Multimodal Holographic Microscopy: Distinction between Apoptosis and Oncosis

Balvan, Jan; Křížová, Aneta; Gumulec, Jaromír; Raudenská, Martina; Sládek, Zbyšek; Sedláčková, Miroslava; Babula, Petr; Svobodová, Markéta; Kizek, René; Chmelík, Radim; Masařík, Michal

Multimodal Holographic Microscopy: Distinction between Apoptosis and Oncosis

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journal.pone.0121674.PDF(2.23 MB)

Date

2015-03-24

Authors

Balvan, Jan

Křížová, Aneta

Gumulec, Jaromír

Raudenská, Martina

Sládek, Zbyšek

Sedláčková, Miroslava

Babula, Petr

Svobodová, Markéta

Kizek, René

Chmelík, Radim

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ORCID

Publisher

PLOS

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Abstract

Identification of specific cell death is of a great value for many scientists. Predominant types of cell death can be detected by flow-cytometry (FCM). Nevertheless, the absence of cellular morphology analysis leads to the misclassification of cell death type due to underestimated oncosis. However, the definition of the oncosis is important because of its potential reversibility. Therefore, FCM analysis of cell death using annexin V/propidium iodide assay was compared with holographic microscopy coupled with fluorescence detection Multimodal holographic microscopy (MHM). The aim was to highlight FCM limitations and to point out MHM advantages. It was shown that the annexin V+/PI phenotype is not specific of early apoptotic cells, as previously believed, and that morphological criteria have to be necessarily combined with annexin V/PI for the cell death type to be ascertained precisely. MHM makes it possible to distinguish oncosis clearly from apoptosis and to stratify the progression of oncosis.
Identifikace konkrétního druhu buněčné smrti je velmi významná pro mnoho vědců. Převládající druhy buněčné smrti mohou být detekovány metodou průtokové cytometrie (FCM). Nicméně, absence údajů o morfologii buněk vede ke špatnému určení druhu buněčné smrti kvůli podcenění onkózy. Proto do tohoto výzkumu byla zařazena multimodální holografická mikroskopie.

Keywords

Multimodal holographic microscopy, apoptosis, oncosis, necrosis, cell death, plumbagin, flow cytometry, transmission electron microscopy, Multimodální holografická mikroskopie, apoptóza, onkóza, nekróza, buněčná smrt, plumbagin, průtoková cytometrie, prozařovací elektronová mikroskopie

Citation

PLOS ONE. 2015, vol. 10, issue 3, p. 1-16.
https://journals.plos.org/plosone/article/authors?id=10.1371/journal.pone.0121674

Document type

Peer-reviewed

Document version

Published version

Language of document

en

Document licence

Creative Commons Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/