Live-Cell Time-Lapse Imaging of Bdellovibrio bacteriovorus Predation on E. coli

Begin with a Petri dish containing a co-culture of Bdellovibrio bacteriovorus, a predatory bacterium, and E. coli, its prey, beneath an agarose pad.

The agarose pad keeps E. coli immobilized, while B. bacteriovorus remains motile and actively invades prey cells.

The predator expresses a red cytoplasmic marker and a green fluorescent tag fused to a DNA polymerase subunit for visualization.

Secure the dish in a holder and apply immersion oil to the base of the dish and the microscope objective for optimal resolution.

Mount the holder on the microscope and configure it for dual-channel fluorescence detection.

Use brightfield optics to locate the focal plane and select multiple non-overlapping imaging fields.

Set time-lapse parameters to capture both brightfield and fluorescence images sequentially at defined intervals.

This method enables observation of predation beginning with prey attachment and invasion, followed by prey cell transformation, filamentous growth, septation, progeny formation, and ending with prey cell lysis.

For Time-Lapse Florescence microscopy of the co-culture, place the dish in a Petri dish holder such that the dish will not move over the course of the experiment and pla