This PDF file contains the front matter associated with SPIE Proceedings Volume 8942, including the Title Page, Copyright Information, Table of Contents, and the Conference Committee listing.

The ability to measure polarization effects is important in many biological and industrial applications. Additionally, measuring spectral and scatter effects can offer greater sensitivity for applications where characterization and differentiation are important. Here we present a liquid crystal-based spectral imaging goniometric polarimeter to probe these effects. The system consists of two modules, a Stokes generator and a polarimeter, each constructed from a pair of liquid crystal variable retarders (LCVR). LCVRs are computer-controlled birefringent devices that impart retardance effects with no mechanical movement. Additionally, the Stokes generator utilizes a computer-controlled liquid crystal tunable filter (LCTF) to transmit a specific wavelength bandwidth, also with no mechanical movement. The polarimeter, enclosed in a cage system, manually rotates around the sample plane to provide angular scatter measurements. A CCD camera images the sample and provides spatially resolved estimates of the complete Mueller matrix as a function of wavelength and scatter angle. Here we describe the system and its calibration, and show quantitative measurement results for a number of samples.

Multiple reference optical coherence tomography (MR-OCT) is a recently developed time-domain interferometeric imaging platform, which promises to fit into robust, cost-effective designs that are virtually solid state. An optical delay is created by the reference mirror which is mounted on a piezo-electric transducer (PZT) and the key element of MR-OCT technology is the presence of a partial mirror in front of the reference mirror. However, the limited axial displacement range at higher scanning-frequencies are a limitation of a PZT-based optical delay. Moreover, PZT-based actuators require a relatively high operational voltage and are expensive. In this paper we present a voice coil actuator as an alternative to a PZT-based optical delay. Voice coil actuators are light in weight, inexpensive and offer other advantages such as zero hysteresis, low operational voltage and a long life. We demonstrate a voice coil actuator as a feasible alternative to PZT-based actuators for the purpose of creating an optical delay, which can provide fast and precise axial displacements at high scanning rates.

Optical coherence tomography (OCT) is an optical imaging technique that is capable of performing high-resolution (approaching the histopathology level) and real-time imaging of tissues without use of contrast agents. Based on these advantages, the pathological features of tumors (in micro scale) can be identified during resection surgery. However, the accuracy of tumor margin prediction still needs to be enhanced for assisting the judgment of surgeons. In this regard, we present a two-dimensional computational method for advanced tissue analysis and characterization based on optical coherence tomography (OCT) and Raman spectroscopy (RS). The method combines the slope of OCT intensity signal and the Principal component (PC) of RS, and relies on the tissue optical attenuation and chemical ingredients for the classification of tissue types. Our pilot experiments were performed on mouse kidney, liver and small intestine. Results demonstrate the improvement of the tissue differentiation compared with the analysis only based on the OCT detection. This combined OCT/RS method is potentially useful as a novel optical biopsy technique for cancer detection.

Within the field of tissue engineering there is an emphasis on studying 3-D live tissue structures. Consequently, to investigate and identify cellular activities and phenotypes in a 3-D environment for all in vitro experiments, including shape, migration/proliferation and axon projection, it is necessary to adopt an optical imaging system that enables monitoring 3-D cellular activities and morphology through the thickness of the construct for an extended culture period without cell labeling. This paper describes a new 3-D tracking algorithm developed for Cell-IQ®, an automated cell imaging platform, which has been equipped with an environmental chamber optimized to enable capturing time-lapse sequences of live cell images over a long-term period without cell labeling. As an integral part of the algorithm, a novel auto-focusing procedure was developed for phase contrast microscopy equipped with 20x and 40x objectives, to provide a more accurate estimation of cell growth/trajectories by allowing 3-D voxels to be computed at high spatiotemporal resolution and cell density. A pilot study was carried out in a phantom system consisting of horizontally aligned nanofiber layers (with precise spacing between them), to mimic features well exemplified in cellular activities of neuronal growth in a 3-D environment. This was followed by detailed investigations concerning axonal projections and dendritic circuitry formation in a 3-D tissue engineering construct. Preliminary work on primary animal neuronal cells in response to chemoattractant and topographic cue within the scaffolds has produced encouraging results.

Non-contact based near-infrared (NIR) optical imaging devices are developed for non-invasive imaging of deep tissues in various clinical applications. Most of these devices focus on obtaining the spatial information for anatomical co-registration of blood vessels as in sub-surface vein localization applications. In the current study, the anatomical co-registration of blood vessels based on spatio-temporal features was performed using NIR optical imaging without the use of external contrast agents. A 710 nm LED source and a compact CCD camera system were employed during simple cuff (0 to 60 mmHg) experiment in order to acquire the dynamic NIR data from the dorsum of a hand. The spatio-temporal features of dynamic NIR data were extracted from the cuff experimental study to localize vessel according to blood dynamics. The blood vessels shape is currently reconstructed from the dynamic data based on spatio-temporal features. Demonstrating the spatio-temporal feature of blood dynamic imaging using a portable non-contact NIR imaging device without external contrast agents is significant for applications such as peripheral vascular diseases.

Cutaneous wound healing consists of multiple overlapping phases starting with blood coagulation following incision of blood vessels. In this paper, we briefly review wound healing phases that were observed by utilizing optical microangiography (OMAG) to monitor healing process and dynamics of microcirculation system in a mouse ear pinna wound model. Mouse ear pinna is composed of two layers of skin separated by a layer of cartilage and because its total thickness is around 500 μm, can be utilized as an ideal model for optical imaging techniques. These skin layers are identical to human skin structure except for sweat ducts and glands. Microcirculatory system responds to the wound injury by recruiting collateral vessels to supply blood flow to hypoxic area. Also, lymphatic vessels play an important role in the immune response of the tissue and clearing waste from interstitial fluid.

The objective of the present work is to diagnose pre-cancer by wavelet transform and multi-fractal de-trended fluctuation analysis of DIC images of normal and different grades of cancer tissues. Our DIC imaging and fluctuation analysis methods (Discrete and continuous wavelet transform, MFDFA) confirm the ability to diagnose and detect the early stage of cancer in cervical tissue.

In this work photoacoustic imaging (PAI) based on multi element linear-array transducer, combined with multichannel collecting system was used for in vivo imaging of microcirculation of the human forearm. The Vevo® 2100 LAZR PAT system (VISUALSONICS) was used for imaging which simultaneously collects high-resolution ultrasound and photoacoustic signals. 3D PA and high frequency ultrasound scans, measured 30.5 mm (length) x 14.1 mm (width) x 10 mm (depth) were acquired from the area of forearm skin using 40 MHz frequency transducer at 860 nm wavelength. 3D structural and functional (microcirculation) maps of the forearm skin were obtained. The multi element linear-array transducer based PAI has been found promising in terms of resolution, imaging depth and imaging speed for in vivo microcirculation imaging within human skin.

Choroidal thickness (ChT), defined as the distance between the retinal pigment epithelium (RPE) and the choroid-sclera interface (CSI), is highly correlated with various ocular disorders like high myopia, diabetic retinopathy, and central serous chorioretinopathy. Long wavelength Optical Coherence Tomography (OCT) has the ability to penetrate deep to the CSI, making the measurement of the ChT possible. The ability to accurately segment the CSI and RPE is important in extracting clinical information. However, automated CSI segmentation is challenging due to the weak boundary in the lower choroid and inconsistent texture with varied blood vessels. We propose a K-means clustering based automated algorithm, which is effective in segmenting the CSI and RPE. The performance of the method was evaluated using 531 frames from 4 normal subjects. The RPE and CSI segmentation time was about 0.3 seconds per frame, and the average time was around 0.5 seconds per frame with correction among frames, which is faster than reported algorithms. The results from the proposed method are consistent with the manual segmentation results. Further investigation includes the optimization of the algorithm to cover more OCT images captured from patients and the increase of the processing speed and robustness of the segmentation method.

Ability to non-invasively monitor and quantify of blood flow, blood vessel morphology, oxygenation and tissue morphology is important for improved diagnosis, treatment and management of various neurovascular disorders, e.g., stroke. Currently, no imaging technique is available that can satisfactorily extract these parameters from in vivo microcirculatory tissue beds, with large field of view and sufficient resolution at defined depth without any harm to the tissue. In order for more effective therapeutics, we need to determine the area of brain that is damaged but not yet dead after focal ischemia. Here we develop an integrated multi-functional imaging system, in which SDW-LSCI (synchronized dual wavelength laser speckle imaging) is used as a guiding tool for OMAG (optical microangiography) to investigate the fine detail of tissue hemodynamics, such as vessel flow, profile, and flow direction. We determine the utility of the integrated system for serial monitoring afore mentioned parameters in experimental stroke, middle cerebral artery occlusion (MCAO) in mice. For 90 min MCAO, onsite and 24 hours following reperfusion, we use SDW-LSCI to determine distinct flow and oxygenation variations for differentiation of the infarction, peri-infarct, reduced flow and contralateral regions. The blood volumes are quantifiable and distinct in afore mentioned regions. We also demonstrate the behaviors of flow and flow direction in the arterials connected to MCA play important role in the time course of MCAO. These achievements may improve our understanding of vascular involvement under pathologic and physiological conditions, and ultimately facilitate clinical diagnosis, monitoring and therapeutic interventions of neurovascular diseases, such as ischemic stroke.

We use rodent parietal cortex as a model system and utilize a synchronized dual wavelength laser speckle imaging (SDW-LSCI) technique to explore the hemodynamic response of infarct and penumbra to a brain injury (middle cerebral artery occlusion (MCAO) model). The SDW-LSCI system is able to take snapshots rapidly (maximum 500 Hz) over the entire brain surface, providing key information about the hemodynamic response, in terms of which it may be used to elucidate evolution of penumbra region from onsite to 90 min of MCAO. Changes in flow are quantified as to the flow experiencing physical occlusions of the MCA normalized to that of baseline. Furthermore, the system is capable of providing information as to the changes of the concentration of oxygenated, (HbO) deoxygenated (Hb), and total hemoglobin (HbT) in the cortex based on the spectral characteristics of HbO and Hb. We observe that the oxygenation variations in the four regions are detectable and distinct. Combining the useful information, four regions of interest (ROI), infarct, penumbra, reduced flow and contralateral portions in the brain upon ischemic injury may be differentiated. Implications of our results are discussed with respect to current understanding of the mechanisms underlying MCAO. We anticipate that SDW-LSCI holds promise for rapid and large field of view localization of ischemic injury.

Cellular adhesions play a critical role in cell behavior, and modified expression of cellular adhesion compounds has been linked to various cancers. We tested the role of cellular adhesions in drug response by studying three cellular culture models: three-dimensional tumor spheroids with well-developed cellular adhesions and extracellular matrix (ECM), dense three-dimensional cell pellets with moderate numbers of adhesions, and dilute three-dimensional cell suspensions in agarose having few adhesions. Our technique for measuring the drug response for the spheroids and cell pellets was biodynamic imaging (BDI), and for the suspensions was quasi-elastic light scattering (QELS). We tested several cytoskeletal chemotherapeutic drugs (nocodazole, cytochalasin-D, paclitaxel, and colchicine) on three cancer cell lines chosen from human colorectal adenocarcinoma (HT-29), human pancreatic carcinoma (MIA PaCa-2), and rat osteosarcoma (UMR-106) to exhibit differences in adhesion strength. Comparing tumor spheroid behavior to that of cell suspensions showed shifts in the spectral motion of the cancer tissues that match predictions based on different degrees of cell-cell contacts. The HT-29 cell line, which has the strongest adhesions in the spheroid model, exhibits anomalous behavior in some cases. These results highlight the importance of using three-dimensional tissue models in drug screening with cellular adhesions being a contributory factor in phenotypic differences between the drug responses of tissue and cells.

The electric field induced optical changes (EIOC) measured by the optical coherence tomography (OCT) reflect the local electro-kinetic properties of the tissue. In this study we developed a method to use the phase of the complex OCT images to map EIOC in tissue samples. Switching the polarity of the electric field induced significant reversible changes in the phase of the complex OCT images. Since the resulting phase was degraded by the noise an advanced signal processing algorithm was developed to obtain the EIOC images. The developed algorithm made it possible to get structural phase images from a standard commercial OCT system, potentially yielding important insights into the local electro-kinetic properties of the tissue. We use a simple theoretical model to simulate main features of amplitude and phase EIOC observed using frequency-domain OCT.

NIRS analysis is considered to be a promising noninvasive detection technique. Existing research results show that optical properties of the human skin tissue will change with different contact pressures when contact survey mode is used for in vivo NIRS measurement. The impact of the contact pressure might be greater than the impact of glucose concentration on the spectral data of NIRS. The uncertainty caused by pressure in vivo, makes it extremely difficult to get the high SNR-spectrum. In this talk, the Monte Carlo simulation has been carried on under the condition of different contact pressure. Simulation results show that the diffused reflectance and transmittance increase with rising contact pressure. Moreover, the simulation results show that the effects of contact pressure are larger than the effects of glucose concentration. Therefore it is promising to make comprehensive utilization of diffused reflectance and transmittance to eliminate the interference which caused by contact pressure.

Scale Invariant Feature Transform（SIFT）algorithm is widely used for ear feature matching and recognition. However, the application of the algorithm is usually interfered by the non-target areas within the whole image, and the interference would then affect the matching and recognition of ear features. To solve this problem, a combined image segmentation algorithm i.e. KRM was introduced in this paper, As the human ear recognition pretreatment method. Firstly, the target areas of ears were extracted by the KRM algorithm and then SIFT algorithm could be applied to the detection and matching of features. The present KRM algorithm follows three steps: (1)the image was preliminarily segmented into foreground target area and background area by using K-means clustering algorithm; (2)Region growing method was used to merge the over-segmented areas; (3)Morphology erosion filtering method was applied to obtain the final segmented regions. The experiment results showed that the KRM method could effectively improve the accuracy and robustness of ear feature matching and recognition based on SIFT algorithm.

Hair follicle, as a mini-organ with perpetually cycling of telogen, anagen and catagen, provides a valuable experimental model for studying hair and organ regeneration. The transition of hair follicle from telogen to anagen is a significant sign for successful regeneration. So far discrimination of the hair follicle stage is mostly based on canonical histological examination and empirical speculation based on skin color. Hardly a method has been proposed to quantitatively evaluate the hair follicle stage. In this work, a commercial optical fiber spectrometer was applied to monitor diffuse reflectance of mouse skin with hair follicle cycling, and then the change of reflectance was obtained. Histological examination was used to verify the hair follicle stage. In comparison with the histological examination, the skin diffuse reflectance was relatively high for mouse with telogen hair follicles; it decreased once hair follicles transited to anagen stage; then it increased reversely at catagen stage. This study provided a new method to quantitatively evaluate the hair follicle stage, and should be valuable for the basic and therapeutic investigations on hair regeneration.

Photodynamic inactivation of some microorganisms (St. aureus, E.coli) was investigated and their dependence on photo-physical properties of photosensitizers (PS) (cationic porphyrins and metalloporphyrins) was shown. One of the most important criteria for the effectiveness of the PS`s is the quantum yield of singlet oxygen (γ_Δ). Our investigations were shown that γ_Δ of metalloporphyrins, containing Zn, significantly higher than of metal-free porphyrins (85-97% and 77-79%, respectively). Previousl y experimentally we were found that under the action cationic porphyrins and metalloporphyrins on Gram (+) and Gram (-) microorganisms efficiency of metalloporphyrins Zn-TOEt4PyP and Zn-TBut4PyP in 3-5 times was higher than the metal-free porphyrins. In this study under the action of porphyrins and their Zn-derivatives on microorganism St. aureus such an effect was confirmed. Using the LED with a peak emission of 405 nm and a power density of 70 mW/cm², and irradiation time of microorganisms from 5 to 30 minutes we have found, that at a concentration of 0.1 ug/ml the highest efficiency is observed of metalloporphyrin Zn-TBut3PyP. Upon irradiation of 10 and 15 min his efficiency is 3-5 times higher than the metal-free porphyrin TOEt4PyP, and irradiation for 30 min via Zn-TBut3PyP is practically completely kills microorganisms. These data correlate with the quantum yield of singlet oxygen for photosensitizers. The 30 mindirect sun exposure (power density of 70 mW/cm²) of photosensitizer solutions showed that a significant photobleaching of porphyrins and metalloporphyrins does not occur. Thus, Zn-containing cationic metalloporphyrins are highly efficient photosensitizers for photodynamic inactivation of microorganisms and PDT.