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Multimodal holographic microscopy: distinction between apoptosis and oncosis

J. Balvan, A. Krizova, J. Gumulec, M. Raudenska, Z. Sladek, M. Sedlackova, P. Babula, M. Sztalmachova, R. Kizek, R. Chmelik, M. Masarik,

. 2015 ; 10 (3) : e0121674. [pub] 20150324

Language English Country United States

Document type Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc16010416

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.

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$a Balvan, Jan $u Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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$a 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.
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$a Krizova, Aneta $u Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic; TESCAN Brno, s.r.o., Brno, Czech Republic.
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$a Gumulec, Jaromir $u Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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$a Raudenska, Martina $u Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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$a Sladek, Zbysek $u Department of Morphology, Physiology, and Animal Genetics, Mendel University in Brno, Brno, Czech Republic.
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$a Sedlackova, Miroslava $u Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
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$a Babula, Petr $u Department of Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
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$a Sztalmachova, Marketa $u Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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$a Kizek, Rene $u Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic; Department of Chemistry and Biochemistry, Mendel University in Brno, Brno, Czech Republic.
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$a Chmelik, Radim $u Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic; Institute of Physical Engineering, Faculty of Mechanical Engineering, Brno University of Technology, Brno, Czech Republic.
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$a Masarik, Michal $u Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic; Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic.
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