CN1629618A

Movatterモバイル変換

Info

Publication number: CN1629618A
Application number: CN 200310122331
Authority: CN
Inventors: 陈义; 黄昊文; 余晓; 齐莉
Original assignee: Institute of Chemistry CAS
Current assignee: Institute of Chemistry CAS
Priority date: 2003-12-16
Filing date: 2003-12-16
Publication date: 2005-06-22

Abstract

本发明是利用表面等离子体共振现象构造的多功能成像仪，主要由P－偏振平行光光源、光入射角调制系统、样品输送与分析系统、反射光纠偏与调制系统、检测器、数据记录与处理系统等六部分组成。该仪器利用P偏振光和棱镜的色散作用激发传感膜表面等离子体的共振而检测共振信号，通过光源的转换可检测到表面等离子体共振成像信号或波长扫描共振信号，对传感膜进行微点阵处理后可达到芯片研究的效果，并能实现成像与共振波长谱图分析转换。它可完成对生物样品和非生物样品组分的定性和定量分析，实现现场或模拟研究分子相互作用的目的，并提供一种快速和非标记的检测方法。本发明仪器结构灵活，可变换和扩展，整个装置简单便宜，易推广使用。

The present invention is a multi-functional imager constructed by using the surface plasmon resonance phenomenon, which mainly consists of a P-polarized parallel light source, a light incident angle modulation system, a sample delivery and analysis system, a reflected light correction and modulation system, a detector, data recording and Processing system and other six parts. The instrument uses P polarized light and the dispersion effect of the prism to excite the resonance of the surface plasmon of the sensing film to detect the resonance signal. The surface plasmon resonance imaging signal or the wavelength scanning resonance signal can be detected through the conversion of the light source, and the sensing film is microscopically detected. After lattice processing, the effect of chip research can be achieved, and the conversion of imaging and resonance wavelength spectrum analysis can be realized. It can complete the qualitative and quantitative analysis of biological samples and non-biological sample components, realize the purpose of on-site or simulated research on molecular interactions, and provide a rapid and non-labeled detection method. The instrument of the invention has a flexible structure, can be changed and expanded, and the whole device is simple and cheap, and easy to popularize and use.

Description

Translated fromChinese

多功能光激发表面等离子体共振成像仪Multi-functional Optically Excited Surface Plasmon Resonance Imager

技术领域technical field

本发明涉及检测技术领域，特别是一种光激发表面等离子体共振仪器系统。具体地说是将光激发的表面等离子体共振信号转变成直观的图像，也可以转换成共振波长探测系统。The invention relates to the technical field of detection, in particular to a light-excited surface plasmon resonance instrument system. Specifically, it converts the light-excited surface plasmon resonance signal into an intuitive image, and can also be converted into a resonance wavelength detection system.

背景技术Background technique

表面等离子体共振(Surface Plasmon Resonance简称SPR)基于P偏振光激发表面等离子体而引起的共振。当P偏振光在高折射介质如玻璃中传播并投射在其界面的自由电子薄层如银或金薄膜时，可在一定角度或波长处形成共振，产生反射衰减，即在某一角度(共振角)或波长(共振波长)处测得反射光强度最小，形成倒峰。共振角或共振波长对紧靠金属膜之介质(即样品)折射率的变化非常敏感，由此可反映样品的组成与性质，实现对样品的实时和无损的检测。原则上，利用SPR原理，能够建立SPR谱图分析、多通道谱图分析和成像分析等方法。近十年来，国际国内对单通道和多通道SPR谱图分析(也称传感)方法研究很多，随着BiocoreAB公司商品仪器的上市，其在生物分子相互作用方面的应用，迅速开展起来，由此反过来推动了仪器设计研究的发展，国内已有关于测定角度(CN1364233A)和波长(CN1203382A)的SPR谱图分析仪器专利，但属单通道范畴。目前最新型仪器是BIAcore 3000(Uppsala，Sweden)，能同时进行四通道测试。为提高测定容量和单位时间的测定速度，美国发展了测定DNA、RNA的相互作用的交叉微通道SPR成像系统并申请了相关专利(PUB.APP.NO.20030017579)，该专利利用硅橡胶制作交叉微通道，但制作过程复杂，实用性较差。美国还有一个最新专利(PUB.APP.NO.20030048452)是关于SPR二维成像的系统，但该专利仍属单功能仪器，即仅涉及二维成像，可扩展性小，不能实现SPR谱图与成像之间的关联或功能转换。Surface Plasmon Resonance (SPR) is based on the resonance caused by the excitation of surface plasmons by P-polarized light. When P-polarized light propagates in a high-refractive medium such as glass and projects on a thin layer of free electrons such as silver or gold film at its interface, resonance can be formed at a certain angle or wavelength, resulting in reflection attenuation, that is, at a certain angle (resonance Angle) or wavelength (resonance wavelength) measured the reflected light intensity minimum, forming an inverted peak. The resonance angle or resonance wavelength is very sensitive to the change of the refractive index of the medium close to the metal film (that is, the sample), which can reflect the composition and properties of the sample, and realize real-time and non-destructive detection of the sample. In principle, using the principle of SPR, methods such as SPR spectrogram analysis, multi-channel spectrogram analysis, and imaging analysis can be established. In the past ten years, there has been a lot of research on single-channel and multi-channel SPR spectrum analysis (also known as sensing) methods at home and abroad. With the listing of commercial instruments from Biocore AB, its application in biomolecular interactions has been rapidly developed. This in turn promotes the development of instrument design and research. There is a patent for SPR spectrogram analysis instrument about measuring angle (CN1364233A) and wavelength (CN1203382A) in China, but it belongs to the category of single channel. At present, the latest instrument is BIAcore 3000 (Uppsala, Sweden), which can perform four-channel testing at the same time. In order to improve the measurement capacity and the measurement speed per unit time, the United States has developed a cross microchannel SPR imaging system for measuring the interaction of DNA and RNA and applied for a related patent (PUB.APP.NO.20030017579), which uses silicon rubber to make cross Microchannel, but the production process is complicated and the practicability is poor. There is also a latest patent in the United States (PUB.APP.NO.20030048452) about the SPR two-dimensional imaging system, but this patent is still a single-function instrument, that is, it only involves two-dimensional imaging, and the scalability is small, and the SPR spectrum cannot be realized. Association or functional transformation between imaging and imaging.

发明内容Contents of the invention

本发明的目的是，提供一种多功能光激发表面等离子体共振成像仪，可以扩展表面等离子体共振成像仪的应用范围及共振信号的相互转变。The object of the present invention is to provide a multi-functional light-excited surface plasmon resonance imager, which can expand the application range of the surface plasmon resonance imager and the mutual conversion of resonance signals.

为达到上述目的，本发明的技术解决方案是提供一种多功能光激发表面等离子体共振成像仪，由P-偏振平行光光源、光入射角调制系统、样品输送与分析系统、反射光纠偏与调制系统、检测器、数据记录与处理系统组成，其在一平台上，设置P-偏振平行光光源，在P-偏振平行光光源射出光的光路上，依序设有光入射角调制系统、样品输送与分析系统、反射光纠偏与调制系统和检测器，检测器与数据记录与处理系统电连接。In order to achieve the above object, the technical solution of the present invention is to provide a multifunctional optically excited surface plasmon resonance imager, which consists of a P-polarized parallel light source, a light incident angle modulation system, a sample delivery and analysis system, reflected light correction and It consists of a modulation system, a detector, and a data recording and processing system. On a platform, a P-polarized parallel light source is set, and on the optical path of the light emitted by the P-polarized parallel light source, a light incident angle modulation system, A sample delivery and analysis system, a reflected light deviation correction and modulation system, and a detector are electrically connected to the data recording and processing system.

所述的多功能光激发表面等离子体共振成像仪，其所述光源，包含连续波长光源，激光器或发光二极管。In the multifunctional photoexcited surface plasmon resonance imager, the light source includes a continuous wavelength light source, a laser or a light emitting diode.

所述的多功能光激发表面等离子体共振成像仪，其所述P-偏振平行光光源，包括激光器、光束扩展器、起偏振器，其中起偏振器可以是晶体或多界面透射—反射结构，理想的起偏振器能输出线偏振光；起偏振器可后置于信号接收结构中。Said multi-functional light-excited surface plasmon resonance imager, said P-polarized parallel light source includes a laser, a beam expander, and a polarizer, wherein the polarizer can be a crystal or a multi-interface transmission-reflection structure, An ideal polarizer outputs linearly polarized light; the polarizer can be placed after the signal receiving structure.

所述的多功能光激发表面等离子体共振成像仪，其所述P-偏振平行光光源，由白光光源、狭缝、单色器、扩束平行光路结构、P-偏振片组成，其中单色器和起偏振器可后置于信号接收系统中。In the multifunctional photoexcited surface plasmon resonance imager, the P-polarized parallel light source is composed of a white light source, a slit, a monochromator, a beam expander parallel optical path structure, and a P-polarizer, wherein the monochromatic The filter and polarizer can be placed in the signal receiving system afterward.

所述的多功能光激发表面等离子体共振成像仪，其用发光二极管为基础光源时，不需要单色器。When the multi-functional light-excited surface plasmon resonance imager uses light-emitting diodes as the basic light source, a monochromator is not needed.

所述的多功能光激发表面等离子体共振成像仪，其所述单色器，为滤光片或光栅。In the multifunctional photoexcited surface plasmon resonance imager, the monochromator is a filter or a grating.

所述的多功能光激发表面等离子体共振成像仪，其所述P-偏振平行光光源的激光器，为单波长或多波长激光器。In the multi-functional light-excited surface plasmon resonance imager, the laser of the P-polarized parallel light source is a single-wavelength or multi-wavelength laser.

所述的多功能光激发表面等离子体共振成像仪，其所述激光器，为He-Ne激光器、染料激光器或半导体激光器。In the multifunctional photoexcited surface plasmon resonance imager, the laser is a He-Ne laser, a dye laser or a semiconductor laser.

所述的多功能光激发表面等离子体共振成像仪，其所述光入射角调制系统，包括入射光臂、信号检测臂、棱镜、传感舌片、精密螺丝杆、电机、角度传感器、导轨；其中，棱镜固定在入射光臂和信号检测臂之间，入射光臂和信号检测臂顶部由螺丝连接，但下面可自由张开或收缩，入射光臂底部连在导轨上；信号检测臂底部与传感舌片相连，由角度传感器控制电机通过精密螺丝杆带动传感舌片在导轨上来回移动，从而使入射光臂和信号检测臂自由张开或收缩达到精确控制棱镜入射光的角度。In the multi-functional light-excited surface plasmon resonance imager, the light incident angle modulation system includes an incident light arm, a signal detection arm, a prism, a sensing tongue, a precision screw, a motor, an angle sensor, and a guide rail; Among them, the prism is fixed between the incident light arm and the signal detection arm, the top of the incident light arm and the signal detection arm are connected by screws, but the bottom can be opened or retracted freely, the bottom of the incident light arm is connected to the guide rail; the bottom of the signal detection arm is connected to the The sensing tongues are connected, and the angle sensor controls the motor to drive the sensing tongue to move back and forth on the guide rail through a precision screw rod, so that the incident light arm and the signal detection arm can freely expand or contract to precisely control the angle of the incident light of the prism.

所述的多功能光激发表面等离子体共振成像仪，其所述光入射角调制系统的调节精确度≤10^-3度。In the multi-functional light-excited surface plasmon resonance imager, the adjustment accuracy of the light incident angle modulation system is ≤10^-3 degrees.

所述的多功能光激发表面等离子体共振成像仪，其所述样品输送与分析系统，由棱镜、传感膜、微升级样品池、进样口、出样口、微量泵及相关管路组成，传感膜为高反射金属膜，棱镜与传感膜用与棱镜折射率相匹配液体介质粘连，传感膜与样品池之间用垫圈密封，样品的输入输出由微量泵控制。In the multi-functional light-excited surface plasmon resonance imager, the sample delivery and analysis system is composed of a prism, a sensing film, a micro-upgraded sample pool, a sample inlet, a sample outlet, a micropump, and related pipelines , the sensing film is a highly reflective metal film, the prism and the sensing film are adhered with a liquid medium that matches the refractive index of the prism, the sensing film and the sample cell are sealed with a gasket, and the input and output of the sample are controlled by a micropump.

所述的多功能光激发表面等离子体共振成像仪，其所述传感膜上，通过控制化学修饰形成定址样品微点阵，用于各种样品分子的识别和检测，该法特别适合于生物大分子及细胞的检测。In the multi-functional light-excited surface plasmon resonance imager, on the sensing film, an addressable sample micro-array is formed through controlled chemical modification, which is used for the identification and detection of various sample molecules. This method is especially suitable for biological Detection of macromolecules and cells.

所述的多功能光激发表面等离子体共振成像仪，其所述反射光纠偏与调制系统，是将反射光经由柱面或球面反射镜和透镜组矫正图像变形，并使图像正确聚焦在检测器上。In the multi-functional light-excited surface plasmon resonance imager, the reflected light deviation correction and modulation system is to correct the image distortion through the reflected light through a cylindrical or spherical mirror and lens group, and make the image correctly focus on the detector superior.

所述的多功能光激发表面等离子体共振成像仪，其所述检测器，为CCD摄像头、CMOS sensor、电荷注入检测器、普通光电管或光电倍增管。In the multi-functional light-excited surface plasmon resonance imager, the detector is a CCD camera, a CMOS sensor, a charge injection detector, an ordinary phototube or a photomultiplier tube.

所述的多功能光激发表面等离子体共振成像仪，其在用连续波长光作光源时，检测器前配接光栅，即转换成测定共振波长的表面等离子体共振仪。The multi-functional light-excited surface plasmon resonance imager, when continuous wavelength light is used as a light source, is equipped with a grating in front of the detector, that is, converted into a surface plasmon resonance imager for measuring resonance wavelengths.

所述的多功能光激发表面等离子体共振成像仪，其所述数据记录与处理系统，设有计算机，检测数据经计算机进行接收、存储、处理并显示或输出。In the multifunctional light-excited surface plasmon resonance imager, the data recording and processing system is provided with a computer, and the detection data is received, stored, processed and displayed or output by the computer.

所述的多功能光激发表面等离子体共振成像仪，其所述数据记录与处理系统，其接收的图像数据，可用商业图形软件离线处理、显示和输出，或可用自编制软件进行实时显示、处理、显示和输出；自编制软件提供全局、局部图像采集的随意选择和实时优化功能，能把表面等离子体共振信号实时输出为一维、二维或三维图像；图像显示可局域任意调控，并同时监测不同样品点信号的动态变化过程；软件具备定性定量功能。In the multifunctional light-excited surface plasmon resonance imager, in the data recording and processing system, the image data received by it can be processed, displayed and output offline by commercial graphics software, or can be displayed and processed in real time by self-programmed software , display and output; the self-compiled software provides free selection and real-time optimization of global and local image acquisition, and can output surface plasmon resonance signals in real time as one-dimensional, two-dimensional or three-dimensional images; image display can be adjusted locally, and Simultaneously monitor the dynamic change process of signals at different sample points; the software has qualitative and quantitative functions.

所述的多功能光激发表面等离子体共振成像仪，其所述金属膜，为金或银膜，厚度在20-80nm之间。In the multi-functional light-excited surface plasmon resonance imager, the metal film is a gold or silver film with a thickness of 20-80nm.

本发明仪器的特点是：The characteristics of the instrument of the present invention are:

1、仪器多功能化：1. Multifunctional instrument:

a)可根据实验需要选择成像方案和图像控制，能实时显示一维、二维和三维图像，图像显示可全局或局域任意选择；a) The imaging scheme and image control can be selected according to the experimental needs, and one-dimensional, two-dimensional and three-dimensional images can be displayed in real time, and the image display can be selected globally or locally;

b)可对各样品点同时进行动态过程分析，形成高通量分析仪器新技术；b) Dynamic process analysis can be performed on each sample point at the same time, forming a new technology of high-throughput analytical instruments;

c)可直接扩展为芯片分析技术；c) It can be directly extended to chip analysis technology;

d)可实现成像与共振波长谱图分析转换。d) The conversion between imaging and resonance wavelength spectrum analysis can be realized.

2、仪器结构灵活、可变换和可扩展：2. The structure of the instrument is flexible, transformable and expandable:

a)光源系统可扩展且灵活，可用激光、白光、发光二极管等来产生P-偏振，偏振或非偏振发光体都可以使用；a) The light source system is scalable and flexible, and P-polarization can be generated by laser, white light, light-emitting diode, etc., and polarized or non-polarized illuminants can be used;

b)图像处理软件可以商业软件，也可以先用本系统特别配置的多功能实时处理软件；b) The image processing software can be commercial software, or the multifunctional real-time processing software specially configured by this system can be used first;

c)可根据实际样品需要变换传感膜，且传感膜的制作方案简单，可采用商业自动点样机，也可以采用自制单头点样器实现对传感膜的点阵化或芯片化处理，包括对样品的定址；c) The sensing film can be changed according to the needs of actual samples, and the production scheme of the sensing film is simple. A commercial automatic pointing machine can be used, or a self-made single-head pointing device can be used to realize the dot matrix or chip processing of the sensing film , including addressing of samples;

d)有多种检测器可共选择，如CCD摄像头、扫描二极管、光电管、光电倍增管、电荷注入检测器及CMOS等。d) A variety of detectors can be selected, such as CCD camera, scanning diode, photoelectric tube, photomultiplier tube, charge injection detector and CMOS.

3、采用光学和机械复合系统来精确的调节光的入射角；3. The optical and mechanical composite system is used to precisely adjust the incident angle of light;

4、分析池结构简单，容易制作，没有微通道结构，适合于传感膜的快速更换。4. The analysis cell has a simple structure, is easy to manufacture, and has no microchannel structure, which is suitable for rapid replacement of the sensing membrane.

5、分析对象广，可用于生物学、生物化学、临床医学和相关的化学及物理检测；可非常方便地进行糖类、DNA、RNA、蛋白质及细胞等相互作用的测定；可执行定性和定量分析。因此可为DNA的杂交及测序、蛋白质组分析以及相关领域的研究提供一种高通量或快速、非标记的检测分析方法。5. It has a wide range of analysis objects and can be used in biology, biochemistry, clinical medicine and related chemical and physical detection; it can be very convenient for the determination of the interaction of sugars, DNA, RNA, proteins and cells; it can perform qualitative and quantitative analyze. Therefore, it can provide a high-throughput or rapid, non-labeled detection and analysis method for DNA hybridization and sequencing, proteome analysis and research in related fields.

附图说明Description of drawings

图1为本发明光激发表面等离子体共振成像仪的基本结构示意图；1 is a schematic diagram of the basic structure of an optically excited surface plasmon resonance imager of the present invention;

图2为本发明光激发表面等离子体共振成像仪的以激光为基础的偏振平行光光源结构示意图；Fig. 2 is a schematic structural diagram of a laser-based polarized parallel light source of an optically excited surface plasmon resonance imager of the present invention;

图3为本发明光激发表面等离子体共振成像仪的光源光束准直结构示意图；Fig. 3 is a schematic diagram of the light source beam collimation structure of the optically excited surface plasmon resonance imager of the present invention;

图4为本发明光激发表面等离子体共振成像仪的光入射角调制系统扩展结构；Fig. 4 is the extended structure of the light incident angle modulation system of the light-excited surface plasmon resonance imager of the present invention;

图5为本发明光激发表面等离子体共振成像仪中样品输送与分析系统扩充结构示意图；Fig. 5 is a schematic diagram of the expanded structure of the sample delivery and analysis system in the optically excited surface plasmon resonance imager of the present invention;

图6为牛血清白蛋白点在本发明成像仪中的共振成像图；Fig. 6 is the resonance imaging diagram of bovine serum albumin spot in the imager of the present invention;

图7为牛血清白蛋白点在本发明成像仪中的共振成像的三维图；Fig. 7 is the three-dimensional diagram of the resonance imaging of the bovine serum albumin spot in the imager of the present invention;

图8为牛血清白蛋白点在本发明成像仪中的共振图的线性叠加图；Fig. 8 is the linear overlay diagram of the resonance diagram of the bovine serum albumin spot in the imager of the present invention;

图9为牛血清白蛋白和聚赖氨酸在本发明成像仪中的共振成像图；Fig. 9 is the resonance imaging diagram of bovine serum albumin and polylysine in the imager of the present invention;

图10为牛血清白蛋白和聚赖氨酸在本发明成像仪中的共振成像的三维图；Figure 10 is a three-dimensional diagram of the resonance imaging of bovine serum albumin and polylysine in the imager of the present invention;

图11为牛血清白蛋白和葡萄糖相互作用在本发明仪器中测定共振波长的表面等离子体共振图；Fig. 11 is the surface plasmon resonance diagram of bovine serum albumin and glucose interaction measuring resonance wavelength in the instrument of the present invention;

图12～图17为本发明的SPR软件流程图。12 to 17 are flowcharts of the SPR software of the present invention.

具体实施方式Detailed ways

本发明为多功能光激发表面等离子体共振成像仪，主要由P-偏振平行光光源A、光入射角调制系统B、样品输送与分析系统C、反射光纠偏与调制系统D、检测器E、数据记录与处理系统F等六部分组成，基本结构如附图1所示。在一平台上，设置P-偏振平行光光源A，在P-偏振平行光光源A射出光的光路上，依序设有光入射角调制系统B、样品输送与分析系统C、反射光纠偏与调制系统D和检测器E，检测器E与数据记录与处理系统F电连接。The present invention is a multifunctional light-excited surface plasmon resonance imager, which mainly consists of a P-polarized parallel light source A, a light incident angle modulation system B, a sample delivery and analysis system C, a reflected light correction and modulation system D, a detector E, The data recording and processing system F is composed of six parts, and the basic structure is shown in Figure 1. On a platform, a P-polarized parallel light source A is set, and on the optical path of the light emitted by the P-polarized parallel light source A, a light incident angle modulation system B, a sample delivery and analysis system C, a reflected light deviation correction system, and The modulation system D and the detector E are electrically connected to the data recording and processing system F.

1.P-偏振平行光光源A：基本结构见附图2包括激光器1、光束扩展器2(由透镜组组成，透镜组包括透镜2a、透镜2b、透镜2c、透镜2d。)、起偏振器3，其中起偏振器3可以是晶体或多界面透射—反射结构，理想的起偏振器3能输出线偏振光。起偏振器3可后置于信号接收结构中。激光器1根据实验所需波长进行更换，通常采用红光半导体激光器。1.P-polarized parallel light source A: basic structure sees accompanying drawing 2 and includes laser 1, beam expander 2 (made up of lens group, lens group includes lens 2a, lens 2b, lens 2c, lens 2d.), polarizer 3. The polarizer 3 can be a crystal or a multi-interface transmission-reflection structure, and an ideal polarizer 3 can output linearly polarized light. The polarizer 3 can be placed downstream in the signal receiving structure. Laser 1 is replaced according to the wavelength required by the experiment, and a red semiconductor laser is usually used.

扩展结构一：以白光为基础的光源，其结构见附图3，由白光光源4、狭缝5、单色器6(如滤光片或光栅等)、扩束平行光路结构7(由7a、7b透镜组组成)、P-偏振片8等组成，其中单色器和起偏振器可后置于信号接收系统中。Extended structure one: a light source based on white light, its structure is shown in Figure 3, consisting of a white light source 4, a slit 5, a monochromator 6 (such as a filter or a grating, etc.), a beam expanding parallel optical path structure 7 (by 7a , 7b lens group), P-polarizer 8, etc., wherein the monochromator and polarizer can be placed in the signal receiving system.

扩展结构二：以发光二极管为基础的光源，不需要单色器，其它结构与白光光源相同。Extended structure 2: A light source based on light-emitting diodes does not require a monochromator, and other structures are the same as the white light source.

扩展结构三：整体光源，分别将各种光源与扩束平行光路结构和起偏振器及单色器设计成整体结构，形成独立的整体光源。Extended structure 3: Integral light source. Various light sources, beam expanders, parallel optical path structures, polarizers and monochromators are designed into an integral structure to form an independent overall light source.

2.光入射角调制系统B：由光学反射部分和机械放大调节部分组成。其中光学反射镜置于机械调节装置上，机械调节装置可实现棱镜入射光角度的精细角度调节，调节精确度达10^-3度或更小。2. Light incidence angle modulation system B: It is composed of an optical reflection part and a mechanical amplification adjustment part. Wherein the optical reflector is placed on the mechanical adjustment device, which can realize the fine angle adjustment of the incident light angle of the prism, and the adjustment accuracy can reach 10^-3 degrees or less.

扩展结构：结构见附图4包括入射光臂9、信号检测臂10、棱镜11、传感舌片12、精密螺丝杆13、电机14、角度传感器15、导轨16。Extended structure: see attached drawing 4 for the structure, including incidentlight arm 9,signal detection arm 10,prism 11, sensingtongue 12,precision screw rod 13,motor 14,angle sensor 15,guide rail 16.

棱镜11固定在入射光臂9和信号检测臂10之间，入射光臂9和信号检测臂10顶部由螺丝连接，但下面可自由张开或收缩，入射光臂9底部连在导轨16上；信号检测臂10底部与传感舌片12相连，由角度传感器控制电机14通过精密螺丝杆13带动传感舌片12在导轨16上来回移动，从而使入射光臂9和信号检测臂10自由张开或收缩达到精确控制棱镜11入射光的角度。Theprism 11 is fixed between the incidentlight arm 9 and thesignal detection arm 10, the top of the incidentlight arm 9 and thesignal detection arm 10 are connected by screws, but the bottom can be freely expanded or contracted, and the bottom of the incidentlight arm 9 is connected to theguide rail 16; The bottom of thesignal detection arm 10 is connected with thesensing tongue 12, and the angle sensor controls themotor 14 to drive the sensingtongue 12 to move back and forth on theguide rail 16 through theprecision screw rod 13, so that the incidentlight arm 9 and thesignal detection arm 10 are opened freely. The angle of the incident light of theprism 11 can be accurately controlled by opening or contracting.

3.样品输送与分析系统C：主要由微量泵和棱镜及相关流路系统组成，样品的流速流量均由微量泵控制，传感膜粘附在棱镜底部，并完成对样品分析。其基本结构见附图5，包括储样瓶17、密封圈18、泵19、样品池20、传感棱镜21、样品流动方向22、废液瓶23。3. Sample delivery and analysis system C: It is mainly composed of a micropump, a prism and related flow path systems. The flow rate and flow rate of the sample are controlled by the micropump, and the sensing film is adhered to the bottom of the prism to complete the sample analysis. Its basic structure is shown in Figure 5, including a storage bottle 17, a sealing ring 18, a pump 19, a sample pool 20, a sensing prism 21, a sample flow direction 22, and a waste liquid bottle 23.

分析系统中的棱镜21通常由光学玻璃制成，也可以采用其它高折射率透光材料制做。传感膜为镀在棱镜表面上的化学修饰金或银膜，金属膜的厚度控制在20-80nm之间。样品由可调节流速的微量泵19引入样品池20，由密封圈18将传感膜和棱镜21接触封接。The prism 21 in the analysis system is usually made of optical glass, and can also be made of other high refractive index light-transmitting materials. The sensing film is a chemically modified gold or silver film plated on the surface of the prism, and the thickness of the metal film is controlled between 20-80nm. The sample is introduced into the sample cell 20 by a micropump 19 with an adjustable flow rate, and the sensing film and the prism 21 are sealed in contact by a sealing ring 18 .

扩展结构：样品输送和清洗系统自动程序控制；传感棱镜21由分立棱镜和传感片组成，中间通过折射率匹配的粘和介质粘接；样品分析池20体可根据需要进行设计和更换。Extended structure: automatic program control of the sample delivery and cleaning system; the sensing prism 21 is composed of a discrete prism and a sensing sheet, and is bonded in the middle by an adhesive with a matching refractive index and a medium; the sample analysis pool 20 can be designed and replaced as required.

4.反射光纠偏与调制系统D：主要由柱透镜或球面反射镜和透镜组成，用于校正因光线通过棱镜后所产生的图像变形和偏差。利用柱透镜可以进行拉长或缩短图像的矫正。通过透镜组的调整，需使矫正后的图像刚好聚焦在CCD检测器E传感表面上。4. Reflected light correction and modulation system D: It is mainly composed of cylindrical lens or spherical mirror and lens, which is used to correct the image deformation and deviation caused by the light passing through the prism. Correction of elongated or shortened images is possible with cylindrical lenses. Through the adjustment of the lens group, the corrected image needs to be just focused on the sensing surface of the CCD detector E.

5.检测器E：根据实验需要选择检测器E及相关配置，在成像结构中，可从CCD摄像头、CMOS、电荷注入检测器E、普通光电管或光电倍增管中选用一种，检测器E可接收从传感膜反射出的全部光束，实时显示共振成像图。5. Detector E: Select the detector E and related configurations according to the needs of the experiment. In the imaging structure, one can be selected from CCD camera, CMOS, charge injection detector E, ordinary phototube or photomultiplier tube. The detector E It can receive all the light beams reflected from the sensing film and display the resonance image in real time.

扩展结构一：检测器E可选用点阵或线阵CCD和扫描光电倍增管，通过快速扫描而成像。Extended structure 1: The detector E can use a dot matrix or linear array CCD and a scanning photomultiplier tube to form an image through fast scanning.

扩展结构二：如需转换成测定共振波长的表面等离子体共振仪，用连续波长光源，并选用与成像结构同样的检测器E，但检测器E前应配接光栅，接收从传感膜反射出的光(亦可用光纤接收并将其传输到光栅中)，由检测器E显示波长扫描共振信号。Extended structure 2: If it needs to be converted into a surface plasmon resonance instrument for measuring the resonance wavelength, use a continuous wavelength light source and use the same detector E as the imaging structure, but a grating should be connected in front of the detector E to receive reflections from the sensing film. The emitted light (which can also be received by an optical fiber and transmitted to the grating), is displayed by the detector E as a wavelength-scanned resonance signal.

6.控制与数据记录处理系统F：该部分主要由计算机和软件组成，完成系统控制及信号记录、处理和输出等任务。图像记录处理所用软件既可以是商业图形软件，也可用自编软件。利用商业软件一般只能采集二维图像，相关数据需离线分析；利用本发明提供的软件可以实现系统控制、实时显示、离线显示、数据后续处理、图形与共振谱线信号选择与转换等操作。自编软件根据实验需要进行设计，可进行二维图像中的同一图像和不同图像的信号比较，实现图像的任意调控并可同时监测多个样品点的动态过程，SPR信号可实时输出为一维、二维及三维图谱且能进行相关谱图的转换，实现定性定量分析。6. Control and data recording and processing system F: This part is mainly composed of computer and software, and completes the tasks of system control and signal recording, processing and output. The software used for image recording and processing can be either commercial graphics software or self-compiled software. Commercial software can generally only collect two-dimensional images, and relevant data needs to be analyzed offline; the software provided by the invention can realize operations such as system control, real-time display, offline display, subsequent data processing, selection and conversion of graphics and resonance spectral line signals. The self-compiled software is designed according to the needs of the experiment. It can compare the signals of the same image and different images in the two-dimensional image, realize the arbitrary regulation of the image and monitor the dynamic process of multiple sample points at the same time. The SPR signal can be output as one-dimensional in real time. , 2D and 3D spectra and can convert related spectra to achieve qualitative and quantitative analysis.

自编软件包括：数据采集、动态分析、静态分析、控制系统和数据存取。数据采集部分是根据实验需要选择适当的采集格式和采集窗口个数，且可以进行图像格式的转换，实现与其它商业软件兼容。动态分析是为了获得SPR的动态信号，该部分可以增强信噪比，能进行单样品点和多样品点的谱线分析，实时地观察不同样品点相互作用的动态过程。静态分析主要是完成定量分析和三维图像处理，获得更加丰富的样品点相互作用的定性定量信息。控制系统部分为了使仪器中各个部分处于最优化的状态，可实现光路系统、样品分析系统的流量、速度和采集控制的调控，能精确地调节光的入射角度，使仪器在最佳条件下工作。数据存取包括数据读取、格式转换和数据保存，实现与其它相关商业软件的兼容。Self-compiled software includes: data acquisition, dynamic analysis, static analysis, control system and data access. The data acquisition part is to select the appropriate acquisition format and the number of acquisition windows according to the needs of the experiment, and can convert the image format to achieve compatibility with other commercial software. Dynamic analysis is to obtain the dynamic signal of SPR. This part can enhance the signal-to-noise ratio, and can perform spectral line analysis of single sample point and multiple sample points, and observe the dynamic process of interaction between different sample points in real time. Static analysis is mainly to complete quantitative analysis and three-dimensional image processing to obtain more qualitative and quantitative information on the interaction of sample points. In order to keep each part of the instrument in an optimal state, the control system can realize the control of the flow, speed and acquisition control of the optical path system and sample analysis system, and can precisely adjust the incident angle of light to make the instrument work under the best conditions. . Data access includes data reading, format conversion and data saving to achieve compatibility with other related commercial software.

全系统工作原理：P偏振平行光A进入光入射角调制系统B，该系统B将光调制到所需角度后进入样品输送与分析系统C，激发棱镜底部传感膜表面等离子体而产生共振，直接导致反射光强度的变化。从棱镜射出的反射光进入反射光纠偏与调制系统D，反射光纠偏与调制系统D矫正图像变形，使矫正后的图像正确聚焦在检测器E上。检测器E电压信号由高速A/D接口如图像采集卡，转换成数字信号，送入数据记录与处理系统F计算机，计算机将所得数据通过软件处理达到定性定量分析目的，并实现图像的实时屏幕显示。The working principle of the whole system: P-polarized parallel light A enters the light incident angle modulation system B, and the system B modulates the light to the required angle and then enters the sample delivery and analysis system C, which excites the surface plasmon of the sensing film at the bottom of the prism to generate resonance. directly lead to changes in the intensity of reflected light. The reflected light emitted from the prism enters the reflected light deviation correction and modulation system D, and the reflected light deviation correction and modulation system D corrects the image distortion, so that the corrected image is correctly focused on the detector E. The voltage signal of the detector E is converted into a digital signal by a high-speed A/D interface such as an image acquisition card, and sent to the computer of the data recording and processing system F. The computer processes the obtained data through software to achieve the purpose of qualitative and quantitative analysis, and realizes the real-time screen of the image show.

实施例一：Embodiment one:

棱镜底部镀有一层50nm厚的金膜作为高反射的传感膜，传感膜首先用巯基十一酸处理，通过化学修饰组装上牛血清白蛋白点。仪器采用CCD作检测器，在溶液中的表面等离子体共振成像图见图6、图7，图8分别为三维图和线性叠加图。The bottom of the prism is coated with a 50nm-thick gold film as a highly reflective sensing film. The sensing film is first treated with mercaptoundecanoic acid, and bovine serum albumin dots are assembled by chemical modification. The instrument uses a CCD as a detector, and the surface plasmon resonance imaging images in solution are shown in Figures 6 and 7, and Figure 8 is a three-dimensional image and a linear overlay image respectively.

实施例二：Embodiment two:

与实施例一用相同的方法修饰传感膜，分别修饰牛血清白蛋白和聚赖氨酸。仪器采用CCD作检测器，在溶液中的表面等离子体共振成像图见图9(左三列为牛血清白蛋白，右三列为聚赖氨酸)，图10为其三维图。The same method was used to modify the sensing membrane as in Example 1, respectively modifying bovine serum albumin and polylysine. The instrument uses a CCD as a detector, and the surface plasmon resonance image in the solution is shown in Figure 9 (the third column on the left is bovine serum albumin, and the third column on the right is polylysine), and Figure 10 is its three-dimensional view.

实施例三：Embodiment three:

固定入射光角度采用CCD作检测器并配接光栅，与实施例一用相似的方法修饰传感膜，传感膜上接有牛血清白蛋白。首先往样品池通入缓冲溶液，然后再通入配于相同缓冲溶液中的葡萄糖样品，同时启动实时检测操作，分别记录缓冲溶液和葡萄糖溶液共振波长的变化，见图11，由此实现对葡萄糖的实时与动态分析。A CCD was used as a detector and a grating was used to fix the angle of incident light, and a similar method was used to modify the sensing film as in Example 1, and the sensing film was connected with bovine serum albumin. First, the buffer solution is passed into the sample pool, and then the glucose sample prepared in the same buffer solution is passed through, and the real-time detection operation is started at the same time, and the changes in the resonance wavelength of the buffer solution and the glucose solution are recorded respectively, as shown in Figure 11, thereby realizing the detection of glucose real-time and dynamic analysis.

本发明的SPR软件流程，请见图12～图17。The SPR software flow of the present invention is shown in Figs. 12-17.