Flow cytometric analysis has proven valuable for investigating pure cultures and monitoring microbial community dynamics. We present three comprehensive workflows, from sampling to data analysis, for pure cultures and complex communities in clear medium as well as in challenging matrices, respectively.
Total time
~4–6 hours per sample set (including staining, calibration, and analysis)
Steps
1
Dry biogas and stabilize activated sludge communities
Prepare biogas community samples through drying procedures and stabilize activated sludge community (ASC) cells using appropriate preservation methods prior to analysis.
▶ 00:50
2
Stain activated sludge community samples
Apply fluorescent staining protocols to ASC samples to enable flow cytometric detection and discrimination of microbial populations.
▶ 02:29
3
Calibrate beads and analyze ASC samples
Perform bead calibration for flow cytometry and conduct sample analysis on stained ASC to establish baseline cytometric parameters.
▶ 03:46
4
Generate cytometric barcode profiles
Create cytometric barcodes from flow cytometry data to characterize and track microbial community composition and dynamics.
▶ 05:29
5
Interpret microbial evolution and community changes
Analyze representative cytometric datasets to assess temporal microbial community dynamics and evolutionary patterns.
▶ 07:21