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Valery V. Tuchin,1,2 Kirill V. Larin,3 Martin J. Leahy,4 Ruikang K. Wang5
1N.G. Chernyshevsky Saratov State Univ. (Russian Federation) 2Univ. of Oulu (Finland) 3Univ. of Houston (United States) 4National Univ. of Ireland, Galway (Ireland) 5Univ. of Washington (United States)
This PDF file contains the front matter associated with SPIE Proceedings Volume 9322, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and Conference Committee listing.
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Laser speckle contrast imaging (LSCI) is a non- or minimally- invasive modality for observing relative blood flow or perfusion. Recently, there has been an effort to use LSCI for truly quantitative blood flow measurements. However, this effort has been hampered not only by real theoretical issues, but also by challenges associated with numerous experimental parameters that can potentially impact the calculated contrast values. In this work, we present our efforts at using a nematic liquid crystal, phase-only, spatial light modulator (SLM) to mimic LSCI experiments with precisely controlled experimental parameters. This approach permits the rapid experimental investigation of numerous factors including: The effects of different flow models on LSCI contrast values; the effects of multiple decorrelation times in the same depth of field; the effects of ‘static’ scatterers; and the effects of camera settings relative to speckle decorrelation times, just to name a few. We have found that an SLM is a useful tool for the experimental investigation of LSCI that eliminates many of the experimental variables associated with typical flow model experiments or in vivo experimentation. LSCI experiments with SLMs are a useful intermediary between computer simulations and physical flow models.
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We present system for long-term continuous PPG monitoring, and physical model for PPG analysis. The system is based on ideology of light scattering modulated by the process of RBC aggregation. OXIRATE’s system works in reflection geometry. The sensor is tiny, completely mobile phone compatible, it can be placed nearly everywhere on the body surface. These technical features allow all-night comfortable PPG monitoring that was performed and analyzed. We can define various sleep stages on the basis of different reproducible time-behavior of PPG signal. Our system of PPG monitoring was used also for reflection pulse oximetry and for extreme PPG studies, such as diving.
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We report the first study on using optical coherence elastography (OCE) to quantitatively monitor the elasticity change of the hyaline cartilage during the optical clearing administrated by glucose solution. The measurement of the elasticity is verified using uniaxial compression test, demonstrating the feasibility of using OCE to quantify the Young’s modulus of the cartilage tissue. As the results, we found that the stiffness of the hyaline cartilage increases during the optical clearing of the tissue. This study might be potentially useful for the early detection of osteoarthritis disease.
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Taking accurate measurements of the state of polarization (SOP) is the key for the success of polarization sensitive techniques which can provide rich information on the microstructure of complex scattering media, such as biological tissues. For static or slow varying samples, SOP measurements can be achieved by time-sequential recoding of different polarization components controlled by rotating polarizers and wave plates or temporal modulation devices such as photoelastic modulators or liquid crystal variable retarders. When the sample is moving or changing its status quickly, polarization components recoded at different time may correspond to different SOPs, which can lead to significant errors in the final results. Simultaneous polarization measurements are necessary for probing such dynamic samples. In this paper, using the simultaneously recorded polarization components, we are able to mimic time sequential polarization schemes and evaluate the errors. The results show that the kinetics of the sample will affect the systematic error and an increase in the statistical errors of the measured degree of polarization (DOP). We change the kinetics of samples with different stirring speed, which is indicated by the characteristic time of the auto-correlation function. It is also demonstrated that the simultaneously recorded polarization components reveals additional information on the orientation of fibrous scatterers as well as their translation and rotation kinetics.
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Generation of functional tissue in vitro through tissue engineering technique is a promising direction to repair and replace malfunctioned organ and tissue in the modern medicine for various diseases which could not been treated well by conventional therapy. Similar to the embryo development, the generation of tissue in vitro is a highly dynamic processing. Obtaining the feedback of the processing real time is highly demanded. In this study, a new methodology has been explored aiming to monitor the morphological and mechanical property alteration of bone tissue engineering constructs simultaneously. Optical coherence elastography (OCE) equipped with a LDS V201 permanent magnet shaker and a modulated acoustic radiation force (ARF) to provide a vibration signal, has been used for the real time and non-destructive monitoring. A phantom construct system has been used to optimize the measurement conditions in which agar hydrogel with concentration from 0, 0.75 to 2% with/without hydroxyappatite particles have been injected to 3D porous poly (lactic acid) scaffolds to simulate the collagenous extracellular matrix (ECM) and mineralized ECM. The structural and elastography images of the constructs have clearly demonstrated the linear relation with the increased mechanical property versus the increase of agar concentration within the pores of the scaffolds. The MG63 bone cells seeded in the scaffolds and cultured for 4 weeks have been monitored by the established protocol exhibiting the increased mechanical strength in the pore wall where the ECM or mineralized ECM was assumed to be formed in comparison to empty pores. This study confirms that OCE-ARF could become a valuable tool in regenerative medicine to assess the biological events during in vitro culture and conditioning.
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Three-dimensional tissue-engineered models are increasingly recognised as more physiologically-relevant than standard 2D cell culture for pre-clinical drug toxicity testing. However, many types of conventional toxicity assays are incompatible with dense 3D tissues. This study investigated the use of optical coherence phase microscopy (OCPM) as a novel approach to assess cell death in 3D tissue culture. For 3D micro-spheroid formation Human hepatic C3A cells were encapsulated in hyaluronic acid gels and cultured in 100μl MEME/10%FBS in 96-well plates. After spheroid formation the 3D liver constructs were exposed to acetaminophen on culture day 8. Acetaminophen hepatotoxicity in 3D cultures was evaluated using standard biochemical assays. An inverted OCPM in common path configuration was developed with a Callisto OCT engine (Thorlabs), centred at 930nm and a custom scanning head. Intensity data were used to perform in-depth microstructural imaging. In addition, phase fluctuations were measured by collecting several successive B scans at the same location, and statistics on the first time derivative of the phase, i.e. time fluctuations, were analysed over the acquisition time interval to retrieve overall cell viability. OCPM intensity (cell cluster size) and phase fluctuation statistics were directly compared with biochemical assays. In this study, we investigated optical coherence phase tomography to assess cell death in a 3d liver model after exposure to a prototypical hepatotoxin, acetaminophen. We showed that OCPM has the potential to assess noninvasively and label-free drug toxicity in 3D tissue models.
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Photothermal heating has been an effective mechanism for harvesting light energy by plasmonic resonance. Photothermally generated hyperthermia can alter cell behavior, change cell microenvironment, and promote or suppress cell growth. In the past, colloidal nanoparticles such as gold nanospheres, nanoshells, nanorods, and nanocages have been developed for various applications. Here, we show that nanoporous gold disks (NPGDs) with 400 nm diameter, 75 nm thickness, and 13 nm pores exhibit large specific surface area and effective photothermal light harvesting capability. Another potential application is demonstrated by light-gated, multi-step molecular release of pre-adsorbed R6G fluorescent dye on arrayed NPGDs. Through the use of time-resolved temperature mapping, the spatial and temporal characteristics of photothermal heating in NPGD arrays is successfully demonstrated for both aqueous and air ambient environments. By applying a thermodynamic model to our experimental data, we determined the photothermal conversion efficiency at 56% for NPGD arrays. As a potential application, light-gated, multi-stage release of pre-adsorbed R6G dye molecules from NPGD arrays has been demonstrated. The results establish the foundation that NPGDs can be employed for photothermal light harvesting and light-gated molecular release.
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Photoacoustic (PA) imaging is a non-invasive real-time technique, widely applied to many biomedical imaging studies in the recent years. While most of these studies have been focussed on obtaining an image after reconstruction, various features of time domain signal (e.g. amplitude, width, rise and relaxation time) would provide very high sensitivity in detecting morphological changes in cells during a biological study. Different haematological disorders (e.g., sickle cell anaemia, thalassemia) exhibit significant morphological cellular changes. In this context, this study explores the possibility of utilizing the developed photoacoustic response technique to apply onto blood samples. Results of our preliminary study demonstrate that there is a significant change in signal amplitude due to change in concentration of the blood. Thus it shows the sensitivity of the developed photoacoustic technique towards red blood cell count (related to haematological disease like anaemia). Subsequently, morphological changes in RBC (i.e. swollen and shrunk compared to normal RBC) induced by hypotonic and hypertonic solutions respectively were also experimented. The result shows a distinct change in PA signal amplitude. This would serve as a diagnostic signature for many future studies on cellular morphological disorders.
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Changes in the microcirculation are associated with conditions such as Raynauds disease. Current modalities used to assess the microcirculation such as nailfold capillaroscopy are limited due to their depth ambiguity. A correlation mapping technique was recently developed to extend the capabilities of Optical Coherence Tomography to generate depth resolved images of the microcirculation. Here we present the extension of this technique to microscopy modalities, including confocal microscopy. It is shown that this correlation mapping microscopy technique can extend the capabilities of conventional microscopy to enable mapping of vascular networks in vivo with high spatial resolution.
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Tape stripping on human skin induces mechanical disruptions of the epidermal barrier that lead to minor skin inflammation which leads to temporary changes in microvasculature. On the other hand, when mosquitoes probe the skin for blood feeding, they inject saliva in dermal tissue. Mosquito saliva is known to exert various biological activities, such as dermal mast cell degranulation, leading to fluid extravasation and neutrophil influx. This inflammatory response remain longer than the tape stripping caused inflammation. In this study, we demonstrate the capabilities of swept-source optical coherence tomography (OCT) in detecting in vivo microvascular response of inflammatory human skin. Optical microangiography (OMAG), noninvasive volumetric microvasculature in vivo imaging method, has been used to track the vascular responses after tape stripping and mosquito bite. Vessel density has been quantified and used to correlate with the degree of skin irritation. The proved capability of OMAG technique in visualizing the microvasculature network under inflamed skin condition can play an important role in clinical trials of treatment and diagnosis of inflammatory skin disorders as well as studying mosquito bite’s perception by the immune system and its role in parasite transmission.
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By combining with the phase sensitive optical coherence tomography (PhS-OCT), vibration and surface acoustic wave (SAW) methods have been reported to provide elastography of skin tissue respectively. However, neither of these two methods can provide the elastography in full skin depth in current systems. This paper presents a feasibility study on an optical coherence elastography method which combines both vibration and SAW in order to give the quantitative mechanical properties of skin tissue with full depth range, including epidermis, dermis and subcutaneous fat. Experiments are carried out on layered tissue mimicking phantoms and invivo human forearm and palm skin. A ring actuator generates vibration while a line actuator were used to excited SAWs. A PhS-OCT system is employed to provide the ultrahigh sensitive measurement of the generated waves. The experimental results demonstrate that by the combination of vibration and SAW method the full skin bulk mechanical properties can be quantitatively measured and further the elastography can be obtained with a sensing depth from ~0mm to ~4mm. This method is promising to apply in clinics where the quantitative elasticity of localized skin diseases is needed to aid the diagnosis and treatment.
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Non-invasive measurements of blood velocity at the capillary level are of great interest in many clinical applications. Recently we presented a new method for the quantitative estimation of blood flow velocity, based on the use of the Radon transform. The technique was based on narrow band illumination at 525 nm and tracking of non-uniform distribution of red blood cells within the vessel, rather than the individual blood cells themselves. Here we present a complete error analysis of this useful technique, highlighting the role of camera and illumination choices. Finally we propose in vivo examples on retinal flow imaging and compare the results obtained with this technique to the one proposed by a commercial system.
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Laser Speckle contrast imaging (LSCI) is a non-invasive or minimally invasive method for visualizing blood flow and perfusion in biological tissues. In LSCI the motion of scattering particles results in a reduction in global and regional speckle contrast. A variety of parameters can affect the calculated contrast values in LSCI techniques, including the optical properties of the fluid and surrounding tissue. In typical LSCI where the motion of blood is of interests, optical properties are influenced by hematocrit levels. In this work we considered the combined effects of both the scattering and absorption coefficients on LSCI measurements on a flow phantom. Fluid phantoms consisting of various concentrations of neutrally buoyant ~10 micron microspheres and India ink mixed with DI water were formulated to mimic the optical properties of whole blood with various levels of hematocrit. In these flow studies, it was found that an increase in μa and/or μs led to a decrease in contrast values when all other experimental parameters were held constant. The observed reduction in contrast due to optical property changes could easily be confused with a contrast reduction due to increased flow velocity. These results suggest that optical properties need to be considered when using LSCI to make flow estimates.
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We demonstrate a method for measuring the total velocity components of particle flow using optical coherence tomography. When passing through a probe volume, moving particles cause the intensity variation of backscattered light. The intensity signal contains the velocity information about the particle flow. Such variation is separated into a phase modulation and an amplitude modulation, from which the axial and transverse components of velocity are obtained. The proposed method is experimentally verified using polystyrene particle suspension flow.
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The spectral measurement error is controlled by the resolution and the sensitivity of the spectroscopic instrument and the instability of involved environment. In this talk, the spectral measurement error has been analyzed quantitatively by using the Monte Carlo (MC) simulation. Take the floating reference point measurement for example, unavoidably there is a deviation between the measuring position and the theoretical position due to various influence factors. In order to determine the error caused by the positioning accuracy of the measuring device, Monte Carlo simulation has been carried out at the wavelength of 1310nm, simulating Intralipid solution of 2%. MC simulation was performed with the number of 1010 photons and the sampling interval of the ring at 1μm. The data from MC simulation will be analyzed on the basis ofthinning and calculating method (TCM) proposed in this talk. The results indicate that TCM could be used to quantitatively analyze the spectral measurement error brought by the positioning inaccuracy.
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The current ear feature matching and recognition system, based on Scale Invariant Feature Transform (SIFT) image matching algorithm, can realize the human ear feature matching and detect the displacement of the human ear so as to reproduce the human ear position and posture. However, due to the influence of image acquisition equipment performance and lighting conditions, too dark or too bright background could bring the locally underexposed or overexposed image. This could result in the loss of some image details so as to make it impossible to identity the image and the recognition rate would be reduced. In this talk, the application of image gray level normalization processing can reduce the sensitivity of imaging to light intensity. Accordingly, it will greatly improve the recognition rate of human ears. Furthermore, it has been found that even if the object is stationary, the image matching results still have certain fluctuation changes, which could be caused by the system error. In order to reduce the error, the Background-based Compensation Model (BCM) has been established based on the investigation of the system error brought by the working environment changes. The results show that, BCM can be used to compensate the system errors of ear recognition matching and further improve the matching accuracy of human ear.
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We study effects of the external tilted magnetic field on the generation of sub-THz/THz oscillations in the semiconductor superlattice. We show that this field provides the increased power of harmonics in the THz range. Changing the tilt angle essentially influences the distribution of spectral power of current oscillations in the semiconductor superlattice.
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In the given paper, a relation between time-frequency characteristics of sleep spindles and the age-dependent epileptic activity in WAG/Rij rats is discussed. Analysis of sleep spindles based on the continuous wavelet transform is performed for rats of different ages. It is shown that the epileptic activity affects the time-frequency intrinsic dynamics of sleep spindles.
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In this paper we study mechanisms of the phase synchronization in a model network of Van der Pol oscillators and in the neural network of the brain by consideration of macroscopic parameters of these networks. As the macroscopic characteristics of the model network we consider a summary signal produced by oscillators. Similar to the model simulations, we study EEG signals reflecting the macroscopic dynamics of neural network. We show that the appearance of the phase synchronization leads to an increased peak in the wavelet spectrum related to the dynamics of synchronized oscillators. The observed correlation between the phase relations of individual elements and the macroscopic characteristics of the whole network provides a way to detect phase synchronization in the neural networks in the cases of normal and pathological activity.
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We investigate the stress-induced development of the intracranial hemorrhage in newborn mice with the main attention to its latent stage. Our study is based on the laser speckle contrast imaging of the cerebral venous blood flow and the wavelet-based analysis of experimental data. We study responses of the sagittal sinus in different frequency ranges associated with distinct regulatory mechanisms and discuss significant changes of the spectral power in the frequency area associated with the NO-related endothelial function.
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The improvement of methods for optical clearing agent prediction exerts an important impact on tissue optical clearing technique. The molecular dynamic simulation is one of the most convincing and simplest approaches to predict the optical clearing potential of agents by analyzing the hydrogen bonds, hydrogen bridges and hydrogen bridges type forming between agents and collagen. However, the above analysis methods still suffer from some problem such as analysis of cyclic molecule by reason of molecular conformation. In this study, a molecular effective coverage surface area based on the molecular dynamic simulation was proposed to predict the potential of optical clearing agents. Several typical cyclic molecules, fructose, glucose and chain molecules, sorbitol, xylitol were analyzed by calculating their molecular effective coverage surface area, hydrogen bonds, hydrogen bridges and hydrogen bridges type, respectively. In order to verify this analysis methods, invitro skin samples optical clearing efficacy were measured after 25 min immersing in the solutions, fructose, glucose, sorbitol and xylitol at concentration of 3.5 M using 1951 USAF resolution test target. The experimental results show accordance with prediction of molecular effective coverage surface area. Further to compare molecular effective coverage surface area with other parameters, it can show that molecular effective coverage surface area has a better performance in predicting OCP of agents.
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Hair follicles enjoy continual cycle of anagen, catagen and telogen all life. They not only provide a unique opportunity to study the physiological mechanism of organ regeneration, but also benefit to guide the treatment of organ repair in regenerative medicine. Usually, the histological examination as a gold standard has been applied to determine the stage of hair follicle cycle, but noninvasive classification of hair cycle in vivo remains unsolved. In this study, the thermal infrared imager was applied to measure the temperature change of mouse dorsal skin with hair follicle cycle, and the change of diffuse reflectance was monitored by the optical fiber spectrometer. Histological examination was used to verify the hair follicle stages. The results indicated that the skin temperature increased at the beginning of anagen. After having stayed a high value for several days, the temperature began to decrease. At the same time, the skin diffuse reflectance decreased until the end of this period. Then the temperature increased gradually after slightly decreased when the hair follicle entered into catagen stage, and the diffuse reflectance increased at this time. In telogen, both the temperature and the diffuse reflectance went back to a steady state all the time. Sub-stages of hair follicle cycle could be distinguished based on the joint curves. This study provided a new method to noninvasively recognize the hair follicle stage, and should be valuable for the basic and therapeutic investigations on hair regeneration.
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In ophthalmology, a reliable means of diagnosing glaucoma in its early stages is still an open issue. Past efforts, including forays into fluorescent angiography (FA) and early optical coherence tomography (OCT) systems, to develop a potential biomarker for the disease have been explored. However, this development has been hindered by the inability of the current techniques to provide useful depth and microvasculature information of the optic nerve head (ONH), which have been debated as possible hallmarks of glaucoma progression. We reasoned that a system incorporating a spectral-domain OCT (SD-OCT) based Optical Microangiography (OMAG) system, could allow an effective, non-invasive methodology to evaluate effects on microvasculature by glaucoma. SD-OCT follows the principle of light reflection and interference to produce detailed cross-sectional and 3D images of the eye. OMAG produces imaging contrasts via endogenous light scattering from moving particles, allowing for 3D image productions of dynamic blood perfusion at capillary-level resolution. The purpose of this study was to investigate the optic cup perfusion (flow) differences in glaucomatous and normal eyes. Images from three normal and five glaucomatous subjects were analyzed our OCT based OMAG system for blood perfusion and structural images, allowing for comparisons. Preliminary results from blood flow analysis revealed reduced blood perfusion within the whole-depth region encompassing the Lamina Cribrosa in glaucomatous cases as compared to normal ones. We conclude that our OCT-OMAG system may provide promise and viability for glaucoma screening.
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