Padmanabhan S, Sposito A, Yeh M, Everitt M, White I, DeVoe D L
Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, Maryland 20742, USA.
Department of Mechanical Engineering, University of Maryland, College Park, Maryland 20742, USA.
Biomicrofluidics. 2021 Jan 15;15(1):014103. doi: 10.1063/5.0039146. eCollection 2021 Jan.
The seamless integration of reagents into microfluidic devices can serve to significantly reduce assay complexity and cost for disposable diagnostics. In this work, the integration of multiplexed reagents into thermoplastic 2D microwell arrays is demonstrated using a scalable pin spotting technique. Using a simple and low-cost narrow-bore capillary spotting pin, high resolution deposition of concentrated reagents within the arrays of enclosed nanoliter-scale wells is achieved. The pin spotting method is further employed to encapsulate the deposited reagents with a chemically modified wax layer that serves to prevent disruption of the dried assay components during sample introduction through a shared microchannel, while also enabling temperature-controlled release after sample filling is complete. This approach supports the arbitrary patterning and release of different reagents within individual wells without crosstalk for multiplexed analyses. The performance of the in-well spotting technique is characterized using on-chip rolling circle amplification to evaluate its potential for nucleic acid-based diagnostics.
将试剂无缝集成到微流控设备中可显著降低一次性诊断的检测复杂性和成本。在这项工作中，使用可扩展的针点样技术展示了将多重试剂集成到热塑性二维微孔阵列中。使用简单且低成本的窄孔毛细管点样针，可在封闭的纳升级孔阵列中实现高分辨率沉积浓缩试剂。针点样方法还被用于用化学改性蜡层封装沉积的试剂，该蜡层可防止在通过共享微通道引入样品期间干燥的检测成分受到破坏，同时在样品填充完成后实现温度控制释放。这种方法支持在各个孔内对不同试剂进行任意图案化和释放，且无串扰用于多重分析。使用芯片上的滚环扩增来表征孔内点样技术的性能，以评估其在基于核酸的诊断中的潜力。
