Editing Apoptosis (section)

==Methods for distinguishing apoptotic from necrotic cells==
[[File:Apoptosis in mouse pre-adipocytes.gif|thumb|Long-term live cell imaging (12h) of multinucleated mouse pre-Adipocyte trying to undergo mitosis. Due to the excess of genetic material the cell fails to replicate and dies by apoptosis.]]
Label-free [[live cell imaging]], [[time-lapse microscopy]], [[flow cytometry|flow fluorocytometry]], and [[transmission electron microscopy]] can be used to compare apoptotic and necrotic cells. There are  also various biochemical techniques for analysis of cell surface markers (phosphatidylserine exposure versus cell permeability by flow cytometry), cellular markers such as [[DNA fragmentation]]<ref>{{cite journal | vauthors = Lozano GM, Bejarano I, Espino J, González D, Ortiz A, García JF, Rodríguez AB, Pariente JA | year = 2009 | title = Density gradient capacitation is the most suitable method to improve fertilization and to reduce DNA fragmentation positive spermatozoa of infertile men | url = https://www.researchgate.net/publication/259983025 | journal = Anatolian Journal of Obstetrics & Gynecology | volume = 3 | issue = 1 | pages = 1–7 | access-date = 2016-03-08 | archive-date = 2022-04-30 | archive-url = https://web.archive.org/web/20220430005522/https://www.researchgate.net/publication/259983025_Density_gradient_capacitation_is_the_most_suitable_method_to_improve_fertilization_and_to_reduce_DNA_fragmentation_positive_spermatozoa_of_infertile_men | url-status = live }}</ref> (flow cytometry),<ref>{{cite journal | vauthors = Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F | title = Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis) | journal = Cytometry | volume = 27 | issue = 1 | pages = 1–20 | date = January 1997 | pmid = 9000580 | doi = 10.1002/(sici)1097-0320(19970101)27:1<1::aid-cyto2>3.0.co;2-l | doi-access =  }}</ref> caspase activation, Bid cleavage, and cytochrome c release ([[Western blot]]ting). Supernatant screening for caspases, HMGB1, and cytokeratin 18 release can identify primary from secondary necrotic cells. However, no distinct surface or biochemical markers of necrotic cell death have been identified yet, and only negative markers are available. These include absence of apoptotic markers (caspase activation, cytochrome c release, and oligonucleosomal DNA fragmentation) and differential kinetics of cell death markers (phosphatidylserine exposure and cell membrane permeabilization). A selection of techniques that can be used to distinguish apoptosis from necroptotic cells could be found in these references.<ref name="methods">{{cite book | vauthors = Krysko DV, Vanden Berghe T, Parthoens E, D'Herde K, Vandenabeele P | chapter = Chapter 16 Methods for Distinguishing Apoptotic from Necrotic Cells and Measuring Their Clearance | title = Programmed Cell Death, General Principles for Studying Cell Death, Part A | series = Methods in Enzymology | volume = 442 | pages = 307–41 | year = 2008 | pmid = 18662577 | doi = 10.1016/S0076-6879(08)01416-X | isbn = 9780123743121 | url = https://biblio.ugent.be/publication/431717/file/2936374 }}</ref><ref name="methods2">{{cite journal | vauthors = Krysko DV, Vanden Berghe T, D'Herde K, Vandenabeele P | title = Apoptosis and necrosis: detection, discrimination and phagocytosis | journal = Methods | volume = 44 | issue = 3 | pages = 205–221 | date = March 2008 | pmid = 18314051 | doi = 10.1016/j.ymeth.2007.12.001 }}</ref><ref name="methods3">{{cite journal | vauthors = Vanden Berghe T, Grootjans S, Goossens V, Dondelinger Y, Krysko DV, Takahashi N, Vandenabeele P | title = Determination of apoptotic and necrotic cell death in vitro and in vivo | journal = Methods | volume = 61 | issue = 2 | pages = 117–129 | date = June 2013 | pmid = 23473780 | doi = 10.1016/j.ymeth.2013.02.011 | url = https://zenodo.org/record/3423520 | access-date = 2019-11-05 | url-status = dead | archive-url = https://web.archive.org/web/20191105075341/https://zenodo.org/record/3423520 | archive-date = 2019-11-05 }}</ref><ref>{{cite book | vauthors = Wlodkowic D, Telford W, Skommer J, Darzynkiewicz Z | title = Recent Advances in Cytometry, Part B – Advances in Applications | chapter = Apoptosis and beyond: cytometry in studies of programmed cell death | series = Methods in Cell Biology | volume = 103 | pages = 55–98 | year = 2011 | pmid = 21722800 | pmc = 3263828 | doi = 10.1016/B978-0-12-385493-3.00004-8 | isbn = 9780123854933 }}</ref>