4-Laser Spectral flow cytometry analyzer

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 574395861

Recent technological advances in the life-sciences have tremendously propelled our understanding of complex cellular processes. In particular, multi-modal approaches enable the integration of information obtained on the molecular level, for example by next-generation sequencing or proteomics approaches, and on the level of individual cells by various single-cell approaches. The requested spectral flow cytometry analyzer enables rapid extraction of information on fluorescent molecules from single cells, and is thereby particularly well-suited for studies that bridge molecular and cellular scales. The research groups participating in this proposal have already gained significant insights into important fundamental processes at the molecular and cellular scales, most notably in the contexts of cell division, DNA replication and repair, epigenetics, growth factor signalling and cancer development. Compared to conventional devices, the spectral flow cytometry analyzer implements spectral detectors that significantly improve its ability to distinguish different fluorophores or different fluorophore states in single cells using spectral unmixing. This major technological advance enables new possibilities to investigate molecular processes in the cellular context, for example by enabling sensitive fluorescence-based detection of biomolecules at high background autofluorescence levels that are present in several cell types, including the important budding yeast model cell system. Furthermore, spectral flow cytometry enables the parallel detection of a large number of fluorophores that differ only slightly in their spectral properties. Most notably, this allows the sensitive detection of molecular changes within fluorescent probes that are linked to cellular functions, for example due to Förster resonance energy transfer in activity sensors. Moreover, single cell measurements performed via spectral flow cytometry analysis will be combined with omics technologies, such as next generation sequencing or proteomics, to tackle particularly complex biological processes, such as the DNA damage response or proteostasis of the genome integrity network. Processes that occur at very short timescales, such as growth factor signalling, will be addressed by combining rapid optogenetic or pharmacologic perturbations directly at the sample within the spectral flow cytometry analyzer. Taken together, the unique capabilities of spectral flow cytometry to bridge molecular and cellular scales will enable detailed mechanistic insights into biomolecules in the context of individual cells and will thereby facilitate a more holistic understanding of fundamental cell biological processes.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 4-Laser Spektral-Durchflusszytometer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Technische Universität Dortmund
• Leader: Professor Dr. Boris Pfander