validated by polarization sensitive Monte Carlo simulations. A rigorous methodology is presented for optimizing the experimental geometry in the polarimetric examinations of complex random systems.

Polarized light propagation in multiply scattering media exhibiting both linear birefringence and optical activity: Monte Carlo model and experimental methodology.

A Monte Carlo model for polarized light propagation in birefringent, optically active, multiply scattering media is developed in an effort to accurately represent the propagation of polarized light in biological tissue. The model employs the Jones N-matrix formalism to combine both linear birefringence and optical activity into a single effect that can be applied to photons as they propagate between scattering events. Polyacrylamide phantoms with strain-induced birefringence, sucrose-induced optical activity, and polystyrene microspheres as scattering particles are used for experimental validation. Measurements are made using a Stokes polarimeter that detects scattered light in different geometries, and compared to the results of Monte Carlo simulations run with similar parameters. The results show close agreement between the experimental measurements and Monte Carlo calculations for phantoms exhibiting turbidity and birefringence, as well as for phantoms exhibiting turbidity, birefringence, and optical activity. Other scattering-independent polarization properties can be incorporated into the developed Jones N-matrix formalism, enabling quantification of the polarization effects via an accurate polarization-sensitive Monte Carlo model.

Perturbative diffusion theory formalism for interpreting temporal light intensity changes during laser interstitial thermal therapy.

In an effort to understand dynamic optical changes during laser interstitial thermal therapy (LITT), we utilize the perturbative solution of the diffusion equation in heterogeneous media to formulate scattering weight functions for cylindrical line sources. The analysis explicitly shows how changes in detected interstitial light intensity are associated with the extent and location of the volume of thermal coagulation during treatment. Explanations for previously reported increases in optical intensity observed early during laser heating are clarified using the model and demonstrated with experimental measurements in ex vivo bovine liver tissue. This work provides an improved understanding of interstitial optical signal changes during LITT and indicates the sensitivity and potential of interstitial optical monitoring of thermal damage.

Determination of the optical properties of turbid media using relative interstitial radiance measurements: Monte Carlo study, experimental validation, and sensitivity analysis.

Interstitial quantification of the optical properties of tissue is important in biomedicine for both treatment planning of minimally invasive laser therapies and optical spectroscopic characterization of tissues, for example, prostate cancer. In a previous study, we analyzed a method first demonstrated by Dickey et al., [Phys. Med. Biol. 46, 2359 (2001)] to utilize relative interstitial steady-state radiance measurements for recovering the optical properties of turbid media. The uniqueness of point radiance measurements were demonstrated in a forward sense, and strategies were suggested for improving performance under noisy experimental conditions. In this work, we test our previous conclusions by fitting the P3 approximation for radiance to Monte Carlo predictions and experimental data in tissue-simulating phantoms. Fits are performed at: 1. a single sensor position (0.5 or 1 cm), 2. two sensor positions (0.5 and 1 cm), and 3. a single sensor position (0.5 or 1 cm) with input knowledge of the sample's effective attenuation coefficient. The results demonstrate that single sensor radiance measurements can be used to retrieve optical properties to within approximately 20%, provided the transport albedo is greater than approximately 0.9. Furthermore, compared to the single sensor fits, employing radiance data at two sensor positions did not significantly improve the accuracy of recovered optical properties. However, with knowledge of the effective attenuation coefficient of the medium, optical properties can be retrieved experimentally to within approximately 10% for an albedo greater or equal to 0.5.