In vivo long-term investigation of tumor bearing mKate2 by an in-house fluorescence molecular imaging system

Abstract

BackgroundOptical imaging is one of the most common, low-cost imaging tools used for investigating the tumor biological behavior in vivo. This study explores the feasibility and sensitivity of a near infrared fluorescent protein mKate2 for a long-term non-invasive tumor imaging in BALB/c nude mice, by using a low-power optical imaging system.MethodsIn this study, breast cancer cell line MDA-MB-435s expressing mKate2 and MDA-MB-231 expressing a dual reporter gene firefly luciferase (fLuc)-GFP were used as cell models. Tumor cells were implanted in different animal body compartments including subcutaneous, abdominal and deep tissue area and closely monitored in real-time. A simple and low-power optical imaging system was set up to image both fluorescence and bioluminescence in live animals.ResultsThe presence of malignant tissue was further confirmed by histopathological assay. Considering its lower exposure time and no need of substrate injection, mKate2 is considered a superior choice for subcutaneous imaging compared with fLuc. On the contrary, fLuc has shown to be a better option when monitoring the tumor in a diffusive area such as abdominal cavity. Furthermore, both reporter genes have shown good stability and sensitivity for deep tissue imaging, i.e. tumor within the liver. In addition, fLuc has shown to be an excellent method for detecting tumor cells in the lung.ConclusionsThe combination of mKate2 and fLuc offers a superior choice for long-term non-invasive real-time investigation of tumor biological behavior in vivo.

A genetic model for in vivo proximity labeling of the mammalian secretome

“…mKate2 encodes a monomeric, photostable, pH resistant, low toxicity, bright far-red fluorescent protein designed as a cellular indicator of cells producing BirA*G3 and activating biotinylation of secretory pathway proteins on biotin administration (Fig. 1B) [20][21][22] . CRE recombination at loxP sites flanking the GFP cassette enables the tissue and time-dependent expression of BirA*G3 and mKate2 13 .…”

The fluorescence mKate2 labeling as a visualizing system for monitoring small extracellular vesicles

“…Zhou et al.’s study demonstrated that combination of mKate2 and firefly luciferase (fLuc) offered a superior choice for long‐term noninvasive real‐time investigation of tumor biological behavior in in BALB/c nude mice. [ 34 ] Other study from Vuletic et al. developed a breast cancer cell line that stably expresses the mKate2 could be as a highly valuable cell model for studying breast cancer detection and anticancer drug evaluation by noninvasive fluorescence imaging in mice.…”

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention

“…Small cell numbers were su cient to monitor the FusionRed labeled cells in the whole body imager [59]. For in vivo imaging studies a cell number from 1 × 10 6 cells per tumor is detectable with this setting already in early tumor stages [60]. Red uorescent proteins showed high sensitivity in subcutaneous, abdominal as well as deep tissue cell implantation.…”