Chao-Wei Chen

Co-registered optical coherence tomography and fluorescence molecular imaging for simultaneous morphological and molecular imaging.

We have developed a co-registered optical coherence tomography (OCT) and laser scanning fluorescence molecular imaging system. This system enables simultaneous imaging of tissue morphology and molecular information at high resolution over a several-millimeter field-of-view.

Automated quantification of microstructural dimensions of the human kidney using optical coherence tomography (OCT)

Optical coherence tomography (OCT) is a rapidly emerging imaging modality that can non-invasively provide cross-sectional, high-resolution images of tissue morphology in situ and in real-time. We previously demonstrated that OCT is capable of visualizing characteristic kidney anatomic structures, including blood vessels, uriniferous tubules, glomeruli, and renal capsules on a Munich-Wistar rat model. Because the viability of a donor kidney is closely correlated with its tubular morphology, and a large amount of image datasets are expected when using OCT to scan the entire kidney to provide a global assessment of its viability, it is necessary to develop automatic image analysis methods to quantify the spatially-resolved morphometric parameters such as tubular diameter to provide potential diagnostic information. In this study, we imaged the human kidney in vitro and quantified the diameters of hollow structures such as blood vessels and uriniferous tubules automatically. The microstructures were first segmented from cross-sectional OCT images. Then the spatially-isolated region-of-interest (ROI) was automatically selected to quantify its dimension. This method enables the automatic selection and quantification of spatially-resolved morphometric parameters. The quantification accuracy was validated, and measured features are in agreement with known kidney morphology. This work can enable studies to determine the clinical utility of OCT for kidney imaging, as well as studies to evaluate kidney morphology as a biomarker for assessing kidneys viability prior to transplantation.

Mesoscopic Fluorescence Molecular Tomography for Evaluating Engineered Tissues

Optimization of regenerative medicine strategies includes the design of biomaterials, development of cell-seeding methods, and control of cell-biomaterial interactions within the engineered tissues. Among these steps, one paramount challenge is to non-destructively image the engineered tissues in their entirety to assess structure, function, and molecular expression. It is especially important to be able to enable cell phenotyping and monitor the distribution and migration of cells throughout the bulk scaffold. Advanced fluorescence microscopic techniques are commonly employed to perform such tasks; however, they are limited to superficial examination of tissue constructs. Therefore, the field of tissue engineering and regenerative medicine would greatly benefit from the development of molecular imaging techniques which are capable of non-destructive imaging of three-dimensional cellular distribution and maturation within a tissue-engineered scaffold beyond the limited depth of current microscopic techniques. In this review, we focus on an emerging depth-resolved optical mesoscopic imaging technique, termed laminar optical tomography (LOT) or mesoscopic fluorescence molecular tomography (MFMT), which enables longitudinal imaging of cellular distribution in thick tissue engineering constructs at depths of a few millimeters and with relatively high resolution. The physical principle, image formation, and instrumentation of LOT/MFMT systems are introduced. Representative applications in tissue engineering include imaging the distribution of human mesenchymal stem cells embedded in hydrogels, imaging of bio-printed tissues, and in vivo applications.

Three dimensional mesoscale analysis of translamellar cross‐bridge morphologies in the annulus fibrosus using optical coherence tomography

The defining characteristic of the annulus fibrosus (AF) of the intervertebral disc (IVD) has long been the lamellar structures that consist of highly ordered collagen fibers arranged in alternating oblique angles from one layer to the next. However, a series of recent histologic studies have demonstrated that AF lamellae contain elastin‐ and type VI collagen‐rich secondary “cross‐bridge” structures across lamellae. In this study, we use optical coherence tomography (OCT) to elucidate the three‐dimensional (3‐D) morphologies of these translamellar cross‐bridges in AF tissues. Mesoscale volumetric images by OCT revealed a 3‐D network of heterogeneously distributed cross‐bridges. The results of this study confirm the translamellar cross‐bridge is identifiable as a distinguishable structure, which lies in the interbundle space of adjacent lamellae and crisscrosses multiple lamellae in the radial direction. In contrast to previously proposed models extrapolated from 2–D sections, results from this current study show that translamellar cross‐bridges exist as a complex, interconnected network. We also found much greater variation in lengths of cross‐bridges within the interbundle space of lamellae (0.8–1.4 mm from the current study versus 0.3–0.6 mm from 2–D sections). OCT‐based 3–D morphology of translamellar cross‐bridge provides novel insight into the AF structure.

Depth-resolved imaging of colon tumor using optical coherence tomography and fluorescence laminar optical tomography

Early detection of neoplastic changes remains a critical challenge in clinical cancer diagnosis and treatment. Many cancers arise from epithelial layers such as those of the gastrointestinal (GI) tract. Current standard endoscopic technology is difficult to detect the subsurface lesions. In this research, we investigated the feasibility of a novel multi-modal optical imaging approach including high-resolution optical coherence tomography (OCT) and high-sensitivity fluorescence laminar optical tomography (FLOT) for structural and molecular imaging. The C57BL/6J-ApcMin/J mice were imaged using OCT and FLOT, and the correlated histopathological diagnosis was obtained. Quantitative structural (scattering coefficient) and molecular (relative enzyme activity) parameters were obtained from OCT and FLOT images for multi-parametric analysis. This multi-modal imaging method has demonstrated the feasibility for more accurate diagnosis with 88.23% (82.35%) for sensitivity (specificity) compared to either modality alone. This study suggested that combining OCT and FLOT is promising for subsurface cancer detection, diagnosis, and characterization.

OPTIMIZATION OF DESIGN PARAMETERS FOR FLUORESCENCE LAMINAR OPTICAL TOMOGRAPHY

Laminar optical tomography (LOT) is a mesoscopic tomographic imaging technique ranging between confocal microscopy and diffuse optical tomography (DOT). Fluorescence LOT (FLOT) provides depth-resolved molecular information with 100–200 μm resolution over 2–3 mm depth. In this study, we use Monte Carlo simulation and singular-value analysis (SVA) to optimize the source-detector configurations for potential enhancement of FLOT imaging performance. The effects of different design parameters, including source incidence and detector collection angles, detector number, and sampling density, are presented. The results indicate that angled incidence/detection configuration might improve the imaging resolution and depth sensitivity, especially for low-scattering medium. Increasing the number of detectors and the number of scanning steps will also result in enhanced imaging performance. We also demonstrate that the optimal imaging performance depends upon the background scattering coefficient. Our result might prov...

In Vivo Mesoscopic Voltage-Sensitive Dye Imaging of Brain Activation.

We applied time-resolved angled Fluorescence Laminar Optical Tomography (aFLOT) imaging system to record 3D neural activities evoked in the whisker-barrel system of mice in vivo, in response to single whisker deflection.

Multilayer thin-film phantoms for axial contrast transfer function measurement in optical coherence tomography.

In optical coherence tomography (OCT), axial resolution is one of the most critical parameters impacting image quality. It is commonly measured by determining the point spread function (PSF) based on a specular surface reflection. The contrast transfer function (CTF) provides more insights into an imaging system’s resolving characteristics and can be readily generated in a system-independent manner, without consideration for image pixel size. In this study, we developed a test method for determination of CTF based on multi-layer, thin-film phantoms, evaluated using spectral- and time-domain OCT platforms with different axial resolution values. Phantoms representing six spatial frequencies were fabricated and imaged. The fabrication process involved spin coating silicone films with precise thicknesses in the 8-40 μm range. Alternating layers were doped with a specified concentration of scattering particles. Validation of layer optical properties and thicknesses were achieved with spectrophotometry and stylus profilometry, respectively. OCT B-scans were used to calculate CTFs and results were compared with convetional PSF measurements based on specular reflections. Testing of these phantoms indicated that our approach can provide direct access to axial resolution characteristics highly relevant to image quality. Furthermore, tissue phantoms based on our thin-film fabrication approach may have a wide range of additional applications in optical imaging and spectroscopy.

Anatomy and Cellular Constituents of the Human Olfactory Mucosa: A Review

Studies using animal models have recently suggested that the olfactory mucosa may be a source of cells capable of stimulating and contributing to complex neurologic regeneration. Several groups have already transplanted cell derivatives from the olfactory mucosa into injury models, and the results so far have been promising. To fully appreciate the meaning of these experiments, a better understanding of the cellular biology and physiology of the olfactory system is necessary. It is therefore of utmost importance for us to first identify and understand its constituents.

Variations in optical coherence tomography resolution and uniformity: a multi-system performance comparison

Point spread function (PSF) phantoms based on unstructured distributions of sub-resolution particles in a transparent matrix have been demonstrated as a useful tool for evaluating resolution and its spatial variation across image volumes in optical coherence tomography (OCT) systems. Measurements based on PSF phantoms have the potential to become a standard test method for consistent, objective and quantitative inter-comparison of OCT system performance. Towards this end, we have evaluated three PSF phantoms and investigated their ability to compare the performance of four OCT systems. The phantoms are based on 260-nm-diameter gold nanoshells, 400-nm-diameter iron oxide particles and 1.5-micron-diameter silica particles. The OCT systems included spectral-domain and swept source systems in free-beam geometries as well as a time-domain system in both free-beam and fiberoptic probe geometries. Results indicated that iron oxide particles and gold nanoshells were most effective for measuring spatial variations in the magnitude and shape of PSFs across the image volume. The intensity of individual particles was also used to evaluate spatial variations in signal intensity uniformity. Significant system-to-system differences in resolution and signal intensity and their spatial variation were readily quantified. The phantoms proved useful for identification and characterization of irregularities such as astigmatism. Our multi-system results provide evidence of the practical utility of PSF-phantom-based test methods for quantitative inter-comparison of OCT system resolution and signal uniformity.