This PDF file contains the front matter associated with SPIE Proceedings Volume 10501, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

Numerous potential biomedical sensing applications of surface-enhanced Raman scattering (SERS) have been reported, but its practical use has been limited by the lack of a robust sensing platform. Optical fiber SERS probes show great promise, but are limited by the prominent silica Raman background, which requires the use of bulky optics for filtering the signal collection and excitation delivery paths. In the present study, a SERS microprobe has been designed and developed to eliminate the bottlenecks outlined above. For efficient excitation and delivery of the SERS signal, both hollow core photonic crystal fiber and double clad fiber have been investigated. While the hollow core fiber was still found to have excessive silica background, the double clad fiber allows efficient signal collection via the multi-mode inner cladding. A micro filtering mechanism has been designed, which can be integrated into the tip of the optical fiber SERS probe, providing filtering to suppress silica Raman background and thus avoiding the need for bulky optics. The design also assists in the efficient collection of SERS signal from the sample by rejecting Rayleigh scattered light from the sample. Optical fiber cleaving using ultra-short laser pulses was tested for improved control of the fiber tip geometry. With this miniaturized and integrated filtering mechanism, it is expected that the developed probe will promote the use of SERS for minimally invasive biomedical monitoring and sensing applications in future. The probe could potentially be placed inside a small gauge hypodermic needle and would be compatible with handheld portable spectrometers.

Metal nanoparticles conjugated with DNA oligomers have been intensively studied for a variety of applications, including optical diagnostics. Assays based on aggregation of DNA-coated particles in proportion to the concentration of target analyte have not been widely adopted for clinical analysis, however, largely due to the nonspecific responses observed in complex biofluids. While sample pre-preparation such as dialysis is helpful to enable selective sensing, here we sought to prove that assay encapsulation in hollow microcapsules could remove this requirement and thereby facilitate more rapid analysis on complex samples. Gold nanoparticle-based assays were incorporated into capsules comprising polyelectrolyte multilayer (PEMs), and the response to small molecule targets and larger proteins were compared. Gold nanoparticles were able to selectively sense small Raman dyes (Rhodamine 6G) in the presence of large protein molecules (BSA) when encapsulated. A ratiometric based microRNA-17 sensing assay exhibited drastic reduction in response after encapsulation, with statistically-significant relative Raman intensity changes only at a microRNA-17 concentration of 10 nM compared to a range of 0-500 nM for the corresponding solution-phase response.

It is estimated that the number of new cases of oral cancers worldwide is 529,000 and more than 300,000 deaths each year. The five-year survival rate remains about 50%, and the low survival rate is believed to be due to delayed detection. The primary detection method is through a comprehensive clinical examination by a dentist followed by a biopsy of suspicious lesions. Systematic review and meta-analysis have revealed that clinical examination alone may not be sufficient to cause the clinician to perform a biopsy or refer for biopsy for early detection of OSCC. Therefore, a non-invasive, point-of-Care (POC) detection with high sensitivity and specificity for early detection would be urgently needed, and using salivary biomarkers would be an ideal technology for it. S100 calcium binding protein P (S100P) mRNA presenting in saliva is a potential biomarker for detection of oral cancer. Further, surface enhanced Raman spectroscopy (SERS) has been shown to be a promising POC diagnostic technique. In this research, a SERS-based assay using oligonucleotide strains was developed for the sensitive and rapid detection of S100P. Gold nanoparticles (AuNPs) as a SERS substrate were used for the conjugation with one of two unique 24 base pair oligonucleotides, referred to as left and right DNA probes. A Raman reporter molecule, malachite green isothiocyanate (MGITC), was bound to left-probe-conjugated AuNPs. UV-vis spectroscopy was employed to monitor the conjugation of DNA probes to AuNPs. The hybridization of S100P target to DNA-conjugated AuNPs in sandwich-assay format was confirmed by Raman spectroscopy and shown to yield and R² of 0.917 across the range of 0-200 nM and a limit of detection of 3 nM.

Research toward development of point-of-care (POC) technologies is emerging as a means for diagnosis and monitoring of patients outside the hospital. These POC devices typically utilize assays capable of detecting low level biomarkers indicative of specific diseases. L-citrulline, an α-amino acid produced in the intestinal mucosa cells, is one such biomarker typically found circulating within the plasma at physiological concentrations of ~40 μM. Researchers have found that intestinal enterocyte malfunction causes its level to be significantly lowered, establishing it as a potential diagnostic biomarker for gut function. Our research group has proposed the development of a surface enhanced Raman spectroscopy (SERS) based assay, using vertical flow paper fluidics, for citrulline detection. The assay consists of a fluorescently active, Raman reporter labeled aptamer conjugated on gold nanoparticles. The aptamer changes its confirmation on binding to its target, which in turn changes the distance between the Raman active molecule and the nanoparticle surface. These particles were embedded within a portable chip consisting of cellulose-based paper. After the chips were loaded with different concentrations of free L-citrulline in phosphate buffer, time was given for the assay to interact with the sample. A handheld Raman spectrometer (638 nm; Ocean Optics) was used to measure the SERS intensity. Results showed decrease in intensity with increasing concentration of L-citrulline (0-50μM).

Central to minimizing the long- and short-term complications associated with diabetes is careful monitoring and maintenance of blood glucose at normal levels. Towards replacing conventionally used finger-prick glucose testing, indwelling continuous glucose monitors (CGMs) based on amperometric electrodes have been introduced to the market. Envisioned to lead to a CGM with an increased lifetime, we report herein a fluorescently-labeled competitive binding assay contained within a hydrogel membrane whose glucose response is measured via a novel portable system. The optical system design included a laser source, bifurcated fiber, laser filter and simple fiber coupled spectrometer to obtain the change in FRET pair ratio of the assay. Glucose response of the assay in free solution was measured using this system across the physiologic range (0-200 mg/dL). The FRET pair ratio signal was seen to increase with glucose and the standard error of calibration was 22.42 mg/dL with a MARD value of 14.85%. When the assay was contained within the hydrogel membrane’s central cavity and similarly analyzed, the standard error increased but the assay maintained its reversibility.

Observing the circular dichroism (CD) caused by organic molecules in biological fluids can provide powerful indicators of patient health and provide diagnostic clues for treatment. Methods for this kind of analysis involve tabletop devices that weigh tens of kilograms with costs on the order of tens of thousands of dollars, making them prohibitive in point-of-care diagnostic applications. In an e ort to reduce the size, cost, and complexity of CD estimation systems for point-of-care diagnostics, we propose a novel method for CD estimation that leverages a vortex half-wave retarder in between two linear polarizers and a two-dimensional photodetector array to provide an overall complexity reduction in the system. This enables the measurement of polarization variations across multiple polarizations after they interact with a biological sample, simultaneously, without the need for mechanical actuation. We further discuss design considerations of this methodology in the context of practical applications to point-of-care diagnostics.

Mid-infrared spectroscopy hyphenated with micro-dialysis is an excellent method for monitoring metabolic blood parameters as it enables the concurrent, reagent-free and precise measurement of multiple clinically relevant substances such as glucose, lactate and urea in micro-dialysates of blood or interstitial fluid. For a marketable implementation, quantum cascade lasers (QCL) seem to represent a favourable technology due to their high degree of miniaturization and potentially low production costs. In this work, an external cavity (EC) - QCL-based spectrometer and two Fourier-transform infrared (FTIR) spectrometers were benchmarked with regard to the precision, accuracy and long-term stability needed for the monitoring of critically ill patients. For the tests, ternary aqueous solutions of glucose, lactate and mannitol (the latter for dialysis recovery determination) were measured in custom-made flow-through transmission cells of different pathlengths and analyzed by Partial Least Squares calibration models. It was revealed, that the wavenumber tuning speed of the QCL had a severe impact on the EC-mirror trajectory due to matching the digital-analog-converter step frequency with the mechanical resonance frequency of the mirror actuation. By selecting an appropriate tuning speed, the mirror oscillations acted as a hardware smoothing filter for the significant intensity variations caused by mode hopping. Besides the tuning speed, the effects of averaging over multiple spectra and software smoothing parameters (Savitzky-Golay-filters and FT-smoothing) were investigated. The final settings led to a performance of the QCL-system, which was comparable with a research FTIR-spectrometer and even surpassed the performance of a small FTIR-mini-spectrometer.

In this study a diffraction based sensor has been developed for bio molecular sensing applications and performing assays in real time. A diffraction grating fabricated on a glass substrate produced diffraction patterns both in transmission and reflection when illuminated by a laser diode. We used zeroth order I_(0,0) as reference and first order I_(0,1) as signal channel and conducted ratiometric measurements that reduced noise by more than 50 times. The ratiometric approach resulted in a very simple instrumentation with very high sensitivity. In the past, we have shown refractive index measurements both for bulk and surface adsorption using the diffractive self-referencing approach. In the current work we extend the same concept to higher diffraction orders. We have considered order I_(0,1) and I_(1,1) and performed ratiometric measurements I_(0,1)/I_(1,1) to eliminate the common mode fluctuations. Since orders I_(0,1) and I_(1,1) behaved opposite to each other, the resulting ratio signal amplitude increased more than twice compared to our previous results. As a proof of concept we used different salt concentrations in DI water. Increased signal amplitude and improved fluid injection system resulted in more than 4 times improvement in detection limit, giving limit of detection 1.3×10^-7 refractive index unit (RIU) compared to our previous results. The improved refractive index sensitivity will help significantly for high sensitivity label free bio sensing application in a very cost-effective and simple experimental set-up.

Lens-free holographic microscopy is a promising diagnostic approach because it is cost-effective, compact, and suitable for point-of-care applications, while providing high resolution together with an ultra-large field-of-view. It has been applied to biomedical sensing, where larger targets like eukaryotic cells, bacteria, or viruses can be directly imaged without labels, and smaller targets like proteins or DNA strands can be detected via scattering labels like micro- or nano-spheres. Automated image processing routines can count objects and infer target concentrations. In these sensing applications, sensitivity and specificity are critically affected by image resolution and signal-to-noise ratio (SNR). Pixel super-resolution approaches have been shown to boost resolution and SNR by synthesizing a high-resolution image from multiple, partially redundant, low-resolution images. However, there are several computational methods that can be used to synthesize the high-resolution image, and previously, it has been unclear which methods work best for the particular case of small-particle sensing. Here, we quantify the SNR achieved in small-particle sensing using regularized gradient-descent optimization method, where the regularization is based on cardinal-neighbor differences, Bayer-pattern noise reduction, or sparsity in the image. In particular, we find that gradient-descent with sparsity-based regularization works best for small-particle sensing. These computational approaches were evaluated on images acquired using a lens-free microscope that we assembled from an off-the-shelf LED array and color image sensor. Compared to other lens-free imaging systems, our hardware integration, calibration, and sample preparation are particularly simple. We believe our results will help to enable the best performance in lens-free holographic sensing.

Despite numerous advances, malaria continues to kill nearly half a million people globally every year. New analytical methods and diagnostics are critical to understanding how treatments under development affect the lifecycle of malaria parasites. A biomarker that has been gaining interest is the "malaria pigment" hemozoin. This byproduct of hemoglobin digestion by the parasite has a unique spectral signature but is difficult to differentiate from hemoglobin and other tissue chromophores. Hemozoin can be detected in blood samples, but only utilizing approaches that require specialized training and facilities.

Diffuse optical spectroscopy (DOS) is a noninvasive sensing technique that is sensitive to near-infrared absorption and scattering and capable of probing centimeter-deep volumes of tissue in vivo. DOS is relatively low-cost, does not require specialized training and thus potentially suitable for use in low-resource settings. In this work, we assess the potential of DOS to detect and quantify the presence of hemozoin noninvasively and at physiologically relevant levels. We suspended synthetic hemozoin in Intralipid-based tissue-simulating phantoms in order to mimic malaria infection in multiply-scattering tissue. Using a fiber probe that combines frequency-domain and continuous-wave broadband DOS (650-1000 nm), we detected hemozoin concentrations below 250 ng/ml, which corresponds to parasitemia sensitivities comparable to modern rapid diagnostic tests. We used the experimental variability to simulate and estimate the sensitivity of DOS to hemozoin in tissue that includes hemoglobin, water, and lipid under various tissue oxygen saturation levels. The results indicate that with increased precision, it may be possible to detect Hz noninvasively with DOS.

In this paper, we find the effective feature values of skin textures captured by non-contact camera to monitor piloerection on the skin for emotion estimation. Recently, emotion estimation is required for service robots to interact with human more naturally. There are a lot of researches of estimating emotion and additional methods are required to improve emotion estimation because using only a few methods may not give enough information for emotion estimation. In the previous study, it is necessary to fix a device on the subject’s arm for detecting piloerection, but the contact monitoring can be stress itself and distract the subject from concentrating in the stimuli and evoking strong emotion. So, we focused on the piloerection as the object obtained with non-contact methods. The piloerection is observed as goose bumps on the skin when the subject is emotionally moved, scared and so on. This phenomenon is caused by contraction of arrector pili muscles with the activation of sympathetic nervous system. This piloerection changes skin texture. Skin texture is important in the cosmetic industry to evaluate skin condition. Therefore, we thought that it will be effective to evaluate the condition of skin texture for emotion estimation. The evaluations were performed by extracting the effective feature values from skin textures captured with a high resolution camera. The effective feature values should have high correlation with the degree of piloerection. In this paper, we found that standard deviation of short-line inclination angles in the texture is well correlated with the degree of piloerection.

The demand for self-measured blood pressure self-monitoring device has much increased due to cardiovascular diseases have become leading causes of death for aging population. Currently, the primary non-invasive blood pressure monitoring method is cuff-based. It is well developed and accurate. However, the measuring process is not comfortable, and it cannot provide a continuous measurement. To overcome this problem, methods such as tonometry, volume clamp method, photoplethysmography, pulse wave velocity, and pulse transit time are reported. However, the limited accuracy hindered its application for diagnostics. To perform sequential blood pressure measurement with a high accuracy and long-term examination, we apply moiré interferometry to measure wrist skin vibration induced by radial artery. To achieve this goal, we developed a miniaturized device that can perform moiré interferometry around the wrist region. The 0.4-mm-pitched binary grating and tattoo sticker with 0.46 mm-pitched stripe pattern are used to perform geometric moiré. We demonstrated that the sensitivity and accuracy of this integrated system were sufficient to monitor arterialinduced skin vibration non-invasively. Our developed system was validated with ECG signals collected by a commercial system. According to our studies from measurement, the repeatability of wrist pulsation measurement was achieved with an accuracy of 99.1% in heart rate. A good repeatability of wrist pulse measurement was achieved. Simulations and experiments are both conducted in this paper and prove of geometrical moiré method a suitable technique for arterial-induced skin vibration monitoring.

Dysphagia (swallowing difficulty) increases risk for malnutrition and affects at least 15% of American older adults, and 590 million people worldwide. Malnutrition is associated with increased mortality, increased morbidity, decreased quality of life, and accounts for over $15 billion (USD) health-care related costs each year. While modified texture diets (e.g., puréed food) reduce the risk of choking, quality assurance is necessary for monitoring nutrient density to ensure food meets nutritional requirements. However, current methods are subjective and time consuming. The purpose of this study was to investigate the feasibility of optical techniques for an objective assessment of food nutrient density in puréed samples. Motivated by a theoretical optical dilution model, broadband spectral images of commercially prepared purée samples were acquired. Specifically, 13 flavors at five dilutions relative to initial concentration, each with six replicates, were acquired for a total of 390 samples. Purée samples were prepared and loaded onto a white reflectance back plane to maximize photon traversal path length through the purée. The sample was illuminated with a tungsten-halogen illumination source fitted with a front glass fabric diffuser for spatially homogeneous illumination. This broadband illuminant was chosen to observe as many food-light spectral absorbance interactions as possible. Flavor-stratified correlation analysis was performed on this food image dataset to investigate the relationship between nutritional information and color space transformations. A special case of blueberry is presented as the effect of anthocyanins was quantitatively observed through normalized spectral trends in response to pH perturbations across dilutions.

We investigated the appropriate hemodynamic parameters of functional near-infrared spectroscopy (fNIRS) to best differentiate the discomfort intensity related to swallowing in healthy subjects wearing different types of simulated dentures. Thirty-one subjects performed 4 sessions of water-swallowing task in a sitting position without or with wearing any one of the 3 simulated dentures under continuous fNIRS monitoring of the prefrontal cortex. The simulated dentures were consisted of a flat maxillary palatal bar and bilateral support attachment to the teeth and gum. The palatal bar positions were arranged along with the anterior, middle, or posterior areas of the upper jaw. The cumulative values of oxyhemoglobin and deoxyhemoglobin concentration changes were calculated for periods of 5 - 10 s from completion of swallowing. Subjective rating of swallowing-discomfort was the lowest without wearing denture and increased along with the palatal bar position placed from anterior to posterior. Cumulated deoxyhemoglobin signals calculated for 10 s post-swallowing event in the left frontopolar prefrontal cortex showed the best correlation with the subjective rating of discomfort in swallowing with dentures. These neurological signatures would be beneficial to detect swallowing discomfort in elderly denture wearers who have difficulty in communicating with dentists.

Introduction: Sudden physical trauma to the spinal cord results in acute spinal cord injury (SCI), leading to spinal cord (SC) tissue destruction, acute inflammation, increased SC intraparenchymal pressure, and tissue ischemia, hypoxia, and cellular necrosis. The ability to monitor SC tissue viability at subcellular level, using a real-time noninvasive method, would be extremely valuable to clinicians for estimating acute SCI damage, and adjusting and monitoring treatment in the intensive care setting. This study examined the feasibility and sensitivity of a custommade near infrared spectroscopy (NIRS) sensor to monitor the oxidation state of SC mitochondrial cytochrome aa3 (CCO), which reflects the subcellular damage of SC tissue in an animal model of SCI.

Methods: Six anesthetized Yorkshire pigs were studied using a custom-made multi-wavelength NIRS system with a miniaturized optical sensor applied directly on the surgically exposed SC at T9. The oxidation states of SC tissue hemoglobin and CCO were monitored before, during and after acute SCI, and during mean arterial pressure alterations.

Results: Non-invasive NIRS monitoring reflected changes in SC tissue CCO, simultaneous but independent of changes in hemoglobin saturation following acute SCI. A consistent decrease in SC tissue CCO chromophore concentration (-1.98 ± 2.1 ab, p<0.05) was observed following SCI, indicating progressive SC cellular damage at the injury site. Elevation of mean arterial pressure can reduce SC tissue damage as suggested by different researchers and observed by significant increase in SC tissue CCO concentration (1.51 ± 1.7 ab, p<0.05) in this study.

Conclusions: This pilot study indicates that a novel miniaturized multi-wave NIRS sensor has the potential to monitor post-SCI changes of SC cytochrome aa3 oxygenation state in real time. Further development of this method may offer new options for improved SCI care.

Background: ‘Tisseel’ (Baxter Healthcare, Deerfield, IL) is a fibrin-based sealant that is commonly used during spine surgery to augment dural repairs. We wish to intra-operatively secure a near infrared spectroscopy (NIRS) sensor to the dura in order to monitor the tissue hemodynamics of the underlying spinal cord. To determine if ‘Tisseel’ sealant adversely attenuates NIR photon transmission. Methods: We investigated ‘Tisseel’ in both an in vitro and in vivo paradigm. For in vitro testing, we used a 1 mm pathlength cuvette containing either air or ‘Tisseel’ interposed between a NIR light source (760 and 850 nm) and a photodiode detector and compared transmittance. For in vivo testing, a continuous wave (760 and 850 nm) spatiallyresolved NIRS device was placed over the triceps muscle using either conventional skin apposition (overlying adhesive bandage) or bioadhesion with ‘Tisseel’. Raw optical data and tissue saturation index (TSI%) collected at rest were compared. Results: In-vitro NIR light absorption by ‘Tisseel’ was very high, with transmittance reduced by 95% compared to air. In-vivo muscle TSI% values were 80% with conventional attachment and 20% using fibrin glue. Conclusion: The optical properties of ‘Tisseel’ significantly attenuate NIR light during in-vitro transmittance and critically compromise photon transmission in-vivo.

Low back pain (LBP) is a complex disease that can be cause by a variety of reasons. Now LBP has become a very common and severe disease among kinds of occupational groups with showing a younger trend. The traditional diagnosis relies on complicated imaging modalities and other dangerous and invasive methods. Noninvasive near-infrared spectroscopy (NIRS) is noninvasive and convenient, and has been successful used in point-of-care diagnosis. Here, we attempt to explore NIRS’s application in in low back pain diagnosis and the effect of aid-use of Chinese cupping procedure. 13 LBP patients and 13 healthy subjects participated in NIRS measurements of concentrations of oxy- and deoxy-hemoglobins (∆[HbO₂] and ∆[Hb]) at the middle of the lumbar spine. It was showed that there was significant differences (p < 0.001) between healthy subjects and LBP patients after cupping procedure, while insignificant before cupping. Moreover, it was found that healthy subjects showed stronger responses to cupping procedure than LBP patients, with prominently higher concentration of ∆[HbO₂] and ∆[Hb]. It indicates the potential of NIRS in noninvasive, measurable and straightforward monitoring/therapeutic effect evaluation of LBP with bedside and point-of-care monitoring capability.

Near-infrared spectroscopy (NIRS) is a noninvasive method for monitoring tissue oxygen saturation (StO₂). Many commercial NIRS devices are presently available. However, the precision of those devices is relatively poor because they are using the reflectance-model with which it is difficult to obtain the blood volume and other unchanged components of the tissue. Human webbing is a thin part of the hand and suitable to measure spectral transmittance. In this paper, we present a method for measuring StO₂ of human webbing from a transmissive continuous-wave nearinfrared spectroscopy (CW-NIRS) data. The method is based on the modified Beer-Lambert law (MBL) and it consists of two steps. In the first step, we give a pressure to the upstream region of the measurement point to perturb the concentration of deoxy- and oxy-hemoglobin as remaining the other components and measure the spectral signals. From the measured data, spectral absorbance due to the components other than hemoglobin is calculated. In the second step, spectral measurement is performed at arbitrary time instance and the spectral absorbance obtained in the step 1 is subtracted from the measured absorbance. The tissue oxygen saturation (StO₂) is estimated from the remained data. The method was evaluated on an arterial occlusion test (AOT) and a venous occlusion test (VOT). In the evaluation experiment, we confirmed that reasonable values of StO₂ were obtained by the proposed method.

Imaging photoplethysmography (iPPG) allows non-contact, concomitant measurement and visualization of peripheral blood flow using just an RGB camera. Most iPPG methods require a window of temporal data and complex computation, this makes real-time measurement and spatial visualization impossible. We present a fast,“window-less”, non-contact imaging photoplethysmography method, based on a tissue-like model of the skin, that allows accurate measurement of heart rate and heart rate variability parameters. The error in heart rate estimates is equivalent to state-of-the-art techniques and computation is much faster.

Non-contact vital-sign estimation allows the monitoring of physiological parameters (such as heart rate, respiratory rate, and peripheral oxygen saturation) without contact electrodes or sensors. Our recent work has demonstrated that a convolutional neural network (CNN) can be used to detect the presence of a patient and segment the patient’s skin area for vital-sign estimation, thus enabling the automatic continuous monitoring of vital signs in a hospital environment.

In a study approved by the local Research Ethical Committee, we made video recordings of pre-term infants nursed in a Neonatal Intensive Care Unit (NICU) at the John Radcliffe Hospital in Oxford, UK. We extended the CNN model to detect the head, torso and diaper of the infants. We extracted multiple photoplethysmographic imaging (PPGi) signals from each body part, analysed their signal quality, and compared them with the PPGi signal derived from the entire skin area. Our results demonstrated the benefits of estimating heart rate combined from multiple regions of interest using data fusion. In the test dataset, we achieved a mean absolute error of 2.4 beats per minute for 80% (31.1 hours) from a total recording time of 38.5 hours for which both reference heart rate and video data were valid.

The inter-beat-interval (time period of the cardiac cycle) changes slightly for every heartbeat; this variation is measured as Heart Rate Variability (HRV). HRV is presumed to occur due to interactions between the parasym- pathetic and sympathetic nervous system. Therefore, it is sometimes used as an indicator of the stress level of an individual. HRV also reveals some clinical information about cardiac health. Currently, HRV is accurately measured using contact devices such as a pulse oximeter. However, recent research in the field of non-contact imaging Photoplethysmography (iPPG) has made vital sign measurements using just the video recording of any exposed skin (such as a person's face) possible. The current signal processing methods for extracting HRV using peak detection perform well for contact-based systems but have poor performance for the iPPG signals. The main reason for this poor performance is the fact that current methods are sensitive to large noise sources which are often present in iPPG data. Further, current methods are not robust to motion artifacts that are common in iPPG systems. We developed a new algorithm, CameraHRV, for robustly extracting HRV even in low SNR such as is common with iPPG recordings. CameraHRV combined spatial combination and frequency demodulation to obtain HRV from the instantaneous frequency of the iPPG signal. CameraHRV outperforms other current methods of HRV estimation. Ground truth data was obtained from FDA-approved pulse oximeter for validation purposes. CameraHRV on iPPG data showed an error of 6 milliseconds for low motion and varying skin tone scenarios. The improvement in error was 14%. In case of high motion scenarios like reading, watching and talking, the error was 10 milliseconds.

Optical monitoring of arterial blood oxygenation, SpO₂, using cameras has recently been shown feasible by measuring the relative amplitudes of the remotely sensed PPG waveforms captured at different wavelengths. SvO₂ measures the venous blood oxygenation which together with SpO₂ provides an indication of tissue oxygen consumption. In contrast to SpO₂ it usually still requires a blood sample from a pulmonary artery catheter. In this work we present a method which suggests simultaneous estimation of SpO₂ and SvO₂ with a camera. Contrary to earlier work, our method does not require external cuffs leading to better usability and improved comfort. Since the arterial blood varies synchronously with the heart rate, all frequencies outside the heart rate band are typically filtered out for SpO₂ measurements. For SvO₂ estimation, we include intensity variations in the respiratory frequency range since respiration modulates venous blood due to intrathoracic pressure variations in the chest and abdomen. Consequently, under static conditions, the two dominant components in the PPG signals are respiration and pulse. By measuring the amplitude ratios of these components, it seems possible to monitor both SpO₂ and SvO₂ continuously. We asked healthy subjects to follow an auditory breathing pattern while recording the face and hand. Results show a difference in estimated SpO₂ and SvO₂ values in the range 5-30 percent for both anatomical locations, which is normal for healthy people. This continuous, non-contact, method shows promise to alert the clinician to a change in patient condition sooner than SpO₂ alone.

Imaging photoplethysmography (iPPG) has attracted much attention over the last years. The vast majority of works focuses on methods to reliably extract the heart rate from videos. Only a few works addressed iPPGs ability to exploit spatio-temporal perfusion pattern to derive further diagnostic statements.

This work directs at the spatio-temporal analysis of blood perfusion from videos. We present a novel algorithm that bases on the two-dimensional representation of the blood pulsation (perfusion map). The basic idea behind the proposed algorithm consists of a pairwise estimation of time delays between photoplethysmographic signals of spatially separated regions. The probabilistic approach yields a parameter denoted as perfusion speed. We compare the perfusion speed versus two parameters, which assess the strength of blood pulsation (perfusion strength and signal to noise ratio).

Preliminary results using video data with different physiological stimuli (cold pressure test, cold face test) show that all measures are influenced by those stimuli (some of them with statistical certainty). The perfusion speed turned out to be more sensitive than the other measures in some cases. However, our results also show that the intraindividual stability and interindividual comparability of all used measures remain critical points.

This work proves the general feasibility of employing the perfusion speed as novel iPPG quantity. Future studies will address open points like the handling of ballistocardiographic effects and will try to deepen the understanding of the predominant physiological mechanisms and their relation to the algorithmic performance.

Imaging photoplethysmography uses camera image sensors to measure variations in light absorption related to the delivery of the blood volume pulse to peripheral tissues. The characteristics of the measured BVP waveform depends on the spectral absorption of various tissue components including melanin, hemoglobin, water, and yellow pigments. Signal quality and artifact rejection can be enhanced by taking into account the spectral properties of the BVP waveform and surrounding tissue. The current literature regarding the spectral relationships of remote PPG is limited. To supplement this fundamental data, we present an analysis of remotely-measured, visible and near-infrared spectroscopy to better understand the spectral signature of remotely measured BVP signals. To do so, spectra were measured from the right cheek of 25, stationary participants whose heads were stabilized by a chinrest. A collimating lens was used to collect reflected light from a region of 3 cm in diameter. The spectrometer provided 3 nm resolution measurements from 500-1000 nm. Measurements were acquired at a rate of 50 complete spectra per second for a period of five minutes. Reference physiology, including electrocardiography was simultaneously and synchronously acquired. The spectral data were analyzed to determine the relationship between light wavelength and the resulting remote-BVP signal-to-noise ratio and to identify those bands best suited for pulse rate measurement. To our knowledge this is the most comprehensive dataset of remotely-measured spectral iPPG data. In due course, we plan to release this dataset for research purposes.

Systolic blood pressure (SBP) is highly sensitive to various factors such as psychological stress, and hence its continuous monitoring is essential to evaluate different health conditions. However, conventional sphygmomanometers cannot continuously measure SBP given the time-consuming setup based on a pressure cuff. Moreover, continuous biological signal monitoring is more comfortable when no sensors are attached. A solution for continuous SBP estimation is based on pulse transit time (PTT), which determines the time difference between two pulse waves at different body parts. In previous studies, we successfully measured the PTT using a contactless setup composed by two digital color cameras recording the face and hand of subjects. Then, the acquired images were transformed into blood volume by combining multiple regression analysis and a Monte Carlo method. As a result, the delay among images allowed to determine the PPT from pulse waves. In this study, we simultaneously measured SBP and PTT by using a sphygmomanometer and the two cameras, respectively. We evaluated SBP increases (i.e., stressful situations) and the corresponding PPT by asking participants to either grasp a handgrip or momentarily interrupting breath. We also determined the SBP and PTT without asking for such exercises. Comparison results show that the mean PTT under stress was significantly lower than that without stress, which is consistent with an increased SBP. Finally, we related the SBP and PTT by a nonlinear formula with a coefficient of determination of 0.59, thus confirming the effectiveness of the proposed system.

Plethysmogram is the periodic variation in blood volume due to the cardiac pulse traveling through the body. Photo-plethysmograph (PPG) has been widely used to assess the cardiovascular system such as heart rate, blood pressure, cardiac output, vascular compliance. We have previously proposed a non-contact PPG imaging method using a digital red-green-blue camera. In the method, the Monte Carlo simulation for light transport is used to specify a relationship among the RGB-values and the concentrations of oxygenated hemoglobin (C_HbO) and deoxygenated hemoglobin (C_HbR). The total hemoglobin concentration (C_HbT) can be calculated as a sum of C_HbO and C_HbR. Applying the fast Fourier transform (FFT) band pass filters to each pixel of the sequential images for C_HbT along the time line, two-dimentional plethysmogram can be reconstructed. In this study, we further extend the method to imaging the arterial oxygen saturation (SaO₂). The PPG signals for both C_HbO and C_HbR are extracted by the FFT band pass filter and the pulse wave amplitudes (PWAs) of C_HbO and C_HbR are calculated. We assume that the PWA for C_HbO and that for CHbR are decreased and increased as SaO₂ is decreased. The ratio of PWA for C_HbO and that for C_HbR are associated to the reference value of SaO₂ measured by a commercially available pulse oximeter, which provide an empirical formula to estimate SaO₂ from the PPG signal at each pixel of RGB image. In vivo animal experiments with rats during varying the fraction of inspired oxygen (FiO₂) demonstrated the feasibility of the proposed method.

Monitoring respiration during neonatal sleep is notoriously difficult due to the nonstationary nature of the signals and the presence of spurious noise. Current approaches rely on the use of adhesive sensors, which can damage the fragile skin of premature infants. Recently, non-contact methods using low-cost RGB cameras have been proposed to acquire this vital sign from (a) motion or (b) photoplethysmographic signals extracted from the video recordings. Recent developments in deep learning have yielded robust methods for subject detection in video data. In the analysis described here, we present a novel technique for combining respiratory information from high-level visual descriptors provided by a multi-task convolutional neural network. Using blind source separation, we find the combination of signals which best suppresses pulse and motion distortions and subsequently use this to extract a respiratory signal. Evaluation results were obtained from recordings on 5 neonatal patients nursed in the Neonatal Intensive Care Unit (NICU) at the John Radcliffe Hospital, Oxford, UK. We compared respiratory rates derived from this fused breathing signal against those measured using the gold standard provided by the attending clinical staff. We show that respiratory rate (RR) be accurately estimated over the entire range of respiratory frequencies.

Obstructive sleep apnea (OSA) affects 20% of the adult population, and is associated with cardiovascular and cognitive morbidities. However, it is estimated that up to 80% of treatable OSA cases remain undiagnosed. Cur- rent methods for diagnosing OSA are expensive, labor-intensive, and involve uncomfortable wearable sensors. This study explored the feasibility of non-contact biophotonic assessment of OSA cardiovascular biomarkers via photoplethysmography imaging (PPGI). In particular, PPGI was used to monitor the hemodynamic response to obstructive respiratory events. Sleep apnea onset was simulated using Muller's maneuver in which breathing was obstructed by a respiratory clamp. A custom PPGI system, coded hemodynamic imaging (CHI), was positioned 1 m above the bed and illuminated the participant's head with 850 nm light, providing non-intrusive illumination for night-time monitoring. A video was recorded before, during and following an apnea event at 60 fps, yielding 17 ms temporal resolution. Per-pixel absorbance signals were extracted using a Beer-Lambert derived light transport model, and subsequently denoised. The extracted hemodynamic signal exhibited dynamic temporal modulation during and following the apnea event. In particular, the pulse wave amplitude (PWA) decreased during obstructed breathing, indicating vasoconstriction. Upon successful inhalation, the PWA gradually increased toward homeostasis following a temporal phase delay. This temporal vascular tone modulation provides insight into autonomic and vascular response, and may be used to assess sleep apnea using non-contact biophotonic imaging.

Currently, the cardiac activity of infants in the Neonatal Intensive Care Unit (NICU) is monitored with contact sensors. These techniques can cause injuries and infections, particularly in very premature infants with fragile skin. Recently, remote photoplethysmography (rPPG) showed its potential to measure cardiac activity with a camera without skin contact. The main limitations of this technique are its lack of robustness to subject motion and visible light requirements. The aim of this study is to investigate the feasibility of robust rPPG for NICU patients in near darkness. Video recordings using dedicated infrared illumination were made of 7 infants, age 30-33 weeks, at a NICU in Eindhoven, The Netherlands. The pulse rate can be detected with an average error of 1.5 BPM and 2.1 BPM when measured at the face and upper torso region, respectively. Overall, the correct pulse rate is detected for 87% of the time. A camera-based framework for robust pulse extraction in near darkness of NICU patients was proposed and successfully validated. The pulse rate could be reliably detected from all evaluated skin regions. Recordings with vigorous body movements, involving occlusion of the selected skin region, are still a challenge.

To effectively capture human vital signs, a multi-wavelength optoelectronic patch sensor (MOEPS), together with a schematic architecture of electronics, was developed to overcome the drawbacks of present photoplethysmographic (PPG) sensors. To obtain a better performance of in vivo physiological measurement, the optimal illuminations, i.e., light emitting diodes (LEDs) in the MOEPS, whose wavelength is automatically adjusted to each specific subject, were selected to capture better PPG signals. A multiplexed electronic architecture has been well established to properly drive the MOEPS and effectively capture pulsatile waveforms at rest. The protocol was designed to investigate its performance with the participation of 11 healthy subjects aged between 18 and 30. The signals obtained from green (525nm) and orange (595nm) illuminations were used to extract heart rate (HR) and oxygen saturation (SpO₂%). These results were compared with data, simultaneously acquired, from a commercial ECG and a pulse oximeter. Considering the difficulty for current devices to attain the SpO2%, a new computing method, to obtain the value of SpO2%, is proposed depended on the green and orange wavelength illuminations. The values of SpO₂% between the MOEPS and the commercial Pulse Oximeter devics showed that the results were in good agreement. The values of HR showed close correlation between commercial devices and the MOEPS (HR: r1=0.994(Green); r2=0.992(Orange); r3=0.975(Red); r₄=0.990(IR)).

We propose a method to estimate transcutaneous bilirubin, hemoglobin, and melanin based on the diffuse reflectance spectroscopy. In the proposed method, the Monte Carlo simulation-based multiple regression analysis for an absorbance spectrum in the visible wavelength region (460-590 nm) is used to specify the concentrations of bilirubin (C_bil), oxygenated hemoglobin (C_oh), deoxygenated hemoglobin (C_dh), and melanin (C_m). Using the absorbance spectrum calculated from the measured diffuse reflectance spectrum as a response variable and the extinction coefficients of bilirubin, oxygenated hemoglobin, deoxygenated hemoglobin, and melanin, as predictor variables, multiple regression analysis provides regression coefficients. Concentrations of bilirubin, oxygenated hemoglobin, deoxygenated hemoglobin, and melanin, are then determined from the regression coefficients using conversion vectors that are numerically deduced in advance by the Monte Carlo simulations for light transport in skin. Total hemoglobin concentration (C_th) and tissue oxygen saturation (StO₂) are simply calculated from the oxygenated hemoglobin and deoxygenated hemoglobin. In vivo animal experiments with bile duct ligation in rats demonstrated that the estimated C_bil is increased after ligation of bile duct and reaches to around 20 mg/dl at 72 h after the onset of the ligation, which corresponds to the reference value of C_bil measured by a commercially available transcutaneous bilirubin meter. We also performed in vivo experiments with rats while varying the fraction of inspired oxygen (FiO₂). C_oh and C_dh decreased and increased, respectively, as FiO₂ decreased. Consequently, StO₂ was dramatically decreased. The results in this study indicate potential of the method for simultaneous evaluation of multiple chromophores in skin tissue.

Capillary refill time (CRT) is a part of the cardiorespiratory examination in dogs. Changes in CRT can reflect pathological conditions like shock or anemia. Visual CRT estimation has low repeatability; therefore, optical systems for automated estimation have recently appeared. Since existing systems are unsuitable for use in dogs, we designed a simple, small and portable device, which could be easily used at veterinary clinic. The device was preliminarily tested on several measurement sites in two dogs. Not all measurement sites were suitable for CRT measurements due to underlying tissue optical and mechanical properties. The CRT measurements were possible on the labial mucosa, above the sternum and on the digit where CRT was in the range of values, retrieved from the color video of the visual CRT measurement. It seems that light penetration predominantly governs tissue optical response when the pressure is applied. Therefore, it is important to select a proper light, which reaches only superficial capillaries and does not penetrate deeper. Blue or green light is probably suitable for light skin or mucosa, on the other hand, red or near-infrared might be used for skin with pigmented or thick epidermis. Additionally, further improvements of the device design are considered, like adding a calibrated spring, which would insure application of consistent pressure.

A multimodality endoscope system has been designed for early detection of ovarian cancer. Multiple illumination and detection systems must be integrated in a compact, stable, transportable configuration to meet the requirements of a clinical setting. The proximal configuration presented here supports visible light navigation with a large field of view and low resolution, high resolution multiphoton microscopy (MPM), and high resolution optical coherence microscopy (OCM). All modalities are integrated into a single optical system in the endoscope. The system requires two light sources: a green laser for visible light navigation and a compact fiber based femtosecond laser for MPM and OCM. Using an inline wavelength division multiplexer, the two sources are combined into a single mode fiber. To accomplish OCM, a fiber coupler is used to separate the femtosecond laser into a reference arm and signal arm. The reflected reference arm and the signal from the sample are interfered and wavelength separated by a reflection grating and detected using a linear array. The MPM signal is collimated and goes through a series of filters to separate the 2nd and 3rd harmonics as well as twophoton excitation florescence (2PEF) and 3PEF. Each signal is independently detected on a photo multiplier tube and amplified. The visible light is collected by multiple high numerical aperture fibers at the endoscope tip which are bundled into one SMA adapter at the proximal end and connected to a photodetector. This integrated system design is compact, efficient and meets both optical and mechanical requirements for clinical applications.

We developed an optical detector of thrombus formed on the pivot bearing of an extracorporeal centrifugal blood pump (MERA HCF-MP23; Senko Medical Instrument Mfg. Co., Ltd., Tokyo, Japan) which is frequently used for long-term extracorporeal circulation support to bridge to an implantable artificial heart, which in turn is used for bridge to heart transplantation in Japan. In this study, we investigated the quantitative performance of the thrombus formation in acute animal experiments. A total of three experiments of extracorporeal left ventricular assist using Japanese specific pathogen-free pigs were conducted. The optical fibers were set in the pump driver unit. The incident light at nearinfrared wavelength aiming at the pivot bearing and the resulting scattered light were guided to respective fibers. The detected signal was analyzed to obtain thrombus formation level (TFL) calculated by a specially developed software. When the increase in TFL was confirmed, the pump was exchanged and the extracorporeal circulation was restarted. The number of pump exchanges were four times at each experiment so a total of twelve pumps were evaluated. 3-dimentional data surrounding the pivot bearing and the adhered thrombus was captured by a 3-dimantional surface measurement system to calculate the thrombus surface area (TSA) formed on the pivot bearing. As a result, the correlation coefficient between TFL and TSA was 0.878. The accuracy of TSA estimated by the optical detector was 3.6±2.3 mm². This was small enough to not have the pump exchanged in clinical judgement. The developed detector would be useful for optimal anti-coagulation management.

Major advancements in the fields of electronics, photonics and wireless communication have enabled the development of compact wearable devices, with applications in diverse domains such as fitness, wellness and medicine. In parallel, nanotechnology is enabling the development of miniature sensors that can detect events at the nanoscale with unprecedented accuracy. On this matter, in vivo implantable Surface Plasmon Resonance (SPR) nanosensors have been proposed to analyze circulating biomarkers in body fluids for the early diagnosis of a myriad of diseases, ranging from cardiovascular disorders to different types of cancer. In light of these results, in this paper, an architecture is proposed to bridge the gap between these two apparently disjoint paradigms, namely, the commercial wearable devices and the advanced nano-biosensing technologies. More specifically, this paper thoroughly assesses the feasibility of the wireless optical intercommunications of an SPR-based nanoplasmonic biochip -implanted subcutaneously in the wrist-, with a nanophotonic wearable smart band which is integrated by an array of nano-lasers and photon-detectors for distributed excitation and measurement of the nanoplasmonic biochip. This is done through a link budget analysis which captures the peculiarities of the intra-body optical channel at (sub) cellular level, the strength of the SPR nanosensor reflection, as well as the capabilities of the nanolasers (emission power, spectrum) and the nano photon-detectors (sensitivity and noise equivalent power). The proposed analysis guides the development of practical communication designs between the wearable devices and nano-biosensing implants, which paves the way through early-stage diagnosis of severe diseases.

Here a photonic waveguide on Al₂O₃/SiO₂ platform is proposed to cover the 240~320 nm wavelength-range, which is of paramount significance in protein and nuclei acid quantification. Our optical waveguide increases path-length and overlap integration for light-matter interaction with proteins. The proposed system detects one order less proteins concentration as low as 12.5 μg/ml compared with NanoDrop^TM that detects <125 μg/ml. Also, a linear absorbance change up to protein concentration of 7500 μg/ml is experimentally attained which is based on the Beer-Lambert-law.

The main research objective is the development of innovative optical technologies for sensitive diagnosis of early stages of development of stomach cancer and monitoring of stress-induced appearance and development of tumors of the gastrointestinal tract by applying endogenous and exogenous fluorescence spectroscopy modalities.

Different mechanisms solely and in combination for evaluation of the joint impact of bioenvironmental factors (stress, Helicobacter pillory, exo-toxins in the food, water, soil and air) were applied to induce gastrointestinal tract (GIT) neoplasia in rats. The transformation of damaged areas of the stomach mucosa into malignancies in all parts of gastrointestinal tract were detected using exogenous fluorescence of photosensitizers - 5-aminolevulinic acid (5-ALA) and aluminum phthalocyanine (Al-Pc). Fluorescent mapping of different organs (liver, spleen, lungs, brain) also was developed – to evaluate the distribution of the photosensitizers in the whole body on the second hour after photosensitizer application by intravenous injection. Fiber-optic probe was used to measure the organs investigated. Fluorescence spectra were detected by microspectrometer USB4000 (OceanOptics Inc., USA), and FS405 LED source on 405 nm was used as excitation source for both types of photosensitizers applied.

Diagnostically-important parameters of oximetry, optical coherence tomography and speckle-imaging of the microcirculation of the stomach were also evaluated, to evaluate changes in the blood flow and vascular architecture, during the formation of the initial phases of the neoplasm development.

The human immunodeficiency virus-1 (HIV-1) is currently detected using conventional qualitative and quantitative tests to determine the presence or absence of HIV in blood samples. However, the approach of these tests detects the presence of either viral antibodies or viral RNA that require labelling which may be costly, sophisticated and time consuming. A label-free approach of detecting the presence of HIV is therefore desirable. Of note optical tweezers can be coupled with other technologies including spectroscopy, which also investigates light-matter interactions. For example, coupling of optical tweezers with luminescence spectroscopy techniques has emerged as a powerful tool in biology for micro-manipulation, detection and analysis of individual cells. Integration of optical techniques has enabled studying biological particles in a label-free manner, whilst detecting functional groups and other essential molecules within mixed populations of cells. In the current study, an optical trapping system coupled to luminescence spectroscopy was utilised to detect the presence of HIV infection in TZM-bl cells in vitro. This was performed by infecting TZM-bl cells with the ZM53 HIV-1 pseudovirus, and incubating them for 48 hours prior analysis. The differences between infected and uninfected cells were thereafter displayed as shown by the spectrographs obtained. Combination of these two techniques has a potential in the field of infectious disease diagnostics.

This study evaluated the polymerization shrinkage of two experimental flowable composite resins (CR) with different proportions of Urethane dimethacrylate (UDMA)/triethylene glycol dimethacrylate (TEGDMA) monomers in the organic matrix (50:50 and 60:40, respectively). A commercially available flowable CR, Tetric N-Flow (Ivoclair Vivadent, Liechtenstein, Germany), was employed as the control group. The resins were inserted in a cylindrical teflon mold (7 mm diameter, 0.6 mm height) and scanned with OCT before photoactivation, immediately after and 15 minutes after light-curing (Radii-Cal, SDI, Australia, 1,200 mW/cm² ) exposure. A Callisto SD-OCT system (Thorlabs Inc, USA), operating at 930 nm central wavelength was employed for imaging acquisition. Cross-sectional OCT images were captured with 8 mm transverse scanning (2000x512 matrix), and processed by the ImageJ software, for comparison between the scanning times and between groups. Pearson correlation showed significant shrinkage for all groups in each time analyzed. Kruskal-Wallis test showed greater polymerization shrinkage for the 50:50 UDMA/TEGDMA group (p=0.001), followed by the control group (p=0.018). TEGDMA concentration was proportionally related to the polymerization shrinkage of the flowable composite resins.

The BioCD platform technology uses spinning-disk interferometry to detect molecular binding to target molecular probes in biological samples. Interferometric configurations have included differential phase contrast and in-line quadrature detection. For the detection of extremely low analyte concentrations, nano- or microparticles can enhance the signal through background-free diffraction detection. Diffraction signal measurements on BioCD biosensors are achieved by forming gratings on a disc surface. The grating pattern was printed with biotinylated bovine serum albumin (BSA) and streptavidin coated beads were deployed. The diameter of the beads was 1 micron and strong protein bonding occurs between BSA and streptavidin-coated beads at the printed location. The wavelength for the protein binding detection was 635 nm. The periodic pattern on the disc amplified scattered light into the first-order diffraction position. The diffracted signal contains Mie scattering and a randomly-distributed-bead noise contributions. Variation of the grating pattern periodicity modulates the diffraction efficiency. To test multiple spatial frequencies within a single scan, we designed a fan-shaped grating to perform frequency filter multiplexing on a diffraction-based BioCD.

Recent developments in understanding of nanomaterial behaviors and synthesis have led to their application across a wide range of commercial and scientific applications. Recent investigations span from applications in nanomedicine and the development of novel drug delivery systems to nanoelectronics and biosensors. In this study, we propose the application of a newly engineered temperature sensitive water-based bio-compatible core/shell up-conversion nanoparticle (UCNP) in the development of a smart substrate for remote temperature sensing. We developed this smart substrate by dispersing functionalized nanoparticles into a polymer solution and then spin-coating the solution onto one side of a microscope slide to form a thin film substrate layer of evenly dispersed nanoparticles. By using spin-coating to deposit the particle solution we both create a uniform surface for the substrate while simultaneously avoid undesired particle agglomeration. Through this investigation, we have determined the sensitivity and capabilities of this smart substrate and conclude that further development can lead to a greater range of applications for this type smart substrate and use in remote temperature sensing in conjunction with other microscopy and spectroscopy investigations.

In this paper, a comparative study between SVM and HMM has been carried out for multiclass classification of cervical healthy and cancerous tissues. In our study, the HMM methodology is more promising to produce higher accuracy in classification.