Single-Cell Culture of Bacteria Within Giant Vesicles

Begin with a chambered glass slide coated with a supported membrane composed of phospholipids, including biotinylated phospholipids.

Incubate with neutravidin, which binds to biotin on the bilayer. Wash to remove the excess neutravidin.

Introduce a nutrient medium containing giant vesicles or GVs. Each GV is a spherical, micron-sized lipid bilayer vesicle that encapsulates a single bacterial cell, serving as a model to study individual cell behavior in a confined environment.

Neutravidin binds to biotinylated GV lipids, securing the vesicles to the supported membrane for stable, long-term observation under static conditions.

Seal the chamber to maintain hydration and minimize contamination.

Place the slide on a microscope stage equipped with a heating platform to simulate physiological conditions and observe under a microscope.

Nutrients from the medium diffuse into the GV to support bacterial growth over time.

This setup allows real-time visualization of single-cell bacterial growth within confined vesicle microenvironments.

To immobilize giant vesicles on the supporter bi-layer membrane, introduce 10 microliters of neutravidin in outer aqueous giant vesicle solution to the hole for a 15