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Peer-reviewedCC BYJournal of translational internal medicine

Rapid, label-free differentiation of benign and malignant gastric lesions using two-photon microscopy and quantitative collagen analysis

  • Bo Ning1,2
  • Xiaoqing Zhou2,3
  • Kaiyue Guan2,3
  • Biao Yan4
  • Qianqian Chen2
  • Jinping Li2
  • Shenzhen Liu2
  • Qun Shao2
  • Lihua Sun2
  • Sikun Zhang2,3
  • Xinhua Ji5
  • Dawei Li4
2 more authors
  • Jinfeng Zang5
  • Enqiang Linghu2,3
5 affiliations
  1. 1Southeast University School of Medicine, Nanjing, Jiangsu Province, China.
  2. 2Department of Gastroenterology, the First Medical Center of PLA General Hospital, Beijing, China.
  3. 3Nankai University School of Medicine, Nankai University, Tianjin, China.
  4. 4The College of Future Technology, Peking University, Beijing, China.
  5. 5Department of Pathology, the First Medical Center of PLA General Hospital, Beijing, China.
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Abstract

Background and objectives

Previous investigations have explored two-photon microscopy (TPM) for differentiating normal and malignant lesions in the digestive tract. The aim of this study is to evaluate label-free TPM for rapid, ex vivo classification of fresh endoscopic human gastric tissues across the spectrum of neoplastic progression, identify quantitative imaging biomarkers, and establish a basis for near real-time diagnostics.Methods

Sixty fresh gastric samples (8 normal, 34 chronic inflammation/metaplasia, 10 atypical hyperplasia, 8 malignant lesions) from 32 patients undergoing routine endoscopic biopsy were imaged using TPM without fixation/staining, generating 60 mosaics and 122 3D images. Key morphological features were established and validated by two blinded pathologists. Normal, chronic inflammation/metaplasia lesions were classified into the low-risk group, while atypical hyperplasia and malignant lesions were assigned to the high-risk group. Second-harmonic generation (SHG) quantified collagen area percentage (Area%) and orientation dispersion (Dispersion) to differentiate low-risk group from high-risk group.Results

TPM provided high-resolution visualization of cellular/stromal features essential for pathology, comparable to histology but significantly faster. Blinded review showed substantial inter-reader agreement (Cohen's κ = 0.793) and 80.3% diagnostic accuracy across five tissue types. Quantitative analysis revealed high-risk lesions had significantly greater collagen Dispersion (U = 684, P < 0.001, r = -0.498) and higher Area% (U = 376, P < 0.001, r = -0.644) than low-risk lesions.Conclusions

TPM offers a robust, label-free, rapid method for ex vivo assessment of gastric biopsies, accurately differentiating lesions from normal to malignant lesions. Increased collagen Dispersion and Area% serve as objective biomarkers for distinguishing high-risk lesions. These findings support TPM's transformative potential for near real-time pathological evaluation, accelerating diagnostics, improving biopsy targeting, and enabling AI-assisted bedside/intraprocedural tools in gastric atypical hyperplasia and malignant lesions management.

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Cite

BibTeXRIS
BibTeX
@article{ning2026rapid,
  title = {{Rapid, label-free differentiation of benign and malignant gastric lesions using two-photon microscopy and quantitative collagen analysis}},
  author = {Bo Ning and Xiaoqing Zhou and Kaiyue Guan and Biao Yan and Qianqian Chen and Jinping Li and Shenzhen Liu and Qun Shao and Lihua Sun and Sikun Zhang and Xinhua Ji and Dawei Li and Jinfeng Zang and Enqiang Linghu},
  journal = {Journal of translational internal medicine},
  year = {2026},
  month = aug,
  doi = {10.1515/jtim-2026-0070},
  url = {https://europepmc.org/article/PMC/PMC13726660},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Rapid, label-free differentiation of benign and malignant gastric lesions using two-photon microscopy and quantitative collagen analysis
AU  - Bo Ning
AU  - Xiaoqing Zhou
AU  - Kaiyue Guan
AU  - Biao Yan
AU  - Qianqian Chen
AU  - Jinping Li
AU  - Shenzhen Liu
AU  - Qun Shao
AU  - Lihua Sun
AU  - Sikun Zhang
AU  - Xinhua Ji
AU  - Dawei Li
AU  - Jinfeng Zang
AU  - Enqiang Linghu
PY  - 2026
DA  - 2026/08/24
JO  - Journal of translational internal medicine
DO  - 10.1515/jtim-2026-0070
UR  - https://europepmc.org/article/PMC/PMC13726660
AB  - Background and objectives  Previous investigations have explored two-photon microscopy (TPM) for differentiating normal and malignant lesions in the digestive tract. The aim of this study is to evaluate label-free TPM for rapid, ex vivo classification of fresh endoscopic human gastric tissues across the spectrum of neoplastic progression, identify quantitative imaging biomarkers, and establish a basis for near real-time diagnostics.Methods  Sixty fresh gastric samples (8 normal, 34 chronic inflammation/metaplasia, 10 atypical hyperplasia, 8 malignant lesions) from 32 patients undergoing routine endoscopic biopsy were imaged using TPM without fixation/staining, generating 60 mosaics and 122 3D images. Key morphological features were established and validated by two blinded pathologists. Normal, chronic inflammation/metaplasia lesions were classified into the low-risk group, while atypical hyperplasia and malignant lesions were assigned to the high-risk group. Second-harmonic generation (SHG) quantified collagen area percentage (Area%) and orientation dispersion (Dispersion) to differentiate low-risk group from high-risk group.Results  TPM provided high-resolution visualization of cellular/stromal features essential for pathology, comparable to histology but significantly faster. Blinded review showed substantial inter-reader agreement (Cohen's κ = 0.793) and 80.3% diagnostic accuracy across five tissue types. Quantitative analysis revealed high-risk lesions had significantly greater collagen Dispersion (U = 684, P < 0.001, r = -0.498) and higher Area% (U = 376, P < 0.001, r = -0.644) than low-risk lesions.Conclusions  TPM offers a robust, label-free, rapid method for ex vivo assessment of gastric biopsies, accurately differentiating lesions from normal to malignant lesions. Increased collagen Dispersion and Area% serve as objective biomarkers for distinguishing high-risk lesions. These findings support TPM's transformative potential for near real-time pathological evaluation, accelerating diagnostics, improving biopsy targeting, and enabling AI-assisted bedside/intraprocedural tools in gastric atypical hyperplasia and malignant lesions management.
SN  - 2450-131X
SN  - 2224-4018
ER  -