Detection of Apoptosis by TUNEL Assay

  • Kateryna Kyrylkova
  • Sergiy Kyryachenko
  • Mark Leid
  • Chrissa KioussiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 887)


Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay has been designed to detect apoptotic cells that undergo extensive DNA degradation during the late stages of apoptosis. The method is based on the ability of TdT to label blunt ends of double-stranded DNA breaks independent of a template. This chapter describes an assay for detection of apoptotic cells during mouse odontogenesis using a colorimetric TUNEL system.

Key words

Apoptosis TUNEL Immunofluorescence Mouse embryos Frozen sections 


  1. 1.
    Saraste A. (1999) Morphologic criteria and detection of apoptosis. Herz 24 (3), 189–195.PubMedCrossRefGoogle Scholar
  2. 2.
    Saraste A., Pulkki K. (2000) Morphologic and biochemical hallmarks of apoptosis. Cardiovasc Res 45 (3), 528–537.PubMedCrossRefGoogle Scholar
  3. 3.
    Cohen J. J. (1993) Apoptosis. Immunol Today 14 (3), 126–130.PubMedCrossRefGoogle Scholar
  4. 4.
    Bortner C. D., Oldenburg N. B., Cidlowski J. A. (1995) The role of DNA fragmentation in apoptosis. Trends Cell Biol 5 (1), 21–26.PubMedCrossRefGoogle Scholar
  5. 5.
    Gavrieli Y., Sherman Y., Ben-Sasson S. A. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119 (3), 493–501.PubMedCrossRefGoogle Scholar
  6. 6.
    Nagoescu A., Lorimier P., Labat-Moleur F., Drouet C., Robert C., Guillermet C., Brambilla C., Brambilla E. (1996) In situ apoptotic cell labeling by the TUNEL method: improvement and evaluation on cell preparations. J Histochem Cytochem 44 (9), 959–968.CrossRefGoogle Scholar
  7. 7.
    Rode H. D., Eisel D., Frost I. (2004) Apoptosis, cell death and cell proliferation. 3rd ed. London: Roche Applied Science.Google Scholar
  8. 8.
    Grasl-Kraupp B., Ruttkay-Nedecky B., Koudelka H. Bukowska K., Bursch W., Schulte-Hermann R. (1995) In situ detection of fragmented DNA (TUNEL assay) fails to discriminate among apoptosis, necrosis, and autolytic cell death: a cautionary note. Hepatology 21 (5), 1465–1468.PubMedGoogle Scholar
  9. 9.
    Orita Y., Nishizaki K., Sasaki J., Kanda S., Kimura N., Nomiya S., Yuen K., Masuda Y. (1999) Does TUNEL staining during peri- and post-natal development of the mouse inner ear indicate apoptosis? Acta Otolaryngol Suppl 540, 22–26.PubMedGoogle Scholar
  10. 10.
    Tornusciolo D. R., Schmidt R. E., Roth K. A. (1995) Simultaneous detection of TDT-mediated dUTP-biotin nick end-labeling (TUNEL)-positive cells and multiple immunohistochemical markers in single tissue ­sections. Biotechniques 19 (5), 800–805.PubMedGoogle Scholar
  11. 11.
    Oberhaus S. M. (2003) TUNEL and immunofluorescence double-labeling assay for apoptotic cells with specific antigen(s). Methods Mol Biol 218, 85–96.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Kateryna Kyrylkova
    • 1
  • Sergiy Kyryachenko
    • 1
  • Mark Leid
    • 1
  • Chrissa Kioussi
    • 2
    Email author
  1. 1.Department of Pharmaceutical Sciences, College of PharmacyOregon State UniversityCorvallisUSA
  2. 2.College of Pharmacy, Department of Pharmaceutical SciencesOregon State UniversityCorvallisUSA

Personalised recommendations