研究领域

Physical/Biological

We are physical scientists interested in developing and applying a range of new biophysical methods, based on laser fluorescence spectroscopy and scanning probe microscopy, to important problems in biology, which have not been addressed to date due to the lack of suitable tools. While our experiments range from studies of individual biomolecules to living cells, no previous biological background is required for our research. Single molecule fluorescence In contrast to conventional experiments, which measure ensemble averaged behaviour, single-molecule measurements are able to probe individual molecules to measure variation in their properties and follow their dynamics individually. By studying molecules one at time, specific complexes in a mixture can be identified and analysed without the need for any separation. With our collaborators we are exploiting single molecule fluorescence spectroscopy to probe the intramolecular dynamics, conformations and function of range of biologically important molecules and processes including the T-cell receptor on live cells and protein folding. In collaboration with Professor Dobson we are also probing the early stages of the oligomerisation of proteins involved in many neurodegenerative diseases. Live cell Imaging and Bionanotechnology In collaboration with Professor Korchev at Imperial College we have developed a method based on a scanning nanopipette that allows robust, high resolution, non- contact imaging of living cells, down to the level of individual protein complexes. It can also be used to probe function by performing nanoscale assays such as locally deliver controlled amounts of reagents or performing single ion channel recording. The figure shows the University of Cambridge crest written in fluorescent DNA. We are using this to watch the details of biological process taking place on the surface of living cells, including viral entry and probe the structure of the cell membrane.

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Lipid peroxidation is essential for α-synuclein-induced cell death PR Angelova, MH Horrocks, D Klenerman, S Gandhi, AY Abramov, MS Shchepinov – J Neurochem (2015) n/a (DOI: 10.1111/jnc.13024) Single-molecule FRET reveals hidden complexity in a protein energy landscape. M Tsytlonok, SM Ibrahim, PJ Rowling, W Xu, MJ Ruedas-Rama, A Orte, D Klenerman, LS Itzhaki – Structure (London, England : 1993) (2015) 23, 190 (DOI: 10.1016/j.str.2014.10.023) pyFRET: A Python Library for Single Molecule Fluorescence Data Analysis RR Murphy, SE Jackson, D Klenerman – (2014) Single-Molecule Imaging Reveals that Small Amyloid-β 1-42 Oligomers Interact with the Cellular Prion Protein (PrP C ) KA Ganzinger, P Narayan, SS Qamar, L Weimann, RT Ranasinghe, A Aguzzi, CM Dobson, J McColl, P St George-Hyslop, D Klenerman – Chembiochem (2014) 15, 2515 (DOI: 10.1002/cbic.201402377) Bayesian inference of accurate population sizes and FRET efficiencies from single diffusing biomolecules. RR Murphy, G Danezis, MH Horrocks, SE Jackson, D Klenerman – Anal Chem (2014) 86, 8603 (DOI: 10.1021/ac501188r) Quantification of DNA-associated proteins inside eukaryotic cells using single-molecule localization microscopy TJ Etheridge, RL Boulineau, A Herbert, AT Watson, Y Daigaku, J Tucker, S George, P Jönsson, M Palayret, D Lando, E Laue, MA Osborne, D Klenerman, SF Lee, AM Carr – Nucleic acids research (2014) 42, e146 (DOI: 10.1093/nar/gku726) Rare individual amyloid-β oligomers act on astrocytes to initiate neuronal damage. P Narayan, KM Holmström, DH Kim, DJ Whitcomb, MR Wilson, P St George-Hyslop, NW Wood, CM Dobson, K Cho, AY Abramov, D Klenerman – Biochemistry (2014) 53, 2442 (DOI: 10.1021/bi401606f) Imaging single nanoparticle interactions with human lung cells using fast ion conductance microscopy. P Novak, A Shevchuk, P Ruenraroengsak, M Miragoli, AJ Thorley, D Klenerman, MJ Lab, TD Tetley, J Gorelik, YE Korchev – Nano Lett (2014) 14, 1202 (DOI: 10.1021/nl404068p) Contact-free scanning and imaging with the scanning ion conductance microscope S Del Linz, E Willman, M Caldwell, D Klenerman, A Fernández, G Moss – Anal Chem (2014) 86, 2353 (DOI: 10.1021/ac402748j) The changing point-spread function: single-molecule-based super-resolution imaging. MH Horrocks, M Palayret, D Klenerman, SF Lee – Histochemistry and Cell Biology (2014) 141, 1 (DOI: 10.1007/s00418-014-1186-1) A Single-Molecule Study of Toll-Like Receptor 4 Structure and Signalling SL Latty, KA Ganzinger, LJ Hopkins, C Bryant, D Klenerman – BIOPHYSICAL JOURNAL (2014) 106, 196A Alpha-Synuclein Modulates [Ca2+]c of Neurons and Astrocytes that Trigger Cell Death PR Stroh, MHR Ludtmann, NW Wood, D Klenerman, S Gandhi, AY Abramov – BIOPHYSICAL JOURNAL (2014) 106, 529A Nanopipet Based Nanoprobes for Single-Cell Analysis YE Korchev, P Actis, S Tokar, P Novak, A Shevchuck, D Klenerman, EV Sviderskaya – BIOPHYSICAL JOURNAL (2014) 106, 798A Single Molecule Fluorescence Studies of Protein Aggregates and their role in Neurodegenerative Disease D Klenerman – BIOPHYSICAL JOURNAL (2014) 106, 6A Single Molecule FRET Characterization of Oligomers from Alpha-Synuclein Early Onset Parkinson's Disease Mutants L Tosatto, MH Horrocks, C Nunilo, T Guilliams, M Dalla Serra, D Klenerman – BIOPHYSICAL JOURNAL (2014) 106, 268A Single-Molecule Characterisation of Alpha-Synuclein Oligomers MH Horrocks, SF Lee, S Gandhi, M Iljina, L Tosatto, CM Dobson, D Klenerman – BIOPHYSICAL JOURNAL (2014) 106, 267A Tailoring Nanoprobes for Single-Cell Surgery P Actis, S Tokar, D Klenerman, Y Korchev – BIOPHYSICAL JOURNAL (2014) 106, 414A Tau-Hsp70 Interaction at the Single-Molecule Level F Kundel, M Kjaergaard, SL Shammas, SE Jackson, D Klenerman – BIOPHYSICAL JOURNAL (2014) 106, 59A Utilising Super-Resolution Palm Imaging in Fission Yeast H Armes, T Etheridge, A Herbert, D Lando, SF Lee, D Klenerman, A Carr – BIOPHYSICAL JOURNAL (2014) 106, 199A Label-Free Measurements of the Diffusivity of Molecules in Lipid Membranes B Johansson, F Höök, D Klenerman, P Jönsson – Chemphyschem (2014) 15, 486 (DOI: 10.1002/cphc.201301136)