Measure glycolysis and respiration in live cells with a CLARIOstar

The number of questions that demand to study cellular energy metabolism is rising: How is mitochondrial dysfunction implicated in diseases such as cancer or diabetes? How is energy production accomplished without blood supply during strokes and infarcts? Does a novel drug induce toxicity by impairing mitochondria?

New assays simplified and accelerated the investigation of cellular metabolic pathways such as glycolysis and oxidative phosphorylation. Phosphorescent probes report in real-time and microplate format on the metabolic parameters oxygen consumption and extracellular acidification. BMG LABTECH supports these assays by providing high sensitivity detection devices with unique features for metabolic research. The CLARIOstar microplate reader with its Atmospheric Control Unit not only allows to measure cells at physiologic, hypoxic and hyperoxic conditions, but mimics ischemia/reperfusion processes by de- and re-oxygenating the incubation chamber.

The metabolic condition of a cell sheds light on its state of health. The rate of glycolysis is one metabolic factor impacting cellular physiology and pathology. For instance, cancer cells exhibit a higher rate of glycolysis compared to healthy counterparts. Since lactate is produced in the course of glycolysis and subsequently released into the extra-cellular space, it acidifies the surrounding medium. The so-called extracellular acidification is hence used to monitor glycolytic flux.

A second metabolic parameter is the consumption of molecular oxygen (O2) as it reflects oxidative phosphorylation. In cancer cells this pathway of energy production is decreased. Furthermore changes in oxygen consumption may indicate mitotoxicity of drugs.

CLARIOstar graph

Monitoring O2 concentrations in samples containing 3D Hep2 cells in response to decreasing atmospheric O2 conditions realized by the CLARIOstar ACU.

Mandel provides probes that are sensitive to either pH or O2 and reliably trace both metabolic processes: glycolysis and respiration. Both probes can be used simultaneously, assuring maximal information output from your experiment.

The CLARIOstar microplate reader is the ideal tool to measure these metabolic assays as it easily detects the probes and can be equipped with an Atmospheric Control Unit (ACU). This accessory allows working at desired oxygen and carbon dioxide concentrations that resemble physiological conditions. It further enables to work at steady O2 and CO2 concentrations, a requirement for determination of extracellular acidification (ECA) and oxygen consumption rate (OCR).

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