Cy5.5 NHS ester (non-sulfonated)

Cell lines

U87MG

Preparation Method

U87MG cells (1 × 10⁵) were cultured for 24 hours, then the cells were washed with PBS and then incubated at 37 °C in the presence of 100 nM Cy5.5-NHS ester for 10 minutes. After the incubation period, cells were washed three times with ice- cold PBS.

Reaction Conditions

The fluorescence signal of the cells was recorded using an Axiovert 200M fluorescence microscope equipped with a Cy5.5 filter set.

Applications

Cy5.5-NHS ester could be used for fluorescence microscopy imaging study with tumor cell lines such as U87MG, C6, A375M, B16F10 and MDAMB-435, as well as for examining the effect of D-glucose or L-glucose on 2-NBDG accumulation.

Animal models

Female athymic nude mice (nu/nu), 4–6 weeks

Preparation Method

Mice were subcutaneously injected with 5 × 10⁶ U87MG glioblastoma cells suspended in 100 μL of PBS. When the tumors reached 0.4–0.6 cm in diameter, the tumor bearing mice were then injected via tail vein with Cy5.5-NHS ester and subjected to optical imaging at various time points postinjection (pi).

Dosage form

0.5 nmol

Applications

Cy5.5-NHS ester could be took advantage of tumor targeting, and the tumor/muscle contrast was clearly visible as early as 30 min postinjection. The highest U87MG tumor/muscle ratio of 3.34 ± 0.23 were achieved 24 hours post-injection for Cy5.5-NHS.

References:

[1]. Zhen C. et al. Near-Infrared Fluorescent Deoxyglucose Analog for Tumor Optical Imaging in Cell Culture and in Living Mice. Bioconjug Chem. 2006 ; 17(3): 662–669.