دورية أكاديمية

Application of the Cytokinesis-Block Micronucleus Assay for High-Dose Exposures Using Imaging Flow Cytometry.

التفاصيل البيبلوغرافية
العنوان: Application of the Cytokinesis-Block Micronucleus Assay for High-Dose Exposures Using Imaging Flow Cytometry.
المؤلفون: 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.
المصدر: Cytogenetic & Genome Research; 2024, Vol. 163 Issue 3/4, p131-142, 12p
مصطلحات موضوعية: FLOW cytometry, NUCLEOLUS, CELL analysis, CELL imaging, GENETIC toxicology, CURVE fitting
مستخلص: 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. [ABSTRACT FROM AUTHOR]
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قاعدة البيانات: Complementary Index
الوصف
تدمد:14248581
DOI:10.1159/000532124