Extracellular flux analyzer

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 508308804

A broad interest in mitochondrial biology has evolved during the past decade with huge potential for new discoveries by cross-disciplinary approaches. Heralded as the powerhouse of eukaryotic cells, mitochondria also harbour essential metabolic pathways, regulate calcium rheostasis, and synthesize Fe-S-clusters. Mitochondrial functions are intrinsically linked to the dynamic fusion and fission of organelles and the intracellular crosstalk of signalling pathways regulating cell-cycle, stemness, cell renewal, cell differentiation, anti-viral responses, and cell death. On the systemic level, mitokines released to the circulation impact physiological functions. Investigations on the dynamics of cellular bioenergetics and metabolism just start to uncover the intricate entanglement of mitochondria in the signalling networks controlling the life cycle of cells, development, and physiology. At the Technical University of Munich, researchers from multiple disciplines working in life sciences and medicine currently address urging research questions related to mitochondrial biogenesis, mitophagy, metabolic flexibility, reactive oxygen species stress, mitokines, metabolites, signalling, and cellular calcium dynamics. Despite their interdisciplinary divergence, ranging from immunology, infection biology, gastrointestinal pathophysiology, cancer biology, energy balance, metabolism, and structural biology, their research converges on the role of mitochondria in the control of cellular bioenergetics and thereby touches common ground. This interdisciplinary collaboration is documented by numerous common publications in high-ranking journals (Cell, Nature, Nature Immunology, Nature Communications, Journal of Hepatology and others).These researchers require state-of-the-art technology to conduct metabolic flux measurements in organoids, intact or permeabilized cells, and in isolated mitochondria. Parallel real-time recordings of oxygen consumption rate (OCR) and extracellular acidification rates (ECAR) are needed to quantify the flux of key metabolic pathways providing adenosine triphosphate (ATP). Oxygen consumption rates reflect mitochondrial oxidative phosphorylation activity whereas ECAR is a surrogate measure for glycolytic ATP synthesis. The required instrumentation must allow reproducible OCR and ECAR measurements with a limited number of organoids, cells or mitochondria in minimal volumes, thus demanding high sensitivity and precise control of temperature and humidity. Assessment of dynamic changes in metabolic flux in response to different activators and inhibitors is an essential requirement. For comparisons of different treatments under identical experimental conditions, a multiplate-system is mandatory.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Extrazellulärer Flux-Analyzer
• Instrumentation Group: 3560 Warburg-Apparaturen, Zellstoffwechsel-Analysengeräte
• Applicant Institution: Technische Universität München (TUM)
• Leader: Professor Dr. Percy Alexander Knolle