H1299-SoNar- and H1299-cpYFP-expressing cells (10×106) suspended in 0.1 ml serum-free RPMI1640 with an equal volume of BD Matrigel Matrix (BD Biosciences-356237) were inoculated s.c. in the right axilla of each mouse. When tumor size reached 0.6-1.2 cm in diameter, mice were anaesthetized intraperitoneally with sodium pentobarbital (100 mg/kg body weight), placed in a holding chamber, and given an intravenous tail vein injection with 100 mM sodium pyruvate (Invitrogen) or 600 μM KP372-1 (Echelon Biosciences Inc.) suspended in 0.9% NaCl in a total volume of 0.3 ml. The dual-excitation ratio imaging in tumor tissues expressing SoNar or cpYFP was obtained by a Kodak Multispectral FX imaging system (Carestream Molecular Imaging) with excitation filters 420 BP 20 nm and 490 BP 20 nm, and an emission filter 535 BP 50 nm (for both excitation wavelengths).
Multispectral fx imaging system
Manufactured by Carestream
The Multispectral FX imaging system is a lab equipment product that captures images using multiple wavelengths of light. It is designed to provide high-quality, detailed images for a variety of applications.
Automatically generated - may contain errors
2 protocols using multispectral fx imaging system
1
In Vivo Imaging of Tumor Metabolic Activity
2
SSTR2 Expression and Ga-DOTATOC Uptake
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A549 and Panc-1 cell lines with different expression levels of GFP (low to high) were seeded in 6-well plates at 5×105 cells per well in triplicate and allowed to grow for 48 h. To quantitate the SSTR2 expression, Western blot assay was performed on the samples as previously described 21 (link). Briefly, whole protein extract purification was performed on the cell sediments. Protein samples (30 µL) were loaded onto SDS-polyacrylamide gels (Bio-Rad) and run at 120 V and 14 mA for 1.5 h. Gels were blotted on polyvinylidene difluoride (PVDF) membrane and the blots incubated overnight at 4 °C with anti-human SSTR2 monoclonal antibody (Abcam) at 1/500 dilution. ß-actin monoclonal antibody (Santa Cruz) at 1/1000 dilution was used as an internal control. Detection was performed using the BM Chemiluminescence Western Blotting Kit (Roche) and imaged on the in vivo Multispectral FX imaging system (Carestream). The SSTR2 expression corrected for ß-actin was correlated to the 68Ga-DOTATOC uptake per cell.
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