Xenogen ivis lumina in vivo imaging system

To check ability of HABN to target the tumor tissue, C57BL/6 mice were inoculated subcutaneously with 3 × 10⁵ MC38 cells on day 0. After treating animals of with 10 mg/kg of HABN-Cy5.5 (having the equivalent mass of free Cy5.5) or 0.25 mg/kg of free Cy5.5 on day 25, whole body fluorescence intensities were monitored at predetermined times using a Xenogen IVIS Lumina in vivo imaging system (PerkinElmer) with a Cy 5.5 filter channel and an exposure time of 5 s. In vivo images were acquired and analyzed using IVIS Lumina Living Image Software (v.4.5.5, PerkinElmer). After 24 h of treating animals of each group, mice were euthanized, and organs including heart, lung, kidney, spleen, liver, and tumor were excised. Fluorescence intensity in organs from each group was analyzed using a Xenogen IVIS Lumina in vivo imaging system (PerkinElmer) with a Cy 5.5 filter channel and an exposure time of 5 s. For the HABN-Cy5.5 uptake studies in vivo, animals were euthanized on day 12, and tumor tissues harvested, followed by flow cytometric analysis.