Cayman Chemical Mito Stress Test (ACU Format)

Cayman Chemical, in collaboration with BMG LABTECH, has developed a novel assay kit to measure mitochondrial function under a dynamic range of oxygen concentrations. The Mito Stress Test (ACU Format) is designed to measure oxygen consumption rate (OCR) and extracellular acidification (ECA) simultaneously and in real time under tightly controlled atmospheric conditions using open-flow respirometry.

The assay is designed to work with microplate readers equipped with an atmospheric control unit (ACU) such as BMG LABTECH’s multifunctional CLARIOstar microplate reader, which is equipped with dual injection ports and time-resolved fluorescence detection. The ACU integrated into the CLARIOstar reader facilitates measurement of cellular oxygenation, while phosphorescent extracellular O₂ (MitoXpress Xtra) and fluorescent extracellular pH (pH-Xtra) probes provided in the kit enable multiparametric detection of electron transport chain activity and glycolytic flux.

The kit eliminates the hyperoxic conditions under which other cell-based in vitro assays are conducted and provides a more physiologically relevant model, according to the company. Rather than using a typical microchamber, the kit provides an alternative platform that relies on the difference in steady-state oxygen, which allows measurements of OCR and ECA under various, user-controlled oxygen concentrations.

The ACU can fully regulate both O₂ and CO₂ at a range from 0.1–20%, which provides a means to measure mitochondrial function at atmospheric concentrations that are lower than ambient conditions. Predefined scripts automatically control the CLARIOstar, making assay performance and data analysis a plug-and-play and walk-away experience.