液滴数字PCR芯片结果自动化读出平台研究.docVIP

液滴数字PCR芯片结果自动化读出平台研究 　　摘 要： 液滴数字聚合酶链式反应（ddPCR）作为第三代PCR技术，可实现核酸分子绝对定量。然而，在数字PCR结果读出过程中，对1 000万量级的液滴进行统计费时费力而且容易出错。针对此情况设计了液滴数字PCR芯片的自动扫描识别平台，系统包括硬件移动平台和软件拍照、拼接和识别功能。软件控制平移台顺序移动并控制摄像头拍照，移动完成后把拍得的照片拼接成一张图片，之后对图片中的液滴进行识别。此平台通过二维平移台的精确定位扫描及图像分割、识别算法，对荧光和非荧光液滴识别率达到99.6%以上。平台已经应用到了实验室中，对数字PCR芯片上的微液滴可实现一键识别，极大的减小了识别时间和错误率。 　　关键词： ddPCR； 核酸分子定量； 自动扫描； 液滴识别 　　中图分类号： TN911?34 文献标识码： A 文章编号： 1004?373X（2017）18?0001?06 　　Research on automatic result readout platform of droplet digital PCR chip 　　LIU Songsheng1， 2， YUAN Haojun2， LIU Qiang1， 2， ZHOU Hongbo2， MAO Hongju2， REN Wei1 　　（1. International Center of Quantum and Molecular Structures， Shanghai University， Shanghai 201900， China； 　　2. Shanghai Institute of Microsystem and Information Technology， Chinese Academy of Sciences， Shanghai 200050， China） 　　Abstract： As the third generation PCR technique， the droplet digital PCR （ddPCR） can realize the absolute quantification for nucleic acid molecule， but in the process of PCR result readout， the statistics of ten?million magnitude droplets wastes time and manpower， and is easy to result in errors. Therefore， an automatic scanning and recognition platform of ddPCR chip was designed. The system includes the hardware mobile platform， and has the functions of software photographing， image splicing and droplet recognition. The sequential movement of the platform and photographing of camera are controlled by the software， and then the photographed pictures are spliced into a photograph to recognize droplets in it. The recognition rate of the platform for fluorescent and non?fluorescent droplets can reach up to above 99.6% by means of the accurate positioning and scanning， image segmentation and recognition algorithms of the 2D translation platform. This platform has been applied to the laboratory， can recognize the microdroplets produced by digital PCR chip with one key， shorten the recognition time and reduce the recognition error rate greatly. 　　Keywords： ddPCR； nucleic acid