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Application of the Cytokinesis-Block Micronucleus Assay for High-Dose Exposures Using Imaging Flow Cytometry.
Beaton-Green, Lindsay A; Mayenburg, Jessica M; Marro, Leonora; Hassan, Eman M; Cuadros Sanchez, Sarita; Darwish, Riham; Lachapelle, Sylvie; Adam, Nadine; Burtt, Julie J; Van Den Hanenberg, Cyndi; Rodrigues, Matthew A; Wang, Qi; Brenner, David J; Turner, Helen C; Wilkins, Ruth C.
Afiliação
  • Beaton-Green LA; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Mayenburg JM; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Marro L; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Hassan EM; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Cuadros Sanchez S; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Darwish R; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Lachapelle S; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Adam N; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Burtt JJ; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Van Den Hanenberg C; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
  • Rodrigues MA; Cytek Biosciences, Seattle, Washington, USA.
  • Wang Q; Center for Radiological Research, Columbia University Medical Center, New York, New York, USA.
  • Brenner DJ; Center for Radiological Research, Columbia University Medical Center, New York, New York, USA.
  • Turner HC; Center for Radiological Research, Columbia University Medical Center, New York, New York, USA.
  • Wilkins RC; Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario, Canada.
Cytogenet Genome Res ; 163(3-4): 131-142, 2023.
Article em En | MEDLINE | ID: mdl-37527635
The cytokinesis-block micronucleus assay is a well-established method to assess radiation-induced genetic damage in human cells. This assay has been adapted to imaging flow cytometry (IFC), allowing automated analysis of many cells, and eliminating the need to create microscope slides. Furthermore, to improve the efficiency of assay performance, a small-volume method previously developed was employed. Irradiated human blood samples were cultured, stained, and analyzed by IFC to produce images of the cells. Samples were run using both manual and 96-well plate automated acquisition. Multiple parameter-based image features were collected for each sample, and the results were compared to confirm that these acquisition methods are functionally identical. This paper details the multi-parametric analysis developed and the resulting calibration curves up to 10 Gy. The calibration curves were created using a quadratic random coefficient model with Poisson errors, as well as a logistic discriminant function. The curves were then validated with blinded, irradiated samples, using relative bias and relative mean square error. Overall, the accuracy of the dose estimates was adequate for triage dosimetry (within 1 Gy of the true dose) over 90% of the time for lower doses and about half the time for higher doses, with the lowest success rate between 5 and 6 Gy where the calibration curve reached its peak and there was the smallest change in MN/BNC with dose. This work describes the application of a novel multi-parametric analysis that fits the calibration curves and allows dose estimates up to 10 Gy, which were previously limited to 4 Gy. Furthermore, it demonstrates that the results from samples acquired manually and with the autosampler are functionally similar.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Radiometria / Citocinese Limite: Humans Idioma: En Revista: Cytogenet Genome Res Assunto da revista: GENETICA Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Canadá

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Radiometria / Citocinese Limite: Humans Idioma: En Revista: Cytogenet Genome Res Assunto da revista: GENETICA Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Canadá