CN102520160A - Lamb wave immunosensor and manufacturing method thereof - Google Patents
Lamb wave immunosensor and manufacturing method thereof Download PDFInfo
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- CN102520160A CN102520160A CN2011103948549A CN201110394854A CN102520160A CN 102520160 A CN102520160 A CN 102520160A CN 2011103948549 A CN2011103948549 A CN 2011103948549A CN 201110394854 A CN201110394854 A CN 201110394854A CN 102520160 A CN102520160 A CN 102520160A
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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 gear shaping electrodes and a plurality of welding spot ports; a pair of electrode ports is arranged on each of the gear shaping 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 (MEMS) and biological immune sensing device crossing domain, specifically, relate to Lamb ripple and little magnetic bead fusion method mutually, 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 (MEMS) technology is one of current hi-tech development forward position; The high sensitivity and the immunoreactive specificity of sensor technology are merged; The signal that produces in the antigen-antibody reaction process, be transformed into electric signal through transducer, thereby antigen or antibody are carried out detection by quantitative.
Biology sensor converts the instrument that electric signal detects into to the biological substance sensitivity and with its concentration, is applied to fields such as clinical medicine food, industry and environment measuring gradually.Most important one type of immunosensor as biology sensor; Be to utilize the recognition function of antigen (antibody) antagonist (antigen) and the biology sensor that is developed into; Favored with its high degree of specificity, susceptibility and stability of identifying material, its appearance makes traditional immunoassay that very big variation take place.
Yet conventional immunity biosensor, for example surface plasma resonance immunosensor, quartz resonance type sensor are only realized single measurement, the molecular level fast of the array of very difficult realization sensor, high flux, 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.
The method for making of the Lamb wave device of the Lamb ripple immunosensor that another object of the present invention is to provide a kind of.
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 a magnet and a lower magnet on one, and said going up between magnet and the lower magnet is provided with a Lamb wave sensor, and said Lamb wave sensor top gummed has a pipeline gland; Said Lamb wave sensor comprises that one is provided with the silicon thin film structure of a plurality of sample cells; Said silicon thin film structure below is a conductive formation; Said conductive formation below is a piezoelectric material layer, is provided with by one deck IDT electrode layer on the said piezoelectric material layer, and said IDT electrode layer comprises some gear shaping electrodes and some solder joint ports; Be respectively arranged with the pair of electrodes port on described some gear shaping electrodes, cultivate underlined antibody, immune little magnetic bead and capture antibody in the said sample cell; Set a sample cell passage on the pipe surface that said pipeline gland and said Lamb wave sensor are fitted, the two ends of said sample cell passage offer a through hole respectively.
A kind of method for making of Lamb wave device, it may further comprise the 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 less than 3 μ m;
Step 2) get rid of photoresist: silicon chip at first cleans with the mixed liquor of acetone and alcohol, and soaked 5 minutes with deionized water the back, and oven dry afterwards is then at the two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine that silicon chip is made public,, afterwards, put into developer solution and corrode, generate required figure at photoresist layer at the figure of silicon chip surface generation template;
The step 4) corrode silicon dioxide: put into hydrofluoric acid solution to silicon chip, the corrode silicon dioxide layer makes the photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: put into potassium hydroxide solution to silicon chip, anisotropic etch silicon film former obtains the silicon thin film structure;
Step 6) is removed photoetching: put into acetone to silicon chip, remove photoresist;
Step 7) is removed silicon dioxide: put into hydrofluoric acid solution to silicon chip, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium/molybdenum: through using sputter, sputtered titanium/molybdenum on the back side of silicon thin film structure generates one deck conductive formation;
Step 9) sputter aluminium nitride: reactive sputtering one deck aluminium nitride on layer 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 the IDT figure;
Step 13) corroding electrode layer:, form the IDT electrode through the corroding electrode layer;
Step 14) is removed photoresist: remove photoresist, clean and oven dry.
A kind of method that detects carcinomebryonic antigen, it may further comprise the steps:
Step 1) is mixed analysans with immune little magnetic bead, the sample cell that injects the Lamb wave device is hatched jointly, and immune little magnetic bead surfaces is coated with certain carcinomebryonic antigen antibody;
Step 2) opens magnetic field, Lamb ripple immunosensor below, make immune magnetic ball be adsorbed to Lamb wave device surface, and form " immune magnetic ball-analysans-antibody " ternary complex jointly with the capture antibody on Lamb wave device surface;
Step 3) is opened magnetic field, Lamb ripple immunosensor top, makes the immune little magnetic bead that does not form " immune magnetic ball-analysans-antibody " ternary complex be adsorbed to Lamb wave device top;
Step 4) adds cleansing solution from through hole, remove the little magnetic bead of unconjugated immunity, simultaneously through Lamb ripple immunosensor synchro measure carcinomebryonic antigen concentration;
Step 5) adds regenerated liquid flushing Lamb wave device surface; " immune magnetic ball-analysans-antibody " the ternary complex form of destruction; Open magnetic field, Lamb ripple immunosensor top; Make immune little magnetic bead break away 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 present invention propose the Lamb ripple immunosensor method for making based on MEMS, be easy to and integrated circuit combines, be easy to realize the array of device, realize multi-channel detection; Adopt silicon chip as substrate in the present invention; Carry out the MEMS technological operation; Can on a device, produce one dimension or dimension sensor; Connect or the mode of flip chip bonding through gold ball bonding, link to each other sensor with electronic control system, solved the problem that the traditional optical sensor can not array.
2, utilize among the present invention MEMS technology to make film, its thickness is selected arbitrarily can be from 0.1 micron to 100 microns, satisfies the thickness requirement of transducer sensitivity to film fully.
3, the present invention utilizes the Lamb ripple immunosensor that MEMS technology is made, and is more suitable for mass production; Because the method that the immunomagnetic beads that the present invention proposes and Lamb wave sensor merge mutually; The fast enriching of immunomagnetic beads and the sensitivity of Lamb wave height combine together; The plurality of advantages that integrates both; Not only reduced analysis time, improved sensitivity and measuring accuracy, also made the 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 concentrates the back detection for the sample that is lower than detectability through the immunomagnetic beads enrichment, thereby changes detection limit indirectly, improves the sensitivity that detects, and avoids omission.
6, the Lamb ripple immunosensor based on MEMS of the present invention merges the immunomagnetic beads analytical approach, can realize immunoreactive array, high flux, large sample, the quick purpose of measuring.Compare with traditional immune analysis detection technique; Like enzyme immunoassay; Have high sensitivity, high specific, simple to operate, analysis speed is fast, 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 only is the general introduction of technical scheme of the present invention, understands technological means of the present invention in order can more to know, and can implement according to the content of instructions, below with preferred embodiment of the present invention and conjunction with figs. specify as after.
Description of drawings
Accompanying drawing described herein is used to provide further understanding of the present invention, constitutes the application's a part, and illustrative examples of the present invention and explanation thereof are used to explain the present invention, do not constitute improper qualification 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 accompanying drawing and combine embodiment, specify the present invention.
Embodiment 1:
Referring to Fig. 1, shown in Figure 2, a kind of Lamb ripple immunosensor, it comprises a magnet 1 and a lower magnet 11 on one, and said going up between magnet 1 and the lower magnet 11 is provided with a Lamb wave sensor 12, and said Lamb wave sensor 12 tops gummed has a pipeline gland 2; Further, in conjunction with shown in Figure 3, said Lamb wave sensor 12 comprises that one is provided with the silicon thin film structure 7 of a plurality of sample cells 701; Said silicon thin film structure 7 belows are a conductive formation 8; Said conductive formation 8 belows are a piezoelectric material layer 9, are provided with on the said piezoelectric material layer 9 by one deck IDT electrode layer, further; In conjunction with shown in Figure 4; Said IDT electrode layer comprises some gear shaping electrodes 10 and some solder joint port ones 3, is respectively arranged with pair of electrodes port one 4 on described some gear shaping electrodes 10, cultivates underlined antibody 3, immune little magnetic bead 4 and capture antibody 6 in the said sample cell 701; Further, in conjunction with shown in Figure 5, set a sample cell passage 18 on the pipe surface 17 that said pipeline gland 2 and said Lamb wave sensor 12 are fitted, the two ends of said sample cell passage 18 offer a through hole 19 respectively.
Preferably, the thickness at the film place of said silicon thin film structure 7 is between the 0.1 μ m-100 μ m.
Preferably, connect in succession through gold ball bonding between said electrode ports 14 and the solder joint port one 3.
Preferably, the height of the said piezoelectric material layer 9 of said gear shaping electrode 10 protrusions is 20nm-200nm.
Preferably, the distance between the surface of said sample cell passage 18 and the pipe surface 17 is 1 μ m-10 μ m.
Preferably, the bore dia of said through hole 19 is 0.2 μ m-10 μ m.
Embodiment 2:
Referring to shown in Figure 1, a kind of method for making of Lamb wave device, it may further comprise the 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 less than 3 μ m;
Step 2) get rid of photoresist: silicon chip at first cleans with the mixed liquor of acetone and alcohol, and soaked 5 minutes with deionized water the back, and oven dry afterwards is then at the two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine that silicon chip is made public,, afterwards, put into developer solution and corrode, generate required figure at photoresist layer at the figure of silicon chip surface generation template;
The step 4) corrode silicon dioxide: put into hydrofluoric acid solution to silicon chip, the corrode silicon dioxide layer makes the photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: put into potassium hydroxide solution to silicon chip, anisotropic etch silicon film former obtains silicon thin film structure 7;
Step 6) is removed photoetching: put into acetone to silicon chip, remove photoresist;
Step 7) is removed silicon dioxide: put into hydrofluoric acid solution to silicon chip, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium/molybdenum: through using sputter, sputtered titanium/molybdenum on the back side of silicon thin film structure 7 generates one deck conductive formation 8;
Step 9) sputter aluminium nitride: reactive sputtering one deck aluminium nitride on layer 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 the IDT figure;
Step 13) corroding electrode layer:, form the IDT electrode through the corroding electrode layer;
Step 14) is removed photoresist: remove photoresist, clean and oven dry.
Further, before step 9, also comprise following treatment step: be used for the circuit junction in for the conductive formation 8 that makes device and come out, use an other silicon chip to be placed on the said silicon thin film structure 7 and be used for the holding circuit junction.
Embodiment 3:
Referring to shown in Figure 1, a kind of method that detects carcinomebryonic antigen, it may further comprise the steps:
Step 1) is mixed analysans 5 with immune little magnetic bead 4, the sample cell 701 that injects Lamb wave device 12 is hatched jointly, and immune little magnetic bead 4 pan coatings have certain carcinomebryonic antigen antibody;
Step 2) opens magnetic field, Lamb ripple immunosensor below, make immune magnetic ball be adsorbed to Lamb wave device 12 surfaces, and form " immune magnetic ball-analysans-antibody " ternary complex jointly with the capture antibody 6 on Lamb wave device 12 surfaces;
Step 3) is opened magnetic field, Lamb ripple immunosensor top, makes the immune little magnetic bead 4 that does not form " immune magnetic ball-analysans-antibody " ternary complex be adsorbed to Lamb wave device 12 tops;
Step 4) adds cleansing solution from through hole 19, remove the little magnetic bead 4 of unconjugated immunity, simultaneously through Lamb ripple immunosensor synchro measure carcinomebryonic antigen concentration;
Step 5) adds regenerated liquid flushing Lamb wave device 12 surfaces; " immune magnetic ball-analysans-antibody " the ternary complex form of destruction; Open magnetic field, Lamb ripple immunosensor top; Make immune little magnetic bead 4 break away from Lamb wave devices 12 surfaces, and be washed and remove, reach the regeneration of Lamb ripple immunosensor.
The above is merely the preferred embodiments of the present invention, is not limited to the present invention, and for a person skilled in the art, the present invention can have various changes and variation.All within spirit of the present invention and principle, any modification of being done, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. Lamb ripple immunosensor; It is characterized in that: comprise a magnet on one (1) and a lower magnet (11); Said going up between magnet (1) and the lower magnet (11) is provided with a Lamb wave sensor (12), and said Lamb wave sensor (12) top gummed has a pipeline gland (2); Said Lamb wave sensor (12) comprises that one is provided with the silicon thin film structure (7) of a plurality of sample cells (701); Said silicon thin film structure (7) below is a conductive formation (8); Said conductive formation (8) below is a piezoelectric material layer (9); Said piezoelectric material layer (9) is gone up and is provided with by one deck IDT electrode layer; Said IDT electrode layer comprises some gear shaping electrodes (10) and some solder joint ports (13), is respectively arranged with pair of electrodes port (14) on described some gear shaping electrodes (10), cultivates underlined antibody (3), immune little magnetic bead (4) and capture antibody (6) in the said sample cell (701); Set a sample cell passage (18) on the pipe surface (17) that said pipeline gland (2) and said Lamb wave sensor (12) are fitted, the two ends of said sample cell passage (18) offer a through hole (19) respectively.
2. Lamb ripple immunosensor according to claim 1 is characterized in that: the thickness at the film place of said silicon thin film structure (7) is between the 0.1 μ m-100 μ m.
3. Lamb ripple immunosensor according to claim 1 is characterized in that: connect in succession through gold ball bonding between said electrode ports (14) and the solder joint port (13).
4. Lamb ripple immunosensor according to claim 1 is characterized in that: the height that said gear shaping electrode (10) protrudes said 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 said sample cell passage (18) and the 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 said 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, may further comprise the 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 less than 3 μ m;
Step 2) get rid of photoresist: silicon chip at first cleans with the mixed liquor of acetone and alcohol, and soaked 5 minutes with deionized water the back, and oven dry afterwards is then at the two-sided cementing agent and the photoresist of getting rid of of silicon chip;
Step 3) photoetching and development: utilize litho machine that silicon chip is made public,, afterwards, put into developer solution and corrode, generate required figure at photoresist layer at the figure of silicon chip surface generation template;
The step 4) corrode silicon dioxide: put into hydrofluoric acid solution to silicon chip, the corrode silicon dioxide layer makes the photoresist layer surfacial pattern transfer to silicon dioxide layer;
Step 5) corrosion silicon: put into potassium hydroxide solution to silicon chip, anisotropic etch silicon film former obtains silicon thin film structure (7);
Step 6) is removed photoetching: put into acetone to silicon chip, remove photoresist;
Step 7) is removed silicon dioxide: put into hydrofluoric acid solution to silicon chip, remove all silicon dioxide layers;
Step 8) splash-proofing sputtering metal titanium/molybdenum: through using sputter, sputtered titanium/molybdenum on the back side of silicon thin film structure (7) generates one deck conductive formation (8);
Step 9) sputter aluminium nitride: go up reactive sputtering one deck aluminium nitride in layer conductive formation (8), generate one deck piezoelectric material layer (9);
Step 10) sputtering electrode layer: go up sputtered aluminum, chromium or gold at 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 the IDT figure;
Step 13) corroding electrode layer:, form the IDT electrode through the corroding electrode layer;
Step 14) is removed photoresist: remove photoresist, clean and oven dry.
8. the method for Lamb wave device according to claim 7; It is characterized in that; Before step 9, also comprise following treatment step: be used for the circuit junction in for the conductive formation (8) that makes device and come out, use an other silicon chip to be placed on the said silicon thin film structure (7) and be used for the holding circuit junction.
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Cited By (4)
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| CN104406617A (en) * | 2014-12-10 | 2015-03-11 | 中国科学院苏州生物医学工程技术研究所 | Detachable sonic sensor signal test device |
| CN105181665A (en) * | 2015-09-18 | 2015-12-23 | 中国科学院苏州生物医学工程技术研究所 | Molecular dynamics test platform based on acousto-optical combination |
| CN107727845A (en) * | 2017-09-26 | 2018-02-23 | 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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| 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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