Flow cytometer with up to five lasers, spectral analyses and auto-sampling

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 504371625

Our studies seek to understand how the spatial organization and assembly dynamics of plasma membrane signaling pathways control T cell activation and functions. The research is based on our findings that plasma membrane proteins are localized in nanometer-sized domains. Our studies revealed that T cell activation is dynamically controlled by the spatial relationships of distinct nanodomains, the assembly of signaling complexes within them and translocation of signaling molecules between them. We study the underlying molecular mechanisms using multidisciplinary strategies that combine single molecule microscopy with biochemical, biophysical, and cell biological approaches. Specifically, we are interested in T cell receptor (TCR) and the inhibitory program cell death protein (PD-1) pathways during T cell activation and anti-tumor responses. Our ultimate goal is to identify novel control principles in T cell signaling pathways and use them to modify T cell responses for future immunotherapies.We request a flow cytometer with spectral analyzer and autosampler, which is essential to our newly established laboratory at the Albert-Ludwigs-Universität Freiburg and for realizing our current and future research goals. The requested flow cytometer is essential for multiple purposes, including: (i) Studying immune cell diversity during immune responses and in the tumor environment.(ii) Analyses of T cell development, differentiation and activity states. (iii) Quality control of primary and genetically modified cells to maximize success rates and data of our studies, i.e. in vivo anti-tumor responses and advanced single molecule microscopy.(iv) Characterization and testing of new probes and ligands for flow cytometry and fluorescence microscopy.(v) Screening of small molecule libraries, e.g. with FRET probes as read-outs. The flow cytometer has the following two key capabilities:(i) Five laser lines (including UV) for detection of up to 64 fluorophores through spectral analyses.(ii) A 96-wellplate sampler for medium-throughput experiments of cellular and tissue samples, as well as small molecule inhibitor libraries.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Durchflusszytometer mit bis zu fünf Lasern, Spektralanalyse und Autosampling
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Albert-Ludwigs-Universität Freiburg
• Leader: Professor Björn Lillemeier, Ph.D.