FISH and chips: marine bacterial communities analyzed by flow cytometry based on microfluidics

Area of Science:

• Microbiology
• Marine Biology
• Environmental Science

Background:

• Past research utilized PCR and microscopy for marine bacterial community analysis.
• Combining flow cytometry with fluorescence in situ hybridization (FISH) for marine bacteria has been challenging.

Purpose of the Study:

• To develop and validate a microfluidics-based flow cytometry method for analyzing natural marine pelagic bacterial communities.
• To assess the feasibility of using the Agilent 2100 bioanalyzer for FISH analysis of marine bacteria.

Main Methods:

• Marine water samples were filtered and hybridized with Cy5-labeled gene probes.
• Bacteria were detached from filters and analyzed using microfluidics-based flow cytometry (Agilent 2100 bioanalyzer).
• Samples were counter-stained with SYTOX, and FISH signals were analyzed.

Main Results:

• Successful differentiation of bacterial populations using EUB338 and NON probes.
• Demonstrated dominance of alpha-proteobacteria, identified using ALF968 and G rB probes.
• Validated the effectiveness of microfluidics-based flow cytometry for marine bacterial community analysis.

Conclusions:

• Microfluidics-based flow cytometry is a viable and promising technique for analyzing natural marine pelagic bacterial communities.
• This method offers a robust approach for high-throughput analysis of microbial diversity in marine environments.
• The study highlights the prevalence of alpha-proteobacteria in the investigated pelagic marine samples.