This PDF file contains the front matter associated with SPIE Proceedings Volume 8935, including the Title Page, Copyright information, Table of Contents, Invited Panel Discussion, and Conference Committee listing.

The principles of cancer treatment has for years been radical resection of the primary tumor. In the oncologic surgeries where the affected cancer site is close to the lymphatic system, it is as important to detect the draining lymph nodes for metastasis (lymph node mapping). As a replacement for conventional radioactive labeling, indocyanine green (ICG) has shown successful results in lymph node mapping; however, most of the ICG fluorescence detection techniques developed are based on camera imaging. In this work, fluorescence spectroscopy using a fiber-optical probe was evaluated on a tissue-like ICG phantom with ICG concentrations of 6-64 μM and on breast tissue from five patients. Fiber-optical based spectroscopy was able to detect ICG fluorescence at low intensities; therefore, it is expected to increase the detection threshold of the conventional imaging systems when used intraoperatively. The probe allows spectral characterization of the fluorescence and navigation in the tissue as opposed to camera imaging which is limited to the view on the surface of the tissue.

Successful diagnosis, screening, and elimination of malaria critically depend on rapid and sensitive detection of this dangerous infection, preferably transdermally and without sophisticated reagents or blood drawing. Such diagnostic methods are not currently available. Here we show that the high optical absorbance and nanosize of endogenous heme nanoparticles called hemozoin, a unique component of all blood-stage malaria parasites, generate a transient vapor nanobubble around hemozoin in response to a short and safe near-infrared picosecond laser pulse. The acoustic signals of these malaria-specific nanobubbles provided the first transdermal non-invasive and rapid detection of a malaria infection as low as 0.00034% in animals without using any reagents or drawing blood. These on-demand transient events have no analogs among current malaria markers and probes, can detect and screen malaria in seconds and can be realized as a compact, easy to use, inexpensive and safe field technology.

Multi-functional laser non-invasive diagnostic systems, such as “LAKK-M”, allow the study of a number of microcirculatory parameters, including blood microcirculatory index (I_m) (by laser Doppler flowmetry, LDF) and oxygen saturation (S_tO₂) of skin tissue (by tissue reflectance oximetry, TRO). Such systems may provide significant information relevant to physiology and clinical medicine. The aim of this research was to use such a system to study the synchronization of microvascular blood flow and oxygen saturation rhythms under normal and adaptive change conditions. Studies were conducted with 8 healthy volunteers – 3 females and 5 males of 21-49 years. Each volunteer was subjected to basic 3 minute tests. The volunteers were observed for between 1-4 months each, totalling 422 basic tests. Measurements were performed on the palmar surface of the right middle finger and the forearm medial surface. Wavelet analysis was used to study rhythmic oscillations in LDF- and TRO-data. Tissue oxygen consumption (from arterial and venal blood oxygen saturation and nutritive flux volume) was calculated for all volunteers during “adaptive changes” as (617±123 AU) and (102±38 AU) with and without arteriovenous anastomoses (AVAs) respectively. This demonstrates increased consumption compared to normal (495±170 AU) and (69±40 AU) with and without AVAs respectively. Data analysis demonstrated the emergence of resonance and synchronization of rhythms of microvascular blood flow and oxygen saturation as an adaptive change in myogenic oscillation (vasomotion) resulting from exercise and potentially from psychoemotional stress. Synchronization of myogenic rhythms during adaptive changes suggest increased oxygen consumption resulting from increased microvascular blood flow velocity.

There has been a growing interest in the development of a low cost depth-resolved non-invasive dermis imaging tool for both clinical and fundamental investigations of skin diseases. Multiple reference optical coherence tomography (MR-OCT) is a recently developed miniature time-domain low coherence interferometeric imaging platform, which promises to fit into robust, cost-effective designs that are virtually solid state, typical of handheld devices. In this paper we demonstrate the feasibility of MR-OCT for dermis imaging applications by incorporating it in a dermascope, which provides simultaneous imaging of dermis and an interactive tool for beam steering and registration of the OCT imaging beam at the dermis area. This allows the user to interactively investigate the depth resolved information of any target position of interest on the dermis by pointing the mouse cursor within the dermis image or selecting the area on a touch screen. Image acquisition is controlled with software that displays both the dermis and MR-OCT axial-scan, and allows detailed information of the depth scan signal to screen for skin disease. We believe this approach will have a significant impact on medical care.

Nanotechnology provides tremendous biomedical opportunities for cancer diagnosis, imaging, and therapy. In contrast to conventional chemotherapeutic agents where their actual target delivery cannot be easily imaged, integrating imaging and therapeutic properties into one platform facilitates the understanding of pharmacokinetic profiles, and enables monitoring of the therapeutic process in each individual. Such a concept dubbed “theranostics” potentiates translational research and improves precision medicine. One particular challenging application of theranostics involves imaging and controlled delivery of nanoplatforms across blood-brain-barrier (BBB) into brain tissues. Typically, the BBB hinders paracellular flux of drug molecules into brain parenchyma. BBB disrupting agents (e.g. mannitol, focused ultrasound), however, suffer from poor spatial confinement. It has been a challenge to design a nanoplatform not only acts as a contrast agent but also improves the BBB permeation. In this study, we demonstrated the feasibility of plasmonic gold nanoparticles as both high-resolution optical contrast agent and focalized tumor BBB permeation-inducing agent. We specifically examined the microscopic distribution of nanoparticles in tumor brain animal models. We observed that most nanoparticles accumulated at the tumor periphery or perivascular spaces. Nanoparticles were present in both endothelial cells and interstitial matrices. This study also demonstrated a novel photothermal-induced BBB permeation. Fine-tuning the irradiating energy induced gentle disruption of the vascular integrity, causing short-term extravasation of nanomaterials but without hemorrhage. We conclude that our gold nanoparticles are a powerful biocompatible contrast agent capable of inducing focal BBB permeation, and therefore envision a strong potential of plasmonic gold nanoparticle in future brain tumor imaging and therapy.

Near-infrared fluorescence image-guided surgery, FIGS, has lately shown a huge potential in oncologic and lymphatic related surgeries. In some indications such as liver or heart surgery, fluorescence-reachable anatomic structures are limited by the access to the surgical field. Nevertheless, most of the systems available on the market are too large to image the sides of cavities. Small devices are clearly required to improve workability of fluorescence imaging systems. The current work describes the development of an instrument and the results of its evaluation. In order to image narrow area, we developed a small size device consisting of an optical head connected to a control box. The whole system, optical head, control box and software, receives a CE mark for clinical procedures. Building on existing technologies, we simplified the fluorescence imaging system. It consists of a custom charged-coupled device camera, a high color rendering index visible LED illumination and a Class1 Laser fluorophore excitation. With a curved shape of 25x35x150mm, the optical head was designed as a true hand-held probe. The field of view varies from 5x3.75cm to 2x1.5cm. The device is able to collect and display the signal of 5pmol of IndoCyanine Green (ICG) with a spatial resolution down to 70μm at 25 frames per second. The system has been evaluated in pre-clinical and clinical procedures. The preclinical studies confirmed the ability of the system to visualize tumors in mice models. Clinical evaluations includes lymphedema investigations and surgical resections of tumors in colorectal cancer.

We describe the development and performance analysis of two clinical near-infrared fluorescence image-guided surgery (FIGS) devices that aim to overcome some of the limitations of current FIGS systems. The devices operate in a widefield-imaging mode and can work (1) in conjunction with a laparoscope, during minimally invasive surgery, and (2) as a hand-held, open surgery imaging system. In both cases, narrow-band excitation light, delivered at multiple wavelengths, is efficiently combined with white reflectance light. Light is delivered to ~100 cm² surgical field at 1-2 mW/cm² for white light and 3-7 mW/cm² (depending on wavelength) of red - near infrared excitation, at a typical working distance of 350 mm for the hand-held device and 100 mm for the laparoscope. A single, sensitive, miniaturized color camera collects both fluorescence and white reflectance light. The use of a single imager eliminates image alignment and software overlay complexity. A novel filtering and illumination arrangement allows simultaneous detection of white reflectance and fluorescence emission from multiple dyes in real-time. We will present both fluorescence detection sensitivity modeling and practical performance data. We have demonstrated the efficiency and the advantages of the devices both pre-clinically and during live surgery on humans. Both the hand-held and the laparoscopic systems have proved to be reliable and beneficial in an ongoing clinical trial involving sentinel lymph node detection in gynecological cancers. We will show preliminary results using two clinically approved dyes, Methylene blue and indocyanine green. We anticipate that this technology can be integrated and routinely used in a larger variety of surgical procedures.

Obtaining three-dimensional (3D) information of biologic tissue is important in many medical applications. This paper presents two methods for reconstructing 3D topography of biologic tissue: multiview imaging and structured light illumination. For each method, the working principle is introduced, followed by experimental validation on a diabetic foot model. To compare the performance characteristics of these two imaging methods, a coordinate measuring machine (CMM) is used as a standard control. The wound surface topography of the diabetic foot model is measured by multiview imaging and structured light illumination methods respectively and compared with the CMM measurements. The comparison results show that the structured light illumination method is a promising technique for 3D topographic imaging of biologic tissue.

The wound healing process involves the reparative phases of inflammation, proliferation, and remodeling. Interrupting any of these phases may result in chronically unhealed wounds, amputation, or even patient death. Despite the clinical significance in chronic wound management, no effective methods have been developed for quantitative image-guided treatment. We integrated a multimodal imaging system with a cold atmospheric plasma probe for image-guided treatment of chronic wound. Multimodal imaging system offers a non-invasive, painless, simultaneous and quantitative assessment of cutaneous wound healing. Cold atmospheric plasma accelerates the wound healing process through many mechanisms including decontamination, coagulation and stimulation of the wound healing. The therapeutic effect of cold atmospheric plasma is studied in vivo under the guidance of a multimodal imaging system. Cutaneous wounds are created on the dorsal skin of the nude mice. During the healing process, the sample wound is treated by cold atmospheric plasma at different controlled dosage, while the control wound is healed naturally. The multimodal imaging system integrating a multispectral imaging module and a laser speckle imaging module is used to collect the information of cutaneous tissue oxygenation (i.e. oxygen saturation, StO₂) and blood perfusion simultaneously to assess and guide the plasma therapy. Our preliminary tests show that cold atmospheric plasma in combination with multimodal imaging guidance has the potential to facilitate the healing of chronic wounds.

We describe a portable fluorescence goggle navigation system for cancer margin assessment during oncologic surgeries. The system consists of a computer, a head mount display (HMD) device, a near infrared (NIR) CCD camera, a miniature CMOS camera, and a 780 nm laser diode excitation light source. The fluorescence and the background images of the surgical scene are acquired by the CCD camera and the CMOS camera respectively, co-registered, and displayed on the HMD device in real-time. The spatial resolution and the co-registration deviation of the goggle navigation system are evaluated quantitatively. The technical feasibility of the proposed goggle system is tested in an ex vivo tumor model. Our experiments demonstrate the feasibility of using a goggle navigation system for intraoperative margin detection and surgical guidance.

Although fs-laser surgery is clinically established in the field of corneal flap cutting for laser in situ keratomileusis, surgery with fs-laser in the posterior part of the eye is impaired by focus degradation due to aberrations. Precise targeting and keeping of safety distance to the retina also relies on an intraoperative depth resolved imaging. We demonstrate a concept for image guided fs-laser surgery in the vitreous body combining adaptive optics (AO) for focus reshaping and optical coherence tomography (OCT) for focus position guidance. The setup of the laboratory system consist of an 800 nm fs-laser which is focused into a simple eye model via a closed loop adaptive optics system with Hartmann-Shack sensor and a deformable mirror to correct for wavefront aberrations. A spectral domain optical coherence tomography system is used to target phantom structures in the eye model. Both systems are set up to share the same scanner and focusing optics. The use of adaptive optics results in a lowered threshold energy for laser induced breakdown and an increased cutting precision. 3D OCT imaging of porcine retinal tissue prior and immediately after fs-laser cutting is also demonstrated. In the near future OCT and AO will be two essential assistive components in possible clinical systems for fs-laser based eye surgery beyond the cornea.

Shifted excitation Raman difference spectroscopy (SERDS) using a dual-wavelength laser diode laser as excitation light source at 671 nm is presented. This device has a size of 3 mm x 0.5 mm and contains two laser cavities with wavelengths adjusted by distributed Bragg reflector (DBR) gratings as rear side mirrors. An integrated Y-branch coupler guides the emission into a common output aperture. The two wavelengths are centered at 671 nm with a well-defined spectral spacing of about 10 cm^-1. An output power up to 100 mW is achieved. Raman experiments using polystyrene as test sample and ambient light to disturb the Raman signals demonstrate the suitability of such light source for SERDS.

We present the use of a commercially available electrically tunable lens to achieve axial scanning in a reflectance confocal microscope. Over a 255 μm axial scan range, the lateral and axial resolutions varied from 1-2 μm and 4-14 μm, respectively, dependent on the variable focal length of the tunable lens. Confocal imaging was performed on normal human biopsies from the oral cavity ex vivo. Sub-cellular morphologic features were seen throughout the depth of the epithelium while axially scanning using the focus tunable lens.

Polarization measurements allow one to enhance imaging contrast of superficial tissues and obtain new polarization sensitive parameters for better description of the micro- and macro- structural and optical properties of complex tissues. Since the majority of cancers originate in the epithelial layer, probing the morphological and pathological changes in the superficial tissues using an expended parameter set and with improved spatial resolution and contrasts will lead to new clues on the early clinical detection of cancers. In this wok, we carry out polarization imaging on cancerous tissues and look for cancer specific features. Using a scattering model, which approximates the anisotropic biological tissues to a mixture of spherical and cylindrical scatterers imbedded in birefringent ambient media, and a Monte Carlo simulation program, we examine the relationship between the micro-structure of the model and the characteristic polarization features. The studies help to understand the contrast mechanism of polarization sensitive measurements for different cancers and provide the basis for potential clinical applications.

Long period fiber gratings (LPFGs) have been proposed as label-free optical biosensor for a few years. Refractive index changes, which modify the fiber transmission spectrum, are still used for evaluating a biochemical interaction that occurs along the grating region. A turn-around point (TAP) LPFG was manufactured for enhancing the refractive index sensitivity of these devices. Considering the simplicity and the fast process with respect to the silanization procedure, the functionalization of the fiber was carried out by Eudragit L100 copolymer. An IgG/anti-IgG immunoassay was implemented for studying the antigen/antibody interaction. A limit of detection lower than 100 μg L^-1 was achieved. Based on the same model assay, we compared the resonance wavelength shifts during the injection of 10 mg L^-1 anti-IgG antigen between the TAP LPFG and a standard non-TAP one, in which the coupling occurs with a lower order cladding mode, as performance improvement of the LPFG-based biosensors.

Currently, it has been an international focus on intraoperative precise positioning and accurate resection of tumor and metastases. The methods such as X-rays, computed tomography (CT), magnetic resonance imaging (MRI) and positron emission tomography (PET) have played an important role in preoperative accurate diagnosis. However, most of them are inapplicable for intraoperative surgery. We have proposed a surgical navigation system based on optical molecular imaging technology for intraoperative detection of tumors and metastasis. This system collects images from two CCD cameras for real-time fluorescent and color imaging. For image processing, the template matching algorithm is used for multispectral image fusion. For the application of tumor detection, the mouse breast cancer cell line 4T1-luc, which shows highly metastasis, was used for tumor model establishment and a model of matrix metalloproteinase (MMP) expressing breast cancer. The tumor–bearing nude mice were given tail vein injection of MMP 750FAST (PerkinElmer, Inc. USA) probe and imaged with both bioluminescence and fluorescence to assess in vivo binding of the probe to the tumor and metastases sites. Hematoxylin and eosin (H&E) staining was performed to confirm the presence of tumor and metastasis. As a result, one tumor can be observed visually in vivo. However liver metastasis has been detected under surgical navigation system and all were confirmed by histology. This approach helps surgeons to find orthotopic tumors and metastasis during intraoperative resection and visualize tumor borders for precise positioning. Further investigation is needed for future application in clinics.

Quanterix and Stratec Biomedical have developed an instrument that enables the automated measurement of multiple proteins at concentration ~1000 times lower than existing immunoassays. The instrument is based on Quanterix’s proprietary Single Molecule Array technology (Simoa™ ) that facilitates the detection and quantification of biomarkers previously difficult to measure, thus opening up new applications in life science research and in-vitro diagnostics. Simoa is based on trapping individual beads in arrays of femtoliter-sized wells that, when imaged with sufficient resolution, allows for counting of single molecules associated with each bead. When used to capture and detect proteins, this approach is known as digital ELISA (Enzyme-linked immunosorbent assay). The platform developed is a merger of many science and engineering disciplines. This paper concentrates on the optical technologies that have enabled the development of a fully-automated single molecule analyzer. At the core of the system is a custom, wide field-of-view, fluorescence microscope that images arrays of microwells containing single molecules bound to magnetic beads. A consumable disc containing 24 microstructure arrays was developed previously in collaboration with Sony DADC. The system cadence requirements, array dimensions, and requirement to detect single molecules presented significant optical challenges. Specifically, the wide field-of-view needed to image the entire array resulted in the need for a custom objective lens. Additionally, cost considerations for the system required a custom solution that leveraged the image processing capabilities. This paper will discuss the design considerations and resultant optical architecture that has enabled the development of an automated digital ELISA platform.

Intraoperative fluorescence imaging in reflectance geometry (FRI) is an attractive imaging modality as it allows to noninvasively monitor the fluorescence targeted tumors located below the tissue surface. Some drawbacks of this technique are the background fluorescence decreasing the contrast and absorption heterogeneities leading to misinterpretations concerning fluorescence concentrations. We presented a FRI technique relying on a laser line scanning instead of a uniform illumination. Here, we propose a correction technique based on this illumination scheme. We scan the medium with the laser line and acquire at each position of the line both fluorescence and excitation images. We then use the finding that there is a relationship between the excitation intensity profile and the background fluorescence one. This allows us to predict the amount of signal to subtract to the fluorescence images to get a better contrast. As the light absorption information is contained both in fluorescence and excitation images, this method also permits us to correct the effects of absorption heterogeneities, leading to a better accuracy for the detection. This technique has been validated on simulations (with a Monte-Carlo code and with the diffusion approxi- mation using NIRFAST) and experimentally with tissue-like liquid phantoms with different levels of background fluorescence. Fluorescent inclusions are observed in several configurations at depths ranging from 1 mm to 1 cm. Results obtained with this technique are compared to those obtained with a more classical wide-field detection scheme for the contrast enhancement and to the fluorescence to excitation ratio approach for the absorption correction.

Uterine transplantation surgery has been proposed as a treatment for permanent absolute uterine factor infertility (AUFI) in the case of loss of the uterus. Due to the complexity of the vasculature correct reanastomosis of the blood supply during transplantation surgery is a crucial step to ensure reperfusion and viability of the organ. While techniques such as fluorescent dye imaging have been proposed to visualise perfusion there is no gold standard for intraoperative visualisation of tissue oxygenation. In this paper results from a liquid crystal tuneable filter (LCTF)-based multispectral imaging (MSI) laparoscope are described. The system was used to monitor uterine oxygen saturation (SaO₂) before and after transplantation. Results from surgeries on two animal models (rabbits and sheep) are presented. A feature-based registration algorithm was used to correct for misalignment induced by breathing or peristalsis in the tissues of interest prior to analysis. An absorption spectrum was calculated at each spatial pixel location using reflectance data from a reference standard, and the relative contributions from oxy- and deoxyhaemoglobin were calculated using a least squares regression algorithm with non-negativity constraints. Results acquired during animal surgeries show that cornual oxygenation changes are consistent with those observed in point measurements taken using a pulse oximeter, showing reduced SaO₂ following reanastomosis. Values obtained using the MSI laparoscope were lower than those taken with the pulse oximeter, which may be due to the latter’s use of the pulsatile arterial blood signal. Future work incorporating immunological test results will help to correlate SaO₂ levels with surgical outcomes.

Primary care physicians must conduct a staggering number of comprehensive physical exams and medical record reviews, resulting in demanding daily schedules. Few commercial technologies have been marketed towards the primary care market, which has stifled improvements in disease screening and detection, work flow, and records management, taking time away from interactions with patients. In efforts to improve the quality of care in primary care medicine, we integrated our handheld primary care optical imaging system with Google Glass©, a commercial heads-up display (HUD). The integration of a HUD allows the physician to focus on the patient during the medical history review and during the patient exam, resulting in potential improvements to the quality of care and efficient access to real-time data for display and analysis.

A system has been developed to assess the feasibility of using motion tracking to enable pre-surgical margin mapping of basal cell carcinoma (BCC) in the clinic using optical coherence tomography (OCT). This system consists of a commercial OCT imaging system (the VivoSight 1500, MDL Ltd., Orpington, UK), which has been adapted to incorporate a webcam and a single-sensor electromagnetic positional tracking module (the Flock of Birds, Ascension Technology Corp, Vermont, USA). A supporting software interface has also been developed which allows positional data to be captured and projected onto a 2D dermoscopic image in real-time. Initial results using a stationary test phantom are encouraging, with maximum errors in the projected map in the order of 1-2mm. Initial clinical results were poor due to motion artefact, despite attempts to stabilise the patient. However, the authors present several suggested modifications that are expected to reduce the effects of motion artefact and improve the overall accuracy and clinical usability of the system.

We report an improved method to visualize lipid distribution in axial and lateral direction within arterial vessel walls by spectroscopic spectral-domain Optical Coherence Tomography (OCT) at 1.7μm wavelength for identification of lipidrich plaque that is suspected to cause coronary events. In our previous method, an extended InGaAs-based line camera detects an OCT interferometric spectrum from 1607 to 1766 nm, which is then divided into twenty subbands, and A-scan OCT profile is calculated for each subband, resulting in a tomographic spectrum. This tomographic spectrum is decomposed into lipid spectrum having an attenuation peak at 1730 nm and non-lipid spectrum independent of wavelength, and the weight of each spectrum, that is, lipid and non-lipid score is calculated. In this paper, we present an improved algorithm, in which we have combined the lipid score and the non-lipid score to derive a corrected lipid score. We have found that the corrected lipid score is better than the raw lipid score in that the former is more robust against false positive occurring due to abrupt change in reflectivity at vessel surface. In addition, we have optimized spatial smoothing filter and reduced false positive and false negative due to detection noise and speckle. We have verified this improved algorithm by the use of measuring data of normal porcine coronary artery and lard as a model of lipid-rich plaque and confirmed that both the sensitivity and the specificity of lard are 92%.

Anastomosis is one of the most commonly performed procedure in the clinical environment that involves tubular structures, such as blood vessel, lymphatic vessel, seminal duct and ureter. Suture based anastomosis is still the foundation for most basic surgical training and clinical operation, although alternate techniques have been developed and under development. For those tubular-structure-anastomosis, immediate real-time post-operative evaluation of the surgical outcome is critical to the success of surgery. Previously evaluation is mostly based on surgeons' experience. Fourier-domain optical coherence tomography is high-speed, high-resolution noninvasive 3D imaging modality that has been widely used in the biomedical research and clinical study. In this study we used Fourier-domain optical coherence tomography as an evaluation tool for anastomosis of lymphatic vessels, ureter and seminal duct in rodent model. Immediate post-operative and long term surgical site data were collected and analyzed. Critical clinical parameters such as lumen patency, anastomosed site narrowing and suture error detection are provided to surgeons.

Breast-conserving surgery is a frequent option for women with stage I and II breast cancer, and with radiation treatment, can be as effective as a mastectomy. However, adequate margin detection remains a challenge, and too often additional surgeries are required. Optical coherence tomography (OCT) provides a potential method for real-time, high-resolution imaging of breast tissue during surgery. Intra-operative OCT imaging of excised breast tissues has been previously demonstrated by several groups. In this study, a novel handheld surgical probe-based OCT system is introduced, which was used by the surgeon to image in vivo, within the tumor cavity, and immediately following tumor removal in order to detect the presence of any remaining cancer. Following resection, study investigators imaged the excised tissue with the same probe for comparison. We present OCT images obtained from over 15 patients during lumpectomy and mastectomy surgeries. Images were compared to post-operative histopathology for diagnosis. OCT images with micron scale resolution show areas of heterogeneity and disorganized features indicative of malignancy, compared to more uniform regions of normal tissue. Video-rate acquisition shows the inside of the tumor cavity as the surgeon sweeps the probe along the walls of the surgical cavity. This demonstrates the potential of OCT for real-time assessment of surgical tumor margins and for reducing the unacceptably high re-operation rate for breast cancer patients.

Endoscope cameras play an important and growing role as a diagnostic and surgical tool. The endoscope camera is usually used to provide a view of the scene straight ahead of the instrument to the operator. As is common in many remotely operated systems, the limited field of view and the inability to pan the camera make it challenging to gain a situational awareness comparable to an operator with direct access to the scene. We present a spectral multiplexing technique for endoscopes that allows for overlay of the existing forward view with additional views at different angles to increase the effective field of view of the device. Our goal is to provide peripheral vision while minimally affecting the design and forward image quality of existing systems.

Photometric stereo endoscopy is a technique that captures information about the high-spatial-frequency topography of the field of view simultaneously with a conventional color image. Here we describe a system that will enable photometric stereo endoscopy to be clinically evaluated in the large intestine of human patients. The clinical photometric stereo endoscopy system consists of a commercial gastroscope, a commercial video processor, an image capturing and processing unit, custom synchronization electronics, white light LEDs, a set of four fibers with diffusing tips, and an alignment cap. The custom pieces that come into contact with the patient are composed of biocompatible materials that can be sterilized before use. The components can then be assembled in the endoscopy suite before use. The resulting endoscope has the same outer diameter as a conventional colonoscope (14 mm), plugs into a commercial video processor, captures topography and color images at 15 Hz, and displays the conventional color image to the gastroenterologist in real-time. We show that this system can capture a color and topographical video in a tubular colon phantom, demonstrating robustness to complex geometries and motion. The reported system is suitable for in vivo evaluation of photometric stereo endoscopy in the human large intestine.

This paper described evaluation of the three-dimensional endoscope system for assessing the gastrointestinal motility. Gastrointestinal diseases are mainly based on the morphological or anatomical abnormity. However, sometimes the gastrointestinal symptoms are apparent without visible abnormalities. Such diseases are called functional gastrointestinal disorder, for example, functional dyspepsia, and irritable bowel syndrome. One of the major factors of these diseases is the gastrointestinal dysmotility. Assessment procedures for motor function are either invasive, or indirect. We thus propose a three-dimensional endoscope system for assessing the gastrointestinal motility. To assess the dynamic motility of the stomach, three-dimensional endoscopic imaging of stomach lining is performed. Propagating contraction waves are detected by subtracting estimated stomach geometry without contraction waves from one with contraction waves. After detecting constriction waves, their frequency, amplitude, and speed of propagation can be calculated. In this study, we evaluate the proposed system. First, we evaluate the developed three-dimensional endoscope system by a flat plane. This system can measure the geometry of the flat plane with an error of less than 10 percent of the distance between endoscope tip and the object. Then we confirm the validity of a prototype system by a wave simulated model. The detected wave is approximated by a Gaussian function. In the experiment, the amplitude and position of the wave can be measure with 1 mm accuracy. These results suggest that the proposed system can measure the speed and amplitude of contraction. In the future, we evaluate the proposed system in vivo experiments.

Raman spectroscopy has proven to be a powerful tool for discriminating between normal and abnormal tissue types. Fiber based Raman probes have demonstrated its potential for in vivo disease diagnostics. Combining Raman spectroscopy with Magnetic Resonance Imaging (MRI) opens up new avenues for MR guided minimally invasive optical biopsy. Although Raman probes are commercially available, they are not compatible with a MRI environment due to the metallic components which are used to align the micro-optic components such as filters and lenses at the probe head. Additionally they are not mechanically compatible with a typical surgical environment as factors such as sterility and length of the probe are not addressed in those designs. We have developed an MRI compatible fiber Raman probe with a disposable probe head hence maintaining sterility. The probe head was specially designed to avoid any material that would cause MR imaging artefacts. The probe head that goes into patient’s body had a diameter <1.5 mm so that it is compatible with biopsy needles and catheters. The probe has been tested in MR environment and has been proven to be capable of obtaining Raman signal while the probe is under real-time MR guidance.

The ability to determine the depth and degree of cutaneous and subcutaneous tissue damage is critical for medical applications such as burns and pressure ulcers. The Diffuse Photon Density Wave (DPDW) methodology at near infrared wavelengths can be used to non-invasively measure the optical absorption and reduced scattering coefficients of tissue at depths of several millimeters. A multi-frequency DPDW system with one light source and one detector was constructed so that light is focused onto the tissue surface using an optical fiber and lens mounted to a digitally-controlled actuator which changes the distance between light source and detector. A variable RF generator enables the modulation frequency to be selected between 50 to 400MHz. The ability to digitally control both source-detector separation distance and modulation frequency allows for virtually unlimited number of data points, enabling precise selection of the volume and depth of tissue that will be characterized. Suspensions of Intralipid and india ink with known absorption and reduced scattering coefficients were used as optical phantoms to assess device accuracy. Solid silicon phantoms were formulated for stability testing. Standard deviations for amplitude and phase shift readings were found to be 0.9% and 0.2 degrees respectively, over a one hour period. The ability of the system to quantify tissue damage in vivo at multiple depths was tested in a porcine burn model.

New design of the excitation light source that can stably generate light with center wavelengths of 450nm, 530nm, 632.8nm and white light for auto-fluorescence(AF) and photodynamic diagnosis(PDD) of cancer in clinics in a single system is presented in this study. The light source consists of Xenon Lamp (300W), light guide module including motorize filter wheel equipped with optical filters with corresponding to wavelength bands, servo motor, motorize iris, a cooling system, power supply and optical transmission part for the output light. The transmission part of the light source was developed to collimate the light with desired wavelength into input of fiber optic. Output powers are obtained average 99.91 mW for 450±40 nm, 111.01 mW for 530±10nm, and 78.50 mW for 632.8±10nm.

We report a new device and algorithm that allows simultaneous monitoring of the hematocrit and plasma volume fraction of blood within the intravascular space of an optically probed volume of skin. Skin is probed with a near infrared (NIR) laser and simultaneously collecting the Rayleigh and Mie scattered light as one raw signal and the undifferentiated Raman and fluorescence emission as the second raw signal. These signals are combined using six parameters that can be obtained by either direct calculation or empirical calibration to permit monitoring of the blood in human skin (e.g. fingertips). We tested a device based on the algorithm that might be useful in allowing the early detection of blood loss for people who have no external injury but may be hemorrhaging internally. IRB allowed experiments monitoring blood in human fingertip skin in vivo during routine hemodialysis demonstrated good agreement between the experimental device and the CRIT-LINE®, an FDA approved device that is built into the dialysis machine and applies the Twersky algorithm to blood in the dialysis machine (i.e. in vitro). Based on observation of 9 different test subjects, as dialysis removes fluid from the intravascular space causing an increase in hematocrit and a decrease in plasma volume, the CRIT-LINE response is closely emulated (typical per session linear correlation r²=0.78, N=87, p<0.0001) with the new device. Calibration across subjects, the measurement of absolute hematocrit, and potential confounding factors will also be discussed.

For infants and neonates in an incubator vital signs, such as heart rate, breathing, skin temperature and blood oxygen saturation are measured by sensors and electrodes sticking to the skin. This can damage the vulnerable skin of neonates and cause infections. In addition, the wires interfere with the care and hinder the parents in holding and touching the baby. These problems initiated the search for baby friendly 'non-contact' measurement of vital signs. Using a sensitive color video camera and specially developed software, the heart rate was derived from subtle repetitive color changes. Potentially also respiration and oxygen saturation could be obtained. A thermal camera was used to monitor the temperature distribution of the whole body and detect small temperature variations around the nose revealing the respiration rate. After testing in the laboratory, seven babies were monitored (with parental consent) in the neonatal intensive care unit (NICU) simultaneously with the regular monitoring equipment. From the color video recordings accurate heart rates could be derived and the thermal images provided accurate respiration rates. To correct for the movements of the baby, tracking software could be applied. At present, the image processing was performed off-line. Using narrow band light sources also non-contact blood oxygen saturation could be measured. Non-contact monitoring of vital signs has proven to be feasible and can be developed into a real time system. Besides the application on the NICU non-contact vital function monitoring has large potential for other patient groups.

Sentinel lymph node (SLN) in vivo detection is vital in breast cancer surgery. A new near-infrared fluorescence-based surgical navigation system (SNS) imaging software, which has been developed by our research group, is presented for SLN detection surgery in this paper. The software is based on the fluorescence-based surgical navigation hardware system (SNHS) which has been developed in our lab, and is designed specifically for intraoperative imaging and postoperative data analysis. The surgical navigation imaging software consists of the following software modules, which mainly include the control module, the image grabbing module, the real-time display module, the data saving module and the image processing module. And some algorithms have been designed to achieve the performance of the software, for example, the image registration algorithm based on correlation matching. Some of the key features of the software include: setting the control parameters of the SNS; acquiring, display and storing the intraoperative imaging data in real-time automatically; analysis and processing of the saved image data. The developed software has been used to successfully detect the SLNs in 21 cases of breast cancer patients. In the near future, we plan to improve the software performance and it will be extensively used for clinical purpose.

With 332,000 operations carried out every year, the implantation of an artificial hip joint is one of the most common surgical operations performed in the US. According to prognosis which takes the demographical change into account, the number of these operations will increase in the coming years. One of the essential requirements is the perfect reconstruction of the biomechanical functions, especially the knowledge about the center of the hip rotation and the length of the leg. Based on this information it is possible to ensure the right position of the newly set leg during surgery. The aim of this work is to present and evaluate an optical measurement method in order to gather information about the center of the hip joint and the leg length. An appropriate laboratory setup has been designed and implemented in order to evaluate two different approaches: a structured light-method consisting of a DLP-Beamer or a laser source which projects defined patterns onto the patient and a marker-based system. Together with this both methods are combined with custom software to determine the hip joint center and the leg length with an accuracy of around +/- 0.2 inches. The clinical use of the tested approaches would give the surgeon the opportunity to reset the implant-parameters in the course of the surgery. In this way subsequent illnesses such as scoliotic pelvis can be prevented.

Autofluorescence characteristics of human serum albumin (HSA) are highly sensitive to its local environment. Identification and characterization of the proteins in normal and disease conditions may have great clinical implications. Aim of the present study was to understand how autofluorescence properties of HSA varies with denaturation under urea (3.0M, 6.0M, 9.0M) and guanidine hydrochloride (GnHCl) (2.0M, 4.0M, 6.0M) as well as digestion with trypsin. Towards this, we have recorded the corresponding autofluorescence spectra of HSA at 281nm laser excitation and compared the outcomes. Although, HSA contains 1 tryptophan and 17 tyrosine residues, it has shown intense autofluorescence due to tryptophan as compared to the tyrosine in native form, which may be due to the fluorescence resonance energy transfer (FRET) from tyrosine to tryptophan. As the unfolding progresses in denatured and digested forms of the protein, a clear increase in tyrosine fluorescence as compared to tryptophan was observed, which may be due to the increase of tryptophan - tyrosine separation disturbing the FRET between them resulting in differences in the overall autofluorescence properties. The decrease in tryptophan fluorescence of around 17% in urea denatured, 32% in GnHCl denatured and 96% in tryptic digested HSA was observed as compared to its native form. The obtained results show a clear decrease in FRET between tyrosine and tryptophan residues with the progression of unfolding and urea seems to be less efficient than GnHCl in unfolding of HSA. These results demonstrate the potential of autofluorescence in characterizing proteins in general and HSA in particular.

We report a new algorithm and measurement system that permits simultaneous monitoring of the hematocrit and plasma volume fraction of blood within the intravascular space of an optically probed volume of skin. The system involves probing with a near infrared laser and simultaneously collecting the Rayleigh and Mie scattered light as one raw signal and the undifferentiated Raman and fluorescence emission as the second raw signal. Those two physically independent raw signals and six parameters that can be obtained by either direct calculation or empirical calibration permit monitoring of the blood in rat paws. We tested a device based on the algorithm in the context of improving detection of blood loss for people with an early undiagnosed internal hemorrhage via real-time monitoring of signal changes with direct correlation to hematocrit. We performed experiments monitoring rat paw skin in vivo while removing blood, centrally or peripherally, and then adding replacement fluids such as Normocarb and blood. Blood removal itself elicits a predictable and consistent response, decreasing hematocrit and increasing relative plasma volume, that depends on the rate and location of removal, the total amount of blood removed, the location of monitoring, and possibly other factors as yet unknown. Similarly, replacing the blood with whole blood vs. saline consistently produces a rational range of responses. Calibration across subjects and the measurement of absolute hematocrit will also be discussed.

Accurate optical characterization of different tissue types is an important tool for potentially guiding surgeons and enabling automated robotic surgery. Multispectral imaging and analysis have been used in the literature to detect spectral variations in tissue reflectance that may be visible to the naked eye. Using this technique, hidden structures can be visualized and analyzed for effective tissue classification. Here, we investigated the feasibility of automated tissue classification using multispectral tissue analysis. Broadband reflectance spectra (200-1050 nm) were collected from nine different ex vivo porcine tissues types using an optical fiber-probe based spectrometer system. We created a mathematical model to train and distinguish different tissue types based upon analysis of the observed spectra using total principal component regression (TPCR). Compared to other reported methods, our technique is computationally inexpensive and suitable for real-time implementation. Each of the 92 spectra was cross-referenced against the nine tissue types. Preliminary results show a mean detection rate of 91.3%, with detection rates of 100% and 70.0% (inner and outer kidney), 100% and 100% (inner and outer liver), 100% (outer stomach), and 90.9%, 100%, 70.0%, 85.7% (four different inner stomach areas, respectively). We conclude that automated tissue differentiation using our multispectral tissue analysis method is feasible in multiple ex vivo tissue specimens. Although measurements were performed using ex vivo tissues, these results suggest that real-time, in vivo tissue identification during surgery may be possible.

In this study, we describe a direct fit photon-tissue interaction model to quantitatively analyze reflectance spectra of biological tissue samples. The model rapidly extracts biologically-relevant parameters associated with tissue optical scattering and absorption. This model was employed to analyze reflectance spectra acquired from freshly excised human pancreatic pre-cancerous tissues (intraductal papillary mucinous neoplasm (IPMN), a common precursor lesion to pancreatic cancer). Compared to previously reported models, the direct fit model improved fit accuracy and speed. Thus, these results suggest that such models could serve as real-time, quantitative tools to characterize biological tissues assessed with reflectance spectroscopy.