This PDF file contains the front matter associated with SPIE Proceedings Volume 9332, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and Conference Committee listing.

Attenuated total reflection (ATR)-FTIR spectroscopy is a convenient technique for analysing biomedical samples because of its sensitivity to subtle compositional changes, speed of data acquisition and ease of sample preparation. We have applied the technology to the detection of disease biomarkers in urine and investigated the translation of these diagnostic methods to simple bench-top spectrometers. To demonstrate the use of ATR-FTIR spectroscopy as a bedside diagnostic tool, we have installed a roomtemperature bench-top infrared spectrometer in the renal unit at the Royal Free Hospital (RFH), London. A nurse recorded spectra of urine from patients with a range of conditions, including diabetes, kidney disease, stone disease and urinary tract infections, and the data were correlated to medical conditions to assess the diagnostic capabilities of the system and to identify potential spectral patterns associated with disease. Two hundred and six spectra have been recorded to date; these show it is possible to detect urea, creatinine, protein, lipids, sugars and other minor metabolites, including potential disease biomarkers. Several spectral peaks of potential diagnostic interest were identified that show variations between normal and disease samples.

Surface Enhanced Raman spectroscopy (SERS) offers sensitive and non-invasive detection of a variety of compounds as well as unparalleled information for establishing the molecular identity of both inorganic and organic compounds, not only in biological fluids but in all other aqueous and non-aqueous media. The localized hotspots produced through SERS at the solution/nanostructure interface of clustered gold or silver nano-particles enables detection levels of parts per trillion. Recent developments in advanced fabrication methods have enabled the manufacture of SERS substrates with repeatable surface nanostructures which provide reproducible quantitative analysis, historically a weakness of the SERS technique. In this paper we describe the novel use of gold sputtered Blu-Ray surfaces as SERS substrates. Blu-Ray disks provide ideal surfaces of SERS substrates with their repeatable and regular nano-gratings. We show that the unique surface features and composition of the recording surface enables the formation of gold nano-islands with nanogaps, simply through gold sputtering, and relate this to a 600 fold signal increase of the melamine Raman signal in aqueous solutions and detection to 68 ppb. Melamine is a triazine compound and appears not only as environmental contaminant in environmental groundwater but also as an adulterant in foods due to its high nitrogen content. We have shown significant SERS signal enhancements for spectra of melamine using gold-sputtered Blu-Ray disk surfaces, with reproducibility of 12%. Blu-Ray disks have a unique combination of design, surface features and composition of the recording surface which makes them ideal for preparation of SERS substrates by gold sputter-coating.

Competitive binding assays comprised of the protein Concanavalin A (ConA) have shown potential for use in continuous glucose monitoring devices. However, its time-dependent, thermal instability can impact the lifetime of these ConA based assays. In an attempt to design sensors with longer in vivo lifetimes, different groups have immobilized the protein to various surfaces. For example, Ballerstadt et al. have shown that immobilizing ConA onto the interior of a micro-dialysis membrane and allowing dextran to be freely suspended within solution allowed for successful in vivo glucose sensing up to 16 days. This work explores the glucose response of an assay comprised of modified ConA and a single fluorescently labeled competing ligand in free solution to increase the in vivo sensing lifetime without immobilization,. The behavior of this assay in the presence of varying glucose concentrations is monitored via fluorescence anisotropy over a 30 day period.

Micro-dialysis has been established in the clinical environment for continuously harvesting body fluids, but a drawback of this process are variable recovery rates, which can be observed especially for subcutaneously implanted catheters. Perfusates with either acetate or mannitol have been investigated as recovery markers. The latter substance is suggested for application with external cavity tuneable quantum cascade lasers, rendering a limited wavenumber interval in contrast to FTIR-spectrometers. Despite the overlap of mannitol and glucose spectra, their simultaneous quantification was successful. By investigating the depletion of the marker substances from the perfusates using different micro-dialysis devices, the theoretical nonlinear relationship between the relative dialysate marker concentration and glucose recovery rate was confirmed for the marker substance-analyte pair of acetate and glucose, rendering a basis for reliable blood glucose measurements. For the pair of mannitol and glucose an almost linear dependency was expected for the microdialysate catheters and experimentally verified, which provides a straightforward correction of any dialysis recovery rate variation during patient monitoring.

We have developed an instrument for real time continuous interferometric measurement of dimensional change in stimuli-responsive hydrogel sensors. The instrument consists of a superfluorecent LED and a low-cost grating spectrometer. All signal conditioning and processing is performed using a microprocessor. We have tested the instrument with a glucose sensitive hydrogel sensor. The rms noise on the length measurement is less than 1.4 nm, whereas rms noise on the length change is less than 0.25 nm. An observed bias of the length measurement can be reduced by minor changes in the frequency estimation algorithm used.

A point of care biosensor capable of detecting biomarkers in a low concentration in blood samples can have a great impact on the healthcare community. Surface Enhanced Raman Spectroscopy (SERS) is one potential means of monitoring analytes at low concentrations. Toward a continuing effort to use SERS for point of care biosensing, in this paper spherical gold colloid and nanocages are analyzed and compared to determine which substrate has better utility. Gold colloid is and has been a popular substrate used in SERS. However, for biosensing its use can be problematic since aggregates must be formed typically using salt, which are time dependent, fall out of solution, and generally do not provide good reproducibility. Thus, in this work, nanocages are analyzed and compared as an alternative to using gold colloid for quantifiable SERS biosensing. Scanning electron microscope (SEM) and transverse electron microscope (TEM) images are depicted for each material along with their extinction coefficients, aggregation properties, and SERS spectrum both with and without salt added. Overall, nanocages are shown to provide equivalent SERS enhancement without the need for salt induced aggregation and hence have the potential to be a better substrate for reproducible SERS biosensing.

Surface-enhanced Raman spectroscopy (SERS) is a spectroscopic technique, where Raman scattering is boosted primarily by enhanced electric field due to localized surface plasmon resonance (LSPR). With advances in nanofabrication techniques, SERS has attracted great attention for label-free molecular sensing and imaging. However, the practical use of SERS has often encountered an inherent issues regarding a molecule transfer step where target molecules need to be within the close proximity of a SERS-active surface by either mixing with nanoparticles or coating onto surface-bound nanostructures. To address this issue, we have developed stamping surface-enhanced Raman spectroscopy (S-SERS) for label-free, multiplexed, molecular sensing and large-area, high-resolution molecular imaging on a flexible, non-plasmonic surface without solution-phase molecule transfer. In this technique, a polydimethylsiloxane (PDMS) thin film and nanoporous gold disk SERS substrate play the roles as molecule carrier and Raman signal enhancer, respectively. After stamping the SERS substrate onto the PDMS film, SERS measurements can be directly taken from the “sandwiched” target molecules. The performance of S-SERS is evaluated by the detection of Rhodamine 6G (R6G), urea, and its mixture with acetaminophen (APAP), in physiologically relevant concentration range, along with corresponding SERS spectroscopic maps. S-SERS features simple sample preparation, low cost, and high reproducibility, which could lead to SERS-based sensing and imaging for point-of-care and forensics applications.

Coreshell plasmonic nanoparticles (CS) are a class of nanoparticles that exhibit optical absorption in the near IR regime and have potential biomedical applications in imaging, therapy and sensing. We present our preliminary investigation on the applications of CS as a surface plasmon based sensor to study the functional properties of human blood. CS particles of size about 1 μm exhibit broad absorption between 650 nm to 1000 nm, the regime generally used to study blood saturation. We synthesized CS particles of size about 1μm, coated with a thin shell. The core medium was polystyrene and the nano-shell layer was gold. The plasmon peak of CS varied with blood concentration. The study showed that 750 nm plasmonic peak of CS exhibits the wavelength shift of 4.11±0.26 nm per hematocrit.

A novel multi-wavelength photoplethysmograph (PPG), previously utilized to quantify optically absorptive circulating gold nanoparticles, has demonstrated the potential to enhance therapeutic treatment predictability as pharmacokinetic metrics are provided throughout the intravenous delivery and clearance phase of amphotericin b (injected in the lipid form Abelcet®) in real-time. This report demonstrates how the PPG could be used to assess the real-time bioavailability of intravenously delivered optically-absorbing therapeutic agents. The drug currently under investigation is antifungal amphotericin b (absorption peak ~355 nm). We describe how the algorithm has been adapted to quantify the concentration of amphotericin b in the pulsatile, circulating blood based on its extinction at three wavelengths (355, 660 and 940 nm) corresponding to the peaks of amphotericin b and wavelengths for oxygen saturation measurements, respectively. We show an example of the system collecting data representing the baseline, injection, and the clearance phases. The PPG device showed a measurement range of concentrations between 0.0987 mg/mL to 0.025mg/ml in blood. An examination of the data obtained suggests that the system is well suited to sense the concentration of amphotericin b at a therapeutic dose (≈5 mg/kg/day).

The aim of the current work is to investigate the possibility of augmenting pulse oximetry algorithms to enable the estimation of venous parameters in peripheral tissues. In order to further understand the contribution of venous blood to the photoplethysmographic (PPG) signal, recordings were made from six healthy volunteer subjects during an exercise in which the right hand was placed in various positions above and below heart level. The left hand was kept at heart level as a control while the right hand was moved. A custom-made two-channel dual wavelength PPG instrumentation system was used to obtain the red and infrared plethysmographic signals from both the right and left index fingers simultaneously using identical sensors. Laser Doppler flowmetry signals were also recorded from an adjacent fingertip on the right hand. Analysis of all acquired PPG signals indicated changes in both ac and dc amplitude of the right hand when the position was changed, while those obtained from the left (control) hand remained relatively constant. Most clearly, in the change from heart level to 50cm below heart level there is a substantial decrease in both dc and ac amplitudes. This decrease in dc amplitude most likely corresponds to increased venous pooling, and hence increased absorption of light. It is speculated that the decrease in ac PPG amplitude is due to reduced arterial emptying during diastole due to increased downstream resistance due to venous pooling.

The paper will describe the novel multi-wavelength photometric device OxyTrue Hb® which is capable to measure the hemoglobin (Hb) and methemoglobin (MetHb) concentration non-invasively. Clinic trails in blood donation centers and during the dialysis are done to prove and demonstrate the performance of the system. The results are compared to the gold standard, the BGA measurement.

Acute scrotum is a urologic condition defined by scrotal pain, swelling, and redness of acute onset. Prompt diagnosis and treatment are necessary to preserve testicular viability. The history and clinical symptoms reported are key to diagnosis and proper treatment, but are not always readily obtained in children, in whom common causes of acute scrotum include testicular torsion, torsion of the appendix testis, and epididymitis. These acute conditions have different causal pathology that mandate specific treatment, hence the importance of early and accurate diagnosis.

Integration of optical imagers and sensors into recently emerging wearable computational devices allows for simpler and more intuitive methods of integrating biomedical imaging and medical diagnostics tasks into existing infrastructures. Here we demonstrate the ability of one such device, the Google Glass, to perform qualitative and quantitative analysis of immunochromatographic rapid diagnostic tests (RDTs) using a voice-commandable hands-free software-only interface, as an alternative to larger and more bulky desktop or handheld units. Using the built-in camera of Glass to image one or more RDTs (labeled with Quick Response (QR) codes), our Glass software application uploads the captured image and related information (e.g., user name, GPS, etc.) to our servers for remote analysis and storage. After digital analysis of the RDT images, the results are transmitted back to the originating Glass device, and made available through a website in geospatial and tabular representations. We tested this system on qualitative human immunodeficiency virus (HIV) and quantitative prostate-specific antigen (PSA) RDTs. For qualitative HIV tests, we demonstrate successful detection and labeling (i.e., yes/no decisions) for up to 6-fold dilution of HIV samples. For quantitative measurements, we activated and imaged PSA concentrations ranging from 0 to 200 ng/mL and generated calibration curves relating the RDT line intensity values to PSA concentration. By providing automated digitization of both qualitative and quantitative test results, this wearable colorimetric diagnostic test reader platform on Google Glass can reduce operator errors caused by poor training, provide real-time spatiotemporal mapping of test results, and assist with remote monitoring of various biomedical conditions.

The growing power of imaging and computing power of smartphones is creating the possibility of converting your smartphone into a high power pocket microscopy system. High quality miniature microscopy lenses attached to smartphone are typically made with glass or plastics that can only be produce at low cost with high volume. To revise the paradigm of microscope lenses, we devised a simple droplet lens fabrication technique that which produces low cost and high performance lens. Each lens is integrated into thin 3-D printed holder with complimentary light emitted diode (LEDs) that clips onto majority of smartphones. The integrated device converts a smartphone into a high power optical microscope/dermatoscope at around $2. This low cost device has wide application in a multitude of practical uses such as material inspection, dermascope and educational microscope.

Many cases of epithelial cancer originate in basal layers of tissue and are initially undetected by conventional microendoscopy techniques. We present a bench-top, fiber-bundle microendoscope capable of providing high resolution images of surface cell morphology. Additionally, the microendoscope has the capability to interrogate deeper into material by using diffuse reflectance and broadband diffuse reflectance spectroscopy. The purpose of this multimodal technique was to overcome the limitation of microendoscopy techniques that are limited to only visualizing morphology at the tissue or cellular level. Using a custom fiber optic probe, high resolution surface images were acquired using topical proflavine to fluorescently stain non-keratinized epithelia. A 635 nm laser coupled to a 200 μm multimode fiber delivers light to the sample and the diffuse reflectance signal was captured by a 1 mm image guide fiber. Finally, a tungsten-halogen lamp coupled to a 200 μm multimode fiber delivers broadband light to the sample to acquire spectra at source-detector separations of 374, 729, and 1051 μm. To test the instrumentation, a high resolution proflavine-induced fluorescent image of resected healthy mouse colon was acquired. Additionally, five monolayer poly(dimethylsiloxane)-based optical phantoms with varying absorption and scattering properties were created to acquire diffuse reflectance profiles and broadband spectra.

An FTIR image of an 8 µm section of de-paraffinised bronchial biopsy that shows a histological transition from normal to severe dysplasia/squamous cell carcinoma (SCC) in situ was obtained in transmission by stitching together images of 256 x 256 µm recorded using a 96 x 96 element FPA detector. Each pixel spectrum was calculated from 128 co-added interferograms at 4 cm⁻¹ resolution. In order to improve the signal to noise ratio, blocks of 4x4 adjacent pixels were subsequently averaged. Analyses of this spectral image, after conversion of the spectra to their second derivatives, show that the epithelium and the lamina propria tissue types can be distinguished using the area of troughs at either 1591, 1334, 1275 or 1215 cm⁻¹ or, more effectively, by separation into two groups by hierarchical clustering (HCA) of the 1614-1465 region. Due to an insufficient signal to noise ratio, disease stages within the image could not be distinguished with this extent of pixel averaging. However, after separation of the cell types, disease stages within either the epithelium or the lamina propria could be distinguished if spectra were averaged from larger, manually selected areas of the tissue. Both cell types reveal spectral differences that follow a transition from normal to cancerous histology. For example, spectral changes that occurred in the epithelium over the transition from normal to carcinoma in situ could be seen in the 1200-1000 cm⁻¹ region, particularly as a decrease in the second derivative troughs at 1074 and 1036 cm⁻¹ , consistent with changes in some form of carbohydrate. Spectral differences that indicate a disease transition from normal to carcinoma in the lamina propria could be seen in the 1350-1175 cm⁻¹ and 1125-1030 cm⁻¹ regions. Thus demonstrating that a progression from healthy to severe dysplasia/squamous cell carcinoma (SCC) in situ can be seen using FTIR spectroscopic imaging and multivariate analysis.

We present a visualization pipeline from sample to answer for point-of-care blood cell counting applications. Effective and low-cost point-of-care medical diagnostic tests provide developing countries and rural communities with accessible healthcare solutions [1], and can be particularly beneficial for blood cell count tests, which are often the starting point in the process of diagnosing a patient [2]. The initial focus of this work is on total white and red blood cell counts, using a microfluidic cartridge [3] for sample processing. Analysis of the processed samples has been implemented by means of two main optical visualization systems developed in-house: 1) a fluidic operation analysis system using high speed video data to determine volumes, mixing efficiency and flow rates, and 2) a microscopy analysis system to investigate homogeneity and concentration of blood cells. Fluidic parameters were derived from the optical flow [4] as well as color-based segmentation of the different fluids using a hue-saturation-value (HSV) color space. Cell count estimates were obtained using automated microscopy analysis and were compared to a widely accepted manual method for cell counting using a hemocytometer [5]. The results using the first iteration microfluidic device [3] showed that the most simple – and thus low-cost – approach for microfluidic component implementation was not adequate as compared to techniques based on manual cell counting principles. An improved microfluidic design has been developed to incorporate enhanced mixing and metering components, which together with this work provides the foundation on which to successfully implement automated, rapid and low-cost blood cell counting tests.

Hypercholesterolemia is characterized by high levels of cholesterol in the blood and is associated with an increased risk of atherosclerosis and coronary heart disease. Early detection of hypercholesterolemia is necessary to prevent onset and progress of cardiovascular disease. Optical imaging techniques might have a potential for early diagnosis and monitoring of hypercholesterolemia. In this study, hyperspectral imaging was investigated for this application. The main aim of the study was to identify spectral and spatial characteristics that can aid identification of hypercholesterolemia in facial skin. The first part of the study involved a numerical simulation of human skin affected by hypercholesterolemia. A literature survey was performed to identify characteristic morphological and physiological parameters. Realistic models were prepared and Monte Carlo simulations were performed to obtain hyperspectral images. Based on the simulations optimal wavelength regions for differentiation between normal and cholesterol rich skin were identified. Minimum Noise Fraction transformation (MNF) was used for analysis. In the second part of the study, the simulations were verified by a clinical study involving volunteers with elevated and normal levels of cholesterol. The faces of the volunteers were scanned by a hyperspectral camera covering the spectral range between 400 nm and 720 nm, and characteristic spectral features of the affected skin were identified. Processing of the images was done after conversion to reflectance and masking of the images. The identified features were compared to the known cholesterol levels of the subjects. The results of this study demonstrate that hyperspectral imaging of facial skin can be a promising, rapid modality for detection of hypercholesterolemia.

Near-infrared spectroscopy has a potential for noninvasive determination of skin moisture level due to high water absorption. In this study, diffuse reflectance spectra of in vivo skin were acquired in the spectral range of 900 nm to 1700 nm by using near-infrared spectrometer, optical fiber and halogen bulb light source. Absorption changes after applying skin moisturizers were analyzed over time at different body sites. Results show difference in absorption when comparing dry and normal skin. Comparison of absorption changes over time after applying moisturizer at different body sites is analyzed and discussed. Some patterns of how skin reacts to different skin moisturizers are shown, although no clear pattern can be seen due to signal noise.

We proposed a low cost optical cavity based biosensor with a differential detection for point-of-care diagnosis. Two lasers at different wavelengths are used for the differential detection. This method enhances the sensitivity through higher responsivity and noise cancelation. To reduce noise further, especially due to the unstable low cost laser diode output, we employed a referencing method in which a reference pixel value in each CMOS image frame is subtracted from all other pixels. To validate the designed structure and demonstrate the sensitivity of it, we perform refractive index measurements of fluids with our design. In this presentation, we will discuss our design, simulation results, and measurement results.

Therapeutic drug monitoring (TDM) contributes to safe and effective pharmacotherapy in clinical fields. A simple, rapid, low-cost, and minimally-invasive drug measurement method attracts much interest for point-of-care TDM. Tear fluids can be collected minimally-invasively compared to blood sampling and there is a correlation between a drug concentration in tears and that in bloods. Surface enhanced Raman spectroscopy (SERS) with paper-based substrate is useful for point-of-care TDM owing to inexpensiveness and high-sensitivity. Paper is also a safe tear collection tool. Then we are studying on a paper-based SERS of tear specimen for point-of-care TDM. In this paper, to improve sensitivity in measuring drug concentration in tear fluids, we fabricated a SERS substrate by coating gold nano-rods on a paper substrate and evaluated whether the fabricated substrate can enhance Raman scattering. Sodium phenobarbital (PB), an anti-convulsant agent, was used as a target. In experiment, the fabricated substrate indicated the lower detection limit of PB in a solution than a plain paper substrate. This result showed the potential of the paper based SERS substrate to measure drug concentration in tears simply and inexpensively.

LIBS has been proven to be a robust elemental analysis tool attracting interest because of the wide applications. LIBS can be used for analysis of any type of samples i.e. environmental/physiological, regardless of its state of matter. Conventional spectroscopy techniques are good in analytical performance, but their sample preparation method is mostly destructive and time consuming. Also, almost all these methods are incapable of analysing multi elements simaltaneously. On the other hand, LIBS has many potential advantages such as simplicity in the experimental setup, less sample preparation, less destructive analysis of sample etc. In this paper, we report some of the biomedical applications of LIBS. From the experiments carried out on clinical samples (calcified tissues or teeth and gall stones) for trace elemental mapping and detection, it was found that LIBS is a robust tool for such applications. It is seen that the presence and relative concentrations of major elements (calcium, phosphorus and magnesium) in human calcified tissue (tooth) can be easily determined using LIBS technique. The importance of this study comes in anthropology where tooth and bone are main samples from which reliable data can be easily retrieved. Similarly, elemental composition of bile juice and gall stone collected from the same subject using LIBS was found to be similar. The results show interesting prospects for LIBS to study cholelithiasis (the presence of stones in the gall bladder, is a common disease of the gastrointestinal tract) better.

Multivariate Optical Elements (MOEs) are thin-film devices that encode a broad band, spectroscopic pattern allowing a simple broadband detector to generate a highly sensitive and specific detection for a target analyte. MOE filter sets are capable of sensing projections of the original sparse spectroscopic space enabling a small set of MOEs to discriminate a multitude of target analytes. This presentation will summarize the design and fabrication of compressed detection MOE filter sets for detecting multiple fluorochromes simultaneously with strong spectroscopic interference as well as comparing the detection performance of the MOE sensor with traditional optical band pass filter methodologies.

A quantitative comparison has been performed between two commercial near-infrared (NIR) vein-viewing systems which are designed to supplement the clinician’s traditional skills in locating veins by means of visualization and palpation. The AccuVein AV300 and Novarix IV-eye real-time imaging systems employ very different imaging geometries; the former generates an image from reflected NIR light produced by a beam scanned across the surface, while the latter illuminates the viewed region at four points on the periphery and records the resulting distribution of diffusely transmitted light. The comparison involved measuring the contrast produced by absorbing rods (simulated blood vessels) in a cylindrical phantom with tissue-like optical properties, and the contrast of superficial blood vessels in the arms of healthy volunteers. The locations and sizes of the blood vessels were independently verified using a clinical ultrasound imaging system. The phantom measurements suggested that the AV300 displays the most superficial vessels with greater contrast, but the IV-eye is able to detect vessels when they are at a depth up to 2 mm greater than the limit observed for the AV300. The results for thirty healthy volunteers also indicated that the AV300 typically displays vessels with higher overall contrast, but the effectiveness of the IV-eye at visualizing deeper vessels was even more pronounced, with a maximum depth several millimeters greater than that achieved by the AV300, and more than ten times as many vessels observed at depths below 4 mm.