Flow cytometer (5 laser system)

• Subject Area: Neurosciences
• Term: Funded in 2019
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 424726578

A key method of immunological, as well as neuroimmunological, research is the analysis of different cell populations via flow cytometry. In immunological research, most multi-parameter analyses of cell populations are performed with high end flow cytometers which allow an easy and detailed analysis of these populations. Flow cytometry allows the researcher to not only label distinct markers via fluorophore-coupled antibodies, but also to analyze different dyes and fluorescent proteins at once. Especially in the new expanding field of neuroimmunology, detailed multi-parameter analysis of different CNS-resident and –associated cell populations become more and more important. Recent studies point to a certain heterogeneity within these immune cell populations in the healthy and diseased CNS. These new findings highlight the complexity, dynamics and plasticity of these CNS immune cell populations, further demonstrating the urgent need of dissecting these cell populations as good as possible in flow cytometry by applying multi-parameter panels for analysis in various settings.A state of the art flow cytometer with five lasers would offer our research a new dimension and flexibility in the choice of fluorophores, dyes and fluorescent proteins. Five laser lines offer a wide spectrum of color choices and allow distributing the markers for the analysis on different laser lines; therefore spill-over effects between different channels can be reduced. Many common markers and fluorescent proteins can be excited by two different laser lines in a five laser flow cytometer and an additional UV-laser would offer new channels for marker analysis. This will lead to a more sensitive detection of the signals in separate channels and a higher resolution between the channels. Spill-over effects or the loss of weak signals can be further diminished by using laser lines with adjustable intensities. A three laser flow cytometer (10 parameters) is so far available in the institute; however this flow cytometer is not sufficient for the planned research projects and the complexity of the research questions. There is an urgent need for a new five laser system to allow us to further pursue the projects and to achieve new advances in our research projects. Therefore, we would like to apply for a new five laser flow cytometer with this proposal.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Durchflussyztometer (5-Lasersystem)
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Albert-Ludwigs-Universität Freiburg
• Leader: Professor Dr. Marco Prinz