CN103115901B - Device for detecting biological chips based on resonance light scattering - Google Patents
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Abstract
基于共振光散射检测生物芯片的装置，包括光源驱动器、光纤、激发光源、耦合透镜组、生物芯片载物台、光电传感器、上位机、运动平台和多轴电机驱动；光源驱动器控制激发光源产生激发光，激发光经光纤输出至耦合透镜组并对其进行光束整形并入射至生物芯片载物台上，以入射角大于临界角由生物芯片侧边入射，样品受到激发光的激发后发生共振光散射，向外辐射特定波长的共振散射光，共振散射光入射至光电传感器中，上位机接收转换后的电信号并通过上位机软件对其进行采集分析，得到扫描图像，上位机软件控制多轴电机驱动运行并驱动运动平台移动，带动生物芯片载物台移动，对生物芯片的整个点样区域进行扫描。本发明背景噪声低，检测灵敏度高。
A device for detecting biochips based on resonance light scattering, including a light source driver, an optical fiber, an excitation light source, a coupling lens group, a biochip stage, a photoelectric sensor, a host computer, a motion platform, and a multi-axis motor drive; the light source driver controls the excitation light source to generate excitation Light, the excitation light is output to the coupling lens group through the optical fiber and the beam is shaped and incident on the biochip stage. The incident angle is greater than the critical angle and is incident from the side of the biochip. The sample is excited by the excitation light and generates resonance light. Scattering, the resonant scattered light of a specific wavelength is radiated outward, and the resonant scattered light is incident on the photoelectric sensor. The host computer receives the converted electrical signal and collects and analyzes it through the host computer software to obtain a scanning image. The host computer software controls the multi-axis The motor is driven to run and drive the motion platform to move, which drives the biochip carrier to move, and scans the entire sampling area of the biochip. The invention has low background noise and high detection sensitivity.
Description
技术领域technical field
本发明涉及生物芯片检测技术领域，具体涉及一种基于共振光散射检测生物芯片的装置。The invention relates to the technical field of biochip detection, in particular to a device for detecting biochips based on resonance light scattering.
背景技术Background technique
生物芯片（Microarray）是近几年来在生命科学领域中迅速发展起来的一项高新技术，它主要是指通过微加工技术在只有几平方厘米的固体芯片表面构建的微生物化学系统，以实现对DNA、蛋白质、细胞等生物样品的快速、准确与高通量的检测，其主要原理是采用化学或物理方法，将大量探针固化于支持物的表面，然后根据生物分子之间的亲和作用，如核酸分子的碱基配对作用、抗原抗体的结合等进行反应，再对反应信号进行检测分析，即可得到该样品的相关信息。由于该技术可以将大量的探针同时固定于支持物上，所以一次就可以对大量的生物分子进行检测分析，具有高自动化、高通量的特点，随着生物芯片技术的发展，对生物芯片检测技术也提出了更高的要求。Microarray is a high-tech that has developed rapidly in the field of life sciences in recent years. It mainly refers to the microbial chemical system constructed on the surface of a solid chip with only a few square centimeters through micro-processing technology to realize DNA Rapid, accurate and high-throughput detection of biological samples such as proteins, cells, etc. The main principle is to use chemical or physical methods to immobilize a large number of probes on the surface of the support, and then according to the affinity between biomolecules, Such as the base pairing of nucleic acid molecules, the combination of antigen and antibody, etc., and then detect and analyze the reaction signal, the relevant information of the sample can be obtained. Because this technology can immobilize a large number of probes on the support at the same time, it can detect and analyze a large number of biomolecules at one time, which has the characteristics of high automation and high throughput. With the development of biochip technology, biochips Detection technology also puts forward higher requirements.
目前，对于生物芯片的检测主要有两种方式，一种方式是通过荧光方法进行检测，这种检测方式需要荧光分子对生物芯片进行标记，由于荧光分子本身亮度有限，容易发生光淬灭和光漂白现象，对检测结果影响较大；另一种检测方式是通过生物芯片扫描仪对生物芯片进行检测，这种检测方式存在着背景噪声较高、检测灵敏度偏低的缺陷，对检测结果产生严重的影响，因此研发具有高灵敏度、高分辨率的生物芯片检测装置对于发展生物芯片技术具有重要意义。At present, there are two main ways to detect biochips. One way is to detect by fluorescence method. This detection method requires fluorescent molecules to label biochips. Due to the limited brightness of fluorescent molecules themselves, photoquenching and photobleaching are prone to occur. The phenomenon has a great impact on the detection results; another detection method is to detect the biochip through a biochip scanner. This detection method has the defects of high background noise and low detection sensitivity, which has a serious impact on the detection results. Therefore, the development of biochip detection devices with high sensitivity and high resolution is of great significance for the development of biochip technology.
发明内容Contents of the invention
为了解决现有生物芯片扫描仪检测生物芯片存在的背景噪音过高和灵敏度偏低的问题，本发明提供一种基于共振光散射检测生物芯片的装置。In order to solve the problems of high background noise and low sensitivity in detecting biochips in existing biochip scanners, the present invention provides a device for detecting biochips based on resonance light scattering.
本发明为解决技术问题所采用的技术方案如下：The technical scheme that the present invention adopts for solving technical problems is as follows:
基于共振光散射检测生物芯片的装置，该装置包括光源驱动器、光纤、激发光源、耦合透镜组、生物芯片载物台、光电传感器、上位机、运动平台和多轴电机驱动；A device for detecting biochips based on resonance light scattering, which includes a light source driver, an optical fiber, an excitation light source, a coupling lens group, a biochip stage, a photoelectric sensor, a host computer, a motion platform, and a multi-axis motor drive;
所述光源驱动器控制激发光源产生激发光，激发光通过光纤输出至耦合透镜组并对其进行光束整形，整形后的激发光入射至生物芯片载物台上，激发光以入射角大于临界角的角度由生物芯片侧边入射，生物芯片上的样品受到激发光的激发后发生共振光散射，向外辐射特定波长的共振散射光，共振散射光入射至光电传感器中，光信号转换为电信号，上位机接收转换后的电信号并通过上位机中的上位机软件对其进行采集分析，得到扫描图像，上位机软件控制多轴电机驱动运行并驱动运动平台移动，同时带动生物芯片载物台移动，对生物芯片的整个点样区域进行扫描。The light source driver controls the excitation light source to generate excitation light, the excitation light is output to the coupling lens group through the optical fiber and the beam is shaped, the shaped excitation light is incident on the biochip stage, and the excitation light has an incident angle greater than the critical angle. The angle is incident from the side of the biochip, and the sample on the biochip is excited by the excitation light and undergoes resonant light scattering, and radiates resonant scattered light of a specific wavelength outward. The resonant scattered light is incident on the photoelectric sensor, and the optical signal is converted into an electrical signal. The upper computer receives the converted electrical signal and collects and analyzes it through the upper computer software in the upper computer to obtain the scanned image. The upper computer software controls the multi-axis motor to drive and drive the motion platform to move, and at the same time drives the biochip stage to move , to scan the entire spotting area of the biochip.
所述运动平台主要由三个直线位移平台组成。The motion platform is mainly composed of three linear displacement platforms.
所述生物芯片表面放置有样品，样品受到激发光的激发后发生共振光散射，向外辐射特定波长的散射光。A sample is placed on the surface of the biochip, and the sample undergoes resonant light scattering after being excited by the excitation light, and radiates scattered light of a specific wavelength outward.
所述上位机软件包括：图像采集、扫描控制、相机控制、导轨控制、扫描数据分析和数据复位控制。The host computer software includes: image acquisition, scanning control, camera control, rail control, scanning data analysis and data reset control.
发明原理：本发明是基于共振光散射检测原理来检测生物芯片，共振光散射（Resonance Light-Scattering，RLS）是一种具有极高灵敏度和选择性的研究方法，它利用了金属纳米粒子在白光激发光源中会产生强烈的散射光的原理，即当激发光波长落在分子的吸收带时，分子中的电子由于能级跃迁作用，向所有方向辐射出极强的散射光，这种现象被称为共振光散射。Invention Principle: The present invention is based on the detection principle of resonance light-scattering to detect biochips. Resonance Light-Scattering (RLS) is a research method with extremely high sensitivity and selectivity. The principle of strong scattered light in the excitation light source, that is, when the wavelength of the excitation light falls on the absorption band of the molecule, the electrons in the molecule radiate extremely strong scattered light in all directions due to the energy level transition. This phenomenon is called called resonance light scattering.
共振光散射检测是一种暗场检测方式，激发光源光线不直接进入检测器视场，检测器收集的光子仅来自于金属纳米粒子的散射光，噪声来自于检测器的暗电流噪声，因此可以有效地提高信噪比和检测的灵敏度。同时，共振光散射检测技术使用白光作为检测光源，设备简单，价格低廉，检测成本大幅下降。Resonant light scattering detection is a dark field detection method. The light from the excitation light source does not directly enter the field of view of the detector. The photons collected by the detector only come from the scattered light of metal nanoparticles, and the noise comes from the dark current noise of the detector. Therefore, it can Effectively improve the signal-to-noise ratio and detection sensitivity. At the same time, the resonance light scattering detection technology uses white light as the detection light source, the equipment is simple, the price is low, and the detection cost is greatly reduced.
波导（Waveguide）是一种用于微波或可见光波传输的装置，其中使用光纤作为传输媒介的光波导（Optical/light Waveguide）被广泛应用于网络接入和数字通信当中，在不同折色率介质间，由于电磁波的全反射使光波局限在波导及其周围有限区域内传播，当光束由光密介质入射光疏介质时，当且仅当入射角大于临界角时，光信号发生全反射，利用光波导向周围有限区域辐射能量这一原理，将生物芯片作为光波导，激发光由生物芯片边缘大于临界角入射，在生物芯片上下内表面以全反射方式传播，光波导激发方式的优点是激发选择性强，即只激发位于生物芯片表面的发色团，光能量利用率高和系统背景噪声低。Waveguide (Waveguide) is a device for microwave or visible light wave transmission, in which optical waveguide (Optical/light Waveguide) using optical fiber as the transmission medium is widely used in network access and digital communication, in different refractive index media In the meantime, due to the total reflection of the electromagnetic wave, the light wave is confined to the waveguide and its surrounding limited area. When the light beam enters the optically sparse medium from the optically dense medium, the optical signal undergoes total reflection if and only when the incident angle is greater than the critical angle. Using Based on the principle of light waveguide radiating energy to a limited area around it, the biochip is used as an optical waveguide, and the excitation light is incident from the edge of the biochip greater than the critical angle, and propagates in the way of total reflection on the upper and lower inner surfaces of the biochip. The advantage of the optical waveguide excitation method is the excitation selection. Strong performance, that is, only the chromophore located on the surface of the biochip is excited, the utilization rate of light energy is high and the background noise of the system is low.
本发明的有益效果是：本发明利用共振光散射原理，将激发光由生物芯片边缘大于临界角入射，在生物芯片上下内表面以全反射方式传播，通过使用光波导激发、动态扫描和数据拼接，实现了对生物芯片的高效、快速、准确的检测，本发明检测时重现性强并具有很高的灵敏度，有效地降低背景噪声并提高了光信号利用率，同时兼具有低成本的优点。The beneficial effects of the present invention are: the present invention utilizes the principle of resonance light scattering, the excitation light is incident from the edge of the biochip greater than the critical angle, propagates in the way of total reflection on the upper and lower inner surfaces of the biochip, through the use of optical waveguide excitation, dynamic scanning and data splicing , to achieve efficient, fast and accurate detection of biochips, the invention has strong reproducibility and high sensitivity during detection, effectively reduces background noise and improves the utilization rate of optical signals, and at the same time has low cost advantage.
附图说明Description of drawings
图1为本发明的基于共振光散射检测生物芯片的装置的结构示意图：Fig. 1 is the structural representation of the device based on resonance light scattering detection biochip of the present invention:
图2为激发光转换为共振散射光的过程示意图；Figure 2 is a schematic diagram of the process of converting excitation light into resonant scattered light;
图3为上位机软件界面示意图；Fig. 3 is a schematic diagram of the upper computer software interface;
图4为利用本发明的基于共振光散射检测生物芯片的装置对生物芯片进行扫描得到的结果示意图；4 is a schematic diagram of the results obtained by scanning the biochip using the device for detecting the biochip based on resonance light scattering of the present invention;
图5为利用本发明检测和利用现有生物芯片扫描仪检测生物芯片的结果对比示意图，图5a为使用本发明的装置对生物芯片进行扫描得到的图像，图5b为使用商业化扫描仪对生物芯片进行扫描得到的图像。Fig. 5 is a schematic diagram of comparing the results of using the present invention to detect biochips and using existing biochip scanners. Fig. 5a is an image obtained by using the device of the present invention to scan biochips, and Fig. 5b is a biochip using a commercial scanner. The image obtained by scanning the chip.
图中：1、光源驱动器，2、激发光源，3、光纤，4、耦合透镜组，5、生物芯片载物台，6、光电传感器，7、上位机，8、运动平台，9、多轴电机驱动，10、生物芯片，11、样品，12、图像采集，13、扫描控制，14、相机控制，15、导轨控制，16、扫描数据分析，17、数据复位控制。In the figure: 1. Light source driver, 2. Excitation light source, 3. Optical fiber, 4. Coupling lens group, 5. Biochip stage, 6. Photoelectric sensor, 7. Host computer, 8. Motion platform, 9. Multi-axis Motor drive, 10. Biochip, 11. Sample, 12. Image acquisition, 13. Scanning control, 14. Camera control, 15. Rail control, 16. Scanning data analysis, 17. Data reset control.
具体实施方式Detailed ways
以下结合附图对本发明作进一步详细说明。The present invention will be described in further detail below in conjunction with the accompanying drawings.
如图1所示，本发明的基于共振光散射检测生物芯片的装置，主要由光源驱动器1、激发光源2、光纤3、耦合透镜组4、生物芯片载物台5、光电传感器6、上位机7、运动平台8和多轴电机驱动9组成。As shown in Figure 1, the device for detecting biochips based on resonance light scattering of the present invention mainly consists of a light source driver 1, an excitation light source 2, an optical fiber 3, a coupling lens group 4, a biochip stage 5, a photoelectric sensor 6, and a host computer. 7. It consists of a motion platform 8 and a multi-axis motor drive 9.
光源驱动器1用于控制激发光源2产生激发光，使激发光源2的功率连续可调；激发光源2用于产生激发共振光散射的激发光；激发光通过光纤3输出至耦合透镜组4，耦合透镜组4对其进行光束整形，整形后的激发光入射至生物芯片载物台5上；如图2所示，将样品11均匀分布在生物芯片10表面，然后将生物芯片10放置在生物芯片载物台5上，经过光束整形后的激发光以入射角大于临界角的角度从生物芯片10侧边入射，在生物芯片10上下表面进行多次全反射，此时，样品11受到激发光的激发后发生共振光散射，向外辐射特定波长的共振散射光，共振散射光入射至光电传感器6中；光电传感器6对共振散射光进行光电转换，将光信号转换为电信号，将此电信号输出至上位机7中；上位机7中的上位机软件对来自光电传感器6的电信号进行采集、分析并得到样品11的扫描图像。The light source driver 1 is used to control the excitation light source 2 to generate excitation light, so that the power of the excitation light source 2 can be continuously adjusted; the excitation light source 2 is used to generate excitation light that excites resonance light scattering; the excitation light is output to the coupling lens group 4 through the optical fiber 3, coupled The lens group 4 performs beam shaping on it, and the shaped excitation light is incident on the biochip stage 5; as shown in Figure 2, the sample 11 is evenly distributed on the surface of the biochip 10, and then the biochip 10 is placed on the biochip On the stage 5, the excitation light after the beam shaping is incident from the side of the biochip 10 at an angle greater than the critical angle, and undergoes multiple total reflections on the upper and lower surfaces of the biochip 10. At this time, the sample 11 is subjected to the stimulation of the excitation light Resonant light scattering occurs after excitation, and resonant scattered light of a specific wavelength is radiated outward, and the resonant scattered light is incident on the photoelectric sensor 6; the photoelectric sensor 6 performs photoelectric conversion on the resonant scattered light, converts the optical signal into an electrical signal, and converts the electrical signal Output to the host computer 7; the host computer software in the host computer 7 collects and analyzes the electrical signal from the photoelectric sensor 6 and obtains the scanned image of the sample 11.
上位机7通过自身的通用接口与多轴电机驱动9相连，上位机软件控制多轴电机驱动9的运行，多轴电机驱动9与运动平台8通过线缆相连，用于控制运动平台8的移动，运动平台8用于控制生物芯片载物台5的移动，可以根据需要选择不同的运动位移平台，本实施方式中使用三个直线位移平台实现。The host computer 7 is connected to the multi-axis motor drive 9 through its own general interface, and the host computer software controls the operation of the multi-axis motor drive 9, and the multi-axis motor drive 9 is connected to the motion platform 8 through a cable to control the movement of the motion platform 8 , the motion platform 8 is used to control the movement of the biochip stage 5, and different motion displacement platforms can be selected according to the needs. In this embodiment, three linear displacement platforms are used for realization.
如图3所示，为上位机软件的界面，包括：图像采集12、扫描控制13、相机控制14、导轨控制15、扫描数据分析16和数据复位控制17，上位机软件用于显示、分析和保存采集到的信息，并最终形成扫描图像。As shown in Figure 3, it is the interface of the host computer software, including: image acquisition 12, scan control 13, camera control 14, guide rail control 15, scan data analysis 16 and data reset control 17, the host computer software is used for display, analysis and Save the collected information and finally form a scanned image.
本发明的基于共振光散射检测生物芯片的装置的具体检测过程如下：The specific detection process of the device for detecting biochips based on resonance light scattering of the present invention is as follows:
在生物芯片10的本体基片上使用至少一个金属纳米粒子进行标记，将制备好的生物芯片10放置在生物芯片载物台5上，通过上位机软件控制多轴电机驱动9运行并驱动运动平台8移动，运动平台8移动带动生物芯片载物台5移动，对生物芯片10的整个点样区域进行扫描，获得生物芯片10的扫描图像如图4所示，图像中标记的样点清晰、背景成黑色，说明本发明的基于共振光散射检测生物芯片的装置扫描得到的数据背景噪声低，信号强度强。Use at least one metal nanoparticle on the body substrate of the biochip 10 to mark, place the prepared biochip 10 on the biochip stage 5, and control the multi-axis motor drive 9 to run and drive the motion platform 8 through the host computer software Moving, the movement of the motion platform 8 drives the biochip stage 5 to move, and the entire sampling area of the biochip 10 is scanned to obtain a scanned image of the biochip 10 as shown in Figure 4. The marked samples in the image are clear and the background is formed. Black, indicating that the data scanned by the device for detecting biochips based on resonance light scattering of the present invention has low background noise and strong signal intensity.
对上述制备的生物芯片10使用名称为SpotWareTM Colorimetric MicroarrayScanner的生物芯片扫描仪扫描，扫描结果如图5所示，其中，图5a为使用本发明的装置进行扫描得到的图像，图5b为使用生物芯片扫描仪进行扫描得到的图像，从图5可以看出，对相同生物芯片10进行扫描，本发明的装置扫描得到的数据背景噪声更低，而信号强度更强，提高了检测分辨率和灵敏度。The biochip 10 prepared above is scanned by a biochip scanner called SpotWare ™ Colorimetric MicroarrayScanner, and the scanning result is shown in Figure 5, wherein, Figure 5a is an image obtained by scanning using the device of the present invention, and Figure 5b is an image obtained using a biochip scanner. The image scanned by the chip scanner can be seen from Figure 5 that the same biochip 10 is scanned, and the background noise of the data scanned by the device of the present invention is lower, and the signal strength is stronger, which improves the detection resolution and sensitivity .
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