An integrated microwave-microfluidic chip and its preparation and detection method

Abstract

The present invention relates to the field of circulating tumor cell detection technology, and provides an integrated microwave-microfluidic chip, its preparation method, and detection method. The chip comprises a copper-based glass slide, a microwave sensor, and a microfluidic chip arranged from bottom to top. The microwave sensor and the microfluidic chip are both provided with protein-labeled regions, and the protein-labeled regions on the microwave sensor and the microfluidic chip are aligned. The microfluidic chip is provided with a microfluidic channel, and the central axis of the protein-labeled region is aligned with the central axis of the microfluidic channel. The protein-labeled regions of the microfluidic chip and the microwave sensor are both labeled with specific antibodies, and the specific antibodies are used to capture biomarkers in blood samples. This method achieves label-free and non-invasive detection of circulating tumor cells, avoiding the shortcomings of traditional methods such as the need for precision instruments, long time consumption, high cost, and single detection.

Description

Technical Field

[0001] The present invention relates to the technical field of circulating tumor cell detection, and in particular to an integrated microwave-microfluidic chip and a preparation method and a detection method thereof. Background Art

[0002] The statements in this section merely provide background art related to the present invention and do not necessarily constitute prior art.

[0003] Circulating tumor cells (CTCs) are tumor cells that circulate in the body, detaching from the primary tumor and entering the blood or lymphatic system. Although most shed cells can undergo apoptosis or be phagocytosed, a small number of CTCs spread to other parts of the body and form new tumors. CTCs are one of the important factors in tumor metastasis. In the early stages of cancer, CTC detection can help doctors detect the presence of circulating tumor cells in the early stages of cancer, thereby increasing the chance of successful treatment and reducing patient mortality. In addition, detecting CTCs helps determine the type, stage, and severity of cancer, thereby formulating a more accurate treatment plan. During cancer treatment, monitoring the number and nature of CTCs can help doctors evaluate the effectiveness of treatment. Therefore, accurate and rapid detection of the number of circulating tumor cells can help in early screening and treatment of cancer.

[0004] Microfluidics integrates sample separation, enrichment, reaction, and detection functions into a single chip, enabling automated testing of trace samples. It boasts the advantages of small size, rapid analysis, high automation, low cost, and minimal capital requirements. In medical testing, it effectively avoids the time-consuming and contaminating nature of traditional methods. However, traditional microfluidic chips rely on precision equipment such as fluorescence microscopes, fluorescence spectrophotometers, or microplate readers.

[0005] Current CTCs detection technology often uses a combination of microfluidics and immunofluorescence staining. The immunofluorescence staining method mainly includes the following steps: sample collection, cell separation, cell fixation, multiple washes, and image analysis. Its principle is to use fluorescently labeled antibodies to specifically bind to specific proteins on CTCs, and then use instruments such as fluorescence microscopes, flow cytometers, or microplate readers to detect and observe CTCs. Immunofluorescence staining for CTCs is a highly specific and sensitive method, but it has disadvantages such as complex sample processing, high cost, limited sensitivity, and low degree of automation. This will face difficulties when processing a large number of samples. Specifically:

[0006] (1) Difficulty in separation: CTCs are very rare in the blood, usually only a few per milliliter of blood, and are mixed with red blood cells and white blood cells. Immunofluorescence detection usually requires centrifugation to separate plasma, white blood cells, and red blood cells, but this process is cumbersome and time-consuming.

[0007] (2) Fluorescent labeling damage: Immunofluorescence detection uses antibodies labeled with fluorescent groups to specifically bind to proteins on CTCs; however, CTCs are rare, and fluorescent labeling may cause irreversible damage to CTCs, which may affect subsequent analysis of CTCs;

[0008] (3) High cost and lack of portability: Image analysis usually requires the use of expensive equipment, such as flow cytometers or fluorescence microscopes, which increases costs. In addition, these devices are usually not easy to carry and require professional technicians to operate. They have limited applicability and are not suitable for instant detection. Moreover, the report usually takes 1-2 days to be issued. In addition, during the observation process, the light intensity of the fluorescent group will gradually weaken over time, requiring timely observation and analysis.

[0009] (4) Low degree of automation and human errors: Steps such as sample collection and processing usually require manual operation and have a low degree of automation; this may lead to the introduction of human errors and affect the accuracy of detection.

[0010] (5) Single detection: Existing microwave detection technology can only realize sensing detection in a single area. Dual sensing and multi-area detection are prone to mutual influence, resulting in a decrease in sensing accuracy. Summary of the Invention

[0011] In order to address the shortcomings of the existing technology, the present invention provides an integrated microwave-microfluidic chip and its preparation method and detection method. By combining microwave detection technology and microfluidic technology, label-free and non-invasive detection of circulating tumor cells is achieved, avoiding the shortcomings of traditional methods such as the need for precision instruments, long time consumption, high cost, and single detection.

[0012] In order to achieve the above object, the present invention adopts the following technical solutions:

[0013] A first aspect of the present invention provides an integrated microwave-microfluidic chip.

[0014] An integrated microwave-microfluidic chip includes a copper-based glass slide, a microwave sensor, and a microfluidic chip arranged from bottom to top;

[0015] The microwave sensor and the microfluidic chip are both provided with a protein labeling region, and the protein labeling regions on the microwave sensor and the microfluidic chip are aligned; a microfluidic channel is provided on the microfluidic chip, and the central axis of the protein labeling region is aligned with the central axis of the microfluidic channel;

[0016] The protein labeling areas of the microfluidic chip and the microwave sensor are both labeled with specific antibodies, and the specific antibodies are used to capture biomarkers in blood samples.

[0017] Furthermore, a blood sample inlet is provided at one end of the microfluidic channel, and a blood sample outlet is provided at the other end.

[0018] Furthermore, the microwave sensor includes a forked microstrip line and two square inner rings with openings, and the two square inner rings with openings are both arranged between two forks of the forked microstrip line.

[0019] Furthermore, protein labeling regions are provided at the openings of the two square inner rings, and the two protein labeling regions have the same shape but different sizes.

[0020] Furthermore, the forked microstrip line is connected to a data connection port, and the data connection port is used to connect to a host computer.

[0021] A second aspect of the present invention provides a method for preparing an integrated microwave-microfluidic chip as described in the first aspect, comprising the following steps:

[0022] After setting a microwave sensor on a copper-based glass slide, the copper-based glass slide is cleaned and salted, and after marking the microwave sensor with an activated antibody solution, the antibody-coated microwave sensor is washed with deionized water and air-dried;

[0023] After cleaning and salinizing the microfluidic chip, the activated antibody solution is labeled onto the microfluidic chip, and the antibody-coated microfluidic chip is washed with deionized water and air-dried;

[0024] The microfluidic channel of the microfluidic chip and the protein labeling area on the microwave sensor are aligned and then bonded.

[0025] Furthermore, the method further includes welding the data connection port to the microwave sensor.

[0026] Furthermore, the manufacturing step of the microfluidic chip further includes: punching holes at both ends of the microfluidic channel of the microfluidic chip to obtain a blood sample inlet and a blood sample outlet.

[0027] A third aspect of the present invention provides a detection method using an integrated microwave-microfluidic chip as described in the first aspect, comprising the following steps:

[0028] Transmitting a signal to a microwave sensor;

[0029] The blood sample enters the microfluidic channel and flows through the protein labeling area. The specific antibodies in the protein labeling area capture the biomarkers in the blood.

[0030] When the biomarker is captured by the specific antibody, it causes a signal change.

[0031] Furthermore, the signal change amount is linearly related to the biomarker capture amount.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] 1. The integrated microwave-microfluidic chip described in the present invention realizes label-free detection: a microwave resonator combined with a microfluidic chip is used to achieve label-free detection, effectively avoiding damage to the sample caused by markers such as fluorescent groups, maintaining the integrity of the sample, which provides a guarantee for further analysis.

[0034] 2. The integrated microwave-microfluidic chip described in the present invention realizes automatic sampling: it utilizes the capillary phenomenon of the microfluidic channel to realize automatic sampling, greatly reducing the manual operation process and avoiding errors caused by human intervention.

[0035] 3. The integrated microwave-microfluidic chip described in the present invention achieves miniaturization and integration: the microfluidic chip and microwave sensor allow multiple operation steps to be performed in a tiny chip, thereby greatly shortening the operation time and reducing the volume.

[0036] 4. The integrated microwave-microfluidic chip described in the present invention achieves rapid detection: a microwave resonator combined with a microfluidic chip is used to achieve label-free detection, avoiding the preparation and processing steps of markers, thereby reducing the complexity of the experiment; when a blood sample flows into the microfluidic channel, CTCs will bind to specific antibodies and be fixed in a specific area, while white blood cells and red blood cells cannot be captured due to the lack of corresponding antibodies, simplifying the steps of CTC separation, capture and enrichment, and shortening the operation time.

[0037] 5. The integrated microwave-microfluidic chip described in the present invention achieves low cost: the microfluidic chip and microwave sensor are simple to manufacture and inexpensive; at the same time, the combination of the two avoids the use of large instruments and shortens the operation steps, saving costs.

[0038] 6. The integrated microwave-microfluidic chip described in the present invention realizes dual-region collaborative detection, avoiding the disadvantage of single detection of traditional methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments and descriptions of the present invention are used to explain the present invention and do not constitute an improper limitation to the present invention.

[0040] Figure 1 A perspective view of an integrated microwave-microfluidic chip according to Example 1 of the present invention;

[0041] Figure 2 This is a structural diagram of a microwave sensor according to Example 1 of the present invention;

[0042] Figure 3 3D structural diagram of the integrated microwave-microfluidic chip according to Example 1 of the present invention;

[0043] Figure 4 This is a schematic diagram of the integrated microwave-microfluidic chip according to Example 1 of the present invention;

[0044] Figure 5 This is a dual-signal frequency distribution diagram of Example 1 of the present invention;

[0045] Figure 6 This is a mask diagram of the microwave sensor of Example 1 of the present invention. DETAILED DESCRIPTION

[0046] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0047] It should be noted that the following detailed descriptions are exemplary and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0048] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0049] In the present invention, terms such as "upper", "lower", "left", "right", "front", "back", "vertical", "horizontal", "side", "bottom", etc. indicating directions or positional relationships are based on the directions or positional relationships shown in the accompanying drawings. They are relational words determined only for the convenience of describing the structural relationships of the various parts or elements of the present invention, and do not specifically refer to any part or element in the present invention, and should not be understood as limiting the present invention.

[0050] In the present invention, terms such as "fixed connection," "connected," and "connection" should be interpreted broadly to mean a fixed connection, an integral connection, or a detachable connection; a direct connection or an indirect connection through an intermediary. Relevant researchers or technicians in this field may determine the specific meanings of these terms in the present invention based on specific circumstances, and they should not be construed as limitations of the present invention.

[0051] In the absence of conflict, the embodiments of the present invention and the features thereof may be combined with each other.

[0052] Explanation of terms:

[0053] Microstrip line: A microwave transmission line consisting of a single conductor strip supported on a dielectric substrate.

[0054] Example 1

[0055] Embodiment 1 of the present invention provides an integrated microwave-microfluidic chip.

[0056] The principle of microwave detection is based on the field disturbance caused by the sample being tested, which leads to changes in the resonant frequency and notch amplitude. Different types of tissues and body fluids have different components, which leads to differences in dielectric properties. By measuring the differences in dielectric properties, different samples can be identified and detected. Different types of tissues and body fluids have different dielectric properties, which leads to differences in dielectric properties. By measuring the differences in dielectric properties, different samples can be identified and detected. Microwave detection devices can directly detect the dielectric properties of samples and have the advantages of high sensitivity, real-time response, and simple operation. With the continuous development of microwave technology, many types of microwave sensors have been developed, among which microwave resonator sensors are one type. They can convert the dielectric properties of the sample to be tested into electrical signals, thereby measuring the resonant frequency or relative oscillation phase of the parameters. They have the advantages of high integration, high sensitivity, non-invasiveness, real-time response, and low cost.

[0057] The integrated microwave-microfluidic chip provided in this embodiment greatly reduces the size by combining microwave detection technology and microfluidic technology, and realizes label-free and non-invasive detection of circulating tumor cells, avoiding the shortcomings of traditional methods such as the need for precise instruments, long time consumption, high cost, and single detection.

[0058] like Figure 1 、 Figure 2 and Figure 3 As shown, this embodiment provides an integrated microwave-microfluidic chip, including a data connection port 6 and a copper-based glass slide 1, a microwave sensor (also known as a microwave resonator) 4 and a PDMS chip (also known as a microfluidic chip) 5 arranged from bottom to top.

[0059] The microwave sensor 4 is arranged on the copper-based glass slide 1 , and the PDMS chip 5 is arranged on the microwave sensor 4 .

[0060] like Figure 2 As shown, the microwave sensor 4 includes a forked microstrip line and two square inner rings with openings, and the two square inner rings with openings are both arranged between the two forks of the forked microstrip line; the two square inner rings have the same shape but inconsistent size; the opening sizes of the two square inner rings are inconsistent; the first protein marking area 8 and the second protein marking area 9 are respectively arranged at the openings of the two square inner rings, and the first protein marking area 8 and the second protein marking area 9 have the same shape but inconsistent size, and the forked microstrip line (fork handle) is connected to the data connection port 6.

[0061] A microfluidic channel 3 is provided at the bottom of the PDMS chip 5. A blood sample inlet 2 is provided at one end of the microfluidic channel 3, and a blood sample outlet 7 is provided at the other end.

[0062] After the central axes of the first protein labeling region 8 and the second protein labeling region 9 are aligned with the central axis of the microfluidic channel 3 , the copper-based glass slide 1 and the PDMS chip 5 are bonded.

[0063] Wherein, at the positions of the first protein labeling region 8 and the second protein labeling region 9 , the PDMS chip 5 and the microwave sensor 4 are both labeled with specific antibodies 10 , which are used to capture biomarkers in the blood sample.

[0064] In this embodiment, the specific antibody 10 is an antibody that matches CTCs. If the specific antibody type is changed, it can be used to detect viruses, bacteria, or various biomarkers. The first protein labeling region 8 and the second protein labeling region 9 can be labeled with the same type of antibody or different types of antibodies. If different types of antibodies are used, it can be used to simultaneously detect two CTCs or biomarkers.

[0065] like Figure 5 As shown, the signal of the microwave sensor 4 has characteristic signals at different frequencies, that is, the frequencies of the characteristic signals corresponding to the two protein marker regions may be different.

[0066] Among them, the first protein labeling region 8 corresponds to a characteristic signal of 5.29 GHz, and the second protein labeling region 9 corresponds to a characteristic signal of 7.67 GHz, and the signals do not affect each other.

[0067] In this embodiment, the first protein-labeled region 8 and the second protein-labeled region 9 can be labeled with the same or different types of antibodies. If they are labeled with different types of antibodies, they can be used to simultaneously detect two CTCs or biomarkers. If they are labeled with the same antibody, the second protein-labeled region 9 can capture the analyte not captured by the first protein-labeled region 8, thereby improving detection sensitivity.

[0068] In this embodiment, the signal frequency is associated with parameters such as the structure, size, and opening size of the microwave sensor 4. If the parameters such as the structure, size, and opening size of the microwave sensor 4 are changed, the characteristic signal will shift.

[0069] The microwave sensor is connected to a data connection port 6 for connecting to a host computer, which may be a vector network analyzer.

[0070] This embodiment provides an integrated microwave-microfluidic chip, such as Figure 4 As shown, its working principle is as follows: the vector network analysis module integrated with WiFI and Bluetooth modules transmits signals to the microwave sensor in real time through the data connection port 6 and receives feedback signals. The blood sample enters the microfluidic channel 3 from the entrance of the sample inlet 2 and flows through the first protein labeling area 8 and the second protein labeling area 9. The specific antibodies 10 in the first protein labeling area 8 and the second protein labeling area 9 capture the circulating tumor cells (CTCs) 11 in the blood. When the circulating tumor cells (CTCs) 11 are captured by the specific antibodies 10, it will cause the feedback signal to change. The change in the feedback signal is linearly related to the capture amount of the circulating tumor cells (CTCs) 11.

[0071] The blood contains circulating tumor cells (CTCs) 11 , white blood cells 12 , and red blood cells 13 .

[0072] The present embodiment provides an integrated microwave-microfluidic chip, which achieves the capture and enrichment of CTCs in specific areas by fixing specific proteins related to CTCs in specific areas of the microfluidic chip. When the blood sample flows into the microfluidic channel, CTCs will bind to specific antibodies and be fixed in a specific area, while white blood cells and red blood cells cannot be captured due to the lack of corresponding antibodies, which simplifies the separation and enrichment steps of CTCs and improves the accuracy and sensitivity of detection. A microwave resonator combined with a microfluidic chip is used to achieve label-free detection, thereby avoiding the use of fluorescent microscopes or other fluorescent equipment with high costs and poor portability, avoiding damage to the sample by markers such as fluorescent groups, and ensuring the integrity of the sample. The test results are obtained through a vector network analyzer integrated with WiFi and Bluetooth modules, and directly transmitted to a mobile phone or computer for data analysis.

[0073] The integrated microwave-microfluidic chip provided in this embodiment has the following advantages:

[0074] (1) Label-free detection: A microwave resonator combined with a microfluidic chip is used to achieve label-free detection, effectively avoiding damage to the sample caused by markers such as fluorescent groups and maintaining the integrity of the sample, which provides a guarantee for further analysis;

[0075] (2) Automatic injection: Utilizing the capillary phenomenon of the microfluidic channel, automatic injection is achieved, which greatly reduces the manual operation process and avoids errors caused by manual operation;

[0076] (3) Miniaturization and integration: Microfluidic chips and microwave sensors allow multiple operation steps to be performed in a tiny chip, thus greatly shortening the operation time and reducing the volume;

[0077] (4) Rapid detection: A microwave resonator combined with a microfluidic chip is used to achieve label-free detection, avoiding the preparation and processing steps of the marker, thereby reducing the complexity of the experiment; when the blood sample flows into the microfluidic channel, CTCs will bind to specific antibodies and be fixed in a specific area, while white blood cells and red blood cells cannot be captured due to the lack of corresponding antibodies, simplifying the steps of CTC separation, capture and enrichment, and shortening the operation time;

[0078] (5) Low cost: Microfluidic chips and microwave sensors are simple to make and inexpensive; at the same time, the combination of the two avoids the use of large instruments and shortens the operation steps, saving costs.

[0079] (6) Dual-area collaborative detection can be achieved.

[0080] Example 2

[0081] Example 2 of the present invention provides a method for preparing an integrated microwave-microfluidic chip as in Example 1, comprising the following steps:

[0082] Step 1: The steps for making a microwave sensor are as follows:

[0083] (101) After the photoresist is evenly spin-coated (fully coated) onto a copper-plated glass slide (also known as a copper-based glass slide) 1, it is baked.

[0084] (102) A pattern of a microwave sensor is formed on a copper-plated glass slide (hereinafter referred to as a slide) 1 with a mask by ultraviolet exposure. The shape of the mask is as follows Figure 6As shown, it includes a fork mask and two square inner ring masks with openings, and the two square inner ring masks with openings are both arranged between the two forks of the fork mask; the two square inner ring masks have the same shape but inconsistent sizes; the opening sizes of the two square inner ring masks are inconsistent.

[0085] (103) After the pattern of the microwave sensor on the glass slide is developed using a negative photoresist developer, the glass slide is immersed in a plating solution for electroplating.

[0086] (104) After electroplating, the photoresist and copper base are removed, leaving the microwave sensor 4 on the glass slide.

[0087] (105) The glass slide was cleaned and placed in APTES (γ-aminopropyltriethoxysilane) for salting; 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide solution, and antibody solution matching CTCs were fully mixed to obtain an activated antibody solution (containing specific antibody 10), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide solution were used to activate the antibody.

[0088] (106) The activated antibody solution is labeled onto the first protein labeling area 8 and the second protein labeling area 9 on the microwave sensor 4; the antibody-coated microwave sensor is then washed with deionized water and air-dried.

[0089] Step 2: The steps for making the microfluidic chip are as follows:

[0090] (201) Polydimethylsiloxane (PDMS) and the curing agent were mixed evenly and degassed in a vacuum box to obtain a mixed solution for later use.

[0091] (202) An appropriate amount of the mixed solution is introduced into a mold prepared by soft lithography. After curing, a PDMS chip is obtained. A hole is punched at an appropriate position of the microfluidic channel 3 of the PDMS chip. One end of the microfluidic channel 3 is a blood sample inlet 2, and the other end is a blood sample outlet 7.

[0092] (203) The PDMS chip was cleaned and placed in APTES for salting; 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide solution, and antibody solution matching CTCs were fully mixed to obtain an activated antibody solution, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide solution were used to activate the antibody.

[0093] (204) The activated antibody solution was labeled onto the first protein labeling area 8 and the second protein labeling area 9 of the PDMS chip, and then the antibody-coated PDMS chip was washed with deionized water and air-dried.

[0094] Step 3: Align the microfluidic channel 3 of the PDMS chip and the microwave sensor that has been labeled with the antibody (ie, align the microfluidic channel 3 of the PDMS chip and the first protein labeling area 8 and the second protein labeling area 9 on the microwave sensor 4) and then bond them.

[0095] Step 4: Weld the signal transmission port (ie, data connection port) 6 to the microwave sensor.

[0096] Step 5: Connect the microwave sensor to the vector network analysis module integrated with Wi-Fi and Bluetooth modules to transmit the signal to the mobile phone or computer.

[0097] Example 3

[0098] Example 3 of the present invention provides a detection method based on an integrated microwave-microfluidic chip as in Example 1, which can achieve dual-region collaborative detection, including the following steps:

[0099] Step 1: The vector network analysis module integrated with WiFI and Bluetooth modules transmits signals to the microwave sensor through the data connection port 6 in real time and receives feedback signals;

[0100] Step 2: The blood sample enters the microfluidic channel 3 from the sample inlet 2 and flows through the first protein labeling area 8 and the second protein labeling area 9 in sequence. The specific antibodies 10 in the first protein labeling area 8 and the second protein labeling area 9 capture circulating tumor cells (CTCs) 11 in the blood.

[0101] Step 3: When circulating tumor cells (CTCs) 11 are captured by the specific antibody 10, the feedback signal will change, and the amount of change in the feedback signal is linearly related to the amount of circulating tumor cells (CTCs) 11 captured.

[0102] In this embodiment, the first protein labeling region 8 corresponds to a characteristic signal (characteristic peak) of 5.29 GHz, such as Figure 5 As shown, when the sample is only loaded on the first protein labeling area 8, the 5.29 GHz signal will change (the characteristic peak will move); the second protein labeling area 9 corresponds to the 7.67 GHz characteristic signal, and the signals do not affect each other.

[0103] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. An integrated microwave-microfluidic chip, characterized in that: It includes a copper-based glass slide, microwave sensor and microfluidic chip arranged from bottom to top; The microwave sensor and the microfluidic chip are both provided with a protein labeling region, and the protein labeling regions on the microwave sensor and the microfluidic chip are aligned; a microfluidic channel is provided on the microfluidic chip, and the central axis of the protein labeling region is aligned with the central axis of the microfluidic channel; The protein labeling areas of the microfluidic chip and the microwave sensor are both labeled with specific antibodies, and the specific antibodies are used to capture biomarkers in blood samples; The microwave sensor includes a forked microstrip line and two square inner rings with openings, and the two square inner rings with openings are both arranged between the two forks of the forked microstrip line; protein marking areas are both provided at the openings of the two square inner rings, and the two protein marking areas have the same shape but different sizes.

2. The integrated microwave-microfluidic chip according to claim 1, characterized in that: One end of the microfluidic channel is provided with a blood sample inlet, and the other end is provided with a blood sample outlet.

3. The integrated microwave-microfluidic chip according to claim 1, characterized in that: The forked microstrip line is connected to a data connection port, and the data connection port is used to connect to a host computer.

4. The method for preparing an integrated microwave-microfluidic chip according to any one of claims 1 to 3, characterized in that: The steps include: After setting a microwave sensor on a copper-based glass slide, the copper-based glass slide is cleaned and salted, and after marking the microwave sensor with an activated antibody solution, the antibody-coated microwave sensor is washed with deionized water and air-dried; After cleaning and salinizing the microfluidic chip, the activated antibody solution is labeled onto the microfluidic chip, and the antibody-coated microfluidic chip is washed with deionized water and air-dried; The microfluidic channel of the microfluidic chip and the protein labeling area on the microwave sensor are aligned and then bonded.

5. The method for preparing an integrated microwave-microfluidic chip according to claim 4, wherein: The method also includes welding a data connection port to the microwave sensor.

6. The method for preparing an integrated microwave-microfluidic chip according to claim 4, wherein: The manufacturing step of the microfluidic chip further includes: punching holes at both ends of the microfluidic channel of the microfluidic chip to obtain a blood sample inlet and a blood sample outlet.

7. A detection method using an integrated microwave-microfluidic chip according to any one of claims 1 to 3, characterized in that: The steps include: Transmitting a signal to a microwave sensor; The blood sample enters the microfluidic channel and flows through the protein labeling area. The specific antibodies in the protein labeling area capture the biomarkers in the blood. When the biomarker is captured by the specific antibody, it causes a signal change.

8. A detection method using an integrated microwave-microfluidic chip as claimed in claim 7, characterized in that: The signal change is linearly related to the amount of biomarker captured.

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