Evaluating phage lytic activity: from plaque assays to single-cell technologies

Area of Science:

• Microbiology
• Molecular Biology
• Virology

Background:

• Bacteriophages are ubiquitous and vital to microbial ecosystems.
• Numerous techniques exist to study phage-bacteria interactions, evolving over time.
• Phage biology, host interactions, and therapeutic applications are rapidly advancing fields.

Purpose of the Study:

• To provide a comprehensive overview of methods for studying bacteriophage-bacteria interactions.
• To guide researchers in selecting appropriate techniques for phage biology research.
• To celebrate the 110th anniversary of phage discovery.

Main Methods:

• Classical methods: Plaque assays on solid media for phage quantification and host-range determination.
• Liquid culture methods: Optical density, qPCR, metabolic, and cell damage assays for infection dynamics.
• Single-cell techniques: Microscopy, microfluidics, next-generation sequencing, and Hi-C for dissecting infection heterogeneity.

Main Results:

• Diverse techniques, from bulk to single-cell, enable detailed study of phage-bacteria interactions.
• Integration of methods enhances understanding of phage infection mechanisms and bacterial immunity.
• Advancements facilitate the development and application of phage therapy.

Conclusions:

• A wide array of techniques is available for studying bacteriophages and their interactions with bacteria.
• Selecting the right method is key for progress in phage biology and phage therapy.
• Continued methodological innovation drives discoveries in phage-host dynamics and therapeutic potential.