FLUOMECBAC Advanced fluorescence microscopy to understand and improve mechano-bactericidal nanomaterials

Funding :

PID2021-122231NB-I00 PROYECTOS DE GENERACIÓN DE CONOCIMIENTO

Duration:

Mechano-bactericidal nanomaterials, such as low-dimensional nanoparticles in suspension (nanodarts) or high-aspect-ratio nanofabricated topographies, rely on their mechanical or physical interactions with bacteria and are promising antimicrobial strategies that overcome bacterial resistance to classical antibiotics. However, the real effect of mechanical versus chemical action on their activity is under debate. Indeed, fundamental aspects of the physical interaction between bacteria and nanostructures, and the underlying killing mechanisms, are poorly understood. This is in part due to the challenges associated to the characterization of the interface between bacteria and nanostructures in a biologically-relevant environment. In this project, we aim at developing an advanced fluorescence microscopy toolbox to quantitatively study the mechanical effect of nanodarts and nanofabricated topographies on bacterial physiology and viability in real-time and at the single-cell level. We will particularly focus on the range of weak mechanical interactions that lead to bacterial death by fatigue. Three work packages will explore different microscopy methods with increasing level of complexity (single-cell fluorescence imaging in a microfluidic system, fluorescence lifetime imaging and simultaneous force-clamp spectroscopy and fluorescence microscopy), including fluorescence labelling strategies tailored to investigate different aspects of bacterial response. Comparing the physiological response to different nanomaterial geometries and mechanical properties will reveal the purely mechanical component of their bactericidal properties, and will provide design principles for a new generation of mechano-bactericidal materials.