Extracellular Flux Analyzer

在过去的十年里，人们对线粒体生物学产生了广泛的兴趣，并通过跨学科的方法为新的发现带来了巨大的潜力。线粒体被誉为真核细胞的动力源，它还具有重要的代谢途径，调节钙流变性，并合成铁-S簇。线粒体功能与细胞器的动态融合和分裂以及调控细胞周期、干细胞、细胞更新、细胞分化、抗病毒反应和细胞死亡的信号通路的细胞内串扰有内在联系。在系统水平上，丝裂原释放到循环中影响生理功能。对细胞生物能量学和新陈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、发育和生理的信号网络中错综复杂的纠缠。在慕尼黑工业大学，来自生命科学和医学领域的多个学科的研究人员目前正在研究与线粒体生物发生、有丝分裂、代谢灵活性、活性氧物种应激、有丝分裂素、代谢物、信号和细胞钙动力学相关的研究问题。尽管他们在免疫学、感染生物学、胃肠道病理生理学、癌症生物学、能量平衡、代谢和结构生物学等学科之间存在分歧，但他们的研究集中在线粒体在控制细胞生物能量学中的作用上，从而触及了共同点。这种跨学科的合作被许多高级别期刊(细胞、自然、自然免疫学、自然通讯、肝病学杂志等)上的常见出版物所记录。这些研究人员需要最先进的技术来测量有机体、完整或通透性细胞以及分离的线粒体中的代谢通量。需要同时实时记录耗氧率(OCR)和胞外酸化率(ECAR)，以量化提供三磷酸腺苷(ATP)的关键代谢途径的流量。耗氧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内对有限数量的有机物、细胞或线粒体进行可重复的OCR和ECAR测量，从而要求高灵敏度和精确的温度和湿度控制。评估不同激活剂和抑制剂对代谢通量的动态变化是一项基本要求。为了在相同的实验条件下比较不同的处理，必须使用多平板系统。

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.