Abstract
Background While endocytoscopy allows for in vivo cellular-level observation, achieving high-quality staining remains a technical challenge. Aims To develop and validate a simple, reliable scale for assessing and improving endocytoscopic staining quality. Methods We developed a four-dimension staining quality score and validated it on a retrospective image cohort (n = 2692). The scale’s reliability, validity, and internal consistency were assessed, and associated factors were identified using multinomial regression. The scale was then externally validated in a separate retrospective image cohort (n = 2413) to confirm its clinical applicability. Results The scale showed inter-rater agreement (ICC = 0.992) and reliability (Cronbach’s alpha = 0.81), with a single principal factor explaining 62.3% of variance.The Spearman correlation between the gold standard score and AOD value was 0.81. Staining quality was influenced by lesion characteristics and physician experience (P < 0.05). After one month of training, junior physicians showed improved agreement with gold standard scores, with Kappa values rising from 0.61, 0.66, and 0.69 to 0.81, 0.83, and 0.82. Higher staining quality was also significantly associated with better concordance between Endocytoscopy imaging and histopathology (P < 0.001). Conclusion This endocytoscopic staining quality score is a reliable and valid tool to guide technique refinement and improve diagnostic accuracy.
Data availability
Data available on request from the corresponding author.
References
Neumann, H. et al. Advanced colonoscopic imaging using endocytoscopy. Dig. Endoscopy: Official J. Japan Gastroenterological Endoscopy Soc. 27, 232–238 (2015).
Kwon, R. S. et al. Endocytoscopy Gastrointest. Endoscopy 70:610–613 (2009).
Barua, I., Mori, Y. & Bretthauer, M. Colorectal polyp characterization with endocytoscopy: Ready for widespread implementation with artificial intelligence? Best practice & research Clinical gastroenterology 52–53:101721 (2021).
Zhang, H. et al. Diagnostic accuracy of endocytoscopy via artificial intelligence in colorectal lesions: A systematic review and meta–analysis. PLoS One. 18, e0294930 (2023).
Ichimasa, K. et al. Double staining with crystal Violet and methylene blue is appropriate for colonic endocytoscopy: an in vivo prospective pilot study. Dig. Endoscopy: Official J. Japan Gastroenterological Endoscopy Soc. 26, 403–408 (2014).
Kumagai, Y. et al. ,. Endocytoscopic Observation of Esophageal Lesions: Our Own Experience and a Review of the Literature. Diagnostics (Basel, Switzerland) 12 (2022).
Abad M. R. A, Shimamura Y., Fujiyoshi Y., Seewald, S. & Inoue H. Endocytoscopy: technology and clinical application in upper gastrointestinal tract. Transl. Gastroenterol. Hepatol. 5, 28 (2020).
Kodashima, S. et al. Ex-vivo study of high-magnification chromoendoscopy in the Gastrointestinal tract to determine the optimal staining conditions for endocytoscopy. Endoscopy 38, 1115–1121 (2006).
Sasajima, K. et al. Real-time in vivo virtual histology of colorectal lesions when using the endocytoscopy system. Gastrointest. Endosc. 63, 1010–1017 (2006).
Kutsukawa, M. et al. Efficiency of endocytoscopy in differentiating types of serrated polyps. Gastrointest. Endosc. 79, 648–656 (2014).
Kumagai, Y. et al. Endocytoscopic observation of esophageal squamous cell carcinoma. Dig. Endoscopy: Official J. Japan Gastroenterological Endoscopy Soc. 22, 10–16 (2010).
Minsky, B. & Mies, C. The clinical significance of vascular invasion in colorectal cancer. Dis. Colon Rectum. 32, 794–803 (1989).
Sedlak, J. C., Yilmaz, Ö. H. & Roper, J. Metabolism and Colorectal Cancer. Annu. Rev. Pathol. 18, 467–492 (2023).
Majumder, S. et al. Endocytoscopy with automated multispectral intestinal barrier pathology imaging for assessment of deep healing to predict outcomes in ulcerative colitis. Gut 73, 1603–1606 (2024).
Iacucci M., Parigi T. L., Bazarova A. et al. Response to: Correspondence on “PICaSSO Histologic Remission Index (PHRI) in ulcerative colitis: development of a novel simplified histological score for monitoring mucosal healing and predicting clinical outcomes and its applicability in an artificial intelligence system” by Wong et al. Gut 72(4): 807–808 (2023).
Funding
This work was supported by The Ministry of Science and Technology of China (grant number: 2022YFE0118100).
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Conception: Jiawei Fan, He Zhu, Ke Tao, Hong Xu; Data Curation: Mingqing Liu, Fengming Ni, Dong Yang, Guohua Jin, Ke Tao; Supervision: He Zhu, Ke Tao, Hong Xu; Writing – Original Draft Preparation: Jiawei Fan, He Zhu; Writing – Review & Editing: Ke Tao, Hong Xu.
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Jiawei Fan, He Zhu, Mingqing Liu, Fengming Ning, Dong Yang, Guohua Jin, Ke Tao and Hong Xu have no conficts of interest or fnancial ties to disclose.
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Fan, J., Zhu, H., Liu, M. et al. Development and validation of a novel scoring scale for colonic endocytoscopy staining quality. Sci Rep (2026). https://doi.org/10.1038/s41598-026-37406-0
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DOI: https://doi.org/10.1038/s41598-026-37406-0