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| Product Name | CFDA SE Cell Proliferation and Cell Tracking Kit |
| Catalog No. | RESK-0196 |
| Description | The CFDA SE Cell Proliferation and Cell Tracking Kit is designed for fluorescent cell tracing and proliferation analysis based on CFDA, SE (carboxyfluorescein diacetate, succinimidyl ester). The kit contains CFDA, SE powder together with the solvent required for stock solution preparation and a dedicated cell staining buffer, greatly simplifying assay setup. CFDA, SE is a membrane-permeant, non-fluorescent compound. After passively crossing the membrane of living cells, it is hydrolyzed by intracellular esterases to carboxyfluorescein succinimidyl ester (CFSE), which emits intense green fluorescence (Ex = 494 nm, Em = 521 nm), can no longer cross the cell membrane, and is well retained inside the cell. CFSE also spontaneously and irreversibly reacts with intracellular amino groups, covalently coupling to cellular proteins, while excess uncoupled CFDA, SE diffuses back into the culture medium and is removed by subsequent washing steps. Labeled non-dividing cells show extremely stable fluorescence, with stable labeling lasting for months, making the kit highly suitable for cell population analysis. Labeling is more homogeneous than with earlier cell tracking probes such as PKH26, and fluorescence is distributed evenly to daughter cells. During proliferation, the CFSE fluorescence is divided equally between the two daughter cells, so fluorescence intensity is halved with each generation. Cells that have divided once (1/2 intensity), twice (1/4), three times (1/8), or more can be resolved by flow cytometry (FL1 channel), and eight or more divisions can be detected. Labeled cells can be used for both in vitro and in vivo proliferation studies without staining neighboring cells. The most common application is lymphocyte proliferation analysis; fibroblasts, natural killer cells, hematopoietic progenitor cells, and other cell types can also be analyzed. In addition to flow cytometry, labeled cells can be quantified on a fluorescence microplate reader or observed by fluorescence microscopy. |
| Intended Use | Fluorescent tracing and proliferation analysis of living cells in vitro and in vivo, including lymphocytes, fibroblasts, natural killer cells, and hematopoietic progenitor cells. |
| Principle / Technology | CFDA, SE is membrane-permeant and non-fluorescent. Inside living cells, cytosolic esterases convert it to CFSE, a brightly green-fluorescent, membrane-impermeant product that covalently and irreversibly couples to intracellular proteins via amino groups. Because the fluorescence is partitioned equally between daughter cells at each division, the fluorescence intensity is halved per generation, allowing individual division rounds to be resolved by flow cytometry. |
| Detection Method | Flow cytometry (FL1 channel; Ex = 494 nm, Em = 521 nm); fluorescence microplate reader for quantitation of viable cell numbers; fluorescence microscopy for cell tracing observation. |
| Sample Type | Living cells in suspension or culture, including lymphocytes, fibroblasts, natural killer cells, and hematopoietic progenitor cells; suitable for in vitro and in vivo proliferation studies. |
| Performance Range / Specifications | Resolves up to eight or more cell divisions based on stepwise halving of CFSE fluorescence intensity. Labeling of cells is generally completed within 15 min; the optimal labeling time should be determined empirically for each cell type. Calculated at a labeling volume of 2 mL per sample, the kit formats support 500 and 1000 assays, respectively. |
| Specificity | Esterase-dependent labeling of living cells; labeled cells do not stain neighboring cells, and fluorescence is transferred to daughter cells rather than to adjacent cells in the population. |
| Reaction Conditions / Protocol | Cell labeling is generally completed within 15 min; the optimal labeling time should be optimized for each cell type. The standard labeling volume is 2 mL per sample. Excess uncoupled probe is removed by washing steps after labeling. |
| Components / Formulation | CFDA, SE powder Solvent for preparation of the CFDA, SE stock solution Cell staining buffer for cell labeling |
| Storage Conditions | Store at -20 °C in a dry place, protected from light. |
| Shelf Life | 2 years under recommended storage conditions. |
| Package Specifications | 500 T 1000 T |
| Product Form | Kit containing CFDA, SE powder, solvent, and cell staining buffer. |
| Concentration | Solid content is approximately 1.4 mg per vial; after dissolution in the supplied solvent, the concentration is approximately 2.8 mg/mL. |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Stable for 2 years when stored dry at -20 °C protected from light. Fluorescence of labeled non-dividing cells remains stable for months, enabling long-term cell population analysis. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with flow cytometers (FL1 channel), fluorescence microplate readers, and fluorescence microscopes. |
| Recommended Buffer System | The kit provides a dedicated cell staining buffer for the labeling reaction. |
| Application Notes / Precautions | Labeling with CFDA, SE is more homogeneous than with earlier cell tracking probes such as PKH26, and daughter cells receive a more even fluorescence distribution. The optimal labeling time varies between cell types and should be determined empirically. Store protected from light. |
For research use only, not for clinical use.
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