Cytokinesis-block micronucleus cytome assay

Fenech, Michael

doi:10.1038/nprot.2007.77

Cited by 1,741 publications

(1,583 citation statements)

References 104 publications

Supporting

Mentioning

1,444

Contrasting

Unclassified

111

Order By: Relevance

“…Finally, the Range mask was created using the MN mask to allow proper identification of MN between 8 and 120 pixels (0.89 and 13.3 μm 2 ). This range is slightly different than that proposed by published scoring criteria which indicates that MN should have areas that vary between 1/256th and 1/9th of the area of the main nuclei 9, 46. Normally this would correspond to a MN range between 0.40 and 11.1 μm 2 since the mean area of the main nuclei was approximately 100 μm 2 .…”

Section: Resultsmentioning

confidence: 65%

“…Using the merged files, truth populations consisting of 50 positive and 50 negative hand‐tagged cells were created for all cell types of interest (MONO, BN, and POLY cells). As demonstrated in Figure 1, the positive cell populations (middle column) have visible cytoplasmic material, highly circular and in‐focus nuclei and, in the case of multinucleated cells, nuclei that are similar in size and staining intensity relative to one another as per the scoring criteria described by Fenech et al 9, 46. On the other hand, the negative population (right hand column) contains cells with abnormal nuclear morphology, overlapping nuclei or nuclei of differing intensity, area, circularity and aspect ratios.…”

Section: Resultsmentioning

confidence: 89%

“…Therefore, it is often necessary to perform the assay both in the presence and absence of Cyt‐B, which increases the number of samples that must be evaluated. In recent years, several protocols and exposure schedules to perform the MN assay have been developed and validated in a number of recommended cell types 9, 10, 11, 12. Current forms of the assay are able to detect the presence of clastogens and aneugens in both rodent and human derived cell lines as well as in human peripheral blood lymphocytes 13, 14, 15, 16, 17.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Rodrigues¹

2018

Cytometry Pt A

View full text Add to dashboard Cite

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.

show abstract

Section: Resultsmentioning

confidence: 65%

Section: Resultsmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

Rodrigues¹

2018

Cytometry Pt A

View full text Add to dashboard Cite

show abstract

“…All slides were coded before scoring. The binucleated cells (BNC) were selected according to criteria described by Umegaki and Fenech [2000] and Fenech [2007]. Apoptotic and necrotic cells were included in the total cell number; however micronuclei were not scored in these cells.…”

Section: Microscopic Evaluation and Micronuclei (Mn) Scoringmentioning

confidence: 99%

Genotoxic effects of three selected black toner powders and their dimethyl sulfoxide extracts in cultured human epithelial A549 lung cells in vitro

Gminski

Decker

Heinz

et al. 2010

Environ and Mol Mutagen

View full text Add to dashboard Cite

Until now, the adverse effects of toner powders on humans have been considered to be minimal. However, several recent reports have suggested possible significant adverse health effects from toner dust inhalation. The aim of this study was to evaluate the genotoxic potential of black toner powders in vitro. For the study of DNA damage, A549 cells were exposed to toner-powder suspensions and to their DMSO extracts, and then subjected to the comet assay and to the in-vitro cytokinesis block micronucleus test (CB-MNvit). Cytotoxic effects of the toner samples were assessed by the erythrosin B assay. Furthermore, size, shape, and composition of the toner powders were investigated. None of the three toner powders or their DMSO extracts reduced cell viability; however, they did induce DNA damage and formed micronuclei at concentrations from 80 to 400 lg cm 22 , although to a varying extent. All toner powders contain considerable amounts of the pigments carbon black and magnetite (Fe 3 O 4 ) as well as small amounts of polycyclic aromatic hydrocarbons (PAHs). The overall results of our invitro study suggest that the investigated toner-powder samples are not cytotoxic but genotoxic. From the results of the physical and chemical characterization, we conclude that metals and metalloids as components of magnetite, or PAHs as components of the carbon-bearing material, are responsible for the genotoxic effects. Further research is necessary to determine the relevance of these in-vitro observations for private and occupational toner powder exposure.

show abstract

“…The analysis of MN was performed using a light microscope (Nikon E50i, Otawara, Tochigi, Japan) at a magnification of 9400 following the criteria for MN scoring in binucleated (BN) cells as described by Fenech (2007). Three thousand BN cells per concentration (one thousand per donor) were examined for MN scoring and 500 cells per concentration were examined to determine the number of cells with 1, 2, 3 and 4 nuclei.…”

Section: Mn Analysismentioning

confidence: 99%

Evaluation of in vitro antioxidant, antimicrobial, genotoxic and anticancer activities of lichen Cetraria islandica

et al. 2014

View full text Add to dashboard Cite

In this study, the antioxidant, antimicrobial, genotoxic and anticancer activities of Cetraria islandica methanol extract were determined by using free radical and superoxide anion scavenging activity, reducing power, determination of total phenolic compounds and flavonoid contents, broth microdilution minimal inhibitory concentration against five bacterial and five fungal species, cytokinesis block micronucleus (MN) assay on peripheral blood lymphocytes (PBLs) and the microculture tetrazolium test on FemX (human melanoma) and LS174 (human colon carcinoma) cell lines. As a result of the study, we found that C. islandica methanol extract exhibited moderate free-radical-scavenging activity with IC 50 values 678.38 lg/ml. Moreover, the tested extract had effective reducing power and superoxide anion radical scavenging. The minimal inhibitory concentration values against the tested microorganisms ranged from 0.312 to 5 mg/ml. The extract increased MN frequency in a dose dependent manner, but it was significant in higher tested concentrations (50, 100 and 200 lg/ml). No significant differences were observed between NDI values in all treatments and untreated PBLs. In addition, the tested extract had strong anticancer activity towards both cell lines with IC 50 values of 22.68 and 33.74 lg/ml. It can be concluded that the tested extract exhibited a certain level of in vitro antioxidant, antimicrobial, genotoxic and anticancer activities.

show abstract

Cytokinesis-block micronucleus cytome assay

Cited by 1,741 publications

References 104 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

Genotoxic effects of three selected black toner powders and their dimethyl sulfoxide extracts in cultured human epithelial A549 lung cells in vitro

Evaluation of in vitro antioxidant, antimicrobial, genotoxic and anticancer activities of lichen Cetraria islandica

Contact Info

Product

Resources

About

Cytokinesis-block micronucleus cytome assay

Cited by 1,741 publications

References 104 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

Genotoxic effects of three selected black toner powders and their dimethyl sulfoxide extracts in cultured human epithelial A549 lung cells in vitro

Evaluation of in vitro antioxidant, antimicrobial, genotoxic and anticancer activities of lichen Cetraria islandica

Contact Info

Product

Resources

About

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