More and more researchers and clinicians are looking to molecular sensing to predict how cells will behave, seeking the answers to questions like "will these tumor cells become malignant?" or "how will these cells respond to chemotherapy?" Optical methods are attractive for answering these questions because optical radiation is safer and less expensive than alternative methods, such as CT which uses X-ray radiation, PET/SPECT which use gamma radiation, or MRI which is expensive and only available in a hospital setting. In this dissertation, three distinct optical methods are explored to detect at the molecular level: optical coherence tomography (OCT), laser-induced fluorescence (LIF), and optical polarimetry. OCT has the capability to simultaneously capture anatomical information as well as molecular information using targeted contrast agents such as gold nanoshells. LIF is less useful for capturing anatomical information, but it can achieve significantly better molecular sensitivity with the use of targeted fluorescent dyes. Optical polarimetry has potential to detect the concentration of helical molecules, such as glucose. All of these methods are noninvasive or minimally invasive.The work is organized into four specific aims. The first is the design and implementation of a fast, high resolution, endoscopic OCT system to facilitate minimally invasive mouse colon imaging. The second aim is to demonstrate the utility of this system for automatically identifying tumor lesions based on tissue microstructure. The third is to demonstrate the use of contrast agents to detect molecular expression using OCT and LIF. The last aim is to demonstrate a new method based on optical polarimetry for noninvasive glucose sensing.

Measuring the Mueller matrix with optical coherence tomography (OCT) makes it possible to acquire the complete polarization properties of scattering media with three-dimensional spatial resolution. We first proved that the measured degree-of-polarization (DOP) of the backscattered light by OCT remains unity-a conclusion that validated the use of Jones calculus in OCT. A multi-channel Mueller-matrix OCT system was then built to measure the Jones-matrix, which can be transformed into a Mueller matrix, images of scattering biological tissues accurately with single depth scan. We showed that when diattenuation is negligible, the round-trip Jones matrix represents a linear retarder, which is the foundation of conventional PS-OCT, and can be calculated with a single incident polarization state although the one-way Jones matrix generally represents an elliptical retarder; otherwise, two incident polarization states are needed. We discovered the transpose symmetry in the roundtrip Jones matrix, which is critical for eliminating the arbitrary phase difference between the two measured Jones vectors corresponding to the two incident polarization states to yield the correct Jones matrix. We investigated the various contrast mechanisms provided by Mueller-matrix OCT. Our OCT system for the first time offers simultaneously comprehensive polarization contrast mechanisms including the amplitude of birefringence, the orientation of birefringence, and the diattenuation in addition to the polarization-independent intensity contrast, all of which can be extracted from the measured Jones or the equivalent Mueller matrix. The experimental results obtained from rat skin samples, show that Mueller OCT provides complementary structural and functional information on biological samples and reveal that polarization contrast is more sensitive to thermal degeneration of biological tissues than amplitude-based contrast. Finally, an optical-fiber-based multi-channel Mueller-matrix OCT was built and a new rigorous algorithm was developed to retrieve the calibrated polarization properties of a sample. For the first time to our knowledge, fiber-based polarization-sensitive OCT was dynamically calibrated to eliminate the polarization distortion caused by the single-mode optical fiber in the sample arm, thereby overcoming a key technical impediment to the application of optical fibers in this technology.

The objective of this study is to apply digital image processing (DIP) techniques to optical coherence tomography (OCT) images and develop computer-based non-subjective quantitative analysis, which can be used as diagnostic aids in early detection of dental caries. This study first compares speckle reduction effects on raw OCT image data by implementing spatial-domain and transform-domain speckle filtering. Then region-based contour search and global thresholding techniques examine digital OCT images with possible lesions to identify and highlight the presence of features indicating early stage dental caries. The outputs of these processes, which explore the combination of image restoration and segmentation, can be used to distinguish lesion from normal tissue and determine the characteristics prior to, during, and following treatments. The combination of image processing and analysis techniques in this thesis shows potential of detecting early stage caries lesion successfully.

Obtaining higher depth of imaging is an important goal in Optical Coherence Tomography (OCT) systems. One of the main factors that affect the depth of OCT imaging is the presence of noise. That’s why the study of noise statistics is an important problem. In the first part of this thesis we obtain an empirical estimate of the second order statistics of noise by using a sequence of Time domain (TD) OCT images. These estimates confirm the non-stationary nature of noise in TD-OCT. In the second part of the thesis these estimates are used to design a depth-dependent matched filter to maximize the Signal-to-Noise Ratio (SNR) and increase the Contrast-to-Noise Ratio (CNR) in TD-OCT. By applying our filter to TD-OCT images of both vascular rabbit tissue and a human tooth, both SNR and CNR were increased and a higher imaging depth was achieved.

About 50% of cancer patients are treated with X-ray radiation therapy; however, with current treatment feedback, the effects and the efficacy of the treatment are generally detected several weeks/months after treatment completion. This makes the adjustment of the treatment based on early response, and identification of non-responding patients, nearly impossible. In this thesis a novel method combining optical coherence tomography and a gamut of image analysis methods is explored as a potential approach to detecting tissue variability. Applying texture analysis to the optical coherence tomography images may allow for the tracking of radiation therapy induced cell microstructural changes in cancer patients and help in the adjustment of treatment based on early response.

About 50% of cancer patients are treated with X-ray radiation therapy; however, with current treatment feedback, the effects and the efficacy of the treatment are generally detected several weeks/months after treatment completion. This makes the adjustment of the treatment based on early response, and identification of non-responding patients, nearly impossible. In this thesis a novel method combining optical coherence tomography and a gamut of image analysis methods is explored as a potential approach to detecting tissue variability. Applying texture analysis to the optical coherence tomography images may allow for the tracking of radiation therapy induced cell microstructural changes in cancer patients and help in the adjustment of treatment based on early response.

Optical imaging has been developed quickly in the past decades because it has become an important research tool in biology, biochemistry, and biomedical sciences. Coherence domain optical imaging is one of the well developed optical imaging modalities, as it provides high resolution and long penetration depth. In this dissertation, we will report our work on development of molecular contrast in coherence domain optical imaging. In order to image important molecules which are poor fluorophores, we developed a high resolution molecular imaging technique, pump-probe optical coherence microscopy (PPOCM), which does not rely on fluorescent tags. PPOCM is the fusion of Pump-Probe spectroscopy and optical coherence microscopy (OCM). We have demonstrated the prototype system on a fixed human skin sample containing a nodular melanoma. The results indicate that PPOCM can clearly provide strong contrast between the melanotic and amelanotic regions. This technique can be applied to early diagnosis of melanoma and the mapping of tumor margins during excision. It also can be extended to any biological chromophore with a known absorption spectrum and sufficient concentration. In order to differentiate further multiple chromophores, we developed a spectrally resolved two color pump-probe Optical Coherence Microscopy (SRPPOCM). We showed the prototype system on a red hair and a black hair. Our preliminary results show that the SRPPOCM technique could provide a contrast between pheomelanin and eumelanin. This technique could be used potentially as a clinical tool for diagnosing different progression stages of melanoma. This technique could also be applied to differentiate other mixed chromophores. Second harmonic optical coherence tomography (SHOCT) is non-linear high resolution optical molecular imaging modality which is widely used in non-centrosymmetric material. However, depth ambiguity is associated with SHOCT in tissue sample because forward generated second harmonic signal does not correctly report where the second harmonic signal is generated. We studied the feasibility of collecting the backward generated second harmonic signal from nanocrystals through a Second Harmonic Optical Coherence Tomography in Fourier domain. The preliminary result shows that we can collect backward generated second harmonic signal from nanocrystals which indicates that this technique could suppress the depth ambiguity.

<p>Clinical imaging within ophthalmology has had transformative effects on ocular health over the last century. Imaging has guided clinicians in their pharmaceutical and surgical treatments of macular degeneration, glaucoma, cataracts and numerous other pathologies. Many of the imaging techniques currently used are photography based and are limited to imaging the surface of ocular structures. This limitation forces clinicians to make assumptions about the underlying tissue which may reduce the efficacy of their diagnoses. </p><p>Optical coherence tomography (OCT) is a non-invasive, non-ionizing imaging modality that has been widely adopted within the field of ophthalmology in the last 15 years. As an optical imaging technique, OCT utilizes low-coherence interferometry to produce micron-scale three-dimensional datasets of a tissue's structure. Much of the human body consists of tissues that significantly scatter and attenuate optical signals limiting the imaging depth of OCT in those tissues to only 1-2mm. However, the ocular anterior segment is unique among human tissue in that it is primarily transparent or translucent. This allows for relatively deep imaging of tissue structure with OCT and is no longer limited by the optical scattering properties of the tissue. </p><p>This goal of this work is to develop methods utilizing OCT that offer the potential to reduce the assumptions made by clinicians in their evaluations of their patients' ocular anterior segments. We achieved this by first developing a method to reduce the effects of patient motion during OCT volume acquisitions allowing for accurate, three dimensional measurements of corneal shape. Having accurate corneal shape measurements then allowed us to determine corneal spherical and astigmatic refractive contribution in a given individual. This was then validated in a clinical study that showed OCT better measured refractive change due to surgery than other clinical devices. Additionally, a method was developed to combine the clinical evaluation of the iridocorneal angle through gonioscopy with OCT.</p> / Dissertation

Optical coherence tomography (OCT) imaging is used in an increasing number of biomedical and industrial applications. A massively parallel simulator of OCT of inhomogeneous turbid media, e.g., biological tissue, could be used as a practical tool to expedite and expand the study of the physical phenomena involving such imaging technique, as well as, to design OCT systems with enhanced performance. Our work presents the open-source implementation of this massively parallel simulator of OCT to satisfy the ever-increasing need for prompt computation of OCT signals with accuracy and flexibility. Our Monte Carlo-based simulator uses graphic processing units (GPUs) to accelerate the intensive computation of processing tens of millions of photon packets undergoing a random walk through a sample. It provides computation of both Class I diffusive reflectance due to ballistic and quasi-ballistic scattered photons and Class II diffusive reflectance due to multiple scattered photons. Our implementation was tested by comparing results with previously validated OCT simulators in multilayered and inhomogeneous (arbitrary spatial distributions) turbid media configurations. It models the objects as a tetrahedron-based mesh and implements and advanced importance sampling technique. Our massively parallel simulator of OCT speeds up the simulation of OCT signals by a factor of 40 times when compared to it central processing unit (CPU)-based sequential implementation.

Optical Coherence Tomography (OCT) is a subsurface imaging technique with many biomedical and industrial applications. In this thesis, we describe our design and implementation of a time domain OCT system. We used this system to obtain OCT images of objects that are important in different applications. We also used an existing quadrature OCT system to obtain both real and imaginary parts of an OCT image. We introduced a new interpretation of OCT images as the 2nd derivative of the scattering potential of an object. To obtain this scattering potential from its 2nd derivative, we implemented a method of definite integration in the spectral-domain. The obtained scattering potential was used to separate the scattering profile from the absorption profile of an object. We applied this new spectroscopic imaging method to quadrature OCT images of different objects.