Have an Ion on It: The Life-Cycle of Bdellovibrio bacteriovorus Viewed by Helium-Ion Microscopy

Helium-ion microscopy (HIM) has so far rarely been employed to image microbial interactions. Here, the visualization of the life-cycle of the bacterial predator Bdellovibrio bacteriovorus HD100 with Escherichia coli and Pseudomonas putida, respectively, as prey is presented. The predator is brought in contact with prey and samples are taken at selected times. The system is monitored by phase-contrast microscopy and HIM. For HIM imaging, a sample preparation protocol is established that preserves the structure of Bdellovibrio, prey, and bdelloplasts. The micrographs show the attachment of the predator to its prey, the evolution of bdelloplasts, their lysis, and the release of predator progeny. The combination of HIM with two more approaches allows for investigating predator-prey interactions from different angles: First, phase-contrast micrographs provide quantitative information for the numbers of predator, prey, and bdelloplasts. Second, a numerical model solving the retarded differential equations that describe the system's time-evolution is developed and fits the experimentally determined cell numbers. In conclusion, the high resolution, the large depth of focus, and surface sensitivity of HIM hold promise to expand future studies on so far neglected ecological interactions within the microbial food web, in particular in samples with pronounced topography such as bacterial biofilms.