高通量微流控荧光定量PCR激发光非均匀性研究

J4 ›› 2015, Vol. 32 ›› Issue (5): 613-619.

高通量微流控荧光定量PCR激发光非均匀性研究

王华东赵树弥朱灵李志刚王安刘勇

1中国科学院合肥物质科学研究院应用技术研究所安徽省生物医学光学仪器工程技术研究中心，安徽合肥 230031；
2 皖江新兴产业技术发展中心，安徽铜陵 244000

收稿日期:2014-10-30 修回日期:2014-11-25 出版日期:2015-09-28 发布日期:2015-09-28
通讯作者: 王安（1958-）安徽人，研究员，从事光谱学与光谱分析和生物医学光子学方面研究。 E-mail:wangan@aiofm.ac.cn
作者简介:王华东（1990-）安徽人，硕士，从事生物医学光学研究。E-mail：whdm818@163.com
基金资助:
中国科学院战略性先导科技专项基金资助项目（ XDA08040109）

Non-uniform illumination of high-throughput microfluidic qPCR

Wang Huadong, Zhao Shumi, Zhu ling, Li Zhigang, Wang An, Liu Yong

1 Anhui Provincial Engineering Technology Research Center for Biomedical Optical Instrument, Institute of Applied Technology, Hefei Institutes of Physical Science, CAS, Hefei 230031, China;
2 Wanjiang Center for Development of Emerging Industrial Technology, Tongling 244000, China)

Received:2014-10-30 Revised:2014-11-25 Published:2015-09-28 Online:2015-09-28

摘要/Abstract

摘要：

研究高通量微流控荧光定量PCR激发光非均匀性问题对提高DNA浓度定量结果的精度具有重要意义。根据荧光定量PCR的原理，分析了激发光非均匀性对Ct值测量精度的影响，给出了激发光非均匀性引起Ct值测量偏差的表达式，确定了激发光强度标准偏差应小于11.56%的要求。根据微流控芯片结构特点及激发光均匀性要求，设计了基于光棒的匀光光路用于解决激发光照度非均匀性问题。模拟及实验所得到的激发光照度的相对标准偏差分别为3.10%、6.01%，二者均小于11.56%。所设计匀光光路能够满足高通量微流控荧光定量PCR的要求。

关键词: 医用光学与生物技术, 均匀照明, 光学设计, 微流控芯片, 荧光定量PCR

Abstract:

The non-uniform illumination of high-throughput microfluidic qPCR is significant for improving the precision of DNA quantitative results. According to the theoretical principle of qPCR, theoretical analysis is presented, which is focused on the deviation of Ct resulting from non-uniform illumination. And the deviation formula of Ct caused by non-uniform illumination is proposed. Moreover, the requirement of excitation light is determined, that the relative standard deviation of excitation should be less than 11.56%. And then, according to the characteristics of microfluidic chip and the requirement of the excitation light uniformity, an illumination system based on light pipe is designed. The relative standard deviations of irradiance，obtained from simulation and experiment，are respectively 3.10% and 6.01%, both less than 11.56%. And the uniform illumination system is able to meet the requirements of high-throughput microfluidic qPCR.

Key words: medical optics and biotechnology, uniform illumination, optical design, microfluidic chip, qPCR

中图分类号:

O439

王华东赵树弥朱灵李志刚王安刘勇. 高通量微流控荧光定量PCR激发光非均匀性研究[J]. J4, 2015, 32(5): 613-619.

Wang Huadong, Zhao Shumi, Zhu ling, Li Zhigang, Wang An, Liu Yong. Non-uniform illumination of high-throughput microfluidic qPCR[J]. J4, 2015, 32(5): 613-619.

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