Principal Supervisor: Professor Pat Unwin, Chemistry
Co-supervisor: Professor Bruno Frenguelli, School of Life Sciences
PhD project title: Lab-on-a-Tip: A New Concept for the Life Sciences and Application to High Resolution Mapping of Neuronal and Glial Activity
University of Registration: University of Warwick
Living cells function through a range of physicochemical processes that take place at cell membranes, as well as within and outside the cell. These processes are becoming accessible to study in unprecedented detail through the use of intelligent nanoprobes, based on multifunctional nanopipettes, which synchronously map and measure cell topography and other properties . This project aims to develop this new experimental platform – largely developed at Warwick – and use it to advance understanding of neuronal and glial activity at the nanoscale. The project is grounded on exciting recent developments from the two supervisors that has already introduced new methods for simultaneous mapping of 3D cell topography and charge [2-4] and sites of molecular uptake at living cells, highlighting the viability of these innovative methods [1,5-7].
We plan to use nanopipette probes that incorporate multiple sensors and which can be further combined with laser scanning confocal microscopy to map neurones and glial cells, and address fundamental questions regarding the utilisation of energy substrates such as glucose, lactate and pyruvate by these cells. These questions are of huge importance because the energy sources of these cells are not known, nor are the responses of the cells to different energy sources.
Further, nanopipette probes will facilitate highly localised manipulations (with fast time resolution) of extracellular pH, oxygen and calcium ions (among others), allowing the response of discrete neuronal and glial elements, such as postsynaptic dendritic spines, presynaptic boutons, and glial release sites, to be identified at unprecedented spatial resolution. These studies will significantly advance understanding of how these cells respond and adapt to transient perturbations in environment, providing an insight to early markers of critical disease states of the brain.
It is expected that this project will provide answers to fundamental aspects of neuronal and glial signalling, and result in the development of instrumentation and analytical techniques, on a platform which is expected to be of wide applicability in the life sciences. As such, the project offers flexibility for important breakthroughs and will allow the student to follow and most exciting avenues. The project will provide experience and training on significant future techniques, associated modelling, image analysis, and cell culture preparation.
- Chen B, Perry D, Teahan J, McPherson IJ, Edmondson J, Kang M, Valavanis D, Frenguelli BG, and Unwin PR, Artificial Synapse: Spatiotemporal Heterogeneities in Dopamine Electrochemistry at a Carbon Fiber Ultramicroelectrode ACS Meas. Au 2021, 1 6–10
- Page A, Perry D, Unwin PR, Multifunctional Scanning Ion Conductance Microscopy (SICM), Proc. R. Soc. A. 2017, A473(2200): 20160889
- Page A, Perry D, Young P, Mitchell D, Frenguelli BG, Unwin PR. Fast Nanoscale Surface Charge Mapping with Pulsed-Potential Scanning Ion Conductance Microscopy. Anal Chem. 2016, 88(22):10854-59.
- Perry D, Paulose Nadappuram B, Momotenko D, Voyias PD, Page A, Tripathi G, Frenguelli BG, Unwin PR. Surface Charge Visualization at Viable Living Cells. J Am Chem Soc. 2016, 138(9):3152-60.
- McKelvey K, Kinnear SL, Perry D, Momotenko D, Unwin PR. Surface charge mapping with a nanopipette. J Am Chem Soc. 2014, 136(39):13735-44.
- Actis P, et al. Electrochemical nanoprobes for single-cell analysis. ACS Nano. 2014, 8(1):875-84.
- Takahashi Y, Shevchuk AI, Novak P, Babakinejad B, Macpherson J, Unwin PR, Shiku H, Gorelik J, Klenerman D, Korchev YE, Matsue T. Topographical and electrochemical nanoscale imaging of living cells using voltage-switching mode scanning electrochemical microscopy. Proc Natl Acad Sci U S A. 2012, 109(29):11540-5.
- Takahashi Y, et al. Multifunctional nanoprobes for nanoscale chemical imaging and localized chemical delivery at surfaces and interfaces. Angew Chem Int Ed Engl. 2011, 50(41):9638-42.
BBSRC Strategic Research Priority: Understanding the Rules of Life: Neuroscience and behaviour
Techniques that will be undertaken during the project:
- Cell culture
- Nanopipette-based techniques and functional imaging
- Laser scanning confocal microscopy
- Quantitative biology
- Image analysis and finite element method modelling
- Instrumentation development