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ISSN 1672-9145 Acta Biochim Biophys Sin 2005, 37(6): 379–385 CN 31-1940/Q

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Carboxyfluorescein Diacetate Succinimidyl Ester Fluorescent Dye for Cell Labeling

Xiao-Qi WANG, Xiu-Mei DUAN, Li-Hua LIU, Yan-Qiu FANG, and Yan TAN*

The Central Laboratory, the First Teaching Hospital of Jilin University, Changchun 130021, China

Abstract Our objective was to study the properties of the carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and the methodology of cell labeling using CFDA-SE fluorescent dye. First, we analyzed the kinetics of CFDA-SE fluorescent dye intensity over time. Second, we determined the optimal concentration of CFDA-SE fluorescent dye for cell labeling. Third, we tested the toxicity of CFDA-SE fluorescent dye on labeled cells. Finally, we determined the optimal staining time of CFDA-SE fluorescent dye for cell labeling. The results show that the optimal concentration of CFDA-SE fluorescent dye for cell labeling varies according to different cell types. CFDA-SE fluorescent dye is non-toxic to cells as the cell death rate caused by CFDA-SE labeling is below 5%. The optimal cell labeling time was determined to be 8 min of incubation with CFDA-SE fluorescent dye. We concluded that the advantages of using CFDA-SE fluorescent dye for cell labeling are as follows: (1) the binding of CFDA-SE fluorescent dye to cells is stable; (2) CFDA-SE fluorescent dye is not toxic and does not modify the viability of labeled cells; and (3) CFDA-SE fluorescent dye is a suitable fluorochrome for cell labeling.

Key words carboxyfluorescein diacetate succinimidyl ester (CFDA-SE); carboxyfluorescein succinimidyl ester (CFSE); flow cytometry

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Received: December 13, 2004 Accepted: April 7, 2005

This work was supported by the grants from the National Education Ministry, State Key Basic Research Program of China (No. 20013145), the Key Technological Program of the Government Agency of Science and Technology of Jilin Province (No. 20020403, No. 20040414), the Scientific and Technological Program of Developing Industry Base of Changchun (04-02GG218)

*Corresponding author: Tel, 86-431-5668196; E-mail, [email protected]