高等学校化学学报

高等学校化学学报

• 研究快报 • 上一篇    下一篇

单分子流式检测仪的研制

杨玲玲, 朱少彬, 杭纬, 颜晓梅   

  1. 厦门大学化学化工学院化学生物学系, 化学生物学福建省重点实验室, 现代分析科学教育部重点实验室, 厦门 361005
  • 收稿日期:2008-03-17 修回日期:1900-01-01 出版日期:2008-08-10 发布日期:2008-08-10
  • 通讯作者: 颜晓梅

Development of a Single-molecule Flow Analyzer

YANG Ling-Ling, ZHU Shao-Bin, HANG Wei, YAN Xiao-Mei   

  1. Department of Chemical Biology, College of Chemistry and Chemical Engineering, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Sciences of Ministry of Education, Xiamen University, Xiamen 361005, China
  • Received:2008-03-17 Revised:1900-01-01 Online:2008-08-10 Published:2008-08-10
  • Contact: YAN Xiao-Mei

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被引次数

13

摘要： 采用激光诱导荧光和流体动力学聚焦技术成功地研制出单分子流式检测仪, 实现了对水溶液中单个藻红蛋白及单个DNA分子片段的检测, 检测速率可达到每秒几十次. 与单分子荧光显微术相比, 流式分析将固定的标本台改为流动的单分子悬液, 大大提高了检测速率和统计精确性, 更加适合生物样品的快速、超高灵敏分析.

关键词: 单分子检测, 流体聚焦, 藻红蛋白

Abstract: A single-molecule flow analyzer based on laser induced fluorescence and hydrodynamic focusing was developed in our lab. There were three key aspects of this design for achieving single molecule detection sensitivity: (1) a detector with a high photon-detection efficiency(>65% for avalanche photo-diode detector); (2) a small probe volume obtained by sheath-protected, hydrodynamically focused sample stream to reduce background; and (3) a slow flow to provide extended dwell time(milliseconds) in the probe volume so that more fluorescence cycling can occur. The single molecule detection sensitivity was demonstrated by the successful detection of single phycoerythrin molecules. The house-constructed single-molecule flow analyzer was applied to DNA fragment sizing with linear correlation between DNA fragment length and mean burst size.

Key words: Single-molecule detection, Hydrodynamic focusing, Phycoerythrin

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