Bio-Inspired Multi-Spectral and Polarization Imaging Sensors for Image-Guided Surgery

Image-guided surgery (IGS) can enhance cancer treatment by decreasing, and ideally eliminating, positive tumor margins and iatrogenic damage to healthy tissue. Current state-of-the-art near-infrared fluorescence imaging systems are bulky, costly, lack sensitivity under surgical illumination, and lack co-registration accuracy between multimodal images. As a result, an overwhelming majority of physicians still rely on their unaided eyes and palpation as the primary sensing modalities to distinguish cancerous from healthy tissue. In my thesis, I have addressed these challenges in IGC by mimicking the visual systems of several animals to construct low power, compact and highly sensitive multi-spectral and color-polarization sensors. I have realized single-chip multi-spectral imagers with 1000-fold higher sensitivity and 7-fold better spatial co-registration accuracy compared to clinical imaging systems in current use by monolithically integrating spectral tapetal and polarization filters with an array of vertically stacked photodetectors. These imaging sensors yield the unique capabilities of imaging simultaneously color, polarization, and multiple fluorophores for near-infrared fluorescence imaging. Preclinical and clinical data demonstrate seamless integration of this technologies in the surgical work flow while providing surgeons with real-time information on the location of cancerous tissue and sentinel lymph nodes, respectively. Due to its low cost, the bio-inspired sensors will provide resource-limited hospitals with much-needed technology to enable more accurate value-based health care

Using Fluorescence – Polarization Endoscopy in Detection of Precancerous and Cancerous Lesions in Colon and Pancreatic Cancer

Colitis-associated cancer (CAC) arises from premalignant flat lesions of the colon, which are difficult to detect with current endoscopic screening approaches. We have developed a complementary fluorescence and polarization reporting strategy that combines the unique biochemical and physical properties of dysplasia and cancer for real time detection of these lesions. Utilizing a new thermoresponsive sol-gel formulation with targeted molecular probe allowed topical application and detection of precancerous and cancerous lesions during endoscopy. Incorporation of nanowire-filtered polarization imaging into NIR fluorescence endoscopy served as a validation strategy prior to obtaining biopsies. In order to reduce repeat surgeries arising from incomplete tumor resection, we demonstrated the efficacy of the targeted molecular probe towards margins of sporadic colorectal cancer (SCC). Fluorescence-polarization microscopy using circular polarized (CP) light served as a rapid, supplementary tool for assessment and validation of excised tissue to ensure complete tumor resection for examining tumor margins prior to H&E-based pathological diagnosis. We extended our platform towards non-invasive directed detection of pancreatic cancer utilizing fluorescence molecular tomography (FMT) and NIR laparoscopy using identified targeted molecular probe. We were able to non-invasively distinguished between pancreatitis and pancreatic cancer and guide pancreatic tumor resection using NIR laparoscopy

Endoscopic Optical Coherence Tomography: Design and Application

This thesis presents an investigation on endoscopic optical coherence tomography (OCT). As a noninvasive imaging modality, OCT emerges as an increasingly important diagnostic tool for many clinical applications. Despite of many of its merits, such as high resolution and depth resolvability, a major limitation is the relatively shallow penetration depth in tissue (about 2∼3 mm). This is mainly due to tissue scattering and absorption. To overcome this limitation, people have been developing many different endoscopic OCT systems. By utilizing a minimally invasive endoscope, the OCT probing beam can be brought to the close vicinity of the tissue of interest and bypass the scattering of intervening tissues so that it can collect the reflected light signal from desired depth and provide a clear image representing the physiological structure of the region, which can not be disclosed by traditional OCT. In this thesis, three endoscope designs have been studied. While they rely on vastly different principles, they all converge to solve this long-standing problem. A hand-held endoscope with manual scanning is first explored. When a user is holding a hand- held endoscope to examine samples, the movement of the device provides a natural scanning. We proposed and implemented an optical tracking system to estimate and record the trajectory of the device. By registering the OCT axial scan with the spatial information obtained from the tracking system, one can use this system to simply ‘paint’ a desired volume and get any arbitrary scanning pattern by manually waving the endoscope over the region of interest. The accuracy of the tracking system was measured to be about 10 microns, which is comparable to the lateral resolution of most OCT system. Targeted phantom sample and biological samples were manually scanned and the reconstructed images verified the method. Next, we investigated a mechanical way to steer the beam in an OCT endoscope, which is termed as Paired-angle-rotation scanning (PARS). This concept was proposed by my colleague and we further developed this technology by enhancing the longevity of the device, reducing the diameter of the probe, and shrinking down the form factor of the hand-piece. Several families of probes have been designed and fabricated with various optical performances. They have been applied to different applications, including the collector channel examination for glaucoma stent implantation, and vitreous remnant detection during live animal vitrectomy. Lastly a novel non-moving scanning method has been devised. This approach is based on the EO effect of a KTN crystal. With Ohmic contact of the electrodes, the KTN crystal can exhibit a special mode of EO effect, termed as space-charge-controlled electro-optic effect, where the carrier electron will be injected into the material via the Ohmic contact. By applying a high voltage across the material, a linear phase profile can be built under this mode, which in turn deflects the light beam passing through. We constructed a relay telescope to adapt the KTN deflector into a bench top OCT scanning system. One of major technical challenges for this system is the strong chromatic dispersion of KTN crystal within the wavelength band of OCT system. We investigated its impact on the acquired OCT images and proposed a new approach to estimate and compensate the actual dispersion. Comparing with traditional methods, the new method is more computational efficient and accurate. Some biological samples were scanned by this KTN based system. The acquired images justified the feasibility of the usage of this system into a endoscopy setting. My research above all aims to provide solutions to implement an OCT endoscope. As technology evolves from manual, to mechanical, and to electrical approaches, different solutions are presented. Since all have their own advantages and disadvantages, one has to determine the actual requirements and select the best fit for a specific application.</p

Dynamic Hyperspectral and Polarized Endoscopic Imaging

The health of rich, developed nations has seen drastic improvement in the last two centuries. For it to continue improving at a similar rate new or improved diagnostic and treatment technologies are required, especially for those diseases such as cancer which are forecast to constitute the majority of disease burden in the future. Optical techniques such as microscopy have long played their part in the diagnostic process. However there are several new biophotonic modalities that aim to exploit various interactions between light and tissue to provide enhanced diagnostic information. Many of these show promise in a laboratory setting but few have progressed to a clinical setting. We have designed and constructed a flexible, multi-modal, multi-spectral laparoscopic imaging system that could be used to demonstrate several different techniques in a clinical setting. The core of this system is a dynamic hyperspectral illumination system based around a supercontinuum laser and Digital Micromirror Device that can provide specified excitation light in the visible and near infra-red ranges. This is a powerful tool for spectroscopic techniques as it is not limited to interrogating a fixed range of wavelengths and can switch between excitation bands instantaneously. The excitation spectra can be customised to match particular fluorophores or absorption features, introducing new possibilities for spectral imaging. A standard 10 mm diameter rigid endoscope was incorporated into the system to reduce cost and demonstrate compatibility with existing equipment. The polarization properties of two commercial endoscopes were characterised and found to be unsuited to current polarization imaging techniques as birefringent materials used in their construction introduce complex, spatially dependent transformations of the polarization state. Preliminary exemplar data from phantoms and ex vivo tissue was collected and the feasibility and accuracy of different analysis techniques demonstrated including multiple class classification algorithms. Finally, a novel visualisation method was implemented in order to display the complex hyperspectral data sets in a meaningful and intuitive way to the user

Vision Sensors and Edge Detection

Vision Sensors and Edge Detection book reflects a selection of recent developments within the area of vision sensors and edge detection. There are two sections in this book. The first section presents vision sensors with applications to panoramic vision sensors, wireless vision sensors, and automated vision sensor inspection, and the second one shows image processing techniques, such as, image measurements, image transformations, filtering, and parallel computing

Wavelength swept spectrally encoded confocal microscopy for biological and clinical applications

Thesis (Ph. D.)--Harvard-MIT Division of Health Sciences and Technology, 2007.Includes bibliographical references (p. 157-168).Spectrally encoded confocal microscopy (SECM) is a technique that facilitates the incorporation of confocal microscopy into small, portable clinical instruments. This would allow in vivo evaluation of cellular and sub-cellular features in a non-destructive, minimally invasive manner. Prior studies have demonstrated the potential of the techniques as well as highlighted the need for faster acquisition rates and higher sensitivity. In this thesis, new laser sources, optical fiber arrangements and probe designs are explored to ultimately evaluate SECM's relevance as a clinical tool. Clinical imaging at cellular scales requires imaging rates on the order of tens of frames per second to reduce motion artifacts from unavoidable patient movements. Rapid SECM imaging was achieved through the development of a novel wavelength swept laser which simultaneously provided high output power (> 10mrW), narrow linewidth (10GHz), broad wavelength tuning (80 nm centered at 1310 nm) and fast repetition rates (up to 16,000 Hz), while being compact and environmentally stable. Imaging with a wavelength swept SECM system was characterized by coupling the laser to a tabletop imaging arm comprising a high density holographic grating, a galvanometer mounted mirror and a 0.9 NA water immersion microscope objective.(cont.) Rapid SECM imaging is performed at a transverse resolution of 1.4 microns, axial resolution of 6 microns over a field of view of 440x440 microns and allows subcellular imaging ex vivo (excised specimens) and in vivo (human skin). A study on 40 excised head and neck specimens showed that SECM has the potential to perform tissue identification, but also revealed the presence of speckle noise due to the coherent nature of the illumination and collection schemes through a single mode optical fiber. A partially coherent system based on single mode fiber for illumination and multimode fiber for detection was simulated, implemented and tested to find adequate balance between attenuation of speckle noise and conservation of resolution. A coupling of 20 modes was found to reduce speckle by a factor 4.5 with a minimal sectioning penalty of 0.25, while allowing a signal increase of 8dB. This improvement in sensitivity allowed SECM table top system to be used for investigations in developmental biology where Dual clad fibers (DCF) were previously shown to allow partially coherent endoscopic imaging, using the single mode core for illumination and inner clad for multimodal collection.(cont.) Commercially available DCF's which propagate thousands of modes are ill suited for confocal endoscopes as collecting such a number of modes would destroy the axial resolution. Based on results from the previous section and through modal analysis, a DCF was designed, drawn - via a collaboration with Boston University Photonics Center -, and tested for use with SECM. The prototype DCF yielded promising results (3 fold speckle attenuation, optical sectioning degradation of 0.85), and showed the need for implementation of better coupling mechanisms to take advantage of increased signal collection. Finally, a portable SECM system was built for in vivo evaluation of pediatric vocal fold. A preliminary study on porcine and cadaveric tissue showed that SECM can distinguish between epithelium, superior and intermediate layers of the lamina propria, which could help elucidate the development mechanism of the voice apparatus if performed in vivo. The handheld instrument comprises a custom grating scanner imaging the scanning pivot onto the back pupil of a high NA microscope objective. The imaging tube can easily be interchanged to accommodate geometrical constraints imposed by different age groups.(cont.) The probe, currently under review by the biomedical engineering committee, revealed cellular and sub cellular details of human skin in vivo at depth and acquisition rates sufficient to capture blood cells flowing through capillaries. Through major improvements in acquisition speeds, sensitivity, and speckle appearance, this work established SECM as a potent clinical and biological imaging tool. Ultimate confirmation will be revealed through in vivo studies to come, but limitations are likely to be of engineering nature rather than from physical considerations. Future work should explore the possibility to combine SECM with other contrast mechanisms to provide imaging with increased specificity.by Caroline Boudoux.Ph.D

Development of a miniaturized microscope for depth-scanning imaging at subcellular resolution in freely behaving animals

Le fonctionnement du cerveau humain est fascinant. En seulement quelques millisecondes, des milliards de neurones synchronisés perçoivent, traitent et redirigent les informations permettant le contrôle de notre corps, de nos sentiments et de nos pensées. Malheureusement, notre compréhension du cerveau reste limitée et de multiples questions physiologiques demeurent. Comment sont exactement reliés le fonctionnement neuronal et le comportement humain ? L’imagerie de l’activité neuronale au moyen de systèmes miniatures est l’une des voies les plus prometteuses permettant d’étudier le cerveau des animaux se déplaçant librement. Cependant, le développement de ces outils n’est pas évident et de multiples compromis techniques doivent être faits pour arriver à des systèmes suffisamment petits et légers. Les outils actuels ont donc souvent des limitations concernant leurs caractéristiques physiques et optiques. L’un des problèmes majeur est le manque d’une lentille miniature électriquement réglable et à faible consommation d’énergie permettant l’imagerie avec un balayage en profondeur. Dans cette thèse, nous proposons un nouveau type de dispositif d’imagerie miniature qui présente de multiples avantages mécaniques, électriques et optiques par rapport aux systèmes existants. Le faible poids, la petite dimension, la capacité de moduler électriquement la distance focale à l’aide d’une lentille à cristaux liquides (CL) et la capacité d’imager des structures fines sont au cœur des innovations proposées. Dans un premier temps, nous présenterons nos travaux (théoriques et expérimentaux) de conception, assemblage et optimisation de la lentille à CL accordable (TLCL, pour tunable liquid crystal lens). Deuxièmement, nous présenterons la preuve de concept macroscopique du couplage optique entre la TLCL et la lentille à gradient d’indice (GRIN, pour gradient index) en forme d’une tige. Utilisant le même système, nous démontrerons la capacité de balayage en profondeur dans le cerveau des animaux anesthésiés. Troisièmement, nous montrerons un dispositif d’imagerie (2D) miniature avec de nouvelles caractéristiques mécaniques et optiques permettant d’imager de fines structures neuronales dans des tranches de tissus cérébraux fixes. Enfin, nous présenterons le dispositif miniaturisé, avec une TLCL intégrée. Grâce à notre système, nous obtenons ≈ 100 µm d’ajustement électrique de la profondeur d’imagerie qui permet d’enregistrer l’activité de fines structures neuronales lors des différents comportements (toilettage, marche, etc.) de la souris.The functioning of the human brain is fascinating. In only a few milliseconds, billions of finely tuned and synchronized neurons perceive, process and exit the information that drives our body, our feelings and our thoughts. Unfortunately, our understating of the brain is limited and multiple physiological questions remain. How exactly are related neural functioning and human behavior ? The imaging of the neuronal activity by means of miniaturized systems is one of the most promising avenues allowing to study the brain of the freely moving subjects. However, the development of these tools is not obvious and multiple technical trade-offs must be made to build a system that is sufficiently small and light. Therefore, the available tools have different limitations regarding their physical and optical characteristics. One of the major problems is the lack of an electrically adjustable and energy-efficient miniature lens allowing to scan in depth. In this thesis, we propose a new type of miniature imaging device that has multiple mechanical, electrical and optical advantages over existing systems. The low weight, the small size, the ability to electrically modulate the focal distance using a liquid crystal (LC) lens and the ability to image fine structures are among the proposed innovations. First, we present our work (theoretical and experimental) of design, assembling and optimization of the tunable LC lens (TLCL). Second, we present the macroscopic proof-of-concept optical coupling between the TLCL and the gradient index lens (GRIN) in the form of a rod. Using the same system, we demonstrate the depth scanning ability in the brain of anaesthetized animals. Third, we show a miniature (2D) imaging device with new mechanical and optical features allowing to image fine neural structures in fixed brain tissue slices. Finally, we present a state-of-the-art miniaturized device with an integrated TLCL. Using our system, we obtain a ≈ 100 µm electrical depth adjustment that allows to record the activity of fine neuronal structures during the various behaviours (grooming, walking, etc.) of the mouse

TOWARDS HIGH RESOLUTION ENDOSCOPIC OPTICAL COHERENCE TOMOGRAPHY FOR IMAGING INTERNAL ORGANS

Optical coherence tomography (OCT) is a light based interferometric imaging technique that can provide high resolution (5-20 µm at 1300 nm), depth resolved, images in real-time. With recent advances in portable low coherent light sources for OCT it is now possible to achieve ultrahigh axial resolutions (≤ 3 µm) by moving to shorter central wavelengths such as 800 nm while utilizing a broad spectral bandwidth. Our goal was to push ultrahigh resolution OCT technology to in vivo imaging of internal organs for endoscopic assessment of tissue microstructure. This dissertation is separated into technological developments and biomedical imaging studies. Technological developments in this dissertation included development of a high speed, ultrahigh resolution distal scanning catheter. This catheter was based upon a miniature DC micromotor capable of rotational velocities in excess of 100 rps, a diffractive compound lens design that minimized chromatic aberrations, and a mechanical assembly that limited field of view blockage to less than 7.5% and maintained an outer diameter of 1.78 mm (with plastic sheath). In conjunction with the algorithm described in chapter 4 to correct for non-uniform rotational distortion, the overall imaging system was capable of high quality endoscopic imaging of internal organs in vivo. Equipped with the ultrahigh resolution endoscopic OCT system, imaging was performed in small airways and colorectal cancer. Imaging results demonstrated the ability to directly visualization of microstructural details such as airway smooth muscle in the small airways representing a major step forward in pulmonary imaging. With the ability to visualize airway smooth muscle, morphological changes in COPD and related diseases can be further investigated. Additionally, longitudinal changes in an ETBF induced colon cancer model in APCMin mice were studied as well. Quantitative assessment of tissue microarchitecture was performed by measuring the attenuation coefficient to find a bimodal distribution separating normal healthy tissue from polyps. Finally, results from two additional projects were also demonstrated. Chapter 7 shows some results from vascular imaging in a tumor angiogenesis model and middle cerebral artery occlusion model. Chapter 8 describes an endoscopic multimodal OCT and fluorescence imaging platform with results in ex vivo rabbit esophagus

Computational Multispectral Endoscopy

Minimal Access Surgery (MAS) is increasingly regarded as the de-facto approach in interventional medicine for conducting many procedures this is due to the reduced patient trauma and consequently reduced recovery times, complications and costs. However, there are many challenges in MAS that come as a result of viewing the surgical site through an endoscope and interacting with tissue remotely via tools, such as lack of haptic feedback; limited field of view; and variation in imaging hardware. As such, it is important best utilise the imaging data available to provide a clinician with rich data corresponding to the surgical site. Measuring tissue haemoglobin concentrations can give vital information, such as perfusion assessment after transplantation; visualisation of the health of blood supply to organ; and to detect ischaemia. In the area of transplant and bypass procedures measurements of the tissue tissue perfusion/total haemoglobin (THb) and oxygen saturation (SO2) are used as indicators of organ viability, these measurements are often acquired at multiple discrete points across the tissue using with a specialist probe. To acquire measurements across the whole surface of an organ one can use a specialist camera to perform multispectral imaging (MSI), which optically acquires sequential spectrally band limited images of the same scene. This data can be processed to provide maps of the THb and SO2 variation across the tissue surface which could be useful for intra operative evaluation. When capturing MSI data, a trade off often has to be made between spectral sensitivity and capture speed. The work in thesis first explores post processing blurry MSI data from long exposure imaging devices. It is of interest to be able to use these MSI data because the large number of spectral bands that can be captured, the long capture times, however, limit the potential real time uses for clinicians. Recognising the importance to clinicians of real-time data, the main body of this thesis develops methods around estimating oxy- and deoxy-haemoglobin concentrations in tissue using only monocular and stereo RGB imaging data

Seeing the Big Picture: System Architecture Trends in Endoscopy and LED-Based hyperspectral Subsystem Intergration

Early-stage colorectal lesions remain difficult to detect. Early development of neoplasia tends to be small (less than 10 mm) and flat and difficult to distinguish from surrounding mucosa. Additionally, optical diagnosis of neoplasia as benign or malignant is problematic. Low rates of detection of these lesions allow for continued growth in the colorectum and increased risk of cancer formation. Therefore, it is crucial to detect neoplasia and other non-neoplastic lesions to determine risk and guide future treatment. Technology for detection needs to enhance contrast of subtle tissue differences in the colorectum and track multiple biomarkers simultaneously. This work implements one such technology with the potential to achieve the desired multi-contrast outcome for endoscopic screenings: hyperspectral imaging. Traditional endoscopic imaging uses a white light source and a RGB detector to visualize the colorectum using reflected light. Hyperspectral imaging (HSI) acquires an image over a range of individual wavelength bands to create an image hypercube with a wavelength dimension much deeper and more sensitive than that of an RGB image. A hypercube can consist of reflectance or fluorescence (or both) spectra depending on the filtering optics involved. Prior studies using HSI in endoscopy have normally involved ex vivo tissues or xiv optics that created a trade-off between spatial resolution, spectral discrimination and temporal sampling. This dissertation describes the systems design of an alternative HSI endoscopic imaging technology that can provide high spatial resolution, high spectral distinction and video-rate acquisition in vivo. The hyperspectral endoscopic system consists of a novel spectral illumination source for image acquisition dependent on the fluorescence excitation (instead of emission). Therefore, this work represents a novel contribution to the field of endoscopy in combining excitation-scanning hyperspectral imaging and endoscopy. This dissertation describes: 1) systems architecture of the endoscopic system in review of previous iterations and theoretical next-generation options, 2) feasibility testing of a LED-based hyperspectral endoscope system and 3) another LED-based spectral illuminator on a microscope platform to test multi-spectral contrast imaging. The results of the architecture point towards an endoscopic system with more complex imaging and increased computational capabilities. The hyperspectral endoscope platform proved feasibility of a LED-based spectral light source with a multi-furcated solid light guide. Another LED-based design was tested successfully on a microscope platform with a dual mirror array similar to telescope designs. Both feasibility tests emphasized optimization of coupling optics and combining multiple diffuse light sources to a common output. These results should lead to enhanced imagery for endoscopic tissue discrimination and future optical diagnosis for routine colonoscopy