title: Fluorescein Diacetate Microplate Assay in Cell Viability Detection.
abstract: Objective To investigate the application of the fluorescein diacetate (FDA) microplate assay in cell viability detection. Methods Cells were seeded in a 96-well culture plate until detection. After incubated with FDA,the plate was detected by fluorescence microplate analyzer. The effects of FDA incubation duration,concentration,and other factors on the assay's accuracy and stability were assessed. We also compared the results of FDA with methyl thiazolyl(MTT) in terms of cell numbers and H2O2injury. Results Within 0-30 minutes,the fluorescence-cell number coefficient of FDA assay increased with duration and reached 0.99 in 27-30 minutes. The optimum concentration of final FDA in this study was 10-30 μg/ml. On cell viability detection,the result of FDA method was equivalent to MTT method. As to H2O2injury assay,the sensitivity of FDA method was superior to MTT on the higher concentration H2O2treatment due to a relative shorter duration for detection. Conclusion As a stable and reliable method,FDA is feasible for cell variability detection under varied conditions.
authors.affiliation: Beijing Key Laboratory of Drug Target Identification and Drug Screening,Institute of Materia Medica, CAMS and PUMC,Beijing 100050,China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Beijing Key Laboratory of Drug Target Identification and Drug Screening,Institute of Materia Medica, CAMS and PUMC,Beijing 100050,China.
authors.forename: Xiu-Ying
authors.initials: XY
authors.lastname: Yang
authors.affiliation: Beijing Key Laboratory of Drug Target Identification and Drug Screening,Institute of Materia Medica, CAMS and PUMC,Beijing 100050,China.
authors.forename: Lian-Hua
authors.initials: LH
authors.lastname: Fang
authors.affiliation: Beijing Key Laboratory of Drug Target Identification and Drug Screening,Institute of Materia Medica, CAMS and PUMC,Beijing 100050,China.
authors.forename: Guan-Hua
authors.initials: GH
authors.lastname: DU
mesh_terms: D001681:Biological Assay; D002470:Cell Survival; D005452:Fluoresceins / Q000737:chemistry*; D005453:Fluorescence; D006801:Humans; D006861:Hydrogen Peroxide; D013194:Staining and Labeling
chemical_list: D005452:Fluoresceins; D006861:Hydrogen Peroxide