School of Mechanical and Manufacturing Engineering, Dublin City University, Glasnevin, Dublin, Ireland; National Centre for Sensor Research (NCSR), Dublin City University, Dublin, Ireland; Biodesign Europe, Dublin City University, Dublin, Ireland.
School of Mechanical and Manufacturing Engineering, Dublin City University, Glasnevin, Dublin, Ireland.
Anal Chim Acta. 2023 Nov 1;1280:341859. doi: 10.1016/j.aca.2023.341859. Epub 2023 Sep 30.
Lab-on-a-disc (LoaD) technology has emerged as a transformative approach for point-of-care diagnostics and high-throughput testing. The promise of integrating multiple laboratory functions onto a single integrated platform has significant implications for healthcare, especially in resource-limited settings. However, one of the primary challenges faced in the design and manufacture of LoaD devices is the integration of effective valving mechanisms. These valves are essential for fluid control and routing, but their intricacy often leads to complexities in design and increased vulnerability to failure. This emphasizes the need for improved designs and manufacturing processes without complex, integrated valving mechanisms. (96) RESULTS: We describe a fully automated biological workflow and reagent actuation on a LoaD device without an integrated valving system. The Two Degrees-of-Freedom (2DoF) custom centrifuge alters the centre of rotation, facilitating fluid flow direction changes on the microfluidic platform through a custom programmed interface. A novel 360-degree fluid manipulation approach via secondary planetary gear motion enabled sequential assay reagent actuation without embedded valve triggering, with the addition of infinite incubation times and efficient use of platform realty. The simplified LoaD platform uses clever design, with intermediate flow chambers to avoid cross contamination between reagent steps. Notably, the optimized LoaD platform demonstrated a two-fold DNA yield at higher HEK-293 cell concentrations compared to commercially available spin-column kits. This significantly simplified LoaD platform successfully automated a common, complex workflow without inhibiting DNA purification. (129) SIGNIFICANCE: This system exhibits the clever coupling of both 2DoF and centrifugal microfluidics to create an autonomous testing package capable of eradicating the need for complex valving systems to automate biological workflows on LoaDs. This automated system has outperformed commercially available DNA extraction kits for higher cell counts. The platform's elimination of valve requirements ensures unlimited sample incubation times and enhances reliability, making it a straightforward option for automated biological workflows, particularly in diagnostics. (73).
碟式实验室 (LoaD) 技术已成为即时诊断和高通量检测的变革性方法。将多个实验室功能集成到单个集成平台上的承诺对医疗保健具有重要意义，特别是在资源有限的环境中。然而，在设计和制造 LoaD 设备时面临的主要挑战之一是有效阀门机制的集成。这些阀门对于流体控制和路由至关重要，但它们的复杂性常常导致设计复杂，并增加故障的脆弱性。这强调了需要改进设计和制造工艺，而无需复杂的集成阀门机制。（96）结果：我们描述了在没有集成阀系统的 LoaD 设备上的全自动生物工作流程和试剂驱动。双自由度 (2DoF) 定制离心机改变了旋转中心，通过定制编程接口在微流控平台上改变流体流动方向。通过二次行星齿轮运动实现的新颖 360 度流体操纵方法，无需嵌入式阀触发，即可实现连续的分析试剂驱动，同时增加了无限的孵育时间并有效利用平台实际情况。简化的 LoaD 平台采用巧妙的设计，使用中间流室避免试剂步骤之间的交叉污染。值得注意的是，与市售的 spin-column 试剂盒相比，优化的 LoaD 平台在更高的 HEK-293 细胞浓度下显示出两倍的 DNA 产量。这个简化的 LoaD 平台成功地在没有抑制 DNA 纯化的情况下自动执行了一个常见的复杂工作流程。（129）意义：该系统展示了双 2DoF 和离心微流控的巧妙结合，创建了一个自主测试包，能够消除在 LoaD 上自动化生物工作流程对复杂阀门系统的需求。这个自动化系统在更高的细胞计数下优于市售的 DNA 提取试剂盒。该平台消除了对阀门的需求，确保了无限的样品孵育时间，并提高了可靠性，使其成为自动化生物工作流程的简单选择，特别是在诊断领域。（73）。
