Demonstration of metastatic colon cancer cells adjacent to hepatic tissue.

<p>H&E, confocal and fluorescence images of adjacent tissue samples are shown (Confocal and fluorescence images from same slide). The tumor margin was much better delineated when employing fluorescence microscopy.</p

Imaging of IGF-1R targeting of liver metastasis.

<p>Anti-IGF-1R conjugated to PEGylated 650 nm dye selectively labeled the HCT 116-GFP metastatic tumors. Both in vivo (A) and ex vivo (B) imaging show that the 650 nm fluorophore-conjugated IGF-1R antibodies co-localized with HCT 116-GFP fluorescence and more accurately demarked the tumor compared to bright light imaging (white arrowheads). H & E staining of tissue sample (x200) expressing fluorescence confirms the presence of metastatic tumor in the mouse liver.</p

Liver tissue concentrations of the dye anti-CEA antibody conjugates over time.

<p>The liver was assayed to demonstrate tissue accumulation of the non-PEGylated and PEGylated 650 and 750 dyes that were conjugated to chimeric anti-CEA antibody. The non-PEGylated 650 (panel A) and 750 dye antibody conjugates (panel B) accumulated to significantly higher levels in the liver compared to their respective PEGylated dye antibody conjugates.</p

Imaging of MUC1 targeting of orthotopically-transplanted Panc-1 and BxPC-3 pancreatic tumor in vivo.

<p>Fluorescence signals from pancreatic tumors orthotopically transplanted at the tail of the pancreas were detected. Other than the tumor, fluorescence signal was detected from the skin and, bladder and intestinal contents but at lower intensity than the tumor. White arrows indicate pancreatic tumor.</p

Characterization of colon cancer cell lines.

<p>(A) Western blot analysis shows pro-IGF-1R and IGF-1Rα expression at 200 and 130 kDa in colon cancer cell lines, respectively (HT-29 and HCT 116). (B) Labeling of live HCT 116 and HT-29 cells with 550 nm fluorophore-conjugated antibodies shows multiple fluorescent foci on the surface. Representative fluorescence images merged with corresponding DIC (differential interference contrast) images (x60 water immersion objective with the FV1000, using the 559 nm laser).</p

Imaging of MUC1 targeting of pancreatic tumors growing on skin flaps.

<p>Representative images were obtained under white light, and 473 nm and 559 nm lasers on the OV100, and merged. GFP signal from the individual cancer cells and red fluorescent signal from the DyLight 550 fluorophore-conjugated anti-MUC1 antibody at the outside margin of the colony and space between the cancer cells were observed.</p

Characterization of pancreatic cancer cell lines.

<p>(A) Western blot analysis shows MUC1 expression in pancreatic cancer cell lines (BxPC-3 and Panc-1). (B) Flow cytometric analysis shows the expression of MUC1 on the surface of BxPC-3 and Panc-1 cell lines. (C) Immunocytochemistry on live cells shows multiple fluorescent dots on the surface of Panc-1 cells. Representative fluorescence images merged with corresponding DIC (differential interference contrast) images (x60 water immersion objective on FV1000, using the 559 nm laser).</p

Schematic comparison of anti-CEA chimeric antibodies with PEGylated and non-PEGylated NIR dyes.

<p>Four mPEG200 chains are covalently linked to each dye molecule, 4 of which are covalently linked to the chimeric anti-CEA antibody by amide bonds. The mPEG chains alter tissue biodistribution, allowing brighter liver metastases labeling and decreased accumulation in normal organs, particularly the liver.</p

Imaging of IGF-1R targeting of orthotopically-transplanted HCT 116 colon tumors in vivo.

<p>Fluorescence from orthotopically transplanted colon tumors at the cecum was detected before and after abdominal laparotomy. Also, fluorescence from the resected tumor was detected. Weak fluorescence was also detected from the skin and, bladder and intestinal contents, but at much lower intensity than the tumor. White arrows indicate colon tumor.</p

Specificity of anti-CEA antibody conjugated dyes as compared to free dyes.

<p>In all images from A to D, the top panel of images is taken with the liver in anatomical position and the bottom panel is taken with the liver flipped upward to enable visualization of the undersurface. Image A is taken 24-anti-CEA antibody conjugate. Imaging at 750 nm clearly delineates all the HT-29-GFP liver metastases which are not completely visible on imaging with brightfield or GFP imaging (see arrows). Image B is taken 24 hours after injection of the 750 PEG dye only. No visible tumor labeling is noted, with little to no visualization of the dye throughout the animal, despite the GFP signal of the tumor being clearly detected. Image C is taken 24 hours after injection of the 650 PEG-anti-CEA antibody conjugate. There is very precise and clear labeling of tumor that is detected at 650 nm, which correlates well with the GFP signal. With the liver in the anatomical position, the gallbladder is also noted emerging from the underside of the liver. On the underside of the liver, a tumor is detected that is not visible on either GFP or brightfield imaging (see arrows). Image D is taken 24 hours after injection of the 650PEG free dye. There is non-specific distribution of the dye noted at 650 nm, with no selectivity for the tumor despite there being a robust GFP signal.</p