Spectral flow cytometer

• Subject Area: Medicine
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 499549016

This request is for the addition of a spectral flow cytometer to the Core Facility for Flow Cytometry at the Campus Innenstadt (LMU Klinikum) to meet the needs for cutting-edge research in the coming years. The Core Facility is used by research groups focusing on immunology, tumor immunology, autoimmunity and vaccinology. With conventional flow cytometers, technically only a limited number of colors can be analyzed simultaneously. In the Core Facility three flow cytometers are currently in use. The most modern instrument in the facility allows the discrimination of 16 parameters. This is not sufficient for the measurement of many newly defined cell types or an exploratory analysis of immune cells. Spectral analysis clearly shifts this limitation. With currently 40 different colors that can be measured simultaneously, a similar order of magnitude is achieved as with the much more expensive and complex mass cytometry. By measuring many parameters simultaneously, even patient samples with limited cell numbers can be defined with high resolution. Spectral analysis offers the following advantages: - the significantly improved sensitivity allows improved resolution of low-expressed surface antigens by photon measurement in the whole spectrum;- similar fluorochromes can also be distinguished by whole spectrum analysis; - fluorescent proteins can be better differentiated, significantly increasing experimental possibilities;- cell population-dependent subtraction of autofluorescence offers a distinct advantage in the analysis of low-expressed markers on cells with strong autofluorescence, e.g. granulocytes;- in addition to the advantages described above, the requested device has an integrated plate reader so that a large number of samples (e.g., in a 96-well plate) can be analyzed automatically. This increases reproducibility and accelerates the throughput rate.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Spektral-Durchflusszytometer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Ludwig-Maximilians-Universität München
• Leader: Professor Dr. Stefan Endres