In this study we show that steady-state fluorescence anisotorpy within PEBBLEs can be used for the optochemical sensing of analytes such as Zn2+, O2, and Ca2+. Steady-state fluorescence anisotropy is a non- time resolved method that measures a combination of rotational and fluorescence lifetimes. This eliminates the need for reference dyes and ratiometic techniques to obtain quantitative results, even when using intensity-based sensor dyes. An advantage to working with PEBBLE nanosensors is that the encapsulated dye is localized in a constant rotational environment. This is in contrast to the use of free dyes, which can be affected by interferents such as protein binding.
Monodisperse, spherical, polyethylene glycol (PEG)-coated silica nanoparticles have been prepared in the size range of 50-350 nm, and their size distribution were characterized by SEM and multi-angle static light scattering experiments. The chemical binding of PEG to the silica nanoparticles was confirmed by IR spectroscopy. The biocompatibility of these PEGylated nanoparticles was also studied by non-specific protein binding tests and in-vivo toxicology studies in live animals. These silica nanoparticles, as a matrix for encapsulation of certain reagents, have been used for the fabrication of intracellular sensors and have potential for applications to in vivo diagnosis, analysis and measurements, due to their small physical size and their biocompatibility, both stemming from the specialized PEG coating.
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