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Invitrogen™ CellTracker™ Fluorescent Probes
Beskrivning
Cell movement and location studies require specialized probes that are nontoxic to living cells and well retained, allowing for multigenerational tracking. The CellTracker fluorescent probes are available in a range of fluorescent colors to match instrument lasers and filters, and to accommodate co-staining with antibodies or other cell analysis probes. These dyes are excellent tools for monitoring cell movement, location, proliferation, migration, chemotaxis, and invasion.
Features of the CellTracker dyes include:
- Easy to use—remove culture media, add dye, incubate 15-45 minutes, and image cells
- Excellent retention—fluorescent signal retention of >72 hours (typically three to six generations)
- Ideal tracking dyes—monitor cell movement, location, proliferation, migration, chemotaxis, and invasion
- Low cytotoxicity—does not affect viability or proliferation
- Cell Versatility– works with cells in suspension and adherent cells
The CellTracker fluorescent probes have been designed to freely pass through cell membranes; however, once inside the cell are transformed into cell-impermeant reaction products. The CellTracker fluorescence probes (except for CellTracker Deep Red) contain a chloromethyl or bromomethyl group that reacts with thiol groups, utilizing a glutathione S-transferase–mediated reaction. In most cells, glutathione levels are high (up to 10 mM) and glutathione transferase is ubiquitous. CellTracker Deep Red probe contains a succinimidyl ester reactive group, which reacts with amine groups present on proteins.
After conversion to impermeant versions, the CellTracker fluorescent probes are well retained in living cells through several generations. The probes are transferred to daughter cells, but are not transferred to adjacent cells in a population. Cells loaded with the CellTracker fluorescent probes display fluorescence for at least 72 hours and exhibit ideal tracking dye properties—they are stable, nontoxic at working concentrations, well retained in cells, and brightly fluorescent at physiological pH. Additionally, several CellTracker fluorescent probes with various excitation and emission spectra are available allowing for multiplexing in fluorescence imaging. The CellTracker dyes are retained with fixation and permeabilization, enabling their use with antibodies for fluorescence immunostaining applications.
Spectral characteristics of the fluorescent CellTracker probes:
- CellTracker Blue CMAC—353/466 nm
- CellTracker Blue CMF2HC—371/464 nm
- CellTracker Blue CMHC—372/470 nm
- CellTracker Violet BMQC—415/516 nm
- CellTracker Green CMFDA—492/517 nm
- CellTracker Green BODIPY™—522/529 nm
- CellTracker Orange CMTMR—541/565 nm
- CellTracker Orange CMR—548/576 nm
- CellTracker Red CMTPX—577/602 nm
- CellTracker Deep Red—630/660 nm
Fluorescent CellTracker reagents include the blue-fluorescent chloromethyl derivatives of amino-, hydroxy-, and difluorohydroxycoumarin (CMAC, CMHC and CMF2HC), the green-fluorescent chloromethyl derivatives of fluorescein diacetate (CMFDA) and a BODIPY™ dye, the orange-fluorescent CMTMR and CMRA, and the red-fluorescent CMTPX. CellTracker Blue CMAC, CMHC, and CMF2HC, CellTracker Violet, the violet-fluorescent bromomethyl derivative of coumarin (BMQC), CellTracker Green BODIPY™, CellTracker Orange CMTMR, and CellTracker Red CMTPX do not require enzymatic cleavage to activate their fluorescence, whereas the green CMFDA and orange CMRA do require enzymatic cleavage. The impermeable reaction products of the chloromethyl or bromomethyl coumarins have excellent retention, strong fluorescence, and relatively uniform cytoplasmic staining, making these derivatives potentially useful for correcting motion artifacts in imaging. CMFDA is colorless and non-fluorescent until cytosolic esterases cleave off the acetates, releasing a brightly fluorescent product.
In addition to bright fluorescence and excellent retention, the CellTracker fluorescent probes do not contribute to cytotoxicity.
Beställningsinfo
Shipping Condition: Room Temperature
Specifikationer
Specifikationer
| Color | Green |
| Content And Storage | Store in freezer (-5°C to -30°C). |
| Excitation Wavelength Range | 492 nm |
| Dye Type | CellTracker Green CMFDA |
| Quantity | 1 mg |
| Description | CellTracker™ Green CMFDA Dye, 1 mg |
| Emission | 517 nm |
| Form | Dry Powder |
| Product Line | CellTracker |
| Reagent Type | Cell Tracker Compounds, Cell Labeling Reagents |
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Frequently Asked Questions (FAQs)
Calcein, AM and FDA (fluorescein diaceate) are examples of some dyes used for this application. Since these dyes are not incorporated or covalently attached to any cellular components, they may have a short retention time as some cell types may actively efflux the dye out of the cells. The CellTracker and CellTrace dyes include either a mild thiol-reactive chloromethyl group or amine-reactive succinnimidyl ester group to allow for covalent binding to cellular components, providing for better retention. As with any reagent, one should empirically determine retention times for the cell type used.
DAPI is considered a semi-permeant/impermeant nucleic acid stain. Staining of nucleic is dependent upon the cell line in its performance. Some cell lines will label with DAPI, others not at all, and others label inconsistenly. Instead, we recommend using either Hoechst 33342 or Hoechst 33258, which have the same wavelength and binding mode as DAPI (at the A-T minor groove) but are readily cell-permeant.
We do not recommend doing this. DAPI is considered to be a semi-permeant/impermeant nucleic acid stain. DAPI staining of live cells may be inconsistent. It is best used as a counterstain for fixed samples. Other cell permeable nucleic acid stains, such as Hoechst or the SYTO dyes may affect cellular function.
For mammalian cells, we recommend using the CellLight Nucleus transduction reagents, available in CFP, GFP and RFP. With these reagents, the cells are transduced overnight in a single labeling step and the next day the nuclei will fluoresce. The label may be retained for 3-5 days and should not affect cell function. Cytoplasmic cell tracking dyes such as the CellTracker dyes may also be used.
Calcein, AM is a good choice for cell tracking and as a general cytoplasmic stain. However, it doesn't bind to anything and may be actively pumped out of the cells within a couple hours, which is likely what happened. The retention of Calcein within live cells is dependent upon the inherent properties of the cell type and culture conditions.
For long-term imaging, you may wish to consider a reactive cytoplasmic stains such as CFDA, SE or the CellTracker and CellTrace dyes.
Yes, the CellTracker dyes react with any accessible thiol part of the protein and can be fixed. However, some CellTracker dyes may be attached to small metabolites that can leak from the cell following permeabilization. This can result in decreased fluorescence.
For Research Use Only. Not for use in diagnostic procedures.
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