Nuclear Morphometric Analysis (NMA): Screening of Senescence, Apoptosis and Nuclear Irregularities

PLoS One

Eduardo C. Filippi-Chiela¹  Guido Lenz¹  Sidia Maria Callegari-Jacques²  Bruno Jurkovski³  Manuel M. Oliveira³  Vinicius Duval da Silva⁴ 

[1] Department of Biophysics, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil;Graduate Program in Genetics and Molecular Biology, Department of Statistics, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil;Institute of Informatics, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Rio Grande do Sul, Brazil;Sao Lucas Hospital – Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre, Rio Grande do Sul, Brazil

关键词: Apoptosis;    Cell staining;    Morphometry;    Mitosis;    DAPI staining;    HeLa cells;    Condensation;    Nuclear staining;   

DOI  :  10.1371/journal.pone.0042522

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Several cellular mechanisms affect nuclear morphology which can therefore be used to assess certain processes. Here, we present an analytic tool to quantify the number of cells in a population that present characteristics of senescence, apoptosis or nuclear irregularities through nuclear morphometric analysis. The tool presented here is based on nuclear image analysis and evaluation of size and regularity of adhered cells in culture. From 46 measurements of nuclear morphometry, principal component analysis filtered four measurements that best separated regular from irregular nuclei. These measurements, namely aspect, area box, radius ratio and roundness were combined into a single nuclear irregularity index (NII). Normal nuclei are used to set the parameters for a given cell type, and different nuclear phenotypes are separated in an area versus NII plot. The tool was validated with β-gal staining for senescence and annexin or caspases inhibitor for apoptosis as well as several treatments that induce different cellular phenotypes. This method provides a direct and objective way of screening normal, senescent, apoptotic and nuclear irregularities which may occur during failed mitosis or mitotic catastrophe, which may be very useful in basic and clinical research.

【 授权许可】

CC BY