Date of Graduation

12-2018

Document Type

Dissertation

Degree Name

Doctor of Philosophy in Engineering (PhD)

Degree Level

Graduate

Department

Biomedical Engineering

Advisor

Timothy J. Muldoon

Committee Member

Kyle P. Quinn

Second Committee Member

Yuchun Du

Third Committee Member

David A. Zaharoff

Keywords

Azoxymethane, Fluorescein, Microendoscopy, Oral Cell Cytology, Proflavine, Second Harmonic Generation

Abstract

Advances in imaging technology have led to a variety of available clinical and investigational systems. In this collection of studies, we tested the relevance of morphological image feature quantification on several imaging systems and epithelial tissues. Quantification carries the benefit of creating numerical baselines and thresholds of healthy and abnormal tissues, to potentially aid clinicians in determining a diagnosis, as well as providing researchers with standardized, unbiased results for future dissemination and comparison.

Morphological image features in proflavine stained oral cells were compared qualitatively to traditional Giemsa stained cells, and then we quantified the nuclear to cytoplasm ratio. We determined that quantification of proflavine stained cells matched our hypothesis, as the nuclei in oral carcinoma cells were significantly larger than healthy oral cells.

Proflavine has been used in conjunction with translational fluorescence microendoscopy of the gastrointestinal tract, and we demonstrated the ability of our custom algorithm to accurately (up to 85% sensitivity) extract colorectal crypt area and circularity data, which could minimize the burden of training on clinicians. In addition, we proposed fluorescein as an alternative fluorescent dye, providing comparable crypt area and circularity information.

In order to investigate the morphological changes of crypts via the supporting collagen structures, we adapted our quantification algorithm to analyze crypt area, circularity, and an additional shape parameter in second harmonic generation images of label-free freshly resected murine epithelium. Murine models of colorectal cancer (CRC) were imaged at early and late stages of tumor progression, and we noted significant differences between the Control groups and the late cancer stages, with some differences between early and late stages of CRC progression.

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