Overview

Engineered and incidental nanoparticles (NPs) are increasingly discovered in the environment, leading to concerns that they may be harmful to humans and biota. Understanding the potential of NPs to interact and potentially penetrate cell barriers is critical for a full evaluation of their health and safety. Many techniques currently exist that can assess cellular uptake of NPs, but these are either bulk techniques, giving average concentration of NPs in a mass of cells, or are optical methods relying on slow and cumbersome sample preparation and analysis of individual cells.

Single-cell inductively coupled mass spectrometry (SC-ICP-MS, Figure 1) on the other hand, has the potential to revolutionise quantification of cellular load of NPs, by being a high throughput technique, thus measuring a multitude of individual cells, whilst also allowing an accurate measurement of the load within individual cells, as they “fly” through the nebuliser of the single cell ICP-MS. The method is a further development of single particle ICP-MS, a method that is well established at the University of Birmingham.

Nanoparticles entering cells can be identified and quantified using single-cell ICP-MS. Their fate once internalised (e.g. sub-cellular localisation, dissolution or degradation in lysosomes etc.) can also be determined, and potentially modes of action via changes in levels of key intercellular elements.

Methodology

The project aims to use this novel technique, SC-ICP-MS, which is currently available in only a handful of labs around the world, in order to:

1) Gain better understanding of the effects of NP size, surface charge and functionalization on cell-nanoparticle interactions, including uptake quantity, uptake rate and sub-cellular localisation. Well characterised reference NPs (Au, Ag, TiO2), which can be synthesised “in house” will be used to assess any relationship between NP properties and cellular uptake.

2) Assess the stability of NPs within the cellular environment, by assessing changes in NP size over time, and potentially fluxes of metals from the NPs into the cell the cell. Notably the technique is capable of differentiating between dissolved and particulate metal.

3) Develop the method for assessment of biological impacts of NPs, linked directly to the distribution of NPs in different cellular components.

The project will also support UoB’s activities towards developing a better understanding of the fate and behaviour of NPs within the cellular environment and how this may vary as a function of different types of cells (e.g. single cell versus multicellular organisms, and plant versus animal cells).

Training and Skills

CENTA students are required to complete 45 days training throughout their PhD including a 10 day placement. In the first year, students will be trained as a single cohort on environmental science, research methods and core skills. Throughout the PhD, training will progress from core skills sets to master classes specific to the student's projects and themes. 

The student will gain unique access to a well-established analytical lab (FENAC) and develop versatile analytical skills. Although the project work focuses on single particle and single cell ICP-MS, many other analytical methods will be available to the student, from a range of sophisticated microscopic and spectroscopic techniques. The student could, for example, attain analytical skills and expertise working with UV-Vis, DLS, NTA, TEM, AFM, ICP-OES, CE, TGA-IR-GCMS and their data interpretation.

The project will place the student within a large research group, with ample opportunities for internal and external training. The successful applicant will work alongside a set of chemists, environmental, biological and materials scientists with a range of expertise in their area. Full training will be provided by the supervisory team.

Timeline

Year 1: Induction, literature search and laboratory based training on analytical methods focussing on ICP-MS and single cell ICP-MS. Train on cell culture and cellular characterisation using optical methods (e.g. confocal microscopy).

 

Year 2: Development of methodologies for single-cell analytical work. Train on cellular component separation. Apply for Synchrotron access to complement single cell work.

 

Year 3: Develop mechanistic models of cellular uptake of NMs.  Study of distribution of NPs in different cell components. Investigate intracellular behaviour of different size, composition and functionalization NPs. 

Partners and collaboration (including CASE)

The project will be supervised by the host team at the University of Birmingham, with the active participation of project collaborators based at PerkinElmer, and with whom the Birmingham team have an established partnership within the EU funded project ACEnano. Although the student will be based at Birmingham, they will have the opportunity to work in the laboratory facilities of PerkinElmer in the UK and potentially Canada.

 

Further Details

For further project details, please contact Professor Eva Valsami-Jones (e.valsamijones@bham.ac.uk).

See also the following website: www.acenano-project.eu