Chromeo™ Live Cell Mitochondrial Staining Kit
bright fluorescent mitochondrial stain for living cells
The Chromeo™ Live Cell Mitochondrial Staining Kit utilizes a proprietary, non-toxic, water-soluble, membrane-permeable dye that rapidly diffuses into live or fixed cells. In live cells it stains mitochondria with low background and little or no staining of other organelles. When fixed cells are stained, internal membranes and nucleoli become additional targets of the dye.
Long-term cellular labeling
In addition to serving as a mitochondrial stain, the Chromeo™ Live Cell Mitochondrial Stain can be used as a long-term cell label. Because of the excellent retention of the stain in the cell and its low toxicity, it is an ideal tool for long-term labeling of cells and cellular tracking.
Figure 1: Staining of living HeLa cells with Chromeo™ Live Cell Mitochondrial Staining Kit.
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The Chromeo™ Live Cell Mitochondrial Staining Kit features a unique dye that is not fluorescent until after it has reacted with an amino group, which induces a structural change that results in bright red fluorescence. The dye diffuses passively through the membrane of living cells, and localizes at the mitochondria where reaction and activation occur. The fluorescent dye shows similar properties in fixed cells. In addition to mitochondria, the kit stains intracellular membranes and nucleoli when the cells are first treated with fixation reagents. A long Stokes shift separates the excitation wavelength of 503 nm from the emission maximum at 610 nm. This long Stokes shift separates the fluorescence of the stain from any auto-fluorescence background of living cells. To selectively stain the nuclei of the cells, the blue fluorescent Hoechst stain is also included in the kit.
Chromeo Live Cell Mitochondrial Stain advantages
- Water soluble fluorescent live and fixed cell stain
- No background fluorescence of the free dye
- Low toxicity
- Easy to use
- Long Stokes shift eliminates background of cellular autofluorescence
- Hoechst staining of the nucleus
- Enables long-term labeling experiments
Overview of mitochondria
Mitochondria play an important role within living cells as they are involved in a variety of cellular processes. In addition to their “main” function to generate cellular energy, mitochondria are involved in regulation of cell growth and differentiation; they also play a critical role in the induction of apoptosis, and are part of signaling cascades such as calcium signaling. The structural characteristics of mitochondria are variable and depend on the organism, the cell type and the metabolic state of the individual cell. Cell cycle-dependent changes in mitochondrial morphology have also been reported.
Chromeo™ Live Cell Mitochondrial Stain can be excited between 470 nm and 550 nm (maximum at 503 nm), enabling the use of common lasers or other light sources. A principal characteristic of the dye is its long Stokes shift of about 100 nm with an emission maximum at 610 nm. To visualize the mitochondria in combination with the Hoechst nuclear stain, a commonly used Cy3 filter set (550-580 nm/590-650 nm) will separate the fluorescent spectra. To detect the Hoechst nuclear stain, a standard DAPI filter set (370-410 nm/435-485 nm) can be used.
Contents & Storage
- 1 vial of lyophilized Chromeo™ Mitochondrial stain
- 1 vial of Solubilization Buffer
- 2 vials containing Hoechst Nuclear stain solution
Contents guaranteed 6 months when stored at -20°C in the dark.
The number of experiments will depend on the chosen concentration of the stain, which may vary between different cell types. The kit provides sufficient materials to stain up to 3000 wells in 8-well chambered slides, or up to 6000 wells in a 96-well plate.