A microfluidic cell-trapping device for single-cell tracking of host-microbe interactions.

Area of Science:

• Microbiology
• Cell Biology
• Biotechnology

Background:

• Cellular individuality significantly influences host-microbe interactions, but studying these dynamics at the single-cell level is difficult.
• Temporal dynamics of single-cell behavior in host-pathogen systems are crucial for understanding infection outcomes.
• Existing methods often rely on population-based measurements, limiting insights into individual cell responses.

Purpose of the Study:

• To develop and validate a microfluidic platform, InfectChip, for high-resolution, time-lapse microscopy of infected, motile cells.
• To visualize the complete infection process from pathogen entry to host or pathogen cell death.
• To investigate the heterogeneity of host-microbe interactions at the single-cell level.

Main Methods:

• Development of the InfectChip microfluidic device for trapping motile infected cells.
• Utilizing high-resolution time-lapse microscopy to observe cellular events over extended periods.
• Co-culturing the host-cell model Dictyostelium discoideum with Klebsiella pneumoniae (extracellular) and Mycobacterium marinum (intracellular).

Main Results:

• The InfectChip platform successfully visualized all stages of infection dynamics in real-time.
• Observed significant heterogeneity in infection outcomes, including abortive infections and differential cell death.
• Demonstrated the platform's utility in studying both extracellular and intracellular pathogens.

Conclusions:

• InfectChip provides a robust and simple method for dissecting host-microbe interactions at the single-cell level.
• The study highlights the variability in infection outcomes, challenging assumptions based on population-level data.
• This approach yields novel insights into infection dynamics that are not accessible through conventional methods.