Real-Time Digital Bright Field Technology for Rapid Antibiotic Susceptibility Testing

Area of Science:

• Microbiology
• Biotechnology
• Medical Imaging

Background:

• Bacterial identification and growth rate estimation are crucial for understanding antimicrobial agent efficacy.
• Current methods for antibiotic susceptibility testing (AST) can be time-consuming.
• Automated imaging offers potential for faster and higher-throughput analysis.

Purpose of the Study:

• To describe an automated digital, time-lapse, bright field imaging system for rapid and high-throughput antibiotic susceptibility testing (AST).
• To evaluate the system's capability for analyzing bacterial growth and inhibition in the presence of antimicrobial agents.

Main Methods:

• Utilized an automated digital time-lapse bright field imaging system (oCelloScope) capable of analyzing up to 96 bacteria-antibiotic combinations simultaneously.
• Employed a tilted optical axis (6.25°) to acquire image Z-stacks from microwells, capturing in-focus and out-of-focus bacteria.
• Generated time-lapse videos and analyzed images using dedicated software (UniExplorer) for quantification of morphological parameters, growth, and inhibition.

Main Results:

• The oCelloScope system facilitates rapid and high-throughput antibiotic susceptibility testing.
• The imaging approach allows for real-time monitoring and quantification of bacterial growth and response to antimicrobial agents.
• Image analysis enables the assessment of morphological changes and inhibition over time.

Conclusions:

• Automated optical scanning and image-based analysis provide a robust method for bacterial identification and growth rate modulation studies.
• The described imaging system offers a faster and higher-throughput alternative for antibiotic susceptibility testing.
• This technology has the potential to significantly improve the evaluation of antimicrobial agents.