Flow cytometer

• Subject Area: Medicine
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 501820155

The proposed flow cytometer is intended for the identification of cellular structures which are characteristic for function and origin of diverse cell populations. In experimental studies, the device can be used to identify and quantify specific cell population. Furthermore, the device allows for detailed investigation on the activation and differentiation status of these cells and their function in the respective context. Exemplarily, single cells from blood, from lysed tissues, but also primary cells and cell lines can be used for this purpose. Cells will be stained with fluorescence-labelled antibodies which are directed against specific molecular structures on the cell surface or within the cell. Qualitative and quantitative binding of these antibodies is then detected by the proposed flow cytometer. The obtained data will be processed using specialized software. The proposed flow cytometer facilitates simultaneous analyses of more than 16 parameters on a cell which is a significant advance compared with the already existing device. In consequence, the proposed device is able to analyse even complex cell phenotypes.The increase in the number of detectable parameters at a time facilitates not only a comprehensive characterisation of the cells of interest. Moreover, this is also accompanied by a significant reduction of samples needed which helps to save time and material (antibodies, reagents and buffers etc.). This is particular advantage regarding the small tissue samples that can usually be obtained from experimental tumor mice. Consequently, the device allows a maximum output of information with a minimum need of experimental animals consistent with the requirements of the 3R-(Reduce, Replace, Refine)-guidelines of animal care in life sciences. With the proposed flow cytometer central tasks in funded projects and research networks will be addressed. This includes subprojects from the DFG Transregio-SFB TR209 ‚Liver cancer‘ such as C5 (Wirth), and C6 (Kühnel), and also projects together with cooperation partners of the DFG FOR 2953. The topics of the working groups contributing to this proposal are the development of antitumor vaccines and tumor-directed immunotherapies (groups of Wirth and Yevsa), the identification of tumor-specific effector cells as a method for early detection and prevention of tumor outgrowth (Woller group), breaking of tumor tolerance by using virotherapy (Kühnel group), and the characterization of innate lymphoid cells in hepatocellular cancer (AG Heinrich). Technically, the proposed flow cytometer will be used in projects to characterise the tumor microenvironment and T cells in the course of experimental tumor immunotherapies in diverse gastrointestinal cancers. It is also planned to use the device to a lesser extent for educational purpose and training, particularly as part of student’s lab internships.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Durchflusszytometer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Medizinische Hochschule Hannover
• Leader: Professor Dr. Florian Kühnel