Methods to detect apoptotic cell death

Abstract

In concert with the increased understanding that there are many ways for cells to die, several methods have been developed to detect cell death. The classification of cell death posed some difficulties that were overcome by implementing strict selection criteria that should also apply to the detection methods. The selection of assays is based on morphological criteria and distinguishable marks of apoptotic patways. The detection of apoptosis includes methods related to membrane alterations, DNA fragmentation, cytotoxicity and cell proliferation, mitochondrial damage, immunological detection and mechanism based assays. Other less frequently used detections of apoptosis are: (a) light-scattering flow cytometry to avoid underestimating the extent and timing of apoptosis, (b) time-lapse microscopy perfusion platform to support the temporal aspects of detection, to measure cell surface area and cellular adhesion, and (c) genotoxicity specific chromatin changes. Attention is called to the advantages and limitations of various methods.

The immune response to secondary necrotic cells

“…The 2009 published recommendations of the Nomenclature Committee on Cell Death (NCCD) defined biochemical analyses that are necessary for an accurate identification of apoptotic cell death (e.g. TUNEL assay, active caspase 3) [ 100 , 101 ]. The authors note that there is no common biochemical denominator for necrotic cell death which can be used in tissue sections.…”

Comparison of cytological and immunocytochemical methods for detecting apoptotic epithelial cells in cervicovaginal smears

“…We believe that we need cost and time efficient methods to detect/evaluate apoptosis during the examination of cytological smears. Various cytological methodologies have been defined in order to detect and evaluate apoptosis . In this study, we aimed to evaluate whether Pap‐stained smears were reliable or not in the detection of apoptosis and to establish a new cytological scoring model called “Hacettepe Apoptosis Cytology Scoring (HA‐C score) system” for the cytological evaluation of apoptosis.…”

Spontaneous and activation-induced apoptosis of peripheral blood mononuclear cells in the pathogenesis of type 1 diabetes mellitus

“…Апоптоз лимфоцитов и диабет 1 типа Lymphocyte apoptosis and diabetes 2020, Vol. 22 [4,6,12]. В основе метода лежит взаимодействие красителя этидиум бромида с ДНК апоптотических клеток, после чего исследуемые образцы подвергают проточной цитометрии.…”

“…В основе метода лежит взаимодействие красителя этидиум бромида с ДНК апоптотических клеток, после чего исследуемые образцы подвергают проточной цитометрии. Процент клеток во фракции, соответствующей гиподиплоидному содержанию ДНК, характеризует апоптотические процессы [12]. Фракция апоптотических клеток (М1) располагается левее основного пика (М2), соответствующего диплоидным клеткам на гистограмме (рис.…”