Measuring the fluorescent quantum efficiency of indocyanine green encapsulated in nanocomposite particulates

We present results of a fluorescent quantum efficiency (Phi(F)) study on the encapsulation of the near-infrared dye indocyanine green (ICG) in bioresorbable calcium phosphate nanoparticles (CPNPs). The Phi(F) (described as the ratio of photons emitted to photons absorbed) provides a quantitative means of describing the fluorescence of an arbitrary molecule. However, standard quantum efficiency measurement techniques provide only the Phi(F) of the smallest fluorescing unit-in the case of a nanoparticle suspension, the nanoparticle itself. This presents a problem in accurately describing the Phi(F) of fluorophores embedded in an inorganic nanoparticle. Combining the incidence of scattering with an evaluation of the differences in local electric field and photochemical environment, we have developed a method to determine the Phi(F) of the constituent fluorescent molecules embedded in such a nanoparticle, which provides a more meaningful comparison with the unencapsulated fluorophore. While applicable to generic systems, we present results obtained by our method for the ICG-CPNP in a phosphate buffered 0.15 M saline solution (PBS, pH 7.4)-specifically, Phi(F), free dye = 0.027 +/- 0.001, Phi(F), particle = 0.053 +/- 0.003, and for the individual encapsulated molecules, Phi(F), molecule = 0.066 +/- 0.004. The method developed also provides insight into the influences of encapsulation and key parameters to engineer resonant enhancement effects from the emission of the encapsulated fluorophores corresponding to an eigenmode of the embedding particle for tailored optical properties.