Flagellar rotational features of an optically confined bacterium at high frequency and temporal resolution reveal the microorganism’s response to changes in the fluid environment

Abstract

Rotations of the flagella control the movement of a peritrichous (multiflagellar) bacterium in fluids, the run and tumble events being caused through modulations in the flagella’s collective rotation speed and pattern. Observing such modulations is a challenge in free swimming bacteria. In this work, we present a setup to measure the collective flagellar rotational features of an optically confined Bacillus subtilis bacterium. We adopt a Continuous Wavelet Technique (CWT) while monitoring the rotational patterns in frequency and time, thus achieving optimal resolution in both the domains. This enables in marking the events wherein subtle changes in the flagellar rotational pattern occur. These studies unravel a fact, hitherto unknown, that variations in swimming speed that are seen in pure run sequences are also caused by modulations in the rotating flagella. Further, we have monitored the flagellar rotation for durations over a minute and observe a gradual slowing down of the rotation before ceasing completely due to the trapping laser induced photodamage. We have observed a significant alteration in the rate of rotational fall off in real time with changes in pH or the nutrient concentration in the fluid. This work serves to demonstrate the advantage of optical confinement of a bacterium in its pristine form for carrying out such studies and can serve as a marker for work that assesses membrane photodamage in active matter. Details on the role of flagella in propulsion and on other factors influencing the rotations, can be of significance in the design of artificial microswimmers.