A Novel Prototype Hyperspectral Microscope Obtains Optical Biopsies and Ascertains Differences between Normal and Carcinomatous Colonic Mucosa

Isaac C Payne, DO1, Paul Rider, MD1, Carole Boudreaux, MD1, Thomas Rich, PhD2, Silas Leavesley, PhD3. 1University of South Alabama Medical Center, 2University of South Alabama – Center for Lung Biology, 3University of South Alabama – Chemical and Biomolecular Engineering

Objective: Conventional white light endoscopy (WLE) is the standard of practice for screening colonoscopy. Over the past decade, implementation of new endoscopic imaging techniques, such as narrow band imaging, chromoendoscopy and endomicroscopy have been utilized to improve detection of mucosal abnormalities during colonoscopy. These techniques have limitations. Utilization of novel microscopic technology capable of performing excitation scanning and acquisition of hyperspectral imaging provides detailed information of colonic mucosal abnormalities.

Description: A hyperspectral, inverted fluorescence microscope (TE2000-U, Nikon Instruments) and prototype tunable excitation filter system were used to achieve fluorescence excitation scanning four different wavelength ranges. Tissue was collected ex vivo for analysis. Matched specimens of normal colon and biopsy proven colorectal adenocarcinoma were analyzed.  Corrected hyperspectral image stacks were constructed. Regions of interest (ROIs) were selected and visualized with four different wavelength bands. Spectrums were extracted and plotted on a graph to be analyzed for differences.

Preliminary Results: Preliminary data indicate normal and biopsy proven adenocarcinoma colonic mucosa have characteristic features that are distinct. Adenocarcinoma tissue samples possess a common spectral shape, with a peak at ~ 400 nm and relative minimum at ~ 430 nm.

Conclusion: Hyperspectral imaging is a novel technique that provides optical biopsies and distinct morphologic and spectral data regarding colonic mucosa. These spectral differences between normal and carcinomatous mucosa are evident with real time imaging. 

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