6-Laser Spectral Flow Cytometry

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 553871989

High-dimensional flow cytometry has become a staple of modern cellular and functional immunology and cell biology. Thus, the need for more advanced multi-parametric analysis has steadily increased. However, the use of conventional flow cytometers for multi-parametric analysis is limited by the need for discrete detection of fluorochromes, and labour-intensive and error-prone compensation processes are needed to make such data interpretable. These limitations result in an upper ceiling of around 20-25 parameters. Additionally, detecting fluorochromes and/or fluorescent reporter proteins in complex tissue settings is severely limited due to the above-described technical specifications. Furthermore, human and mouse non-lymphoid tissues, such as the lung and the liver, exert high autofluorescence. Using conventional flow cytometry, autofluorescence is hard to account for and often results in inaccurate or non-interpretable data. Spectral flow cytometry provides solutions to the aforementioned technical limitation and thus rapidly advances multi-parametric analysis of auto-fluorescent tissues in humans and mice. Further, it allows the inclusion of more fluorescent markers with closer detection spectra as the complete spectrum of fluorescence is accounted for in a process called ‘spectral unmixing’. Within the Life & Medical Sciences (LIMES) Institute of the University of Bonn demand for high-dimensional analysis of hard-to-analyse samples, such as human and mouse non-lymphoid tissues, has steadily increased over the last 5 years resulting in a constant high level of booking of the existing Becton Dickinson FACS Symphony A5. For the above-mentioned reason, we opted for a spectral flow cytometer to maximize the benefit of research conducted at the Life & Medical Sciences Institute and the wider Faculty of Natural Sciences & Mathematics of the University of Bonn by integrating the device into the newly established Flow Cytometry Core Facility – Campus Poppelsdorf (FCCF-CP) of the Faculty of Natural Sciences & Mathematics.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Spektrales 6-Laser Durchflusszytometer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Rheinische Friedrich-Wilhelms-Universität Bonn
• Leader: Professorin Dr. Elvira Mass