Flow Cytometry pp 355-362 | Cite as

Detection of Apoptosis using Fluorescent In Situ Nick Translation

  • R. R. Jonker
  • J. G. J. Bauman
  • J. W. M. Visser
Part of the NATO ASI Series book series (volume 67)


Apoptosis is the most common mechanism of cell death (Kerr et al., 1972). It is also known as Programmed Cell Death and it is dependent on specific enzymatic activity. Apoptosis takes place in embryogenesis, tumor regression, tissue atrophy and cell death induced by cytotoxic T-lymphocytes (Duvall and Wyllie, 1986). It can be induced by a number of factors, including glucocorticosteroids, tumor necrosis factor, lymphotoxin and radiation. The tumor suppressor gene bcl-2 blocks apoptosis as has been reviewed by Williams (1991). Wild-type p53 also functions as a tumor suppressor gene and induces apoptosis (Yonish-Rouach et al. 1991). Sellins and Cohen (1987) proved that, in contrast to necrosis, apoptosis is an active process by showing that induction of gene expression takes place after γ-irradiation. This gene expression is required for DNA fragmentation by endonucleases, an early event in apoptosis. Growth factors promote cell survival which means that they suppress apoptosis in haemopoietic cells (Williams et al., 1990, Nieto et al., 1989). Resting lymphocytes are very sensitive for irradiation and die via apoptosis. This process is known as Interphase Death. Sellins and Cohen (1987) suggested that this mechanism has a role in the elimination of cells that could be potentially harmful to the organism, because they might possibly turn into tumor cells by errors in the DNA repair process. Therefore they called this mechanism a ‘better dead than wrong’ mechanism that defends the organism against these potentially dangerous irradiated clonogenic lymphocytes.


Bone Marrow Cell Nick Translation FITC Fluorescence DAPI Fluorescence Flow Cytometric Detection 
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  1. Bayer JA, De Vries P, Herweijer H and Bauman JGJ (1990) The use of E. coli exonuclease III to generate single stranded DNA in BrdUrd cell-cycle analysis permits simultaneous detection of cell surface antigens. J. Immunol. Methods 132: 13–24.PubMedCrossRefGoogle Scholar
  2. Duke RC, Chervenak R and Cohen JJ (1983) Endogenous endonuclease-induced DNA fragmentation: An early event in cell mediated cytolysis. Proc. Natl. Acad. Sci. USA 80: 6361–6365.PubMedCrossRefGoogle Scholar
  3. Duvall E and Wyllie AH (1986) Death and the cell. Imm. Today 7: 115–119.CrossRefGoogle Scholar
  4. Kerem BS, Goitein R, Richler C, Marcus M, Cedar H (1983) In situ nick translation distinguishes between active and inactive X chromosomes. Nature 304, 88–90.PubMedCrossRefGoogle Scholar
  5. Kerr JFR, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics. Br. J. Cancer 26: 239.PubMedCrossRefGoogle Scholar
  6. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocytes apoptosis by propidium iodide staining and flow cytometry. J. Imm. Methods 139: 271–279.CrossRefGoogle Scholar
  7. Nieto MA, Lopez Rivas A (1989) IL-2 protects T lymphocytes from glucocorticoid-induced DNA fragmentation and cells death. J. of Immunology 143: 4166–4170.Google Scholar
  8. Schmid I, Krull WJ, Uittenbogaart CG, Braun J, Giorgi JV 1992. Dead cell discrimination with 7-Amino-Actinomycin D in combination with dual color immunofluorescence in single laser flow cytometry. Cytometry 13: 204–208.PubMedCrossRefGoogle Scholar
  9. Sellins KS, Cohen JJ (1987) Gene induction by γ-irradiation leads to DNA fragmentation in lymphocytes. J. of Immunology 139: 3199–3206.Google Scholar
  10. Swat W, Ignatowicz L, Kisielow P (1991) Detection of apoptosis of immature CD4+8+ thymocytes by flow cytometry. J. Imm. Methods 137: 79–87.CrossRefGoogle Scholar
  11. Williams GT, Smith CA, Spooncer E, Dexter TM, Taylor DR (1990) Haemopoietic colony stimulating factors promote cell survival by suppressing apoptosis. Nature 343: 76–79.PubMedCrossRefGoogle Scholar
  12. Williams GT (1991) Programmed cell death: Apoptosis and oncogenesis. Cell 65: 1097–1098.PubMedCrossRefGoogle Scholar
  13. Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, Oren, M (1991) Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6. Nature 352, 345–347.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • R. R. Jonker
    • 1
  • J. G. J. Bauman
    • 1
  • J. W. M. Visser
    • 1
  1. 1.TNO Institute of Applied Radiobiology and ImmunologyRijswijkThe Netherlands

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