Tracking the delivery of nanoparticles into cells

Supervisor Organisation PhD Awarding Entity: phd location
University College Dublin
University College Dublin
University College Dublin, Ireland

Research Focus

Nanoparticles are increasingly being used as therapeutic delivery vehicles, with more than 40 nanomedicines approved for clinical use. A critical aspect of their further development is gaining a greater understanding of their mechanisms of cellular internalisation and their subcellular distribution, as this knowledge will aid their efficacy for drug delivery. Currently, combinations of transmission electron microscopy (TEM) and fluorescence microscopy are used to assess their localisation in cells, but these methods are limited by their long preparation times, and the inconsistent imaging ability and fluorescent labelling of certain nanomaterial types. Soft X-ray tomography (SXT) will be evaluated as an alternative or complementary method to systematically assess and quantify nanoparticle uptake and distribution in cells, using nanomaterials derived from polymers, silica, metals, carbon and PLGA.

Methodology

This project will utilise a wide range of animal cell culture systems, including monolayer (2D) cultures of cells representing various tissues and organs, as well as small spheroid (3D) models representing mini-tumours. The student will develop expertise in animal cell culture as well as how to manipulate these cells for imaging purposes, including transfection and immunostaining.

The student will also learn critical techniques in characterisation and use of nanoparticles made of various materials. These nanoparticles will be used in cell-based assays, and a wide range of biological imaging techniques (fluorescence light microscopy, confocal microscopy, high-content screening microscopy, electron microscopy, FIB-SEM microscopy) employed, and their performance compared to that of SXT microscopy. The student will also develop expertise in quantitative analysis across these imaging modalities such that they are able to precisely define the mechanics of nanoparticle uptake and distribution in the cellular models, as well as provide a systematic evaluation of the strengths and weaknesses of the various imaging techniques.

Aim 1

Quantitative analysis of nanoparticle uptake into cellular models (2D and 3D)

Aim 2

Nanoparticle types include polymers, lipids, PLGA, carbon, novel materials

Aim 3

Assessment of NP distribution in cells is at subcellular scale

Aim 4

Explore capacity of SXT imaging to evaluate NP distribution in cells

Pictures Attached

Image description: The images show a selection of cell types and models used for advanced fluorescence imaging applications to study nanoparticle uptake. a) various spheroid models stained with fluorescent markers; b) characterisation of polymeric nanoparticle uptake into a spheroid at subcellular resolution and showing the distribution of the nanoparticles (NPs) with respect to the lysosomes.

PhD Researcher

Name: Madeleen Brink

University: University College Dublin

Supervisor’s Name: Prof Jeremy Simpson

Madeleen Brink completed her Bachelor’s and Honour’s degrees at the University of the Free State, South Africa. Following this, she finished her Master’s in Health Science at the Laser Research Centre, University of Johannesburg, South Africa. She is set to pursue her PhD under the supervision of Prof Jeremy Simpson with the CLEXM Consortium, a Marie Sklodowska-Curie Doctoral Networks Action (MSCA-DN) funded by the European Union under Horizon Europe. Her project aims to track the delivery of nanoparticles into cells by applying correlative microscopy. Soft X-ray tomography (SXT) will be evaluated as an alternative or complementary correlative microscopy method to systematically assess and quantify nanoparticle uptake and distribution in cells.

Her research interests lie in correlative microscopy, exploring its applications across diverse imaging modalities to gain comprehensive insights into biological systems. Her research focuses on refining sample preparation techniques, leveraging advanced imaging analysis, and processing methodologies. Overall, her work is driven by a dedication to advancing the capabilities and understanding of imaging technologies in the context of cell biology.

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