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Controlling Surface Chemical Heterogeneities of Ultrasmall Fluorescent Core–Shell Silica Nanoparticles as Revealed by High-Performance Liquid Chromatography

Ultrasmall (diameter below 10 nm) fluorescent core–shell silica nanoparticles have garnered increasing attention in recent years as a result of their high brightness and favorable biodistribution properties important for applications including bioimaging and nanomedicine. Here, we present an in-depth study that provides new insights into the physical parameters that govern full covalent fluorescent dye encapsulation within the silica core of poly(ethylene glycol)-coated core–shell silica nanoparticles referred to as Cornell prime dots (C′ dots). We use a combination of high-performance liquid chromatography (HPLC), gel-permeation chromatography, and fluorescence correlation spectroscopy to monitor the result of ammonia concentration in the synthesis of C′ dots from negatively and positively charged versions of near-infrared dyes Cy5 and Cy5.5. HPLC, in particular, allows the distinction between cases of full versus partial dye encapsulation in the silica particle core leading to surface chemical heterogen...