Dojindo/CFSE/10/C309,CFSE 标记细胞

CFSE 是细胞膜可渗透的，并且很容易在活细胞内积聚，在那里它与细胞内蛋白质共价结合（图 1）。 水解的 CFSE 发出荧光，共价连接的荧光素分子不会从细胞中泄漏。 CFSE 标记的细胞可以在体内监测数周。 因此，CFSE 可用于检测活细胞以及通过荧光显微镜对细胞活动进行长期观察。 CFSE 标记细胞的激发和发射波长分别为 500 nm 和 520 nm。

Staining Procedure1. Prepare 1 mM CFSE solution with DMSO. Dilute it to prepare 10-50 μM CFSE solution with PBS or an appropriate buffer.2. Add CFSE solution with 1/10 of the volume of cell culture medium to the cell culture.3. Incubate the cell at 37ºC for 15 to 30 min.4. Wash cells twice with PBS or an appropriate buffer.5. Observe the cells under a fluorescence microscope with 490 nm excitation and 530 nm emission filters.

1. M. Bronner-Fraser, Alterations in Neural Crest Migration by a Monoclonal Antibody That Affects Cell Adhesion. J Cell Biol. 1985;101:610-617.2. A. Nose, et al., A Novel Cadherin Cell Adhesion Molecule:Its Expression Patterns Associated WithImplantation and Organogenesis of Mouse Embryos. J Cell Biol. 1986;103:2649-2658.3. S. A. Weston, et al., New Fluorescent Dyes for Lymphocyte Migration Studies Analysis by Flow Cytometry and Fluorescent Microscopy. J Immunol Methods. 1990;133:87-97.4. C. K. Raymond, et al., Molecular Analysis of the Yeast VPS3 Gene and the Role of Its Product in Vacuolar Protein Sorting and Vacuolar Segregation during the Cell Cycle. J Cell Biol. 1990;111:877-892.5. G. Radcliff, et al., Quantification of Effector/Target Conjugation Involving Natural Killer(NK) or Lymphokine Activated Killer(LAK) Cells by Two-color Flow Cytometry. J Immunol Methods. 1991;139:281-292.6. S. A. Weston, et al., Calcein: a Novel Marker for Lymphocytes Which Enter Lymph Nodes. Cytometry. 1992;13:739-749.7. L. S. D. Clerck, et al., Use of Fluorescent Dyes in the Determination of Adherence of Human Leucocytes to Endothelial Cells and the Effects of Fluorochromes on Cellular Function. J Immunol Methods. 1994;172:115-124.