Groups: Klenerman group website
Telephone: 01223 336481
E-mail: dk10012@cam.ac.uk
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.
Selected Publications
The extracellular chaperone clusterin sequesters oligomeric forms of the amyloid-beta(1-40) peptide. Nature structural & molecular biology 19, 79-83 (2012).
Nanoscale live-cell imaging using hopping probe ion conductance microscopy. Nature Methods 6, 279 - 281 (2009)
Direct characterization of amyloidogenic oligomers by single-molecule fluorescence. Proc. Natl. Acad. Sci. USA 105, 14424-14429 (2008).
Accurate whole human genome sequencing using reversible terminator chemistry. Nature 456, 53-59 (2008).
Single-molecule analysis of human telomerase monomer Nature Chemical Biology 4, 287 - 289 (2008)
Single-molecule level analysis of the subunit composition of the T cell receptor on live T cells Proc. Natl. Acad. Sci. USA 104, 17662-17667 (2007).