Time-Resolved Imaging of Bacterial Surfaces Using Atomic Force Microscopy

Area of Science:

• Microbiology
• Biophysics
• Microscopy Techniques

Background:

• Studying bacterial cell physiology requires methods that capture dynamic processes.
• Atomic force microscopy (AFM) provides high-resolution surface imaging.
• Optical fluorescence microscopy (OFM) visualizes molecular events within cells.

Purpose of the Study:

• To detail the materials and methods for time-resolved AFM experiments on bacteria.
• To describe the integration of AFM with OFM for correlative studies.
• To enable the investigation of bacterial physiology across diverse timescales.

Main Methods:

• Utilizing time-resolved atomic force microscopy (AFM) for dynamic bacterial imaging.
• Implementing time-lapse AFM to observe physiological changes over extended periods.
• Performing dual AFM/OFM experiments for correlative analysis of bacterial phenotypes and molecular mechanisms.

Main Results:

• Established protocols for conducting time-resolved AFM on bacterial samples.
• Demonstrated the feasibility of combining AFM and OFM for correlative microscopy.
• Provided a framework for linking physical bacterial phenotypes to underlying molecular processes.

Conclusions:

• Time-resolved AFM is a powerful tool for studying bacterial cell physiology dynamically.
• Correlative AFM/OFM approaches enhance understanding of bacterial phenotypes and molecular mechanisms.
• These methods facilitate comprehensive investigations into bacterial behavior on relevant timescales.