5-laser spectral -analysis flow cytometer

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 471514137

In the last 2-3 years there has been a noticeable shift in applications in flow cytometry towards multiparametric analyzes in Bonn. The 3-laser systems, some of which have been in use for over 10 years and for a long time at a consistently high level (average approx. 1200 hours / year), whereas the use of the two 5-laser devices (LSR Fortessa) has steadily increased. After purchasing the second 5-laser analyzer in the summer of 2018, a similarly high level of utilization as the existing device was achieved almost right from the start. Both devices are now close to their utilization limitalmost 2000 hours of booked time in the last year. The scientists are therefore often forced to carry out their measurements until late at night. The spectral flow cytometer applied for here is intended to relieve the existing infrastructure.In addition, spectrally operating flow cytometers, with identical equipment with excitation light sources, offer advantages over conventional flow cytometersSignificantly expand the range of applications for multiparametric flow cytometry. This includes the use of dyes with closely related emission spectra or the subsequent processing of the entire spectrum through “spectral unmixing” and “autofluorescence extraction” to improve the signal quality and thus the sensitivity. The use of fluorescent proteins in model organisms, the tissue-specific autofluorescence associated therewith and the problems resulting therefrom in a subsequent multi-parametric immunophenotyping are a well-known example of this.Novel applications e.g. in the field of markerless phenotyping and theProcessing of the extensive data sets in bioinformatics are the subject of thiscurrent research.A survey of the users of the Core Facility was able to determine that for a large majority of potential future users, the spectral flow cytometry from therepresents the preferred technological expansion of the infrastructure of the core facility for the reasons mentioned above. The interviewed scientists cover the most diverse areas of medical and scientific research at the university. The main ones to be mentioned here are: immunology, neurosciences, cancer research, infectious diseases and cardiology. Many of the research projects summarized in this application would benefit directly from the advantages of spectral analysis mentioned or can in fact only be carried out with such a device. Detailed information on this can be found under point 3 "Use in research".

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 5-Laser Spektrales Analyse-Durchflusszytometer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Rheinische Friedrich-Wilhelms-Universität Bonn
• Leader: Professor Dr. Christian Kurts