For near-infrared fluorescence imaging LS174T xenograft bearing mice were co-injected with 50 (0.93 nmol), 100 (1.85 nmol) or 250 µg (4.63 nmol) of both 800CW-AMG211 and 680RD-Mec14. At 24 hours after injection, mice were sacrificed, tumor tissue was harvested, formalin-fixed and paraffin embedded. Four µm sections were incubated for 2 minutes in xylene followed by scanning 800CW-AMG211 and 680RD-Mec14 with Odyssey infrared imaging system (LI-COR Biosciences) for intratumoral distribution. After Odyssey scanning, the same tumor sections were stained with hematoxylin and eosin (H&E). In addition, subsequent tumor slices were stained with immunohistochemistry using 1 µg/mL rabbit monoclonal CEA antibody (11077-R327; Sino Biologicals Inc.). For fluorescent microscopy, an inverted Leica DMI600B fluorescence microscope equipped with a Lumen Dynamics X-Cite 200DC light source was used. Nuclei were stained with Hoechst 33342 (Life Technologies).
Dmi600b fluorescence microscope
Manufactured by Excelitas
The DMI600B is a fluorescence microscope designed for live-cell imaging and high-resolution microscopy. It features a high-intensity, uniform illumination system and advanced optics for producing clear, high-contrast images. The DMI600B is capable of multiple imaging modalities, including brightfield, phase contrast, and fluorescence.
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2 protocols using dmi600b fluorescence microscope
1
Near-infrared Fluorescence Imaging of Xenograft Tumors
2
Targeted Fluorescence Imaging of HT-29 Xenografts
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For NIR fluorescence imaging, mice bearing HT-29 xenografts (n 5 4) were coinjected with 40 mg of 800CW-AMG 110 and 40 mg of 680RD-Mec14. At 24 h after injection mice, were sacrificed and tumor tissue was excised, formalin-fixed, and paraffin-embedded. Tumor slices were stained immunohistochemically for EpCAM (D9S3P; Cell Signaling) and with hematoxylin and eosin. Overview images of intratumoral 680RD-Mec14 and 800CW-AMG 110 distribution were obtained with the Odyssey infrared imaging system (LI-COR Biosciences). For fluorescence microscopy, an inverted Leica DMI600B fluorescence microscope equipped with a Lumen Dynamics X-Cite 200DC light source was used. Nuclei were stained with 49,6-diamidino-2phenylindole (DAPI; Sigma-Aldrich).
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