NIR laser scanning microscopy for photophysical characterization of upconversion nanoparticles and nanohybrids

Photophysical characterization of upconversion nanoparticles (UCNPs) and nanohybrids (UCNHs) is more challenging than that of down-conversion nanomaterials. Moreover, it is still difficult to gain knowledge about the homogeneity of the sample and colocalization of emissive chromophores and nanoparticles in nanohybrids. Near infrared laser scanning microscopy (NIR-LSM) is a well-known and useful imaging technique, which enables excitation in the NIR region and has been extensively applied to optical fluorescence imaging of organic fluorophores and nanomaterials, such as quantum dots, which exhibit a short-lived emission. NIR-LSM has recently been used to determine the empirical emission lifetime of UCNPs, thus extending its application range to nanomaterials with a long lifetime emission. Here, we review our previous findings and include new measurements and samples to fully address the potential of this technique. NIR-LSM has proved to be extraordinarily useful not only for photophysical characterization of UCNHs consisting of UCNPs capped with a fluorophore to easily visualize the occurrence of the resonance energy transfer process between the UCNH constituents and their homogeneity, but also to assess the colocalization of the fluorophore and the UCNP in the UCNH; all this information can be acquired on the micro-/nano-meter scale by just taking one image. NIR laser scanning microscopy enables the determination of the emission lifetime of upconversion nanoparticles (UCNPs) and UCNP nanohybrids (UCNHs), and the evaluation of the homogeneity in terms of colocalization of UCNHs.