Electron microscopy (EM) is an imaging method that provides pictures of specimens at high spatial resolution. EM however has limitations as different molecules cannot be easily distinguished from each other in black and white micrographs. Mass spectrometry (MS) is a structural biology technique that allows the exact chemical identity of molecules in a sample to be determined. The goal of this project is to combine EM and MS imaging to extend the capability of these two methods to study how different biomolecules are arranged within complex multicellular communities of bacteria called biofilms.

Biofilms constitute the majority of bacterial biomass on earth, important in diseases involving antibiotic resistant bacterial infections, and also in health in host-associated microbiomes. Biofilms represent a poorly understood state of bacterial existence on this planet, which needs urgent attention and innovative research to investigate how cells and molecules are arranged within biofilms in three-dimensions.

This proposed project represents basic research, including significant method development. This project is ideal for a technically oriented student interested in microscopy, with an indomitable spirit of exploring uncharted waters. More details will be provided during the interviews.

There is flexibility in biological direction of the project depending on the interests and expertise of the ideal student. There will be opportunities within the project to learn focused ion beam milling of biological specimens, electron tomography (cryo-ET) and advanced data analysis techniques including subtomogram averaging structure determination from cells. We welcome applications from students of all scientific backgrounds.

References

Melia, C., Bolla, J.R., Lanwermeyer, S.K., Mihaylov, D., Hoffmann, P.C., Huo, J., Wozny, M.R., Elfari, L.M., Böhning, J., Owens, R.J., Robinson, C.V., O’Toole, G.A., Bharat, T.A.M. (2021)
Architecture of cell-cell junctions in situ reveals a mechanism for bacterial biofilm inhibition.
Proc Natl Acad Sci U S A,

Chorev, D.S., Tang, H., Rouse, S.L., Bolla, J.R., von Kügelgen, A., Baker, L.A., Wu, D. Gault, J., Bharat, T.A.M., Matthews, S., Robinson, C.V. (2020)
The use of sonicated lipid vesicles for mass spectrometry of membrane protein complexes
Nat Protocols, 1st April

von Kügelgen, A., Tang., H., Hardy, G.G., Kureisaite-Ciziene, D., Brun, Y.V., Stansfeld, P.J., Robinson, C.V., and Bharat, T.A.M. (2020)
In Situ Structure of an Intact Lipopolysaccharide-Bound Bacterial Surface Layer
Cell 180(2): 348-358

Tarafder, A.K., et al., von Kügelgen, A., Mellul, A., Schulze, U., Aarts, D. and Bharat, T.A.M (2020)
Phage liquid crystalline droplets form occlusive sheaths that encapsulate and protect infectious rod-shaped bacteria
Proc Natl Acad Sci U S A,