A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study
BackgroundEndoscopy is the most important tool for gastric cancer diagnosis. However, it relies on naked-eye evaluation by endoscopists, and the histopathologic confirmation is time-consuming. We aimed to visualize and measure the activity of various enzymes through two-photon microscopy (TPM) using...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2021-08-01
|
| Series: | Frontiers in Oncology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2021.634219/full |
| _version_ | 1830135341773750272 |
|---|---|
| author | Choong-Kyun Noh Chang Su Lim Gil Ho Lee Myung Ki Cho Hyo Won Lee Jin Roh Young Bae Kim Eunyoung Lee Bumhee Park Hwan Myung Kim Sung Jae Shin |
| author_facet | Choong-Kyun Noh Chang Su Lim Gil Ho Lee Myung Ki Cho Hyo Won Lee Jin Roh Young Bae Kim Eunyoung Lee Bumhee Park Hwan Myung Kim Sung Jae Shin |
| author_sort | Choong-Kyun Noh |
| collection | DOAJ |
| description | BackgroundEndoscopy is the most important tool for gastric cancer diagnosis. However, it relies on naked-eye evaluation by endoscopists, and the histopathologic confirmation is time-consuming. We aimed to visualize and measure the activity of various enzymes through two-photon microscopy (TPM) using fluorescent probes and assess its diagnostic potential in gastric cancer.Methodsβ-Galactosidase (β-gal), carboxylesterase (CES), and human NAD(P)H: quinone oxidoreductase (hNQO1) enzyme activities in the normal mucosa, ulcer, adenoma, and gastric cancer biopsy samples were measured using two-photon enzyme probes. The fluorescence emission ratio at long and short wavelengths (Ch2/Ch1) for each probe was comparatively analyzed. Approximately 8,000 – 9,000 sectional images in each group were obtained by measuring the Ch2/Ch1 ratio according to the tissue depth. Each probe was cross-validated by measuring enzymatic activity from a solution containing lysed tissue.ResultsTotal of 76 subjects were enrolled in this pilot study (normal 21, ulcer 18, adenoma 17, and cancer 20 patients, respectively). There were significant differences in the mean ratio values of β-gal (0.656 ± 0.142 vs. 1.127 ± 0.109, P < 0.001) and CES (0.876 ± 0.049 vs. 0.579 ± 0.089, P < 0.001) between the normal and cancer, respectively. The mean ratio value of cancer tissues was different compared to ulcer and adenoma (P < 0.001). The hNQO1 activity showed no significant difference between cancer and other conditions. Normal mucosa and cancer were visually and quantitatively distinguished through β-gal and CES analyses using TPM images, and enzymatic activity according to depth, was determined using sectional TPM ratiometric images. The results obtained from lysis buffer-treated tissue were consistent with TPM results.ConclusionsTPM imaging using ratiometric fluorescent probes enabled the discrimination of gastric cancer from normal, ulcer, and adenoma. This novel method can help in a visual differentiation and provide quantitative depth profiling in gastric cancer diagnosis. |
| first_indexed | 2024-12-17T07:30:05Z |
| format | Article |
| id | doaj.art-9cb4eb5230e64ed38cc829fd6ba41572 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-12-17T07:30:05Z |
| publishDate | 2021-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-9cb4eb5230e64ed38cc829fd6ba415722022-12-21T21:58:31ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2021-08-011110.3389/fonc.2021.634219634219A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot StudyChoong-Kyun Noh0Chang Su Lim1Gil Ho Lee2Myung Ki Cho3Hyo Won Lee4Jin Roh5Young Bae Kim6Eunyoung Lee7Bumhee Park8Hwan Myung Kim9Sung Jae Shin10Department of Gastroenterology, Ajou University School of Medicine, Suwon, South KoreaDepartment of Energy Systems Research and Department of Chemistry, Ajou University, Suwon, South KoreaDepartment of Gastroenterology, Ajou University School of Medicine, Suwon, South KoreaDepartment of Energy Systems Research and Department of Chemistry, Ajou University, Suwon, South KoreaDepartment of Energy Systems Research and Department of Chemistry, Ajou University, Suwon, South KoreaDepartment of Pathology, Ajou University School of Medicine, Suwon, South KoreaDepartment of Pathology, Ajou University School of Medicine, Suwon, South KoreaOffice of Biostatistics, Ajou Research Institute for Innovation Medicine, Ajou University Medical Center, Suwon, South KoreaOffice of Biostatistics, Ajou Research Institute for Innovation Medicine, Ajou University Medical Center, Suwon, South KoreaDepartment of Energy Systems Research and Department of Chemistry, Ajou University, Suwon, South KoreaDepartment of Gastroenterology, Ajou University School of Medicine, Suwon, South KoreaBackgroundEndoscopy is the most important tool for gastric cancer diagnosis. However, it relies on naked-eye evaluation by endoscopists, and the histopathologic confirmation is time-consuming. We aimed to visualize and measure the activity of various enzymes through two-photon microscopy (TPM) using fluorescent probes and assess its diagnostic potential in gastric cancer.Methodsβ-Galactosidase (β-gal), carboxylesterase (CES), and human NAD(P)H: quinone oxidoreductase (hNQO1) enzyme activities in the normal mucosa, ulcer, adenoma, and gastric cancer biopsy samples were measured using two-photon enzyme probes. The fluorescence emission ratio at long and short wavelengths (Ch2/Ch1) for each probe was comparatively analyzed. Approximately 8,000 – 9,000 sectional images in each group were obtained by measuring the Ch2/Ch1 ratio according to the tissue depth. Each probe was cross-validated by measuring enzymatic activity from a solution containing lysed tissue.ResultsTotal of 76 subjects were enrolled in this pilot study (normal 21, ulcer 18, adenoma 17, and cancer 20 patients, respectively). There were significant differences in the mean ratio values of β-gal (0.656 ± 0.142 vs. 1.127 ± 0.109, P < 0.001) and CES (0.876 ± 0.049 vs. 0.579 ± 0.089, P < 0.001) between the normal and cancer, respectively. The mean ratio value of cancer tissues was different compared to ulcer and adenoma (P < 0.001). The hNQO1 activity showed no significant difference between cancer and other conditions. Normal mucosa and cancer were visually and quantitatively distinguished through β-gal and CES analyses using TPM images, and enzymatic activity according to depth, was determined using sectional TPM ratiometric images. The results obtained from lysis buffer-treated tissue were consistent with TPM results.ConclusionsTPM imaging using ratiometric fluorescent probes enabled the discrimination of gastric cancer from normal, ulcer, and adenoma. This novel method can help in a visual differentiation and provide quantitative depth profiling in gastric cancer diagnosis.https://www.frontiersin.org/articles/10.3389/fonc.2021.634219/fullgastric cancerdiagnosisratiometric fluorescent probestwo-photon fluorescent imagingendoscopy |
| spellingShingle | Choong-Kyun Noh Chang Su Lim Gil Ho Lee Myung Ki Cho Hyo Won Lee Jin Roh Young Bae Kim Eunyoung Lee Bumhee Park Hwan Myung Kim Sung Jae Shin A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study Frontiers in Oncology gastric cancer diagnosis ratiometric fluorescent probes two-photon fluorescent imaging endoscopy |
| title | A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study |
| title_full | A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study |
| title_fullStr | A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study |
| title_full_unstemmed | A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study |
| title_short | A Diagnostic Method for Gastric Cancer Using Two-Photon Microscopy With Enzyme-Selective Fluorescent Probes: A Pilot Study |
| title_sort | diagnostic method for gastric cancer using two photon microscopy with enzyme selective fluorescent probes a pilot study |
| topic | gastric cancer diagnosis ratiometric fluorescent probes two-photon fluorescent imaging endoscopy |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2021.634219/full |
| work_keys_str_mv | AT choongkyunnoh adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT changsulim adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT gilholee adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT myungkicho adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT hyowonlee adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT jinroh adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT youngbaekim adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT eunyounglee adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT bumheepark adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT hwanmyungkim adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT sungjaeshin adiagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT choongkyunnoh diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT changsulim diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT gilholee diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT myungkicho diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT hyowonlee diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT jinroh diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT youngbaekim diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT eunyounglee diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT bumheepark diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT hwanmyungkim diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy AT sungjaeshin diagnosticmethodforgastriccancerusingtwophotonmicroscopywithenzymeselectivefluorescentprobesapilotstudy |