Evaluation of Respiration with a Clark-Type Electrode in Isolated Mitochondria, Intact and Permeabilized Cells, and...

Evaluation of mitochondrial function in aerobic cells is crucial for understanding the conditions that can potentially compromise their physiology. From the fields of Toxicology to Oncology, various approaches involving freshly isolated fractions of mitochondria, permeabilized cells, intact cells, or animal tissues have been employed to investigate metabolism through oxygen consumption.Several techniques are available for measuring oxygen consumption in liquids. These include polarography with oxygen electrodes, which can employ chemical, electrochemical, or optical detection methods, as well as the use of fluorescent or luminescent probes. In this chapter, we will review the concepts previously discussed in earlier editions, focusing on Clark-type electrodes for electrochemical detection. Additionally, we will explore other approaches that involve intact cells and tissue explants with minimal plasmatic membrane alterations. These techniques provide an integrated view of Glycolysis, Krebs Cycle and Oxidative Phosphorylation. Despite being a classical and cost-effective system, this elegant technique continues to amaze us with its versatility and generation of reliable data.