Single Molecule Imaging: Although fluorescence microscopy has been widely employed as an imaging tool for biological studies, super–resolution fluorescence microscopy allows the 3D mapping of fluorescent proteins in cells with an accuracy far beyond the diffraction limit of visible light. We are currently exploiting super–resolution Photo Activation Localisation Microscopy (PALM) to directly image proteins involved in chromatin assembly and disassembly during DNA replication in live cells at a near molecular resolution. This involves using molecular and cell biological techniques to tag particular proteins with photo–activatable fluorophores in order to study these proteins at the single molecule level. In preliminary studies we have investigated a model epigenetic process – the timing and deposition of the centromere–specific histone H3 variant CENP–A (Cnp1 in S. Pombe) during the cell cycle.
- Quantitative single-molecule microscopy reveals that CENP-A(Cnp1) deposition occurs during G2 in fission yeast, Lando D, Endesfelder U, Berger H, Subramanian L, Dunne PD, McColl J, Klenerman D, Carr AM, Sauer M, Allshire RC, Heilemann M, Laue ED, Open Biol, 2012, 2 (7): 120078
- The S. pombe histone H2A dioxygenase Ofd2 regulates gene expression during hypoxia, Lando D, Balmer J, Laue E D, Kouzarides T, PLoS One, 2012, 7 (1): e29765