| Description | CFSE (CFDA-SE) is a fluorescent dye with cell membrane permeability. CFSE irreversibly binds to intracellular proteins in living cells and is used for the detection of cell proliferation. The labeled cells fluoresce in green color with excitation wavelength of 488 nm and emission wavelength of 518 nm. |
| In vitro | 方法：将 1 mL 细胞和 CFDA (5 mM in 110μL PBS) 翻转混匀以标记细胞。CFSE 标记的 CD8+OT-I T 细胞与用不同量的 OVA 脉冲的树突细胞培养 3 天，使用 Flow Cytometry 检测 CFSE 图谱。结果：CD8+ T 细胞根据 CFSE 稀释峰分裂 1-3 次，更多的 T 细胞在较高的抗原浓度下分裂。[1]方法：人红白血病细胞系K562、小鼠淋巴瘤细胞系YAC-1、人乳腺癌症细胞 MCF-7 和 人黑色素瘤细胞系 A375 用 CFDA (1-10 mM) 处理 1-6 h，使用 Flow Cytometry 检测细胞死亡情况。结果：CFDA 对细胞无毒，因为 CFDA 标记引起的细胞死亡率低于 5%。[2] |
| In vivo | 方法：CFSE 标记的 CD8+OT-I T 细胞尾静脉注射给 C56BL/6J 小鼠，随后静脉注射 OVA (20 μg)，三天后检测 CFSE 图谱。结果：大多数细胞落在 7 个 CFSE 峰值内，这表明细胞已经经历了多达 6 次分裂。[1]方法：为在体内标记胸腺细胞，将 CFDA (10 μM) 注射到麻醉 C56BL/6 小鼠的胸腺叶。结果：CFDA 标记了所有胸腺细胞亚群的代表性样本，并且这些细胞以大约 2-3×10^6 个细胞/天的速率迁移到外周淋巴器官。它们在注射后第 1 天进入淋巴结，并停留至少 21 天，而在脾脏中的周转更快。[3] |
| Cell experiments | The 5 mM CFDA-SE stock in DMSO is 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. After each cell line is harvested and washed three times with PBS, 1×109 cells are 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. The labeling reaction is stopped for 1 min by adding an equal volume of heat inactivated fetal bovine serum. The CFDA-SE labeled cells are washed twice with PBS and recounted, and the cell concentration is adjusted to 6×104cells/ml in IMDM containing 10% FCS.(Only for Reference) |
| Synonyms | 5(6)-CFDA N-succinmidyl ester, Carboxyfluorescein succinimidyl ester, CFDA-SE, 5(6)-羧基二乙酸荧光素琥珀酰亚胺酯, 5(6)-Carboxyfluorescein diacetate succinimidyl ester |
| molecular weight | 557.46 |
| Molecular formula | C29H19NO11 |
| CAS | 150347-59-4 |
| Storage | keep away from direct sunlight | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| Solubility | Ethanol: < 1 mg/mL (insoluble or slightly soluble) DMSO: 93 mg/mL (166.8 mM) H2O: < 1 mg/mL (insoluble or slightly soluble) |
| References | 1. Quah BJ, et al. The use of carboxyfluorescein diacetate succinimidyl ester (CFSE) to monitor lymphocyte proliferation. J Vis Exp. 2010 Oct 12;(44):2259. 2. Wang XQ, et al. Carboxyfluorescein diacetate succinimidyl ester fluorescent dye for cell labeling. Acta Biochim Biophys Sin (Shanghai). 2005 Jun;37(6):379-85. 3. Graziano M, et al. The fate of thymocytes labeled in vivo with CFSE. Exp Cell Res. 1998 Apr 10;240(1):75-85. 4. Li M, Qin M, Song G, et al. A biomimetic antitumor nanovaccine based on biocompatible calcium pyrophosphate and tumor cell membrane antigens[J]. Asian Journal of Pharmaceutical Sciences. 2020 |
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