This PDF file contains the front matter associated with SPIE Proceedings Volume 9707, including the Title Page, Copyright information, Table of Contents, Introduction, and the Conference Committee listing.

Laser speckle Micro-rheology (LSM) is a novel optical tool for evaluating the viscoelastic properties of biomaterials. In LSM, a laser beam illuminates the specimen and scattered rays are collected through an objective by a high-speed CMOS camera. The self-interference of light rays forms a fluctuating speckle pattern captured by the CMOS sensor. Spatio-temporal correlation analysis of speckle images provides the intensity autocorrelation function, g2(t), for individual pixels. Next, the mean square displacements (MSD) of Brownian particles are deduced and substituted in the generalized Stokes-Einstein relation (GSER) to yield a 2D map of viscoelastic modulus, |G*(ω)|. To compare the accuracy, sensitivity, and dynamic range of LSM measurements with standard mechanical testing methods, homogeneous polyethylene glycol (PEG), agarose, and polyacrylamide (PA) gels, of assorted viscoelastic properties were fabricated and evaluated using LSM, shear rheology, and indentation-mode atomic force microscopy (AFM). Results showed a statistically significant, strong correlation between G* values measured by LSM and shear rheology (R=0.94, p<5x10-6) (|G*|: 30 Pa - 30 kPa at ω = 1 Hz). Likewise, strong correlation was observed between G* values measured by LSM and indentation moduli of AFM (R=0.94, p,0.05). Next, polyacrylamide substrates with micro-scale stiffness patterns were tested using LSM. The reconstructed |G*| maps illustrated the high sensitivity of LSM in resolving mechanical heterogeneities below 100 microns. These findings demonstrate the competent accuracy and sensitivity of LSM measurements. Moreover, the non-contact nature of LSM provides a major advantage over mechanical tests, making it suitable for in vivo studies in future.

Laser speckle imaging (LSI) enables measurement of relative blood flow in microvasculature and perfusion in tissues. To determine the impact of tissue optical properties and perfusion dynamics on speckle contrast, we developed a computational simulation of laser speckle contrast imaging. We used a discrete absorption-weighted Monte Carlo simulation to model the transport of light in tissue. We simulated optical excitation of a uniform flat light source and tracked the momentum transfer of photons as they propagated through a simulated tissue geometry. With knowledge of the probability distribution of momentum transfer occurring in various layers of the tissue, we calculated the expected laser speckle contrast arising with coherent excitation using both reflectance and transmission geometries. We simulated light transport in a single homogeneous tissue while independently varying either absorption (.001-100mm^-1), reduced scattering (.1-10mm^-1), or anisotropy (0.05-0.99) over a range of values relevant to blood and commonly imaged tissues. We observed that contrast decreased by 49% with an increase in optical scattering, and observed a 130% increase with absorption (exposure time = 1ms). We also explored how speckle contrast was affected by the depth (0-1mm) and flow speed (0-10mm/s) of a dynamic vascular inclusion. This model of speckle contrast is important to increase our understanding of how parameters such as perfusion dynamics, vessel depth, and tissue optical properties affect laser speckle imaging.

In this work the effects of incident intensity and effective camera dynamic range on image acquisition of both frozen and time-averaged dynamic speckle patterns, and their effects on laser speckle contrast imaging are addressed. A nematic liquid crystal, phase-only, spatial light modulator (SLM) was employed to generate laser speckle in a controlled and repeatable fashion. By addressing the calculated spatial contrast of frozen and time-averaged dynamic speckle patterns imaged across a wide range of intensities, we present a description of optimum intensity characteristics that should be observed when using LSCI. The results indicate the importance of assessing the intensity of the signal quantized by the camera in LSCI. By analyzing intensity PDF’s during image acquisition of speckle patterns used in LSCI, an optimum incident intensity can be detected when a single, polarized speckle frame displays the first order statistics characteristic of fully developed speckle. Our results indicate that there is a range of laser power densities where the ensuing imaged speckle exhibit optimum sensitivity to flow as well as relatively constant calculated contrast values. It is clear that at high intensities, high frequency information is lost due to camera saturation, resulting in a decrease in contrast. When imaging speckle at low intensity, there is a risk for loss of data during the digital quantization process. The results are presented in a generalized fashion, so they should be applicable to any LSCI system, regardless of incident laser power or camera depth.

It is widely recognized that effective stress management could have a dramatic impact on health care and preventive medicine. In order to meet this need, efficient and seamless sensing and analytic tools for the non-invasive stress monitoring during daily life are required. The existing sensors still do not meet the needs in terms of specificity and robustness. We utilized a miniaturized dynamic light scattering sensor (mDLS) which is specially adjusted to measure skin blood flow fluctuations and provides multi- parametric capabilities. Based on the measured dynamic light scattering signal from the red blood cells flowing in skin, a new concept of hemodynamic indexes (HI) and oscillatory hemodynamic indexes (OHI) have been developed. This approach was utilized for stress level assessment for a few usecase scenario. The new stress index was generated through the HI and OHI parameters. In order to validate this new non-invasive stress index, a group of 19 healthy volunteers was studied by measuring the mDLS sensor located on the wrist. Mental stress was induced by using the cognitive dissonance test of Stroop. We found that OHIs indexes have high sensitivity to the mental stress response for most of the tested subjects. In addition, we examined the capability of using this new stress index for the individual monitoring of the diurnal stress level. We found that the new stress index exhibits similar trends as reported for to the well-known diurnal behavior of cortisol levels. Finally, we demonstrated that this new marker provides good sensitivity and specificity to the stress response to sound and musical emotional arousal.

Currently the diagnosis of hemorrhagic shock is essentially clinical, relying on the expertise of nurses and doctors. One of the first measurable physiological changes that marks the onset of hemorrhagic shock is a decrease in capillary blood flow. Diffuse correlation spectroscopy (DCS) quantifies this decrease. DCS collects and analyzes multiply scattered, coherent, near infrared light to assess relative blood flow. This work presents a preliminary study using a DCS instrument with human subjects undergoing a lower body negative pressure (LBNP) protocol. This work builds on previous successful DCS instrumentation development and we believe it represents progress toward understanding how DCS can be used in a clinical setting.

We present an optical coherence tomography (OCT) based method for mapping transverse red blood cell (RBC) flow speed at capillary. This OCT velocimetry utilizes a quantitative laser speckle temporal contrast analysis that estimates reliable speckle decorrelation time from the observed speckle contrast, which is related to microcirculatory flow velocity. For capillary speed measurement, we employ a home-built 1.3 µm MHz swept-source OCT (SS-OCT) system that can acquire OCT B-frames at a rate of 1.7 kHz. From the multiple B-frames obtained at the same location, intensity profiles with time-varying OCT speckle contrast are extracted at single capillaries using a capillary binary mask and then the transverse flow speed is calculated by adapting the profiles to the speckle contrast analytic model. Finally, a 3D speed map can be achieved for OCT volume imaging. To validate this method, we perform a systematic study using both phantom and in vivo rodent models. Result shows that our method is effective to measure transverse capillary flow speed.

In this study, we present a technique to image the enhanced particle displacement generated using an acoustic radiation force (ARF) excitation source. A swept-source OCT (SS-OCT) system with a center wavelength of 1310nm, a bandwidth of ~100nm, and an A-scan rate of 100 kHz (MEMS-VCSEL OCT Thorlabs) was used to detect gold nanoparticle (70nm in diameter) displacement .ARF was applied after the nanoparticles passed through a porous membrane and diffused into a collagen (6% collagen) matrix. B-mode, M-B mode, 3D and Speckle Variance (SV) images were acquired before and after the ARF beam was on. Differential OCT speckle variance images with and without the ARF were used to measure the particle displacement. The images were used to detect the microscopic enhancement of nanoparticle displacement generated by the ARF. Using this OCT imaging technique, the extravasation of particles though a porous membrane and characterization of the enhanced particle displacement in a collagen gel after using an ARF excitation was achieved.

We consider the mechanism of speckle patterns formation in time-domain and swept source optical coherence tomography (OCT), and introduce a Monte Carlo based model for simulating OCT signals and images. The model takes into account polarization and coherent properties of light, mutual interference of the back-scattering light, and its interference with the reference beam. The developed model is employed to generate OCT images, and to analyze the resultant OCT speckle pattern properties. The model simulation results are compared with experimental measurements, and an interpretation of the speckle patterns formation in terms of its underlying physics is provided.

Unilateral carotid artery ligation which could induce adaptive improvement is a classic model that has been widely used to study pathology of ischemic disease. In those studies, blood flow is an important parameter to characterize the ischemia. Optical coherence tomography (OCT) is a powerful imaging modality which can provide depth resolved images in biological tissue with high spatial and temporal resolution. SPF rats was anesthetized with isoflurane and divided into two groups. In first group, bilateral carotid artery was surgically exposed, and then left carotid artery was ligated. Blood flow changes of the contralateral carotid artery was monitored using high speed spectral domain optical coherence tomography, including the absolute flow velocity and the flow volume. In the other group, skull window was opened at the ipsilateral cerebral cortex of ligation and blood supply of small artery was measured before and after the ligation. The measured results demonstrate the blood supply compensation process after unilateral carotid artery ligation. With the superiority of high resolution, OCT is an effective technology in monitoring results of carotid artery after ligation.

We develop a modulus Mueller matrix microscope by adding polarization state generator and analyzer (PSG and PSA) to a commercial transmission optical microscope. The microscope is calibrated to minimize errors of the Mueller matrix elements to 0.01. Preliminary experimental results on biological tissues and different materials and Monte Carlo simulations show that the microscope can provide quantitative information on the characteristic microstructural features of complex samples, which make it a potentially powerful tool in clinical diagnosis and material studies.

Studying neurovascular blood flow function in cerebrovascular activities requires accurate visualization and characterization of blood flow volume as well as the dynamics of blood cells in microcirculation. In this study, we present a novel integration of laser speckle contrast imaging (LSCI) and spectral domain optical coherence tomography (SD-OCT) for rapid volumetric imaging of blood flow in cortical capillaries. LSCI uses the illumination of wide-field near infrared light (NIR) and monitors back scattered light to characterize the relative dynamics of blood flow in microcirculation. Absolute measurement of blood cells and blood volume requires high-resolution volumetric structural information. SD-OCT system uses coherence gating to measure scattered light from a small volume within high structural resolution. The structural imaging system rapidly assesses large number of capillaries for spatio-temporal tracking of red blood cells (RBC). A very fast-ultra resolution SD-OCT system was developed for imaging high-resolution volumetric samples. The system employed an ultra wideband light source (1310 ± 200 nm in wavelength) corresponding to an axial resolution of 3 micrometers in tissue. The spectrometer of the SD-OCT was customized for a maximum scanning rate of 147,000 line/s. We demonstrated a fast volumetric OCT angiography algorithm to visualize large numbers of vessels in a 2-mm deep sample volume. A LSCI system that has been developed previously in our group was integrated to the imaging system for the characterization of dynamic blood cells. The conjunction data from LSCI and SD-OCT systems imply the feasibility of accurate quantification of absolute cortical blood flow.

I will review recent advances and applications in Diffuse Correlation Spectroscopy, Laser Speckle Contrast Imaging, and Optical Coherence Tomography for measuring dynamics arising from cerebral blood flow and intra-cellular organelle motility.

We introduce a new software correlator approach for continuous high-speed (up to 100 Hz) monitoring of blood flow dynamics with Diffuse Correlation Spectroscopy. The functionality of the high-speed software correlator is demonstrated with measurements of baseline blood flow dynamics. The utility of high-data-rate blood flow monitoring is demonstrated with measurements of cerebral autoregulation dynamics.

The requirements on nanoparticles for cosmetic and medical applications are very different. While nanoparticles applied in sunscreens shall remain on the skin surface or in the upper cell layers of the stratum corneum, nanoparticles for medical drug delivery shall penetrate through the skin barrier to the target structures in the living cells. Under the Collaborative Research Project 1112 various methods are used at the CCP to investigate the cutaneous penetration and storage of nanoparticles, hair follicles being in the focus of attention. Human hair follicles are ideal target structures for drug delivery. Hosting both the stem and dendritic cells, they are surrounded by a dense network of blood vessels. Investigating nanoparticles of different size and materials, particles of approximately 600 nm in diameter were found to penetrate best into the hair follicles, where they can be stored for maximally 10 days. Their retention time in the hair follicles exceeds that in the stratum corneum by almost one order of magnitude. Particles penetrate more efficiently into the hair follicles than non-particulate substances. For particles from 40 nm-1 µm in diameter, however, no follicular penetration has been detectable if the skin barrier was intact. This is plausible as the hair follicle has its own barrier. It will be demonstrated that the best way for drug delivery is the application of drug-loaded particulate carrier systems. In the hair follicles the particles may either dissolve and release the drug, or an external signal must trigger the drug release from the particle.

Systemic sclerosis (SSc) is a connective tissue disease that results in excessive accumulation of collagen in the skin and internal organs. Overall, SSc is a rare disorder, but has a high mortality, particularly in last decade of life. To improve the survival rate, an accurate and early diagnosis is crucial. Currently, the modified Rodnan skin score (mRSS) is the gold standard for evaluating SSc progression based on clinical palpation at 17 sites on the body. However, this procedure can be time consuming, and the assessed score may be biased by the experience of the clinician, causing inter- and intraobserver variabilities. Moreover, the instrinsic elasticity of skin may further bias the mRSS assessment in the early stages of SSc, such as oedematous. To overcome these limitations, there is a need for a rapid, accurate, and objective assessment technique. Optical coherence elastography (OCE) is a novel, rapidly emerging technique, which can assess mechanical contrast in tissues with micrometer spatial resolution. In this work, we demonstrate the first use of OCE to assess the mechanical properties of control and SSc-like diseased skin non-invasively. A focused air-pulse induced an elastic wave in the skin, which was detected by a home-built OCE system. The elastic wave propagated significantly faster in SSc skin compared to healthy skin. The Young’s modulus of the SSc skin was significantly higher than that of normal skin (P<0.05). Thus, OCE was able to objectively differentiate healthy and fibrotic skin completely noninvasively and is a promising and potentially useful new technology for quantifying skin involvement in SSc.

Polarization images contain abundant microstructural information of samples. Recently, as a comprehensive description of the structural and optical properties of complex media, the Mueller matrix imaging has been widely applied to biomedical studies, especially cancer detections. In previous works, we proposed a technique to transform the backscattering 3 × 3 Mueller matrices into a group of quantitative parameters with clear relationships to specific microstructures. In this paper, we extend this transformation method to full 4 × 4 Mueller matrices of both the back and forward scattering directions. Using the experimental results of phantoms and Monte Carlo simulation based on the sphere-cylinder birefringence model, we fit the Mueller matrix elements to trigonometric curves in polar coordinates and obtain a new set of transformation parameters, which can be expressed as analytical functions of 16 Mueller matrix elements. Both the experimental and simulated results demonstrate that the transformation parameters have simple relationships to the characteristic microstructural properties, including the densities and orientations of fibrous structures, the sizes of the scatterers, and the depolarization power of the samples. We also apply the transformation parameters of full 4 × 4 Mueller matrices to human liver cancerous tissues. Preliminary imaging results show that the parameters can quantitatively reflect the formation of fibrous birefringent tissues accompanying the cancerous processes. The findings presented in this study can be useful for in vivo or in vitro polarization imaging of tissues for diagnostic applications.

A non-contact imaging method with a digital RGB camera is proposed to evaluate plethysmogram and spontaneous lowfrequency oscillation. In vivo experiments with human skin during mental stress induced by the Stroop color-word test demonstrated the feasibility of the method to evaluate the activities of autonomic nervous systems.

The results of measuring of biomarkers in breath air of patients with broncho-pulmonary diseases using wide-band frequency tuning IR laser photo-acoustic spectroscopy and the methods of data mining are presented. We will discuss experimental equipment and various methods of intellectual analysis of the experimental spectra in context of above task. The work was carried out with partial financial support of the FCPIR contract No 14.578.21.0082 (ID RFMEFI57814X0082).

Melanoma is an underserved area of cancer research, with little focus on studying the effects of tumor extracellular matrix (ECM) properties on melanoma tumor progression, metastasis, and treatment efficacy. We’ve developed a Raman spectral mapping-based in-vitro screening platform that allows for nondestructive in-situ, multi-time point assessment of a novel potential nanotherapeutic adjuvant, nanoceria (cerium oxide nanoparticles), for treating melanoma. We’ve focused primarily on understanding melanoma tumor ECM composition and how it influences cell morphology and ICC markers. Furthermore, we aim to correlate this with studies on nanotherapeutic efficacy to coincide with the goal of predicting and preventing metastasis based on ECM composition. We’ve compiled a Raman spectral database for substrates containing varying compositions of fibronectin, elastin, laminin, and collagens type I and IV. Furthermore, we’ve developed a machine learning-based semi-quantitative analysis platform utilizing dimensionality reduction with subsequent pixel classification and semi-quantitation of ECM composition using Direct Classical Least Squares for classification and estimation of the reorganization of these components by taking 2D maps using Raman spectroscopy. Gaining an understanding of how tissue properties influence ECM organization has laid the foundation for future work utilizing Raman spectroscopy to assess therapeutic efficacy and matrix reorganization imparted by nanoceria. Specifically, this will allow us to better understand the role of HIF1a in matrix reorganization of the tumor microenvironment. By studying the relationship between substrate modulus and nanoceria’s ability to inhibit an ECM that is conducive to tumor formation, we endeavor to show that nanoceria may prevent or even revert tumor conducive microenvironments.

Blood vessel dynamics has been a significant subject in cardiology and internal medicine, and pulse wave velocity (PWV) on artery vessels is a classic evaluation of arterial distensibility, and has never been ascertained as a cardiovascular risk marker. The aim of this study is to develop a high speed imaging technique to capture the pulsatile motion on mouse retina arteries with the ability to quantify PWV on any arterial vessels. We demonstrate a new non-invasive method to assess the vessel dynamics on mouse retina. A Swept-source optical coherence tomography (SS-OCT) system is used for imaging micro-scale blood vessel motion. The phase-stabilized SS-OCT provides a typical displacement sensitivity of 20 nm. The frame rate of imaging is ~16 kHz, at A-line rate of ~1.62 MHz, which allows the detection of transient pulse waves with adequate temporal resolution. Imaging volumes with repeated B-scans are obtained on mouse retina capillary bed, and the mouse oxymeter signal is recorded simultaneously. The pulse wave on artery and vein are resolved, and with the synchronized heart beat signal, the temporal delay on different vessel locations is determined. The vessel specific measurement of PWV is achieved for the first time with SS-OCT, for pulse waves propagating more than 100 cm/s. Using the novel methodology of retinal PWV assessment, it is hoped that the clinical OCT scans can provide extended diagnostic information of cardiology functionalities.

Minimally invasive operations require surgeons to make difficult cuts to blood vessels and other tissues with impaired tactile and visual feedback. This leads to inadvertent cuts to blood vessels hidden beneath tissue, causing serious health risks to patients and a non-reimbursable financial burden to hospitals. Intraoperative imaging technologies have been developed, but these expensive systems can be cumbersome and provide only a high-level view of blood vessel networks. In this research, we propose a lean reflectance-based system, comprised of a dual wavelength LED, photodiode, and novel signal processing algorithms for rapid vessel characterization. Since this system takes advantage of the inherent pulsatile light absorption characteristics of blood vessels, no contrast agent is required for its ability to detect the presence of a blood vessel buried deep inside any tissue type (up to a cm) in real time. Once a vessel is detected, the system is able to estimate the distance of the vessel from the probe and the diameter size of the vessel (with a resolution of ~2mm), as well as delineate the type of tissue surrounding the vessel. The system is low-cost, functions in real-time, and could be mounted on already existing surgical tools, such as Kittner dissectors or laparoscopic suction irrigation cannulae. Having been successfully validated ex vivo, this technology will next be tested in a live porcine study and eventually in clinical trials.

Three-dimensional tissue culture presents a more biologically relevant environment in which to perform drug development than conventional two-dimensional cell culture. However, obtaining high-content information from inside three dimensional tissue has presented an obstacle to rapid adoption of 3D tissue culture for pharmaceutical applications. Biodynamic imaging is a high-content three-dimensional optical imaging technology based on low-coherence interferometry and digital holography that uses intracellular dynamics as high-content image contrast. In this paper, we use biodynamic imaging to compare pharmaceutical responses to Taxol of three-dimensional multicellular spheroids grown by three different growth techniques: rotating bioreactor, hanging-drop and plate-grown spheroids. The three growth techniques have systematic variations among tissue cohesiveness and intracellular activity and consequently display different pharmacodynamics under identical drug dose conditions. The in vitro tissue cultures are also compared to ex vivo living biopsies. These results demonstrate that three-dimensional tissue cultures are not equivalent, and that drug-response studies must take into account the growth method.

Cells shape or density is an important marker of tissues pathology. However, individual cells are difficult to observe in thick tissues frequently presenting highly scattering structures such as collagen fibers. Endogenous techniques struggle to image cells in these conditions. Moreover, exogenous contrast agents like dyes, fluorophores or nanoparticles cannot always be used, especially if non-invasive imaging is required. Scatterers motion happening down to the millisecond scale, much faster than the fix and highly scattering structures (global motion of the tissue), allowed us to develop a new approach based on the time dependence of the FF-OCT signals. This method reveals hidden cells after a spatiotemporal analysis based on singular value decomposition and wavelet analysis concepts. It does also give us access to local dynamics of imaged scatterers. This dynamic information is linked with the local metabolic activity that drives these scatterers. Our technique can explore subcellular scales with micrometric resolution and dynamics ranging from the millisecond to seconds. By this mean we studied a wide range of tissues, animal and human in both normal and pathological conditions (cancer, ischemia, osmotic shock…) in different organs such as liver, kidney, and brain among others. Different cells, undetectable with FF-OCT, were identified (erythrocytes, hepatocytes…). Different scatterer clusters express different characteristic times and thus can be related to different mechanisms that we identify with metabolic functions. We are confident that the D-FFOCT, by accessing to a new spatiotemporal metabolic contrast, will be a leading technique on tissue imaging and could lead to better medical diagnosis.

The optical opacity of biological tissue has long been a challenge in biomedical optics due to the strong scattering nature of tissue in the optical regime. While most conventional optical techniques attempt to gate out multiply scattered light and use only unscattered light, new approaches in the field of wavefront shaping exploit the time reversible symmetry of optical scattering in order to focus light inside or through scattering media. While these approaches have been demonstrated effectively on static samples, it has proven difficult to apply them to dynamic biological samples since even small changes in the relative positions of the scatterers within will cause the time symmetry that wavefront shaping relies upon to decorrelate. In this paper we investigate the decorrelation curves of acute rat brain slices for thicknesses in the range 1-3 mm (1/e decorrelation time on the order of seconds) using multi-speckle diffusing wave spectroscopy (MSDWS) and compare the results with theoretical predictions. The results of this study demonstrate that the 1/L^2 relationship between decorrelation time and thickness predicted by diffusing wave spectroscopy provides a good rule of thumb for estimating how the decorrelation of a sample will change with increasing thickness. Understanding this relationship will provide insight to guide the future development of biophotonic wavefront shaping tools by giving an estimate of how fast wavefront shaping systems need to operate to overcome the dynamic nature of biological samples.

In this study, we demonstrate that OCT speckle decorrelation techniques can be used to probe intracellular motion in cancer cells. Spheroids and cell pellets were used as a model to probe intracellular motion. ZnCl₂ was used to inhibit mitochondrial motion within the cells. The results reveal the changes in intracellular motion during the spheroid growth phase. Moreover, to modify the motion of mitochondria, cell pellet were exposed to ZnCl₂, and agent known to o impair cellular energy production through inhibition of mitochondrial function. The speckle decorrelation time during the growth phase of spheroids decreased by 35 ms over 21 days and 25 ms during inhibition of mitochondrial motion 10 minutes after exposure to ZnCl₂.

Polarization microscope is a useful technique to observe the optical anisotropic nature of biomedical specimens and provide more microstructural information than the conventional microscope. In this paper, we present a division of focal plane (DoFP) polarimeter based polarization microscope which is capable of imaging both the Stokes vector and the 3×4 Mueller matrix. The Mueller matrix measurement can help us completely understand the polarization properties of the sample and the Stokes vector measurement is a simultaneous technology. First, we calibrate a DoFP polarimeter using the polarization data reduction method for accurate Stokes vector measurements. Second, as the Stokes vector computation for all pixels using the calibrated instrument matrix is usually time consuming, we develop a GPU acceleration algorithm for real time Stokes vector calculations. Third, based on the accurate and fast Stokes vector calculation, we present an optimal 4-states of polarization (4-SoP) illumination scheme for Mueller matrix measurement using the DoFP polarimeter. Finally, we demonstrate the biomedical applications of the DoFP polarimeter based polarization microscope. Experiment results show that the characteristic features of many biomedical samples can be observed in the “polarization staining” images using the circularly polarized light as illumination. In this way, combined with GPU acceleration algorithm, the DoFP polarization microscope has the capacity for real time polarization monitoring of dynamic processes in biological samples.

Background: Deep tissue ablation is the next challenge in ultrafast laser microsurgery. By focusing ultrafast pulses below the tissue surface one can create an ablation void confined to the focal volume. However, as the ablation depth increases in a scattering tissue, increase in the required power can trigger undesired nonlinear phenomena out of focus that restricts our ability to ablate beyond a maximum ablation depth of few scattering lengths. Optical clearing (OC) might reduce the intensity and increase the maximal ablation depth by lowering the refractive index mismatch, and therefore reducing scattering. Some efforts to ablate deeper showed out of focus damage, while others used brutal mechanical methods for clearing. Our clinical goal is to create voids in the scarred vocal folds and inject a biomaterial to bring back the tissue elasticity and restore phonation.
Materials and methods: Fresh porcine vocal folds were excised and applied a biocompatible OC agent (75% glycerol). Collimated transmittance was monitored. The tissue was optically cleared and put under the microscope for ablation threshold measurements at different depths.
Results: The time after which the tissue was optically cleared was roughly two hours. Fitting the threshold measurements to an exponential decay graph indicated that the scattering length of the tissue increased to 83±16 μm, which is more than doubling the known scattering length for normal tissue.
Conclusion: Optical clearing with Glycerol increases the tissue scattering length and therefore reduces the energy for ablation and increases the maximal ablation depth. This technique can potentially improve clinical microsurgery.

In this report we studied human brain activity in the case of bistable visual perception. We proposed a new approach for quantitative characterization of this activity based on analysis of EEG oscillatory patterns and evoked potentials. Accordingly to theoretical background, obtained experimental EEG data and results of its analysis we studied a characteristics of brain activity during decision-making. Also we have shown that decisionmaking process has the special patterns on the EEG data.

The method for calculation of zero conditional Lyapunov exponent from time series has been proposed. Such method is shown to define the degree of synchronization of the regime realized in the system. It has been applied to real experimental neurophysiological time series represented by electroencephalograms of WAG/Rij rats having genetic predisposition to absence-epilepsy. The degree of synchronization in epileptic brain has been found.

The competition of homophily and homeostasis mechanisms taking place in the multilayer network where several layers of connection topologies are simultaneously present as well as the interaction between layers is considered. We have shown that the competition of homophily and homeostasis leads in such networks to the formation of synchronous patterns within the different layers of the network, which may be both the distinct and identical.

In the present paper the mechanism of the global synchronization onset through the formation of the synchronous clusters in complex networks with different topologies of links (scale-free networks, small-world networks, random networks) is studied. We consider the dependencies of integral characteristics of synchronous dynamics (synchronization measure, number of synchronous clusters, etc) on coupling strength between nodes. As a basic element of the node oscillator we consider Kuramoto phase oscillator.

We investigate effects of a linear resonator on spatial electron dynamics in semiconductor superlattice. We have shown that coupling the external resonant system to superlattice leads to occurrence of the additional area of negative differential conductance on the current-voltage characteristic, which does not occur in autonomous system. Furthermore, this region shows great increase of generation frequency, that contains practical interest.

Usually, diffused reflectance or diffused transmittance has been collected respectively when performing the near-infrared spectroscopic measurements. However, glucose-relative spectral signal is quite weak due to the noises from the measuring system and the environment. Previous Monte-Carlo simulation results from our group showed that the spectral magnitude of both diffused reflectance and diffused transmittance can reach the same order. In this talk, it is our aim to further investigate the synergistic effect of diffused reflectance and diffused transmittance for Near Infrared spectral measurements. The diffused reflectance spectra and diffused transmittance spectra of human’s earlobe have been obtained simultaneously by home-made optical probes within the wavelength of 1100-1400nm. Two processing methods---Superposition Method and Division Method are introduced to demonstrate the synergistic effect of reflectance and transmittance. Both of the processing methods are performed on diffused reflectance and diffused transmittance in accordance with corresponding wavelengths. The results show that the combination of diffused reflectance and transmittance can effectively enhances the SNR by reducing the interference caused by individual differences and measuring environmental factors. Moreover, comparatively, the Division Method has a more distinguished effect.

Previous results revealed that the influences, caused by both individual differences an1d measuring environmental factors, would be reduced by using the floating-reference theory (FRT) for diffused reflectance spectra (DRS) -based blood glucose measurement by Near Infrared Spectroscopy (NIRS). The obtained signals could highlight the variation in light intensity which was brought only by the change of glucose concentration. The existing studies on FRT have mainly focused on the diffused reflectance spectra, but rarely involved the diffused transmittance spectra. In this talk, it is our aim to investigate the availability of FRT on the diffused transmittance spectra (DTS) on the basis of Monte Carlo (MC) simulation method. The MC simulations of DTS have been carried out with different glucose concentrations and skin tissue thicknesses. The simulation results show that the floating reference position point of DTS will disappear when the tissue thickness is greater than a certain value. Therefore, the FRT might be applied on thin tissue model for DTS by NIRS.

In vivo cortex optical imaging is of great important for revealing both structural and functional architecture of brain with high temporal-spatial resolution. To reduce the limitation of turbid skull, researchers had to establish various skull windows or directly expose cortex through craniotomy. Here we developed a skull optical clearing method to make skull transparent. Laser speckle contrast imaging technique was used to monitor the cortical blood flow after topical treatment with the optical clearing agents. The results indicated that the image contrast increased gradually, and then maintained at a high level after 15 min for adult mice, which made the image quality and resolution of micro-vessels nearly approximate to those of exposed cortex. Both the cortical blood flow velocity almost kept constant after skull became transparent. Besides, the treatment of physiological saline on the skull could make skull return to the initial state again and the skull could become transparent again when SOCS retreated it. Thus, we could conclude that the skull optical clearing method was rapid, valid, reversible and safe, which provided us available approach for performing the cortical structural and functional imaging at high temporal-spatial resolution.

In this paper we address the problem of revealing and recognition transitions between distinct physiological states using quite short fragments of experimental recordings. With the wavelet-based multifractal analysis we characterize changes of complexity and correlation properties in the stress-induced dynamics of arterial blood pressure in rats. We propose an approach for association revealed changes with distinct physiological regulatory mechanisms and for quantifying the influence of each mechanism.

Speech signal processing is widely used to reduce noise impact in acquired data. During the last decades, wavelet-based filtering techniques are often applied in communication systems due to their advantages in signal denoising as compared with Fourier-based methods. In this study we consider applications of a 1-D double density complex wavelet transform (1D-DDCWT) and compare the results with the standard 1-D discrete wavelet-transform (1DDWT). The performances of the considered techniques are compared using the mean opinion score (MOS) being the primary metric for the quality of the processed signals. A two-dimensional extension of this approach can be used for effective image denoising.

We propose a random perturbation model to describe the variation of laser speckle patterns caused by moving particles in capillary vessels. When passing through probing volume, moving particles encode random perturbations into observed laser speckle patterns. We extract the perturbation envelopes of time-varying laser speckles for tracking the motion of single particle. And, the full-field transverse velocities of flowing particles are obtained by using cross-correlation between the perturbation envelopes. The proposed method is experimentally verified by the use of polymer-microsphere suspension in a glass capillary.

Eye diseases such as macular degeneration and glaucoma have profound consequences on the quality of human life. Without treatment, these diseases can lead to loss of sight. To develop better treatments for retinal diseases, including cell therapies and drug intervention, establishment of an efficient and reproducible 3D native retinal tissue system, enabled over a prolonged culture duration, will be valuable. The retina is a complex tissue, consisting of ten layers with a different density and cellular composition to each. Uniquely, as a light transmitting tissue, retinal refraction of light differs among the layers, forming a good basis to use optical coherence tomography (OCT) in assessing the layered structure of the retina and its change during the culture and treatments. In this study, we develop a new methodology to generate retinal organotypic tissues and compare two substrates: filter paper and collagen hydrogel, to culture the organotypic tissue. Freshly slaughtered pig eyes have been obtained for use in this study. The layered morphology of intact organotypic retinal tissue cultured on two different substrates has been examined by spectral domain OCT. The viability of the tissues has been examined by live/dead fluorescence dye kit to cross validate the OCT images. For the first time, it is demonstrated that the use of a collagen hydrogel supports the viability of retinal organotypic tissue, capable of prolonged culture up to 2 weeks. OCT is a convenient tool for appraising the quality and application of organotypic retinal samples and is important in the development of current organotypic models.

Activity of brain neurons will lead to changes in local blood flow rate (BFR). Thus, it is important to measure the local BFR of cerebral cortex on research of neuron activity in vivo, such as rehabilitation evaluation after stroke, etc. Currently, laser Doppler flowmetry is commonly used for blood flow measurement, however, relatively low resolution limits its application. Optical coherence tomography (OCT) is a powerful noninvasive 3D imaging modality with high temporal and spatial resolutions. Furthermore, OCT can provide flow distribution image by calculating Doppler frequency shift which makes it possible for blood flow rate measurement. In this paper, we applied OCT to measure the blood flow rate of the primary motor cortex in rats. The animal was immobilized and anesthetized with isoflurane, an incision was made along the sagittal suture, and bone was exposed. A skull window was opened on the primary motor cortex. Then, blood flow rate changes in the primary motor cortex were monitored by our homemade spectral domain OCT with a stimulation of the passive movement of the front legs. Finally, we established the relationship between blood flow rate and the test design. The aim is to demonstrate the potential of OCT in the evaluation of cerebral cortex function.

Reactive hyperemia refers to an increase of blood flow in tissue post release of an occlusion in the local vasculature. Measuring the temporal response of reactive hyperemia, post-occlusion in patients has the potential to shed information about microvascular diseases such as systemic sclerosis and diabetes. Laser speckle contrast imaging (LSCI) is an imaging technique capable of sensing superficial blood flow in tissue which can be used to quantitatively assess reactive hyperemia. Here, we employ LSCI using coherent sources in the blue, green and red wavelengths to evaluate reactive hyperemia in healthy human volunteers. Blood flow in the forearms of subjects were measured using LSCI to assess the time-course of reactive hyperemia that was triggered by a pressure cuff applied to the biceps of the subjects. Raw speckle images were acquired and processed to yield blood-flow parameters from a region of interest before, during and after application of occlusion. Reactive hyperemia was quantified via two measures - (1) by calculating the difference between the peak LSCI flow during the hyperemia and baseline flow, and (2) by measuring the amount of time that elapsed between the release of the occlusion and peak flow. These measurements were acquired in three healthy human participants, under the three laser wavelengths employed. The studies shed light on the utility of in vivo LSCI-based flow sensing for non-invasive assessment of reactive hyperemia responses and how they varied with the choice source wavelength influences the measured parameters.