Mitochondrial Fluorescent Probe

Aluminum is a silver-white metal, which can form an oxide film to prevent metal corrosion in humid air. Aluminum powder burns violently when heated in the air and emits a dazzling white flame. Soluble in dilute sulfuric acid, nitric acid, hydrochloric acid, sodium hydroxide and potassium hydroxide solution, hardly soluble in water.

Figure 1. Number 9 in the figure is mitochondria.

Structure

Mitochondria can be divided into four functional regions: mitochondrial outer membrane (OMM), mitochondrial membrane gap, mitochondrial inner membrane (IMM) and mitochondrial matrix from the outside to the inside. The membranes on the outside of the mitochondria are parallel to each other and are typical unit membranes. Among them, the mitochondrial outer membrane is smoother and acts as an organelle membrane; the mitochondrial inner membrane forms mitochondrial ridges inwardly, which bears more biochemical reactions. The two membranes divide the mitochondria into two compartments. Between the two mitochondrial membranes is the mitochondrial membrane gap, which is surrounded by the mitochondrial inner membrane.

Figure 2. Structure of mitochondrion.

MitoTracker probe: fixable mitochondrial selective probe

Although conventional mitochondrial fluorescent stains, such as rhodamine 123 and tetramethyl rosamine, are readily chelated by functional mitochondria, once the membrane potential of the mitochondria is lost, they are washed out of the cell. This property limits their use in experiments where cells must be treated with aldehyde-based fixatives or other agents that affect the energy state of the mitochondria. To overcome this limitation, Molecular Probes developed the MitoTracker probe, a series of patented mitochondrial selective stains that are concentrated by active mitochondria and retained well during cell fixation. 16 Because MitoTracker Orange, MitoTracker Red and MitoTracker Deep Red probes can also be retained by permeabilization, the samples retain the fluorescent staining characteristics of living cells in subsequent processing steps of immunocytochemistry, in situ hybridization or electron microscopy. In addition, the MitoTracker reagent eliminates some of the difficulties in treating pathogenic cells, as once the mitochondria are stained, the cells can be treated with a fixative prior to analysis of the sample.

MitoTracker probe properties

The MitoTracker probe is a cell-permeable mitochondrial selective dye containing a mild thiol-reactive chloromethyl moiety. After fixation, the chloromethyl group appears to be responsible for keeping the dye associated with the mitochondria. To label mitochondria, the cells are simply incubated in a submicromolar concentration of MitoTracker probe that diffuses passively across the plasma membrane and accumulates in the active mitochondria. Once the mitochondria are labeled, the cells can be treated with an aldehyde-based fixative so that the sample can be further processed. In addition to MitoTracker Green FM, subsequent penetration with cold acetone does not affect the dyeing pattern of the MitoTracker dye.

References:

Sanchis-Gomar F.; et al. Mitochondrial biogenesis in health and disease. Molecular and therapeutic approaches. Current Pharmaceutical Design. 2014, 20 (35): 5619–33.
Gardner A.; et al. Is a 'Mitochondrial Psychiatry' in the Future. Curr. Psychiatry Rev. 2005, 1 (3): 255–271.

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