Multimodal holographic microscopy: distinction between apoptosis and oncosis

PLoS One. 2015 Mar 24;10(3):e0121674. doi: 10.1371/journal.pone.0121674. eCollection 2015.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Cell Line, Tumor
  • Cell Survival
  • Holography / methods*
  • Humans
  • Microscopy, Fluorescence / methods*
  • Multimodal Imaging / methods*
  • Necrosis
  • Phenotype
  • Time Factors

Grants and funding

The financial support from CEITEC CZ.1.05/1.1.00/02.0068 and project for conceptual development of research organization (Faculty of Medicine, Masaryk university - ROZV/24/LF5/2014) is greatly acknowledged. Part of the work was carried out with the support of core facilities of CEITEC – Central European Institute of Technology under CEITEC-open access project, ID number LM2011020, funded by the Ministry of Education, Youth and Sports of the Czech Republic under the activity “Projects of major infrastructures for research, development and innovations”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.