Method of measuring mitochondrial permeability transition pore opening Image-IT LIVE Mitochondrial Transition Pore Assay Please see the table of MitoTracker probes for Imaging and Flow Cytometry below. Only a small selection of the end point assays for detecting mitochondria membrane potential are fixable. Loss of mitochondrial calcein fluorescence due to mitochondrial permeability transition. Measure the increasing fluorescence of a calcium indicator with increasing calcium concentration in the mitochondria.

Measure the increasing fluorescence of a superoxide detection reagent that accumulates in mitochondria in a membrane potential-dependent fashion where it is selectively oxidized and subsequently binds nucleic acids. Measure fluorescence of reagents that accumulate in active (healthy) mitochondria with intact membrane potentials.

Irreversible detection of the mitochondrial permeability transition (MPT) subsequent to dysregulation of the MPT pore. Irreversible detection of mitochondrial calcium in live cells. Irreversible detection of mitochondrial superoxide production in live cells. Changes in the membrane potential, along with decreases ATP to ADP ratios, increased mitochondrial matrix calcium levels, oxidative stress, and release of cytochrome c into the cytosol, are all presumed to be associated with the mitochondrial permeability transition, resulting in disruption of ions and small molecule homeostasis via the mitochondrial permeability transition pore (MPTP).Ĭells with healthy, metabolically active mitochondria.ĭetection of dynamic changes in relative mitochondria membrane potential (reversible detection).Įnd point assays for detecting relative mitochondria membrane potential at a specific timepoint (irreversible detection).

Therefore, the membrane’s depolarization is a good indicator of mitochondrial dysfunction, which is increasingly implicated in drug toxicity ( 2-6). The inner mitochondrial membrane potential is essential in Ca 2+ uptake and storage, reactive oxygen species generation, detoxification, and most importantly, the synthesis of ATP by oxidative phosphorylation ( 1). This mitochondrial disruption includes changes in the membrane potential - a central feature of mitochondrial health, and alterations to the oxidation–reduction potential of the mitochondria. A feature of the early stages of programmed cell death is the disruption of mitochondrial function.

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