Fluorogenic Probes for Real-Time Tracking of Bacterial Cell Wall Dynamics with Nanoscopy

Area of Science:

• Microbiology
• Cell Biology
• Biochemistry

Background:

• The bacterial cell wall, primarily peptidoglycan (PG), is vital for cell structure and survival.
• Monitoring PG synthesis and dynamics in live bacteria with high spatial resolution is challenging.

Purpose of the Study:

• To develop novel fluorogenic probes for real-time, super-resolution imaging of bacterial PG synthesis.
• To enable quantitative analysis of PG dynamics and cell division processes in live bacteria.

Main Methods:

• Design and synthesis of rhodamine-based fluorogenic probes with optimized self-aggregation.
• Application of probes for wash-free labeling and imaging across various bacterial species.
• Utilizing 2D and 3D STED microscopy for high-resolution visualization of PG synthesis and cell division machinery.

Main Results:

• Probes exhibit reduced background fluorescence and enhanced fluorogenicity upon labeling.
• Successful real-time and super-resolution imaging of PG synthesis patterns in live bacteria.
• Quantification of septal PG (sPG) synthesis rates and simultaneous imaging of cell division components.

Conclusions:

• The developed probes provide a powerful toolkit for studying bacterial cell wall architecture and dynamics.
• This approach facilitates new research avenues into PG-related cellular processes and bacterial growth.