Daniel Pencross

Doctoral Researcher
Physical Sciences of Imaging in the Biomedical Sciences CDT

Thesis project - "STORM in a Teacup: Quantification of Receptor-Surface Interactions in Single Cells Through the Application of Novel Recognition Techniques in Super Resolution Images"

Supervisors:
Dr Iain Styles, School of Computer Science
Dr Rob Neely, School of Chemistry
Dr Natalie Poulter, Institute of Cardiovascular Sciences
Dr Steve Thomas, Institute of Cardiovascular Sciences

The principle aim of this investigation will be to develop pattern recognition algorithms capable of both identifying and quantifying structure in super resolution images of fluorescently labeled integrins IIb3 & 21 and other platelet receptors and subsequently use this information to develop a probabilistic model of platelet activation. The project will integrate multiple areas of the physical sciences, primarily focussing on adapting superresolution of microscopy techniques in order to develop applications relating to the study of fluorescently labeled integrins and other receptors on the surface of platelets. To affect these adaptations, the project requires development of new labelling techniques, specically Point Accumulation for Imaging in Nanoscale Topography (PAINT), which makes use of varying DNA oligonucleotides attached to a single fluorophore to produce a theoretically unlimited number of distinguishable signals. Utilising this technique will require the addition of a flow cell system to the existing microscope setup, which in turn will require adaptations to automate the data acquisition process. Validation of the techniques outlined previously will be performed through the use of self assembling DNA molecules designed to emulate the structure of multimeric complexes on the nanometer scale. The ability to design these structures to have known oligo binding sites at specific locations will allow for validation of the PAINT technique outlined previously, as well as testing the spatial resolution achievable by the imaging system. In addition, these structures will be used to mimic motile receptor complexes through programed folding, allowing for testing of molecular tracking algorithms in a controlled environment.