FlowSoFine - A tool for analyzing microbial pattern changes detected by flow cytometry

Short read 16s rRNA sequencing is the dominating technology to profile microbial communities in different habitats. Its uncontested taxonomic resolution paved the way for major contributions to the field. Sample measurement, i.e. sequencing, is rather slow – in order of days - and destructive. Taking only minutes, flow cytometry can alternatively be used to profile the microbiome of different sources. It is a high-throughput technology widely used in all research fields examining cell properties. To ease the analytical steps necessary to work out the biological relevant signals from raw measurements via automated gating strategies to publication ready figures, we developed “FlowSoFine”, an interactive, free web application. To validate the ability to discriminate between microbial profiles, we subjected the very same human skin samples of feet, forearm and forehead (n=3*54) to flow cytometry and 16s rRNA sequencing. Loading and processing the raw data, running statistical tests and visualization through our graphical user interface took literally five minutes. As confirmed by sequencing, body site was found to be significantly different through flow cytometry. FlowSoFine provides the tools to boost application of flow cytometry for rapid microbiome analysis. Future developments will leverage the sorting capabilities to isolate differences driving bacteria for downstream experiments.