Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy
Thin tissue sections of normal and tumorous pancreatic tissues stained with hematoxylin and eosin were investigated using multiphoton excitation fluorescence (MPF), second harmonic generation (SHG), and third harmonic generation (THG) microscopies. The cytoplasm, connective tissue, collagen and extr...
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Frontiers Media S.A.
2019-04-01
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Series: | Frontiers in Oncology |
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Online Access: | https://www.frontiersin.org/article/10.3389/fonc.2019.00272/full |
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doaj-9985ffa6febb4b388f4cd638768f5fef |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Danielle Tokarz Richard Cisek Ariana Joseph Ahmad Golaraei Ahmad Golaraei Ahmad Golaraei Kamdin Mirsanaye Kamdin Mirsanaye Serguei Krouglov Serguei Krouglov Sylvia L. Asa Brian C. Wilson Brian C. Wilson Virginijus Barzda Virginijus Barzda |
spellingShingle |
Danielle Tokarz Richard Cisek Ariana Joseph Ahmad Golaraei Ahmad Golaraei Ahmad Golaraei Kamdin Mirsanaye Kamdin Mirsanaye Serguei Krouglov Serguei Krouglov Sylvia L. Asa Brian C. Wilson Brian C. Wilson Virginijus Barzda Virginijus Barzda Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy Frontiers in Oncology collagen optical pathology medical imaging non-linear optical polarimetry non-linear optical microscopy |
author_facet |
Danielle Tokarz Richard Cisek Ariana Joseph Ahmad Golaraei Ahmad Golaraei Ahmad Golaraei Kamdin Mirsanaye Kamdin Mirsanaye Serguei Krouglov Serguei Krouglov Sylvia L. Asa Brian C. Wilson Brian C. Wilson Virginijus Barzda Virginijus Barzda |
author_sort |
Danielle Tokarz |
title |
Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy |
title_short |
Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy |
title_full |
Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy |
title_fullStr |
Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy |
title_full_unstemmed |
Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation Microscopy |
title_sort |
characterization of pancreatic cancer tissue using multiphoton excitation fluorescence and polarization-sensitive harmonic generation microscopy |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Oncology |
issn |
2234-943X |
publishDate |
2019-04-01 |
description |
Thin tissue sections of normal and tumorous pancreatic tissues stained with hematoxylin and eosin were investigated using multiphoton excitation fluorescence (MPF), second harmonic generation (SHG), and third harmonic generation (THG) microscopies. The cytoplasm, connective tissue, collagen and extracellular structures are visualized with MPF due to the eosin stain, whereas collagen is imaged with endogenous SHG contrast that does not require staining. Cellular structures, including membranous interfaces and nuclear components, are seen with THG due to the aggregation of hematoxylin dye. Changes in the collagen ultrastructure in pancreatic cancer were investigated by a polarization-sensitive SHG microscopy technique, polarization-in, polarization-out (PIPO) SHG. This involves measuring the orientation of the linear polarization of the SHG signal as a function of the linear polarization orientation of the incident laser radiation. From the PIPO SHG data, the second-order non-linear optical susceptibility ratio, χ(2)zzz'/χ(2)zxx', was obtained that serves as a structural parameter for characterizing the tissue. Furthermore, by assuming C6 symmetry, an additional second-order non-linear optical susceptibility ratio, χ(2)xyz'/χ(2)zxx', was obtained, which is a measure of the chirality of the collagen fibers. Statistically-significant differences in the χ(2)zzz'/χ(2)zxx' values were found between tumor and normal pancreatic tissues in periductal, lobular, and parenchymal regions, whereas statistically-significant differences in the full width at half maximum (FWHM) of χ(2)xyz'/χ(2)zxx' occurrence histograms were found between tumor and normal pancreatic tissues in periductal and parenchymal regions. Additionally, the PIPO SHG data were used to determine the degree of linear polarization (DOLP) of the SHG signal, which indicates the relative linear depolarization of the signal. Statistically-significant differences in DOLP values were found between tumor and normal pancreatic tissues in periductal and parenchymal regions. Hence, the differences observed in the χ(2)zzz'/χ(2)zxx' values, the FWHM of χ(2)xyz'/χ(2)zxx' values and the DOLP values could potentially be used to aid pathologists in diagnosing pancreatic cancer. |
topic |
collagen optical pathology medical imaging non-linear optical polarimetry non-linear optical microscopy |
url |
https://www.frontiersin.org/article/10.3389/fonc.2019.00272/full |
work_keys_str_mv |
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doaj-9985ffa6febb4b388f4cd638768f5fef2020-11-24T21:46:31ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2019-04-01910.3389/fonc.2019.00272438974Characterization of Pancreatic Cancer Tissue Using Multiphoton Excitation Fluorescence and Polarization-Sensitive Harmonic Generation MicroscopyDanielle Tokarz0Richard Cisek1Ariana Joseph2Ahmad Golaraei3Ahmad Golaraei4Ahmad Golaraei5Kamdin Mirsanaye6Kamdin Mirsanaye7Serguei Krouglov8Serguei Krouglov9Sylvia L. Asa10Brian C. Wilson11Brian C. Wilson12Virginijus Barzda13Virginijus Barzda14Department of Chemistry, Saint Mary's University, Halifax, NS, CanadaDepartment of Chemistry, Saint Mary's University, Halifax, NS, CanadaDepartment of Chemistry, Saint Mary's University, Halifax, NS, CanadaDepartment of Physics, University of Toronto, Toronto, ON, CanadaDepartment of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, ON, CanadaPrincess Margaret Cancer Centre, University of Toronto, Toronto, ON, CanadaDepartment of Physics, University of Toronto, Toronto, ON, CanadaDepartment of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, ON, CanadaDepartment of Physics, University of Toronto, Toronto, ON, CanadaDepartment of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, ON, CanadaUniversity Health Network, University of Toronto, Toronto, ON, CanadaPrincess Margaret Cancer Centre, University of Toronto, Toronto, ON, CanadaDepartment of Medical Biophysics, University of Toronto, Toronto, ON, CanadaDepartment of Physics, University of Toronto, Toronto, ON, CanadaDepartment of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, ON, CanadaThin tissue sections of normal and tumorous pancreatic tissues stained with hematoxylin and eosin were investigated using multiphoton excitation fluorescence (MPF), second harmonic generation (SHG), and third harmonic generation (THG) microscopies. The cytoplasm, connective tissue, collagen and extracellular structures are visualized with MPF due to the eosin stain, whereas collagen is imaged with endogenous SHG contrast that does not require staining. Cellular structures, including membranous interfaces and nuclear components, are seen with THG due to the aggregation of hematoxylin dye. Changes in the collagen ultrastructure in pancreatic cancer were investigated by a polarization-sensitive SHG microscopy technique, polarization-in, polarization-out (PIPO) SHG. This involves measuring the orientation of the linear polarization of the SHG signal as a function of the linear polarization orientation of the incident laser radiation. From the PIPO SHG data, the second-order non-linear optical susceptibility ratio, χ(2)zzz'/χ(2)zxx', was obtained that serves as a structural parameter for characterizing the tissue. Furthermore, by assuming C6 symmetry, an additional second-order non-linear optical susceptibility ratio, χ(2)xyz'/χ(2)zxx', was obtained, which is a measure of the chirality of the collagen fibers. Statistically-significant differences in the χ(2)zzz'/χ(2)zxx' values were found between tumor and normal pancreatic tissues in periductal, lobular, and parenchymal regions, whereas statistically-significant differences in the full width at half maximum (FWHM) of χ(2)xyz'/χ(2)zxx' occurrence histograms were found between tumor and normal pancreatic tissues in periductal and parenchymal regions. Additionally, the PIPO SHG data were used to determine the degree of linear polarization (DOLP) of the SHG signal, which indicates the relative linear depolarization of the signal. Statistically-significant differences in DOLP values were found between tumor and normal pancreatic tissues in periductal and parenchymal regions. Hence, the differences observed in the χ(2)zzz'/χ(2)zxx' values, the FWHM of χ(2)xyz'/χ(2)zxx' values and the DOLP values could potentially be used to aid pathologists in diagnosing pancreatic cancer.https://www.frontiersin.org/article/10.3389/fonc.2019.00272/fullcollagenoptical pathologymedical imagingnon-linear optical polarimetrynon-linear optical microscopy |