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. 2020 Aug 4;10:13129. doi: 10.1038/s41598-020-69570-2

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Ce6-PVP-based fluorescence endoscopy improves the detection rate of neoplastic lesions and helps to identify biopsy-worthy areas during colonoscopy in vivo. (a) Representative images displaying an easy-to-miss murine colorectal lesion localized near the anocutaneous line in different imaging modalities. The tumor was missed using white-light endoscopy. Due to its intense fluorescence signal, the tumor was directly detected during fluorescence endoscopy. The post-processed fluorescence signal and the fluorescence/white-light overlay illustrate improved discriminability of the tumor against its surroundings compared to white-light endoscopy alone. H&E staining displays the neoplastic character of the lesion. An experienced pathologist confirmed the diagnosis of high-grade dysplasia based on 3 µm sections of a swiss roll preparation of the colon at variable intervals. Localization relative to the anocutaneous line was used to assure correct matching of histological and endoscopic lesions. (b) Left diagram: Stacked bar chart displaying the percentage of correctly identified neoplastic lesions. The total amount of histologically proven neoplastic lesions is defined as 100% (n = 8). The detection rate is given for white-light endoscopy (75%, 6/8; coloview) and fluorescence endoscopy (100%, 8/8; custom-built device). Right diagram: Scattered dot plot displaying the endoscopic colonic inflammation score [0 to 15]. Bar chart indicates mean (+ /- standard deviation (SD)). (c) Stacked bar chart displaying incorrectly identified lesions as percentage of all identified lesions for white-light, fluorescence, and histology (gold standard test). For white-light endoscopy 45% (5/11 identified lesions) were incorrect in comparison to 81% (34/42 identified lesions) for the fluorescence endoscopy. (d) Native confocal microscopy of Ce6-PVP fluorescence (left) and H&E staining (right) of inflamed colonic mucosa (lower panel) compared to unaltered mucosa (upper panel). An experienced pathologist diagnosed increased proliferation due to tissue regeneration (“reactive focus”) based on representative images of stained 7 µm cryo-sections from a colon segment. (e) Representative images of a false positive fluorescence lesion in different imaging modalities. Consistent with the findings of Fig. 3d, the fluorescence signal identifies a region with strongly altered epithelium. H&E staining displays the irregular character of the lesion. An experienced pathologist diagnosed increased proliferation based on 3 µm sections at intervals of 20 µm from a colon segment sample. Relative topography of the endoscopically detected lesion to the anal canal was used to match the correct colon segment during collection.