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Liquid Handling Optimization in High-Throughput Biodosimetry Tool

机译:高通量生物剂量测定工具中的液体处理优化

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摘要

Due to the need of high-speed and efficient biodosimetric assays for triage and therapy in the event of radiological or nuclear attack, a robotically based automated biodosimetry tool (RABiT) has been developed over the past few years. Adapting the micronucleus assay from filter plates to V-shaped plates presented challenges in the liquid handling, namely, cell splashing out of the V-shaped well plate during the cell harvesting, poor cell distribution on the bottom of the image plate during the dispensing, and cell loss from the image plate during the aspiration in the liquid handling process. Experimental and numerical investigations were carried out to better understand the phenomena and mitigate the problems. Surface tension and contact angle among the fluids and the plate wall were accounted for in the discrete and multiphase numerical models. Experimental conditions were optimized based on the numerical results showing the relationship between nozzle speed and amount of splashed liquid, and the relationship between aspiration speed and number of escaped cells. Using these optimized parameters, numbers of micronuclei in binucleated cells showed the same dose dependence in the RABiT-prepared samples as those in the manually prepared ones. Micronucleus assay protocol was fully realized on RABiT.
机译:由于在放射或核袭击事件中需要进行快速高效的生物剂量测定以进行分类和治疗,因此在过去几年中已经开发了基于机器人的自动生物剂量测定工具(RABiT)。从过滤板到V形板的微核化验方法面临着液体处理方面的挑战,即在细胞收获过程中细胞从V形孔板中溅出,在分配过程中图像板底部的细胞分布不佳,在液体处理过程中,在抽吸过程中,图像板上的细胞损失和图像损失。为了更好地理解现象并减轻问题,进行了实验和数值研究。在离散和多相数值模型中考虑了流体与板壁之间的表面张力和接触角。根据数值结果优化了实验条件,数值结果显示了喷嘴速度与飞溅液体量之间的关系,以及抽吸速度与逃逸细胞数之间的关系。使用这些优化的参数,在RABiT制备的样品中,双核细胞中的微核数量显示出与手动制备的样品相同的剂量依赖性。在RABiT上完全实现了微核分析方案。

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