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Invitrogen™ MitoTracker™ Dyes for Mitochondria Labeling
Description
Rosamine & carbocyanine-based staining dyes MitoTracker Orange CMTMRos, Red CMXRos, Orange CM-H2TMRos, Red CM-H2XRos, Red FM, Green FM, & Deep Red FM enable mitochondria visualization with fluorescent imaging. These mitochondria dyes are specially packaged in vials of 50 μg each to create freshly reconstituted dyes without the impact of freeze-thaw cycles to aid in a simplified sample prep for cell analysis workflow.
Rosamine-based MitoTracker dyes for mitochondria labeling
Cell-permeant MitoTracker probes stain active mitochondria in live cells for labeling and localization in fluorescent cell imaging. Our rosamine-based mitochondrial staining dyes include MitoTracker Orange CMTMRos, a derivative of tetramethylrosamine, and MitoTracker Red CMXRos, a derivative of X-rosamine. MitoTracker Orange CM-H2TMRos and MitoTracker Red CM-H2XRos are reduced, nonfluorescent versions of the rosamine-based MitoTracker dyes that fluoresce upon oxidation. The fluorescent signal from the rosamine-based MitoTracker dyes is retained in the mitochondria even after aldehyde fixation and detergent permeabilization, making these mitochondria dyes flexible for many workflows including applications that require subsequent processing such as immunocytochemistry or in situ hybridization. MitoTrackerRed and Orange are well suited for multicolor labeling experiments because their fluorescence is well resolved from the green fluorescence of other probes.
Carbocyanine-based MitoTracker dyes for mitochondria labeling
Cell-permeant MitoTracker dyes stain active mitochondria in live cells for mitochondrial labeling and localization in fluorescent cell imaging. The carbocyanine-based dyes MitoTracker Green FM and MitoTracker Red FM accumulate in active mitochondria but are not well-retained in mitochondria after aldehyde fixation. The carbocyanine-based MitoTracker Deep Red FM stains active mitochondrial in live cells and is well-retained in mitochondria after aldehyde fixation and subsequent permeabilization with detergents for applications that require subsequent processing such as immunocytochemistry or in situ hybridization. MitoTracker Red FM and MitoTracker Deep Red FM are well suited for multicolor labeling experiments because their red fluorescence is well resolved from the green fluorescence of other probes.
Benefits of using MitoTracker mitochondria labeling dyes
MitoTracker dyes are provided in vials of 50 μg of lyophilized powder ready for reconstitution. To label mitochondria, live cells are simply incubated with the MitoTracker probe of your choice. The mitochondrial staining dyes passively diffuse across the plasma membrane and accumulate in active mitochondria. The MitoTracker dyes are offered in a range of wavelengths and can be used for mitochondrial localization in multicolor experiments.
Conventional fluorescent stains such as tetramethylrosamine and rhodamine 123 are readily sequestered by active mitochondria and are reversible in dynamic membrane potential measurements as they easily wash out of cells upon loss in membrane potential. In contrast, MitoTracker dyes contain a mildly thiol-reactive chloromethyl moiety so that mitochondrial staining is retained if the mitochondrial membrane potential is lost, allowing many of the MitoTracker dyes to be retained during cell fixation.
Throughout the cell life cycle, mitochondria use oxidizable substrates to produce an electrochemical proton gradient across the mitochondrial membrane (whose potential is negative), resulting in ATP production. MitoTracker dyes are ideal probes for mitochondria staining in experiments studying the cell cycle or processes such as apoptosis and other end point assays. MitoTracker dyes are also available for flow cytometry applications (Cat. No. M46750, M46751, M46752, and M46753).
Related products for mitochondrial membrane potential
For studying dynamic mitochondria membrane potential specifically, we recommend using JC-1 (cationic carbocyanine dye, Cat. No. T3168) or TMRM (tetramethyl rhodamine methyl ester, Cat. No. I34361) dyes. The MitoProbe JC-1 Assay Kit (Cat. No. M34152) contains the JC-1 dye in addition to the potent mitochondrial membrane-potential disrupter, CCCP, which depolarizes the mitochondrial membrane. These reagents can provide compensatory controls to correctly compensate green-to-red fluorescence ratio. TMRM is used for the detection of dynamic measurement of mitochondrial membrane potential.
Order Info
Shipping Condition: Room Temperature
Specifications
Specifications
| Color | Red |
| Content And Storage | Store in freezer -5°C to -30°C and protect from light. |
| Excitation Wavelength Range | 579/599 nm |
| Format | Special packaging |
| For Use With (Equipment) | Fluorescence Microscope, Flow Cytometer, Microplate Reader |
| Quantity | 20 x 50 μg |
| Detection Method | Fluorescence |
| Description | MitoTracker™ Red CMXRos |
| Product Line | MitoTracker |
| Shipping Condition | Room Temperature |
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Frequently Asked Questions (FAQs)
This is typically a result of using too high of a concentration of the Mitotracker dye. Most organic dyes are used in the low micromolar range. The MitoTracker dyes are used at a much lower concentration, around 50–200 nanomolar. Higher concentrations can cause background fluorescence and non-mitochondrial staining.
Yes, it has been done. A literature search will find numerous examples. However, be aware that plate readers are typically less sensitive than microscopes or flow cytometers, which means that the degree of change in fluorescence may not be as sensitive or easily detectable with a plate reader. Be sure to optimize label times and concentrations carefully.
Yes, this is possible because MitoTracker Red CMXRox dye is retained upon fixation. See Table 2 in this manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp07510.pdf).
The Mitotracker dyes should be imaged soon after staining because over time, those dyes can be toxic.
-Calcium flux: Each of the Oregon Green calcium indicators binds intracellular calcium with increasing affinity, providing a sensitivity range to match many applications. Oregon Green probes emit green fluorescence at resting levels of Ca2+ and increase their fluorescence intensity 14-fold with increasing Ca2+ concentration. The cell-permeant formulation (Cat. No. O6807) can be loaded in cell media and is compatible with flow cytometry.
-Rhodamine-based calcium indicators comprise a range of probes for large or small changes in Ca2+ concentration. They exhibit a 50-fold increase in fluorescence upon calcium binding and offer a range of wavelengths that can be used in conjunction with GFP or green-fluorescent dyes for multiplexing. Rhod-2, AM (Cat. No. R1245MP), in particular, localizes to mitochondria and can be used with flow cytometry.
-Membrane potential: A distinctive feature of the early stages of apoptosis is the disruption of the mitochondria, including changes in membrane and redox potential. We offer a range of products specifically designed to assay mitochondrial membrane potential in live cells by flow cytometry, with minimal disruption of cellular function. The MitoProbe family of mitochondrial stains (Cat. Nos. M34150, M34151, and M34152) provide quick, easy, and reliable flow cytometric detection of the loss of mitochondrial membrane potential that occurs during apoptosis. MitoTracker dyes (Cat. Nos. M7510 and M7512) are membrane potential-dependent probes for staining mitochondria in live cells. The staining pattern of MitoTracker dyes is retained throughout subsequent flow cytometry immunocytochemistry, DNA end labeling, in situ hybridization, or counterstaining steps. The Mitochondrial Permeability Transition Pore Assay (Cat. No. M34153) provides a more direct method of measuring mitochondrial permeability transition pore opening than assays relying on mitochondrial membrane potential alone. The mitochondrial permeability transition pore (MPTP) is a non-specific channel formed by components from the inner and outer mitochondrial membranes, and appears to be involved in the release of mitochondrial components during cell death.
-Phagocytosis: In phagocytosis, cells internalize particulate matter such as microorganisms, and this process is important for immune responses and during the clearance of apoptotic cells. Probes for studying phagocytosis include BioParticles indicators—bacteria and yeast labeled with fluorescent dyes.
-Tracking phagocytosis using a quench/wash-based assay can report on simple uptake, or a pH indicator can be used to monitor stages in the pathway. We have no-wash assays labeled with pHrodo Red or Green (Cat. Nos. A10010, P35361, P35364, P35365, P35366, and P35367) and no-wash assays for whole blood (Cat. Nos. A10025, A10026, P35381, and P35382), all suitable for flow cytometry.
-pH changes: Sensitive pH determinations can be made in a physiological range using either fluorescent intensity or ratiometric measurements. pHrodo dyes (Cat. Nos. P35373 and P35372) provide signal intensity modulation from pH 2 to pH 9 and with a choice of fluorescent wavelengths. Tracking internalization of fluorescent dextran is a routine method for analyzing pH changes in cellular compartments. Dextran conjugates of pHrodo dyes (Cat. Nos. P35368 and P10361) provide the most complete solution by allowing discrimination of vesicles from early endosomes to lysosomes, with no quench or wash required.
-Reactive oxygen species: Cells that are environmentally stressed usually contain greatly increased levels of reactive oxygen species (ROS). CellROX reagents are fluorogenic probes developed for the detection and quantitation of ROS in live cells. These cell-permeant reagents are non-fluorescent or very weakly fluorescent in the reduced state; however, when oxidized, they become brightly fluorescent and remain localized within the cell. We offer CellROX Green (Cat. No. C10492), CellROX Orange (Cat. No. C10493), and CellROX Deep Red (Cat. No. C10491) Assay Kits validated for flow cytometry.
For Research Use Only. Not for use in diagnostic procedures.
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