The in vitro micronucleus assay using imaging flow cytometry and deep learning

Abstract The in vitro micronucleus (MN) assay is a well-established assay for quantification of DNA damage, and is required by regulatory bodies worldwide to screen chemicals for genetic toxicity. The MN assay is performed in two variations: scoring MN in cytokinesis-blocked binucleated cells or dir...

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Autores principales:	Matthew A. Rodrigues, Christine E. Probst, Artiom Zayats, Bryan Davidson, Michael Riedel, Yang Li, Vidya Venkatachalam
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/de6553bcb0b54e6eabcf48cd6ad699a3
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spelling	oai:doaj.org-article:de6553bcb0b54e6eabcf48cd6ad699a32021-12-02T15:53:02ZThe in vitro micronucleus assay using imaging flow cytometry and deep learning10.1038/s41540-021-00179-52056-7189https://doaj.org/article/de6553bcb0b54e6eabcf48cd6ad699a32021-05-01T00:00:00Zhttps://doi.org/10.1038/s41540-021-00179-5https://doaj.org/toc/2056-7189Abstract The in vitro micronucleus (MN) assay is a well-established assay for quantification of DNA damage, and is required by regulatory bodies worldwide to screen chemicals for genetic toxicity. The MN assay is performed in two variations: scoring MN in cytokinesis-blocked binucleated cells or directly in unblocked mononucleated cells. Several methods have been developed to score the MN assay, including manual and automated microscopy, and conventional flow cytometry, each with advantages and limitations. Previously, we applied imaging flow cytometry (IFC) using the ImageStream® to develop a rapid and automated MN assay based on high throughput image capture and feature-based image analysis in the IDEAS® software. However, the analysis strategy required rigorous optimization across chemicals and cell lines. To overcome the complexity and rigidity of feature-based image analysis, in this study we used the Amnis® AI software to develop a deep-learning method based on convolutional neural networks to score IFC data in both the cytokinesis-blocked and unblocked versions of the MN assay. We show that the use of the Amnis AI software to score imagery acquired using the ImageStream® compares well to manual microscopy and outperforms IDEAS® feature-based analysis, facilitating full automation of the MN assay.Matthew A. RodriguesChristine E. ProbstArtiom ZayatsBryan DavidsonMichael RiedelYang LiVidya VenkatachalamNature PortfolioarticleBiology (General)QH301-705.5ENnpj Systems Biology and Applications, Vol 7, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Matthew A. Rodrigues Christine E. Probst Artiom Zayats Bryan Davidson Michael Riedel Yang Li Vidya Venkatachalam The in vitro micronucleus assay using imaging flow cytometry and deep learning
description	Abstract The in vitro micronucleus (MN) assay is a well-established assay for quantification of DNA damage, and is required by regulatory bodies worldwide to screen chemicals for genetic toxicity. The MN assay is performed in two variations: scoring MN in cytokinesis-blocked binucleated cells or directly in unblocked mononucleated cells. Several methods have been developed to score the MN assay, including manual and automated microscopy, and conventional flow cytometry, each with advantages and limitations. Previously, we applied imaging flow cytometry (IFC) using the ImageStream® to develop a rapid and automated MN assay based on high throughput image capture and feature-based image analysis in the IDEAS® software. However, the analysis strategy required rigorous optimization across chemicals and cell lines. To overcome the complexity and rigidity of feature-based image analysis, in this study we used the Amnis® AI software to develop a deep-learning method based on convolutional neural networks to score IFC data in both the cytokinesis-blocked and unblocked versions of the MN assay. We show that the use of the Amnis AI software to score imagery acquired using the ImageStream® compares well to manual microscopy and outperforms IDEAS® feature-based analysis, facilitating full automation of the MN assay.
format	article
author	Matthew A. Rodrigues Christine E. Probst Artiom Zayats Bryan Davidson Michael Riedel Yang Li Vidya Venkatachalam
author_facet	Matthew A. Rodrigues Christine E. Probst Artiom Zayats Bryan Davidson Michael Riedel Yang Li Vidya Venkatachalam
author_sort	Matthew A. Rodrigues
title	The in vitro micronucleus assay using imaging flow cytometry and deep learning
title_short	The in vitro micronucleus assay using imaging flow cytometry and deep learning
title_full	The in vitro micronucleus assay using imaging flow cytometry and deep learning
title_fullStr	The in vitro micronucleus assay using imaging flow cytometry and deep learning
title_full_unstemmed	The in vitro micronucleus assay using imaging flow cytometry and deep learning
title_sort	in vitro micronucleus assay using imaging flow cytometry and deep learning
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/de6553bcb0b54e6eabcf48cd6ad699a3
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The in vitro micronucleus assay using imaging flow cytometry and deep learning

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