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A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis
A novel dye-based method for measuring cell death in image-based screens is presented. Unlike conventional high- and medium-throughput cell death assays that measure only one form of cell death accurately, using multivariate analysis of micrographs of cells stained with the inexpensive mix, red dye...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Mary Ann Liebert, Inc.
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652219/ https://www.ncbi.nlm.nih.gov/pubmed/26422066 http://dx.doi.org/10.1089/adt.2015.661 |
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author | Collins, Tony J. Ylanko, Jarkko Geng, Fei Andrews, David W. |
author_facet | Collins, Tony J. Ylanko, Jarkko Geng, Fei Andrews, David W. |
author_sort | Collins, Tony J. |
collection | PubMed |
description | A novel dye-based method for measuring cell death in image-based screens is presented. Unlike conventional high- and medium-throughput cell death assays that measure only one form of cell death accurately, using multivariate analysis of micrographs of cells stained with the inexpensive mix, red dye nonyl acridine orange, and a nuclear stain, it was possible to quantify cell death induced by a variety of different agonists even without a positive control. Surprisingly, using a single known cytotoxic agent as a positive control for training a multivariate classifier allowed accurate quantification of cytotoxicity for mechanistically unrelated compounds enabling generation of dose–response curves. Comparison with low throughput biochemical methods suggested that cell death was accurately distinguished from cell stress induced by low concentrations of the bioactive compounds Tunicamycin and Brefeldin A. High-throughput image-based format analyses of more than 300 kinase inhibitors correctly identified 11 as cytotoxic with only 1 false positive. The simplicity and robustness of this dye-based assay makes it particularly suited to live cell screening for toxic compounds. |
format | Online Article Text |
id | pubmed-4652219 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Mary Ann Liebert, Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-46522192015-12-02 A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis Collins, Tony J. Ylanko, Jarkko Geng, Fei Andrews, David W. Assay Drug Dev Technol Original Articles A novel dye-based method for measuring cell death in image-based screens is presented. Unlike conventional high- and medium-throughput cell death assays that measure only one form of cell death accurately, using multivariate analysis of micrographs of cells stained with the inexpensive mix, red dye nonyl acridine orange, and a nuclear stain, it was possible to quantify cell death induced by a variety of different agonists even without a positive control. Surprisingly, using a single known cytotoxic agent as a positive control for training a multivariate classifier allowed accurate quantification of cytotoxicity for mechanistically unrelated compounds enabling generation of dose–response curves. Comparison with low throughput biochemical methods suggested that cell death was accurately distinguished from cell stress induced by low concentrations of the bioactive compounds Tunicamycin and Brefeldin A. High-throughput image-based format analyses of more than 300 kinase inhibitors correctly identified 11 as cytotoxic with only 1 false positive. The simplicity and robustness of this dye-based assay makes it particularly suited to live cell screening for toxic compounds. Mary Ann Liebert, Inc. 2015-11-01 /pmc/articles/PMC4652219/ /pubmed/26422066 http://dx.doi.org/10.1089/adt.2015.661 Text en © Collins et al. 2015; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Articles Collins, Tony J. Ylanko, Jarkko Geng, Fei Andrews, David W. A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title | A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title_full | A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title_fullStr | A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title_full_unstemmed | A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title_short | A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis |
title_sort | versatile cell death screening assay using dye-stained cells and multivariate image analysis |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652219/ https://www.ncbi.nlm.nih.gov/pubmed/26422066 http://dx.doi.org/10.1089/adt.2015.661 |
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