【摘要】 数字聚合酶链式反应(dPCR)因高灵敏度、高准确度以及绝对定量的特点，成为分子生物学和临床诊断领域的重要工具，但仍面临集成度不高、样本使用量大及流体控制难等问题。设计了一种集成快速阀门系统和自动加热系统的液滴型dPCR微流控芯片，同时优化了该芯片的制备工艺。研究结果表明，该芯片能够高效、均匀地生成液滴，最大生成频率达5.132 kHz，液滴直径变异系数(CV)值仅为1.67%;形变阀可在0.1 s内快速实现流体通断;集成的片上循环加热系统可实现1.2℃/s的温度升降，温度误差在±0.6℃以内;改进PDMS配方还可有效防止流体蒸发;液滴识别算法准确率高达99.7%。更多还原

【Abstract】 Digital PCR(dPCR) had emerged as a vital tool in molecular biology and clinical diagnostics due to the high sensitivity, accuracy, and absolute quantification capabilities. However, challenges had persisted regarding integration level, sample volume requirements and fluid control. An integrated dropletbased dPCR microfluidic chip incorporating a rapid valve system and automated heating system was presented, along with optimized fabrication processes herein. The results show that the chip may generate droplets efficiently and uniformly, with a maximum generation frequency of 5. 132 kHz and a droplet diameter CV(the coefficient of variation) of only 1. 67%. Rapid fluid control within 0. 1s is enabled by the deformable valve, while temperature ramping rates of 1. 2 ℃/s with temperature variations within ±0. 6 ℃ are achieved by the integrated on-chip thermal cycling system. Fluid evaporation is effectively prevented by an improved PDMS formulation, and the accuracy of 99. 7% is achieved by the droplet recognition algorithm.更多还原