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Molecular Probes™ Vybrant™ CFDA SE Cell Tracer Kit
Description
Superior Fluorescent Staining
Successful tracing of cells requires an extremely bright dye to distinguish fluorescently labeled cells from auto-fluorescence. The intense fluorescent staining provided by Vybrant™ CFDA SE Cell Tracer Kit enables the visualization of labeled cells for several days post stain, making it an ideal stain for cell tracing studies based on dye dilution.
Long-Term Signal Retention
Vybrant™ CFDA SE dye easily crosses the plasma membrane and covalently binds inside cells where the stable, well-retained fluorescent dye is designed to provide a consistent signal, even after several days in a cell culture environment.
Non-Toxic
Vybrant™ CFDA SE dye binds covalently to all free amines on the surface and inside of cells and shows little cytotoxicity, with minimal observed effect on the proliferative ability or biology of cells. Researchers have used CFDA SE labeling to show that transplantable hematopoietic cells proliferate in vitro in response to stimulation by a growth factor cocktail.
Simple, Robust Staining Protocol
The Vybrant™ CFDA SE Cell Tracer Kit contains convenient single-use vials of dry dye to permit small-scale experiments without preparing excess quantities of dye, thereby extending the shelf life of the dye not dissolved in solution. A stock solution is prepared by dissolving the contents of a vial in anhydrous DMSO prior to use. To stain 1 mL of cells in protein-free medium, 1μl of this stock solution is typically used. Cells should be stained for 20 minutes at room temperature with gentle agitation. A brief wash with complete medium will then quench any dye remaining in solution.
Specifications
Specifications
| Content And Storage | Contains 10 vials of 5(6)-CFDA, SE (CFSE, 500 μg per vial), and 1 vial of DMSO (1.3 mL). Store in freezer (-5 to -30°C). |
| Excitation Wavelength Range | 492⁄517 |
| Dye Type | CFDA SE (CFSE) |
| Format | Tube(s), Slide(s) |
| For Use With (Equipment) | Fluorescence Microscope, Flow Cytometer |
| Quantity | 1 Kit |
| Detection Method | Fluorescence |
| Form | Solid |
| Product Line | Vybrant |
| Shipping Condition | Room Temperature |
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Frequently Asked Questions (FAQs)
This dye will bind to proteins via primary amines. Too high of a concentration can lead to cell toxicity or unintended modification of cell function. The solution is to reduce the concentration and/or staining time, or to go with a non-protein-binding tracking reagent, such as Qtracker cell labeling reagents.
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.
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.
This is not recommended. When these stains bind to DNA and RNA, they may affect the normal function of the nucleic acids, disrupting transcription, as well as replication. Other reagents, such as CellTracker dyes or Qtracker reagents are more optimized for tracking without disrupting normal activity. If a nuclear label is still desired, though, and the cells are mammalian and non-hematopoietic, CellLight nuclear reagents can transiently transfect cells to express GFP or RFP on a nuclear-expressing protein for up to several days without affecting function.
Please see this Web link (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/cell-tracking.html) to help you choose the right option for your application. Start by planning how long you want to track your cells, then consider the mechanism of binding. Calcein dyes are very uniform in label and are good for short-term cell migration, but may be rapidly effluxed from some cell types. Lipophilic cyanine dyes, such as DiI, DiO, and similar dyes label cell membranes, don’t disrupt function, and can last longer, but have the potential to cross to other cells if membranes fuse. They are also lost upon permeabilization. CellTracker dyes are better for longer-term labeling, as they possess a mildly reactive chloromethyl moiety that allows covalent binding to cellular components. CFDA SE also covalently binds to cellular components. With all the reagents, their retention within cells is dependent upon the rate of cell division and the inherent properties of the cell (active efflux, membrane and protein turnover rates, etc.) and reagents that allow for covalent attachment exhibit longer retention than those that do not.
The longest-lasting and brightest options are the Qtracker reagents, which are taken up through endocytosis. These are so bright individual quantum dots can be detected, and are also robust enough to survive not only fixation and permeabilization, but even the heat and solvents used in paraffin processing.
For Research Use Only. Not for use in diagnostic procedures.
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