Cell viability, fluorescein diacetate

Cell viability was assessed in all cell cultures remaining after the 48 hour time point was taken. Fluorescein diacetate was used to detect cell viability. Fluoreceine diacetate was added to the cell culture at a final concentration of 0.5% (w/v). Florescein diacetate is an ester which, as it enters a living cell, is... 

Determined on the basis of loss of cell membrane permeability in response to a deliberate modification in culture conditions. Cell viability can be determined by either neutral red or fluorescein diacetate retention assays. Volume 1(14,15). 

More recent staining procedures largely use fluorescent dyes to characterize the physiological and biochemical states of cells. Fluorescein Diacetate (FDA), a non-polar substance which crosses the membrane and is hydrolyzed by intracellular esterases in viable cells to produce fluorescein, exhibits yellow-green fluorescence when excited at 490 nm. Damaged or non-viable cells in general are unable to hydrolyze FDA or to retain fluorescein within the cell [172,173]. In combination with Ethidium Homodimer or Propidium Iodide, a similar esterase substrate, calcein acetoxy methyl ester (CAM) has been found to be reliable for viability assessment of protozoans, but not on Candida yeast, neither on bacteria such as Bacillus cereus and Escherichia coli [174]. 

Figure 2. Use of Fluorescein Diacetate to Estimate Cell Viability (See page I in color inserts,)...

Widholm, J.M. The Use of Fluorescein Diacetate and Phenosafranine for Determining Viability of Cultured Plant Cells, Stain Tech. 1972 47 189-194. 

Powell, H. M. Armour, A. D. Boyce, S. T. Fluorescein diacetate for determination of cell viability in 3D fibroblast-collagen-gag constructs. Methods Mol. Biol. 2011, 740,115-126. 

Nadel, B. L. The use of fluorescein diacetate as a viability probe for plant cells. Appl. Fluoresc. Technol. 1989,1,13-16. 

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