Thom Sharp's Research Page

Current research

My research involves the design, characterisation and application of new probes for Correlative Light Electron Microscopy (CLEM). Green Fluorescent Protein (GFP) is a fluorescent protein that can be genetically fused to a protein of interest. When the gene is expressed the location of the GFP-tagged protein can be followed in the live cell using fluorescence light microscopy. The primary aim of my PhD is to generate analogous probes for use in the EM. Several approaches are being attempted, including electron-dense proteins, proteins that bind electron-dense probes in vivo, and probes that produce electron-dense nanoparticles.

GFP – The chromophore is shown in green

GFP-The chromophore is shown in green

My PhD will involve a combination of protein design and engineering, molecular cell biology, light microscopy (confocal, widefield and spinning disk), electron microscopy, and chemistry to generate and test novel electron-dense labels for localization in the EM. Techniques used for characterisation of the probes include localisation along self-assembling peptide fibres, within large unilaminar vesicles and along protein-mediated membrane tubules, as well as MALDI mass spectrometry and dynamic light scattering.

So far I have co-authored one paper, which is now awaiting publication:
"Intracellular membrane traffic at high resolution." JRT van Weering, E Brown, TH Sharp, J Mantell, PJ Cullen & P Verkade. Methods in Cell Biology (Manuscript submitted).

Previous work

In my final year project I worked in the Bristol Royal Infirmary assessing the role played by matrix-degrading metalloproteinases in atherosclerosis. This involved western blotting for fragments of N-cadherin after cleavage, combined with apoptosis assays and fluorescence microscopy.
During undergraduate studies I worked for a year at the pharmaceutical company AstraZeneca in their Safety Pharmacology department. Whilst there I performed hippocampal brain-slice electrophysiology to validate and optimise a new system (SliceMaster) as a medium-throughput screen to detect the convulsant effects of drugs. While there my supervisor and I published a paper of our results:
"Pharmacological validation of a semi-automated in vitro hippocampal brain slice assay for assessment of seizure liability." A Easter, TH Sharp, J-P Valentin & CE Pollard. Journal of Pharmacological and Toxicological Methods 56 (2007) 223-233.