Cell Viability Test,Cytoplasmic Streaming,Cell Mass,Formazan,Plasma Membrane,Fluorescein

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	Cell Viability Test - Cell viability can be determined by any one of the following approaches: (1) phase contrast microscopy, and staining with (2) 2, 3. 5-triphenyltetrazolium chloride (TIC), (3) fluorescein diacetate (FDA) and (4) Evan's blue. Live cells show cytoplasmic streaming and a well defined healthy nucleus which are easily observable with a phase contrast microscope or even a light microscope. Cell masses can be stained with 1-2% solution of TTC, which is reduced by living cells to formazan that yields red colour.
Formazan can be extracted and measured with a spectrophotometer to give a quantitative estimate of viability, but it is not suitable for single or few cells. Cell are treated with 0.01 % solution of FDA. Live cells cleave FDA by esterase activity and produce fluorescein, which can not cross plasma membrane. With UV exposure, fluorescein gives green fluorescence so that live cells appear green, while dead cells do not fluoresce. Evan's blue (0.025%) is not taken up by live cells, while it freely enters into damaged dead cells. Therefore all cells that take up stain are dead. Evan's blue is usually used in conjunction with FDA.