Volume 137, Issue 5, Pages 1584-1592.e1 (November 2009) Feasibility and Reliability of Pancreatic Cancer Staging Using Fiberoptic Confocal Fluorescence Microscopy in Rats  Mihaela Ignat, Marc Aprahamian, Veronique Lindner, Anaïs Altmeyer, Silvana Perretta, Bernard Dallemagne, Didier Mutter, Jacques Marescaux  Gastroenterology  Volume 137, Issue 5, Pages 1584-1592.e1 (November 2009) DOI: 10.1053/j.gastro.2009.07.045 Copyright © 2009 AGA Institute Terms and Conditions

Figure 1 Systematic FCFM scanning of lymphatic territories. (A) Scanning of the splenic lymphatic chain draining a 4-week pancreatic tumor (t) and the spleen (s), with fiber optic miniprobe (f) delivering near-infrared laser excitation (660 nm) and collecting fluorescence emission (680–800 nm). (B) Anatomic drawing of the rat pancreatic tail and principal lymph node groups (1) splenic, (2) mesenteric, (3) para-aortic, and (4) celiac trunk. Gastroenterology 2009 137, 1584-1592.e1DOI: (10.1053/j.gastro.2009.07.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 2 Representative patterns of normal pancreas and pancreatic tumors in FCFM; and their corresponding H&E micrograph. FCFM image rendering of normal pancreas: (A) by FCFM; arrowhead indicates a low fluorescent ductal cell and (B) in corresponding H&E micrograph, with arrowhead indicating a normal pancreatic duct. FCFM image rendering of ductal adenocarcinoma: (C) by FCFM; small arrows surround a ductal structure, and arrowhead indicates a large cancer cell and (D) corresponding H&E micrograph, with small arrows indicating ductal structure and arrowhead the large cancer cell. Gastroenterology 2009 137, 1584-1592.e1DOI: (10.1053/j.gastro.2009.07.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 3 Lymph node imaging in FCFM and H&E. Normal lymph node (A) by FCFM with small fluorescent cells forming the fine cortex, surrounding nonfluorescent lymphoid follicles, and (B) corresponding H&E micrograph. Inflammatory lymph node (C) by FCFM with small fluorescent cells scattered in the field (D) and corresponding histiocytosis in H&E micrograph. Metastatic lymph node (E) by FCFM with large fluorescent cells, with increased nuclear/cytoplasm ratio, exhibiting the pathognomonic ductal pattern of pancreatic cancer and (F) corresponding H&E micrograph of a metastatic lymph node. Gastroenterology 2009 137, 1584-1592.e1DOI: (10.1053/j.gastro.2009.07.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 4 FCFM of microscopic peritoneal metastasis. (A) Characteristic pattern of ductal adenocarcinoma in FCFM present in the greater omentum. Arrows indicate the border of the metastasis and (B) corresponding H&E micrograph, which confirmed the diagnosis of a 0.3-mm nodule of carcinomatosis. Gastroenterology 2009 137, 1584-1592.e1DOI: (10.1053/j.gastro.2009.07.045) Copyright © 2009 AGA Institute Terms and Conditions

Supplementary Figure 1 FCFM for primary tumor detection. FCFM was performed in 8 randomized animals, after intrapancreatic injection of either tumor cells (n = 4) or cell carrier medium (n = 4) in early tumor development stage. Presence or absence of tumor could not firmly be assessed by gross examination. The results of such “virtual biopsies” (displayed in the left part of the panels) were blindly confirmed on the corresponding H&E slides of the analyzed areas (right part of the panels). A, B, D, and F correspond to pancreatic ductal adenocarcinoma; C and E to normal pancreas; and G and H to pancreatic edema and hemorrhagic infiltration, respectively. Gastroenterology 2009 137, 1584-1592.e1DOI: (10.1053/j.gastro.2009.07.045) Copyright © 2009 AGA Institute Terms and Conditions