Biodistribution studies of nanoparticles using fluorescence imaging: a qualitative or quantitative method?

The biodistribution of Lipid/Calcium/Phosphate (LCP) nanoparticles (NPs) in tumor-bearing mice was investigated using fluorescence imaging. A quantitative validation of this method was done by (3)H and (111)In labeling of the nanoparticles. The biodistribution of LCP NPs containing oligonucleotides was investigated using three different probes: Texas-Red labeled oligonucleotides, (3)H-labeled oligonucleotides, and (111)In-labled calcium phosphate. A discrepancy was found between the radioactivity and the fluorescence signals. Signals from (3)H and (111)In exhibited very similar distribution patterns, suggesting that liver and spleen were the major accumulation sites. However, fluorescence imaging indicated that tumor accumulation was predominant. We further confirmed that the fluorescence signals in both liver and spleen were greatly attenuated compared with those in the tumor due to the intrinsic tissue absorption and scattering. Near-infrared (NIR) dye Cy5.5 also suffered from the same problem, in that the quantitative data from whole organs was dramatically affected by absorption and scattering properties of the tissue. Careful attention must be paid to the quantification and interpretation of fluorescence imaging measurements when comparing different tissues.