CN110824165B - Lung cancer tumor marker detection device and method based on micro-fluidic chip and mobile phone - Google Patents
Lung cancer tumor marker detection device and method based on micro-fluidic chip and mobile phone Download PDFInfo
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Abstract
The invention discloses a lung cancer tumor marker detection device and a method based on a micro-fluidic chip and a mobile phone.A micro-lens group and a condenser are fixedly connected in a first layer structure of a micro-amplification device, the micro-lens group is arranged under a camera of the smart phone, the condenser is arranged under a flash lamp of the smart phone, a total reflection plane mirror positioned under the micro-fluidic chip is fixedly connected in a second layer structure, a liquid inlet and an observation area are arranged on the upper surface of the micro-fluidic chip, a filter paper, a reaction tank, a capillary pipeline and a waste liquid tank are arranged in the micro-fluidic chip, a layer of filter paper used for stopping blood cells is connected between the reaction tank and the liquid inlet, labeled antibodies of lung cancer markers with different colors of fluorescence are attached to the bottom and the wall of the reaction tank, a detection strip is printed in the middle of the capillary pipeline, and an antibody probe capable of specifically reacting with the lung cancer markers is printed at the bottom of the detection strip; the invention can detect several common lung cancer serum tumor markers and can accurately acquire detection reaction information.
Description
Technical Field
The invention relates to the field of medical detection, aims to detect lung cancer tumor markers, and particularly relates to an early detection method for the lung cancer tumor markers by adopting a smart phone and a micro-fluidic chip, which is beneficial to early screening of lung cancer.
Technical Field
Lung cancer is a killer of cancer, with high morbidity and mortality, and the best way to treat lung cancer is early discovery, early treatment, and thus, earlier discovery is more beneficial for treating lung cancer. An important approach for early cancer detection is tumor screening, and therefore, the method has great significance for popularization of tumor screening and improvement of diagnosis rate.
Traditional tumor screening can only be completed in regular medical institutions by means of large professional instruments and under the operation of professional technicians, such as serum tumor screening, CT, X-ray examination and the like. The document with the Chinese patent application number of 201710011608.8 provides a self-service tumor marker detection device and method, the device can collect fingertip blood, and the fingertip blood and a fluorescent labeled antibody generate an antigen-antibody reaction, and finally color is developed in a detection color band, and then a smart phone is used for directly taking a picture and analyzing to obtain a conclusion. However, since this device simultaneously detects tumor markers of a plurality of cancers, the accuracy of detection cannot be ensured. Secondly, the intelligent mobile phone in the detection mode directly shoots the detection color band, small cell cancers in the tumor marker and the tumor marker with low content in blood are difficult to shoot by the intelligent mobile phone, and the detection result is influenced.
Disclosure of Invention
Aiming at the problems of the existing tumor marker detection, the invention provides a lung cancer tumor marker detection device and method based on a microfluidic chip and a mobile phone.
The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone adopts the technical scheme that: the intelligent mobile phone is horizontally inserted into the upper part of the micro-amplifying device, the intelligent mobile phone is provided with an intelligent mobile phone camera and an intelligent mobile phone flash lamp, a first layer structure above the micro-amplifying device and a second layer structure below the micro-amplifying device are stacked to form the intelligent mobile phone, a micro lens group and a condenser lens are fixedly connected inside the first layer structure, the micro lens group is arranged right below the intelligent mobile phone camera, the condenser lens is arranged right below the intelligent mobile phone flash lamp, and the micro-fluidic chip is horizontally arranged below the micro lens group and the condenser lens; the second layer structure is internally and fixedly connected with a total reflection plane mirror positioned below the microfluidic chip, the total reflection plane mirror is a V-shaped mirror consisting of two plane mirrors forming an angle of 45 degrees with the horizontal plane, and the mirror surfaces of the two plane mirrors are positioned under the micro-lens group and the collecting mirror respectively; the upper surface of the microfluidic chip is provided with a liquid inlet and an observation area, the microfluidic chip is internally provided with filter paper, a reaction tank, a capillary pipeline and a waste liquid tank, a micro-lens group and a smartphone camera are arranged right above the observation area, a layer of filter paper for stopping blood cells is connected between the reaction tank and the liquid inlet, labeled antibodies with lung cancer markers of different colors and fluorescence are attached to the bottom and the wall of the reaction tank, the labeled antibodies can generate antigen-antibody reaction with the lung cancer markers in serum correspondingly, the bottom of the reaction tank is connected with one end of the capillary pipeline, the other end of the capillary pipeline is connected with the waste liquid tank, a detection strip is printed in the middle of the capillary pipeline, an antibody probe capable of specifically reacting with the lung cancer markers is printed at the bottom of the detection strip, and the antibody probe has no cross reaction with the labeled antibodies.
The detection method of the lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone adopts the technical scheme that:
step A: taking blood to be detected to enter from the liquid inlet, filtering the blood by filter paper, reacting serum in the microfluidic chip, and detecting color development in the strip;
and B: open smart mobile phone's camera and smart mobile phone flash light, the detection strip can be clearly seen in manual focusing, takes the photo: step C: processing the photographed detection strip pictures with different colors by adopting app in the smart phone, sequentially calculating corresponding gray values, respectively representing the lung cancer tumor markers by each color strip, and processing each gray valueX i With gray value thresholdN i In comparison, ifX i ≤N i If so, the concentration of the corresponding lung cancer tumor marker is normal, otherwise, the concentration exceeds the standard.
Compared with the traditional detection, the invention has the following advantages:
1. the invention can finish detection only by one smart phone, one microscopic amplification device and one microfluidic chip, can detect several common lung cancer serum tumor markers such as CEA, CA125, NSE, SCC, cyFRA21-1 and the like, can accurately acquire detection reaction information in the microfluidic chip to obtain a more accurate result, has small volume and convenient carrying, realizes the effect of a mobile phone microscope, and overcomes the problem that the traditional detection mode can be operated by professional technicians.
2. The invention comprises a micro lens group packaged by MOEMS, which can automatically correct the image distortion and ensure the clear and stable microscopic imaging. The complicated operation problem that the traditional microscope needs to adjust the aperture, the thick and thin focusing screws is solved.
3. The microfluidic chip provided by the invention contains the reaction pool and the detection strip inside, the antigen-antibody reaction of the lung cancer marker in blood is carried out in the microfluidic chip, and during operation, the blood to be detected is only dripped into the liquid inlet of the chip, and the reaction can be completed without redundant operation. The problem that complex physical and chemical reaction treatment is needed in the traditional detection is solved.
4. According to the invention, a series of detection operations are carried out on the smart phone by adopting the mobile phone app, and the detection operations comprise calling a mobile phone camera to take a picture, carrying out image processing on the taken picture and carrying out comprehensive analysis on the processed data. And a professional instrument is not required for analysis, and the operation is simple. And the detection speed is high, and long-time waiting is not needed. The problems that the traditional detection needs complex processing and long waiting are solved.
5. According to the mobile phone app, the manual focusing algorithm is added, so that the focal length can be adjusted manually, the obtained image is ensured to be clearer, and the problem that the traditional microscopic equipment cannot focus is solved.
6. Due to the popularization of smart phones, the detection device can complete detection only by configuring one microscopic amplification device and one microfluidic chip, so the cost is low. The problem that the traditional detection equipment is expensive is solved.
Drawings
FIG. 1 is a schematic structural diagram of a lung cancer tumor marker detection device based on a microfluidic chip and a mobile phone according to the present invention,
fig. 2 is a side view of the detection apparatus of the smart phone of fig. 1 in a detection state;
FIG. 3 is a perspective view of a first layer structure in the microscopic magnification device of FIG. 1;
FIG. 4 is a perspective view of a second layer structure in the microscopic magnification device of 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 a detection method of the detection device according to the present invention.
The serial numbers and names of the various components in the drawings: 1: microscopic magnification device, 2: microfluidic chip, 3: smart phone, 4: groove, 5: flash lamp hole, 6: camera hole, 7: first layer structure, 8: second layer structure, 9: smart phone camera, 10: smart phone flash light, 11: microlens group, 12: condenser lens, 13: total reflection plane mirror, 14: chip socket, 15: card slot, 16: liquid inlet, 17: filter paper, 18: reaction tank, 19: capillary channel, 20: waste liquid pool, 21: observation zone, 22: detection strip, 23: an antibody probe.
Detailed Description
Referring to fig. 1, fig. 1 shows a lung cancer tumor marker detection device based on a micro-fluidic chip and a mobile phone, which is provided by the invention, and consists of a micro-amplification device 1, a micro-fluidic chip 2 and a smart phone 3. The micro-fluidic chip 2 is horizontally arranged and fixedly inserted in the middle of the micro-amplification device 1, and the smart phone 3 is horizontally inserted in the upper part of the micro-amplification device 1 and positioned above the micro-fluidic chip 2, so that the function of a mobile phone microscope can be realized together with the micro-amplification device 1, and the micro-amplification effect on the micro-fluidic chip 2 is realized.
Referring to fig. 1 and 2, the microscopical device 1 consists of an upper first layer 7 and a lower second layer 8 stacked on top of each other. The first layer 7 and the second layer 8 are all square in shape. The upper part of the first layer structure 7 is provided with a square notch matched with the smart phone 3, so that the smart phone 3 can be horizontally inserted into the upper part of the first layer structure 7. The smart phone 3 is provided with a smart phone camera 9 and a smart phone flash lamp 10, and when the smart phone 3 is inserted into the upper portion of the first layer structure 7, the smart phone camera 9 and the smart phone flash lamp 10 face downwards.
The inside fixed connection micro lens group 11 and condensing lens 12 of first layer structure 7, micro lens group 11 is under smart mobile phone camera 9, and condensing lens 12 is under smart mobile phone flash light 10. The micro-fluidic chip 2 is horizontally arranged below the micro-lens group 11 and the condenser 12, and the micro-fluidic chip 2 is fixedly inserted between the junctions of the first layer structure 7 and the second layer structure 8. The micro-lens group 11 is formed by vertically overlapping five micro-fluidic lenses and is packaged by MOEMS technology, can automatically correct image distortion, can serve as an objective lens of a microscope, and has higher magnification and better magnification effect. The condenser 12 is composed of a convex lens, and can condense the light emitted by the smartphone flash 10, so as to provide a sufficient light source for the microlens assembly 11. The second layer structure 8 is fixedly connected with a total reflection plane mirror 13, and the total reflection plane mirror 13 is positioned below the microfluidic chip 2. The total reflection plane mirror 13 is a V-shaped mirror composed of two plane mirrors forming an angle of 45 degrees with the horizontal plane, and the mirror surfaces of the two plane mirrors are located right below the microlens set 11 and the condenser 12, respectively. When the device works, the camera 9 of the smart phone serves as an ocular lens of a microscope, the micro-lens group 11 serves as an objective lens of the microscope, light emitted by the flash lamp 10 of the smart phone irradiates the total reflection plane mirror 11 through the condensing lens 12, and finally irradiates the micro-fluidic chip 2 through two reflections, so that a sufficient light source is provided for the microscope, and the whole device achieves the effect of a mobile phone microscope.
Referring to fig. 1 and 3, a groove 4 is formed in the upper portion of a first layer structure 7 of the micro-amplification device 1, and the groove 4 is a slightly recessed slot, and mainly functions to place a protruding smartphone camera 9 and a smartphone flash 10, so that the micro-amplification device 1 is better attached to the smartphone 3. The bottom of recess 4 sets up camera hole 6 and flash lamp hole 5, is microlens group 11 under camera hole 6, is condensing lens 12 under flash lamp hole 5. The bottom of the first layer structure 7 is provided with a micro-fluidic chip slot 14, the micro-fluidic chip slot 14 is arranged below the micro-lens group 11 and the condenser 12, and the micro-fluidic chip 2 is inserted into the micro-amplification device 1 from the micro-fluidic chip slot 14.
Referring to fig. 1 and 4, two total reflection plane mirrors 14 and four clamping grooves 15 are fixedly installed on the second layer structure 8 of the microscopic amplification device 1, and the total reflection plane mirrors 14 form an angle of 45 degrees with the horizontal plane, so that light emitted by the smartphone flash 10 which is vertically emitted can be reflected twice and then vertically reflected upwards to the microfluidic chip 2. The clamping groove 15 is used for fixing the micro-fluidic chip 2 to prevent the micro-fluidic chip from sliding and influencing the imaging effect.
Referring to fig. 1 and 5, the microfluidic chip 2 has a liquid inlet 16 and an observation area 21 on the upper surface, and a filter paper 17, a reaction cell 18, a capillary channel 19, and a waste liquid cell 20 inside. Directly above the observation area 21 are the microlens set 11 and the smartphone camera 9, so that the smartphone camera 9 can photograph the observation area 21.
The reaction tank 18 is arranged right below the liquid inlet 16, 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 for stopping blood cells. Blood to be detected enters the microfluidic chip 2 from the liquid inlet 16, is filtered by the filter paper 17, blood cells cannot pass through, and serum enters the reaction tank 18 after being filtered by the filter paper 17. The bottom and wall of the reaction cell 18 are attached with labeled antibodies of lung cancer markers with different colors of fluorescence, and the lung cancer markers in serum such as CEA, CA125, NSE, SCC, cyFRA21-1 and the corresponding labeled antibodies can generate antigen-antibody reaction. The bottom of the reaction tank 18 is connected with one end of a capillary pipeline 19, the other end of the capillary pipeline 19 is connected with a waste liquid tank 20, and a liquid outlet arranged at the bottom of the waste liquid tank 20 is communicated with the outside. A detection strip 22 is printed in the middle of the capillary channel 19, and an antibody probe 23 capable of specifically reacting with a lung cancer marker is pre-printed at the bottom of the detection strip 22.
The serum after the antigen-antibody reaction in the reaction cell 18 flows into the capillary 19, the antibody probe 23 in the capillary 19 does not cross-react with the labeled antibody of the lung cancer marker, and the lung cancer marker reacts with the labeled antibody, so that the detection band 22 is colored. Directly above the detection strip 22 is an observation area 21, and the observation area 21 is directly opposite to the detection strip 22, so that the detection strip 22 can be photographed by the smartphone 3 through the above observation area 21. Different antibody probes 23 are fixed in different areas of the capillary 19 with intervals therebetween, and the antibody probes 23 of a plurality of lung cancer tumor markers such as CEA, CA125, NSE, SCC, cyFRA21-1 form a detection strip 22. The lung cancer marker in the serum passes through the detection strip 22 and undergoes an antigen-antibody reaction with the antibody probe 23, thereby fixing the lung cancer marker in the detection strip 22. Since the labeled antibodies of different lung cancer tumor markers have fluorescence of different colors, the detection strip 22 becomes a detection color band having a color different from each other in each region. Only by observing the position relationship of a certain strip in the detection strip 22, the lung cancer marker detected by the strip can be known. The darker the color of the band, the higher the content of the lung cancer marker. Further, the waste liquid flowing through the capillary 19 flows into the waste liquid pool 20.
Referring to fig. 1-6, when the lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone works, the microscopic amplification device 1 serves as an optical microscope, the smartphone camera 9 serves as an eyepiece, the microlens set 11 serves as an objective lens, and the smartphone flash lamp 10 serves as a light source, so that the function of the optical microscope is achieved.
Blood to be detected is taken and dripped into the liquid inlet 16 of the microfluidic chip 2 by a dropper, after the blood to be detected is filtered by the filter paper 17, serum reacts in the microfluidic chip 2, and after a short waiting, the color is developed in the detection strip 22 of the microfluidic chip 2.
The smart phone 3 and the micro-fluidic chip 2 are respectively inserted into the micro-amplification device 1, and the smart phone camera 9 is just aligned to the observation area 21 on the micro-fluidic chip 2. And opening the app in the mobile phone, wherein the app can call a mobile phone camera, has a manual focusing function, shoots clear and stable images, processes the shot images, and comprehensively analyzes and draws a conclusion.
The camera of the smart phone 3 is turned on, then the smart phone flash 10 is turned on to provide a light source, then manual focusing is used, images are manually focused, the detection strips 22 can be clearly seen, and the images are guaranteed to be clear and stable. Taking a photo, processing the taken pictures of the detection strips 22 with different colors by adopting app, and calculating corresponding gray values of the pictures in sequenceX i Each color bar represents 5 lung cancer tumor markers including CEA, CA125, NSE, SCC and CyFRA21-1, and the gray value thereofX i The size of (b) represents the concentration of each lung cancer tumor marker, and each gray value is automatically obtainedX i With gray value thresholdN i Compared, wherein the gray value threshold valueN i The value of (b) is determined by the concentration threshold of each lung cancer tumor marker and the size of the detection color band area. After comparison, ifX i ≤N i If the concentration of the corresponding lung cancer tumor marker is normal, otherwise, the concentration of the tumor marker exceeds the standard. Passing through i =5 cycles, over-standard lung cancer tumor markers in CEA, CA125, NSE, SCC and CyFRA21-1The detection result can be obtained by recording the substances and then using a logistic regression algorithm to screen the probability of early lung cancer.
Claims (6)
1. The utility model provides a lung cancer tumour marker detection device based on micro-fluidic chip and cell-phone comprises a micro-amplifier device (1), a micro-fluidic chip (2) and smart mobile phone (3), and smart mobile phone (3) level is inserted on the upper portion of micro-amplifier device (1), and smart mobile phone (3) have smart mobile phone camera (9) and smart mobile phone flash light (10), characterized by certainly: the microscopic amplification device (1) is formed by stacking a first layer structure (7) above and a second layer structure (8) below, a micro lens group (11) and a condenser (12) are fixedly connected inside the first layer structure (7), the micro lens group (11) is arranged right below a camera (9) of the smart phone, the condenser (12) is arranged right below a flash lamp (10) of the smart phone, and a microfluidic chip (2) which is horizontally arranged is arranged below the micro lens group (11) and the condenser (12); the second layer structure (8) is internally and fixedly connected with a total reflection plane mirror (13) positioned below the micro-fluidic chip (2), the total reflection plane mirror (13) is a V-shaped mirror consisting of two plane mirrors forming an angle of 45 degrees with the horizontal plane, and the mirror surfaces of the two plane mirrors are positioned under the micro-lens group (11) and the collecting mirror (12) respectively; a liquid inlet (16) and an observation area (21) are formed in the upper surface of the microfluidic chip (2), filter paper (17), a reaction tank (18), a capillary tube (19) and a waste liquid tank (20) are arranged in the micro-fluidic chip, a micro-lens group (11) and a smartphone camera (9) are arranged right above the observation area (21), a layer of filter paper (17) used for stopping blood cells is connected between the reaction tank (18) and the liquid inlet (16), labeled antibodies of lung cancer markers with different colors of fluorescence are attached to the tank bottom and the tank wall in the reaction tank (18), the labeled antibodies correspondingly react with the lung cancer markers in blood in an antigen-antibody reaction mode, one end of the capillary tube (19) is connected to the bottom of the reaction tank (18), the other end of the capillary tube (19) is connected to the waste liquid tank (20), a detection strip (22) is printed in the middle of the capillary tube (19), an antibody probe (23) capable of specifically reacting with the lung cancer markers is printed at the bottom of the detection strip (22), and the antibody probe (23) has no cross reaction with the labeled antibodies.
2. The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone according to claim 1, wherein: the detection strip (22) is formed by a plurality of antibody probes (23), and the different antibody probes (23) are spaced apart from each other.
3. The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone of claim 1, wherein: the micro-fluidic chip (2) is fixedly inserted at the junction of the first layer structure (7) and the second layer structure (8).
4. The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone according to claim 1, wherein: the micro-lens group (11) is formed by vertically stacking five micro-fluidic lenses, and the five micro-fluidic lenses are packaged by MOEMS technology.
5. The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone according to claim 1, wherein: the micro-fluidic chip microscope is characterized in that a groove (4) is formed in the upper portion of the first layer structure (7), a smart phone camera (9) and a smart phone flash lamp (10) are placed in the groove (4), a micro-fluidic chip slot (14) is formed in the bottom of the first layer structure (7), the micro-fluidic chip slot (14) is formed below the micro-lens group (11) and the condenser lens (12), and the micro-fluidic chip (2) can be inserted into the micro-amplification device (1) from the micro-fluidic chip slot (14).
6. The lung cancer tumor marker detection device based on the microfluidic chip and the mobile phone of claim 5, wherein: the bottom of recess (4) sets up camera hole (6) and flash light hole (5), is microlens group (11) under camera hole (6), is condensing lens (12) under flash light hole (5).
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| CN111693694B (en) * | 2020-05-19 | 2023-02-17 | 江苏大学 | Multi-virus immunodetection device and method based on smart phone and micro-fluidic chip |
| CN112240884A (en) * | 2020-09-30 | 2021-01-19 | 兰州大学 | Enzyme activity detection device and detection method based on smart phone and microchip |
| CN114216901B (en) * | 2021-11-22 | 2024-03-19 | 江苏大学 | Sperm activity detection system and method based on microfluidic chip composite smart phone |
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