Automated sample processing for flow cytometry

Jasenka Memisevic, L. Shriver-Lake, P. Howell, J. Golden, N. Hashemi, Kirsten B. Jackson, F. Ligler
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Abstract

The utility of pathogen and toxin detection systems depends not only upon sensitivity, specificity and capability for multiplexed recognition, but also on access at the point of need, ease of use, and response time. Combining microfluidics and optical biosensors facilitates miniaturization and automation, while careful design makes it possible to test variable quantities of complex matrices such as food (mL) and clinical samples (µL). We report on progress towards an optofluidic system that combines a rotating magnet trap for automated sample processing and a microflow cytometer capable of 4-color analysis to achieve multi-analyte diagnostics. Sample and antibody-coated magnetic beads are introduced into a moving magnetic field created by magnets rotating in the opposite direction of the flow. The color-coded magnetic beads with captured target remain in suspension while fluorescent tracer reagents are introduced to the flow, thus optimizing the binding kinetics and minimizing aggregation. Magnetic beads with bound target and tracer reagents are released for multiplexed analysis by reversing the direction of spinning magnets.
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用于流式细胞术的自动样品处理
病原体和毒素检测系统的效用不仅取决于敏感性、特异性和多重识别的能力,还取决于在需要时的可及性、易用性和反应时间。结合微流体和光学生物传感器促进了小型化和自动化,而精心设计使得可以测试可变数量的复杂基质,如食品(mL)和临床样品(µL)。我们报告了一种光流体系统的进展,该系统结合了用于自动样品处理的旋转磁铁陷阱和能够进行4色分析的微流式细胞仪,以实现多分析物诊断。样品和抗体包覆的磁珠被引入到一个移动的磁场中,磁场是由与流动方向相反的磁铁旋转产生的。带有捕获目标的彩色编码磁珠保持悬浮状态,而荧光示踪试剂被引入到流动中,从而优化结合动力学并最大限度地减少聚集。通过反转旋转磁体的方向,释放带有结合靶和示踪试剂的磁珠进行多路分析。
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