Stripping flow cytometry: How many detectors do we need for bacterial identification?

Area of Science:

• Microbiology
• Analytical Chemistry
• Biotechnology

Background:

• Multicolor microbial flow cytometry faces challenges due to instruments designed for biomedical use, often featuring excess detectors.
• The biological relevance of spectral overlap data from additional fluorescence detectors in bacterial identification is unclear.

Purpose of the Study:

• To characterize the utility of additional fluorescence detectors in microbial flow cytometry.
• To develop a data-driven method for selecting an optimal subset of detectors for bacterial population discrimination.

Main Methods:

• Proposed a data-driven detector selection method using a detector elimination strategy.
• Applied the method to experimental data from two different modern cytometer configurations.
• Analyzed the importance of individual detectors for bacterial population identification.

Main Results:

• Demonstrated that detectors can be removed without compromising the resolving power of bacterial identification.
• Identified a subset of detectors that optimally discriminate between bacterial populations.
• Revealed instrument-specific detector importances and unexpected SYBR Green I behavior in the red spectrum.

Conclusions:

• The developed method effectively identifies essential detectors for microbial flow cytometry.
• Results suggest a need for cytometric instruments tailored for microbiology with reduced, optimized detector numbers.
• Findings pave the way for more efficient and specialized microbial flow cytometry applications.