Measurement of apoptosis by SCAN©, a system for counting and analysis of fluorescently labelled nuclei

Neta Shlezinger, Elad Eizner, Stas Dubinchik, Anna Minz-Dub, Rachel Tetroashvili, Adi Reider, Amir Sharon*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Apoptosis-like programmed cell death (A-PCD) is a universal process common to all types of eukaryotic organisms. Because A-PCD-associated processes are conserved, it is possible to define A-PCD by a standard set of markers. Many of the popular methods to measure A-PCD make use of fluorescent ligands that change in intensity or cellular localization during A-PCD. In single cell organisms, it is possible to quantify levels of A-PCD by scoring the number of apoptotic cells using flow cytometry instruments. In a multicellular organism, quantification of A-PCD is more problematic due to the complex nature of the tissue. The situation is further complicated in filamentous fungi, in which nuclei are divided between compartments, each containing a number of nuclei, which can also migrate between the compartments. We developed SCAN©, a System for Counting and Analysis of Nuclei, and used it to measure A-PCD according to two markers – chromatin condensation and DNA strand breaks. The package includes three modules designed for counting the number of nuclei in multi-nucleated domains, scoring the relative number of nuclei with condensed chromatin, and calculating the relative number of nuclei with DNA strand breaks. The method provides equal or better results compared with manual counting, the analysis is fast and can be applied on large data sets. While we demonstrated the utility of the software for measurement of A-PCD in fungi, the method is readily adopted for measurement of A-PCD in other types of multicellular specimens.

Original languageAmerican English
Pages (from-to)406-415
Number of pages10
JournalMicrobial Cell
Volume1
Issue number12
DOIs
StatePublished - Dec 2014
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2014 Shlezinger et al.

Keywords

  • Apoptosis
  • DNA condensation
  • Fungi
  • PCD
  • TUNEL

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