We use a multidisciplinary approach encompassing proteomics, genetic screens, cell biology, biochemistry and structural biology. Basically, we use any technique that helps us to identify and investigate how both host and pathogen determinants alter the outcome of bacterial infections. Some of our favourite techniques are listed here.
We really enjoy using microscopy to visualise infected cells. On the left: a Salmonella-infected cell. Bacteria, expressing GFP are found in a lamp1-positive vacuole (red). The nucleus of the host cell is labelled with DAPI (blue).
We are very fortunate to have access to a Zeiss 710 confocal microscope on our floor as well as a new high-throughput live-imaging instrument via the High Throughput Single Cell Analysis – (HTSCA) Facility that is housed in the CMBI
We also love X-ray crystallography. On the right is the structure of the Salmonella effector and virulence factor GtgA in complex with the N-terminal domain of p65 – a host pro-inflammatory transcription factor that becomes cleaved by GtgA.
For structural biology we work in close collaboration with Dr. Katrin Rittinger at the Francis Crick Institute.
In the CMBI we have access to a state-of-the-art high speed BD FACSAria III sorter, enclosed in a Class 2 microbiological safety cabinet. This allows us to sort cells infected with bacteria prior to analysis. We also use flow cytometry to analyse the amount of bacteria / cell and quantify host protein levels on infected and non-infected cells.