CN111351848B - Preparation method of sensor, sensor and detection method of sensor - Google Patents
Preparation method of sensor, sensor and detection method of sensor Download PDFInfo
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- CN111351848B CN111351848B CN202010196266.3A CN202010196266A CN111351848B CN 111351848 B CN111351848 B CN 111351848B CN 202010196266 A CN202010196266 A CN 202010196266A CN 111351848 B CN111351848 B CN 111351848B
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- G01N29/00—Investigating or analysing materials by the use of ultrasonic, sonic or infrasonic waves; Visualisation of the interior of objects by transmitting ultrasonic or sonic waves through the object
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention belongs to the technical field of instant examination, and particularly relates to a preparation method of a sensor for detecting early markers of cardiac injury, the sensor prepared by the preparation method and a detection method of the sensor. The sensor (only millimeter-scale due to small size of the piezoelectric thin-film resonator) prepared by the preparation method has small size, is beneficial to large-scale and low-cost manufacturing by adopting a semiconductor process, and can be integrated in wearable electronic equipment or other small-sized electronic equipment; in addition, the sensor prepared by the preparation method is used for detecting the early markers of the cardiac injury based on the quality sensitivity principle, so that the interference of the liquid sample is reduced, and the detection precision of the early markers of the cardiac injury is improved; in addition, the invention also provides a detection method of the sensor prepared based on the preparation method, which is beneficial to realizing the real-time measurement of the early markers of the cardiac injury.
Description
Technical Field
The invention belongs to the technical field of instant examination, and particularly relates to a preparation method of a sensor for detecting early markers of cardiac injury, the sensor prepared by the preparation method and a detection method of the sensor.
Background
Cardiovascular disease is one of the most serious diseases that endanger human health and life. Among them, acute myocardial infarction is the most common and dangerous. Early diagnosis and treatment of acute myocardial infarction is critical to reducing its mortality and improving patient prognosis. The early-stage cardiac injury marker is an important detection index for clinically diagnosing heart diseases such as myocardial infarction, myocardial ischemia, heart failure and the like.
The early-stage cardiac injury markers mainly comprise creatine kinase MB isozyme (CK-MB), cardiac troponin (cTn), heart-type fatty acid binding protein (h-FABP), B-type natriuretic peptide (BNP) and the like. Currently, most routine laboratory tests of early markers of cardiac injury are based on chemiluminescence, enzyme-linked immunosorbent assay (ELISA) and immunoturbidimetry. While these methods can provide accurate, reliable and quality controlled results, they require complex equipment, millilitre volumes of sample and specialized handling.
In recent years, with the rise of point-of-care testing (POCT) technology, the medical behavior has revolutionized. The POCT equipment has short analysis time and low sample consumption, and has important significance for diagnosing critical and important diseases such as acute myocardial infarction and the like. At present, a plurality of POCT early cardiac injury marker sensors promising for detecting cardiac biomarkers have been developed, including various principles such as electrochemistry, magnetism, fluorescence and the like. For example:
Patent document 3 discloses a method for preparing a label-free electrochemical sensor of cardiac troponin I and a method for detecting cTnI, wherein a novel label-free sensor is constructed by combining a substance having electrochemical activity with a biological immune reaction prepared on site, and the sensor is subjected to a specific reaction with a target molecule (cardiac marker cTnI) to obtain a cardiac marker antigen-antibody binding layer, thereby causing electrochemical activity disturbance and causing regular change of an output electrochemical signal.
However, the above technical solutions based on electrochemical, magnetic or fluorescent principles have the following disadvantages:
(1) the size of the device is large, and a micro integrated test system is difficult to form; (2) the test of the magnetic resistance or electrochemical detection principle is easily influenced by the dielectric property and the magnetic property of the liquid test sample, so that the final detection result is inaccurate.
Documents of the prior art
Patent document
Patent document 1: the publication number is: CN 108845146a, publication date: 11 and 20 months in 2018;
patent document 2: the publication number is: CN 108663525a, publication date: year 2018, month 10, day 16;
patent document 3: the publication number is: CN 110161100a, publication date: 23/08/2019;
patent document 4: the publication number is: CN 110044987a, publication date: year 2019, month 07, and day 23.
Disclosure of Invention
One of the objectives of the present invention is to provide a method for preparing a sensor for early marker detection of cardiac injury, so as to prepare a sensor capable of early marker detection of cardiac injury.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method of making a sensor for early marker detection of cardiac injury comprising the steps of:
I. preparing a piezoelectric film sensor;
depositing a silicon oxide layer on the surface of an upper electrode of the piezoelectric film sensor;
III, manufacturing a side wall of the micro-channel on the surface of the silicon oxide layer;
IV, arranging a glass cover plate on the upper surface of the side wall of the micro-channel to form the micro-channel;
and V, assembling the early cardiac injury marker antibody on the surface of the silicon oxide layer in the micro-flow channel.
Preferably, in step I, the piezoelectric thin film sensor includes a diaphragm type piezoelectric thin film sensor, a solid mount type piezoelectric thin film sensor having an acoustic reflection layer, or an air gap type structural piezoelectric thin film sensor.
Preferably, in step II, the silicon oxide layer is obtained by magnetron sputtering deposition.
Preferably, in the step III, the side wall of the micro flow channel is made of SU8 negative glue, polydimethylsiloxane or polyimide and is manufactured by adopting a common photoetching, soft photoetching or nano-imprinting method; the height of the side wall of the micro flow channel is 1-5 mm.
Preferably, in step IV, the glass cover plate and the surfaces of the side walls of the micro flow channels are treated with oxygen-containing particles before the glass cover plate is placed.
Preferably, in step V, the assembly process of the early cardiac injury marker antibody is as follows:
firstly, introducing deionized water and ethanol into a micro-channel to clean the surface;
then introducing an ethanol solution of aminopropyltriethoxysilane, and interacting with hydroxyl of the silicon oxide layer to form an amino surface;
further introducing glutaraldehyde aqueous solution for aldehyde group modification;
then introducing phosphate buffer solution of the early cardiac injury marker antibody to carry out covalent binding of the antibody;
and finally, introducing a phosphate buffer solution of bovine serum albumin to block the unbound aldehyde groups.
Preferably, in step V, the early marker antibody for cardiac injury assembled on the surface of the silica layer in the micro flow channel comprises creatine kinase MB isozyme, cardiac troponin, cardiac fatty acid binding protein or B-type natriuretic peptide antibody.
Another object of the present invention is to provide a sensor for detecting early markers of cardiac injury, which has a small size and can effectively improve the detection accuracy of the concentration of early markers of cardiac injury.
In order to achieve the purpose, the invention adopts the following technical scheme:
a sensor for detecting early markers of heart injury is prepared by the preparation method of the sensor.
It is a further object of the present invention to provide a method for detecting a sensor for early marker detection of cardiac injury, so as to realize real-time measurement of the concentration of early marker of cardiac injury.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method of detection of a sensor for early marker detection of cardiac damage, based on the above mentioned sensor;
the detection method of the sensor comprises the following steps:
s1. introducing pure serum sample into the micro flow channel, and measuring the resonance frequency of the piezoelectric film resonator;
taking the resonance frequency of the resonator in a stable state as a measured frequency baseline;
s2, sequentially introducing serum standard solutions containing the cardiac injury early markers with different concentrations into the micro-channel, and continuously measuring the resonant frequency of the piezoelectric thin-film resonator under the serum standard solutions of the cardiac injury early markers with different concentrations;
sequentially obtaining a plurality of change curves of the resonant frequency along with time through the processes;
s3. taking the stable value of the resonance frequency in each change curve, comparing with the frequency base line, and using the frequency shift value as the response of the sensor to obtain the concentration correction curve of the sensor to the early markers of cardiac injury;
wherein, the frequency shift value is a stable value of the resonance frequency as the frequency base line value;
s4. introducing the serum sample to be tested into the micro flow channel;
s5. continuously measuring the resonant frequency of the piezoelectric film resonator to obtain the variation curve of the resonant frequency with time, taking the frequency stable value on the variation curve, comparing with the frequency base line, and using the frequency shift value as the sensor response;
s6. the concentration of the marker corresponding to the early stage of cardiac injury in the sensor response is obtained against the concentration calibration curve in step s3.
Preferably, in step s2, after each passage of serum standard solutions of different concentrations of early cardiac injury markers, sodium dodecyl sulfate is used to recover the sensor after the adsorption of early cardiac injury marker antibodies.
The invention has the following advantages:
as described above, the present invention provides a method for manufacturing a sensor for early marker detection of cardiac injury, and the sensor manufactured by the method (only millimeter scale due to small size of the piezoelectric thin film resonator) has small size, is beneficial to large-scale and low-cost manufacturing by using a semiconductor process, and can be integrated in wearable electronic equipment or other small electronic equipment. The sensor prepared by the method is used for detecting the early markers of the cardiac injury based on the quality sensitivity principle, so that the interference of the liquid sample is reduced, and the detection precision of the early markers of the cardiac injury is improved; in addition, the invention also provides a detection method of the sensor based on the method, which is beneficial to realizing the real-time measurement of early markers of cardiac injury.
Drawings
FIG. 1 is a block flow diagram of a method for manufacturing a sensor for early marker detection of cardiac injury according to example 1 of the present invention;
FIG. 2 is a schematic structural view of a piezoelectric thin film sensor according to embodiment 1 of the present invention;
FIG. 3 is a schematic assembly diagram of a sensor for early marker detection of cardiac injury according to example 1 of the present invention;
FIG. 4 is a schematic structural diagram of a sensor for early marker detection of cardiac injury in example 2 of the present invention;
FIG. 5 is a block flow diagram of a detection method of a sensor for early marker detection of cardiac injury in example 3 of the present invention;
FIG. 6 is a frequency-time graph of a sensor for early marker detection of cardiac injury according to example 3 of the present invention against a standard solution of pure serum and serum of cardiac troponin (cTnI), an early marker of cardiac injury;
FIG. 7 is a graph showing the correction of the concentration of cardiac troponin (cTnI) in example 3 of the present invention.
FIG. 8 is a graph showing the results of comparison between the measurement performed by the detection method of example 3 of the present invention and the measurement performed by the conventional chemiluminescence method.
101-piezoelectric layer, 102-upper electrode, 103-lower electrode, 104-support layer, 105-silicon substrate, 106-silicon oxide layer, 107-micro flow channel side wall, 108-glass cover plate;
109-early cardiac injury marker antibody, 110-acoustic reflector, 111-air gap, 112-microchannel.
Detailed Description
The invention is described in further detail below with reference to the following figures and detailed description:
example 1
This example 1 describes a method for preparing a sensor for early marker detection of cardiac injury.
As shown in fig. 1, the preparation method of the sensor comprises the following steps:
I. and (4) preparing the piezoelectric film sensor.
The piezoelectric thin film sensor manufactured in this example 1 is, for example, a diaphragm type piezoelectric thin film sensor, as shown in fig. 2 (a).
The sensor adopts an aluminum nitride film as a piezoelectric layer 101, the c axis has an inclination angle of 24 degrees with the vertical direction, and the thickness is 1 micron. The upper electrode 102 is made of gold material, the lower electrode 103 is made of tungsten material, and the thickness of the upper electrode and the lower electrode is 100 nanometers.
The support layer 104 is a silicon nitride film with a thickness of 800 nm.
The silicon substrate 105 below the acoustic wave oscillation region is completely etched to form a diaphragm structure.
Of course, the piezoelectric thin film sensor manufactured in this embodiment 1 may also be a solid mount type piezoelectric thin film sensor having an acoustic reflection layer, as shown in fig. 2(b), or an air gap type structural piezoelectric thin film sensor, as shown in fig. 2 (c).
In fig. 2(b), reference numeral 110 denotes an acoustic reflection layer, and in fig. 2(c), reference numeral 111 denotes an air gap.
The following will specifically describe the manufacturing process of the sensor in this embodiment 1 by taking a diaphragm type piezoelectric thin film sensor as an example.
A silicon oxide layer 106 is deposited on the upper electrode surface of the manufactured piezoelectric thin film sensor (a diaphragm type piezoelectric thin film sensor is shown in fig. 3 (a)), as shown in fig. 3 (b).
The silicon oxide layer 106 is obtained by magnetron sputtering deposition, and the deposition thickness is 200 nm.
The function of the silicon oxide layer 106 is to provide a hydroxyl surface for assembly of sensitive antibodies while isolating the test liquid from the electrodes.
Fabricating micro flow channel sidewalls 107 on the surface of the silicon oxide layer 106, as shown in fig. 3 (c). As can be seen from FIG. 3(c), two micro flow channel side walls 107 are formed on the surface of the silicon oxide layer 106 in the present example 1.
The micro flow channel side wall 107 is preferably made of Polydimethylsiloxane (PDMS) by soft lithography using a common photoresist as a template. Of course, the micro flow channel side wall 107 may also be made of SU8 negative glue or Polyimide (PI) material.
The manufacturing process of the micro flow channel side wall 107 may also include, for example, a general photolithography or nanoimprint method.
The height of the microchannel side wall 107 is 1-5 mm, for example, 2 mm.
A glass cover plate 108 is provided on the upper surface of the micro flow channel side wall 107 to form the micro flow channel, as shown in fig. 3 (d).
Before the glass cover plate 108 is disposed, oxygen particles are used to treat the glass cover plate and the surface of the side wall of the micro flow channel, the treatment power density is 1 watt/square centimeter, the atmospheric pressure of oxygen is 20Pa, and the treatment time is 10 minutes.
The surface treatment of the oxygen-containing particles has the effects of ensuring the connection effect between the side wall 107 of the micro flow channel and the glass cover plate 108, ensuring the use reliability of the micro flow channel, and preventing the solution in the micro flow channel from leaking.
The present embodiment 1 has the following effects in designing a micro flow channel on a piezoelectric thin film sensor: the sensor prepared by the method can carry out real-time continuous measurement on the sample solution to be measured introduced into the micro-channel based on the quality sensitivity principle, is favorable for ensuring the accuracy of the measurement result, is favorable for replacing the solution and realizes the rapid measurement of the sample (without waiting for drying).
V. assemble early cardiac injury marker antibody 109 on the surface of the silicon oxide layer 106 in the microchannel, as shown in fig. 3 (e).
The assembly process of the early cardiac injury marker antibody is illustrated by taking cardiac troponin as an example, and specifically comprises the following steps:
the surface is first cleaned by passing deionized water and ethanol through the microchannel 112.
Then, an ethanol solution of Aminopropyltriethoxysilane (APTES) concentration of 2% was introduced and soaked for 60 minutes. An amino surface is formed due to the interaction of the silane groups of APTES with the hydroxyl groups of the silica layer.
And further introducing a 5% aqueous solution of glutaraldehyde for 30 minutes to modify aldehyde groups on the amino surface.
The covalent binding of the antibody was then carried out by introducing a solution of cardiac troponin (cTnI) in Phosphate Buffered Saline (PBS) at 10. mu.g/ml for 2 hours.
Finally, unbound aldehyde groups were blocked for 30 minutes by passing 0.1% Bovine Serum Albumin (BSA) in PBS to minimize non-specific binding effects. Through the above processes, the assembly process of the cardiac troponin antibody is realized.
The method for assembling creatine kinase MB isozyme (CK-MB), heart-type fatty acid binding protein (h-FABP), B-type natriuretic peptide (BNP) and other early cardiac injury marker antibodies is the same as the above method, and is not repeated here.
The sensor for detecting the early marker of cardiac injury can be prepared by the preparation method in the embodiment 1.
The sensor manufactured by the embodiment 1 has a small size, so that the sensor is beneficial to large-scale and low-cost manufacturing by adopting a semiconductor process and can be integrated in wearable electronic equipment or other small-sized electronic equipment.
In addition, the sensor prepared in this embodiment 1 performs the detection of the early markers of cardiac injury based on the quality sensitivity principle, which is beneficial to reducing the interference of the liquid sample itself, thereby improving the accuracy of the detection of the early markers of cardiac injury.
Example 2
This example 2 describes a sensor for early marker detection of cardiac injury, which is manufactured based on the method for manufacturing the sensor for early marker detection of cardiac injury of example 1.
As shown in fig. 4, the sensor includes a piezoelectric thin film sensor, a silicon oxide layer 106, a micro flow channel side wall 107, a glass cover plate 108, and an early cardiac injury marker antibody 109.
Wherein, the silicon oxide layer 106 is located on the surface of the upper electrode 102 of the piezoelectric thin film sensor.
The micro flow channel sidewall 107 is disposed on the surface of the silicon oxide layer 106.
The glass cover plate 108 covers the upper surface of the micro channel sidewall 107 and is connected to the micro channel sidewall 107 to form a micro channel 112.
The early cardiac injury marker antibody 109 is assembled on the surface of the silicon oxide layer 106 in the micro flow channel.
The early marker antibody 109 for cardiac injury in example 2 includes antibodies such as creatine kinase MB isozyme (CK-MB), cardiac troponin (cTnI), cardiac fatty acid binding protein (h-FABP), or B-type natriuretic peptide (BNP).
The advantages of the sensor of embodiment 2 have been described in more detail in embodiment 1, and are not described in detail here.
In this embodiment 2, different sensitive (i.e., early cardiac injury marker) antibodies can be respectively assembled for different sensors to form a sensor array, so that a plurality of early cardiac injury markers can be jointly detected at the same time.
Example 3
This example 3 describes a detection method of a sensor for early marker detection of cardiac injury, which is implemented based on the sensor for early marker detection of cardiac injury described in the above example 2.
The detection process of the early cardiac injury marker is described below by taking cardiac troponin as an example.
As shown in fig. 5, the detection method of the sensor for detecting the early-stage cardiac injury marker includes the following steps:
s1. pure serum sample is introduced into the micro flow channel 112 to measure the resonance frequency of the piezoelectric thin film resonator.
Wherein, the inlet and the outlet of the micro flow channel 112 respectively use the injection needle to input and output the liquid sample, and inject the liquid sample through the injection pump and the flow pump, and the resonator frequency is measured by using the network analyzer or the frequency measuring circuit.
Due to the mass and damping load, the resonator frequency of the thin film sensor is significantly reduced after the liquid (here, the above-mentioned pure serum sample) is introduced, and the resonance frequency in a steady state should be taken as a frequency baseline, as shown in fig. 6.
s2, sequentially introducing serum standard solutions containing different concentrations of cardiac troponin (cTnI) into the micro flow channel 112, and as the antibody and the antigen react, the resonance frequency of the resonator gradually decreases and stabilizes, as shown in fig. 6.
The resonance frequency of the resonator was measured continuously at different concentrations of serum standard solutions of early markers of cardiac injury.
Because a group of frequency change curves along with time can be obtained every time a serum standard solution of cardiac troponin (cTnI) with a certain concentration is introduced, a plurality of resonance frequency change curves along with time can be sequentially obtained through the process.
The serum standard solution concentration of cardiac troponin (cTnI) shown in FIG. 6 was 0.1 ng/ml.
After each time of introducing serum standard solution of cardiac troponin (cTnI) with different concentrations, 1% Sodium Dodecyl Sulfate (SDS) is needed to restore the sensor after the adsorption of the marker antibody in the early stage of heart injury, so that the sensor can be reused.
Of course, this embodiment 3 is not limited to the use of SDS, and other sensors that have the same or similar function to SDS and that are capable of recovering the adsorption of the marker antibody in the early stage of cardiac injury may be used, so that the sensor can be reused.
s3. the stable value of the resonance frequency in each variation curve is taken and compared with the frequency base line, and the frequency shift value is used as the response of the sensor to obtain the concentration correction curve of the sensor to the markers in the early stage of the heart injury, as shown in fig. 7.
Where the squares represent the data points obtained from actual testing (the lines above and below the squares represent the standard deviations of 5 measurements) and the dashed lines represent a linear fit to the test results. As can be seen from fig. 7, the concentration of cardiac troponin (cTnI) and the frequency shift value thereof are approximately linear in the logarithmic coordinate system. The frequency shift value is a stable value of the resonance frequency, which is a frequency base line value.
s4. the serum sample to be tested is introduced into the micro flow channel.
The serum sample to be tested in this example 3 is a human blood sample obtained by a standard blood collection procedure.
s5. the resonance frequency of the piezoelectric thin film resonator is measured continuously to obtain the variation curve of the resonance frequency with time, the frequency stability value on the variation curve is taken and compared with the frequency base line, and the frequency shift value is used as the sensor response.
s6. the concentration of the corresponding cardiac troponin (cTnI) in the sensor response is obtained against the concentration correction curve in step s3.
The above process is a method for detecting the concentration of cardiac troponin (cTnI).
The detection process of other early markers of cardiac injury, such as creatine kinase MB isozyme (CK-MB), heart-type fatty acid binding protein (h-FABP), B-type natriuretic peptide (BNP), is the same as the above method, and is not repeated here.
The embodiment 3 is beneficial to realizing real-time dynamic detection of early markers of cardiac injury, and the detection result is accurate and reliable, and the specific principle analysis is as follows: the liquid flowing in the test can wash away substances which are not bound to the antibody on the surface of the sensor, so that the nonspecific adsorption of the surface of the sensor is reduced, and the accuracy is improved. In addition, the micro-channel has fixed channel and cavity volume, thus the volume of the sample entering the sensitive area of the sensor can be accurately controlled, and the reliability and the repeatability of the test are improved.
The piezoelectric thin film resonator is manufactured by adopting a silicon semiconductor process and can be integrated in a miniature integrated test system, the dynamic measurement of the flowing liquid can realize continuous and automatic measurement with other components (such as blood separation, centrifugation and the like), and after one-time measurement is finished, new liquid (such as SDS) can be introduced to clean the surface, and multiple times of automatic repeated measurement can be realized.
In addition, in this example 3, the results of the detection by the method of the present invention were compared with the results of the conventional chemiluminescence method, and the comparison results are shown in FIG. 8. Wherein the squares represent the detection results of data points obtained by actual tests, and the dotted lines represent the results of a linear fit to the test results. The result detected by the method is compared with the result obtained by the conventional chemiluminescence method, and the result shows consistency, so that the detection method in the embodiment 3 of the invention has better detection accuracy.
It should be understood, however, that the description herein of specific embodiments is not intended to limit the invention to the particular forms disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. A method for detecting a sensor for early marker detection of cardiac injury, based on a sensor for early marker detection of cardiac injury, the method for preparing the sensor comprising the steps of:
I. preparing a piezoelectric film sensor;
depositing a silicon oxide layer on the surface of an upper electrode of the piezoelectric film sensor;
III, manufacturing a micro-channel side wall on the surface of the silicon oxide layer;
IV, arranging a glass cover plate on the upper surface of the side wall of the micro-channel to form the micro-channel;
v, assembling early cardiac injury marker antibodies on the surface of the silicon oxide layer in the micro-channel; it is characterized in that the preparation method is characterized in that,
the detection method of the sensor comprises the following steps:
s1, introducing a pure serum sample into the micro-channel, and measuring the resonant frequency of the piezoelectric thin-film resonator;
taking the resonance frequency of the resonator in a stable state as a measured frequency baseline;
s2, sequentially introducing serum standard solutions containing the cardiac injury early markers with different concentrations into the micro-channel, and continuously measuring the resonant frequency of the piezoelectric thin-film resonator under the serum standard solutions of the cardiac injury early markers with different concentrations;
sequentially obtaining a plurality of change curves of the resonant frequency along with time through the processes;
s3, taking the stable value of the resonance frequency in each change curve, then comparing with the frequency base line, and using the frequency shift value as the response of the sensor to obtain the concentration correction curve of the sensor to the early markers of the heart injury;
wherein frequency shift value = frequency baseline value — stable value of resonant frequency;
s4, introducing a serum sample to be detected into the micro-channel;
s5, continuously measuring the resonant frequency of the piezoelectric film resonator to obtain the change curve of the resonant frequency along with time, taking the frequency stable value on the change curve, comparing with the frequency base line, and using the frequency movement value as the sensor response;
s6 the sensor response is obtained in response to the early marker concentration of cardiac injury, against the concentration calibration curve in step s3.
2. The detection method of a sensor according to claim 1,
in the step I, the piezoelectric film sensor includes a diaphragm type piezoelectric film sensor, a solid assembly type piezoelectric film sensor having an acoustic reflection layer, or an air gap type structural piezoelectric film sensor.
3. The detection method of a sensor according to claim 1,
in the step II, the silicon oxide layer is obtained by adopting a magnetron sputtering deposition method.
4. The detection method of a sensor according to claim 1,
in the step III, the side wall of the micro-channel is made of SU8 negative glue, polydimethylsiloxane or polyimide and is manufactured by common photoetching and soft photoetching; the height of the side wall of the micro flow channel is 1-5 mm.
5. The detection method of a sensor according to claim 1,
in step IV, the glass cover plate and the surface of the side wall of the micro flow channel are treated with oxygen-containing particles before the glass cover plate is placed.
6. The detection method of a sensor according to claim 1,
in step V, the assembly process of the early heart injury marker antibody is as follows:
firstly, introducing deionized water and ethanol into the micro-channel to clean the surface;
then introducing an ethanol solution of aminopropyltriethoxysilane, and interacting with hydroxyl of the silicon oxide layer to form an amino surface;
further introducing glutaraldehyde aqueous solution for aldehyde group modification;
then introducing phosphate buffer solution of the early cardiac injury marker antibody to carry out covalent binding of the antibody;
and finally, introducing a phosphate buffer solution of bovine serum albumin to block the unbound aldehyde groups.
7. The detection method of a sensor according to claim 1,
in the step V, the early cardiac injury marker antibody assembled on the surface of the silicon oxide layer in the micro-channel comprises creatine kinase MB isozyme, cardiac troponin, cardiac fatty acid binding protein or B-type natriuretic peptide antibody.
8. The detection method of a sensor according to claim 1,
in step s2, after each time of introducing the serum standard solution of the early cardiac injury marker with different concentrations, sodium dodecyl sulfate is used to recover the sensor after the adsorption of the early cardiac injury marker antibody.
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| CN103472129B (en) * | 2013-09-17 | 2017-01-11 | 天津大学 | Resonance sensor for fluid environment detection |
| US9910015B2 (en) * | 2014-04-14 | 2018-03-06 | Texas Instruments Incorporated | Sensor array chip with piezoelectric transducer including inkjet forming method |
| EP3365669B8 (en) * | 2015-10-21 | 2024-03-13 | Qorvo Us, Inc. | Resonator structure with enhanced reflection of shear and longitudinal modes of acoustic vibrations |
| US10352904B2 (en) * | 2015-10-26 | 2019-07-16 | Qorvo Us, Inc. | Acoustic resonator devices and methods providing patterned functionalization areas |
| JP6882280B2 (en) * | 2015-11-06 | 2021-06-02 | コーボ ユーエス，インコーポレイティド | Acoustic resonator devices, as well as manufacturing methods that provide airtightness and surface functionalization. |
| WO2017156127A1 (en) * | 2016-03-11 | 2017-09-14 | Qorvo Us, Inc. | Baw sensor fluidic device with increased dynamic measurement range |
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| CN109012771B (en) * | 2018-07-23 | 2020-06-09 | 武汉大学 | Full-transparent microfluidic acoustic bulk wave chip and preparation method thereof |
| CN110061715B (en) * | 2019-03-29 | 2020-07-07 | 山东科技大学 | Method for manufacturing piezoelectric thin film resonator on non-silicon substrate |
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2020
- 2020-03-19 CN CN202010196266.3A patent/CN111351848B/en active Active
- 2020-04-27 WO PCT/CN2020/087123 patent/WO2021184494A1/en active Application Filing
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| CN111351848A (en) | 2020-06-30 |
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