Michael S. D. Smith

Multimodal nonlinear optical imaging of atherosclerotic plaque development in myocardial infarction-prone rabbits.

Label-free imaging of bulk arterial tissue is demonstrated using a multimodal nonlinear optical microscope based on a photonic crystal fiber and a single femtosecond oscillator operating at 800 nm. Colocalized imaging of extracellular elastin fibers, fibrillar collagen, and lipid-rich structures within aortic tissue obtained from atherosclerosis-prone myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbits is demonstrated through two-photon excited fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering, respectively. These images are shown to differentiate healthy arterial wall, early atherosclerotic lesions, and advanced plaques. Clear pathological changes are observed in the extracellular matrix of the arterial wall and correlated with progression of atherosclerotic disease as represented by the age of the WHHLMI rabbits.

Differentiating atherosclerotic plaque burden in arterial tissues using femtosecond CARS-based multimodal nonlinear optical imaging

A femtosecond CARS-based nonlinear optical microscope was used to simultaneously image extracellular structural proteins and lipid-rich structures within intact aortic tissue obtained from myocardial infarction-prone Watanabe heritable hyperlipidemic rabbits (WHHLMI). Clear differences in the NLO microscopic images were observed between healthy arterial tissue and regions dominated by atherosclerotic lesions. In the current ex-vivo study, we present a single parameter based on intensity changes derived from multi-channel NLO image to classify plaque burden within the vessel. Using this parameter we were able to differentiate between healthy regions of the vessel and regions with plaque, as well as distinguish plaques relative to the age of the WHHLMI rabbit.

Evaluation of texture parameters for the quantitative description of multimodal nonlinear optical images from atherosclerotic rabbit arteries

The composition and structure of atherosclerotic lesions can be directly related to the risk they pose to the patient. Multimodal nonlinear optical (NLO) microscopy provides a powerful means to visualize the major extracellular components of the plaque that critically determine its structure. Textural features extracted from NLO images were investigated for their utility in providing quantitative descriptors of structural and compositional changes associated with plaque development. Ten texture parameters derived from the image histogram and gray level co-occurrence matrix were examined that highlight specific structural and compositional motifs that distinguish early and late stage plaques. Tonal-texture parameters could be linked to key histological features that characterize vulnerable plaque: the thickness and density of the fibrous cap, size of the atheroma, and the level of inflammation indicated through lipid deposition. Tonal and texture parameters from NLO images provide objective metrics that correspond to structural and biochemical changes that occur within the vessel wall in early and late stage atherosclerosis.

Direct global adjustment methods for endoscopic mosaicking

Endoscopy is an invaluable tool for several surgical and diagnostic applications. It permits minimally invasive visualization of internal structures thus involving little or no injury to internal structures. This method of visualization however restricts the size of the imaging device and therefore compromises on the field of view captured in a single image. The problem of a narrow field of view can be solved by capturing video sequences and stitching them to generate a mosaic of the scene under consideration. Registration of images in the sequence is therefore a crucial step. Existing methods compute frame-to-frame registration estimates and use these to resample images in order to generate a mosaic. The complexity of the appearance of internal structures and accumulation of registration error in frame to frame estimates however can be large enough to cause a cumulative drift that can misrepresent the scene. These errors can be reduced by application of global adjustment schemes. In this paper, we present a set of techniques for overcoming this problem of drift for pixel based registration in order to achieve global consistency of mosaics. The algorithm uses the frame-to-frame estimate as an initialization and subsequently corrects these estimates by setting up a large scale optimization problem which simultaneously solves for all corrections of estimates. In addition we set up a graph and introduce loop closure constraints in order to ensure consistency of registration. We present our method and results in semi global and fully global graph based adjustment methods as well as validation of our results.

Nonlinear Optical Measurements of the Artery Wall: Parameters Related to the Progression of Atherosclerosis

Nonlinear Optical Measurements of the Artery Wall: Parameters Related to the Progression of Atherosclerosis Nonlinear optical (NLO) microscopy is used to follow key structural and biochemical changes associated with the progression of atherosclerosis. Arteries from WHHL-MI rabbits are examined using a 3 channel NLO microscope that can simultaneously monitor the coherent anti-stokes Raman scattered light (CARS), the two-photon excited fluorescence (TPEF) and the second harmonic generation (SHG) from a sample. Distinct differences in the nonlinear optical signals are observed that correlate with the age of the vessel and the presence of atherosclerotic plaque. These differences are attributed to the changing extracellular matrix and the increased lipid deposition associated with plaque development. The capability of NLO to perform 3D sectioning in thick highly scattering vessels in order to visualize structural details of the artery wall and highlight vessel pathology is demonstrated. These features make NLO a potentially valuable tool to help understand the progression of atherosclerosis.

Semi-Parametric Estimation in the Compositional Modeling of Multicomponent Systems from Raman Spectroscopic Data:

Identification and quantification of molecular species are central applications of molecular spectroscopy. In complex multicomponent systems like tissue samples, linear parametric models are often used to estimate the relative concentrations of the biochemical components of the sample. In situations where not all of the components of the sample are known or modeled, such parametric models can suffer from omitted variable bias and result in skewed estimates of component concentrations. We propose a semi-parametric approach that tries to avoid this omitted variable bias by effectively including unknown covariates as a non-parametric term in the regression equation. Constituent concentrations estimated with such partial linear models should outperform strict parametric linear models when the user has limited information on the composition of a multi-constituent system.

Perception of size and shape in stereoscopic 3D imagery

This paper explores the mathematical relationships between the scene geometry, camera parameters, and viewing environment and their influence on the viewers perception of 3D. The current practice of using horizontal image translation to set convergence has an effect on the shape ratio and 3D magnification factor of the resulting images and is not well understood by the industry. This paper examines the gap between the creative processes used by stereographers and the mathematical relationships affected by those creative processes. Examples images varying the aforementioned parameters will be demonstrated.

Development of a laser speckle imaging system for measuring relative blood flow velocity

Determining the viability of damaged or surgically reconstructed tissue is critical in most plastic and reconstructive surgery procedures. Information about tissue blood flow in the region in question can make this determination much easier. Laser speckle imaging (LSI) is one technique that could potentially aid in making this determination. LSI is a non-contact full-field imaging technique with simultaneous high spatial and temporal resolution. Tissue is illuminated with diffuse red laser light and the spatial and/or temporal statistics of the resulting speckle pattern can be used to calculate relative flow velocities. We have developed a LSI system that produces relative velocity blood flow images. Bench tests of the system indicate that it may be used to distinguish between normal, decreased, and increased blood flow states of a human finger. The system has also been used to take some initial laboratory measurements using an animal model - an epigastric free flap on a rat. Preliminary results indicate that the method may be used to distinguish states of venous or arterial occlusion from unoccluded states of the skin flap. While further experimentation is necessary, these initial results indicate that LSI could be a useful aid to the plastic surgeon for assessing tissue viability.

Quantitative nonlinear optical assessment of atherosclerosis progression in rabbits.

Quantification of atherosclerosis has been a challenging task owing to its complex pathology. In this study, we validated a quantitative approach for assessing atherosclerosis progression in a rabbit model using a numerical matrix, optical index for plaque burden, derived directly from the nonlinear optical microscopic images captured on the atherosclerosis-affected blood vessel. A positive correlation between this optical index and the severity of atherosclerotic lesions, represented by the age of the rabbits, was established based on data collected from 21 myocardial infarction-prone Watanabe heritable hyperlipidemic rabbits with age ranging between new-born and 27 months old. The same optical index also accurately identified high-risk locations for atherosclerotic plaque formation along the entire aorta, which was validated by immunohistochemical fluorescence imaging.

Using multimodal femtosecond CARS imaging to determine plaque burden in luminal atherosclerosis

Luminal atherosclerosis imaging was demonstrated by multimodal femtosecond CARS microscopy (MM-CARS). Using a myocardial infarction-prone rabbit model of atherosclerosis, this study demonstrated the utility of multimodal CARS imaging in determining atherosclerotic plaque burden through two types of image analysis procedures. Firstly, multimodal CARS images were evaluated using a signal-intensity parameter based on intensity changes derived from the multi-channel data (e.g. TPEF, SHG and CARS) to classify plaque burden within the vessel. Secondly, the SHG images that mainly correspond to collagen fibrils were evaluated using a texture analysis model based on the first-order statistical (FOS) parameters of the image histogram. Correlation between FOS parameters of collagen images with atherosclerosis plaque burden was established. A preliminary study of using spectroscopic CARS in identifying the different lipid components within the plaque was also discussed.