Large-Particle Flow Cytometer

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 544252229

Large-Particle Flow Cytometry (LPFC) brings the high-throughput characteristics of conventional flow cytometry to samples of greater size and complexity, facilitating multiplexed quantification approaches of optical parameters such as fluorescence, optical density, and light scattering, as well as isolation of individual animals from heterogeneous populations. The specifications of the targeted large-particle flow cytometer are optimal for approaches using nematode model organisms, including Caenorhabditis along with other species in the phylum. Nematodes are ideal for flow cytometric approaches because their translucent bodies allow for fluorescence-based analysis in living multicellular organisms without dissection. In addition, a range of genetic manipulation tools are well established in C. elegans and other species, thereby opening many exciting research avenues for diverse experimental strategies. With this proposal, we aim to further strengthen the use of LPFC on the Biomedical Campus at the University of Cologne by developing an advanced set of both improved and customized application strategies that will be of great benefit for our researchers as well as new user groups. The principle of LPFC is based on liquid flow-assisted presentation of samples to a detector unit that can quantify fluorescence intensity as well as optical density or light scattering. In addition, images of individual samples can be captured as they pass through the detector unit, allowing for as yet underappreciated analysis options. With the flexibility to monitor multiple optical parameters, virtually any optical readout can be analyzed in a high-throughput manner allowing for remarkably versatile experimental approaches. Of note is that in recent years, LPFC has significantly expanded and improved experimental approaches and data reproducibility for C. elegans and other nematodes. Thus, LPFC constitutes an integral part of high-throughput research approaches using small model organism requiring state-of-the-art technology. Within the frame of purchasing a LPFC instrument, a permanently employed senior scientist will be engaged and supported by lab technician (i) to facilitate the implementation of a dedicated LPFC user platform, (ii) to guarantee central administration of maintenance, optimized use, and long-term conservation of the user experience, and (iii) to enhance and develop high-throughput applications for nematodes to produce data of outstanding quality.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Durchflusszytometer für große und komplexe Probentypen
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Universität zu Köln
• Leader: Professor Dr. Thorsten Hoppe