Product DescriptionFDA is cell-membrane permeable and accumulates inside of viable cells as fluorescein (Fig. 1). Since fluorescein is less hydrophilic than BCECF or Calcein, the leakage of fluorescein from cells is rather high. FDA is also utilized for flow cytometry. The excitation and emission wavelengths of fluorescein are 488 nm and 530 nm, respectively. FDA-stained cells are shown in Fig. 2.
Fig. 1 Cell staining mechanism
Staining Procedure1.Prepare 0.5 mg/ml FDA stock solution with DMSO. Dilute 10 ul of the stock solution with 5 ml PBS(-).2.Prepare a cell suspension and wash cells with PBS(-). Prepare 1×105-1×106 cells/ml cell suspension3.Add 15 ul FDA solution to 30 ul cell suspension, and incubate at 37ºC for 15-30 min.4.Put 10 ul stained cell suspension on a glass slide and cover with a cover glass.5.Observe the cells under a fluorescence microscope with 488 nm excitation and 530 nm emission filters.
1. B. Rotman, et al., Membrane Properties of Living Mammalian Cells as Studied by Enzymatic Hydrolysis of Fluorogenic Esters. PNAS. 1966;55:134-141.2. H. R. Hulett, et al., Cell Sorting: Automated Separation of Mammalian Cells as a Function of Intercellular Fluorescence. Science. 1969;166:747-749.3. K. H. Jones, et al., An Improved Method to Determine Cell Viability by Simultaneous Staining with Fluorescein Diacetate-Propidium Iodide. J Histochem Cytochem. 1985;33:77-79.4. K. McGinnes, et al., A Fluorescence NK Assay Using Flow Cytometry. J Immunol Methods. 1986;86:7-15.5. W. M. J. Vuist, et al., Potentiation by Interleukin 2 of Burkitt’s Lymphoma Therapy with Anti-Pan B (Anti-CD19) Monoclonal Antibodies in a Mouse Xenotransplantation Model. Cancer Res. 1989;49:3783-3788.6. E. Prosperi, Intracellular Turnover of Fluorescein Diacetate. Influence of Membrane Ionic Gradients on Fluorescein Efflux. Histochem J. 1990;22:227-233.
Fig. 2 Cell staining with FDA,Cell type: HeLa
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