title: Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining.
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.
authors.affiliation: Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: sabine.pietkiewicz@med.ovgu.de.
authors.forename: Sabine
authors.initials: S
authors.lastname: Pietkiewicz
authors.affiliation: Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: joern.schmidt@med.ovgu.de.
authors.forename: Jörn H
authors.initials: JH
authors.lastname: Schmidt
authors.affiliation: Department of Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University, Magdeburg, Germany. Electronic address: inna.lavrik@med.ovgu.de.
authors.forename: Inna N
authors.initials: IN
authors.lastname: Lavrik
mesh_terms: D017304:Annexin A5 / Q000378:metabolism*; D017209:Apoptosis / Q000502:physiology*; D016923:Cell Death / Q000502:physiology; D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D019169:Jurkat Cells; D009336:Necrosis / Q000378:metabolism / Q000473:pathology*; D010718:Phosphatidylserines / Q000378:metabolism; D011419:Propidium / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D017304:Annexin A5; D010718:Phosphatidylserines; D011419:Propidium
keywords: Annexin V/propidium iodide; Apoptosis; Death receptor; Imaging flow cytometry; Necroptosis