Measuring apoptosis by microscopy and flow cytometry

Emilie Hollville, Seamus J. Martin

Research output: Contribution to journalArticlepeer-review

25 Citations (Scopus)

Abstract

Apoptosis is a mode of programmed cell death that plays an important role during development and in the maintenance of tissue homeostasis. Numerous physiological as well as pathological stimuli trigger apoptosis such as engagement of Fas, TRAIL, or TNF receptors, growth factor deprivation, hypoxia, or exposure to cytotoxic drugs. Apoptosis is coordinated from within by members of the caspase family of cysteine proteases that, upon activation, trigger a series of morphological changes including cell shrinkage, extensive plasma membrane blebbing, chromatin condensation, DNA hydrolysis, and nuclear fragmentation. These dramatic structural and biochemical alterations result not only in the controlled dismantling of the cell, but also in the efficient recognition and removal of apoptotic cells by phagocytes. Necrosis, which is typically nonprogrammed or imposed upon the cell by overwhelming membrane or organelle damage, is characterized by rapid plasma membrane rupture followed by organelle and cell swelling. Necrosis is often provoked by infectious agents or a severe departure from physiological conditions. This unit describes protocols for the measurement of apoptosis and for distinguishing apoptosis from necrosis.

Original languageEnglish
Pages (from-to)14.38.1-14.38.24
JournalCurrent Protocols in Immunology
Volume2016
DOIs
Publication statusPublished - 2 Feb 2016

Bibliographical note

Funding Information:
The Martin laboratory is supported by PI (14/IA/2622) and SRC (07/SRC/B1144) grants from Science Foundation Ireland. SJM is a Science Foundation Ireland Principal Investigator.

Publisher Copyright:
© 2016 by John Wiley & Sons, Inc.

Keywords

  • Apoptosis
  • Cell morphology
  • Flow cytometry
  • Microscopy
  • Necrosis

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