CN102520160B - Lamb wave immunosensor and manufacturing method thereof - Google Patents
Lamb wave immunosensor and manufacturing method thereof Download PDFInfo
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- CN102520160B CN102520160B CN201110394854.9A CN201110394854A CN102520160B CN 102520160 B CN102520160 B CN 102520160B CN 201110394854 A CN201110394854 A CN 201110394854A CN 102520160 B CN102520160 B CN 102520160B
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Abstract
The invention discloses a Lamb wave immunosensor capable of performing array, high-flux, large-sample and quick measurement on immunoreaction. The Lamb wave immunosensor comprises an upper magnet and a lower magnet, wherein a Lamb wave sensor is arranged between the upper magnet and the lower magnet; a pipeline gland is glued above the Lamb wave sensor; the Lamb wave sensor comprises a silicon thin film structure with a plurality of sample cells; a conductive stratum is arranged below the silicon thin film structure; a piezoelectric material layer is arranged below the conductive stratum; an integrated device technology (IDT) electrode layer is arranged on the piezoelectric material layer, and comprises a plurality of interdigital electrodes and a plurality of welding spot ports; a pair of electrode ports is arranged on each of the interdigital electrodes; marking antibodies, immune micro magnetic balls and capturing antibodies are cultivated in the sample cells; a sample cell channel is formed on the pipeline surface of the pipeline gland, which is attached to the Lamb wave sensor; and a through hole is formed in each of two ends of the sample cell channel.
Description
Technical field
The invention belongs to MEMS (micro electro mechanical system) (MEMS) and biological immune sensing device crossing domain, specifically, relate to Lamb ripple and micro-magnetic bead phase fusion method, and the application of the method for making of Lamb wave device and this Lamb wave device.
Background technology
In recent years, array immunization sensor based on MEMS (micro electro mechanical system) (MEMS) technology is one of current hi-tech development forward position, the high sensitivity of sensor technology and immunoreactive specificity are merged, the signal producing in antigen-antibody reaction process, be transformed into electric signal through transducer, thereby antigen or antibody are quantitatively detected.
Biology sensor is responsive to biological substance and its concentration is converted to the instrument that electric signal detects, is applied to gradually the fields such as clinical medicine food, industry and environment measuring.Immunosensor is as the most important class of biology sensor, to utilize the recognition function of antigen (antibody) antagonist (antigen) and the biology sensor that is developed into, high degree of specificity, susceptibility and stability with its qualification material are favored, and its appearance has a very large change traditional immunoassay.
But conventional immunity biosensor, for example surface plasma resonance immunosensor, quartz resonance type sensor, only realize single measurement, is difficult to realize array, the high flux of sensor, the molecular level fast of large sample is measured.
Summary of the invention
For overcoming deficiency of the prior art, the object of the present invention is to provide a kind of Lamb ripple immunosensor that can realize immunoreactive array, high flux, large sample, Quick Measurement.
Another object of the present invention is the method for making of the Lamb wave device that a kind of Lamb ripple immunosensor is provided.
A further object of the present invention is to provide a kind of detection method of Lamb ripple immunosensor.
For realizing above-mentioned technical purpose, reach above-mentioned technique effect, the present invention has adopted following technical scheme:
A kind of Lamb ripple immunosensor, it comprises magnet and a lower magnet on one, is provided with a Lamb wave sensor between described upper magnet and lower magnet, described Lamb wave sensor top gummed has a pipeline gland; Described Lamb wave sensor comprises that one is provided with the silicon thin film structure of multiple sample cells, described silicon thin film structure below is a conductive formation, described conductive formation below is a piezoelectric material layer, on described piezoelectric material layer, arrange by one deck IDT electrode layer, described IDT electrode layer comprises some interdigital electrodes and some solder joint ports, in described some interdigital electrodes, be respectively arranged with pair of electrodes port, in described sample cell, cultivate and have labelled antibody, immune micro-magnetic bead and capture antibody; On the pipe surface of described pipeline gland and the laminating of described Lamb wave sensor, set a sample cell passage, the two ends of described sample cell passage offer respectively a through hole.
A method for making for Lamb wave device, it comprises the following steps:
Step 1) is prepared silicon chip: get one 3 inches, thick, the two-sided oxidation of 380 μ m, surface thickness and change P type (100) silicon chip that is less than 3 μ m;
Step 2) get rid of photoresist: first silicon chip cleans with the mixed liquor of acetone and alcohol, soaks 5 minutes with deionized water afterwards, dries afterwards, then at two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine to expose to silicon chip, generate the figure of template at silicon chip surface, afterwards, put into developer solution and corrode, generate required figure at photoresist layer;
Step 4) corrode silicon dioxide: silicon chip is put into hydrofluoric acid solution, and corrode silicon dioxide layer, makes photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: silicon chip is put into potassium hydroxide solution, and anisotropic etch silicon film former, obtains silicon thin film structure;
Step 6) is removed photoetching: silicon chip is put into acetone, photoresist is removed;
Step 7) is removed silicon dioxide: silicon chip is put into hydrofluoric acid solution, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium and molybdenum: by using sputter, sputtered titanium and molybdenum on the back side of silicon thin film structure, generate one deck conductive formation;
Step 9) sputter aluminium nitride: reactive sputtering one deck aluminium nitride in conductive formation, generates one deck piezoelectric material layer;
Step 10) sputtering electrode layer: sputtered aluminum, chromium or gold on piezoelectric material layer, generate electrode layer;
Step 11) is got rid of photoresist: on electrode, get rid of one deck photoresist;
Step 12) exposure and development: on photoresist layer, form IDT figure;
Step 13) corroding electrode layer: by corroding electrode layer, form IDT electrode;
Step 14) is removed photoresist: photoresist is removed, cleaned and dry.
A method that detects carcinomebryonic antigen, it comprises the following steps:
Step 1) is mixed analysans with immune micro-magnetic bead, the sample cell that injects Lamb wave device is hatched jointly, and immune micro-magnetic bead surfaces is coated with certain carcinomebryonic antigen antibody;
Step 2) open magnetic field, Lamb ripple immunosensor below, make biomolecular be adsorbed to Lamb wave device surface, and jointly form " biomolecular-analysans-antibody " ternary complex with the capture antibody on Lamb wave device surface;
Step 3) is opened magnetic field, Lamb ripple immunosensor top, makes the micro-magnetic bead of immunity that does not form " biomolecular-analysans-antibody " ternary complex be adsorbed to Lamb wave device top;
Step 4) adds cleansing solution from through hole, removes the micro-magnetic bead of unconjugated immunity, simultaneously by Lamb ripple immunosensor synchro measure carcinomebryonic antigen concentration;
Step 5) adds regenerated liquid to rinse Lamb wave device surface, destroy " biomolecular-analysans-antibody " ternary complex form, open magnetic field, Lamb ripple immunosensor top, make immune micro-magnetic bead depart from Lamb wave device surface, and be washed and remove, reach the regeneration of Lamb ripple immunosensor.
Compared with prior art, the present invention has following beneficial effect:
1, the Lamb ripple immunosensor method for making based on MEMS that the present invention proposes, is easy to and integrated circuit combination, is easy to realize the array of device, realizes multi-channel detection; Adopt in the present invention silicon chip as substrate, carry out MEMS technological operation, can on a device, produce one dimension or dimension sensor, connect or the mode of flip chip bonding by gold ball bonding, sensor is connected with electronic control system, has solved the problem that traditional optical sensor can not array.
2, utilize in the present invention MEMS technique to make film, its thickness is selected arbitrarily can be from 0.1 micron to 100 microns, meets the thickness requirement of transducer sensitivity to film completely.
3, the present invention utilizes the Lamb ripple immunosensor that MEMS technique is made, and is more suitable for mass production; The method that the immunomagnetic beads proposing due to the present invention merges mutually with Lamb wave sensor, the fast enriching of immunomagnetic beads and the sensitivity of Lamb wave height combine together, integrate both plurality of advantages, not only reduce analysis time, improved sensitivity and measuring accuracy, also make mensuration process become simply, easily be automated.
4, the immunomagnetic beads carrier specificity of the Lamb ripple immunosensor based on MEMS of the present invention is good, and solid-liquid can be easier to separate through magneticaction, and elution requirement is more environmentally friendly.
5, the Lamb ripple immunosensor based on MEMS of the present invention detects after immunomagnetic beads enrichment is concentrated for the sample lower than detectability, thereby indirectly changes detection limit, improves the sensitivity detecting, and avoids undetected.
6, the Lamb ripple immunosensor based on MEMS of the present invention, merges immunomagnetic beads analytical approach, can realize the object of immunoreactive array, high flux, large sample, Quick Measurement.Compare with traditional immune analysis detection technique, as enzyme immunoassay, there is high sensitivity, high specific, simple to operate, analysis speed is fast, the advantage such as cheap, and easily be automated operation, will be used widely in fields such as clinical diagnosis, health care, environmental monitoring, food securities.
Above-mentioned explanation is only the general introduction of technical solution of the present invention, in order to better understand technological means of the present invention, and can be implemented according to the content of instructions, below with preferred embodiment of the present invention and coordinate accompanying drawing to be described in detail as follows.
Brief description of the drawings
Accompanying drawing described herein is used to provide a further understanding of the present invention, forms the application's a part, and schematic description and description of the present invention is used for explaining the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings:
Fig. 1 is the schematic diagram of Lamb ripple immunosensor of the present invention.
Fig. 2 is the structural representation of Lamb ripple immunosensor of the present invention.
Fig. 3 is the vertical view of Lamb wave device of the present invention.
Fig. 4 is the upward view of Lamb wave device of the present invention.
Fig. 5 is the stereographic map of pipeline gland of the present invention.
Embodiment
Below with reference to the accompanying drawings and in conjunction with the embodiments, describe the present invention in detail.
embodiment 1:
Shown in Fig. 1, Fig. 2, a kind of Lamb ripple immunosensor, it comprises magnet 1 and a lower magnet 11 on one, between described upper magnet 1 and lower magnet 11, is provided with a Lamb wave sensor 12, described Lamb wave sensor 12 top gummeds have a pipeline gland 2, further, shown in Fig. 3, described Lamb wave sensor 12 comprises that one is provided with the silicon thin film structure 7 of multiple sample cells 701, described silicon thin film structure 7 belows are a conductive formation 8, described conductive formation 8 belows are a piezoelectric material layer 9, on described piezoelectric material layer 9, arrange by one deck IDT electrode layer, further, shown in Fig. 4, described IDT electrode layer comprises some interdigital electrodes 10 and some solder joint port ones 3, in described some interdigital electrodes 10, be respectively arranged with pair of electrodes port one 4, the interior cultivation of described sample cell 701 has labelled antibody 3, the micro-magnetic bead 4 of immunity and capture antibody 6, further, shown in Fig. 5, on the pipe surface 17 that described pipeline gland 2 and described Lamb wave sensor 12 are fitted, set a sample cell passage 18, the two ends of described sample cell passage 18 offer respectively a through hole 19.
Preferably, the thickness at the film place of described silicon thin film structure 7 is between 0.1 μ m-100 μ m.
Preferably, between described electrode ports 14 and solder joint port one 3, connect in succession by gold ball bonding.
Preferably, to protrude the height of described piezoelectric material layer 9 be 20nm-200nm to described interdigital electrode 10.
Preferably, the distance between the surface of described sample cell passage 18 and pipe surface 17 is 1 μ m-10 μ m.
Preferably, the bore dia of described through hole 19 is 0.2 μ m-10 μ m.
embodiment 2:
Shown in Figure 1, a kind of method for making of Lamb wave device, it comprises the following steps:
Step 1) is prepared silicon chip: get one 3 inches, thick, the two-sided oxidation of 380 μ m, surface thickness and change P type (100) silicon chip that is less than 3 μ m;
Step 2) get rid of photoresist: first silicon chip cleans with the mixed liquor of acetone and alcohol, soaks 5 minutes with deionized water afterwards, dries afterwards, then at two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine to expose to silicon chip, generate the figure of template at silicon chip surface, afterwards, put into developer solution and corrode, generate required figure at photoresist layer;
Step 4) corrode silicon dioxide: silicon chip is put into hydrofluoric acid solution, and corrode silicon dioxide layer, makes photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: silicon chip is put into potassium hydroxide solution, and anisotropic etch silicon film former, obtains silicon thin film structure 7;
Step 6) is removed photoetching: silicon chip is put into acetone, photoresist is removed;
Step 7) is removed silicon dioxide: silicon chip is put into hydrofluoric acid solution, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium and molybdenum: by using sputter, sputtered titanium and molybdenum on the back side of silicon thin film structure 7, generate one deck conductive formation 8;
Step 9) sputter aluminium nitride: reactive sputtering one deck aluminium nitride in conductive formation 8, generates one deck piezoelectric material layer 9;
Step 10) sputtering electrode layer: sputtered aluminum, chromium or gold on piezoelectric material layer 9, generate electrode layer;
Step 11) is got rid of photoresist: on electrode layer, get rid of one deck photoresist;
Step 12) exposure and development: on photoresist layer, form IDT figure;
Step 13) corroding electrode layer: by corroding electrode layer, form IDT electrode;
Step 14) is removed photoresist: photoresist is removed, cleaned and dry.
Further, before step 9, also comprise following treatment step: in order to make to come out for circuit junction in the conductive formation 8 of device, use an other silicon chip to be placed in described silicon thin film structure 7 for the protection of circuit junction.
embodiment 3:
Shown in Figure 1, a kind of method that detects carcinomebryonic antigen, it comprises the following steps:
Step 1) is mixed analysans 5 with immune micro-magnetic bead 4, the sample cell 701 that injects Lamb wave device 12 is hatched jointly, and immune micro-magnetic bead 4 pan coatings have certain carcinomebryonic antigen antibody;
Step 2) open magnetic field, Lamb ripple immunosensor below, make biomolecular be adsorbed to Lamb wave device 12 surfaces, and jointly form " biomolecular-analysans-antibody " ternary complex with the capture antibody 6 on Lamb wave device 12 surfaces;
Step 3) is opened magnetic field, Lamb ripple immunosensor top, makes the micro-magnetic bead 4 of immunity that does not form " biomolecular-analysans-antibody " ternary complex be adsorbed to Lamb wave device 12 tops;
Step 4) adds cleansing solution from through hole 19, removes the micro-magnetic bead 4 of unconjugated immunity, simultaneously by Lamb ripple immunosensor synchro measure carcinomebryonic antigen concentration;
Step 5) adds regenerated liquid to rinse Lamb wave device 12 surfaces, destroy " biomolecular-analysans-antibody " ternary complex form, open magnetic field, Lamb ripple immunosensor top, make immune micro-magnetic bead 4 depart from Lamb wave device 12 surfaces, and be washed and remove, reach the regeneration of Lamb ripple immunosensor.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (8)
1. a Lamb ripple immunosensor, it is characterized in that: comprise magnet on one (1) and a lower magnet (11), between described upper magnet (1) and lower magnet (11), be provided with a Lamb wave sensor (12), described Lamb wave sensor (12) top gummed has a pipeline gland (2), described Lamb wave sensor (12) comprises that one is provided with the silicon thin film structure (7) of multiple sample cells (701), described silicon thin film structure (7) below is a conductive formation (8), described conductive formation (8) below is a piezoelectric material layer (9), on described piezoelectric material layer (9), be provided with one deck IDT electrode layer, described IDT electrode layer comprises some interdigital electrodes (10) and some solder joint ports (13), in described some interdigital electrodes (10), be respectively arranged with pair of electrodes port (14), in described sample cell (701), cultivate and have labelled antibody (3), the micro-magnetic bead of immunity (4) and capture antibody (6), on the pipe surface (17) of described pipeline gland (2) and described Lamb wave sensor (12) laminating, set a sample cell passage (18), the two ends of described sample cell passage (18) offer respectively a through hole (19).
2. Lamb ripple immunosensor according to claim 1, is characterized in that: the thickness at the film place of described silicon thin film structure (7) is between 0.1 μ m-100 μ m.
3. Lamb ripple immunosensor according to claim 1, is characterized in that: between described electrode ports (14) and solder joint port (13), connect in succession by gold ball bonding.
4. Lamb ripple immunosensor according to claim 1, is characterized in that: the height that described interdigital electrode (10) is protruded described piezoelectric material layer (9) is 20nm-200nm.
5. Lamb ripple immunosensor according to claim 1, is characterized in that: the distance between the surface of described sample cell passage (18) and pipe surface (17) is 1 μ m-10 μ m.
6. Lamb ripple immunosensor according to claim 1, is characterized in that: the bore dia of described through hole (19) is 0.2 μ m-10 μ m.
7. a method of making the Lamb wave device of Lamb ripple immunosensor as claimed in claim 1, is characterized in that, comprises the following steps:
Step 1) is prepared silicon chip: get one 3 inches, thick, the two-sided oxidation of 380 μ m, surface thickness and change P type (100) silicon chip that is less than 3 μ m;
Step 2) get rid of photoresist: first silicon chip cleans with the mixed liquor of acetone and alcohol, soaks 5 minutes with deionized water afterwards, dries afterwards, then at two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine to expose to silicon chip, generate the figure of template at silicon chip surface, afterwards, put into developer solution and corrode, generate required figure at photoresist layer;
Step 4) corrode silicon dioxide: silicon chip is put into hydrofluoric acid solution, and corrode silicon dioxide layer, makes photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: silicon chip is put into potassium hydroxide solution, and anisotropic etch silicon film former, obtains silicon thin film structure (7);
Step 6) is removed photoetching: silicon chip is put into acetone, photoresist is removed;
Step 7) is removed silicon dioxide: silicon chip is put into hydrofluoric acid solution, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium and molybdenum: by using sputter, sputtered titanium and molybdenum on the back side of silicon thin film structure (7), generate one deck conductive formation (8);
Step 9) sputter aluminium nitride: at the upper reactive sputtering one deck aluminium nitride of conductive formation (8), generate one deck piezoelectric material layer (9);
Step 10) sputtering electrode layer: at piezoelectric material layer (9) upper sputtered aluminum, chromium or gold, generate electrode layer;
Step 11) is got rid of photoresist: on electrode layer, get rid of one deck photoresist;
Step 12) exposure and development: on photoresist layer, form IDT figure;
Step 13) corroding electrode layer: by corroding electrode layer, form IDT electrode;
Step 14) is removed photoresist: photoresist is removed, cleaned and dry.
8. the method for making Lamb wave device according to claim 7; it is characterized in that; before step 9, also comprise following treatment step: in order to make to come out for circuit junction in the conductive formation (8) of device, use an other silicon chip to be placed on described silicon thin film structure (7) upper for the protection of circuit junction.
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| CN104406617B (en) * | 2014-12-10 | 2017-02-22 | 中国科学院苏州生物医学工程技术研究所 | Detachable sonic sensor signal test device |
| CN105181665B (en) * | 2015-09-18 | 2018-09-14 | 中国科学院苏州生物医学工程技术研究所 | Based on molecular dynamics test platform associated with acousto-optic |
| CN107727845B (en) * | 2017-09-26 | 2019-09-10 | 中国科学院苏州生物医学工程技术研究所 | Lamb wave sensor, biological detection chip and fast screening system |
| CN111579594A (en) * | 2020-05-27 | 2020-08-25 | 上海交通大学 | Application of room temperature gas sensor in nerve agent detection |
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