Feelling Fundamental Principles of Bacterial Cell Physiology using Long-Term Time-Lapse Atomic Force Microscopy

Exposure of bacteria to cidal stresses typically select for the emergence of stress-tolerant cells refractory to killing. Stress tolerance has historically been attributed to the regulation of discrete molecular mechanisms, including though not limited to regulating pro-drug activation or pumps abrogating antibiotic accumulation. However, fractions of mycobacterial mutants lacking these molecular mechanisms still maintain the capacity to broadly tolerate stresses. We have sought to understand the nature of stress tolerance through a largely overlooked axis of mycobacterial-environmental interactions, namely microbial biomechanics. We developed Long-Term Time-Lapse Atomic Force Microscopy (LTTL-AFM) to dynamically characterize nanoscale surface mechanical properties that are otherwise unobservable using other established advanced imaging modalities. LTTL-AFM has allowed us to revisit and redefine fundamental biophysical principles underlying critical bacterial cell processes targeted by a variety of cidal stresses and for which no molecular mechanisms have previously been described. I aim to highlighting the disruptive power of LTTL-AFM to revisit dogmas of fundamental cell processes like cell growth, division, and death. Our studies aim to uncover new molecular paradigms for how mycobacteria physically adapt to stress and provide expanded avenues for the development of novel treatments of microbial infections.