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Dr Steven Lee

Portrait of sl591

Dr. Steven F. Lee is a Royal Society University Research Fellow & a Fellow of Sidney Sussex College, Cambridge.

"Building new tools to study biomolecules..."

 

Located in the Chemical Laboratories at the University of Cambridge, TheLeeLab focusses on the development of new biophysical and single-molecule fluorescence methods to answer fundamental biological problems.

Dr. Steven F. Lee

Room G62 - Department of Chemistry
University of Cambridge
Lensfield Road
Cambridge
Cambridgeshire
United Kingdom
CB2 1EW
+441223 331509

 

Research Interests

 

TheLeeLab focuses on many problems related to new biophysical method delveopment, some of which are detailed below:

There is a fundamental limit to how small we can see objects, caused by the diffraction of light itself, this is called Abbe's diffraction limit. This innate 'blurriness' means that optical microscopy cannot resolve anything smaller than ~250 nanometres. Unfortunately the spatial scale that many biological processes occur on is smaller than this limit, therefore it has been difficult to directly probe these events.

Super-resolution microscopy is an advanced, interdisciplinary optical imaging technique currently attracting immense interest as a new and exciting way to break this theoretical diffraction limit of visible light. Using single molecule control, super-resolution microscopy retains the non-invasive advantages of fluorescence imaging but has the ability to resolve biological structures more compatible with the spatial scale that these events actually take place on, typically attaining resolutions of ~15nm or better.

We use novel approaches of super-resolution microscopy in both 2D and 3D, to directly visualise biological processes going on inside living cells at two physical areas: in the eukaryotic plasma membrane and in the nucleus, highlighted by two key areas:

1) The molecular origins of human immunity by studying the spatial distribution and interaction of individual membrane protein complexes on the plasma membrane of live T-cells during triggering.

2) How the molecular composition, stoichiometry and spatial arrangement of histones assemble during DNA replication and repair, all inside the nucleus of fission yeast in three dimensions.

Figure 1. (Upper) The key idea of super-resolution imaging of a structure. (a) It is not possible to resolve the underlying structure in a conventional widefield fluorescence image because the fluorescent labels are in high concentration and the individual emitters overlap. (b) Using controllable fluorophores, it is possible to ‘turn on’ and image a sparse subset of molecules which can then be localised with nanometre precision (grey line is the underlying structure being sampled). Once the first subset of molecules photobleaches, another subset is turned on and localised. This process is repeated and the resulting localisations are summed to give a super-resolution image of the underlying structure.  (Lower) Example of 3D two colour SR imaging of the helical cytoskeletal protein crescentin in the live bacterium caulobacter crescentus (shown in orange/red) in relative context to the cellular membrane (grey) attaining lateral and axial precisions of 15 and 20nm respectively.  Conventional diffraction limited imaging is shown (far left) as a reference, compared with the exploded view of the two component channels (middle images) and then combined (far right). A white light image (inset) and a reference grid (green) made up of 1µm squares is shown for scale. 

These imaging strategies can be applied to a host of other problems where there is need for watching key biological process unfold at high spatial precision at both the membrane and the nucleus. This can only be achieved by attaining a fundamental understanding of the physical and chemical processes behind many aspects of the technology, including: kinetic understanding of fluorophores, theoretical fitting of impulse response functions and detailed knowledge of image reconstruction algorithms .

 

Selected Publications

For an up-to-date publications list, please see our google scholar page

http://scholar.google.co.uk/citations?user=7QbSKnMAAAAJ&hl=en

Publications

A microfluidic platform for trapping, releasing and super-resolution imaging of single cells
Y Zhou, S Basu, KJ Wohlfahrt, SF Lee, D Klenerman, ED Laue, AA Seshia – Sensors and Actuators, B: Chemical (2016) 232, 680
The Nucleosome Remodeling and Deacetylase Complex NuRD Is Built from Preformed Catalytically Active Sub-modules.
W Zhang, A Aubert, JM Gomez de Segura, M Karuppasamy, S Basu, AS Murthy, A Diamante, TA Drury, J Balmer, J Cramard, AA Watson, D Lando, SF Lee, M Palayret, SL Kloet, AH Smits, MJ Deery, M Vermeulen, B Hendrich, D Klenerman, C Schaffitzel, I Berger, ED Laue – Journal of molecular biology (2016)
Initiation of T cell signaling by CD45 segregation at 'close contacts'
VT Chang, RA Fernandes, KA Ganzinger, SF Lee, C Siebold, J McColl, P Jönsson, M Palayret, K Harlos, CH Coles, EY Jones, Y Lui, E Huang, RJ Gilbert, D Klenerman, AR Aricescu, SJ Davis – Nature Immunology (2016) 17, 574
3D Super-Resolution Imaging of Unperturbed Cells
AR Carr, J McCol;, AM Santos, J-E Lee, A Ponjavic, D Klenerman, S Davis, SF Lee – Biophysical Journal (2016) 110, 485a
Improved Photo Physical Properties of mEos3 for Single Molecule Tracking
L-M Needham, S Basu, E Taylor, ED Laue, D Klenerman, D Lando, SF Lee – Biophysical Journal (2016) 110, 485a
Single-Molecule Imaging of Individual Amyloid Protein Aggregates in Human Biofluids
MH Horrocks, SF Lee, S Gandhi, NK Magdalinou, SW Chen, MJ Devine, L Tosatto, M Kjaergaard, JS Beckwith, H Zetterberg, M Iljina, N Cremades, CM Dobson, NW Wood, D Klenerman – ACS Chemical Neuroscience (2016) 7, 399
PSD95 nanoclusters are postsynaptic building blocks in hippocampus circuits
MJ Broadhead, MH Horrocks, F Zhu, L Muresan, R Benavides-Piccione, J DeFelipe, D Fricker, MV Kopanitsa, RR Duncan, D Klenerman, NH Komiyama, SF Lee, SG Grant – Scientific reports (2016) 6, 24626
CalQuo: automated, simultaneous single-cell and population-level quantification of global intracellular Ca2+ responses
M Fritzsche, RA Fernandes, H Colin-York, AM Santos, SF Lee, BC Lagerholm, SJ Davis, C Eggeling – Scientific Reports (2015) 5, 16487
Referenced Single-Molecule Measurements Differentiate between GPCR Oligomerization States
SL Latty, JH Felce, L Weimann, SF Lee, SJ Davis, D Klenerman – Biophys J (2015) 109, 1798
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
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Research Interest Groups

Telephone number

01223 336421 (shared)
01223 331509

Email address

sl591@cam.ac.uk