An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide

J Histochem Cytochem. 1985 Jan;33(1):77-9. doi: 10.1177/33.1.2578146.


A rapid, simultaneous double-staining procedure using fluorescein diacetate (FDA) and propidium iodide (PI) is described for use in the determination of cell viability in cell suspension. Air-dried slide preparations can be made from the cell suspensions so that an accurate estimate of the viability of the cells in the original suspension can be made up to 1 week later. Viable cells fluoresce bright green, while nonviable cells are bright red. Furthermore, when FDA-PI staining is compared to trypan blue dye exclusion as a method to determine cell viability, FDA-PI is found to be more consistent over prolonged periods of exposure to the dyes. Therefore, double staining with FDA-PI is a rapid, convenient, and reliable method to determine cell viability.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Count
  • Cell Survival*
  • Fluoresceins*
  • Histocytochemistry / methods
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence / methods
  • Phenanthridines*
  • Propidium*
  • Spleen / cytology
  • Staining and Labeling / methods*
  • Trypan Blue


  • Fluoresceins
  • Phenanthridines
  • Propidium
  • Trypan Blue
  • diacetylfluorescein