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Advanced biodetection and bioimaging require fluorescent labels which exhibit many, easily distinguishable colors to identify or study numerous biotargets in a single sample. Although numerous different colors have been demonstrated with lanthanide doped nanoparticles, these colors usually originate from various ratios of overlapping multiple emission bands from activators, which severely limits the number of available labels. As a consequence, different lanthanide doped labels cannot be easily distinguished from each other (e.g. Er3+ from Ho3+) in a quantitative way, when such labels are co-localized during microscopy wide-field imaging. It is therefore reasonable to expand the available choice of spectral signatures and not rely on just different colors. Other ions, such as Tb3+ or Eu3+, can offer new possibilities and unique spectral features in upconversion mode in this respect. For example, despite partial overlap with Er3+ or Ho3+ emission spectra, Tb3+ ions display also unique and easily distinguishable spectral features at 580 nm. Unfortunately, in terms of brightness, Tb3+ emission in upconversion mode is typically too weak to be useful. To improve the Tb3+ upconversion emission intensity, a new approach, i.e. Mn2+ co-doping, has been proposed and verified in this work. A versatile optimization of Tb3+, Yb3+ and Mn2+ ion concentrations has been performed based on luminescence spectra and lifetime studies. The most intense emission was achieved for nanoparticles doped with 10% Mn2+ ions, with over 30 times brighter intensity of Tb3+ ions compared to the emission of nanocrystals without the addition of Mn2+ ions. Additionally, as a proof of the concept, the surface of nanoparticles was coated with proteins and conjugated with folic acid, and such biofunctionalized nanoparticles were subsequently used for bioimaging of HeLa cells.

nanoscale advances

Near-infrared excited luminescence and in vitro imaging of HeLa cells by using Mn2+ enhanced Tb3+ and Yb3+ cooperative upconversion in NaYF4 nanocrystals