Assessment of Cell Proliferation with Resazurin-Based Fluorescent Dye

  • Ewa M. CzekanskaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 740)


The Alamar Blue assay is based on enzymatic reduction of indicator dye by viable cells and serves as an effective tool for assessing cell proliferation and as a screening technique. It can be applied in studies concentrating on animal, plant, yeast, and bacteria cells. Among the various methods for cell viability and cytotoxicity, it utilises all features of ideal and reliable test; it is one-step, sensitive, safe, non-toxic for cells, and cost-effective.

Key words

Cell proliferation Resazurin Alamar Blue Metabolic activity Oxidoreductases 


  1. 1.
    Twigg R.S. (1945) Oxidation-reduction aspects of resazurin. Nature, 155:401–402CrossRefGoogle Scholar
  2. 2.
    Nixon M. C. and Lamb A. B. (1945) Resazurin Test for Grading Raw Milk. Can J Comp Med Vet Sci, 9(1):18–23PubMedGoogle Scholar
  3. 3.
    Ali-Vehmas T., Louhi M., and Sandholm M. (1991) Automation of the resazurin reduction test using fluorometry of microtitration trays. Zentralbl Veterinarmed B, 38(5):358–372PubMedGoogle Scholar
  4. 4.
    De Jong D. W. and. Woodlief W. G. (1977) Fluorimetric assay of tobacco leaf dehydrogenases with resazurin. Biochim Biophys Acta, 484(2):249–259PubMedGoogle Scholar
  5. 5.
    Glass R. H., Ericsson S. A., Ericsson R. J., Drouin M. T., Marcoux L. J., and Sullivan H. (1991) The resazurin reduction test provides an assessment of sperm activity. Fertil Steril, 56(4):743–746PubMedGoogle Scholar
  6. 6.
    To W. K., Fothergill A. W., and Rinaldi M. G. (1995) Comparative evaluation of macrodilution and alamar colorimetric microdilution broth methods for antifungal susceptibility testing of yeast isolates. J Clin Microbiol, 33(10):2660–2664PubMedGoogle Scholar
  7. 7.
    O’Brien J., Wilson I., Orton T., and Pognan F. (2000) Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem, 267(17):5421–5426PubMedCrossRefGoogle Scholar
  8. 8.
    Perrot S., Dutertre-Catella H., Martin C., Rat P., and Warnet J. M. (2003) Resazurin metabolism assay is a new sensitive alternative test in isolated pig cornea. Toxicol Sci, 72(1):122–129PubMedCrossRefGoogle Scholar
  9. 9.
    Hamid R., Rotshteyn Y., Rabadi L., Parikh R., and Bullock P. (2004) Comparison of alamar blue and MTT assays for high through-put screening. Toxicol In Vitro, 18(5):703–710PubMedCrossRefGoogle Scholar
  10. 10.
    Larson E. M., Doughman D. J., Gregerson D. S., and Obritsch W. F. (1997) A new, -simple, nonradioactive, nontoxic in vitro assay to monitor corneal endothelial cell viability. Invest Ophthalmol Vis Sci, 38(10):1929–1933PubMedGoogle Scholar
  11. 11.
    Jonsson K. B., Frost A., Larsson R., Ljunghall S., and Ljunggren O. (1997) A new fluorometric assay for determination of osteoblastic proliferation: effects of glucocorticoids and insulin-like growth factor-I. Calcif Tissue Int, 60(1):30–36PubMedCrossRefGoogle Scholar
  12. 12.
    O’Brien, J., Wilson, I., Orton, T., and Pognan, F. (2000) Pognan. Investigation of the Alamar blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem, 267, 5421–5426.Google Scholar
  13. 13.
    Gonzalez R. J. and Tarloff J. B. (2001) Evaluation of hepatic subcellular fractions for Alamar blue and MTT reductase activity. Toxicol In Vitro, 15(3):257–259PubMedCrossRefGoogle Scholar
  14. 14.
    Nakayama G. R., Caton M. C., Nova M. P., and Parandoosh Z. (1997) Assessment of the Alamar Blue assay for cellular growth and -viability in vitro. J Immunol Methods, 204(2):205–208PubMedCrossRefGoogle Scholar
  15. 15.
    De Jong, D. W. and Woodlief, W. G. (1977) Fluorometric assay of tobacco leaf dehydrogenases with resazurin. Biochim. Biophys. Acta, 484, 249–259.PubMedGoogle Scholar
  16. 16.
    Squatrito R. C., Connor J. P., and Buller R. E. (1995) Comparison of a novel redox dye cell growth assay to the ATP bioluminescence assay. Gynecol Oncol, 58(1):101–105PubMedCrossRefGoogle Scholar
  17. 17.
    Jonsson, K. B., Frost, A., Larsson, R., Ljunghall, S., and Ljunggren, O. (1997) A new fluorometric assay for determination of osteoblastic proliferation: effects of glucocorticoids and insulin-like growth factor-I. Calcif. Tissue Int, 60, 30–36.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.AO Research InstituteDavos PlatzSwitzerland

Personalised recommendations