CN109187974A - Carcinomebryonic antigen sensor and preparation method thereof, carcinomebryonic antigen concentration detection method - Google Patents
Carcinomebryonic antigen sensor and preparation method thereof, carcinomebryonic antigen concentration detection method Download PDFInfo
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Abstract
本发明公开了一种癌胚抗原传感器，在压电材料上制作输入换能器和输出换能器构建声表面波传感器，并利用沉积在输入换能器和输出换能器之间的金膜来固定癌胚抗体，通过癌胚抗体实现癌胚抗原浓度检测。声表面波传感器的制作成本低廉，并结合癌胚抗体与抗原的特异性，实现癌胚抗原浓度的高精度检测。本发明的癌胚抗原传感器的制作方法，通过在压电材料层上制作输入换能器和输出换能器；再在输入换能器和输出换能器之间的压电材料上沉积金膜，最后在金膜上固定癌胚抗体；完成癌胚抗原传感器的制作。本发明的癌胚抗原浓度检测方法，利用癌胚抗原传感器实现癌胚抗原浓度检测，根据第一频率和第二频率之差与癌胚抗原浓度的常数对数之间的线性关系获取待测液体的癌胚抗原浓度。
The invention discloses a carcinoembryonic antigen sensor. An input transducer and an output transducer are fabricated on piezoelectric materials to construct a surface acoustic wave sensor, and a gold film deposited between the input transducer and the output transducer is used. To fix the carcinoembryonic antibody, the concentration of carcinoembryonic antigen can be detected by the carcinoembryonic antibody. The fabrication cost of the surface acoustic wave sensor is low, and combined with the specificity of the carcinoembryonic antibody and the antigen, the high-precision detection of the carcinoembryonic antigen concentration can be realized. The manufacturing method of the carcinoembryonic antigen sensor of the present invention comprises the steps of making an input transducer and an output transducer on a piezoelectric material layer; and then depositing a gold film on the piezoelectric material between the input transducer and the output transducer , and finally immobilize the carcinoembryonic antibody on the gold film; complete the fabrication of the carcinoembryonic antigen sensor. The carcinoembryonic antigen concentration detection method of the present invention utilizes the carcinoembryonic antigen sensor to realize the carcinoembryonic antigen concentration detection, and obtains the liquid to be tested according to the linear relationship between the difference between the first frequency and the second frequency and the constant logarithm of the carcinoembryonic antigen concentration carcinoembryonic antigen concentration.
Description
Technical field
The present invention relates to sensor field, especially a kind of carcinomebryonic antigen sensor and preparation method thereof, carcinomebryonic antigen are dense
Spend detection method.
Background technique
Surface acoustic wave sensor is a kind of MEMS device, has low cost, higher sensitivity, real-time detection, Breakup of Liquid Ring
The advantages that border is detected.The surface of surface acoustic wave sensor can generate the vibration of high frequency, micro-nano level amplitude, when big in liquid
Molecule or particle can generate interference to the vibration of above-mentioned high frequency when being deposited on device surface, by measuring this interference, finally
Learn the content of macromolecular or particle in a liquid.
Carcinomebryonic antigen is a broad spectrum activity tumor markers, it can reflect the presence of kinds of tumors to people.Work as patient
When having the cancers such as colorectal cancer, breast cancer and lung cancer, carcinomebryonic antigen can be significantly raised in the intracorporal content of people.Therefore, it detects this
Tumor markers can early diagnosis to patient, curative effect judgement, progression of the disease, monitoring and prognosis estimation extend efficient help.
But the testing liquid very low for carcinomebryonic antigen content, existing detection device more difficult low cost, accurately completion detection,
The sensitivity of existing detection device can not be supported to detect.
Summary of the invention
The present invention is directed to solve at least some of the technical problems in related technologies.For this purpose, of the invention
One purpose is to provide a kind of highly sensitive, at low cost carcinomebryonic antigen sensor and preparation method thereof, the inspection of carcinomebryonic antigen concentration
Survey method.
The technical scheme adopted by the invention is that: a kind of carcinomebryonic antigen sensor, including substrate, the carcinomebryonic antigen sensing
Device further includes setting piezoelectric material layer over the substrate, the input energy converter being arranged on the surface of the piezoelectric material layer
And output transducer, be arranged in it is described input energy converter and the output transducer between golden film and be fixed on the gold
Cancer embryo antibody on film, the input energy converter are used to the rf signal of input being excited into surface acoustic wave, and the output is changed
Energy device is for being converted to rf signal for received surface acoustic wave and exporting.
Further, the input energy converter and/or output transducer are interdigital transducer.
Further, the interdigital transducer has 20-80200 to interdigital electrode pair.
Further, the interdigital width range of the interdigital transducer and finger spacing range are 1-50 microns.
Further, the substrate and/or piezoelectric material layer are piezoelectric monocrystal or piezoelectric membrane.
Further, the thickness range of the golden film is 100nm-200nm.
It is of the present invention another solution is that a kind of production method of carcinomebryonic antigen sensor, be applied to described
A kind of carcinomebryonic antigen sensor, comprising the following steps:
Production input energy converter and output transducer on piezoelectric material layer；
Golden film is made on the piezoelectric material layer, the golden film is arranged in the input energy converter and the output transducing
Between device；
Cancer embryo antibody is fixed in the golden film.
Further, the golden film is made on the piezoelectric material layer using physical gas phase deposition technology.
Further, the method that cancer embryo antibody is fixed in the golden film includes:
On the surface of the golden film, addition hydrosulphonyl silane alcoholic solution is reacted；
On the surface of the golden film, addition cysteamine ethanol solution is reacted；
On the surface of the golden film, addition cancer embryo antibody-solutions are reacted to fix cancer embryo antibody.
It is of the present invention another solution is that a kind of cancer embryo of carcinomebryonic antigen sensor described in a kind of application is anti-
Original content detection method, comprising the following steps:
The centre frequency of carcinomebryonic antigen sensor when being not added with testing liquid is set into first frequency；
Testing liquid is added in the golden film for being fixed with cancer embryo antibody to be reacted；
After fully reacting, the centre frequency for obtaining the carcinomebryonic antigen sensor is second frequency.
According between the difference of the frequency of the first frequency and the second frequency and the constant logarithm of carcinomebryonic antigen concentration
Linear relationship obtain the carcinomebryonic antigen concentration of the testing liquid.
The beneficial effects of the present invention are:
Carcinomebryonic antigen sensor of the invention, setting input energy converter and output transducer construct surface acoustic wave sensor,
The cost of manufacture of surface acoustic wave sensor is cheap, and combines the specificity of cancer embryo antibody and antigen, realizes carcinomebryonic antigen concentration
High-precision detects.The present invention overcomes the prior art to cannot achieve the defect of low concentration carcinomebryonic antigen Concentration Testing, and it is anti-to improve cancer embryo
The sensitivity of original content detection and Monitoring lower-cut.
The production method of carcinomebryonic antigen sensor of the invention, by making input energy converter and defeated on piezoelectric material layer
Energy converter constructs surface acoustic wave sensor out；Make golden film on piezoelectric material layer again, golden film setting is in input energy converter and defeated
Out between energy converter；Cancer embryo antibody is finally fixed in golden film；Complete the production of carcinomebryonic antigen sensor.
Carcinomebryonic antigen concentration detection method of the invention realizes carcinomebryonic antigen Concentration Testing using carcinomebryonic antigen sensor,
The centre frequency of carcinomebryonic antigen sensor when being not added with testing liquid is set as first frequency, is being fixed with cancer embryo antibody
Testing liquid to be added in golden film to be reacted, after fully reacting, the centre frequency for obtaining carcinomebryonic antigen sensor is second frequency,
Institute is obtained using the linear relationship between the difference of the frequency of first frequency and second frequency and the constant logarithm of carcinomebryonic antigen concentration
State the carcinomebryonic antigen concentration of testing liquid.
Detailed description of the invention
Specific embodiments of the present invention will be further explained with reference to the accompanying drawing:
Fig. 1 is an a kind of specific embodiment stereoscopic schematic diagram of carcinomebryonic antigen sensor in the present invention；
Fig. 2 is the reaction of cancer embryo antibody and carcinomebryonic antigen fixed in a kind of golden film of carcinomebryonic antigen sensor in the present invention
Process schematic；
Fig. 3 be according to the present invention in a kind of structural schematic diagram of one embodiment of micro-fluidic chip；
Fig. 4 be according to the present invention in a kind of structural schematic diagram of one embodiment of micro fluidic device；
Fig. 5 is the perspective view of Fig. 4；
Fig. 6 be according to the present invention in a kind of structural schematic diagram of another embodiment of micro fluidic device；
Fig. 7 be according to the present invention in a kind of structural schematic diagram of another embodiment of micro fluidic device；
Fig. 8 is the side view of Fig. 7 embodiment；
Fig. 9 be according to the present invention in a kind of structural schematic diagram of another embodiment of micro fluidic device；
Figure 10 be according to the present invention in a kind of structural schematic diagram of another embodiment of micro fluidic device；
Figure 11 is that the difference of corresponding first frequency and second frequency is at any time when adding the carcinomebryonic antigen solution of various concentration
The relational graph of variation；
Figure 12 is that first frequency and the difference of second frequency and one specific embodiment of constant logarithm of carcinomebryonic antigen concentration are linear
Relational graph.
Specific embodiment
It should be noted that in the absence of conflict, the features in the embodiments and the embodiments of the present application can phase
Mutually combination.
A kind of carcinomebryonic antigen sensor, including substrate, carcinomebryonic antigen sensor further include the piezoresistive material of setting on substrate
The bed of material, the input energy converter being arranged on the surface of piezoelectric material layer and output transducer, setting are in input energy converter and output
Golden film between energy converter and the cancer embryo antibody being fixed in golden film, the rf signal that input energy converter is used to input
It is excited into surface acoustic wave, output transducer is used to being converted to received surface acoustic wave into rf signal output.Wherein, input is changed
Energy device and output transducer and piezoelectric material layer constitute surface acoustic wave sensor, due to the mass loading effect energy of sensor surface
Moment and significant the output signal for influencing surface acoustic wave sensor, using surface acoustic wave sensor to the characteristic of surface-sensitive,
Carcinomebryonic antigen is adsorbed by the cancer embryo antibody specificity being bundled on golden film surface, the disturbed degree of sensor surface is visited
It surveys, to learn the concentration of contained carcinomebryonic antigen in liquid, the prior art is overcome to cannot achieve the inspection of low concentration carcinomebryonic antigen concentration
The defect of survey improves the sensitivity and detection of carcinomebryonic antigen Concentration Testing with the concentration of highly sensitive real-time detection carcinomebryonic antigen
Lower limit.In addition, since surface acoustic wave sensor can be mass produced using mature MEMS technology, carcinomebryonic antigen
The cost of manufacture of sensor is cheap.
As the further improvement of technical solution, in the present embodiment, substrate and/or piezoelectric material layer be piezoelectric monocrystal (such as
Quartz, lithium tantalate etc.) or piezoelectric membrane (such as ZnO, AlN).In addition, the thickness range of golden film is 100nm-200nm.Input is changed
Energy device and/or output transducer are interdigital transducer.Interdigital transducer has 20-200 to interdigital electrode pair.Interdigital transducer
Interdigital width range and finger spacing range are 1-50 microns.Interdigital transducer can be made of metallic films such as Al and Au.
When not inputting testing liquid, the centre frequency of carcinomebryonic antigen sensor is first frequency, it by piezoelectric material layer material with
The design of interdigital transducer determines that for the same device, it is a fixed value.
It is one of a kind of carcinomebryonic antigen sensor in the present invention with reference to Fig. 1, Fig. 1 as the further improvement of technical solution
Specific embodiment stereoscopic schematic diagram；In the present embodiment, lithium tantalate is cut as piezoelectric material layer 1 using 36 ° of-Y, inputs energy converter 2
It is arranged on piezoelectric material layer 1 with output transducer 3, the setting of golden film 4 is among input energy converter 2 and output transducer 3, input
Energy converter 2 and output transducer 3 are interdigital transducer, cancer embryo antibody 5 are fixed in golden film 4, cancer embryo antibody 5 is for specificity
Adsorb carcinomebryonic antigen 6.
Then, the present invention also provides a kind of production methods of carcinomebryonic antigen sensor, anti-applied to a kind of above-mentioned cancer embryo
Original sensor, comprising the following steps: production input energy converter and output transducer on piezoelectric material layer；On piezoelectric material layer
Golden film is made, golden film setting is between input energy converter and output transducer；Cancer embryo antibody is fixed in golden film.
With reference to Fig. 1 and Fig. 2, Fig. 2 be cancer embryo antibody fixed in a kind of golden film of carcinomebryonic antigen sensor in the present invention and
The reaction process schematic diagram of carcinomebryonic antigen；The production method that carcinomebryonic antigen sensor is specifically described below:
Piezoelectric material layer 1 of the lithium tantalate as carcinomebryonic antigen sensor is cut using 36 ° of mature-Y, by piezoelectric material
Layer 1 on depositing Al film and using photoetching technique prepare interdigital electrode, that is, interdigital transducer, i.e., input energy converter 2 and output transducing
Device 3, the interdigital width and finger spacing of interdigital electrode are 5 microns, aperture 4mm, and the finger number of two groups of interdigital electrodes is 80
It is right.
Magnetron sputtering technique is recycled to sputter 100- in the surface-sensitive areas (10mm × 8mm) of carcinomebryonic antigen sensor
The golden film 4 of 200nm or so, the setting of golden film 4 is among input energy converter and output transducer.
The NaOH solution for the use of 10ml concentration being successively 1M, the HCl solution that 10ml concentration is 1M, the piranha of 10ml are molten
Liquid cleans the golden film surface of carcinomebryonic antigen sensor, and duration of the reaction is respectively 20 minutes, 5 minutes, 5 minutes, finally with super
The entire carcinomebryonic antigen sensor of pure water rinsing.
Carcinomebryonic antigen sensor surface is cleaned using plasma degumming machine.It is 300W, working time that power, which occurs, for plasma
For 2 minutes, O2 flow be 30sccm.
Take the silane ethanol solution drop that 1ml concentration is 2mM on the golden film surface of carcinomebryonic antigen sensor, normal temperature and pressure is stood
18-24h.Wherein, the reaction equation being related to are as follows:
2Si(OH)3SH+Au→Si(OH)3SAu+H2↑。
The surface of carcinomebryonic antigen sensor is cleaned with 50ml deionized water and is 2mM's with 2ml concentration after being dried with nitrogen, is taken
For cysteamine ethanol solution drop on the golden film surface of carcinomebryonic antigen sensor, normal temperature and pressure stands reaction 18-24h.The reaction being related to
Equation are as follows:
3C2H7NS+Si(OH)3SAu→C2H4NS-Si-SAu+H2O。
Take the cancer embryo antibody-solutions (the i.e. Anti-CEA of cancer embryo antibody) that 5 μ l concentration are 200 μ g/ml with 2ml pH=7.4's
It is added dropwise with test tube in the golden film surface of carcinomebryonic antigen sensor, is stood after PBS buffer (bioactivity for keeping antibody) mixing
React 36-48h.The reaction equation being related to are as follows:
Successively using the surface of the PBS buffer solution of 10ml and the ultrapure water carcinomebryonic antigen sensor of 10ml, nitrogen is used
After drying, the BSA solution that concentration is 1M is added dropwise in carcinomebryonic antigen sensor surface, is reacted 2 hours under normal temperature and pressure.So far, exist
Cancer embryo antibody is secured in golden film, completes the production of carcinomebryonic antigen sensor.
Finally, the present invention also provides a kind of carcinomebryonic antigen concentration detection method using above-mentioned carcinomebryonic antigen sensor, packet
Include following steps:
The centre frequency of carcinomebryonic antigen sensor when being not added with testing liquid is set as first frequency；
Testing liquid is added in the golden film for being fixed with cancer embryo antibody to be reacted；
After fully reacting, the centre frequency for obtaining carcinomebryonic antigen sensor is second frequency.
According to the linear pass between the difference of the frequency of first frequency and second frequency and the constant logarithm of carcinomebryonic antigen concentration
System obtains the carcinomebryonic antigen concentration of the testing liquid.
Specifically, it with reference to Fig. 1 and Fig. 2, in the present embodiment, inputs energy converter and output transducer is interdigital transducer,
When on carcinomebryonic antigen sensor without addition testing liquid, the centre frequency of carcinomebryonic antigen sensor is first frequency.When certain
When the radiofrequency signal of frequency is applied on input energy converter, due to the inverse piezoelectric effect of piezoelectric material layer, the table of piezoelectric material layer
Face can be inspired surface acoustic wave and transmit along the surface of piezoelectric material layer, by the sensitivity of cancer embryo antibody and golden film composition
Film, when the carcinomebryonic antigen (CEA) in testing liquid is exempted from by the cancer embryo antibody (Anti-CEA) that golden film surface is fixed by specificity
After epidemic disease absorption, amplitude and the frequency of surface acoustic wave can change because of the change of surface quality, therefore carcinomebryonic antigen sensor is in cancer
After embryo antibody and carcinomebryonic antigen fully reacting, the centre frequency (second frequency) of carcinomebryonic antigen sensor can reduce, according to first
Relationship existing for frequency and the difference of second frequency and carcinomebryonic antigen concentration, it is final it can be seen that in liquid carcinomebryonic antigen concentration.Tool
Body, it can be to the carcinomebryonic antigen solution of various concentration known to the same carcinomebryonic antigen biosensor assay multiple groups, due to same
The first frequency of carcinomebryonic antigen sensor fix and it is known that addition various concentration carcinomebryonic antigen solution when golden film surface,
Multiple second frequencies of the carcinomebryonic antigen solution of available corresponding various concentration, according to multiple second frequencies and first frequency
Multiple difference on the frequencies and known carcinomebryonic antigen concentration, the first frequency of the available carcinomebryonic antigen sensor and second frequency
Linear relationship between the difference of frequency and the constant logarithm of carcinomebryonic antigen concentration.
The linear relationship test between above-mentioned acquisition difference on the frequency and the constant logarithm of carcinomebryonic antigen concentration is specifically described below
Process:
In actual use, carcinomebryonic antigen sensor is arranged on micro-fluidic chip, is root with reference to Fig. 3, Fig. 3 specifically
According to the structural schematic diagram of one embodiment of micro-fluidic chip a kind of in the present invention；Micro-fluidic chip it include signal source, first
Load circuit, the first connecting element, two or more miniflow cavity (not shown), carcinomebryonic antigen sensor, the second connection member
Part and the second load circuit, wherein a part of miniflow cavity is input miniflow cavity, for injecting testing liquid in cancer embryo
The detection zone of antigen sensor, detection zone are the region where golden film；Another part of miniflow cavity is output miniflow cavity, is used
When the liquid that has detected of emptying, detection, signal source, the first load circuit, the first connecting element, carcinomebryonic antigen sensor, the
Two connecting elements and the second load circuit are sequentially connected in series to form closed circuit, input electricity of the signal source in carcinomebryonic antigen sensor
Road, which inputs on energy converter, applies rf signal, and passes through the output circuit of carcinomebryonic antigen sensor, that is, output transducer output
Rf signal.In the present embodiment, signal source is Network Analyzer, for exporting rf signal, and with input energy converter and
Output transducer is all connected with to obtain the centre frequency of carcinomebryonic antigen sensor.
As the further improvement of micro-fluidic chip, the carcinomebryonic antigen sensor of micro-fluidic chip include input circuit (i.e.
Input energy converter) and output circuit (i.e. output transducer), while the first connecting element may include that the first probe and second are visited
Needle, is connected to the both ends of the input circuit of carcinomebryonic antigen sensor, the second connecting element may include third probe and
4th probe is connected to the both ends of the output circuit of carcinomebryonic antigen sensor.Wherein the first probe, the second probe,
Three probes and the 4th probe can be spring probe.Wherein, the first load circuit and the second load circuit can pass through sub-miniature A connector
The both ends of signal source are connected to, so that micro-fluidic chip can be adapted for the microwave regime of frequency range direct current to 26.5GHz
Using, therefore bandwidth, performance are excellent, highly reliable, the service life is long.In addition, micro-fluidic chip can also be pumped including miniflow and syringe,
Miniflow pump injects testing liquid to miniflow cavity to inject micro testing liquid by syringe when wherein detecting, and detection is completed
Miniflow pump injects PBS buffer solution to miniflow cavity to which emptying has detected liquid by syringe afterwards.
Further, as shown in Figure 4 and Figure 5, Fig. 4 be according to the present invention in a kind of one embodiment of micro fluidic device
Structural schematic diagram；Fig. 5 is the perspective view of Fig. 4；A kind of micro fluidic device comprising the micro-fluidic chip, and it is described micro-
Flow control apparatus can also include top plate 100, middle plate 200 and pedestal 300, and wherein top plate 100 is provided with for the first connection
The first through hole of element and the insertion of the second connecting element (not shown), such as 101,102,103 and 104, middle plate 200 is provided with
Two or more miniflow cavities, such as 201 and 202, and supply the second of the first connecting element and the insertion of the second connecting element
Through-hole, such as 203,204,205 and 206 is provided with carcinomebryonic antigen sensor (not shown) on pedestal 300.Top plate 100 is also set
It is equipped with the third through-hole for injecting and emptying for testing liquid, such as 105 and 106, the quantity of third through-hole and the number of miniflow cavity
It measures identical.When detection, top plate 100, middle plate 200 are directed at 300 with pedestal, so that a part and input of third through-hole
The starting point alignment of miniflow cavity is to inject another part of testing liquid and third through-hole and the end of output miniflow cavity
Alignment is to empty the liquid detected, for example, the position of through-hole 105 corresponds to the starting point of input miniflow cavity 201 and leads to
The position in hole 106 corresponds to the end of output miniflow cavity 202.When detection, the first connecting element and the second connecting element are worn
Cross first through hole, the second through-hole is connected to the both ends of carcinomebryonic antigen sensor, for example, the first connecting element can pass through it is upper
The through-hole 101 and 102 of laminate and pass through middle plate through-hole 203 and 204 and be connected to one end of carcinomebryonic antigen sensor i.e.
Energy converter is inputted, the second connecting element can pass through the through-hole 103 and 104 of top plate and pass through 205 He of through-hole of middle plate
206 and be connected to the other end i.e. output transducer of carcinomebryonic antigen sensor.Wherein, although Fig. 4 is merely illustrated with fixed number
The case where through-hole and miniflow cavity of amount, but micro fluidic device of the invention at the middle and upper levels plate, on middle plate the quantity of through-hole and
The quantity of miniflow cavity can specifically be set according to practical application.Wherein the middle plate 200 of micro fluidic device corresponds to cancer embryo
Hollow reaction zone 207 can be set in the detection zone of antigen sensor, that is, golden film region position.Preferably, reaction zone
Size can be equal or slightly larger than carcinomebryonic antigen sensor detection zone size.As shown in fig. 6, Fig. 6 is according to the present invention
A kind of structural schematic diagram of another embodiment of middle micro fluidic device；Wherein carcinomebryonic antigen sensor 400 includes input circuit
401 and output circuit 402, preferably the first connecting element may include the first probe and the second probe, the second connecting element packet
Third probe and the 4th probe (not shown) are included, the first probe and the second probe are connected to carcinomebryonic antigen sensing when detection
The both ends of the input circuit 402 of device, i.e. two input terminals of the both ends of input energy converter namely interdigital electrode, can be used for inputting
Rf signal；Third probe and the 4th probe are connected to the both ends of the output circuit 402 of carcinomebryonic antigen sensor, i.e., defeated
Two output ends of the both ends of energy converter namely interdigital electrode out, can be used for exporting rf signal.Wherein the first probe,
Two probes, third probe and the 4th probe can be spring probe.The micro fluidic device can also include that miniflow is pumped and injected
Device is wherein directed at the third through-hole for inputting miniflow cavity in the output end of syringe and top plate, such as the sealing of through-hole 105 connects
It connects, so that miniflow pump is by syringe injection testing liquid to miniflow cavity to inject micro testing liquid when detection.
Further, as shown in fig. 7, Fig. 7 be according to the present invention in a kind of knot of another embodiment of micro fluidic device
Structure schematic diagram；Wherein the micro fluidic device can also include sealing ring 500, and the setting of sealing ring 500 is passed in carcinomebryonic antigen
On sensor 400 and surround the detection zone 403 of carcinomebryonic antigen sensor 400.Preferably, the size of sealing ring 500 is equal to or bigger
In the size of the detection zone 403 of carcinomebryonic antigen sensor 400.Preferably, the silica gel that sealing ring 500 can be stable for chemical property
Rubber ring.Wherein first through hole is preferably four on top plate 100, such as 101,102,103 and 104, and middle plate 200
Upper second through-hole is preferably four, such as 203,204,205 and 206, so that the first probe 601, the second probe 602,
Three probes 603 and the 4th probe 604 can pass through top plate 100 and middle plate 200 to be connected to carcinomebryonic antigen sensor 400
Both ends.For example, the first probe 601 and the second probe 602 are connected to the both ends of the input circuit of carcinomebryonic antigen sensor 400, together
When the second probe 603 and the 4th probe 604 be connected to carcinomebryonic antigen sensor 400 output circuit both ends.As shown in Figure 8
(to put it more simply, part-structure is not shown), Fig. 8 are the side views of Fig. 7 embodiment；Pedestal 300 may be provided with groove 301 and 302,
Carcinomebryonic antigen sensor 400 to be directly or indirectly fixed on pedestal 300, which can be the mode of buckle, with
So that carcinomebryonic antigen sensor 400 is so detachable that be fixed on pedestal 300, therefore micro fluidic device is reusable.Such as Fig. 9 institute
Show (to put it more simply, part-structure is not shown), Fig. 9 be according to the present invention in a kind of knot of another embodiment of micro fluidic device
Structure schematic diagram；First load circuit and the second load circuit can be connected to signal by two sub-miniature A connectors 701 and 702
The both ends in source.The micro fluidic device of the present embodiment when detecting can be by magnetic attracting device by the top plate of micro fluidic device, middle layer
Plate and pedestal are closed together.(to put it more simply, part-structure is not shown) as shown in Figure 10, Figure 10 are middle a kind of according to the present invention
The structural schematic diagram of another embodiment of micro fluidic device；Wherein top plate 100 may be provided with magnet placing groove, such as 107,
108,109 and 110, while may also set up corresponding magnet placing groove on pedestal, such as 303,304,305 and 306.Magnet is put
It sets and may be provided with magnetic grain in slot.Wherein middle plate 200 can be combined together with top plate 100, it is preferable that can be tied by heat
It is legal to be combined together middle plate 200 and top plate 100.In this way, micro fluidic device has a semi-enclosed reaction zone
207.Protrusion, such as 307,308,309 and 310 have can be set in its insole board 300, while middle plate 200 can accordingly be set
It is equipped with through-hole, such as 208,209,210 and 211.When micro fluidic device closure, the protrusion on pedestal 300 can penetrate middle layer
Through-hole on plate 200 is middle plate 200 to be aligned with the corresponding region of pedestal 300, while the magnet in top plate 100 can be with
Magnet in pedestal 300 is attracting to compress entire micro fluidic device.Although Figure 10 show the fixed magnet placing groove of quantity with
Protrusion, but magnet placing groove of the invention can be arranged according to the actual situation and arbitrarily with the quantity of protrusion and position.Wherein
Micro fluidic device can be made by the stable acrylic of, chemical property preferable using biocompatibility.
So far, it with reference to Fig. 2, Fig. 3 and Fig. 4, is tried using the carcinomebryonic antigen solution that micro fluidic device can carry out various concentration
Test, firstly, carcinomebryonic antigen sensor after ultrapure water washes down surface with being dried with nitrogen, be put into micro-fluidic chip；Then by PBS
Buffer injects the detection zone of carcinomebryonic antigen sensor, can remove the impurity and protein on detection zone surface, observes the second frequency
The rising of rate；And over time, second frequency varies less.
With PBS solution configured various concentration carcinomebryonic antigen solution (in the present embodiment, the concentration of carcinomebryonic antigen solution point
It Wei 0g/ml, 0.5ng/ml, 1ng/ml, 5ng/ml, 10ng/ml).
Above-mentioned solution is passed sequentially through in micro-fluidic chip injection detection zone, every time with 1ml/ minutes speed injection 2ml
Solution.The ingredient of solution is PBS buffer solution and carcinomebryonic antigen at this time.It is solid with chemical bond on the golden film surface of carcinomebryonic antigen sensor
Specific immunity effect occurs for the carcinomebryonic antigen in fixed cancer embryo antibody and solution, and cancer embryo present in " crawl " solution resists
Original causes the mass loading on golden film surface to increase, and resonant frequency is reduced, therefore the frequency of surface acoustic wave reduces.Carcinomebryonic antigen passes
Input energy converter, output transducer and the Network Analyzer of sensor are all connected with and combined calculation machine carries out first frequency and the second frequency
Rate acquisition and analysis, the difference for finally obtaining second frequency and first frequency change with time figure, as shown in figure 11, it is known that, no
With (0g/ml, 0.5ng/ml, 1ng/ml, 5ng/ml, 10ng/ml) under carcinomebryonic antigen concentration, the difference of frequency changes with time,
When adding certain density carcinomebryonic antigen solution, with the passage in reaction time, the cancer embryo antibody in golden film is constantly adsorbed more
More carcinomebryonic antigens, second frequency become smaller, and the difference of frequency is negative value.It is infused when containing the carcinomebryonic antigen solution that concentration is 0.5ng/ml
When entering the detection zone of carcinomebryonic antigen sensor, biggish frequency shift (FS) occurs for the output signal of carcinomebryonic antigen sensor, and frequency becomes
Change amount (variable quantity i.e. between first frequency and second frequency) reaches 40KHz or so；It can learn that carcinomebryonic antigen sensor can
There is larger response to the carcinomebryonic antigen solution of low concentration (0.5ng/ml or less), and as carcinomebryonic antigen concentration increases, the second frequency
Rate and the difference of first frequency are also increasing.It is the difference and carcinomebryonic antigen concentration of first frequency and second frequency with reference to Figure 12, Figure 12
One specific embodiment linear relationship chart of constant logarithm；It obtains under different carcinomebryonic antigen concentration, first frequency and second frequency
Variable quantity is passed according to multiple frequency variations and known carcinomebryonic antigen concentration by the available carcinomebryonic antigen of linear analogue
Existing linear relationship between the frequency variation of sensor and the constant logarithm of carcinomebryonic antigen concentration.As shown in figure 12, the first frequency
There are a linear relationships for the logarithm of difference on the frequency and carcinomebryonic antigen concentration between rate and second frequency.It obtains difference on the frequency and cancer embryo is anti-
After the linear relationship of the constant logarithm of original content, when later use carcinomebryonic antigen sensor carries out carcinomebryonic antigen Concentration Testing, add
After adding on testing liquid to golden film, second frequency is obtained, the difference of second frequency Yu known first frequency is obtained, according to difference
The carcinomebryonic antigen concentration of testing liquid can be known with known linear relationship.
It is to be illustrated to preferable implementation of the invention, but the invention is not limited to the implementation above
Example, those skilled in the art can also make various equivalent variations on the premise of without prejudice to spirit of the invention or replace
It changes, these equivalent deformations or replacement are all included in the scope defined by the claims of the present application.
Claims (10)
1. a kind of carcinomebryonic antigen sensor, including substrate, which is characterized in that the carcinomebryonic antigen sensor further includes being arranged in institute
It states the piezoelectric material layer on substrate, the input energy converter that is arranged on the surface of the piezoelectric material layer and output transducer, set
The cancer embryo antibody setting the golden film between the input energy converter and the output transducer and being fixed in the golden film,
The input energy converter is used to for the rf signal of input being excited into surface acoustic wave, and the output transducer is used for will be received
Surface acoustic wave is converted to rf signal and is exported.
2. a kind of carcinomebryonic antigen sensor according to claim 1, which is characterized in that the input energy converter and/or defeated
Energy converter is interdigital transducer out.
3. a kind of carcinomebryonic antigen sensor according to claim 2, which is characterized in that the interdigital transducer has 20-
200 pairs of interdigital electrodes pair.
4. a kind of carcinomebryonic antigen sensor according to claim 2, which is characterized in that the interdigital width of the interdigital transducer
It spends range and finger spacing range is 1-50 microns.
5. a kind of carcinomebryonic antigen sensor according to any one of claims 1 to 4, which is characterized in that the substrate and/or
Piezoelectric material layer is piezoelectric monocrystal or piezoelectric membrane.
6. a kind of carcinomebryonic antigen sensor according to any one of claims 1 to 4, which is characterized in that the thickness of the golden film
Degree range is 100nm-200nm.
7. a kind of production method of carcinomebryonic antigen sensor, which is characterized in that be applied to as claimed in any one of claims 1 to 6
A kind of carcinomebryonic antigen sensor, comprising the following steps:
Production input energy converter and output transducer on piezoelectric material layer；
Make golden film on the piezoelectric material layer, the golden film setting the input energy converter and the output transducer it
Between；
Cancer embryo antibody is fixed in the golden film.
8. a kind of production method of carcinomebryonic antigen sensor according to claim 7, which is characterized in that utilize physical vapor
Deposition technique makes the golden film on the piezoelectric material layer.
9. a kind of production method of carcinomebryonic antigen sensor according to claim 7 or 8, which is characterized in that described in institute
Stating the method that cancer embryo antibody is fixed in golden film includes:
On the surface of the golden film, addition hydrosulphonyl silane alcoholic solution is reacted；
On the surface of the golden film, addition cysteamine ethanol solution is reacted；
On the surface of the golden film, addition cancer embryo antibody-solutions are reacted to fix cancer embryo antibody.
10. a kind of using a kind of carcinomebryonic antigen Concentration Testing side of carcinomebryonic antigen sensor as claimed in any one of claims 1 to 6
Method, which comprises the following steps:
The centre frequency of carcinomebryonic antigen sensor when being not added with testing liquid is set into first frequency；
Testing liquid is added in the golden film for being fixed with cancer embryo antibody to be reacted；
After fully reacting, the centre frequency for obtaining the carcinomebryonic antigen sensor is second frequency.
According to the line between the difference of the frequency of the first frequency and the second frequency and the constant logarithm of carcinomebryonic antigen concentration
Sexual intercourse obtains the carcinomebryonic antigen concentration of the testing liquid.
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