化学
荧光
微流控
共焦
微流控芯片
污染
炸薯条
激光器
分析化学(期刊)
纳米技术
光学
色谱法
材料科学
物理
工程类
电气工程
生物
生态学
作者
Xiurui Zhu,Baoxia Liu,Shisheng Su,Bo Wang,Yu Bai,Haiwang Huang,Xiaobin Liu,Xin Cheng,Xianhua Wang,Lingxiang Zhu,Wei Yang,Na Gao,Gaoshan Jing,Yong Guo
出处
期刊:Talanta
[Elsevier]
日期:2020-01-01
卷期号:206: 120200-120200
被引量:9
标识
DOI:10.1016/j.talanta.2019.120200
摘要
Highly-sensitive and contamination-free droplet digital PCR (ddPCR) is an enabling technology and widely needed for accurate quantification of nucleic acid in clinical applications. In this paper, a novel droplet reader was developed by combining a "quasi" confocal laser-induced fluorescence (LIF) cytometry with a delicate microfluidic chip design. The droplets with a size of 90 μm was illuminated at an out-of-focus position by two aligned laser beams to generate maximum fluorescent signal. Additionally, the lateral offset position of the microfluidic chip should be precisely tuned so that the bandwidth of the FAM and VIC channels were configured at the matching sizes. Then, PMT gain voltages and pneumatic pressures were optimized for better droplet detection efficiencies. An aerosol adsorption experiment was performed to demonstrate that there was no aerosol contamination, and detected copy numbers of both mutants and wild types scaled linearly with the expected input copy numbers (r2>0.998) with a LoB of 0.0 copies and LoD of 3.0 copies. The results demonstrated that this droplet reader with the delicate chip is a convenient, highly-sensitive and contamination-free to detect fluorescence signals inside droplets after ddPCR, which is highly promising for broad applications of ddPCR in clinical diagnosis.
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