Project R-10789

FABLITE – A light sheet microscopy and microfabrication 'ménage-àdeux' for real-time, high throughput and physiological structural biology. (Research)

In biology, structural methods provide detailed insights into the 3D architecture of biomolecules and are key for e.g. rational drug or biosensor design. Yet, structural biology remains ill-performant when applied to molecules that need structural flexibility to function properly, or to molecules such as membrane proteins, that need a complex physiological environment. In this research project we tackle both issues by developing and combining innovative optical microscopy and microfabrication methods. We establish a novel and broadly applicable fluorescence imaging method based on light-sheet illumination (SPIM) that allows quantifying in real time complex structural rearrangements of biomolecules with, among other things, unprecedented ease and throughput. We also fabricate versatile microdevices in which we embed membrane proteins in a near-native context. Combined with sensitive SPIM we can then uniquely carry out physiologically relevant real-time structural investigations of membrane proteins. We validate our developments and provide many novel insights in the workings the bacterial protein LmrP as a model system for transmembrane metabolite transport and for transporters involved in different human diseases and in drug resistance development in bacteria. This project provides a solid basis for superior membrane protein screening assays and is a seed for many innovative developments and applications in the optical microscopy and device engineering fields.

01 October 2020 - 30 September 2024