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https://hdl.handle.net/2440/102196
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Type: | Journal article |
Title: | Automation of the cytokinesis-block micronucleus cytome assay by laser scanning cytometry and its potential application in radiation biodosimetry |
Author: | Francois, M. Hochstenbach, K. Leifert, W. Fenech, M. |
Citation: | BioTechniques: the journal of laboratory technology for bioresearch, 2014; 57(6):309-312 |
Publisher: | BioTechniques |
Issue Date: | 2014 |
ISSN: | 0736-6205 1940-9818 |
Statement of Responsibility: | M. François, K. Hochstenbach, W. Leifert and M.F. Fenech |
Abstract: | Here we describe the adaptation of laser scanning cytometry (LSC) to measure micronuclei (MN) automatically in lymphocytes. MN frequencies were determined in irradiated human lymphocytes using the cytokinesis-block technique, and the results from LSC were compared with visual scoring results obtained from slides of cells stained using Fast Green and 4´,6-diamidino-2-phenylindole (DAPI). This fluorescent approach allowed clear identification of binucleated cells and detection of MN. The dose responses measured visually and by LSC showed similar trends and correlated positively (r = 0.9689; P < 0.0001). Highcontent analysis was developed to further automatically score MN within mono-, tri- and tetra-nucleated cells and to determine the nuclear division index and nuclear circularity values. The high-throughput nature of LSC can provide unique advantages in future DNA damage diagnostics in experimental and epidemiological studies. Importantly, it allows for co-detection of other biomarkers of interest within a single lymphocyte, and further development of this capability is anticipated. |
Keywords: | Micronucleus assay; automation; laser scanning cytometry |
Rights: | Copyright States unknown |
DOI: | 10.2144/000114239 |
Published version: | http://dx.doi.org/10.2144/000114239 |
Appears in Collections: | Aurora harvest 7 Medical Sciences publications |
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