Extracellular flux analyzer

在过去的十年中，对线粒体生物学的广泛兴趣已经发展，具有通过跨学科方法进行新发现的巨大潜力。作为真核细胞的动力室，线粒体也具有重要的代谢途径，调节钙流变性，并合成Fe-S簇。线粒体功能与细胞器的动态融合和分裂以及调节细胞周期、干细胞性、细胞更新、细胞分化、抗病毒应答和细胞死亡的信号传导途径的细胞内串扰有内在联系。在全身水平上，释放到循环中的丝裂素影响生理功能。对细胞生物能量学和代谢动力学的研究刚刚开始揭示线粒体在控制细胞生命周期、发育和生理学的信号网络中的复杂纠缠。在慕尼黑工业大学，来自生命科学和医学多个学科的研究人员目前正在解决与线粒体生物发生，线粒体自噬，代谢灵活性，活性氧应激，线粒体因子，代谢物，信号传导和细胞钙动力学相关的紧迫研究问题。尽管他们的跨学科分歧，从免疫学，感染生物学，胃肠道病理生理学，癌症生物学，能量平衡，代谢和结构生物学，他们的研究收敛于线粒体在细胞生物能量学控制中的作用，从而触及共同点。这种跨学科的合作在许多高级别期刊（Cell，Nature，Nature Immunology，Nature Communications，Journal of Hepatology等）上发表。这些研究人员需要最先进的技术来进行类器官，完整或透化细胞以及分离的线粒体中的代谢通量测量。需要并行实时记录耗氧率（OCR）和细胞外酸化率（ECAR），以量化提供三磷酸腺苷（ATP）的关键代谢途径的通量。氧消耗率反映线粒体氧化磷酸化活性，而ECAR是糖酵解ATP合成的替代指标。所需的仪器必须允许在最小体积的有限数量的类器官、细胞或线粒体的情况下进行可重复的OCR和ECAR测量，因此需要高灵敏度和精确的温度和湿度控制。评估代谢通量响应于不同激活剂和抑制剂的动态变化是一个基本要求。为了在相同的实验条件下比较不同的处理，必须使用多板系统。

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.