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Microscopy-Based Multiwell Assay to Characterize Disturbed Bacterial Morphogenesis Upon Antibiotic Action
Cruz L Matos de Opitz1, Peter Sass2
1Interfaculty Institute of Microbiology and Infection Medicine, Microbial Bioactive Compounds, University of Tübingen, Tübingen, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|November 29, 2022
Summary
New methods are needed to find antimicrobial drugs targeting bacterial cell division. This study presents a high-throughput microscopy approach to identify compounds that cause bacterial filamentation, aiding drug discovery for resistant infections.
Area of Science:
- Microbiology
- Drug Discovery
- Cell Biology
Background:
- The rise of multidrug-resistant bacteria necessitates novel antimicrobial agents targeting unique pathways.
- Bacterial cell division is a promising target for new antibiotics, with inhibitors often causing bacterial filamentation.
Purpose of the Study:
- To develop a semiautomated, microscopy-based, high-throughput method for identifying and characterizing novel antibacterial compounds.
- To enable real-time observation of compound effects on bacterial morphology (e.g., filamentation, lysis) using minimal compound volumes.
Main Methods:
- A semiautomated, microscopy-based workflow in a 96-well format for observing live bacteria.
- Real-time monitoring of bacterial filamentation and cell lysis induced by novel compounds.
- High-throughput screening of small-volume compound libraries for antibacterial activity.
Main Results:
- The described method allows for the rapid, initial characterization of multiple compounds simultaneously.
- It facilitates the identification of compounds targeting bacterial cell division by observing filamentation phenotypes.
- The approach is suitable for compounds available in limited quantities, common in early drug discovery.
Conclusions:
- A robust, high-throughput microscopy assay is established for identifying novel antibacterial compounds targeting bacterial cell division.
- This method aids in the initial characterization of antibacterial modes of action, particularly filamentation.
- The approach supports efficient drug discovery efforts against challenging bacterial infections.
