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| Product Name | CFDA-SE Cell Proliferation Tracing Fluorescent Probe |
| Catalog No. | RESK-0529 |
| Description | CFDA, SE (carboxyfluorescein diacetate, succinimidyl ester) is a cell tracing dye for fluorescent labeling of living cells. It is suitable not only for in vitro cell proliferation assays but also for tracking cell division and proliferation in vivo. CFDA, SE is a derivative of fluorescein diacetate (FDA). It is membrane-permeant and intrinsically non-fluorescent. After passively crossing the membrane of living cells, it is hydrolyzed by cytosolic esterases to carboxyfluorescein succinimidyl ester (CFSE), which emits intense green fluorescence, cannot cross the cell membrane, and is well retained intracellularly. CFSE also spontaneously and irreversibly binds intracellular amino groups, covalently coupling to cellular proteins, while excess uncoupled CFDA, SE passively diffuses back into the culture medium and is removed by subsequent washing steps. Labeled non-dividing cells display very stable fluorescence, with stable labeling lasting for months, making the dye highly suitable for cell population analysis. Fluorescence labeling with CFDA, SE is highly homogeneous, superior to previously used cell tracking fluorescent probes such as PKH26, and the fluorescence is partitioned more evenly among daughter cells after division. During proliferation, the CFSE signal is distributed equally to the two daughter cells, reducing the intensity to half that of the parent cell. Using flow cytometry (FL1 channel), undivided cells and cells that have divided once (1/2 intensity), twice (1/4), three times (1/8), or more can be distinguished; eight or more divisions can be detected. Labeled cells can be used for in vitro and in vivo proliferation studies without staining neighboring cells. CFDA, SE is most commonly used for proliferation analysis of lymphocytes and is also applicable to fibroblasts, natural killer cells, hematopoietic progenitor cells, and other cell types. Labeled cells show green fluorescence (Ex = 494 nm, Em = 521 nm). Besides flow cytometric proliferation analysis, a fluorescence microplate reader can be used to quantify viable cell numbers, and fluorescence microscopy can be used for homogeneous cell tracing observation. This product is supplied as a powder and must be dissolved in DMSO to prepare a stock solution before use. |
| Intended Use | Fluorescent labeling and tracing of living cells for cell proliferation studies in vitro and in vivo. |
| Principle / Technology | CFDA, SE is a membrane-permeant, non-fluorescent fluorescein diacetate derivative. Intracellular esterases hydrolyze it to CFSE, a membrane-impermeant, brightly green-fluorescent product that covalently and irreversibly couples to intracellular proteins through amino groups. Because the fluorescence is divided equally between daughter cells at each division, the number of cell divisions can be deduced from the stepwise halving of fluorescence intensity. |
| Detection Method | Flow cytometry (FL1 channel); fluorescence microscopy; confocal microscopy; fluorescence microplate reader. Ex = 494 nm, Em = 521 nm. |
| Sample Type | Living cells or tissues (direct staining); cells or tissues fixed with 4% paraformaldehyde. Commonly used for lymphocytes; also applicable to fibroblasts, natural killer cells, and hematopoietic progenitor cells. |
| Performance Range / Specifications | Detects up to eight or more cell divisions by stepwise halving of fluorescence intensity. Stable labeling of non-dividing cells can last for months. |
| Specificity | Esterase-dependent labeling of living cells; does not stain neighboring cells within the population. For bacterial proliferation analysis, the cell wall must first be removed before staining. |
| Reaction Conditions / Protocol | Prepare a stock solution in DMSO before use. Living cells or tissues can be stained directly; fixed samples are typically fixed with 4% paraformaldehyde. Excess uncoupled probe is removed by washing steps after labeling. |
| Components / Formulation | Single-component fluorescent probe supplied as an off-white solid powder. |
| Storage Conditions | Store at -20 °C in a dry place, protected from light. Avoid repeated freeze-thaw cycles. |
| Shelf Life | 2 years under recommended storage conditions. |
| Package Specifications | 25 mg |
| Product Form | Off-white solid powder. |
| Purity | >=94% (HPLC) |
| Molecular Weight | 557.46 |
| Source / Origin | Chemical synthesis. CAS No. 150347-59-4; molecular formula C29H19NO11; English synonym: 5-(and 6)-Carboxyfluorescein diacetate, succinimidyl ester. |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Stable for 2 years when stored dry at -20 °C protected from light; avoid repeated freeze-thaw cycles. Fluorescence of labeled non-dividing cells remains stable for months. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with fluorescence microscopes, confocal microscopes, microplate readers, and flow cytometers. |
| Recommended Buffer System | Dissolve in DMSO to prepare the stock solution before use. |
| Application Notes / Precautions | For fixed cells or tissues, fixation with 4% paraformaldehyde is recommended; other inappropriate fixatives can cause high fluorescence background or poor staining. For bacterial proliferation analysis, the cell wall should be removed before staining. Avoid repeated freeze-thaw cycles of the product. |
For research use only, not for clinical use.
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