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J Immunol Methods. 2015 Aug;423:99-103. doi: 10.1016/j.jim.2015.04.025. Epub 2015 May 11.

Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining.

Author information

1
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: sabine.pietkiewicz@med.ovgu.de.
2
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: joern.schmidt@med.ovgu.de.
3
Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: inna.lavrik@med.ovgu.de.

Abstract

Precisely identifying the type of programmed cell death is one of the key questions in contemporary biomedical research. We developed a straightforward approach allowing quantitative discrimination between two types of cell death on the single cell level: apoptosis and necroptosis. This method uses the combination of imaging flow cytometry with classical Annexin V/propidium iodide staining, which allows for the ascertainment of typical features of dying cells: exposure of the phospholipid phosphatidylserine and the loss of membrane integrity. Image-based analysis of nuclear morphology enables us to distinguish between secondary necrotic/late apoptotic and necroptotic cells directly in one assay. This is a major advantage compared to other contemporary approaches of necroptosis detection, which require a parallel application of several methods. This approach can be used for the quantitative assessment of cell death in cell and systems biology studies of signal transduction networks.

KEYWORDS:

Annexin V/propidium iodide; Apoptosis; Death receptor; Imaging flow cytometry; Necroptosis

PMID:
25975759
DOI:
10.1016/j.jim.2015.04.025
[Indexed for MEDLINE]

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