CFDA-SE (Synonyms: 5(6)Carboxyfluorescein diacetate succinimidyl ester, 5(6)-CFDA N-succinmidyl ester) |
| Katalog-Nr.GC14056 |
CFDA-SE, mit vollem Namen Carboxycein-Diacetat-Succinimidylester und Zellmembranpermeabilität, ist ein fluoreszierender Farbstoff, der weit verbreitet für die Verfolgung lebensfähiger Zellen oder die Detektion von Zellproliferation eingesetzt wird.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 150347-59-4
Sample solution is provided at 25 µL, 10mM.
CFDA-SE, full name carboxycein diacetate, Succinimidyl Ester, with cell membrane permeability, is a fluorescent dye CFDA-SE widely used for viable cell tracing or cell proliferation detection[1].The fluorescence of CFDA-SE -labeled cells was very uniform and stable, and the fluorescence could be evenly distributed to the two progeny cells [2]. CFDA-SE can deliver strong green fluorescence,Ex=494 nm,Em=521nm.
In Human erythroleukaemic cell line K562, and other cell lines, although all cells are labeled with relatively high fluorescence intensity by CFDA-SE, the efficiency of labeling is highly variable[3].Electron dense vesicles are seen at the ultrastructural level in labeled cells. Discrete vesicular labeling can also be observed in whole cell mounts viewed with fluorescence microscopy. Whole cells retain good label for 6 weeks. CFDA-SE labeling is relatively easy, nontoxic to cells and nonradiocactive[4].
In vitro lymphocyte cell proliferation analysis by CFDA-SE method is a reliable and practical choice for the assessment of mitogenic T lymphocyte responses in yet unclassified PID patients for targeting further genetical analyses[5].When NM is detected by flow cytometry, FL1 detection channel CFDA-SE -labeled cells can be adopted, and fluorescence microscopy can also be used to observe CFDA-SE -labeled cells without staining adjacent cells [6].
CFDA-SE has been reported to be effective in labeling CD34+ cells, CD34+ cells isolated from fetal liver or thymus were labeled with CFDA-SE and were injected into a human thymus grafted subcutaneously in the RAG-2 / IL-2Rγ / mice. One to 4 weeks later the CFDA-SE label was found not only in T cells but also in CD123+/high CD4+CD45RA+ pDC2, indicating that the CD34+ cells can develop into pDC2 within a thymus. In addition to pDC2, CFDA-SE -labeled dendritic cells with a mature phenotype, determined by the cell surface markers CD11c, CD83, and CD80, were found in the injected human thymus graft. pDC2 was not found in the periphery of mice carrying a human thymic graft, indicating that the intrathymic pDC2 failed to emigrate from the thymus[7]. In C57BL/6 mice，CFDA-SE, at 80 times the concentration used for in vitro labeling, was nontoxic and labeled randomly approximately 15% of thymocytes 24 h after injection. The turnover rate of labeled thymic emigrants in the lymph nodes was in the order of 21 days. Thus, CFDA-SE may serve as a powerful tool in relatively long-term migration studies[8].
References:
[1]: Chen JC, Chang ML, et,al. A kinetic study of the murine mixed lymphocyte reaction by 5,6-carboxyfluorescein diacetate succinimidyl ester labeling. J Immunol Methods. 2003 Aug;279(1-2):123-33. doi: 10.1016/s0022-1759(03)00236-9. PMID: 12969553.
[2]: Lyons AB, Blake SJ, et,al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11. doi: 10.1002/0471142956.cy0911s64. PMID: 23546777.
[3]: Wang XQ, Duan XM, et,al. Carboxyfluorescein diacetate succinimidyl ester fluorescent dye for cell labeling. Acta Biochim Biophys Sin (Shanghai). 2005 Jun;37(6):379-85. doi: 10.1111/j.1745-7270.2005.00051.x. PMID: 15944752.
[4]: Gruber HE, Leslie KP, et,al. Optimization of 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester for labeling human intervertebral disc cells in vitro. Biotech Histochem. 2000 May;75(3):118-23. doi: 10.3109/10520290009066489. PMID: 10950173.
[5]: Azarsiz E, Karaca N, et,al. In vitro T lymphocyte proliferation by carboxyfluorescein diacetate succinimidyl ester method is helpful in diagnosing and managing primary immunodeficiencies. J Clin Lab Anal. 2018 Jan;32(1):e22216. doi: 10.1002/jcla.22216. Epub 2017 Apr 6. PMID: 28383134; PMCID: PMC6816938.
[6]: Li X, Dancausse H, et,al. Labeling Schwann cells with CFSE-an in vitro and in vivo study. J Neurosci Methods. 2003 May 30;125(1-2):83-91. doi: 10.1016/s0165-0270(03)00044-x. PMID: 12763234.
[7]: Weijer K, Uittenbogaart CH, et,al. Intrathymic and extrathymic development of human plasmacytoid dendritic cell precursors in vivo. Blood. 2002 Apr 15;99(8):2752-9. doi: 10.1182/blood.v99.8.2752. PMID: 11929763.
[8]: Graziano M, St-Pierre Y, et,al. The fate of thymocytes labeled in vivo with CFSE. Exp Cell Res. 1998 Apr 10;240(1):75-85. doi: 10.1006/excr.1997.3900. PMID: 9570923.
|
Protocol for Cell labeling and counting with CFDA-SE [1]: |
|
|
Cells were counted and stained with CFDA-SE, as follows:
This protocol only provides a guideline, and should be modified according to your specific needs. |
|
|
References: [1]. Urbani S, Caporale R,et,al. Use of CFDA-SE for evaluating the in vitro proliferation pattern of human mesenchymal stem cells. Cytotherapy. 2006;8(3):243-53. doi: 10.1080/14653240600735834. PMID: 16793733. |
|
|
Cell experiment [1]: |
|
|
Cell lines |
Human erythroleukaemic cell line K562, mouse lymphoma cell line YAC-1, human mammary cancer cell line MCF-7 and human melanoma cell line A375 |
|
Preparation method |
The 5 mM CFDA-SE stock in DMSO was diluted to different concentrations (2 μM, 3 μM, 4 μM, 5 μM, 10 μM and 20 μM) in PBS with a total volume of 1 ml. Cells were added to equal volume of CFDA-SE with different concentrations and incubated at 37 C for 5, 6, 7, 8, 10 and 15 min with agitation. |
|
Reaction Conditions |
1, 1.5, 2, 2.5, 5 and 10 µM CFDA-SE |
|
Applications |
CFDA-SE at 2.5 μM stained more than 95% of the cells on all cell lines tested, and dose-dependently increased fluorescence intensity of stained cells. The optimal concentration for K562 and YAC-1 was found to be 2.5 μM, while the optimal concentrations for A375 and MCF-7 were found to be 5 μM and 10 μM, respectively. Within the 6 h experiment period, no cytotoxicity related to CFDA-SE was observed. |
|
Animal experiment [2]: |
|
|
Animal models |
C57BL/6 mice, aged 5 ~ 8 weeks |
|
Preparation method |
Injected CFDA-SE into thymic lobe |
|
Dosage form |
The concentration of CFDA-SE is 10 μM |
|
Applications |
CFDA-SE, at 80 times the concentration used for in vitro labeling, was nontoxic and labeled randomly approximately 15% of thymocytes 24 h after injection. The turnover rate of labeled thymic emigrants in the lymph nodes was in the order of 21 days. Thus, CFDA-SE may serve as a powerful tool in relatively long-term migration studies. |
|
References: [1]: Wang XQ, Duan XM,et,al. Carboxyfluorescein diacetate succinimidyl ester fluorescent dye for cell labeling. Acta Biochim Biophys Sin (Shanghai). 2005 Jun;37(6):379-85. doi: 10.1111/j.1745-7270.2005.00051.x. PMID: 15944752. [2]: Graziano M, St-Pierre Y, et,al. The fate of thymocytes labeled in vivo with CFSE. Exp Cell Res. 1998 Apr 10;240(1):75-85. doi: 10.1006/excr.1997.3900. PMID: 9570923. |
|
| Cas No. | 150347-59-4 | SDF | |
| Überlieferungen | 5(6)Carboxyfluorescein diacetate succinimidyl ester, 5(6)-CFDA N-succinmidyl ester | ||
| Chemical Name | 5-(((2,5-dioxopyrrolidin-1-yl)oxy)carbonyl)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl diacetate | ||
| Canonical SMILES | O=C1N(OC(C2=CC=C(C3(C(C=CC(OC(C)=O)=C4)=C4OC5=C3C=CC(OC(C)=O)=C5)OC6=O)C6=C2)=O)C(CC1)=O | ||
| Formula | C29H19NO11 | M.Wt | 557.46 |
| Löslichkeit | ≥ 37.2mg/mL in DMSO with ultrasonic | Storage | Store at -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.7939 mL | 8.9693 mL | 17.9385 mL |
| 5 mM | 0.3588 mL | 1.7939 mL | 3.5877 mL |
| 10 mM | 0.1794 mL | 0.8969 mL | 1.7939 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 20 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
Required fields are marked with *