CN110824165B - Lung cancer tumor marker detection device and method based on microfluidic chip and mobile phone - Google Patents

Abstract

The invention discloses a lung cancer tumor marker detection device and method based on a microfluidic chip and a mobile phone. The first layer structure of the microscopic magnification device is fixedly connected to a microlens group and a condenser lens, and the microlens group is directly below the smartphone camera. , the condenser is directly under the flashlight of the smartphone, and the second layer of structure is fixedly connected to the total reflection plane mirror located under the microfluidic chip. The upper surface of the microfluidic chip is provided with a liquid inlet and an observation area, and there are filter paper, Reaction tank, capillary and waste liquid tank, a layer of filter paper used to stop blood cells is connected between the reaction tank and the liquid inlet, and labeled antibodies of lung cancer markers with different colors of fluorescence are attached to the bottom and wall of the reaction tank , a detection strip is printed in the middle of the capillary, and the bottom of the detection strip is printed with an antibody probe that can specifically react with lung cancer markers; the present invention can detect several common lung cancer serum tumor markers, and can detect Accurate acquisition of reaction information.

Description

Lung cancer tumor marker detection device and method based on microfluidic chip and mobile phone

technical field

The invention relates to the field of medical detection, and is aimed at the detection of lung cancer tumor markers, and specifically uses a smart phone and a microfluidic chip for early detection of lung cancer tumor markers, which is helpful for early screening of lung cancer.

technical background

Lung cancer is a cancer killer with high morbidity and mortality. The best way to treat lung cancer is early detection and early treatment. Therefore, the earlier the detection, the more beneficial it is for the treatment of lung cancer. An important way to detect cancer early is tumor screening. Therefore, it is of great significance to popularize tumor screening and improve the diagnosis rate.

Traditional tumor screening can generally only be done in formal medical institutions with the help of large-scale professional equipment and under the operation of professional technicians, such as serum tumor screening, CT, X-ray examination, etc. Chinese patent application No. 201710011608.8 proposes a self-service tumor marker detection device and method. The device can collect fingertip blood, undergo antigen-antibody reaction with fluorescently labeled antibodies, and finally detect tumor markers in the detection ribbon. Color development, and then use the smartphone to directly take pictures, analyze and draw conclusions. However, since the device simultaneously detects tumor markers of multiple cancers, the accuracy of the detection cannot be guaranteed. Secondly, in this detection method, the smartphone directly takes pictures of the detection ribbon, and the small cell cancer and tumor markers with low content in the blood are difficult to be photographed by the smartphone, which affects the detection results.

Contents of the invention

Aiming at the above-mentioned problems existing in the detection of existing tumor markers, the present invention proposes a lung cancer tumor marker detection device and method based on a microfluidic chip and a mobile phone. By detecting multiple lung cancer tumor markers, and analyzing the detection results Comprehensive analysis can draw a more accurate result.

The technical scheme adopted by the lung cancer tumor marker detection device based on the microfluidic chip and mobile phone in the present invention is: it consists of a microscopic amplification device, a microfluidic chip and a smart phone, and the smart phone is horizontally inserted into the microscopic On the upper part of the magnifying device, the smart phone comes with a smart phone camera and a smart phone flash light. The first layer structure above the micro magnifying device and the second layer structure below are stacked. The first layer structure is fixedly connected to the micro lens group and The condenser, the microlens group is directly below the camera of the smartphone, the condenser is directly below the flashlight of the smartphone, and the microfluidic chip is placed horizontally under the microlens group and the condenser; the internal fixed connection of the second layer structure is located in the microfluidic chip The total reflection plane mirror below, the total reflection plane mirror is a V-shaped mirror composed of two plane mirrors at an angle of 45° to the horizontal plane. There is a liquid inlet and an observation area on the surface, and a filter paper, a reaction pool, a capillary pipe and a waste liquid pool are arranged inside, and a microlens group and a smart phone camera are directly above the observation area, and a connection between the reaction pool and the liquid inlet A layer of filter paper is used to stop blood cells. Labeled antibodies of lung cancer markers with different colors of fluorescence are attached to the bottom and wall of the reaction tank. The labeled antibodies will react with the lung cancer markers in the serum correspondingly. Antigen-antibody reaction, reaction One end of the capillary is connected to the bottom of the pool, and the other end of the capillary is connected to the waste liquid pool. A detection strip is printed in the middle of the capillary, and an antibody probe that can specifically react with lung cancer markers is printed on the bottom of the detection strip. The antibody probe and The labeled antibody has no cross reaction.

The technical scheme adopted by the detection method of the lung cancer tumor marker detection device based on the microfluidic chip and mobile phone is:

Step A: Take the blood to be tested and enter it through the liquid inlet, and after filtering through filter paper, the serum reacts in the microfluidic chip, and the color is developed in the detection strip;

Step B: Turn on the camera of the smartphone and the flashlight of the smartphone, manually adjust the focus, and the detection bands can be clearly seen, and take a photo: Step C: Use the app in the smartphone to compare the pictures of the detection bands of different colors Processing, calculate its corresponding gray value in turn, each color band represents each lung cancer tumor marker, compare each gray value X _i with the gray value threshold N _i , if X _i ≤ N _i , it means The concentration of corresponding lung cancer tumor markers is normal, otherwise it exceeds the standard.

Compared with traditional detection, the present invention has the following advantages:

1. The present invention only needs a smart phone, a microscopic amplification device, and a microfluidic chip to complete the detection, and can target several common lung cancer serum tumor markers such as CEA, CA125, NSE, SCC, and CyFRA21-1 The detection can accurately obtain the detection reaction information in the microfluidic chip, and obtain more accurate results. It is small in size, easy to carry, and realizes the effect of "mobile phone microscope". operational problems.

2. The present invention includes a MOEMS packaged microlens group, which can automatically correct image distortion and ensure clear and stable microscopic imaging. It breaks through the complex operation problems of traditional microscopes that need to adjust the aperture, coarse and fine focus spirals.

3. The microfluidic chip of the present invention contains a reaction pool and a detection strip inside. The antigen-antibody reaction of the lung cancer markers in the blood is placed in the microfluidic chip for operation. Just drop the blood into the liquid inlet of the chip, and the reaction can be completed without redundant operations. It breaks through the problem that traditional detection needs to do complex physical and chemical reaction processing.

4. The present invention uses a mobile phone app to perform a series of detection operations on the smart phone, including calling the camera of the mobile phone to take pictures, performing image processing on the captured pictures, and comprehensively analyzing the processed data. There is no need to use professional instruments for analysis, and the operation is simple. And the detection speed is fast, and there is no need to wait for a long time. It breaks through the problem that traditional detection requires complex processing and long waiting.

5. A manual focusing algorithm is added to the mobile app of the present invention, which can artificially adjust the focal length to ensure that the obtained image is clearer, breaking through the problem that traditional microscopic equipment cannot focus.

6. Due to the popularization of smart phones, the detection device of the present invention only needs to be equipped with a microscopic amplification device and a microfluidic chip to complete the detection, so the cost is relatively low. It breaks through the expensive problem of traditional testing equipment.

Description of drawings

Fig. 1 is the structure schematic diagram of the lung cancer tumor marker detection device based on microfluidic chip and mobile phone according to the present invention,

Fig. 2 is a side view of the detection device of the smartphone in the detection state in Fig. 1;

Fig. 3 is a perspective view of the first layer structure in the micromagnification device in Fig. 1;

Fig. 4 is a perspective view of the second layer structure in the micromagnification device in Fig. 1;

FIG. 5 is an enlarged schematic view of the three-dimensional structure of the microfluidic chip in FIG. 1;

Fig. 6 is a flow chart of the detection method of the detection device of the present invention.

The serial numbers and names of the components in the attached drawings: 1: Microscopic magnification device, 2: Microfluidic chip, 3: Smartphone, 4: Groove, 5: Flash hole, 6: Camera hole, 7: First layer structure , 8: second layer structure, 9: smartphone camera, 10: smartphone flash, 11: microlens group, 12: condenser mirror, 13: total reflection plane mirror, 14: chip slot, 15: card slot, 16: input Liquid port, 17: filter paper, 18: reaction pool, 19: capillary channel, 20: waste liquid pool, 21: observation area, 22: detection band, 23: antibody probe.

detailed description

Referring to FIG. 1 , FIG. 1 is a lung cancer tumor marker detection device based on a microfluidic chip and a mobile phone according to the present invention, which consists of a microscopic amplification device 1 , a microfluidic chip 2 and a smart phone 3 . The microfluidic chip 2 is set horizontally, fixedly inserted in the middle of the micromagnifying device 1, and the smart phone 3 is horizontally inserted in the upper part of the micromagnifying device 1, located above the microfluidic chip 2, and can be realized together with the micromagnifying device 1. The function of the mobile phone microscope realizes the microscopic magnification effect on the microfluidic chip 2 .

Referring to FIG. 1 and FIG. 2 , the micromagnification device 1 is composed of a first layer structure 7 above and a second layer structure 8 below. The outer shapes of the first layer structure 7 and the second layer structure 8 are all square shapes. A square notch matching the smart phone 3 is formed on the top of the first layer structure 7 so that the smart phone 3 can be horizontally inserted into the top of the first layer structure 7 . The smart phone 3 is equipped with a smart phone camera 9 and a smart phone flash 10, and when inserted in the upper part of the first layer structure 7, the smart phone camera 9 and the smart phone flash 10 are directed downward.

The inside of the first layer structure 7 is fixedly connected with the microlens group 11 and the condenser lens 12 , the microlens group 11 is directly below the camera 9 of the smartphone, and the condenser lens 12 is directly below the flashlight 10 of the smartphone. Below the microlens group 11 and the condenser lens 12 is a horizontally placed microfluidic chip 2 , and the microfluidic chip 2 is fixedly inserted between the junction of the first layer structure 7 and the second layer structure 8 . The microlens group 11 is composed of five microfluidic lenses stacked up and down, and is packaged by MOEMS technology, which can automatically correct image distortion. It is an objective lens that can act as a "microscope", with higher magnification and better magnification effect . The condenser lens 12 is composed of a convex lens, which can gather the light emitted by the flashlight 10 of the smart phone and provide sufficient light source for the microlens group 11 . The second layer structure 8 is fixedly connected with a total reflection plane mirror 13 , and the total reflection plane mirror 13 is located below the microfluidic chip 2 . The total reflection plane mirror 13 is a V-shaped mirror composed of two plane mirrors at an angle of 45° to the horizontal plane. The mirror surfaces of the two plane mirrors are located directly below the microlens group 11 and the condenser lens 12 respectively. When working, the smart phone camera 9 acts as the eyepiece of the "microscope", and the microlens group 11 is the "objective lens" of the microscope. The light emitted by the smart phone flash 10 is irradiated on the total reflection plane mirror 11 through the condenser lens 12, and after two reflections, it finally shines on the On the microfluidic chip 2, sufficient light source is provided for the "microscope", so the whole device realizes the function of a mobile phone microscope.

Referring to Fig. 1 and Fig. 3, the upper part of the first layer structure 7 of the micromagnifying device 1 has a groove 4, and the groove 4 is a slightly concave slot, the main function of which is to place the protruding smart phone camera 9 and the smart phone flash 10, Make the microscopic magnifying device 1 fit the smart phone 3 better. A camera hole 6 and a flashlight hole 5 are arranged at the bottom of the groove 4 , a microlens group 11 is directly below the camera hole 6 , and a condenser lens 12 is directly below the flashlight hole 5 . The bottom of the first layer structure 7 is provided with a microfluidic chip slot 14, the microfluidic chip slot 14 is below the microlens group 11 and the condenser lens 12, and the microfluidic chip 2 is inserted into the display from the microfluidic chip slot 14. Micro amplification device 1.

Referring to Fig. 1 and Fig. 4, two total reflection flat mirrors 14 and four draw-in slots 15 are fixedly installed on the second layer structure 8 of the micromagnification device 1, and the total reflection flat mirror 14 forms an angle of 45° with the horizontal plane, and can vertically shoot down The light emitted by the flashlight 10 of the smart phone is reflected twice, and then vertically upwards to the microfluidic chip 2 . The function of the slot 15 is to fix the microfluidic chip 2 to prevent it from sliding and affect the imaging effect.

1 and 5, the upper surface of the microfluidic chip 2 has a liquid inlet 16 and an observation area 21, and a filter paper 17, a reaction pool 18, a capillary channel 19 and a waste liquid pool 20 are arranged inside. Directly above the observation area 21 is a microlens group 11 and a smart phone camera 9, so that the smart phone camera 9 can take pictures of the observation area 21.

The reaction tank 18 is directly below the liquid inlet 16, and a layer of filter paper 17 is connected between the reaction tank 18 and the liquid inlet 16, and the filter paper 17 is used to stop blood cells. The blood to be tested enters the microfluidic chip 2 from the liquid inlet 16 and is filtered through the filter paper 17 , blood cells cannot pass through, and the serum enters the reaction pool 18 after being filtered through the filter paper 17 . A variety of labeled antibodies of lung cancer markers with different colors of fluorescence are attached to the bottom and wall of the reaction tank 18, and the lung cancer markers in the serum such as CEA, CA125, NSE, SCC, CyFRA21-1 and the corresponding labeled antibodies will be An antigen-antibody reaction occurs. One end of the capillary pipe 19 is connected to the bottom of the reaction pool 18, and the other end of the capillary pipe 19 is connected to the waste liquid pool 20, and the bottom of the waste liquid pool 20 is provided with a liquid outlet to communicate with the outside. A detection strip 22 is printed in the middle of the capillary 19, and the bottom of the detection strip 22 is pre-printed with an antibody probe 23 capable of specifically reacting with lung cancer markers.

In the reaction pool 18, the serum after the antigen-antibody reaction flows into the capillary 19, and the antibody probe 23 in the capillary 19 has no cross-reaction with the labeled antibody of the above-mentioned lung cancer marker, and the lung cancer marker has not cross-reacted with the labeled antibody. The reaction thus causes the detection band 22 to develop color. Immediately above the detection strip 22 is an observation area 21, and the observation area 21 is facing the detection strip 22. Therefore, the detection strip 22 can be photographed with the smart phone 3 through the observation area 21 above. Different antibody probes 23 are immobilized in different regions of the capillary 19 with intervals between them, and the antibody probes 23 of various lung cancer tumor markers such as CEA, CA125, NSE, SCC, and CyFRA21-1 form detection bands 22 . The lung cancer markers in the serum will react with the antibody probe 23 after passing through the detection strip 22 , thereby immobilizing the lung cancer markers in the detection strip 22 . Since the labeled antibodies of different lung cancer tumor markers have different colors of fluorescence, the detection strip 22 will become a detection strip with different colors for each region. Only by observing the positional relationship of a certain band in the detection band 22, the lung cancer marker detected by the band can be known. The darker the color of the band, the higher the content of the lung cancer marker. Furthermore, the waste liquid flowing through the capillary channel 19 flows into the waste liquid pool 20 .

Referring to Figures 1-6, when the lung cancer tumor marker detection device based on the microfluidic chip and mobile phone according to the present invention is working, the micromagnifying device 1 acts as an optical microscope, and the smart phone camera 9 is used as the eyepiece. The microlens group 11 is used as the objective lens, and the smart phone flashlight 10 is used as the light source to realize the function of an optical microscope.

Take the blood to be tested and drop it into the liquid inlet 16 of the microfluidic chip 2 with a dropper. After the blood to be tested is filtered by the filter paper 17, the serum reacts in the microfluidic chip 2. Color develops in the detection strip 22 of the chip 2 .

The smartphone 3 and the microfluidic chip 2 are respectively inserted into the micromagnification device 1 , and the smartphone camera 9 is just aligned with the observation area 21 on the microfluidic chip 2 . Open the app in the mobile phone, the app can call the mobile phone camera, and has the function of manual focus, take clear and stable images, perform image processing on the captured pictures, comprehensively analyze and draw conclusions.

Turn on the camera of the smart phone 3, then turn on the flash 10 of the smart phone to provide a light source, and then use manual focus to manually focus the image, and the detection band 22 can be clearly seen to ensure a clear and stable image. Take a picture, use the app to process the pictures of the detection strips 22 of different colors, and calculate the corresponding gray value X _i in turn, and each color strip represents CEA, CA125, NSE, SCC, CyFRA21-1, a total of 5 Lung cancer tumor markers, the size of the gray value X _i represents the concentration of each lung cancer tumor marker, and each gray value X _i is automatically compared with the gray value threshold N _i , where the gray value threshold N _i The value of is determined by the concentration threshold of each lung cancer tumor marker and the size of the detection color band. After comparison, if Xi ≤ N _{i ,} it means that the concentration of the corresponding lung cancer tumor marker is normal; otherwise, the concentration of the tumor marker exceeds the standard. Through i=5 cycles, write down the lung cancer tumor markers that exceed the standard in CEA, CA125, NSE, SCC, and CyFRA21-1, and then use the logistic regression algorithm on them to obtain the detection results and screen for early lung cancer. probability.

Claims (6)

1. A lung cancer tumor marker detection device based on a microfluidic chip and a mobile phone, consisting of a microscopic amplification device (1), a microfluidic chip (2) and a smartphone (3), the smartphone (3) Inserted horizontally on the upper part of the microscopic magnification device (1), the smart phone (3) has its own smart phone camera (9) and smart phone flashlight (10), which is characterized in that the microscopic magnification device (1) consists of the upper One-layer structure (7) and the lower second-layer structure (8) are stacked. The first-layer structure (7) is fixedly connected to the micro-lens group (11) and the condenser lens (12). The micro-lens group (11) is in the smart Directly below the mobile phone camera (9), the condenser lens (12) is directly below the smart phone flashlight (10), and below the microlens group (11) and the condenser lens (12) is a horizontally placed microfluidic chip (2); The two-layer structure (8) is fixedly connected to the total reflection plane mirror (13) located below the microfluidic chip (2). The total reflection plane mirror (13) is a V-shaped mirror composed of two plane mirrors at an angle of 45° to the horizontal plane. The mirror surfaces of the two plane mirrors are located directly below the microlens group (11) and the condenser lens (12); the upper surface of the microfluidic chip (2) is provided with a liquid inlet (16) and an observation area (21). There are filter paper (17), reaction pool (18), capillary pipe (19) and waste liquid pool (20), and the microlens group (11) and smart phone camera (9) are directly above the observation area (21). A layer of filter paper (17) used to stop blood cells is connected between the pool (18) and the liquid inlet (16), and the markers of lung cancer markers with fluorescent colors of different colors are attached to the bottom and wall of the reaction pool (18) Antibody, the labeled antibody will react with the lung cancer markers in the serum correspondingly. The bottom of the reaction pool (18) is connected to one end of the capillary (19), and the other end of the capillary (19) is connected to the waste liquid pool (20). The capillary A detection strip (22) is printed in the middle of the lane (19), and an antibody probe (23) capable of specifically reacting with lung cancer markers is printed on the bottom of the detection strip (22), and the antibody probe (23) is compatible with the The labeled antibody has no cross-reactivity. 2. The lung cancer tumor marker detection device based on microfluidic chip and mobile phone according to claim 1, characterized in that: the detection strip (22) is formed by a variety of antibody probes (23), and different antibody probes (23) with intervals in between. 3. The lung cancer tumor marker detection device based on a microfluidic chip and a mobile phone according to claim 1, characterized in that: the microfluidic chip (2) is fixedly inserted into the first layer structure (7) and the second layer structure ( 8) Junction. 4. The lung cancer tumor marker detection device based on microfluidic chip and mobile phone according to claim 1, characterized in that: the microlens group (11) is composed of five microfluidic lenses stacked up and down, five microfluidic lenses The lens is encapsulated by MOEMS technology. 5. The lung cancer tumor marker detection device based on microfluidic chip and mobile phone according to claim 1, characterized in that: the upper part of the first layer structure (7) has a groove (4), the groove ( 4) A smartphone camera (9) and a smartphone flash (10) are placed in the center, and a microfluidic chip slot (14) is opened at the bottom of the first layer structure (7), and the microfluidic chip slot (14) is in the microlens Below the group (11) and the condenser lens (12), the microfluidic chip (2) can be inserted into the micromagnification device (1) from the microfluidic chip slot (14). 6. The lung cancer tumor marker detection device based on microfluidic chip and mobile phone according to claim 5, characterized in that: the bottom of the groove (4) is provided with a camera hole (6) and a flashlight hole (5), and the camera hole Just below (6) is the microlens group (11), and just below the flash hole (5) is the condenser lens (12).

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