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Organ-wide human PanIN and microadenoma analyses: bidirectional exocrine-endocrine relationship in early pancreas remodeling

Authors: Mei-Hsin Chung; Hung-Jen Chien; Shih-Jung Peng; Ya-Hsien Chou; Tsai-Chen Chiang; Hsiu-Pi Chang; Chih-Yuan Lee; +4 Authors

Organ-wide human PanIN and microadenoma analyses: bidirectional exocrine-endocrine relationship in early pancreas remodeling

Abstract

Supplemental Materials Supplemental Figures Suppl. Fig. S1 (related to Fig. 1). Detection and confirmation of duct lesion in human donor pancreas (gallery display). Suppl. Fig. S2 (related to Fig. 3). False positive result in fluorescence imaging of pancreatic microadenoma. (A-C) Gross view and side-by-side display of fluorescence and transmitted light signals of microadenoma. The three images were derived from an optically cleared microadenoma (refractive index: 1.52). A: microadenoma alongside pancreatic lobules (red, blood vessels, CD31; green, lymphatic vessels, D2-40; white, nuclei, DAPI). B: enlarged fluorescence image of vasculature (blood and lymphatic vessels). C: overlay of transmitted light and nuclear signals. Adipocytes and blood clots (black spots) are prominently seen in C via in-depth transmitted light signals. Number 1-6 indicate the adipocytes (#1-2) and blood clots (#3-6) shown in both B and C. Alphabet a-f indicate the adipocytes and blood clots appear only in C. CD31 signals of B is presented in D. B-D examine the same microenvironment. (D, E) False positive result revealed by analysis of CD31 fluorescence signals of microadenoma. Relative signals intensity along the central line of D (0-2,000 pixels) is presented in E. Both CD31-labeled blood vessels and the blood clot-induced autofluorescence create local increases in the signal intensity (E). Number 4 in E indicates the autofluorescence caused by the blood clot #4 in panel B-D. Suppl. Fig. S3 (related to Fig. 4). Detection of PanIN adjacent to microadenoma. (A-C) Microadenoma and its microenvironment visualized via stereomicroscopy. A-C and Fig. 4 examine the same microenvironment (arrow). Vibratome section C is further processed for H&E histology to confirm the PanIN lesion. (D, E) Gross view and enlarged H&E images of PanIN. The gold-standard H&E histology (cyan arrows from C to D to E, same environment) confirms the low-grade PanIN lesion and peri-lesional stroma. Box in D is magnified in E to specify the lesion. =============================================================================================== Supplemental Tables Suppl. Table S1 (Related to Materials and Methods). Summary of primary antibodies used in illustrations. Suppl. Table S2 (Related to Materials and Methods). Summary of color codes presented in illustrations. =============================================================================================== Supplemental Video Suppl. Video S1 (related to Fig. 2). Tile scan and in-depth recording of PanIN microenvironment in human pancreas. This video shows a survey of PanIN microenvironment to identify the associated exocrine-endocrine tissue remodeling. Duct lesion and islet a- and b-cells form a complex in the peri-lobular space. Green, CK7; magenta, glucagon; blue, insulin; white, nuclei. Overlay of fluorescence and transmitted light (gray) signals is used to identify the boundaries of pancreatic lobules and CK7+ epithelium.

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