4-Laser Cell Sorter

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 459783289

A flow cytometer is an instrument that detects and measures physical and chemical characteristics of a population of cells or particles providing quantifiable data at the single cell/particle level. The proposed instrument includes a high-speed cell sorter, which physically separates cells of interest based on their optical properties. The proposed flow cytometer will assist molecular biology research within the frame of a multi-user concept. Diverse research topics are aimed ranging from synthetic biology in cyanobacteria, yeast, plant and mammalian cells, towards manipulation and monitoring of human cells, e.g. beta cells, hepatocytes, endothelial cells. Synthetic metabolic pathways for sugar polymer production as well as sugar and hormone biosensors will be engineered and quantitatively characterized at the single cell level, which is key in synthetic biology approaches. Novel, cytometer-based methods will be established like the iTRAP method, which develops proteolytic antibody derivatives targeting peptides involved in Alzheimer’s Disease and type II diabetes, respectively. The establishment of an inducible knock out system is aimed for a trypanosomatid and its proteobacterial endosymbiont as well as isolation and characterization of different cell cycle stages. High-performance optogenetic switches will be engineered and optogenetic switches with custom functionality obtained through directed evolution. Further, a relative quantification of transgenetically-implemented biomolecules will be established using multicolor detection in mammalian systems. Flow cytometer based cell sorting will be used to separate e.g. intestinal cell types according to fluorophore expression, which will allow for elucidation of a newly observed circadian rhythm in intestinal stem cells. In summary, the proposed instrument - a flow cytometer including four different lasers/colors and a high speed sorter - will support a plethora of ongoing research projects and initiatives at our department by providing an absolutely essential, user-friendly and very flexible infrastructure.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 4-Laser Zellsortierer
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professorin Dr. Ilka Maria Axmann