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Photoacoustic Flow Cytometry Using Functionalized Microspheres for Selective Detection of Bacteria
Robert H Edgar1, Anie-Pier Samson2, Tori Kocsis2
1Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Micromachines
|March 29, 2023
Summary
Photoacoustic flow cytometry offers precise detection of rare analytes. This study demonstrates its use for identifying specific bacteria, like Salmonella, in fluids using modified bacteriophages.
Area of Science:
- Biotechnology
- Microbiology
- Analytical Chemistry
Background:
- Photoacoustic flow cytometry (PAFC) is an emerging technique for detecting rare cells in fluid samples.
- Current methods for identifying specific bacterial species in clinical samples can be time-consuming and lack sensitivity.
- Bacteriophages, viruses that infect bacteria, can be engineered for targeted bacterial recognition.
Purpose of the Study:
- To develop a novel PAFC method for the sensitive and specific detection of bacteria in fluid samples.
- To utilize modified bacteriophages conjugated with microspheres for enhanced optical absorption and targeted bacterial identification.
- To reduce variability and prevent bacterial lysis during the detection process.
Main Methods:
- Engineered bacteriophages were modified by attaching dyed latex microspheres to their tail fibers, preserving bacterial recognition capabilities.
- These microsphere-bacteriophage complexes were used to target specific bacterial species, Salmonella enterica and Escherichia coli.
- The detection specificity and sensitivity were evaluated using PAFC by quantifying targeted bacteria in mixed samples.
Main Results:
- The study successfully demonstrated robust and specific detection of targeted bacteria using the developed PAFC method.
- Experiments using Salmonella (LT2 strain) and E. coli (K12 strain) showed distinct detection signals.
- With red microspheres, an average of 109.3±9.0 Salmonella were detected versus 2.0±1.7 E. coli; with blue microspheres, 86.7±13.2 Salmonella were detected versus 0.3±0.6 E. coli.
Conclusions:
- The developed photoacoustic flow cytometry method, utilizing modified bacteriophages, enables selective and sensitive bacterial species detection.
- This approach offers a promising tool for rapid identification of specific bacteria in various fluid samples.
- The findings highlight the potential of bacteriophage-based biosensors for advancing diagnostic capabilities in microbiology.
