Center for Radiological Research.
Columbia Genome Center High-Throughput Screening Facility, Columbia University Medical Center, New York, New York 10032.
Radiat Res. 2019 Sep;192(3):311-323. doi: 10.1667/RR15266.1. Epub 2019 Jul 11.
We developed a fully-automated dicentric chromosome assay (DCA) in multiwell plates. All operations, from sample loading to chromosome scoring, are performed, without human intervention, by the second-generation Rapid Automated Biodosimetry Tool II (RABiT-II) robotic system, a plate imager and custom software, FluorQuantDic. The system requires small volumes of blood (30 µl per individual) to determine radiation dose received as a result of a radiation accident or terrorist attack. To visualize dicentrics in multiwell plates, we implemented a non-classical protocol for centromere FISH staining at 37°C. The RABiT-II performs rapid analysis of chromosomes after extracting them from metaphase cells. With the use of multiwell plates, many samples can be screened at the same time. Thus, the RABiT-II DCA provides an advantage during triage when risk-based stratification and medical management are required for a large population exposed to unknown levels of ionizing radiation.
我们开发了一种全自动的双着丝粒染色体分析（DCA）方法，可在多孔板中进行。所有操作，从样本加载到染色体评分，都由第二代 Rapid Automated Biodosimetry Tool II（RABiT-II）机器人系统、平板成像仪和自定义软件 FluorQuantDic 自动完成，无需人工干预。该系统只需少量血液（每人 30 µl）即可确定因辐射事故或恐怖袭击而接受的辐射剂量。为了在多孔板中可视化双着丝粒，我们实施了一种非经典的 37°C 着丝粒 FISH 染色方案。RABiT-II 在从中期细胞中提取染色体后，快速进行染色体分析。通过使用多孔板，可以同时筛选多个样本。因此，当需要对暴露于未知水平电离辐射的大量人群进行基于风险的分层和医疗管理时，RABiT-II DCA 可用于分诊。
