Hyperspectral Imaging for Detecting Vessel Remodeling in TRPC4 Knockout Rats

Anna AlfordFollow

Department, Center, or Institute

Physics

Presentation Format

Poster

Presentation Type

Off-campus research

Description

HYPERSPECTRAL IMAGING FOR DETECTING VESSEL REMODELING IN TRPC4 KNOCKOUT RATS. Anna Alford. Sponsored by Dr. Silas Leavesley, Thomas Rich, Department of Chemical and Biomolecular Engineering, Department of Pharmacology, Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL. Hyperspectral imaging with fluorescence excitation scanning (HIFEX) is a revolutionary new imaging technique that has been shown to provide increased acquisition speed, sensitivity, and specificity than other comparable methods. By exciting tissue with a sequence of specific wavelengths, HIFEX is able to collect a broad emission spectrum from the tissue that can be used to separate fluorescence signal sources. Recent research has utilized HIFEX in examining differences in lesional and nonlesional tissue form colon cancer patients. HIFEX has so far been able to determine differences in the spectral signatures of healthy and cancerous colon samples, but further research is required to define the exact nature of these differences and potentially enable the future utilization of HIFEX in aiding physicians for cancer detection and treatment. Currently, trial research is being conducted to determine if HIFEX could be used to differentiate tissue between TRPC4 Knockout (KO), Wild Type (WT), and Pulmonary Arterial Hypertensive (PAH) rats. Sections of the aorta and descending aorta were cleaned of surrounding tissue, cut longitudinally, mounted with endothelium facing down, and imaged using a custom inverted fluorescence microscope with a 300 W Xe arc lamp light source and thin film tunable filter array (VF-5, Sutter Instruments) for excitation wavelength selection. The tissue was scanned with wavelengths from 360 nm to 550 nm in 5 nm increments. Resultant images were corrected for background interference using custom Matlab scripts. Tissue-encompassing regions of interest were then selected with ENVI software and the average spectrum of each region was extracted using linear spectral unmixing. Preliminary data indicates that normotensive rats, regardless of whether KO or WT, share a close spectral pattern. The spectra of some of the PAH rats, on the other hand, contain peaks that are not present in the normotensive rats, indicating that a result of PAH could be increasing the quantity of a particular autofluorescent molecule or combination of molecules in the tissue. Further analysis will be required to determine the identification of specific fluorophores and the relationship to PAH-associated remodeling. The results of this trial research and future research using HIFEX hope to develop a detection algorithm that will be able to determine the individual components responsible for a specific part of the excitation spectrum and the relationship of spectral changes to vessel remodeling. This will ultimately aid the future work of researchers and physicians in utilizing HIFEX for faster and more efficient detections of abnormalities in tissue.

Session Number

4

Start Date and Time

4-9-2019 3:00 PM

Location

PAC Gym

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Apr 9th, 3:00 PM

Hyperspectral Imaging for Detecting Vessel Remodeling in TRPC4 Knockout Rats

PAC Gym

HYPERSPECTRAL IMAGING FOR DETECTING VESSEL REMODELING IN TRPC4 KNOCKOUT RATS. Anna Alford. Sponsored by Dr. Silas Leavesley, Thomas Rich, Department of Chemical and Biomolecular Engineering, Department of Pharmacology, Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL. Hyperspectral imaging with fluorescence excitation scanning (HIFEX) is a revolutionary new imaging technique that has been shown to provide increased acquisition speed, sensitivity, and specificity than other comparable methods. By exciting tissue with a sequence of specific wavelengths, HIFEX is able to collect a broad emission spectrum from the tissue that can be used to separate fluorescence signal sources. Recent research has utilized HIFEX in examining differences in lesional and nonlesional tissue form colon cancer patients. HIFEX has so far been able to determine differences in the spectral signatures of healthy and cancerous colon samples, but further research is required to define the exact nature of these differences and potentially enable the future utilization of HIFEX in aiding physicians for cancer detection and treatment. Currently, trial research is being conducted to determine if HIFEX could be used to differentiate tissue between TRPC4 Knockout (KO), Wild Type (WT), and Pulmonary Arterial Hypertensive (PAH) rats. Sections of the aorta and descending aorta were cleaned of surrounding tissue, cut longitudinally, mounted with endothelium facing down, and imaged using a custom inverted fluorescence microscope with a 300 W Xe arc lamp light source and thin film tunable filter array (VF-5, Sutter Instruments) for excitation wavelength selection. The tissue was scanned with wavelengths from 360 nm to 550 nm in 5 nm increments. Resultant images were corrected for background interference using custom Matlab scripts. Tissue-encompassing regions of interest were then selected with ENVI software and the average spectrum of each region was extracted using linear spectral unmixing. Preliminary data indicates that normotensive rats, regardless of whether KO or WT, share a close spectral pattern. The spectra of some of the PAH rats, on the other hand, contain peaks that are not present in the normotensive rats, indicating that a result of PAH could be increasing the quantity of a particular autofluorescent molecule or combination of molecules in the tissue. Further analysis will be required to determine the identification of specific fluorophores and the relationship to PAH-associated remodeling. The results of this trial research and future research using HIFEX hope to develop a detection algorithm that will be able to determine the individual components responsible for a specific part of the excitation spectrum and the relationship of spectral changes to vessel remodeling. This will ultimately aid the future work of researchers and physicians in utilizing HIFEX for faster and more efficient detections of abnormalities in tissue.