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

该项目的两个关键目标是制备抗体功能化的荧光纳米颗粒，这种纳米颗粒具有优异的胶体稳定性和B)可以在一系列复杂的溶液中以高特异性结合到目标分析物上。A)将使用定制的表征方法筛选一系列纳米颗粒稳定方法，包括光散射技术(动态和静态)和荧光相关光谱(FCS)。胶体表征将通过史蒂文斯小组广泛的纳米颗粒经验得到便利(Creamer，Stevens等人，NAT。B)分析物结合将通过石英晶体微天平(QCM)进行筛选。古尔丁博士的团队经常将纳米颗粒固定在QCM上，以研究纳米颗粒与所选分析物的相互作用，以及与复杂溶液中其他蛋白质的非特异性相互作用(Yang，Guldin等人，NanScale，2019&Suthar，Guldin等人，分析化学，2020)。这项技术将被用来指导和通知生物连接的优化。这些目标的实现将有助于使用荧光团进行荧光免疫染色，这些荧光团比传统染料(如AlexaFluor488)更明亮、更稳定，从而为长期跟踪复杂环境中的生物过程打开了可能性。

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.