In situ characterisation of the bio-nano interface of hybrid fluorescent semiconducting nanoparticles for in vitro

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The two key aims of this project are to fabricate antibody-functionalised fluorescent nanoparticles, which have; A) excellent colloidal stability and B) can bind to the target analyte with a high specificity, in a range of complex solutions.A) A range of nanoparticle stabilisation methods will be screened using bespoke characterisation methods, including light scattering techniques (dynamic and static) and fluorescence correlation spectroscopy (FCS). Colloidal characterisation will be facilitated by the extensive nanoparticle experience in the Stevens group (Creamer, Stevens et al., Nat. Comm., 2018 & Howes, Stevens et al., Science, 2013) .B) Analyte binding will be screened via quartz crystal microbalance (QCM). The group of Dr. Guldin routinely immobilises nanoparticles on QCM to study the interaction of the nanoparticles with the analyte of choice and non-specific interactions with other proteins, in complex solutions (Yang, Guldin et al., Nanoscale, 2019 & Suthar, Guldin et al., Analytical Chemistry, 2020). This technique will be used to guide and inform the bioconjugation optimisation. The fulfilment of these aims will facilitate fluorescent immunostaining with fluorophores which are much brighter and more photostable than the conventional dyes (e.g. AlexaFluor488), opening up the possibility for long-term tracking of biological processes in complex environments.