Utilizing Mirau-Based Full-Field Optical Coherence Tomography Combined with Fluorescence Microscope on Skin Melanoma Diagnosis
Date Issued
2015
Date
2015
Author(s)
Chou, Ting-Ching
Abstract
Mirau-based full-field optical coherent tomography (Mirau FF-OCT) is a three-dimensional (3D) optical imaging system that has characteristics of non-invasive, no specific demand in preparing the sample and high spatial resolution. It has been tried to be used in clinically dermatology research lately, but part of cases are hard to distinguish lesion correctly. Fluorescence microscopy (FM) uses fluorescence dye to label biological molecule of interest, so it can only present image of specific target. We use immunofluorescence techniques to aid OCT in melanoma diagnosis. We applied labeled antibodies Cyanine Cy™3 to bind to target antibodies MART-1 for melanoma detection. We also obtained large area OCT image with exactly the same specimen, and with the help of dermatologists and pathologists circled the area on the image which showed the presence of melanoma. The H&E stain of the sample is needed as standard contrast. Our research reveals that combining fluorescence microscopy can reduce the case of misidentification dramatically. We did the experiment with two comparison methods. The first method was compared the difference in circled areas by pathologists or dermatologists between large area OCT images and H&E stains image. The second method was compared the difference in detection areas which had been involved by melanoma between combination of OCT with FM and H&E stain. We evaluated the degree of differences by using Cohen''s κ, chi-square test, phi coefficient, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Results showed that dermatologists'' recognition as tumor area varied greatly. κ statistic, a measure of agreement, ranged from 0.74 to disagreement and phi coefficient ranged from 0.77 to 0.22. The results of diagnostic test also showed that some dermatologists recognized tumor areas very well but others did not. The circled areas by pathologists were very close. However, the results from experienced pathologists were not in agreement with H&E stain perfectly and phi coefficient was also less than 0.3. After combining FM, Cohen''s κ, phi coefficient and the results of diagnostic test are greater than 0.9. In this study, by utilizing the combination of Mirau FF-OCT with FM and immunofluorescence techniques, we obtain better results than individual methods in detecting melanoma. The agreement and correlation with H&E stain are all greater than 0.9. These demonstrate that our optical system is very close to H&E stain and has great potential on clinical application in the future.
Subjects
Mirau-based full-field optical coherent tomography (Mirau FF-OCT)
fluorescence microscopy
melanoma
concordance
chi-square test
Type
thesis
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