This PDF file contains the front matter associated with SPIE Proceedings Volume 9701, including the Title Page, Copyright information, Table of Contents, Introduction, and the Conference Committee listing.

Principal Component Analysis (PCA) is a technique of multivariate data analysis widely used in various fields like biology, ecology or economy to reduce data dimensionality while retaining most important information. It is becoming a standard practice in multispectral/hyperspectral imaging since those multivariate data generally suffer from a high redundancy level.

Nevertheless, by definition, PCA is meant to be applied to a single multispectral/hyperspectral image at a time. When several images have to be treated, running a PCA on each image would generate specific reduced spaces, which is not suitable for comparison between results. Thus, we focus on two PCA based algorithms that could define common reduced spaces of representation. The first method arises from literature and is computed with the barycenter covariance matrix. On the contrary, we designed the second algorithm with the idea of correcting standard PCA using permutations and inversions of eigenvectors.

These dimensionality reduction methods are used within the context of a cosmetological study of a foundation make-up. Available data are in-vivo multispectral images of skin acquired on different volunteers in time series. The main purpose of this study is to characterize the make-up degradation especially in terms of texture analysis. Results have to be validate by statistical prediction of time since applying the product.

PCA algorithms produce eigenimages that separately enhance skin components (pores, radiance, vessels...). From these eigenimages, we extract morphological texture descriptors and intent a time prediction. Accuracy of common reduced spaces outperform classical PCA one. In this paper, we detail how PCA is extended to the multiple groups case and explain what are the advantages of common reduced spaces when it comes to study several multispectral images.

When the pleural cavity is opened during the surgery portion of pleural photodynamic therapy (PDT) of malignant mesothelioma, the pleural volume will deform. This impacts the delivered dose when using highly conformal treatment techniques. To track the anatomical changes and contour the lung and chest cavity, an infrared camera–based navigation system (NDI) is used during PDT. In the same patient, a series of computed tomography (CT) scans of the lungs are also acquired before the surgery. The reconstructed three-dimensional contours from both NDI and CTs are imported into COMSOL Multiphysics software, where a finite element-based (FEM) deformable image registration is obtained. The CT contour is registered to the corresponding NDI contour by overlapping the center of masses and aligning their orientations. The NDI contour is considered as the reference contour, and the CT contour is used as the target one, which will be deformed. Deformed Geometry model is applied in COMSOL to obtain a deformed target contour. The distortion of the volume at X, Y and Z is mapped to illustrate the transformation of the target contour. The initial assessment shows that FEM-based image deformable registration can fuse images acquired by different modalities. It provides insights into the deformation of anatomical structures along X, Y and Z-axes. The deformed contour has good matches to the reference contour after the dynamic matching process. The resulting three-dimensional deformation map can be used to obtain the locations of other critical anatomic structures, e.g., heart, during surgery.

There is a strong clinical need for an optical coherence tomography (OCT) system capable of delivering concurrent coagulation light enabling image-guided dynamic laser marking for targeted collection of biopsies, as opposed to a random sampling, to reduce false-negative findings. Here, we present a system based on double-clad fiber (DCF) capable of delivering pulsed laser light through the inner cladding while performing OCT through the core. A previously clinically validated commercial OCT system (NVisionVLE, Ninepoint Medical) was adapted to enable in vivo esophageal image-guided dynamic laser marking. An optimized DCF coupler was implemented into the system to couple both modalities into the DCF. A DCF-based rotary joint was used to couple light to the spinning DCF-based catheter for helical scanning. DCF-based OCT catheters, providing a beam waist diameter of 62μm at a working distance of 9.3mm, for use with a 17-mm diameter balloon sheath, were used for ex vivo imaging of a swine esophagus. Imaging results using the DCF-based clinical system show an image quality comparable with a conventional system with minimal crosstalk-induced artifacts. To further optimize DCF catheter optical design in order to achieve single-pulse marking, a Zemax model of the DCF output and its validation are presented.

Cervical cancer is the leading cause of cancer death for women in developing countries. Colposcopy plays an important role in early screening and detection of cervical intraepithelial neoplasia (CIN). In this paper, we developed a multimodal colposcopy system that combines multispectral reflectance, autofluorescence, and RGB imaging for in vivo detection of CIN, which is capable of dynamically recording multimodal data of the same region of interest (ROI). We studied the optical properties of cervical tissue to determine multi-wavelengths for different imaging modalities. Advanced algorithms based on the second derivative spectrum and the fluorescence intensity were developed to differentiate cervical tissue into two categories: squamous normal (SN) and high grade (HG) dysplasia. In the results, the kinetics of cervical reflectance and autofluorescence characteristics pre and post acetic acid application were observed and analyzed, and the image segmentation revealed good consistency with the gold standard of histopathology. Our pilot study demonstrated the clinical potential of this multimodal colposcopic system for in vivo detection of cervical cancer.

Melanoma skin cancer has one of the highest mortality rates of all types of cancer if not detected at an early stage. The survival rate is highly dependent on its penetration depth, which is commonly determined by histopathology. In this work, we aim at combining optical coherence tomography and optoacoustic as a non-invasive all-optical method to measure the penetration depth of melanoma. We present our recent achievements to setup a handheld multimodal device and also results from first in vivo measurements on healthy and cancerous skin tissue, which are compared to measurements obtained by ultrasound and histopathology.

To study the relationship between the corneal lipid layer and the ocular surface temperature (OST), we conducted a clinical trial for 20 subjects. Subjects were clinically screened prior to the trial. Of the 20 subjects, 15 have Meibomian gland dysfunction (MGD), and 5 have aqueous-deficient dry eye (ADDE). A custom, circularly polarized illumination video tearscope measured the lipid layer thickness of the ocular tear film. A long-wave infrared video camera recorded the dynamic thermal properties of the ocular team film. The results of these two methods were analyzed and compared. Using principal component analysis (PCA) of the lipid layer distribution, we find that the 20 subjects could be categorized into five statistically significant groups, independent of their original clinical classification: thin (6 subjects), medium (5 subjects), medium and homogenous (3 subjects), thick (4 subjects), and very thick (2 subjects) lipids, respectively. We also conducted PCA of the OST data, and recategorized the subjects into two thermal groups by k-means clustering: one includes all ADDE subjects and some MGD subjects; the other includes the remaining MGD subjects. By comparing these two methods, we find that dry eye subjects with thin (≤ 40 nm) lipids have significantly lower OST, and a larger OST drop range, potentially due to more evaporation. However, as long as the lipid layer is not thin (> 40 nm), there is no strong correlation between the lipid layer thickness and heterogeneity and the OST patterns.

Cone beam computed tomography (CBCT) is limited in guiding irradiation for soft tissue targets. As a complementary imaging modality, bioluminescence tomography (BLT) provides strong soft tissue contrast. We developed a dual-use BLT system which consists of an optical assembly, a mobile cart and an independent mouse bed. The system is motorized which can easily dock onto an independent mouse bed operating as a standalone system for longitudinal bioluminescence imaging (BLI)/BLT studies and also dock onto the SARRP for on-line radiation guidance.

Our initial tests for the system demonstrate that (i) the imaging depth is 28 mm, (ii) the optical background is sufficiently low and uniform, (iii) the non-uniform response of the optical imaging can be corrected by the flat field correction, and (iv) the imaging acquisition speed was improved by an average of 3.7 times faster than our previous systems. We also presented a geometry calibration procedure to map the planar BLIs acquired at multi-projections onto the surface of the CBCT image. The CBCT is required to generate the mesh for BLT reconstruction and used for treatment planning and radiation delivery. Feasibility study of the geometry calibration was performed on a manual-docking prototype. The mean and maximum mapping accuracy is 0.3 and 0.6 mm. The performance of the proposed motorized dual-use system is expected to be superior to that of the manual-docking prototype because of the mechanism stability. We anticipate the dual-use system as a highly efficient and cost-effective platform to facilitate optical imaging for preclinical radiation research.

Optical imaging has long been hindered by the high absorption and scattering of light in biological tissue. This makes it difficult to probe beyond a few millimeters beneath the surface without sacrificing image resolution and quantitative accuracy. Strong scattering and the inherent nature of the inverse problem makes fluorescence diffuse optical tomography (FT) extremely challenging. To this end, multi-modality techniques that combine anatomical imaging with the functional optical information have been used to improve the resolution and accuracy of FT. Previously, we have reported on the feasibility of a new imaging method, "Thermal Outlining using Focused Ultrasound" (TOFU), which combines the sensitivity of FT with the resolution of focused ultrasound using temperature reversible fluorescent probes. In this method, the position of the temperature reversible fluorescent probes is localized by an increase in fluorescent signal when the hot spot of the focused ultrasound beam is scanned over the medium. This a priori information is then utilized to guide and constrain conventional reconstruction algorithm to recover the position and concentration of the probes more accurately. The small size of the focal spot (~1.4 mm) up to a depth of 6 cm, allows imaging the distribution of these temperature sensitive agents with not only high spatial resolution but also high quantitative accuracy in deep tissue. In this work, the performance of the system will be evaluated using simulation and phantoms to investigate the dependence that size of the fluorescent distribution has on the TOFU system performance.

Imaging applications increasingly require light sources with high spectral density (power over spectral bandwidth. This has led in many cases to the replacement of conventional thermal light sources with bright light-emitting diodes (LEDs), lasers and superluminescent diodes. Although lasers and superluminescent diodes appear to be ideal light sources due to their narrow bandwidth and power, however, in the case of full-field imaging, their spatial coherence leads to coherent artefacts, such as speckle, that corrupt the image. LEDs, in contrast, have lower spatial coherence and thus seem the natural choice, but they have low spectral density. Random Lasers are an unconventional type of laser that can be engineered to provide low spatial coherence with high spectral density. These characteristics makes them potential sources for biological imaging applications where specific absorption and reflection are the characteristics required for state of the art imaging. In this work, a Random Laser (RL) is used to demonstrate speckle-free full-field imaging for polarization-dependent imaging in an epi-illumination configuration. We compare LED and RL illumination analysing the resulting images demonstrating that the RL illumination produces an imaging system with higher performance (image quality and spectral density) than that provided by LEDs.

Dynamic fluorescence molecular tomography (FMT) has the potential to quantify physiological or biochemical information, known as pharmacokinetic parameters, which are important for cancer detection, drug development and delivery etc. To image those parameters, there are indirect methods, which are easier to implement but tend to provide images with low signal-to-noise ratio, and direct methods, which model all the measurement noises together and are statistically more efficient. The direct reconstruction methods in dynamic FMT have attracted a lot of attention recently. However, the coupling of tomographic image reconstruction and nonlinearity of kinetic parameter estimation due to the compartment modeling has imposed a huge computational burden to the direct reconstruction of the kinetic parameters. In this paper, we propose to take advantage of both the direct and indirect reconstruction ideas through a variable splitting strategy under the augmented Lagrangian framework. Each iteration of the direct reconstruction is split into two steps: the dynamic FMT image reconstruction and the node-wise nonlinear least squares fitting of the pharmacokinetic parameter images. Through numerical simulation studies, we have found that the proposed algorithm can achieve good reconstruction results within a small amount of time. This will be the first step for a combined dynamic PET and FMT imaging in the future.

A pressure ulcer is one of the most important concerns for wheelchair bound patients with spinal cord injuries. A pressure ulcer is a localized injury near the buttocks that bear ischial tuberosity oppression over a long period of time. Due to elevated compression to blood vessels, the surrounding tissues suffer from a lack of oxygen and nutrition. The ulcers eventually lead to skin damage followed by tissue necrosis. The current medical strategy is to minimize the occurrence of pressure ulcers by regularly helping patients change their posture. However, these methods do not always work effectively or well. As a solution to fundamentally prevent pressure ulcers, a smart air cushion system was developed to detect and control pressure actively. The air cushion works by automatically adjusting a patient’s sitting posture to effectively relieve the buttock pressure.

To analyze the correlation between the dynamic pressure profiles of an air cell with a patient’s weight, a projection Moiré system was adopted to measure the deformation of an air cell and its associated stress distribution. Combining a full-field deformation imaging with air pressure measured within an air cell, the patient’s weight and the stress distribution can be simultaneously obtained. By integrating a full-field optical metrology with a time varying pressure sensor output coupled with different active air control algorithms for various designs, we can tailor the ratio of the air cells. Our preliminary data suggests that this newly developed smart air cushion has the potential to selectively reduce localized compression on the tissues at the buttocks. Furthermore, it can take a patient’s weight which is an additional benefit so that medical personnel can reference it to prescribe the correct drug dosages.

As a near-infrared (NIR) fluorescence dye, Indocyanine Green (ICG) has not gained broader clinical applications, owing to its multiple limitations such as concentration-dependent aggregation, low fluorescence quantum yield, poor physicochemical stability and rapid elimination from the body. In the meanwhile, 2H,3H-perfluoropentane (H-PFP) has been widely studied in ultrasound imaging as a vehicle for targeted delivery of contrast agents and drugs. We synthesized a novel dual-modal fluorescence and ultrasound contrast agent by encapsulating ICG and H-PFP in lipid microbubbles using a liquid-driven coaxial flow focusing (LDCFF) process. Uniform microbubbles with the sizes ranging from 1-10um and great ICG loading efficiency was achieved by this method. Our benchtop experiments showed that ICG/H-PFP microbubbles exhibited less aggregation, increased fluorescence intensity and more stable photostability compared to free ICG aqueous solution. Our phantom experiments demonstrated that ICG/H-PFP microbubbles enhanced the imaging contrasts in fluorescence imaging and ultrasonography. Our animal experiments indicated that ICG/H-PFP microbubbles extended the ICG life time and facilitated dual mode fluorescence and ultrasound imaging in vivo.

Tuberculosis is a highly contagious disease such that global latent patient can be as high as one third of the world population. Currently, latent tuberculosis was diagnosed by stimulating the T cells to produce the biomarker of tuberculosis, i.e., interferon-γ. In this paper, we developed a paraboloidal mirror enabled surface plasmon resonance (SPR) interferometer that has the potential to also integrate ellipsometry to analyze the antibody and antigen reactions. To examine the feasibility of developing a platform for cross calibrating the performance and detection limit of various bio-detection techniques, electrochemical impedance spectroscopy (EIS) method was also implemented onto a biochip that can be incorporated into this newly developed platform.

The microfluidic channel of the biochip was functionalized by coating the interferon-γ antibody so as to enhance the detection specificity. To facilitate the processing steps needed for using the biochip to detect various antigen of vastly different concentrations, a kinetic mount was also developed to guarantee the biochip re-positioning accuracy whenever the biochip was removed and placed back for another round of detection. With EIS being utilized, SPR was also adopted to observe the real-time signals on the computer in order to analyze the success of each biochip processing steps such as functionalization, wash, etc. Finally, the EIS results and the optical signals obtained from the newly developed optical detection platform was cross-calibrated. Preliminary experimental results demonstrate the accuracy and performance of SPR and EIS measurement done at the newly integrated platform.

The optimization of experimental design prior to deployment, not only for cost effective solution but also for computationally efficient image reconstruction has taken up for this study. We implemented the iterative method also known as effective independence (EFI) method for optimization of source/detector pair configuration. The notion behind for adaptive selection of minimally correlated measurements was to evaluate the information content passed by each measurement for estimation of unknown parameter. The EFI method actually ranks measurements according to their contribution to the linear independence of unknown parameter basis. Typically, to improve the solvability of ill conditioned system, regularization parameter is added, which may affect the source/detector selection configuration. We show that the source/detector pairs selected by EFI method were least prone to vary with sub optimal regularization value. Moreover, through series of simulation studies we also confirmed that sparse source/detector pair measurements selected by EFI method offered similar results in comparison with the dense measurement configuration for unknown parameters qualitatively as well as quantitatively. Additionally, EFI method also allow us to incorporate the prior knowledge, extracted in multimodality imaging cases, to design source/detector configuration sensitive to specific region of interest. The source/detector ranking method was further analyzed to derive the automatic cut off number for iterative scheme.

This proceedings shows the combination of Optical Coherence Tomography (OCT) and Hyper-Spectral Imaging (HSI) using a double-clad optical fiber. The single mode core of the fiber is used to transmit OCT signals, while the cladding, with its large collection area, provides an efficient way to capture the reflectance spectrum of the sample. The combination of both methods enables three-dimensional acquisition of sample morphology with OCT, enhanced by the molecular information contained in its hyper-spectral image. We believe that the combination of these techniques could result in endoscopes with enhanced tissue identification capability.

The Spatial-frequency dependence of turbid media reflectance encodes both optical properties and depth information. The high spatial frequency domain imaging (HSFDI) can, in particular, extract key characteristics of the phase function of the scattering medium which carries the ultimate structural information of the medium. We first outline the principle of HSFDI and then present here a compact optical configuration integrating the modulated illumination and imaging systems, facilitating quantitative wide-field optical properties mapping at high spatial frequencies. The performance of HSFDI is assessed on both tissue phantoms and in vivo.

Traditional photoplethysmographic imaging (PPGI) systems use the red, green, and blue (RGB) broadband measurements of a consumer digital camera to remotely estimate a patients heart rate; however, these broadband RGB signals are often corrupted by ambient noise, making the extraction of subtle fluctuations indicative of heart rate difficult. Therefore, the use of narrow-band spectral measurements can significantly improve the accuracy. We propose a novel digital spectral demultiplexing (DSD) method to infer narrow-band spectral information from acquired broadband RGB measurements in order to estimate heart rate via the computation of motion- compensated skin erythema fluctuation. Using high-resolution video recordings of human participants, multiple measurement locations are automatically identified on the cheeks of an individual, and motion-compensated broadband reflectance measurements are acquired at each measurement location over time via measurement location tracking. The motion-compensated broadband reflectance measurements are spectrally demultiplexed using a non-linear inverse model based on the spectral sensitivity of the camera's detector. A PPG signal is then computed from the demultiplexed narrow-band spectral information via skin erythema fluctuation analysis, with improved signal-to-noise ratio allowing for reliable remote heart rate measurements. To assess the effectiveness of the proposed system, a set of experiments involving human motion in a front-facing position were performed under ambient lighting conditions. Experimental results indicate that the proposed system achieves robust and accurate heart rate measurements and can provide additional information about the participant beyond the capabilities of traditional PPGI methods.

Impact trauma may cause a hematoma, which is the leakage of venous blood into surrounding tissues. Large hematomas can be dangerous as they may inhibit local blood ow. Hematomas are often diagnosed visually, which may be problematic if the hematoma leaks deeper than the visible penetration depth. Furthermore, vascular wound healing is often monitored at home without the aid of a clinician. We therefore investigated the use of near infrared (NIR) re ectance photoplethysmographic imaging (PPGI) to assess vascular damage resulting from a hematoma, and monitor the healing process. In this case study, the participant experienced internal vascular damage in the form of a hematoma. Using a PPGI system with dual-mode temporally coded illumination for ambient-agnostic data acquisition and mounted optical elements, the tissue was illuminated with a spatially uniform irradiance pattern of 850 nm wavelength light for increased tissue penetration and high oxy-to-deoxyhemoglobin absorption ratio. Initial and follow-up PPGI data collection was performed to assess vascular damage and healing. The tissue PPGI sequences were spectrally analyzed, producing spectral maps of the tissue area. Experimental results show that spatial differences in spectral information can be observed around the damaged area. In particular, the damaged site exhibited lower pulsatility than the surrounding healthy tissue. This pulsatility was largely restored in the follow-up data, suggesting that the tissue had undergone vascular healing. These results indicate that hematomas can be assessed and monitored in a non-contact visual manner, and suggests that PPGI can be used for tissue health assessment, with potential extensions to peripheral vascular disease.

Continuous heart rate monitoring can provide important context for quantitative clinical assessment in scenarios such as long-term health monitoring and disability prevention. Photoplethysmographic imaging (PPGI) systems are particularly useful for such monitoring scenarios as contact-based devices pose problems related to comfort and mobility. Each pixel can be regarded as a virtual PPG sensor, thus enabling simultaneous measurements of multiple skin sites. Existing PPGI systems analyze temporal PPGI sensor uctuations related to hemodynamic pulsations across a region of interest to extract the blood pulse signal. However, due to spatially varying optical properties of the skin, the blood pulse signal may not be consistent across all PPGI sensors, leading to inaccurate heart rate monitoring. To increase the hemodynamic signal-to-noise ratio (SNR), we propose a novel spectral PPGI sensor fusion method for enhanced estimation of the true blood pulse signal. Motivated by the observation that PPGI sensors with high hemodynamic SNR exhibit a spectral energy peak at the heart rate frequency, an entropy-based fusion model was formulated to combine PPGI sensors based on the sensors' spectral energy distribution. The optical PPGI device comprised a near infrared (NIR) sensitive camera and an 850 nm LED. Spatially uniform irradiance was achieved by placing optical elements along the LED beam, providing consistent illumination across the skin area. Dual-mode temporally coded illumination was used to negate the temporal effect of ambient illumination. Experimental results show that the spectrally weighted PPGI method can accurately and consistently extract heart rate information where traditional region-based averaging fails.

Single-pixel imaging based on compressive sensing theory has been a highlighted technique in the biomedical imaging field for many years. This interest has been driven by the possibility of performing microscopic or macroscopic imaging based on low-cost detector arrays, increased SNR (signal-to-noise ratio) in the acquired data sets and the ability to perform high quality image reconstruction with compressed data sets by exploiting signal sparsity. In this work, we present our recent work in implementing this technique to perform time domain fluorescence-labeled investigations in preclinical settings. More precisely, we report on our time-resolved hyperspectral single-pixel camera for fast, wide-field mapping of molecular labels and lifetime-based quantification. The hyperspectral single-pixel camera implements a DMD (Digital micro-mirror device) to generate optical masks for modulating the illumination field before it is delivered onto the sample and focuses the emission light signals into a multi-anode hyperspectral time-resolved PMT (Photomultiplier tube) to acquire spatial, temporal and spectral information enriched 4-D data sets. Fluorescence dyes with lifetime and spectral contrast are embedded in well plates and thin tissues. L-1 norm based regularization or the least square method, is applied to solve the underdetermined inverse problem during image reconstruction. These experimental results prove the possibility of fast, wide-field mapping of fluorescent labels with lifetime and spectral contrast in thin media.

Fluorescence Molecular Tomography (FMT) is a powerful optical imaging tool for preclinical research. Especially, its implementation with time-domain (TD) techniques allows lifetime multiplexing for simultaneously imaging multiple biomarkers and provides enhanced data sets for improved resolution and quantification compared to continuous wave (CW) and frequency domain (FD) methodologies. When performing time-domain reconstructions, one important aspect is the selection of a temporal sub-data set. Typically, such selection is performed a posteriori after dense temporal sampling during the acquisition. In this work, we investigate the potential to collect a priori sparse data sets for fast experimental acquisition without compromising FMT performances.

Förster Resonance Energy Transfer (FRET) is widely used to sense molecular interactions occurring at the nanoscale. In vitro and ex vivo protocols for visualizing FRET are already well-established, but in vivo studies have proven to be more challenging. One issue that hinders in vivo visualization of FRET is the higher absorption and scattering of visible light within tissues. In this case, light in the near-infrared (NIR) spectral window is required for increased depth sensing. Moreover, due to spectral variation in optical properties as well as heterogeneous spatial distribution, lifetime-based FRET imaging is preferred. Herein, we investigate the effect of temporal acquisition settings on the lifetime-based estimation of the fraction of quenched donor molecules (A1) as well as the quenched donor lifetime (τ₁). We performed in silico, in vitro, and in vivo experiments under gate widths of 300ps to 1000ps in 100ps intervals to determine the effect on quantification of A₁ and τ₁. Even though the NIR fluorescent dyes have shorter lifetimes then visible fluorophores, we were still able to accurately quantify FRET under all tested system gate widths and experimental conditions.