Tracking Bacterial Growth at Single-Cell Resolution in a Microfluidic System

Begin with a microfluidic device mounted on the stage of an inverted fluorescence microscope.

The device consists of a polymer chip featuring horizontal feeding channels. Each channel is lined with a series of narrow trenches sealed at one end and designed to confine bacterial cells in linear chains.

Within the trenches are non-motile bacterial cells that express a fluorescent protein, enabling their visualization.

Each feeding channel is connected to syringe pumps that deliver nutrient-rich medium at a controlled flow rate.

Initiate the flow to allow the medium to diffuse into the trenches and support bacterial growth.

Confined bacterial cells divide within the trench, forming a chain of progeny.

The trench design ensures newer cells at the open end are washed away, enabling long-term tracking of the oldest cells.

Capture images of trenches over time to track the growth and division of individual bacterial cells.

Carefully mount the loaded device onto a stage insert by taping the cover glass on either side of the PDMS to the bottom of the stage insert. Invert the stage insert device assembly so that the PDMS is facing up and place it on a soft surface such as a dust-free wipe.

Adjust the f