Synthesis, characterization, and biological applications of multifluorescent silica nanoparticles

Multifluorescent silica nanoparticles were synthesized by the Stöber method using conjugates of (3-aminopropyl)triethoxysilane and fluorescent dye-N-hydroxysuccinimide esters. The nanoparticles containing the fluorescent dyes were well dispersed and showed high, stable, and tunable fluorescence intensities. In addition, we prepared multifluorescent silica nanoparticles containing two kinds of fluorescent dyes and used the nanoparticles in biological applications. Flow cytometry analysis showed high and tuned fluorescence and multiple fluorescences from single nanoparticles with diameters of approximately 400 nm. Fluorescence microscopy analysis also showed high and tuned fluorescence, as well as multiple fluorescences from single nanoparticles and from cells labeled with multifluorescent silica nanoparticles. The intracellular distribution of nanoparticles was evaluated by confocal microscopy and electron microscopy. We discuss the advantages and demonstrate the usefulness of our nanoparticles in relation to commercially available fluorescent nanoparticles including quantum dots.