Record Display for the EPA National Library Catalog


Main Title Limitations of the Fluorescent Probe Viability Assay.
Author Massaro, E. J. ; Elstein, K. H. ; Bair, K. W. ; Zucker, R. M. ;
CORP Author Health Effects Research Lab., Research Triangle Park, NC. ;ManTech Environmental Technology, Inc., Research Triangle Park, NC.
Publisher c1989
Year Published 1989
Report Number EPA/600/J-89/551;
Stock Number PB92-113166
Additional Subjects Cell survival ; Fluorescent dyes ; Toxic substances ; Flow cytometry ; Tributyltin ; Acute erythroblastic leukemia ; Propidium ; Antineoplastic agents ; Cell division ; Growth ; Reprints ;
Library Call Number Additional Info Location Last
NTIS  PB92-113166 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 16p
Cell viability commonly is determined flow cytometrically by the carboxyfluorescein diacetate (CFDA)/propidium iodide (PI) assay. CFDA is taken up by the viable cell and converted via cytoplasmic esterase-catalyzed hydrolysis to carboxyfluorescein (CF). CF fluorescence intensity is considered to be an index of cellular vigor. It is generally accepted that the viable cell excludes PI. PI uptake is indicative of irreversible cellular injury and presages cell death. The authors observe that, following incubation for 4 hr with 0.5 - 1.0 microMolar tributyltin (TBT), murine erythroleukemic cells (MELC) exhibit supranormal CF fluorescence and exclude PI. Apparent cell volume is unaltered. However, the rate of growth (cell duplication) of these cells is depressed, suggesting that supranormal CF fluorescence, even in the absence of PI uptake, is indicative of cellular perturbation. Furthermore, at higher TBT concentrations (>1.0, <50.0 microMolar), the cells exhibit both increased CF fluorescence and PI fluorescence and are growth inhibited. These observations indicate that, by and of itself, CF fluorescence is neither a reliable indicator of cell viability nor vigor and suggest that at least in the case of perturbed cells, viability/growth assays based on intrinsic enzyme activities potentially are unreliable and inaccurate.