1. ¹INSERM, U848, F-94805 Villejuif, France
2. ²Institut Gustave Roussy, F-94805 Villejuif, France
3. ³Université Paris Sud-XI, F-94805 Villejuif, France
4. ⁴Department of Oncological Sciences, Mount Sinai School of Medicine, New York, NY 10029, USA
5. ⁵Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX 75390, USA
6. ⁶Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107-5587, USA
7. ⁷Department of Biochemistry and Biomedical Sciences, McMaster University, L8N 3Z5 Hamilton, Canada
8. ⁸Department of Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605-2324, USA
9. ⁹Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA
10. ¹⁰Roswell Park Cancer Institute, Buffalo, NY 14263, USA
11. ¹¹Center for Brain Repair and Rehabilitation, Institute of Neuroscience and Physiology, University of Gothenburg, SE-405 30 Gothenburg, Sweden
12. ¹²Department of Pediatric Oncology, The Queen Silvia Children's Hospital, SE-416 85 Gothenburg, Sweden
13. ¹³Institute of Molecular Medicine and Cell Research (ZBMZ), Albert-Ludwigs-Universität Freiburg, 79104 Freiburg, Germany
14. ¹⁴Buck Institute for Age Research, Novato, CA 94945, USA
15. ¹⁵University of California – San Francisco, San Francisco, CA 94143, USA
16. ¹⁶University of Versailles/St Quentin, 78035 Versailles, France
17. ¹⁷CNRS, UMR8159, 78035 Versailles, France
18. ¹⁸National Institutes of Environmental Health Sciences, NIH, Duhram, NC 27709, USA
19. ¹⁹Vascular and Tumor Biology Research Center, The Rappaport Faculty of Medicine, Technion – Israel Institute of Technology, 31096 Haifa, Israel
20. ²⁰Medical Research Council, Toxicology Unit, Leicester University, Leicester LE1 9HN, UK
21. ²¹Dipartimento di Scienze Biomediche, Università 'G. d'Annunzio' Chieti-Pescara, 66100 Chieti, Italy
22. ²²Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, 00161 Rome, Italy
23. ²³Mediterranean Institute of Oncology, 95030 Catania, Italy
24. ²⁴Neuroscience Center, Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599-7250, USA
25. ²⁵Department of Pathology, New York University School of Medicine, New York, NY 10016, USA
26. ²⁶Hematology-Oncology Division, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA
27. ²⁷Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA
28. ²⁸Howard Hughes Medical Institute, Chevy Chase, MD 20815-6789, USA
29. ²⁹University Children's Hospital, 89075 Ulm, Germany
30. ³⁰INSERM, UMR866, 21049 Dijon, France
31. ³¹Faculty of Medicine and Pharmacy, University of Burgundy, 21049 Dijon, France
32. ³²INSERM, U631, 13288 Marseille, France
33. ³³CNRS, UMR6102, 13288 Marseille, France
34. ³⁴Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, 13288 Marseille, France
35. ³⁵Institut Pasteur, Antiviral Immunity, Biotherapy and Vaccine Unit, 75015 Paris, France
36. ³⁶Department of Immunology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA
37. ³⁷Department of Cancer Biology – Institut de Génétique et de Biologie Moléculaire et Cellulaire, 67404 Illkirch, France
38. ³⁸CNRS, UMR7104, 67404 Illkirch, France
39. ³⁹INSERM, U964, 67404 Illkirch, France
40. ⁴⁰Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA
41. ⁴¹Department of Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, MD 21205, USA
42. ⁴²Institute of Molecular Biology, University of Zurich, 8057 Zurich, Switzerland
43. ⁴³Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan
44. ⁴⁴Danish Cancer Society, Department of Apoptosis, Institute of Cancer Biology, DK-2100 Copenhagen, Denmark
45. ⁴⁵Department of Molecular Genetics, Weizmann Institute of Science, 76100 Rehovot, Israel
46. ⁴⁶Life Sciences Institute and Department of Molecular, Cellular, and Developmental Biology and Biological Chemistry, University of Michigan, Ann Arbor, MI 48109, USA
47. ⁴⁷Institute of Child Health, University College London, London WC1N 1EH, UK
48. ⁴⁸Duke University School of Medicine, Durham, NC 27710, USA
49. ⁴⁹Centre for Cancer Biology, Hanson Institute, Adelaide, South Australia 5000, Australia
50. ⁵⁰Southwestern Medical Center, University of Texas, Dallas, TX 75390, USA
51. ⁵¹Burnham Institute for Medical Research, La Jolla, CA 92037, USA
52. ⁵²The Salk Institute for Biological Studies, La Jolla, CA 92037, USA
53. ⁵³The Scripps Research Institute, La Jolla, CA 92037, USA
54. ⁵⁴Univerisity of California-San Diego, La Jolla, CA 92093, USA
55. ⁵⁵Immunotechnology Section, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, USA
56. ⁵⁶Institute of Molecular Biosciences, University of Graz, 8010 Graz, Austria
57. ⁵⁷Department of Therapeutic Research and Medicines Evaluation, Section of Cell Aging and Degeneration, Istituto Superiore di Sanità, 00161 Rome, Italy
58. ⁵⁸Laboratory for Molecular Cancer Biology, VIB, 9052 Ghent, Belgium
59. ⁵⁹Department for Molecular Biology, Ghent University, 9052 Ghent, Belgium
60. ⁶⁰Department of Genetics, Trinity College, Dublin 2, Ireland
61. ⁶¹Center for Experimental and Molecular Medicine, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
62. ⁶²University of Amsterdam, 1012 ZA Amsterdam, The Netherlands
63. ⁶³Apoptosis, Cancer, and Development Laboratory, Centre Léon Berard, 69008 Lyon, France
64. ⁶⁴CNRS, UMR5238, 69008 Lyon, France
65. ⁶⁵Université de Lyon, 69008 Lyon, France
66. ⁶⁶Department of Experimental Medicine and Biochemical Sciences, University of Rome 'Tor Vergata', 00133 Rome, Italy
67. ⁶⁷Department of Pathology, Stony Brook University, Stony Brook, NY 11794-8691, USA
68. ⁶⁸Department of Molecular Oncology, Göttingen Center of Molecular Biosciences, 37077 Göttingen, Germany
69. ⁶⁹Faculty of Medicine, University of Göttingen, 37077 Göttingen, Germany
70. ⁷⁰Department of Medical Chemistry, Graduate School of Medicine, University of Kyoto, Kyoto 606-8501, Japan
71. ⁷¹Merck Research Laboratories, Rahway, NJ 07065-0900, USA
72. ⁷²University of Michigan Medical School, Ann Arbor, MI 48109, USA
73. ⁷³Department of Molecular Cell Biology, Weizmann Institute of Science, 76100 Rehovot, Israel
74. ⁷⁴Institute of Molecular Biotechnology of the Austrian Academy of Science, 1030 Vienna, Austria
75. ⁷⁵Department of Physiology, Yong Loo Lin School of Medicine, Graduate School for Integrative Sciences and Engineering, National University of Singapore, 117597 Singapore
76. ⁷⁶Singapore-MIT Alliance, National University of Singapore, 117576 Singapore
77. ⁷⁷Duke-NUS Graduate Medical School, 169547 Singapore
78. ⁷⁸Ben May Department for Cancer Research, University of Chicago, Chicago, IL 60637, USA
79. ⁷⁹Laboratory of Cell Biology, National Institute for Infectious Diseases IRCCS 'L. Spallanzani', 00149 Rome, Italy
80. ⁸⁰Department of Biology, University of Rome 'Tor Vergata', 00133 Rome, Italy
81. ⁸¹Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, Dublin 2, Ireland
82. ⁸²Department of Biochemistry, La Trobe University, 3086 Victoria, Australia
83. ⁸³Laboratorio de Inmunopatología, Instituto de Biología y Medicina Experimental (IBYME- CONICET), C1428 Buenos Aires, Argentina
84. ⁸⁴Department of Biomedical Sciences, University of Padova, 35121 Padova, Italy
85. ⁸⁵iMed.UL, Faculty of Pharmacy, University of Lisbon, 1649-003 Lisbon, Portugal
86. ⁸⁶Cambridge Institute for Medical Research, Cambridge CB2 0XY, UK
87. ⁸⁷Biocenter, University of Würzburg, 97074 Würzburg, Germany
88. ⁸⁸Department of Cell Physiology and Metabolism, University of Geneva Medical School, 1211 Geneva, Switzerland
89. ⁸⁹Dulbecco-Telethon Institute, Venetian Institute of Molecular Medicine, 35129 Padova, Italy
90. ⁹⁰Department of Pharmacology, University of Bern, 3010 Bern, Switzerland
91. ⁹¹Laboratory of Apoptosis and Cancer Biology, The Rockefeller University, New York, NY 10065, USA
92. ⁹²Department of Biochemistry, University of Lausanne, 1066 Epalinges, Switzerland
93. ⁹³Department of Medical Genetics, Osaka University Medical School, Osaka 565-0871, Japan
94. ⁹⁴Department for Molecular Biomedical Research, VIB, 9052 Ghent, Belgium
95. ⁹⁵The Beatson Institute for Cancer Research, Glasgow G61 1BD, UK
96. ⁹⁶Biochemistry Section, Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD 20892, USA
97. ⁹⁷Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA
98. ⁹⁸Institute of Environmental Medicine, Division of Toxicology, Karolinska Institute, SE- 171 77 Stockholm, Sweden

Correspondence: G Kroemer, INSERM, U848, Institut Gustave Roussy, PR1, 39, rue Camille Desmoulins, F-94805 Villejuif, France. Tel: +33-1-4211-6046; Fax: +33-1-4211-6047; E-mail: kroemer@orange.fr

Received 12 March 2009; Accepted 17 March 2009; Published online 17 April 2009.

Abstract

Cell death is essential for a plethora of physiological processes, and its deregulation characterizes numerous human diseases. Thus, the in-depth investigation of cell death and its mechanisms constitutes a formidable challenge for fundamental and applied biomedical research, and has tremendous implications for the development of novel therapeutic strategies. It is, therefore, of utmost importance to standardize the experimental procedures that identify dying and dead cells in cell cultures and/or in tissues, from model organisms and/or humans, in healthy and/or pathological scenarios. Thus far, dozens of methods have been proposed to quantify cell death-related parameters. However, no guidelines exist regarding their use and interpretation, and nobody has thoroughly annotated the experimental settings for which each of these techniques is most appropriate. Here, we provide a nonexhaustive comparison of methods to detect cell death with apoptotic or nonapoptotic morphologies, their advantages and pitfalls. These guidelines are intended for investigators who study cell death, as well as for reviewers who need to constructively critique scientific reports that deal with cellular demise. Given the difficulties in determining the exact number of cells that have passed the point-of-no-return of the signaling cascades leading to cell death, we emphasize the importance of performing multiple, methodologically unrelated assays to quantify dying and dead cells.