Improved picoliter-sized micro-reactors for high-throughput biological analysis  

作　　者：HAN WeiJing YUAN LiNa WEI QingQuan LI YunTao REN LuFeng ZHOU XiaoGuang YU Jun YU YuDe 

机构地区：[1]State Key Laboratory of Integrated Optoelectronics,Institute of Semiconductors,Chinese Academy of Sciences [2]The Joint Laboratory of Bioinformation Acquisition and Sensing Technology,Institute of Semiconductors,Beijing Institute of Genomics,Chinese Academy of Sciences [3]DNA Sequencing Technologies R&D Center,Beijing Institute of Genomics,Chinese Academy of Sciences

出　　处：《Science China(Life Sciences)》2013年第12期1134-1141,共8页中国科学（生命科学英文版）

基　　金：supported by the Scientific Equipment Research Project of Chinese Academy of Sciences(YZ200823)

摘　　要：High-throughput pyrosequencing,carried out in millions of picoliter-sized reactors on a fiber-optic slide,is known for its longer read length.However,both optical crosstalk(which reduces the signal-to-noise ratio of CCD images)and chemical retention adversely affect the accuracy of chemiluminescence determination,and ultimately decrease the read length and the accuracy of pyrosequencing results.In this study,both titanium and oxidized aluminum films were deposited on the side walls and upper faces of micro-reactor slides to enhance optical isolation;the films reduced the inter-well crosstalk by one order of magnitude.Subsequently,chemical retention was shown to be caused by the lower diffusion coefficient of the side walls of the picolitersized reactors because of surface roughness and random pores.Optically isolated fiber-optic slides over-coated with silicon oxide showed smoother surface morphology,resulting in little chemical retention;this was further confirmed with theoretical calculations.Picoliter-sized micro-reactors coated with titanium-silicon oxide films showed the least inter-well optical crosstalk and chemical retention;these properties are expected to greatly improve the high-throughput pyrosequencing performance.High-throughput pyrosequencing, carried out in millions of picoliter-sized reactors on a fiber-optic slide, is known for its long- er read length. However, both optical crosstalk （which reduces the signal-to-noise ratio of CCD images） and chemical retention adversely affect the accuracy of chemiluminescence determination, and ultimately decrease the read length and the accuracy of pyrosequencing results. In this study, both titanium and oxidized aluminum films were deposited on the side walls and upper faces of micro-reactor slides to enhance optical isolation; the films reduced the inter-well crosstalk by one order of magnitude. Subsequently, chemical retention was shown to be caused by the lower diffusion coefficient of the side walls of the picoliter- sized reactors because of surface roughness and random pores. Optically isolated fiber-optic slides over-coated with silicon oxide showed smoother surface morphology, resulting in little chemical retention; this was further confirmed with theoretical calculations. Picoliter-sized micro-reactors coated with titanium-silicon oxide films showed the least inter-well optical cross-talk and chemical retention; these properties are expected to greatly improve the high-throughput pyrosequencing performance.

关 键 词：high throughput analysis picoliter-sized micro-reactor surface coating 

分 类 号：Q503[生物学—生物化学]