Flow cytometric pulse form analysis and sorting for microbiota cytometry

• Applicants: Professor Dr. Hyun-Dong Chang; Toralf Kaiser
• Subject Area: Microbial Ecology and Applied Microbiology ; Cell Biology
• Term: since 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 491069896

Flow cytometry allows the precise and high-throughput fluorescence-based multi-parametric analysis and sorting of single cells and has become the standard single cell technology in biomedical research. However, flow cytometry is optimized for the analysis of eukaryotic cells. In this research project, a new device will be developed that measures the pulse shape of cell-intrinsic scattered light with spatial and temporal resolution for the precise determination of size and shape of small particles or cells. To develop the device, we will 1) develop the optical setup for angle- and time-resolved detection of the scattered light; 2) develop algorithms for data and signal processing; 3) implement a cell sorting unit; and 4) validate the laboratory setup of the device using biological samples. This device will, for the first time, enable quantitative and qualitative single cell analyses and cell sorting based on cell-intrinsic light scatter properties in a high throughput manner in microbiota research. By allowing the label-free and direct sorting of non-culturable bacteria and detection and enrichment of rare bacteria, this device will significantly expand microbiome analyses, which are currently limited mainly to molecular biology methods. In addition, such a device has the potential to significantly expand the application of flow cytometry to other scientific fields, such as marker-free cell analysis, analysis of morphologically heterogeneous cell mixtures, micro- and nano-particles, extra-cellular vesicles, and organoid analysis.

• DFG Programme: New Instrumentation for Research