表面等离子体子共振成像系统的研制和应用
发布时间：2018-08-22 09:53
【摘要】：表面等离子体子共振(Surface Plasmon Resonance, SPR)成像技术被广泛应用于生物分子相互作用的研究。本技术具有实时、无标记、灵敏度高的特点,使其在提供分子相互作用的动态信息上有巨大的优势。相应地,研制高通量、高精度、精简实用的SPR检测系统,是实现SPR技术优势的关键。本论文从多个方面改进了SPR检测系统的性能,具体包括： 1、采用新颖的角度扫描结构,研制了一台高分辨率的角度扫描SPR成像系统。本系统的扫描检测平台采用压电陶瓷直线电机与四边形的传动装置的结构,其扫描角度的分辨率达10-4度。同时开发操作控制界面,用于操控电机、相机等电子元件,并进行数据处理分析。整个系统最终的重复精度为1×10-3度。 2、使用本SPR成像系统高通量的检测caspase-3基因的碱基错配：使用自制的点样仪制作DNA探针阵列,并与目标DNA(caspase-3)杂交；结果显示使用本系统可同时检测多个不同碱基错配的目标DNA,其检出限达25pmol/L。使用该系统观测癌细胞：在原仪器装置的基础上,组装了一台双视场的SPR细胞成像装置,在正常视场和SPR视场下对HepG2癌细胞同时成像；两个视场的综合比较可以排除许多外界干扰,并获得癌细胞形态的信息。 3、开发了一种适用于SPR检测的无动力流体注射系统。本进样系统利用液体自身的重力实现连续的进样。本重力引导的流体注射系统可提供相对稳定的进样速度,仅需要简单的手动操作。并在研究互补DNA单链杂交的过程中,成功地提取出杂交过程的动态信息。最后,组装了一台手持的小型SPR成像装置以用于现场检测。
[Abstract]:Surface plasmon resonance (Surface Plasmon Resonance, SPR) imaging is widely used in the study of biomolecular interactions. This technique has the characteristics of real-time, unlabeled and high sensitivity, which makes it have a great advantage in providing dynamic information of molecular interaction. Accordingly, the development of high-throughput, high-precision, simplified and practical SPR detection system is the key to realize the advantages of SPR technology. In this paper, the performance of SPR detection system is improved from many aspects, including: 1. A high resolution angle scanning SPR imaging system is developed using a novel angle scanning structure. The scanning and detecting platform of this system adopts the structure of piezoelectric ceramic linear motor and quadrilateral transmission, and its scanning angle resolution is 10 ~ (-4) degrees. At the same time, the operating control interface is developed, used to control the motor, camera and other electronic components, and data processing and analysis. The final repetition accuracy of the whole system is 1 脳 10 ~ (-3) degrees. The SPR imaging system is used to detect the base mismatch of caspase-3 gene in high throughput. The DNA probe array is made by using a self-made spotting instrument and hybridized with the target DNA (caspase-3). The results show that the system can simultaneously detect multiple target DNAs with different base mismatches, and the detection limit is up to 25pmol / L. Using this system to observe cancer cells: on the basis of the original instrument, a dual-field SPR cell imaging device was assembled, and the HepG2 cancer cells were simultaneously imaged under the normal field of view and the SPR field of view. The comprehensive comparison of the two fields of view can eliminate many external interference and obtain morphological information of cancer cells. 3. A non-dynamic fluid injection system for SPR detection is developed. The system uses the gravity of the liquid itself to realize continuous injection. This gravity-guided fluid injection system provides a relatively stable injection rate, requiring only simple manual operation. The dynamic information of the hybridization process was successfully extracted during the study of complementary DNA single strand hybridization. Finally, a small hand-held SPR imaging device is assembled for field detection.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R310
本文编号：2196745
[Abstract]:Surface plasmon resonance (Surface Plasmon Resonance, SPR) imaging is widely used in the study of biomolecular interactions. This technique has the characteristics of real-time, unlabeled and high sensitivity, which makes it have a great advantage in providing dynamic information of molecular interaction. Accordingly, the development of high-throughput, high-precision, simplified and practical SPR detection system is the key to realize the advantages of SPR technology. In this paper, the performance of SPR detection system is improved from many aspects, including: 1. A high resolution angle scanning SPR imaging system is developed using a novel angle scanning structure. The scanning and detecting platform of this system adopts the structure of piezoelectric ceramic linear motor and quadrilateral transmission, and its scanning angle resolution is 10 ~ (-4) degrees. At the same time, the operating control interface is developed, used to control the motor, camera and other electronic components, and data processing and analysis. The final repetition accuracy of the whole system is 1 脳 10 ~ (-3) degrees. The SPR imaging system is used to detect the base mismatch of caspase-3 gene in high throughput. The DNA probe array is made by using a self-made spotting instrument and hybridized with the target DNA (caspase-3). The results show that the system can simultaneously detect multiple target DNAs with different base mismatches, and the detection limit is up to 25pmol / L. Using this system to observe cancer cells: on the basis of the original instrument, a dual-field SPR cell imaging device was assembled, and the HepG2 cancer cells were simultaneously imaged under the normal field of view and the SPR field of view. The comprehensive comparison of the two fields of view can eliminate many external interference and obtain morphological information of cancer cells. 3. A non-dynamic fluid injection system for SPR detection is developed. The system uses the gravity of the liquid itself to realize continuous injection. This gravity-guided fluid injection system provides a relatively stable injection rate, requiring only simple manual operation. The dynamic information of the hybridization process was successfully extracted during the study of complementary DNA single strand hybridization. Finally, a small hand-held SPR imaging device is assembled for field detection.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R310
【参考文献】
相关期刊论文 前7条
1 赵晓君,金钦汉;光纤表面等离子体共振生物探针的研究[J];抚顺石油学院学报;1996年03期
2 赵晓君,陈焕文,宋大千,牟颖,张寒琦,金钦汉;表面等离子体子共振传感器I:基本原理[J];分析仪器;2000年04期
3 赵晓君,王珍,许汉英,梁枫,张寒琦,金钦汉;表面等离子体激元共振光化学传感器的研究[J];高等学校化学学报;1998年08期
4 赵晓君,牟颖,王珍,张寒琦,金钦汉,许汉英;基于表面等离子体子共振的白蛋白免疫传感器的研究[J];高等学校化学学报;1999年05期
5 牟颖,赵晓君,王珍,张寒琦,金钦汉;γ-干扰素DNA传感器组装过程的表面等离子体子共振研究[J];化学学报;2000年05期
6 崔大付,李向明,蔡浩原,李亚亭,郑自攀,陈翔;表面等离子体谐振(SPR)生化分析仪的研制[J];现代科学仪器;2001年06期
7 赵杰,王景田,韩泾鸿;SPR生物化学量检测系统[J];仪表技术与传感器;1996年03期
，本文编号：2196745
本文链接：https://www.wllwen.com/yixuelunwen/swyx/2196745.html
