Linking structural information of protein interactions, multi-molecular complexes and macromolecular clusters to cellular information is key to understanding how these molecular interactions translate into cellular responses. The Molecular Imaging theme aims to develop and use 3D single molecule and super-resolution microscopy to bridge this gap between structural biology and cellular function. To achieve this aim, we will focus on developing:
- microscope hardware improvements such as novel laser systems, fast cameras, novel optics and sensitive detector systems and corresponding electronics; and
- novel algorithms for image correlation spectroscopy, pair-correlation analysis and variance analysis that can be applied to a range of different imaging techniques.
Fluctuation-based imaging of nuclear Rac1 activation by protein oligomerisation
Centre collaborator Elizabeth Hinde and Deputy Director Kat Gaus were part of a team of researchers who developed a new imaging technique that can quantify the interactions between protein molecules. The technique involves acquiring a time series of images in a particular region of interest of the cell and measuring the fluorescence intensity and lifetime at each pixel in the image. Using this technique, they showed that upon DNA damage, monomeric and active Rac1 in the nucleus was segregated from dimeric and inactive Rac1 in the cytoplasm.
The work was published in Scientific Reports, a Nature group journal, and is a key technique that provides scientists the context in which molecular interactions occur and the ability to follow them in time.
Fluctuation-based imaging of nuclear Rac1 activation by protein oligomerisation. Scientific Reports, Feb 2014
Authors: E Hinde, K Yokomori, K Gaus, KM Hahn, E Gratton
John Davey, Enrico Gratton, Markus Sauer, Christian Eggeling