This PDF file contains the front matter associated with SPIE Proceedings Volume 8952, including the Title Page, Copyright Information, Table of Contents, and the Conference Committee listing.

A novel two-frequency dynamic light scattering system (TF-DLS) and differential heterodyne en face laser Doppler velocimeter (LDV) are proposed and setup. The power spectrum of the heterodyne beat signal is detected whereas the width of power spectrum and shift of central frequency are measured simultaneously. These are able to provide the particle sizing and its tangential velocity in suspension. Finally, the localization ability of TF-DLS/LDV system on en face velocity measurement was discussed.

Experimental data on detection and identification of variety of biochemical agents, such as proteins (albumin, interferon, C reactive protein), microelements (Na+, Ca+), antibiotic of different generations, in both single and multi component solutions under varied in wide range concentration are represented. Analysis has been performed on the light scattering parameters of whispering gallery mode (WGM) optical resonance based sensor with dielectric microspheres from glass and PMMA as sensitive elements fixed by spin - coating techniques in adhesive layer on the surface of substrate or directly on the coupling element. Sensitive layer was integrated into developed fluidic cell with a digital syringe. Light from tuneable laser strict focusing on and scattered by the single microsphere was detected by a CMOS camera. The image was filtered for noise reduction and integrated on two coordinates for evaluation of integrated energy of a measured signal. As the entrance data following signal parameters were used: relative (to a free spectral range) spectral shift of frequency of WGM optical resonance in microsphere and relative efficiency of WGM excitation obtained within a free spectral range which depended on both type and concentration of investigated agents. Multiplexing on parameters and components has been realized using spatial and spectral parameters of scattered by microsphere light with developed data processing. Biochemical component classification and identification of agents under investigation has been performed by network analysis techniques based on probabilistic network and multilayer perceptron. Developed approach is demonstrated to be applicable both for single agent and for multi component biochemical analysis.

Photopolymerized hydrogels are commonly used for a broad range of biomedical applications. As long as the polymer volume is accessible, gels can easily be hardened using light illumination. However, in clinics, especially for minimally invasive surgery, it becomes highly challenging to control photopolymerization. The ratios between polymerizationvolume and radiating-surface-area are several orders of magnitude higher than for ex-vivo settings. Also tissue scattering occurs and influences the reaction. We developed a Monte Carlo model for photopolymerization, which takes into account the solid/liquid phase changes, moving solid/liquid-boundaries and refraction on these boundaries as well as tissue scattering in arbitrarily designable tissue cavities. The model provides a tool to tailor both the light probe and the scattering/absorption properties of the photopolymer for applications such as medical implants or tissue replacements. Based on the simulations, we have previously shown that by adding scattering additives to the liquid monomer, the photopolymerized volume was considerably increased. In this study, we have used bovine intervertebral disc cavities, as a model for spinal degeneration, to study photopolymerization in-vitro. The cavity is created by enzyme digestion. Using a custom designed probe, hydrogels were injected and photopolymerized. Magnetic resonance imaging (MRI) and visual inspection tools were employed to investigate the successful photopolymerization outcomes. The results provide insights for the development of novel endoscopic light-scattering polymerization probes paving the way for a new generation of implantable hydrogels.

We have recently demonstrated a means for quantifying the absorption and scattering properties of biological tissue through multidiameter single-fiber reflectance (MDSFR) spectroscopy. These measurements can be used to correct single-fiber fluorescence (SFF) spectra for the influence of optical properties, enabling quantification of intrinsic fluorescence. In our previous work, we have used a series of pinholes to show that selective illumination and light collection using a coherent fiber bundle can simulate a single solid-core optical fiber with variable diameter for the purposes of MDSFR spectroscopy. Here, we describe the construction and validation of a clinical MDSFR/SFF spectroscopy system that avoids the limitations encountered with pinholes and free-space optics. During one measurement, the new system acquires reflectance spectra at the effective diameters of 200, 600, and 1000 μm, and a fluorescence spectrum at an effective diameter of 1000 μm. From these spectra, we measure the absolute absorption coefficient, μ_a, reduced scattering coefficient, μ’_s, phase function parameter, γ, and intrinsic fluorescence, Qμ^f_a, across the measured spectrum. We validate the system using Intralipid- and polystyrene sphere-based scattering phantoms, with and without the addition of the absorber Evans Blue. Finally, we demonstrate the combined MDSFR/SFF of phantoms with varying concentrations of Intralipid and fluorescein, wherein the scattering properties are measured by MDSFR and used to correct the SFF spectrum for accurate quantification of Qμ^f_a.

In order to increase sensitivity in the depth of diffusive media and to separate chromophores with distinct spectral signatures, we developed a method to process time-domain/multi-wavelength diffuse optical acquisitions: 3D Reconstructions of chromophore concentrations are performed with an algorithm based on the use of Mellin-Laplace Transform and material basis. A noise weighted data matching term is optimized by using the conjugated gradients method without expressing the Jacobian matrix of the system. As the algorithm uses reference measurements on a known medium, it does not require measurements or computations of the instrument response function of the system. Validations are performed in the reflectance geometry on a tissue-mimicking phantom composed of intralipid and black ink and a cylindrical blue dye inclusion with a radius of 4mm located at 15mm in depth. The optical tomography setup includes a laser whose picosecond pulses are injected via an optical fiber to the probed diffusive medium and the light collected by two fibers (located 15mm apart from the source), is sent to a Single-Photon Avalanche Diode (SPAD) connected to a Time-Correlated Single-Photon Counting (TCSPC) board. The source and two detectors scan the surface of the medium so as to provide 30 source-detector couples, 900 time-bins and 5 wavelength signals. 3D reconstructions performed on the black ink and blue dye materials on a mesh of around 10000 nodes show that we are able to detect, localize and determine the composition of the inclusion and the background.

Modeling the propagation of coherent polarized light through a turbid scattering medium using the Monte Carlo method enables better understanding of the peculiarities of image/signal formation in modern optical diagnostic techniques, such as optical coherence tomography (OCT), coherent/enhanced backscattering, laser speckle imaging and diffusing-wave spectroscopy (DWS). Two major ways of modeling the propagation of coherent polarized light in scattering tissue-like media are currently in use. The first approach is tracking transformations of the electric field along ray propagation. Second one is developed in analogy to the iterative procedure of the solution of Bethe-Salpeter equation. In the current paper we compare these two approaches that have been extensively used in the past for simulation of coherent polarized light propagation in scattering tissue-like media, and quantitative assessment of the enhancement of coherent backscattering of light. In particular we compare the accuracy of each technique with the results obtained in experiments and with the results of known analytical solutions. The advantages and disadvantages of each technique and their further developments are discussed.

High scattering in biological turbid media limits the applicability of optical imaging techniques, such as optical coherence tomography (OCT). Accurate and robust simulations are required due to the complexity of optical wave propagation in these tissues. Recent computational simulations make use of finite-difference time-domain (FDTD) method to exactly solve the scattered electromagnetic field distribution. We propose a method to isolate and selectively remove energy from portions of the scattered field in these simulations. This technique will involve placing an absorber in the medium that consists of convolutional perfectly matched layers (CPML). The performance of the absorber as an optical target is analyzed.

This work presents an analytical approach for the solution of the tissue diffusion equation based on the bound- ary measurements. We consider a bioluminescent point source in both homogeneous and heterogeneous circular turbid media. The point source is described by the Dirac delta function. Analytical expressions for the strength and position of the point source are obtained introducing boundary measurements and then applying appropriate boundary conditions. In addition, numerical simulations are performed for the position of the source. Calculations show that that the analytical results are in a good accordance with the numerical results.

Fiber based frequency-modulated light scattering interferometry (FMLSI) is developed for optical properties studies of liquid phantoms, made of Intralipid®. By employing optical frequency modulation on a tunable diode laser, the power spectrum of the heterodyne-detected intensity fluctuations through the dynamic turbid medium is a combination of the time-of-flight distribution and the Doppler power spectrum due to the movement of the scattering particles. The reduced scattering coefficient, absorption coefficient and Brownian diffusion constant are retrieved by employing nonlinear fitting to the power spectrum based on diffusion theory.

We present an in vivo implementation of a multi-parametric technique for detecting apoptosis using optical coherence tomography in a mouse tumor model. Solid tumors were grown from acute myeloid leukemia cells in the hind leg of SCID mice and treated with a single dose of cisplatin and dexamethasone to induce apoptosis. Both spectral features and speckle decorrelation times indicated good consistency between control mice and reasonable agreement with in vitro measurements. The integrated backscatter increased significantly in tumors responding to treatment while the spectral slope and decorrelation time did not show significant changes. This study demonstrates the feasibility of using spectroscopic OCT and dynamic light scattering for treatment monitoring in vivo.

Cervical cancer is the eleventh most common cancer in the UK, especially for women under 35. In developed countries, cervical cancer is diagnosed by performing colposcopy. Contrast is enhanced by spraying dilute acetic acid onto the surface of the tissue. In the past decades, it has been shown that abnormal cervical epithelium turns opaque white upon contact with this weak acid whereas normal epithelium is generally not affected. This mechanism is known as aceto-whitening. However, the exact mechanism of this phenomenon is not fully known. In this study, OCT using near infrared light was used to quantify depth-resolved kinetics of aceto-whitening in a simple squamous epithelium model: rabbit cornea. We have found that both the epithelium and stroma brighten with approximately the same time course, reaching a peak reflectivity at about 50 seconds. The most significant increase in reflectivity was seen in the first 20 seconds upon the application of acid, and was measured to be 11dB. This result is compared with phosphate buffered saline solution, which was shown to exhibit no effect. Lactic acid, an alpha-hydroxy acid, has been reported as a negative control for aceto-whitening. However, our OCT results showed a significant epithelial brightening effect of approximately 8 dB in the first 20 seconds. The key difference with acetic acid is the lack of brightening in the corneal stroma. This could be due to inability to permeate through the basal lamina between corneal epithelium and stroma or lack of interaction with stromal keratocytes.

Dysplastic progression in epithelial tissues is linked to changes in morphology and internal structure of cell nuclei. These changes lead to alterations in nuclear light scattering profiles that can potentially be monitored for diagnostic purposes. Numerical tools allow for simulation of complex nuclear models and are particularly useful for quantifying the optical response of cell nuclei as dysplasia progresses. In this study, we first analyze a set of quantitative histopathology images from twenty cervical biopsy sections stained with Feulgen-thionin. Since Feulgen-thionin is stoichiometric for DNA, the images enable us to obtain detailed information on size, shape, and chromatin content of all the segmented nuclei. We use this extensive data set to construct realistic three-dimensional computational models of cervical cell nuclei that are representative of four diagnostic categories, namely normal or negative for dysplasia, mild dysplasia, moderate dysplasia, and severe dysplasia or carcinoma in situ (CIS). We then carry out finite-difference time-domain simulations to compute the light scattering response of the constructed models as a function of the polar scattering angle and the azimuthal scattering angle. The results show that these two-dimensional scattering patterns exhibit characteristic intensity ridges that change form with progression of dysplasia; pattern processing reveals that Haralick features can be used to distinguish moderately and severely dysplastic or CIS nuclei from normal and mildly dysplastic nuclei. Our numerical study also suggests that different angular ranges need to be considered separately to fully exploit the diagnostic potential of two-dimensional light scattering measurements.

When light interacts with a scattering medium, the spectrum of the incident light undergoes changes that are dependent on the size of the scatterers in the medium. Spectroscopic Optical Coherence Tomography (S-OCT) is a method that can be used to ascertain the resulting spatially-dependent spectral information. In fact, S-OCT is sensitive to structures that are below the spatial resolution of the system, making S-OCT a promising tool for diagnosing many diseases and biological processes that change tissue structure, like cancer. The most important signal processing steps for S-OCT are the depth-resolved spectral analysis and the calculation of a spectroscopic metric. While the former calculates the spectra from the raw OCT data, the latter analyzes the information content of the processed depth-resolved spectra. We combine the Dual Window spectral analysis with different spectroscopic metrics, which are used as an input to colorize intensity based images. These metrics include the spectral center of mass method, principal component (PCA) and phasor analysis. To compare the performance of the metrics in a quantitative manner, we use a cluster algorithm to calculate efficiencies for all methods. For this purpose we use phantom samples which contain areas of microspheres of different sizes. Our results demonstrate that PCA and phasor analysis have the highest efficiencies, and can clearly separate these areas. Finally we will present data from cartilage tissue under static load in vitro. These preliminary results show that S-OCT can generate additional contrast in biological tissue in comparison to the pure intensity based images.

The paper presents a new method for distant non-destructive determination of concentration of light absorbing admixtures in turbid media. In particular, it is intended for non-invasive in vivo control of accumulation in patient tissues of various biochemicals introduced to the patients for chemotherapy, photodynamic therapy or diagnostics. It is require that the admixture absorption spectrum should have a clearly marked peak in the wavelength region where the pure medium one varies regularly. Fluorescence of admixtures is not required. The method uses the local diffuse reflectance spectroscopy with optical fiber probe including one emitting and two reading There are several features in the method: the value to be determined is absolute concentration of admixtures; the method needs no calibration measurements on phantoms; it needs no reference measurements on sample with zero admixture concentration; it uses a two parametric kinetic light propagation model and original algorithms to resolve direct and inverse tasks of radiation transport theory. Experimental testing passed with tissue equivalent phantoms and different admixtures, including a chlorine photosensitizer, showed accuracy under 10% in all cases.

Laminar optical tomography (LOT) is a new mesoscopic functional optical imaging technique, which is an extension of a confocal microscope and diffuse optical tomography to acquire both the coaxial and off-axis scattered light at the same time. In this paper, a LOT system with a larger detection area aiming at the in vivo detection of early cervical cancer is developed. The field of view of our system is 10 mm x 10 mm. In order to improve the image quality of the system, two methods were performed: the correction of image distortion and the restriction of returning light. The performance of the system with aperture stop was assessed by liquid phantom experiments. Comparing with the Monte Carlo simulation, the measurement results show that the average relative errors of eight different source-detector distances corresponding to 4 source points are lower than the errors of the system taking the frame of objective lens as the aperture stop by 5.7%， 4.8%，6.1%，6.1% respectively. Moreover, the experiment based on the phantom with specified structure and optical parameters to simulate the cervix demonstrates that the system perform well for the cervix measurement.

Radiance is sensitive to the variations of tissue optical parameters, such as absorption coefficient μ_a, scattering coefficient μ_s, and anisotropy factor g. Therefore, similar to fluence, radiance can be used for tissue characterization. Compared with fluence, radiance has the advantage of offering the direction information of light intensity. Taking such advantage, the optical parameters can be determined by rotating the detector through 360 deg with only a single optode pair. Instead of the translation mode used in the fluence-based technologies, the Rotation mode has less invasiveness in the clinical diagnosis. This paper explores a new method to obtain the optical properties by measuring the distribution of light intensity in liquid phantom with only a single optode pair and the detector rotation through 360 deg. The angular radiance and distance-dependent radiance are verified by comparing experimental measurement data with Monte Carlo (MC) simulation for the short source-detector separations and diffusion approximation for the large source-detector separations. Detecting angular radiance with only a single optode pair under a certain source-detection separation will present a way for prostate diagnose and light dose calculation during the photon dynamic therapy (PDT).

According to the morphological differences in the vascularization between healthy and diseased tissues, pharmacokinetic-rate images of fluorophore can provide diagnostic information for tumor differentiation, and especially have the potential for staging of tumors. In this paper, fluorescence diffuse optical tomography method is firstly used to acquire metabolism-related time-course images of the fluorophore concentration. Based on a two-compartment model comprised of plasma and extracelluar-extravascular space, we next propose an adaptive-EKF framework to estimate the pharmacokinetic-rate images. With the aid of a forgetting factor, the adaptive-EKF compensate the inaccuracy initial values and emphasize the effect of the current data in order to realize a better online estimation compared with the conventional EKF. We use simulate data to evaluate the performance of the proposed methodology. The results suggest that the adaptive-EKF can obtain preferable pharmacokinetic-rate images than the conventional EKF with higher quantitativeness and noise robustness.

To realize three-dimensional (3D) optical imaging of the internal structure of animal body, we have developed a new technique to reconstruct CT images from two-dimensional (2D) transillumination images. In transillumination imaging, the image is blurred due to the strong scattering in the tissue. We had developed a scattering suppression technique using the point spread function (PSF) for a fluorescent light source in the body. In this study, we have newly proposed a technique to apply this PSF for a light source to the image of unknown light-absorbing structure. The effectiveness of the proposed technique was examined in the experiments with a model phantom and a mouse. In the phantom experiment, the absorbers were placed in the tissue-equivalent medium to simulate the light-absorbing organs in mouse body. Near-infrared light was illuminated from one side of the phantom and the image was recorded with CMOS camera from another side. Using the proposed techniques, the scattering effect was efficiently suppressed and the absorbing structure can be visualized in the 2D transillumination image. Using the 2D images obtained in many different orientations, we could reconstruct the 3D image. In the mouse experiment, an anesthetized mouse was held in an acrylic cylindrical holder. We can visualize the internal organs such as kidneys through mouse’s abdomen using the proposed technique. The 3D image of the kidneys and a part of the liver were reconstructed. Through these experimental studies, the feasibility of practical 3D imaging of the internal light-absorbing structure of a small animal was verified.

Mueller matrix imaging techniques can be used to detect the micro-structure variations of superficial biological tissues, including the sizes and shapes of cells, the structures in cells, and the densities of the organelles. Many tissues contain anisotropic fibrous micro-structures, such as collagen fibers, elastin fibers, and muscle fibers. Changes of these fibrous structures are potentially good indicators for some pathological variations. In this paper, we propose a quantitative analysis technique based on Mueller matrix for mapping local anisotropy axis of scattering media. By conducting both experiments on silk sample and Monte Carlo simulation based on the sphere-cylinder scattering model (SCSM), we extract anisotropy axis parameters from different backscattering Mueller matrix elements. Moreover, we testify the possible applications of these parameters for biological tissues. The preliminary experimental results of human cancerous samples show that, these parameters are capable to map the local axis of fibers. Since many pathological changes including early stage cancers affect the well aligned structures for tissues, the experimental results indicate that these parameters can be used as potential tools in clinical applications for biomedical diagnosis purposes.

In this paper, we study the cylindrical scatterers and birefringent medium as two anisotropic sources in the turbid medium by polarimetry method. Using Monte Carlo simulations on the anisotropic sample, we examine in detail the relationship between the retardance δ, depolarization Δ and the anisotropic microscopic structure and their interactions of each other by Mueller matrix decomposition method. The well-ordered cylinder scatter system can be regarded as a birefringent medium. If the orientation of the aligned cylindrical scatterers does not coincide with the extraordinary axis of the birefringent medium, the variation of total retardance in this cylinder-birefringence model would accord to the two pieces of birefringent medium overlapped together with angle between extraordinary axes.

Measuring microcirculatory tissue blood perfusion is of interest for both clinicians and researchers in a wide range of applications and can provide essential information of the progress of treatment of certain diseases which causes either an increased or decreased blood flow. Diabetic ulcer associated with alterations in tissue blood flow is the most common cause of non-traumatic lower extremity amputations. A technique which can detect the onset of ulcer and provide essential information on the progress of the treatment of ulcer would be of great help to the clinicians. A noninvasive, noncontact and whole field laser speckle contrast imaging (LSCI) technique has been described in this paper which is used to assess the changes in blood flow in diabetic ulcer affected areas of the foot. The blood flow assessment at the wound site can provide critical information on the efficiency and progress of the treatment given to the diabetic ulcer subjects. The technique may also potentially fulfill a significant need in diabetic foot ulcer screening and management.