Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices
IntroductionMelanoma diagnosis traditionally relies on microscopic examination of hematoxylin and eosin (H&E) slides by dermatopathologists to search for specific architectural and cytological features. Unfortunately, no single molecular marker exists to reliably differentiate melanoma from...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-11-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2023.1296826/full |
| _version_ | 1797387545887113216 |
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| author | Bruno Schuty Sofía Martínez Sofía Martínez Analía Guerra Federico Lecumberry Julio Magliano Leonel Malacrida Leonel Malacrida |
| author_facet | Bruno Schuty Sofía Martínez Sofía Martínez Analía Guerra Federico Lecumberry Julio Magliano Leonel Malacrida Leonel Malacrida |
| author_sort | Bruno Schuty |
| collection | DOAJ |
| description | IntroductionMelanoma diagnosis traditionally relies on microscopic examination of hematoxylin and eosin (H&E) slides by dermatopathologists to search for specific architectural and cytological features. Unfortunately, no single molecular marker exists to reliably differentiate melanoma from benign lesions such as nevi. This study explored the potential of autofluorescent molecules within tissues to provide molecular fingerprints indicative of degenerated melanocytes in melanoma.MethodsUsing hyperspectral imaging (HSI) and spectral phasor analysis, we investigated autofluorescence patterns in melanoma compared to intradermal nevi. Using UV excitation and a commercial spectral confocal microscope, we acquired label-free HSI data from the whole-slice samples. ResultsOur findings revealed distinct spectral phasor distributions between melanoma and intradermal nevi, with melanoma displaying a broader phasor phase distribution, signifying a more heterogeneous autofluorescence pattern. Notably, longer wavelengths associated with larger phases correlated with regions identified as melanoma by expert dermatopathologists using H&E staining. Quantitative analysis of phase and modulation histograms within the phasor clusters of five melanomas (with Breslow thicknesses ranging from 0.5 mm to 6 mm) and five intradermal nevi consistently highlighted differences between the two groups. We further demonstrated the potential for the discrimination of several melanocytic lesions using center-of-mass comparisons of phase and modulation variables. Remarkably, modulation versus phase center of mass comparisons revealed strong statistical significance among the groups. Additionally, we identified the molecular endogenous markers responsible for tissue autofluorescence, including collagen, elastin, NADH, FAD, and melanin. In melanoma, autofluorescence is characterized by a higher phase contribution, indicating an increase in FAD and melanin in melanocyte nests. In contrast, NADH, elastin, and collagen dominate the autofluorescence of the nevus.DiscussionThis work underscores the potential of autofluorescence and HSI-phasor analysis as valuable tools for quantifying tissue molecular fingerprints, thereby supporting more effective and quantitative melanoma diagnosis. |
| first_indexed | 2024-03-08T22:26:38Z |
| format | Article |
| id | doaj.art-aeccbee9e9444008903907006ffdead4 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-03-08T22:26:38Z |
| publishDate | 2023-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-aeccbee9e9444008903907006ffdead42023-12-18T10:35:32ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2023-11-011310.3389/fonc.2023.12968261296826Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slicesBruno Schuty0Sofía Martínez1Sofía Martínez2Analía Guerra3Federico Lecumberry4Julio Magliano5Leonel Malacrida6Leonel Malacrida7Unidad de Bioimagenología Avanzada, Institut Pasteur de Montevideo, Hospital de Clínicas Universidad de la República, Montevideo, UruguayUnidad de Bioimagenología Avanzada, Institut Pasteur de Montevideo, Hospital de Clínicas Universidad de la República, Montevideo, UruguayUnidad Academica de Dermatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, UruguayUnidad Academica de Dermatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, UruguayInstituto de Ingeniería Eléctrica, Facultad de Ingeniería, Universidad de la República, Montevideo, UruguayUnidad Academica de Dermatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, UruguayUnidad de Bioimagenología Avanzada, Institut Pasteur de Montevideo, Hospital de Clínicas Universidad de la República, Montevideo, UruguayUnidad Academica de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, UruguayIntroductionMelanoma diagnosis traditionally relies on microscopic examination of hematoxylin and eosin (H&E) slides by dermatopathologists to search for specific architectural and cytological features. Unfortunately, no single molecular marker exists to reliably differentiate melanoma from benign lesions such as nevi. This study explored the potential of autofluorescent molecules within tissues to provide molecular fingerprints indicative of degenerated melanocytes in melanoma.MethodsUsing hyperspectral imaging (HSI) and spectral phasor analysis, we investigated autofluorescence patterns in melanoma compared to intradermal nevi. Using UV excitation and a commercial spectral confocal microscope, we acquired label-free HSI data from the whole-slice samples. ResultsOur findings revealed distinct spectral phasor distributions between melanoma and intradermal nevi, with melanoma displaying a broader phasor phase distribution, signifying a more heterogeneous autofluorescence pattern. Notably, longer wavelengths associated with larger phases correlated with regions identified as melanoma by expert dermatopathologists using H&E staining. Quantitative analysis of phase and modulation histograms within the phasor clusters of five melanomas (with Breslow thicknesses ranging from 0.5 mm to 6 mm) and five intradermal nevi consistently highlighted differences between the two groups. We further demonstrated the potential for the discrimination of several melanocytic lesions using center-of-mass comparisons of phase and modulation variables. Remarkably, modulation versus phase center of mass comparisons revealed strong statistical significance among the groups. Additionally, we identified the molecular endogenous markers responsible for tissue autofluorescence, including collagen, elastin, NADH, FAD, and melanin. In melanoma, autofluorescence is characterized by a higher phase contribution, indicating an increase in FAD and melanin in melanocyte nests. In contrast, NADH, elastin, and collagen dominate the autofluorescence of the nevus.DiscussionThis work underscores the potential of autofluorescence and HSI-phasor analysis as valuable tools for quantifying tissue molecular fingerprints, thereby supporting more effective and quantitative melanoma diagnosis.https://www.frontiersin.org/articles/10.3389/fonc.2023.1296826/fullskin cancermelanomafluorescence microscopyhyperspectral imagingphasor analysisspectral phasor |
| spellingShingle | Bruno Schuty Sofía Martínez Sofía Martínez Analía Guerra Federico Lecumberry Julio Magliano Leonel Malacrida Leonel Malacrida Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices Frontiers in Oncology skin cancer melanoma fluorescence microscopy hyperspectral imaging phasor analysis spectral phasor |
| title | Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices |
| title_full | Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices |
| title_fullStr | Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices |
| title_full_unstemmed | Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices |
| title_short | Quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label-free slices |
| title_sort | quantitative melanoma diagnosis using spectral phasor analysis of hyperspectral imaging from label free slices |
| topic | skin cancer melanoma fluorescence microscopy hyperspectral imaging phasor analysis spectral phasor |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2023.1296826/full |
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