A nuclear pore membrane biochip multi-link detection device and use method

Abstract

The present invention discloses a nuclear pore membrane biochip multiple detection device, including a collection device and a detection device, the collection device includes a first nuclear pore membrane, the first surface of the first nuclear pore membrane faces the body fluid flowing through the first nuclear pore membrane, so as to enrich the cells used for detection in the body fluid on the first surface; the detection device includes a preservation liquid cup containing a preservation liquid, the first surface of the first nuclear pore membrane can be immersed in the preservation liquid cup, and the preservation liquid is used to protect the cell morphology; the second nuclear pore membrane is detachably arranged at the bottom of the preservation liquid cup, and is used to filter the preservation liquid to enrich the cells in the preservation liquid on the first side of the second nuclear pore membrane. The present invention uses the first nuclear pore membrane to enrich the cells to be detected in the body fluid, and transfers them to the preservation liquid for exfoliated cell morphology detection, and uses the second nuclear pore membrane to enrich the cells in the preservation liquid for immunodetection or FISH detection, so as to realize multiple detection in combination with the exfoliated cell morphology detection, and more accurately detect the lesions in the body.

Description

Technical Field

[0001] The present invention relates to the field of human pathology detection equipment, and in particular to a nuclear pore membrane biochip multi-link detection device and a use method thereof. Background Art

[0002] For the detection of various lesions inside the human body, imaging examination combined with intracavitary instrument examination and living tissue pathological diagnosis can accurately determine whether lesions have occurred in the patient's body. However, the above-mentioned examination methods will not only cause physical pain to the patients, but are also not suitable for screening and surveys of tumor diseases.

[0003] In order to more conveniently determine whether a lesion has occurred in the patient's body, the prior art often determines the lesion by means of body fluid detection. One method is to perform a separate exfoliated cell morphological test on the cells in the body fluid. This detection method can conveniently and quickly detect lesions without causing pain to the patient, and has the characteristics of high specificity. However, due to the small amount of exfoliated cells obtained by the centrifugation method due to the limited volume of the centrifuged liquid or the high differentiation of the cells, it will cause low sensitivity and false negative problems; another method is to perform immune detection and FISH (fluorescence in situ hybridization) molecular detection on the cells in the body fluid. This detection method has high sensitivity, but low specificity and cumbersome operation. In the process of making the test slide, it is necessary to obtain the exfoliated cells and smear the cells on the adhesive slide, and go through several cumbersome steps, and each step of the processing needs to be cleaned with a cleaning solution, which may cause cell loss, and because the centrifugation method is used to obtain the exfoliated cells, it will cause cell deformation and affect the results of immune detection and FISH molecular detection.

[0004] Therefore, how to reduce the operational difficulty of lesion detection and improve the accuracy of lesion detection is a technical problem that technical personnel in this field urgently need to solve. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a nuclear pore membrane biochip multi-detection device to reduce the operational difficulty of lesion detection and improve the accuracy of lesion detection.

[0006] Another object of the present invention is to provide a method for using the above-mentioned nuclear pore membrane biochip multiplex detection device to perform multiplex detection.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] A nuclear pore membrane biochip multi-link detection device, comprising a collection device and a detection device, wherein the collection device comprises a first nuclear pore membrane, wherein a first surface of the first nuclear pore membrane faces a body fluid flowing through the first nuclear pore membrane, so as to enrich cells for detection in the body fluid on the first surface;

[0009] The detection device comprises:

[0010] a preservation liquid cup containing a preservation liquid, wherein the first surface can be immersed in the preservation liquid cup, and the preservation liquid is used to protect the cell morphology;

[0011] The second nuclear pore membrane is detachably disposed at the bottom of the preservation solution cup, and the second nuclear pore membrane is used to filter the preservation solution to enrich the cells used for detection in the preservation solution on the first side of the second nuclear pore membrane.

[0012] Preferably, in the above-mentioned nuclear pore membrane biochip multi-detection device, the first side of the second nuclear pore membrane is divided into at least two micropore areas by a cofferdam to separate the cells enriched on the first side of the second nuclear pore membrane into at least two areas for performing different types of detection.

[0013] Preferably, in the above-mentioned nuclear pore membrane biochip multi-detection device, the second nuclear pore membrane is detachably arranged at the bottom of the preservation liquid cup through the reaction pool, the reaction pool is detachably installed at the bottom opening of the preservation liquid cup, and the bottom of the reaction pool can be opened to discharge the preservation liquid in the preservation liquid cup, and the second nuclear pore membrane is fixedly connected to the top of the reaction pool.

[0014] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the first nuclear pore membrane is fixedly connected to the first side of the lower cylinder to enrich the cells for detection in the body fluid flowing through the lower cylinder.

[0015] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, a truncated cone-shaped flow channel is provided inside the lower cylinder, so that the body fluid flowing through the lower cylinder flows out of the lower cylinder from the smaller side of the cone.

[0016] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the outer surface of the first side of the lower cylinder is provided with an external thread, and the top of the preservation liquid cup is provided with an internal thread for threaded connection with the lower cylinder.

[0017] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the collection device further comprises an upper cylinder, one side of which is provided with an internal thread that cooperates with the first side thread of the lower cylinder.

[0018] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the dam is a high molecular polymer layer printed and deposited on the first side of the second nuclear pore membrane.

[0019] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the reaction pool is threadedly connected to the bottom opening of the preservation liquid cup.

[0020] Preferably, in the above-mentioned nuclear pore membrane biochip multiplex detection device, the first nuclear pore membrane is fixed to one side of the lower cylinder by hot-melt welding or adhesive.

[0021] The present invention also provides a method for using a nuclear pore membrane biochip multi-link detection device, wherein the method uses a nuclear pore membrane biochip multi-link detection device having one or more of the above-mentioned technical effects to perform multi-link detection, and the method at least comprises the following steps:

[0022] Enriching body fluid cells: using the collection device to enrich cells in the body fluid on the first surface of the first nuclear pore membrane;

[0023] Transferring cells: turning over the collection device so that the first surface of the first nuclear pore membrane faces the opening of the preservation liquid cup, and immersing the first surface of the first nuclear pore membrane into the preservation liquid in the preservation liquid cup, and transferring the cells on the first surface to the preservation liquid in the preservation liquid cup for morphological detection of exfoliated cells;

[0024] Enriching the preservation solution cells: The cells remaining in the preservation solution after the cell morphology detection are enriched to the first side of the second nuclear pore membrane through the filtering effect of the second nuclear pore membrane, so as to be used for immunodetection or FISH molecular detection.

[0025] Preferably, in the method for using the above-mentioned nuclear pore membrane biochip multiplex detection device, the cell transfer step specifically includes:

[0026] Turning over the collection device so that the first surface of the first nuclear pore membrane faces the opening of the preservation liquid cup, and immersing the first surface of the first nuclear pore membrane in the preservation liquid in the preservation liquid cup;

[0027] The preservation solution cup is shaken using a shaker to transfer the cells on the first surface of the first nucleopore membrane into the preservation solution.

[0028] It can be seen from the above technical scheme that the nuclear pore membrane biochip multi-detection device provided by the present invention includes a collection device and a detection device, wherein the collection device includes a first nuclear pore membrane, the first surface of the first nuclear pore membrane faces the body fluid flowing through the first nuclear pore membrane, so as to enrich the cells for detection in the body fluid on the first surface; the detection device includes a preservation liquid cup containing preservation liquid, the opening of the preservation liquid cup is larger than the area of ​​the first nuclear pore membrane, so that the first surface of the first nuclear pore membrane can be immersed in the preservation liquid cup, and the preservation liquid in the preservation liquid cup is used to protect the cell morphology; the second nuclear pore membrane, the second nuclear pore membrane is detachably arranged at the bottom of the preservation liquid cup, and is used to filter the preservation liquid to enrich the cells for detection in the preservation liquid on the first side of the second nuclear pore membrane. The nuclear pore membrane biochip multi-link detection device provided by the present invention enriches the cells for detection in the body fluid on the first surface of the first nuclear pore membrane through the filtering effect of the first nuclear pore membrane. After the enrichment is completed, the first surface of the first nuclear pore membrane is immersed in a preservation liquid cup containing a preservation liquid to transfer the cells on the first surface to the preservation liquid in the detection device. Compared with the prior art that uses centrifugation to enrich cells in body fluid, the nuclear pore membrane biochip multi-link detection device provided by the present invention uses the first nuclear pore membrane for enrichment and transfer, and the number of cells with intact morphology is larger. The preservation liquid is absorbed for morphological detection of exfoliated cells, and the number of intact cells is larger, thereby improving the detection sensitivity and the detection accuracy; the detection device is detachable A second nuclear pore membrane is arranged on the first side of the second nuclear pore membrane, and the second nuclear pore membrane is used to filter the protective solution containing cells for detection, so as to retain the cells in the preservation solution on the first side of the second nuclear pore membrane. According to the detection requirements, immunoassay or FISH detection is directly performed on the first side of the second nuclear pore membrane. Compared with the existing detection process, there is no need to perform multiple preparation and multiple washing steps, and the detection is convenient and fast. The nuclear pore membrane biochip multi-detection device provided by the present invention performs morphological detection of exfoliated cells by absorbing the preservation solution enriched with cells, and uses the first side of the filtered second nuclear pore membrane to directly perform immunoassay or FISH detection, thereby achieving the purpose of simultaneously performing at least two detection methods, making the operation of lesion detection convenient and the accuracy higher. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0030] Figure 1 A schematic diagram of the structure of a detection device provided in an embodiment of the present invention;

[0031] Figure 2 A schematic diagram of the structure of a collection device provided in an embodiment of the present invention;

[0032] Figure 3 A top view of the first side of a second nuclear pore membrane provided by an embodiment of the present invention;

[0033] Figure 4 A side cross-sectional view of a second nuclear pore membrane provided by an embodiment of the present invention;

[0034] Figure 5 A schematic diagram of a method for transferring cells into a storage fluid cup provided in an embodiment of the present invention;

[0035] Among them, 10 is a collection device, 110 is a first nuclear pore membrane, 120 is a lower cylinder, 130 is an upper cylinder, 20 is a detection device, 210 is a preservation liquid cup, 220 is a preservation liquid, 230 is a second nuclear pore membrane, 240 is a cofferdam, 250 is a micropore area, and 260 is a reaction pool. DETAILED DESCRIPTION

[0036] The core of the present invention is to disclose a nuclear pore membrane biochip multi-link detection device to reduce the operational difficulty of lesion detection and improve the accuracy of lesion detection.

[0037] like Figure 1As shown, the nuclear pore membrane biochip multi-detection device provided by the present invention includes a collection device 10 for enriching cells for detection in human fluid, and a detection device 20 for performing the required lesion detection, wherein a first nuclear pore membrane 110 is provided in the collection device 10, and the first nuclear pore membrane 110 is used to enrich cells for detection in human fluid. When the body fluid flows through the collection device 10, it flows through the first nuclear pore membrane 110, and the first surface of the first nuclear pore membrane 110 faces the body fluid flowing through the first nuclear pore membrane 110. The first nuclear pore membrane 110 enriches the cells for detection in the body fluid on the first surface of the first nuclear pore membrane 110 through filtering. It should be noted that the parameters of the first nuclear pore membrane 110 are determined according to the size of the cells to be enriched. The detection device 20 includes a preservation liquid cup 210 containing a preservation liquid 220, and the preservation liquid 220 is used to protect the cell morphology. The opening of the preservation liquid cup 210 can allow the first surface of the first nuclear pore membrane 110 to be placed therein, so that after the cell enrichment is completed, the first surface of the first nuclear pore membrane 110 is inverted and immersed in the preservation liquid cup 210 to transfer the enriched cells. It should be noted that the purpose of transferring the enriched cells on the first surface of the first nuclear pore membrane 110 to the preservation liquid 220 is to protect the cell morphology and obtain a preservation liquid 220 with a high cell concentration. After the cell transfer is completed, the preservation liquid 220 is aspirated for staining and preparation for exfoliated cell morphology detection. Because there are more intact cells in the preservation liquid 220, the detection sensitivity is improved. It should be noted that the staining and preparation process is the same as the staining and preparation process in the prior art, and will not be repeated here. The cells in the preservation solution 220 are enriched through the filtering action of the first nuclear pore membrane 110, and the existing technology generally obtains enriched cells by centrifugation. Compared with centrifugation, filtration is less harmful to cells in body fluids, and a large amount of body fluids can be filtered to enrich more cells. Therefore, more intact cells can be obtained in the preservation solution 220. The first nuclear pore membrane 110 of the collection device 10 adopts a planar manner. Compared with the nuclear pore membrane in the form of a three-dimensional pointed cone, the collected cells will not be stuck in the seams of the membrane and in the gap between the membrane and the clamp, so it will not cause cell loss, thereby improving the sensitivity of cell morphology detection. The detection device 20 also includes a second nuclear pore membrane 230, which is detachably disposed at the bottom of the preservation liquid cup 210. The second nuclear pore membrane 230 is used to enrich the cells for detection in the preservation liquid 220 on the first side of the second nuclear pore membrane 230 through the filtering effect of the second nuclear pore membrane 230. After the second nuclear pore membrane 230 completes the filtering of the preservation liquid 220, according to the detection requirements, the second nuclear pore membrane 230 directly performs immunodetection or FISH detection on the first side of the second nuclear pore membrane 230. The detection methods include the following two:

[0038] The first detection method is direct immune response, the specific steps are as follows:

[0039] S1: Drop the cell lysis solution into the preservation solution cup 210. During the filtering process through the second nuclear pore membrane 230, the cells are lysed and the proteins in the cell nucleus are released. Since the second nuclear pore membrane 230 has been processed to enable it to directly capture antigen proteins, the proteins released from the cell nucleus are captured by the second nuclear pore membrane 230.

[0040] S2: blocked with bovine serum albumin;

[0041] S3: adding a labeled monoclonal antibody that can react with the antigen protein into the preservation liquid cup 210, and filtering the labeled antibody solution through the second nuclear pore membrane 230, at which time the nuclear antigen protein and the labeled monoclonal antibody are combined to form an immune reaction;

[0042] S4: adding the cleaning solution dropwise into the storage solution cup for percolation cleaning;

[0043] S5: The second nuclear pore membrane 230 is disassembled and readings are taken using a specific instrument, that is, the content of specific proteins in the nuclei of the enriched cells is detected.

[0044] The second detection method is double antibody sandwich immune reaction, the specific steps are:

[0045] S10: The second nuclear pore membrane 230 is pre-coated with a specific monoclonal antibody, called a capture antibody, and blocked with bovine serum albumin;

[0046] S20: installing the second nuclear pore membrane 230 in the detection device 20, and then enriching cells on the second nuclear pore membrane 230 according to the cell enrichment method provided by the present invention;

[0047] S30: adding a cell lysate to the preservation liquid cup 210 and filtering it through the second nuclear pore membrane 230, at which time the antigen protein enriched in the cell nucleus on the surface of the second nuclear pore membrane 230 is released and captured by the capture antibody coated on the second nuclear pore membrane 230;

[0048] S40: adding a labeled monoclonal antibody that can react with the antigen protein into the preservation liquid cup 210, and filtering the labeled antibody solution through the second nuclear pore membrane 230, so that the nuclear antigen protein and the labeled monoclonal antibody are combined to form an immune reaction;

[0049] S50: adding the cleaning solution dropwise into the storage solution cup 210 for percolation cleaning;

[0050] S60: Remove the second nuclear pore membrane 230 and read the data with a specific instrument, that is, detect the content of specific proteins in the nuclei of the enriched cells. Compared with the detection process of the existing immunoassay or FISH detection, the beneficial effects of the present invention are as follows:

[0051] The operation is simple, only a few times of sample addition and natural filtration are required to complete the whole process, making the detection convenient and fast; the filtration process makes the immune response efficient and the detection sensitivity high.

[0052] It should be noted that the parameters of the first nuclear pore membrane 110 and the second nuclear pore membrane 230 are determined according to the size of the cells to be enriched. In a specific embodiment of the present invention, it is necessary to enrich the tumor cells in urine for bladder cancer pathology detection. At this time, the pore size of the first nuclear pore membrane 110 is set to 2μm-11μm, and the pore density is set to 8E04 / cm 2 -3E06 / cm 2 The pore size of the second nuclear pore membrane 230 is set to 0.2 μm-10 μm, and the pore density is set to 1E05 / cm 2 -3E08 / cm 2 .

[0053] The nuclear pore membrane biochip multi-link detection device provided by the present invention enriches the cells for detection in the body fluid on the first surface of the first nuclear pore membrane 110 through the filtering effect of the first nuclear pore membrane 110. After the enrichment is completed, the first surface of the first nuclear pore membrane 110 is immersed in a preservation liquid cup 210 containing a preservation liquid 220 to transfer the cells on the first surface to the preservation liquid 220 of the detection device 20. Compared with the prior art that uses centrifugation to enrich cells in body fluids, the nuclear pore membrane biochip multi-link detection device provided by the present invention uses the first nuclear pore membrane 110 for enrichment and transfer, and the number of cells with intact morphology is larger. The preservation liquid 220 is absorbed for morphological detection of exfoliated cells, and the number of intact cells is larger, thereby improving the detection sensitivity and the detection accuracy; the detection device A second nuclear pore membrane 230 is detachably provided on the device 20, and the second nuclear pore membrane 230 is used to filter the protective solution containing cells for detection, so as to retain the cells in the preservation solution 220 on the first side of the second nuclear pore membrane 230. According to the detection requirements, immunoassay or FISH detection is directly performed on the first side of the second nuclear pore membrane 230. Compared with the existing detection process, the detection is convenient and quick. The nuclear pore membrane biochip multi-detection device provided by the present invention performs morphological detection of exfoliated cells by absorbing the preservation solution 220 enriched with cells, and uses the first side of the filtered second nuclear pore membrane 230 for immunoassay or FISH detection, thereby achieving the purpose of simultaneously performing cell morphology and immunoassay of at least two proteins, making the operation of lesion detection convenient and more accurate.

[0054] In order to further improve the accuracy of lesion detection, Figure 2 and Figure 3As shown, in the nuclear pore membrane biochip multi-detection device provided by the present invention, the first side of the second nuclear pore membrane 230 is divided into at least two micropore areas 250 by the cofferdam 240. It should be noted that the first side of the second nuclear pore membrane 230 is a side for retaining cells for detection in the preservation solution 220, and the cofferdam 240 is a protective structure higher than the micropore area 250. The existence of the cofferdam 240 allows the cells in the preservation solution 220 to be retained in the micropore area 250. At least two micropore areas 250 can meet the requirements of simultaneously performing immunodetection and / or FISH molecular detection of different proteins on the upper surface of the second nuclear pore membrane 230, and different tumor markers are respectively dripped on different micropore areas 250 to perform two or more types of detection to improve the accuracy of lesion detection.

[0055] It should be noted that the first side of the second nuclear pore membrane 230 is divided into at least two micropore areas 250 by the cofferdam 240, so that different micropore areas 250 on the second nuclear pore membrane 230 can be coated with different monoclonal antibodies. By adopting the above-mentioned operation process, the detection of different antigen proteins in the cell can be completed at the same time, and the purpose of simultaneously performing at least two methods of detection is achieved, so that the lesion detection meets the characteristics of high specificity and high sensitivity, and avoids the problem of false negatives in lesion detection; the device provided by the present invention can simultaneously realize multiple detections such as body fluid cell enrichment, cell morphology detection, cell-specific protein immune detection, FISH molecular detection, etc., so that the accuracy of lesion detection is higher and the operation is more convenient.

[0056] It should be noted that the number of micropore areas 250 can be set according to detection requirements, such as Figure 3 and Figure 4 As shown, in a specific embodiment of the present invention, four microporous areas 250 are provided to simultaneously perform immunoassays such as one or more of NMP22 (urine nuclear matrix protein 22), CK20 (cytokeratin 20), BTA (bladder tumor antigen), and FISH.

[0057] Furthermore, if Figure 1As shown, the second nuclear pore membrane 230 needs to be tested for lesions after filtering the preservation solution 220. In order to facilitate the movement of the second nuclear pore membrane 230, the second nuclear pore membrane 230 is detachably mounted on the bottom opening of the preservation solution cup 210 through the reaction pool 260. The bottom of the reaction pool 260 can be opened to discharge the preservation solution 220 in the preservation solution cup 210. The second nuclear pore membrane 230 is fixedly connected to the top of the reaction pool 260. The first side of the second nuclear pore membrane 230 faces the preservation solution 220. When filtering the preservation solution 220 is required, the bottom of the reaction pool 260 is opened so that the preservation solution 220 flows out of the preservation solution cup 210 through the reaction pool 260 and flows through the second nuclear pore membrane 230 to filter the preservation solution 220. After the filtration is completed, an immune reaction or FISH fluorescence in situ hybridization reaction can be performed on the side of the second nuclear pore membrane 230 where cells are enriched. After the reaction, the second nuclear pore membrane is removed, and qualitative and quantitative detection can be performed with an instrument, which is convenient and quick.

[0058] Furthermore, in a specific embodiment of the present invention, the reaction pool 260 is threadedly connected to the bottom of the preservation liquid cup 210 and a seal is provided on the outside to achieve the detachable reaction pool 260 while ensuring the sealing with the preservation liquid cup 210, and the second nuclear pore membrane 230 is fixedly connected to the top of the reaction pool 260 by hot melt welding.

[0059] Furthermore, if Figure 2 As shown, in the nuclear pore membrane biochip multi-detection device provided by the present invention, the first nuclear pore membrane 110 is fixedly connected to the first side of the lower cylinder 120 to achieve the purpose of enriching the body fluid for detecting cells. The body fluid flows through the lower cylinder 120 and is filtered through the first nuclear pore membrane 110. The first surface of the first nuclear pore membrane 110 faces the direction of the body fluid flowing through the first nuclear pore membrane 110. After the first surface of the first nuclear pore membrane 110 completes the enrichment of the cells for detection in the body fluid, the lower cylinder 120 is moved and flipped to immerse the first surface of the first nuclear pore membrane 110 into the preservation liquid cup 210 for cell transfer.

[0060] Furthermore, in a specific embodiment of the present invention, a truncated cone-shaped flow channel is provided inside the lower cylinder 120. When the collection device 10 is enriching cells for detection in body fluids, the smaller side of the truncated cone-shaped flow channel faces downward, so that the body fluid flowing through the lower cylinder 120 flows out of the lower cylinder 120 from the smaller side of the cone without sticking to the side wall of the lower cylinder 120 or flowing out along the side wall of the lower cylinder 120 to contaminate the operator.

[0061] Furthermore, if Figure 5As shown, in order to make the process of transferring cells more convenient, in a specific embodiment of the present invention, the outer surface of the first side of the lower cylinder 120 is provided with an external thread, and the top of the preservation liquid cup 210 is provided with an internal thread to be threadedly connected with the first side of the lower cylinder 120, so that the lower cylinder 120 can be fixedly connected to the preservation liquid cup 210 and ensure that the first surface of the first nuclear pore membrane 110 is completely immersed in the preservation liquid 220. At the same time, the lower cylinder 120 can be fixedly connected to the preservation liquid cup 210, and the cell shedding rate on the first surface of the first nuclear pore membrane 110 can be accelerated by oscillating. The fixedly connected lower cylinder 120 and the preservation liquid cup 210 are used to prevent the preservation liquid 220 from overflowing from the preservation liquid cup 210 during the oscillation process.

[0062] It should be noted that in order to facilitate the shedding of cells on the first nuclear pore membrane 110 , when the lower cylinder 120 is inverted and threadedly connected to the preservation liquid cup 210 , the preservation liquid 220 is poured into the lower cylinder 120 to flush the first nuclear pore membrane 110 to accelerate the shedding of cells from the first surface of the first nuclear pore membrane 110 .

[0063] Furthermore, if Figure 2 As shown, in another specific embodiment of the present invention, the collection device 10 also includes an upper cylinder 130, and one side of the upper cylinder 130 is threadedly matched with the first side of the lower cylinder 120. After the upper cylinder 130 and the lower cylinder 120 are threadedly connected, the body fluid is filtered, which can keep the body fluid inside the upper cylinder 130 and slowly pass through the first nuclear pore membrane 110 for filtration. Compared with the body fluid directly flowing through the first nuclear pore membrane 110, the filtration effect is better and the body fluid can be prevented from overflowing from the surroundings of the first nuclear pore membrane 110.

[0064] Furthermore, in the nuclear pore membrane biochip multiplex detection device provided by the present invention, the dam 240 is a high molecular polymer layer deposited on the first side of the second nuclear pore membrane 230 by printing.

[0065] Furthermore, the first nuclear pore membrane 110 is fixed to one side of the lower cylinder 120 by heat-melt welding or adhesive. Here, it is preferred that the first nuclear pore membrane 110 is fixedly connected to one side of the lower cylinder 120 by heat-melt welding.

[0066] The present invention also provides a method for using a nuclear pore membrane biochip multi-link detection device to detect lesions, using a nuclear pore membrane biochip multi-link detection device having one or more of the above technical effects to detect lesions. The method provided by the present invention can achieve the purpose of using the nuclear pore membrane biochip multi-link detection device to perform at least two methods of detection. The method provided by the present invention at least includes the following steps:

[0067] S01: enriching body fluid cells, using the collection device 10 to enrich the cells in the body fluid on the first surface of the first nuclear pore membrane 110;

[0068] S02: Transferring cells, turning over the collection device 10 so that the first surface of the first nuclear pore membrane 110 faces the opening of the preservation liquid cup 210, and immersing the first surface of the first nuclear pore membrane 110 into the preservation liquid 220 in the preservation liquid cup 210, and transferring the cells on the first surface to the preservation liquid 220 in the preservation liquid cup 210 for morphological detection of exfoliated cells;

[0069] It should be noted that, in step S02, Figure 5 As shown, the collection device 10 is turned over and the cells are transferred.

[0070] S03: enriching the cells in the preservation solution 220, and enriching the cells in the preservation solution 220 to the first side of the second nuclear pore membrane 230 through the filtering effect of the second nuclear pore membrane 230, so as to be used for immunodetection or FISH detection.

[0071] Furthermore, in the method of use provided by this method, step S02 specifically includes:

[0072] S21: turning over the collection device 10 so that the first surface of the first nuclear pore membrane 110 faces the opening of the preservation liquid cup 210, and immersing the first surface of the first nuclear pore membrane 110 into the preservation liquid 220 in the preservation liquid cup 210;

[0073] S22 : Using an oscillator to oscillate the preservation solution cup 210 to transfer the cells on the first surface of the first nucleopore membrane 110 into the preservation solution 220 .

[0074] The terms "first" and "second" and the like in the specification and claims of the present invention and the above drawings are used to distinguish different objects, rather than to describe a specific order. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device including a series of steps or units is not limited to the listed steps or units, but may include steps or units that are not listed.

[0075] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A nuclear pore membrane biochip multi-link detection device, It is characterized in that The invention comprises a collection device (10) and a detection device (20), wherein the collection device (10) comprises a first nuclear pore membrane (110), wherein a first surface of the first nuclear pore membrane (110) faces a body fluid flowing through the first nuclear pore membrane (110), so as to enrich cells for detection in the body fluid on the first surface; The first nuclear pore membrane (110) is in a planar manner; The detection device (20) comprises: a preservation liquid cup (210) containing a preservation liquid (220), wherein the first surface can be immersed in the preservation liquid cup (210), and the preservation liquid (220) is used to protect the cell morphology; a second nuclear pore membrane (230), the second nuclear pore membrane (230) being detachably disposed at the bottom of the preservation solution cup (210), the second nuclear pore membrane (230) being used to filter the preservation solution (220) so as to enrich the cells for detection in the preservation solution (220) on the first side of the second nuclear pore membrane (230); The second nuclear pore membrane (230) is detachably arranged at the bottom of the preservation liquid cup (210) through the reaction pool (260), the reaction pool (260) is detachably installed at the bottom opening of the preservation liquid cup (210), and the bottom of the reaction pool (260) can be opened to discharge the preservation liquid (220) in the preservation liquid cup (210), and the second nuclear pore membrane (230) is fixedly connected to the top of the reaction pool (260).

2. The nuclear pore membrane biochip multi-link detection device as claimed in claim 1, It is characterized in that The first side of the second nuclear pore membrane (230) is divided into at least two micropore areas (250) by a dam (240) to separate the cells enriched on the first side of the second nuclear pore membrane (230) into at least two areas for performing different types of detection.

3. The nuclear pore membrane biochip multi-link detection device as claimed in claim 1, It is characterized in that The first nuclear pore membrane (110) is fixedly connected to the first side of the lower cylinder (120) to enrich the cells for detection in the body fluid flowing through the lower cylinder (120).

4. The nuclear pore membrane biochip multi-link detection device as claimed in claim 3, It is characterized in that A truncated cone-shaped flow channel is provided inside the lower cylinder (120), so that the body fluid flowing through the lower cylinder (120) flows out of the lower cylinder (120) from the smaller side of the cone.

5. The nuclear pore membrane biochip multi-link detection device as claimed in claim 3, It is characterized in that An outer surface of a first side of the lower cylinder (120) is provided with an external thread, and a top of the preservation liquid cup (210) is provided with an internal thread for threaded connection with the lower cylinder (120).

6. The nuclear pore membrane biochip multi-link detection device as claimed in claim 5, It is characterized in that The collecting device (10) further comprises an upper cylinder (130), one side of which is provided with an internal thread that matches with the first side thread of the lower cylinder (120).

7. The nuclear pore membrane biochip multi-link detection device as claimed in claim 2, It is characterized in that The dam (240) is a high molecular polymer layer printed and deposited on the first side of the second nuclear pore membrane (230).

8. The nuclear pore membrane biochip multi-link detection device as claimed in claim 1, It is characterized in that The reaction pool (260) is threadedly connected to the bottom opening of the preservation liquid cup (210).

9. The nuclear pore membrane biochip multi-link detection device as claimed in claim 3, It is characterized in that The first nuclear pore membrane (110) is fixed to one side of the lower cylinder (120) by hot-melt welding or adhesive.

10. A method for using a nuclear pore membrane biochip multi-link detection device, It is characterized in that The multiplex detection is performed using the nuclear pore membrane biochip multiplex detection device provided by any one of claims 1 to 9, wherein the method of use comprises at least the following steps: Enriching body fluid cells: using the collection device (10) to enrich cells in the body fluid on the first surface of the first nuclear pore membrane (110); Transferring cells: turning over the collection device (10) so that the first surface of the first nuclear pore membrane (110) faces the opening of the preservation liquid cup (210), and immersing the first surface of the first nuclear pore membrane (110) in the preservation liquid (220) in the preservation liquid cup (210), and transferring the cells on the first surface to the preservation liquid (220) in the preservation liquid cup (210) for morphological detection of exfoliated cells; Enrichment of preservation fluid cells: The cells remaining in the preservation fluid (220) after cell morphology detection are enriched to the first side of the second nuclear pore membrane (230) through the filtering effect of the second nuclear pore membrane (230) for use in immunodetection or FISH molecular detection.

11. The method of use according to claim 10, It is characterized in that The cell transfer step specifically includes: Turning over the collection device (10) so that the first surface of the first nuclear pore membrane (110) faces the opening of the preservation liquid cup (210), and immersing the first surface of the first nuclear pore membrane (110) in the preservation liquid (220) in the preservation liquid cup (210); The preservation solution cup (210) is shaken using an oscillator to transfer the cells on the first surface of the first nuclear pore membrane (110) into the preservation solution (220).

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