高等学校化学学报

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单波长荧光交叉相关光谱单分子检测系统

谢超, 董朝青, 任吉存   

  1. 上海交通大学化学化工学院, 上海 200240
  • 收稿日期:2007-11-06 修回日期:1900-01-01 出版日期:2008-05-10 发布日期:2008-05-10
  • 通讯作者: 任吉存

Fluorescence Cross-correlation Spectroscopy Using Single Wavelength Laser Excitation

XIE Chao, DONG Chao-Qing, REN Ji-Cun*   

  1. College of Chemistry and Chemical Engineering, Shanghai Jiaotong University, Shanghai 200240, China
  • Received:2007-11-06 Revised:1900-01-01 Online:2008-05-10 Published:2008-05-10
  • Contact: REN Ji-Cun

摘要: 基于激光共焦构型建立了一套单波长荧光交叉相关光谱检测系统, 并对检测系统进行了优化, 阐述了荧光交叉(相关)光谱基本理论. 以荧光量子点和有机染料为标记探针, 以人免疫球蛋白与羊抗人免疫球蛋白的结合反应为研究对象, 利用该系统成功地实现了在单分子水平上对免疫结合产物的分子数浓度、特征扩散时间和动力学半径等参数的表征.

关键词: 荧光交叉相关光谱, 单分子检测, 单波长激发, 量子点

Abstract: The basic principle of fluorescence cross-correlation spectroscopy were introduced in this article. A setup of fluorescence cross-correlation spectroscopy(FCCS) was established with single wavelength laser. After the optimization of the setup, the detection volume is about 0.7 fL. This home-built setup was successfully applied for study of the binding reaction of human immunoglobulin G with goat anti-human immunoglobulin G. With quantum dots(745 nm) and Rhodamine B(580 nm) as probes and 532 nm laser beam as excitation source, the cross-talk effect was near completely suppressed. The molecule numbers in a highly focused volume, concentration, diffusion time and hydrodynamic radius of the reaction product can be determined by FCCS system.

Key words: Fluorescence cross-correlation spectroscopy, Single-molecule detection, Single wavelength laser excitation, Quantum dot

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