Correlative extinction and single fluorophore bleaching microscopy for ligand quantification on gold nanoparticles: data

We report an optical microscopy method to quantitatively evaluate the conjugation on a single particle basis in samples consisting of gold NPs (GNPs) decorated with human holo-transferrin fluorescently labelled with Alexa647 (Tf). We employed widefield fluorescence and extinction microscopy on NP-ligand constructs sparsely deposited onto a glass surface, alongside a correlative analysis which spatially co-localises diffraction-limited sources of fluorescence with the optical extinction by individual GNPs. A photobleaching step analysis of the fluorescence emission was employed to estimate the number of fluorophores contributing to the detected emission rate. The method quantifies the number of fluorescent biomolecules attached per GNP, the numbers of unconjugated GNPs and unbound Tf present within the mixed population, and the size and intraparticle clustering propensity of conjugated GNPs. Data represents single particle analysis taken by fluorescence photobleaching experiments and optical extinction methods presented as tables and charts, as well as electron microscopy images of gold nanoparticles.