A flow cytometry based in vitro micronucleus assay in TK6 cells-Validation using early stage pharmaceutical development compounds

2010 ◽  
Vol 52 (5) ◽  
pp. 363-372 ◽  
Author(s):  
Magdalena Lukamowicz ◽  
Katherine Woodward ◽  
Micheline Kirsch-Volders ◽  
Willi Suter ◽  
Azeddine Elhajouji
Keyword(s):  
Flow Cytometry ◽  
Early Stage ◽  
Micronucleus Assay ◽  
Pharmaceutical Development ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

Performance of the flow cytometry-based in vitro micronucleus assay in a validation study

2016 ◽  
Vol 258 ◽  
pp. S316
Author(s):  
D.J. Smart ◽  
F. Helbling ◽  
P. Blandine ◽  
M. Damian ◽  
V. Patrick
Keyword(s):  
Flow Cytometry ◽  
Validation Study ◽  
Micronucleus Assay ◽  
In Vitro Micronucleus Assay

scholarly journals Inter-laboratory automation of the in vitro micronucleus assay using imaging flow cytometry and deep learning

2021 ◽  
Author(s):  
John W. Wills ◽  
Jatin R. Verma ◽  
Benjamin J. Rees ◽  
Danielle S. G. Harte ◽  
Qiellor Haxhiraj ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Deep Learning ◽  
Image Classification ◽  
Micronucleus Assay ◽  
Scoring Method ◽  
Safety Testing ◽  
Imaging Flow Cytometry ◽  
Unseen Data ◽  
In Vitro Micronucleus Assay

AbstractThe in vitro micronucleus assay is a globally significant method for DNA damage quantification used for regulatory compound safety testing in addition to inter-individual monitoring of environmental, lifestyle and occupational factors. However, it relies on time-consuming and user-subjective manual scoring. Here we show that imaging flow cytometry and deep learning image classification represents a capable platform for automated, inter-laboratory operation. Images were captured for the cytokinesis-block micronucleus (CBMN) assay across three laboratories using methyl methanesulphonate (1.25–5.0 μg/mL) and/or carbendazim (0.8–1.6 μg/mL) exposures to TK6 cells. Human-scored image sets were assembled and used to train and test the classification abilities of the “DeepFlow” neural network in both intra- and inter-laboratory contexts. Harnessing image diversity across laboratories yielded a network able to score unseen data from an entirely new laboratory without any user configuration. Image classification accuracies of 98%, 95%, 82% and 85% were achieved for ‘mononucleates’, ‘binucleates’, ‘mononucleates with MN’ and ‘binucleates with MN’, respectively. Successful classifications of ‘trinucleates’ (90%) and ‘tetranucleates’ (88%) in addition to ‘other or unscorable’ phenotypes (96%) were also achieved. Attempts to classify extremely rare, tri- and tetranucleated cells with micronuclei into their own categories were less successful (≤ 57%). Benchmark dose analyses of human or automatically scored micronucleus frequency data yielded quantitation of the same equipotent concentration regardless of scoring method. We conclude that this automated approach offers significant potential to broaden the practical utility of the CBMN method across industry, research and clinical domains. We share our strategy using openly-accessible frameworks.

Flow cytometric 96-well microplate-based in vitro micronucleus assay with human TK6 cells: Protocol optimization and transferability assessment

2013 ◽  
Vol 54 (3) ◽  
pp. 180-194 ◽  
Author(s):  
Steven M. Bryce ◽  
Svetlana L. Avlasevich ◽  
Jeffrey C. Bemis ◽  
Matthew Tate ◽  
Richard M. Walmsley ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Flow Cytometric ◽  
Protocol Optimization ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

scholarly journals Analysis of negative historical control group data from the in vitro micronucleus assay using TK6 cells

2018 ◽  
Vol 825 ◽  
pp. 40-50 ◽  
Author(s):  
David P. Lovell ◽  
Mick Fellows ◽  
Francesco Marchetti ◽  
Joan Christiansen ◽  
Azeddine Elhajouji ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Historical Control Group ◽  
Historical Control ◽  
Control Group ◽  
Group Data ◽  
Tk6 Cells ◽  
In Vitro Micronucleus Assay

Development and validation of an in vitro micronucleus assay platform in TK6 cells

2012 ◽  
Vol 746 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Zhanna Sobol ◽  
Michael L. Homiski ◽  
Donna A. Dickinson ◽  
Richard A. Spellman ◽  
Dingzhou Li ◽  
...  
Keyword(s):  
Micronucleus Assay ◽  
Tk6 Cells ◽  
Development And Validation ◽  
In Vitro Micronucleus Assay
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