CN112473757B - A microfluidic chip detection system for rapid detection of food safety - Google Patents
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- 238000002347 injection Methods 0.000 claims description 21
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Abstract
本发明公开了一种用于食品安全快速检测的微流控芯片检测系统，涉及微流控芯片领域，包括：上位机、电化学工作站、继电器模块、磁场驱动模块和微流控芯片，上位机连接到继电器模块和电化学工作站，继电器模块连接到磁场驱动模块，磁场驱动模块连接到微流控芯片，微流控芯片连接到电化学工作站；微流控芯片包括上盖板和下基板，上盖板形成有微流通道，微流通道包括多个区域，分离区通过冲洗区连通到检测区，废液区连通到分离区和检测区，检测区中的三电极检测单元连接到电化学工作站，通过磁场驱动模块实现对待测样品的自动化流向控制，同时在不同区域之间设置可编程通断阀门使得各区域相互独立、互不干扰，更方便的对样品进行检测。
The invention discloses a microfluidic chip detection system for rapid detection of food safety, relates to the field of microfluidic chips, and includes: a host computer, an electrochemical workstation, a relay module, a magnetic field drive module and a microfluidic chip, the host computer Connected to the relay module and the electrochemical workstation, the relay module is connected to the magnetic field driving module, the magnetic field driving module is connected to the microfluidic chip, and the microfluidic chip is connected to the electrochemical workstation; the microfluidic chip includes an upper cover plate and a lower substrate, the upper The cover plate is formed with a microfluidic channel, the microfluidic channel includes a plurality of areas, the separation area is connected to the detection area through the flushing area, the waste liquid area is connected to the separation area and the detection area, and the three-electrode detection unit in the detection area is connected to the electrochemical workstation , The automatic flow direction control of the sample to be tested is realized through the magnetic field drive module, and programmable on-off valves are set between different areas to make each area independent and not interfere with each other, and it is more convenient to detect the sample.
Description
技术领域technical field
本发明涉及微流控芯片领域，尤其是一种用于食品安全快速检测的微流控芯片检测系统。The invention relates to the field of microfluidic chips, in particular to a microfluidic chip detection system for rapid detection of food safety.
背景技术Background technique
食品安全是关系到国民生活的大事，同时也是一项国际性难题，造成食品安全问题的因素非常复杂，而食品部门监管时也只能采取抽检的方式进行质量检测。目前，常用食品安全检测方法有气相、液相色谱-质谱分析法和光谱分析法几种，但是这几种方法都存在一定的不足之处，往往需要到专门的实验室中进行操作，这样往往需要很多大型检测设备和大量的检测试剂，检测耗时也比较长，同时需要专业的实验员进行操作，费用较高。Food safety is a major event related to national life, and it is also an international problem. The factors that cause food safety problems are very complex, and the food department can only conduct quality inspections by sampling inspections. At present, there are several common food safety detection methods including gas phase, liquid chromatography-mass spectrometry and spectroscopic analysis, but these methods all have certain shortcomings, and they often need to be operated in specialized laboratories. It requires a lot of large-scale testing equipment and a large number of testing reagents, and the testing takes a long time. At the same time, it requires professional laboratory personnel to operate, and the cost is high.
而微流控技术使快速、准确的检测手段的出现成为可能，微流控技术现已成为一种新颖的传感平台，在该平台上，可以巧妙的集成不同的分析步骤，生物识别材料和合适的传感器，从而产生了新一代传感器，用于检测真菌毒素、食源性病原体、药物和过敏原等，但现有的检测系统自动化程度不高，不能准确控制芯片内待测样品的流向，同时检测效果不理想，与实际偏差较大。Microfluidics has enabled the emergence of fast and accurate detection methods. Microfluidics has now become a novel sensing platform on which different analysis steps, biometric materials and suitable sensors, resulting in a new generation of sensors for the detection of mycotoxins, food-borne pathogens, drugs, and allergens, etc., but the existing detection systems are not highly automated and cannot accurately control the flow of the sample to be tested in the chip. At the same time, the detection effect is not ideal, and there is a large deviation from the actual.
发明内容SUMMARY OF THE INVENTION
本发明人针对上述问题及技术需求，提出了一种用于食品安全快速检测的微流控芯片检测系统，本发明的技术方案如下：In view of the above problems and technical requirements, the present inventor proposes a microfluidic chip detection system for rapid detection of food safety. The technical solution of the present invention is as follows:
一种用于食品安全快速检测的微流控芯片检测系统，包括上位机、电化学工作站、继电器模块、磁场驱动模块和微流控芯片，所述上位机连接到所述继电器模块和所述电化学工作站，所述继电器模块连接到所述磁场驱动模块，所述微流控芯片安装在所述磁场驱动模块中，所述微流控芯片连接到所述电化学工作站；A microfluidic chip detection system for rapid detection of food safety includes a host computer, an electrochemical workstation, a relay module, a magnetic field drive module and a microfluidic chip, the host computer is connected to the relay module and the electrical a chemical workstation, the relay module is connected to the magnetic field driving module, the microfluidic chip is installed in the magnetic field driving module, and the microfluidic chip is connected to the electrochemical workstation;
所述微流控芯片包括上盖板和下基板，所述上盖板和所述下基板键合在一起，所述上盖板形成有微流通道，所述微流通道包括分离区、冲洗区、检测区和废液区，所述分离区通过所述冲洗区连通到所述检测区，所述废液区连通到所述分离区和所述检测区，所述检测区包括两个检测通道和三电极检测单元，所述两个检测通道连通到所述冲洗区，所述三电极检测单元的一端连接到所述两个检测通道中、另一端连接到所述电化学工作站，所述电化学工作站测定所述微流控芯片中待测样品的浓度。The microfluidic chip includes an upper cover plate and a lower substrate, the upper cover plate and the lower substrate are bonded together, the upper cover plate is formed with a microfluidic channel, and the microfluidic channel includes a separation area, a flushing The separation area is connected to the detection area through the flushing area, the waste liquid area is connected to the separation area and the detection area, and the detection area includes two detection areas. a channel and a three-electrode detection unit, the two detection channels are connected to the flushing area, one end of the three-electrode detection unit is connected to the two detection channels, and the other end is connected to the electrochemical workstation, the The electrochemical workstation measures the concentration of the sample to be tested in the microfluidic chip.
其进一步的技术方案为，所述分离区包括微反应池和多个注入口，缓冲溶液注入口、免疫磁珠注入口和待测样品注入口连通到所述微反应池，所述废液区包括第一废液池和第二废液池，所述第一废液池和所述微反应池之间的连接通道上设置有第一阀门，所述冲洗区包括清洗池和清洗液注入口，所述微反应池和所述清洗池之间的连接通道上设置有第二阀门，所述清洗池和所述检测区之间的连接通道上设置有第三阀门，所述冲洗区通过所述两个检测通道连通到所述第二废液池。A further technical solution is that the separation zone includes a micro-reaction cell and a plurality of injection ports, the buffer solution injection port, the immunomagnetic bead injection port and the sample injection port are connected to the micro-reaction cell, and the waste liquid area is connected to the micro-reaction cell. It includes a first waste liquid pool and a second waste liquid pool, a first valve is provided on the connecting channel between the first waste liquid pool and the micro-reaction pool, and the flushing area includes a cleaning pool and a cleaning liquid injection port, A second valve is provided on the connection channel between the micro-reaction pool and the cleaning pool, and a third valve is provided on the connection channel between the cleaning pool and the detection area, and the flushing area passes through the two valves. The detection channels are connected to the second waste liquid pool.
其进一步的技术方案为，所述三电极检测单元包括三个三电极检测阵列，每个三电极检测阵列包括一个参比电极、两个辅助电极和两个工作电极，每个电极都包括连接件和检测件，所述检测件通过所述连接件连接到所述电化学工作站，所述一个参比电极的两个检测件分别位于到所述两个检测通道中，每个检测通道中还设置有一个辅助电极的检测件和一个工作电极的检测件。A further technical solution is that the three-electrode detection unit includes three three-electrode detection arrays, each three-electrode detection array includes a reference electrode, two auxiliary electrodes and two working electrodes, and each electrode includes a connector. and a detection piece, the detection piece is connected to the electrochemical workstation through the connection piece, the two detection pieces of the one reference electrode are respectively located in the two detection channels, and each detection channel is also provided with There is a detection part for the auxiliary electrode and a detection part for the working electrode.
其进一步的技术方案为，参比电极的连接件位于所述两个检测通道之间，第一辅助电极的连接件和第一工作电极的连接件位于所述两个检测通道的一侧，第二辅助电极的连接件和第二工作电极的连接件位于所述两个检测通道的另一侧。A further technical solution is that the connection piece of the reference electrode is located between the two detection channels, the connection piece of the first auxiliary electrode and the connection piece of the first working electrode are located on one side of the two detection channels, and the first auxiliary electrode is located at one side of the two detection channels. The connecting piece of the two auxiliary electrodes and the connecting piece of the second working electrode are located on the other side of the two detection channels.
其进一步的技术方案为，参比电极的检测件的长度大于辅助电极检测件的长度和工作电极检测件的长度之和，辅助电极的检测件横向区域和工作电极的检测件横向区域位于参比电极的检测件横向区域内，且辅助电极的检测件横向区域和工作电极的检测件横向区域不重合。Its further technical scheme is that the length of the detection part of the reference electrode is greater than the sum of the length of the auxiliary electrode detection part and the length of the working electrode detection part, and the lateral area of the detection part of the auxiliary electrode and the lateral area of the detection part of the working electrode are located in the reference electrode. In the lateral area of the detection part of the electrode, and the lateral area of the detection part of the auxiliary electrode and the lateral area of the detection part of the working electrode do not overlap.
其进一步的技术方案为，所述磁场驱动模块包括八个电磁线圈，所述继电器模块包括八个继电器，每个电磁线圈连接到一个继电器，所述上位机控制每个继电器的通断。In a further technical solution, the magnetic field drive module includes eight electromagnetic coils, the relay module includes eight relays, each electromagnetic coil is connected to one relay, and the host computer controls the on-off of each relay.
其进一步的技术方案为，所述测定所述微流控芯片中待测样品的浓度，包括：A further technical solution thereof is that the measuring the concentration of the sample to be tested in the microfluidic chip includes:
制备n个等浓度梯度的标准溶液，各个标准溶液的浓度已知，将各个标准溶液分别经过检测区，控制三电极检测单元的电势以不同的时间速率以三角波形进行扫描，建立不同浓度标准溶液的n个电流-电势曲线；Prepare n standard solutions of equal concentration gradient, the concentration of each standard solution is known, pass each standard solution through the detection area, control the potential of the three-electrode detection unit to scan with a triangular waveform at different time rates, and establish standard solutions of different concentrations n current-potential curves of ;
根据电流-电势曲线获得每个标准溶液的信号强度，通过数据拟合建立不同标准溶液浓度和信号强度大小的对应的标准曲线；Obtain the signal intensity of each standard solution according to the current-potential curve, and establish the corresponding standard curve of different standard solution concentrations and signal intensity by data fitting;
当检测浓度未知的待测样品时，通过所述三电极检测单元检测得到的信号强度，在所述标准曲线中即可获取所述待测样品的浓度。When a sample to be tested with an unknown concentration is detected, the concentration of the sample to be tested can be obtained from the standard curve according to the signal intensity detected by the three-electrode detection unit.
其进一步的技术方案为，所述待测样品浓度的表达式为：Its further technical scheme is that the expression of the concentration of the sample to be tested is:
yt＝a+bxt；y t =a+bx t ;
其中，yt表示待测样品的信号强度，xt表示待测样品的浓度，Among them, y t represents the signal intensity of the sample to be tested, x t represents the concentration of the sample to be tested,
式中，
其中，xi表示第i个标准溶液的浓度，yi表示第i个标准溶液的信号强度。Among them, xi represents the concentration of the ith standard solution, and yi represents the signal intensity of the ith standard solution.
本发明的有益技术效果是：通过磁场驱动模块实现对待测样品的自动化流向控制；同时设置不同区域方便对样品进行检测，达到各个区域互不干扰的目的；通过三电极检测单元能够获取多组数据，检测结果更加准确，操作更加简便。The beneficial technical effects of the present invention are: the automatic flow direction control of the sample to be tested is realized through the magnetic field drive module; at the same time, different areas are set to facilitate the detection of the sample, so as to achieve the purpose of not interfering with each other in each area; multiple sets of data can be acquired through the three-electrode detection unit , the detection results are more accurate and the operation is easier.
附图说明Description of drawings
图1是本申请的检测系统结构示意图。FIG. 1 is a schematic structural diagram of the detection system of the present application.
图2是本申请的微流控芯片的俯视图。FIG. 2 is a top view of the microfluidic chip of the present application.
图3是本申请的微流控芯片的立体图。FIG. 3 is a perspective view of the microfluidic chip of the present application.
图4是本申请的三电极检测单元的立体图。FIG. 4 is a perspective view of the three-electrode detection unit of the present application.
图5是本申请的三电极检测单元的俯视图。FIG. 5 is a top view of the three-electrode detection unit of the present application.
图6是本申请的磁场驱动模块的示意图。FIG. 6 is a schematic diagram of the magnetic field driving module of the present application.
具体实施方式Detailed ways
下面结合附图对本发明的具体实施方式做进一步说明。The specific embodiments of the present invention will be further described below with reference to the accompanying drawings.
如图1所示，一种用于食品安全快速检测微流控芯片检测系统，包括上位机、电化学工作站、继电器模块、磁场驱动模块和微流控芯片，上位机连接到继电器模块和电化学工作站，继电器模块连接到磁场驱动模块，微流控芯片安装在磁场驱动模块中，微流控芯片连接到电化学工作站，上位机用来对整个检测系统进行控制，电化学工作站用来测定微流控芯片中待测样品的浓度。As shown in Figure 1, a microfluidic chip detection system for rapid detection of food safety includes a host computer, an electrochemical workstation, a relay module, a magnetic field drive module and a microfluidic chip. The host computer is connected to the relay module and electrochemical Workstation, the relay module is connected to the magnetic field drive module, the microfluidic chip is installed in the magnetic field drive module, the microfluidic chip is connected to the electrochemical workstation, the host computer is used to control the entire detection system, and the electrochemical workstation is used to measure the microfluidic Control the concentration of the sample to be tested in the chip.
如图2所示，微流控芯片包括上盖板1和下基板2，上盖板由PDMS(聚二甲基硅氧烷)制成，下基板由玻璃材质制成，上盖板1和下基板2键合在一起，上盖板形成有微流通道，微流通道包括分离区3、冲洗区4、检测区5和废液区6，各个区域通过连接通道连通，分离区3通过冲洗区4连通到检测区5，废液区6连通到分离区3和检测区5，检测区5连接到电化学工作站。As shown in Figure 2, the microfluidic chip includes an upper cover 1 and a
优选的，上盖板1和下基板2都为长方体形状，各处连接通道的横截面为长方形，上盖板1的尺寸优选为：长5cm，宽3cm，高4mm；下基板2的尺寸优选为：长5.5cm，宽3.5cm，高1mm；连接通道的横截面尺寸优选为：宽度0.2mm，长度0.4mm。Preferably, both the upper cover plate 1 and the
如图3所示，分离区包括微反应池7和多个注入口，缓冲溶液注入口8、免疫磁珠注入口9和待测样品注入口10连通到微反应池，废液区6包括第一废液池11和第二废液池12，第一废液池11和微反应池7之间的连接通道上设置有第一阀门13，冲洗区4包括清洗池14和清洗液注入口15，微反应池7和清洗池14之间的连接通道上设置有第二阀门16，清洗池14和检测区5的连接通道上设置有第三阀门17，检测区5包括三电极检测单元18、第一检测通道19和第二检测通道20，冲洗区4通过两个检测通道连通到第二废液池12，三电极检测单元18的一端连接到两个检测通道内、另一端连接到电化学工作站。As shown in FIG. 3 , the separation area includes a
当进行检测时，控制第一阀门13开通，第二阀门16关闭，将缓冲溶液从缓冲溶液注入口8注入，这样微反应池7和第一废液池11中将充满缓冲溶液，缓冲溶液起到载体的作用，不参与实际的反应。随后关闭第一阀门13，再将免疫磁珠从免疫磁珠注入口9注入，同时将待测样品从待测样品注入口10注入，注入口的注入工作由多路恒压泵完成注入。When the detection is performed, the
当微反应池7中的反应完成后，此时待测样品附着在免疫磁珠上，此时关闭第三阀门17，打开第一阀门13和第二阀门16，从清洗液注入口15注入清洗液，清洗液会由于缓冲溶液的作用力往第一废液池11流动，此时通过磁场控制模块使免疫磁珠带着待测样品从微反应池7流向清洗池14，达到清洗液清洗的目的，随后关闭第二阀门16，打开第三阀门17，待测样品流向检测区。When the reaction in the
三个阀门都是由金属固体组成的，外形为圆柱体，完全贯穿上盖板，同时能够进行上下移动，圆柱体直径与连接通道宽度相等，三个阀门分别对应三个独立的阀门控制器，根据检测流程，由控制器控制阀门上下移动，实现连接通道内流体的通断，同时为了保证关闭阀门时通道不漏液，需要对其表面进行塑化处理。The three valves are all composed of metal solids. The shape is a cylinder, which completely penetrates the upper cover and can move up and down at the same time. The diameter of the cylinder is equal to the width of the connecting channel. The three valves correspond to three independent valve controllers. According to the detection process, the controller controls the valve to move up and down to realize the on-off of the fluid in the connecting channel. At the same time, in order to ensure that the channel does not leak when the valve is closed, the surface needs to be plasticized.
如图4所示，三电极检测单元包括三个三电极检测阵列，每个三电极检测阵列包括第一参比电极21、第一辅助电极22、第二辅助电极23、第一工作电极24和第二工作电极25，其中每个电极都包括连接件和检测件，检测件通过连接件连接到电化学工作站，一个参比电极的两个检测件分别位于到两个检测通道中，每个检测通道中还设置有一个辅助电极的检测件和一个工作电极的检测件。在微流控芯片的上盖板涂覆有导体材料，由连接件将信号传导上盖板上，随后与电化学工作站连接，其中，参比电极是为了在检测过程中提供一个稳定的电极电势，辅助电极作用是在整个检测过程中与工作电极形成一个回路，工作电极是检测过程中电化学反应发生的区域，由三电极检测阵列可以准确测量电极电势及电势变化。设置三个三电极检测阵列和两个检测通道，实际上相当于获取到六组数据，能够更加精准的获取待测样品的浓度。As shown in FIG. 4 , the three-electrode detection unit includes three three-electrode detection arrays, each of which includes a
如图5所示，第一参比电极的连接件26位于两个检测通道之间，第一辅助电极的连接件27和第一工作电极的连接件28位于两个检测通道的一侧，第二辅助电极的连接件29和第二工作电极的连接件30位于两个检测通道的另一侧，排布更加合理，便于后面与电化学工作站之间的排线布局。As shown in FIG. 5 , the connecting
参比电极的检测件31的长度大于辅助电极检测件32的长度和工作电极检测件33的长度之和，辅助电极的检测件横向区域和工作电极的检测件横向区域位于参比电极的检测件横向区域内，且辅助电极的检测件横向区域和工作电极的检测件横向区域不重合，由此实现每一个三电极检测阵列对特定区域的准确测量，计算更加准确。The length of the
进一步的，如图6所示，磁场驱动模块包括八个电磁线圈34，能够在多个方向上驱动微流控芯片中免疫磁珠的运动，每个电磁线圈连接到一个继电器，因此继电器模块中包括八个继电器，上位机控制继电器模块的通断。Further, as shown in FIG. 6, the magnetic field driving module includes eight
测定浓度的方法包括以下步骤：The method for determining the concentration includes the following steps:
制备n个等浓度梯度的标准溶液，各个标准溶液的浓度已知，将各个标准溶液分别经过检测区，控制三电极检测单元的电势以不同的时间速率以三角波形进行一次或多次反复扫描，电势范围时电极上能交替发生不同的还原反应和氧化反应，建立不同浓度标准溶液的n个电流-电势曲线；Prepare n standard solutions of equal concentration gradient, the concentration of each standard solution is known, pass each standard solution through the detection area respectively, control the potential of the three-electrode detection unit to perform one or more repeated scans with a triangular waveform at different time rates, In the potential range, different reduction reactions and oxidation reactions can alternately occur on the electrode, and n current-potential curves of standard solutions with different concentrations are established;
利用标准溶液相应的信号强度与其浓度成正比关系，根据电流-电势曲线获得每个标准溶液的信号强度，信号强度通常可以是最大电流，即每个标准溶液对应一个最大电流，通过数据拟合建立不同标准溶液浓度和信号强度大小的对应的标准曲线；The corresponding signal intensity of the standard solution is proportional to its concentration, and the signal intensity of each standard solution is obtained according to the current-potential curve. The signal intensity can usually be the maximum current, that is, each standard solution corresponds to a maximum current, which is established by data fitting. Corresponding standard curves of different standard solution concentrations and signal intensities;
当检测浓度未知的待测样品时，通过所述三电极检测单元检测得到的信号强度，在标准曲线中即可获取待测样品的浓度。When a sample to be tested with an unknown concentration is detected, the concentration of the sample to be tested can be obtained from the standard curve by the signal intensity detected by the three-electrode detection unit.
待测样品浓度的表达式为：The expression for the concentration of the sample to be tested is:
yt＝a+bxt；y t =a+bx t ;
其中，yt表示待测样品的信号强度，xt表示待测样品的浓度，Among them, y t represents the signal intensity of the sample to be tested, x t represents the concentration of the sample to be tested,
式中，
其中，xi表示第i个标准溶液的浓度，yi表示第i个标准溶液的信号强度。Among them, xi represents the concentration of the ith standard solution, and yi represents the signal intensity of the ith standard solution.
以上所述的仅是本申请的优选实施方式，本发明不限于以上实施例。可以理解，本领域技术人员在不脱离本发明的精神和构思的前提下直接导出或联想到的其他改进和变化，均应认为包含在本发明的保护范围之内。The above descriptions are only preferred embodiments of the present application, and the present invention is not limited to the above embodiments. It can be understood that other improvements and changes directly derived or thought of by those skilled in the art without departing from the spirit and concept of the present invention should be considered to be included within the protection scope of the present invention.
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