The optimisation of luminescent lanthanide gold nanoparticles and their detection in cells with mass cytometry

Download Statistics

Downloads

Downloads per month over past year

Bradley, Jamie (2023). The optimisation of luminescent lanthanide gold nanoparticles and their detection in cells with mass cytometry. University of Birmingham. Ph.D.

Preview

Bradley2023PhD.pdf
Text - Accepted Version
Available under License All rights reserved.
Download (13MB) | Preview

Abstract

Gold nanoparticles (AuNPs) are useful scaffolds for the attachment of luminescent lanthanide complexes and are suited for medical applications due to their modifiable surfaces, biocompatibility and suitable size. These properties have led to the extensive study of AuNPs for diagnostic and therapeutic purposes but their translation to the clinic has proved difficult. The work in this thesis focusses on the synthesis and further characterisation of lanthanide coated AuNPs, the quantification of their uptake at the single cell level with mass cytometry (CyTOF), and their potential as multimodal cellular labels for the identification and detection of mononuclear phagocyte differentiation. The synthesis and characterisation of LnQuin\(_{2}\)S-AuNPs and LnS AuNPs and how different chain lengths affect their photophysical properties and cellular uptake is shown. Using electron energy loss spectroscopy, the localisation of europium atoms around AuNPs is demonstrated, showing that the distance from the europium centre to the gold surface is dictated by the organic structure of its ligands. The effects of coating AuNPs with fluorinated surfactants and a PEG polymer on the photophysical properties of EuQuin\(_{2}\)S AuNPs was studied and it was found that fluorinated surfactants enhance their cellular uptake. The use of CyTOF was then further explored as a highly sensitive, single cell quantitative technique to study the interactions of EuQuin\(_{2}\)S AuNPs with HeLa cells, monocytes, macrophages and dendritic cells. Through novel methods of instrument calibration, the quantification of nanoparticle uptake with CyTOF was validated, showing values in agreement with conventional methods. The distribution of EuQuin\(_{2}\)S AuNPs within a cell population was studied and it was found that their depletion from within cells over time is dependent on cell type. The ratiometric detection of gold and europium/terbium from EuQuin\(_{2}\)S AuNPs and TbS-AuNPs from within cells was also demonstrated. With CyTOF, EuQuin\(_{2}\)S-AuNPs can be detected within >99% of macrophages and dendritic cells 7 days after differentiation from NP treated monocytes. Alongside their detection in macrophages by two-photon excitation confocal microscopy, this demonstrated EuQuin\(_{2}\)S AuNPs as potential candidates as multimodal cellular labels.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):