Analysis of chromosome damage for biodosimetry using imaging flow cytometry

Beaton, Lindsay A.; Ferrarotto, C.; Kutzner, B. C.; McNamee, James P.; Bellier, Pascale V.; Wilkins, Ruth

doi:10.1016/j.mrgentox.2013.04.002

Cited by 16 publications

(10 citation statements)

References 10 publications

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“…The EDF option on the ISX allows light from different focal planes to be imaged on the detector plane simultaneously. The EDF is a useful option that allows structures and probes, such as individual chromosomes in suspension (54,55) or cH2AX foci (56,57) to be focused onto a single two-dimensional image. This improves the ability to count spots at different focus depths within the cell.…”

Section: Discussionmentioning

confidence: 99%

Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer

Rodrigues¹

2018

Cytometry Pt A

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The in vitro micronucleus (MN) assay is a well‐established test for evaluating genotoxicity and cytotoxicity. The use of manual microscopy to perform the assay can be laborious and often suffers from user subjectivity and interscorer variability. Automated methods including slide‐scanning microscopy and conventional flow cytometry have been developed to eliminate scorer bias and improve throughput. However, these methods possess several limitations such as lack of cytoplasmic visualization using slide‐scanning microscopy and the inability to visually confirm the legitimacy of MN or storage of image data for re‐evaluation using flow cytometry. The ImageStreamX® MK II (ISX) imaging flow cytometer has been demonstrated to overcome all of these limitations. The ISX combines the speed, statistical robustness, and rare event capture capability of conventional flow cytometry with high resolution fluorescent imagery of microscopy and possesses the ability to store all collected image data. This paper details the methodology developed to perform the in vitro MN assay in human lymphoblastoid TK6 cells on the ISX. High resolution images of micronucleated mono‐ and bi‐nucleated cells as well as polynucleated cells can be acquired at a high rate of capture. All images can then be automatically identified, categorized and enumerated in the data analysis software that accompanies the ImageStream, allowing for the scoring of both genotoxicity and cytotoxicity. The results demonstrate that statistically significant increases in MN frequency when compared with solvent controls can be detected at varying levels of cytotoxicity following exposure to well‐known aneugens and clastogens. This work demonstrates a fully automated method for performing the in vitro micronucleus assay on the ISX imaging flow cytometry platform. © 2018 The Author. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.

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Section: Discussionmentioning

confidence: 99%

Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer

Rodrigues¹

2018

Cytometry Pt A

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“…To identify spots in a cell, the use of Spot mask might be the best option [13, 14, 15]. This mask extracts bright or dark regions of an object, and can be further refined with Peak and/or Range masks.…”

Section: Troubleshooting Masksmentioning

confidence: 99%

Masks in imaging flow cytometry

Dominical

Samsel

McCoy

2017

Methods

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Data analysis in imaging flow cytometry incorporates elements of flow cytometry together with other aspects of morphological analysis of images. A crucial early step in this analysis is the creation of a mask to distinguish the portion of the image upon which further examination of specified features can be performed. Default masks are provided by the manufacturer of the imaging flow cytometer but additional custom masks can be created by the individual user for specific applications. Flawed or inaccurate masks can have a substantial negative impact on the overall analysis of a sample, thus great care must be taken to ensure the accuracy of masks. Here we discuss various types of masks and cite examples of their use. Furthermore we provide our insight for how to approach selecting and assessing the optimal mask for a specific analysis.

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“…Frequency of occurrence of cells with dumbbell-shaped nuclei depends on the dose of irradiation and this correlation is described by a linearquadratic function with the following equation: y = 0.003+0.014x +0.005x 2 [31]. We would like to emphasize that the simplest biomarkers (nuclei abnormalities) we suggest for radiation exposure detection in vivo is not an alternative to classical radiation biomarkers in vitro [17], as well as to other modern approaches [2][3][4][5][44][45][46]. Detection of chromosomal aberrations (dicentrics) and the cytochalasine B micronucleus and "Cytome" methods are reliable bioindication and biodosimetry methods.…”

Section: Dumbbell-shaped Nucleimentioning

confidence: 99%

Nuclear Abnormalities of Lymphocytes as the Simplest Markers for Bioindication Test in Case of Mass Casualty Events Involving Radiation Exposure

Kravtsov¹,

Livanova²,

Starkova³

2017

Emerg Med

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Radiation exposure leads to a large number of victims who seek medical help in the first hours. To provide medical care, it is necessary to correctly establish the fact and dose of radiation by bioindication and biodosimetry methods. Lymphocytes containing nuclear anomalies are easily detected cells of peripheral blood and are suitable as objects of bioindication in the case of radioactive exposure to a large number of people. Among these anomalies we distinguish micronuclei, "tailed" nuclei, nucleoplasmatic bridges, dumbbell-shaped nuclei, etc. This review observes the main types of these nuclear anomalies of lymphocytes in the light of their common origin from dicentric chromosomes. We recommend using these alterations in peripheral blood lymphocytes nuclei as the simplest biomarkers in the framework of bioindication tests when it is necessary to work with a large number of victims.

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Analysis of chromosome damage for biodosimetry using imaging flow cytometry

Cited by 16 publications

References 10 publications

Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer

Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer

Masks in imaging flow cytometry

Nuclear Abnormalities of Lymphocytes as the Simplest Markers for Bioindication Test in Case of Mass Casualty Events Involving Radiation Exposure

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Analysis of chromosome damage for biodosimetry using imaging flow cytometry

Cited by 16 publications

References 10 publications

Automation of the in vitro micronucleus assay using the Imagestream® imaging flow cytometer

Automation of the in vitro micronucleus assay using the Imagestream® imaging flow cytometer

Masks in imaging flow cytometry

Nuclear Abnormalities of Lymphocytes as the Simplest Markers for Bioindication Test in Case of Mass Casualty Events Involving Radiation Exposure

Contact Info

Product

Resources

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Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer

Automation of the in vitro micronucleus assay using the Imagestream^® imaging flow cytometer