RABiT-II: A Fully-Automated Micronucleus Assay System with Shortened Time to Result
RABiT-II: A Fully-Automated Micronucleus Assay System with Shortened Time to Result
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DOI：
10.1667/rr15215.1
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发表时间：
2019-03-01
影响因子：
3.4
通讯作者：
Brenner, David J.
中科院分区：
文献类型：
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作者：
Repin, Mikhail;Pampou, Sergey;Brenner, David J.
在这项工作中，我们描述了一个完全自动化的微核（CBMN）测定，结果显着缩短的时间，需要快速高通量的生物剂量估计的辐射剂量从小体积的人血液样本的动机。快速自动生物剂量测定工具（RABiT-II）目前由两个商业自动化系统组成：PerkinElmer细胞：：explorer工作站和GE Healthcare IN细胞分析仪2000成像仪。用0（对照）、0.5、1.5、2.5、3.5或4.5戈伊离体γ射线照射来自8名健康志愿者的血样（30 μ l），并在RABiT-II系统上的96孔板中全自动处理。总细胞培养时间为54小时，总测定时间为3天。将DAPI染色的固定样品在IN Cell Analyzer 2000上成像，使用GE Healthcare IN Cell Developer 1.9版进行全自动图像分析。建立CBMN剂量-反应校准曲线，之后评估系统预测已知剂量的能力。在97%的样本中，来自两个供体的辐照样本的各种辐射剂量估计在真实剂量的20%以内（+/- 0.5戈伊，低于2戈伊），某些5戈伊辐照样本的剂量被低估高达25%。总之，这项工作的结果表明，加速CBMN测定可以在96孔板中使用商业生物技术机器人系统以高通量形式自动化，为辐射暴露提供快速可靠的生物测定。(C)2019辐射研究学会
In this work, we describe a fully automated cytokinesis-block micronucleus (CBMN) assay with a significantly shortened time to result, motivated by the need for rapid high-throughput biodosimetric estimation of radiation doses from small-volume human blood samples. The Rapid Automated Biodosimetry Tool (RABiT-II) currently consists of two commercial automated systems: a PerkinElmer cell:: explorer Workstation and a GE Healthcare IN Cell Analyzer 2000 Imager. Blood samples (30 mu l) from eight healthy volunteers were gamma-ray irradiated ex vivo with 0 (control), 0.5, 1.5, 2.5, 3.5 or 4.5 Gy and processed with full automation in 96-well plates on the RABiT-II system. The total cell culture time was 54 h and total assay time was 3 days. DAPI-stained fixed samples were imaged on an IN Cell Analyzer 2000 with fully-automated image analysis using the GE Healthcare IN Cell Developer Toolbox version 1.9. A CBMN dose-response calibration curve was established, after which the capability of the system to predict known doses was assessed. Various radiation doses for irradiated samples from two donors were estimated within 20% of the true dose (+/- 0.5 Gy below 2 Gy) in 97% of the samples, with the doses in some 5 Gy irradiated samples being underestimated by up to 25%. In summary, the findings from this work demonstrate that the accelerated CBMN assay can be automated in a high-throughput format, using commercial biotech robotic systems, in 96-well plates, providing a rapid and reliable bioassay for radiation exposure. (C) 2019 by Radiation Research Society