CN102103112B - Light addressing molecular imprinting array sensor for distinguishing residual pesticides - Google Patents
Light addressing molecular imprinting array sensor for distinguishing residual pesticides Download PDFInfo
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- CN102103112B CN102103112B CN 200910242771 CN200910242771A CN102103112B CN 102103112 B CN102103112 B CN 102103112B CN 200910242771 CN200910242771 CN 200910242771 CN 200910242771 A CN200910242771 A CN 200910242771A CN 102103112 B CN102103112 B CN 102103112B
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Abstract
The invention discloses a light addressing molecular imprinting array sensor for distinguishing residual pesticides, which is composed of a light addressing molecular imprinting array chip and a measuring cell of a light-addressable potentiometric sensor, wherein, the array chip based on a micro-electromechanical system (MEMS) technology is a silicon substrate microstructural array; a sol-gel molecular imprinting method is adopted on the surface of an array sensitive area of the chip to modify tetraethoxysilane, thus molecular imprinting sensitive membranes of various pesticides with the residues, such as organophosphorus, carbamic acid ester and the like can be distinguished; the measuring cell is made of organic glass, and comprises an upper cover, a measuring cell base and a seal rubber ring; the array chip can be arranged inside the measuring cell base interchangeably as a working electrode; an infrared light-emitting diode (LED) array is directly embedded outside the base by sealgum, and corresponds to the array chip; the inner side part corresponding to the working electrode array chip, of the upper cover is provided with a copper-based electroplate platinum (Pt) film as a counter electrode and an inlet and outlet pipeline; and the ring-shaped upper cover and the periphery of the measuring cell base are provided with screw threads, and the ring-shaped upper cover and the measuring cell base are sealed into the measuring cell through the seal rubber ring.
Description
Technical field
The invention belongs to the electrochemical sensor technical field that molecular imprinted membrane and light addressing detect, specifically relating to a kind of light addressing molecular engram sensor array of specific recognition remains of pesticide.
Background technology
The light addressing molecular engram sensor array of the specific recognition remains of pesticide that the present invention proposes is to adopt light addressing detection technique.This smooth addressing detection technique is based on Light Addressable Potentiometric Sensor (Light Addressable Potentiometric Sensor, LAPS), and this sensor is to be invented at late nineteen eighties by a molecular device company of California, USA.The advantage such as LAPS has strong, the high sensitivity of applied range, light addressing capability, higher stability, required sample is few, measurement range is wide, detection time is short becomes the nova of biosensor technique.The basic structure of LAPS as shown in Figure 1.Be insulated layer between semiconductor and the electrolyte solution and separate formation EIS structure.Regulating bias voltage makes semiconductor be in different bias states.Light source (such as LED or laser) through intensity modulated shines from the back side to this EIS structure, produces electron hole pair in semiconductor.When the surface that semiconductor contacts with insulation course was in spent condition, these electron hole pairs may enter depletion layer by diffusion.Electric field in the depletion layer with the light induced electron hole to separately, thereby in the loop, produce photogenerated current.When semiconductor surface was in the charge accumulated state, the electric field of semiconductor surface was zero, and electron hole pair can not separate at this, so photogenerated current is zero substantially.The responsive principle of LAPS is based on field effect makes device responsive to interface potential change between insulation course and electrolyte solution, but, when signals collecting, LAPS adopts the modulated beam of light irradiation, device is modulated by photocurrent the response of this potential change, and adopted phase-locked detection technique that response signal is detected.The typical response curve of LAPS as shown in Figure 2.When the concentration of test solution changed, response curve can be offset to the left or to the right.Can detect the variation of solution concentration by detecting this side-play amount.Because the existence of sensitive membrane, sensitive membrane adsorption one deck ion forms film potential, thereby causes the voltage at insulator and semiconductor two ends to produce certain skew.Therefore photocurrent-bias voltage curve also produces corresponding skew.Treat that the concentration of measured ion is relevant in the side-play amount of curve and the solution.Therefore can detect the concentration for the treatment of measured ion by the side-play amount of measuring curve.Measure empirical curve that the buffer solution of different pH values obtains as shown in Figure 2.Existing many scholars study on this basis its function are developed into enzyme LAPS, immune LAPS and biological LAPS from the quick LAPS of single H+ ion.
Biology sensor is comprised of recognition component and signal converter, recognition component is fixed on the surface of converter by rights, when testing molecule when recognition component is combined, produce a physics or chemical signal, converter with this signal convert to one can be quantitative output signal, realize the real time measure to testing molecule by monitoring output signal.The recognition component of conventional biosensor is made of biomolecule, such as enzyme, antibody, microorganism, tissue even complete organ; Converter has microelectrode, field effect transistor, optical fiber, thermistor and piezoelectric crystal etc.The present invention combines the molecular engram recognition component with light addressing transducer, new method and the new unit of the bionical biology sensor of research high stable, the molecule of the agricultural chemicals such as the residual organophosphorus of trial specificity Multiparameter, carbamate.
The high sensitivity of biology sensor and specific advantage are subject to extensive concern, but, the recognition component that is comprised of biomolecule exists has relatively high expectations to environment for use, is difficult to the intrinsic defectives such as long preservation, much is difficult for the obstacle that overcomes so that biology sensor runs in actual applications; Simultaneously, biomolecule derives from biological living, and preparation and purifying are loaded down with trivial details, expensive.Biology sensor face low stable, expensive, can not in pH value higher or on the low side and pyrosol, use, be difficult to do not have the corresponding series of problems such as identification target molecule with micro-processing technology compatibility, some analyte, become a key factor that affects its development.Therefore, obtaining cheap, stable recognition component, is one of key of further developing of biology sensor.By research and the understanding to antigen-antibody, enzyme and substrate reactions principle, scientist has courageously proposed the imagination by the synthetic analog antibody of chemical reaction, has started a brand-new technology-molecular imprinting.Molecular imprinting refers to prepare the polymkeric substance that a certain specific target molecules is had specific selectivity, and namely the process of molecularly imprinted polymer often is depicted as the technology of making " the artificial lock " of identifying " molecule key " visually.The molecular engram process was comprised of three steps: the first step, and function monomer and template molecule are mixed with out covalent complex, or form the addition compound product of non-covalent combination; Second step carries out polymerization with above-mentioned monomer-template complex (addition product), is frozen in high molecular three networks; The 3rd step removed template molecule from polymkeric substance.The space that former cause template molecule occupies will form the cavity that can remember formwork structure, size and other physics, chemical characteristic, can effectively optionally remove the molecule in conjunction with template (or analog).Molecularly imprinted polymer (molecularly imprinted polymers, MIPs) is consisted of bionical biology sensor as the bio-sensing functional membrane.
Engram technology is one of 20th century biology field three great discoveries.Discovery and the pcr gene amplification technique of it and restriction endonuclease have brought revolutionary progress for human bioengineering.Southern at first came analyzing DNA with this method in 1975, was called Southern blot; Alwine was used for RNA research with this method in 1979, was called Northern blot; The same year, Towbin etc. expanded to it again protein analysis, was named as Western blot (WBT), was called again Immunoblot (IBT), i.e. Western blotting.Can copy in theory the MIPs of any material, nearly 300 kinds of the MIPs of the inorganic ions of making, nucleic acid, protein even cell, and the emphasis that will study the engram technology of polymerization science focuses on, and utilizes computer software that molecule is carried out three-dimensional design.At organic sphere, also begin MIPs is combined with large-sized analytic instrument as sensitive element, nucleic acid, protein even cell are detected, and obtain gratifying achievement.Little by little the MIPs technology is applied to sensor in recent years: Panasyuk etc. utilize the grafting polymerization technique to prepare the molecular engram capacitive transducer at polypropylene screen and hydrophobicity gold electrode surfaces, have good selectivity.The people such as Kriz are fixed on the morphine molecularly imprinted polymer on the platinum electrode, measure the morphine that discharges by current method, and its detectable concentration scope is 0.01-1mg/L, and detection limit reaches 0.1ng/mL.The Piletsky of Ukraine utilizes cholesterol as template molecule, and hexadecane mercaptan adopts cyclic voltammetry that polymkeric substance is carried out electropolymerization as function monomer, and its sensing range is 15-60 μ M.Mullett etc. combine molecular imprinting and have developed a kind of theophylline sensor with Applications of surface plasmon resonance, measure concentration range and can reach 0.4-6g/L.Have in the situation of template molecule existence, in collosol-gelatum system, can form the micropore that template molecule is had the specific recognition ability.Bibliographical information is arranged the employing template molecule induce inorganic trace SiO
2Film is studied it to the selective problems of dopamine in the formation of electrode surface.Domestic analytical chemistry field is also very active to the research of MIPs, in Peking University, the molecular recognition mechanism of research MIPs, design, synthesize, estimate the novel molecular engram polymkeric substance with specificity and compatibility by the modern instrumental analysis means such as ultraviolet, infrared, chromatogram, specific surface, nuclear-magnetism and computer aided animation, and the condition of molecularly imprinted polymer preparation is optimized.They have proposed the concept of molecular engram originality, are used for describing the ability that a compound can produce high selectivity and high-affinity molecularly imprinted polymer.Also enlarging by the coordination of indirect molecular engram method and metallic ion in addition can be by the molecular range of trace, solves some little molecules and contains the trace problem of intramolecular hydrogen bond compound.Simultaneously, they also are applied to molecularly imprinted polymer the simulation of some native enzyme, constantly explore molecularly imprinted polymer in the application of catalytic field.Chemical defence research institute the 4th research institute is with the template molecule of methylphosphonic acid p-nitrophenyl ester as molecularly imprinted polymer, N-phenyl-benzamide is as the function base, synthesize the novel molecular engram analog antibody enzyme MIP-3 with " nanochannel " by being embedded into ZnO nano material, studied this enzyme to carboxylic acid p-nitrophenyl ester catalyzing hydrolysis dynamics; Result of study shows that this analoglike abzyme not only has good structure selectivity, and the ability of catalysis carboxyester hydrolysis improves greatly, for the application of synthetic molecules trace analogue enztme and nano material provides new way.
The molecular engram concept has existed for many years, still, to its experimental technique and application thereof, is in recent years along with developing rapidly of hyundai electronics and biotechnology just had considerable progress, becomes the focus of research.The stability of MIPs is higher than the bio-molecules recognition component far away, can obtain than high selectivity and specific polymkeric substance, is the ideal material that solves the bio-sensitive film poor stability.Therefore, the research of molecularly imprinted polymer is subject to people's attention, and research is to its optimal design, and exploitation MIP designs universal program.The at present research of MIPs biology sensor is except lacking MIP design universal program, also has MIP to lower, non-specific also higher, the more effective process for fixation etc. of target analytes compatibility, becoming the Main Bottleneck problem that MIPs is applied to biology sensor research, is scientist's problem anxious to be resolved of molecular chemistry and material science.And how research combines biosensor technique with intrinsic high stability advantage and the achievement in research of MIPs material, overcomes the difficult problem that MIPs exists at present.The combination of recognition component and converter mainly contains dual mode, and the one, make first polymer beads, be fixed to again on the converter, directly prepare blotting membrane in transducer face exactly in addition.The latter is the developing direction of molecular engram biomimetic sensor.The converter of molecular engram biomimetic sensor adopts the more of optical sensor or quartz crystal microbalance (QCM), adopts electrochemical sensor as the report of converter, and is gradually more and more in recent years.Also occur successively based on the molecular engram biomimetic sensor of potentiometric detection, also had some bibliographical informations at the TiO2 so-gel film of ISFET deposition shuttle based compound trace, study the problem of its selective response.But about light addressing molecular engram biomimetic sensor, there is not yet report.
Organophosphorus and carbamate are use maximum pesticide, germifuge, herbicide are arranged, and in vegetables, fruit, the grain, are often polluted, to the human and serious threat of other biological formation.The molecule that detects the agricultural chemicals such as residual organophosphorus, carbamate in food and the environment is pressing issues very.The standard detecting method of country is gas chromatography.Also often have enzyme to suppress method,, detect by the activity decreased of remains of pesticide inhibitory enzyme as catalyzer with enzyme, catalytic capability descends, and the pH of detected solution or colour developing are changed, and indirectly reflects persticide residue.Main Research Characteristics is the different enzymes of research and development.Also have for specific pesticide molecule, research and develop specific immune antiboidy material.The common issue with of these methods remains biomolecule and derives from biological living, and preparation and purifying are loaded down with trivial details, expensive.Biology sensor face low stable, expensive, can not in pH value higher or on the low side and pyrosol, use, be difficult to be subjected to all remainss of pesticide inhibition shortage specificitys, some analyte not to have the corresponding series of problems such as identification target molecule with micro-processing technology compatibility, enzyme, become a key factor that affects its development.In addition, said method also has a difficult problem that is difficult to overcome to be the nonreversibility of recognition reaction.These sensors can only disposablely use, and make troubles for practical popularization.The SiO that adopts the molecular engram preparation that organophosphorus and carbamate are had specific recognition
2Gel mould can overcome the problems referred to above.The research of this respect still is in the exploratory stage, and different function monomers, catalyzer, plasticizer materials and colloidal sol and bunching condition have very large research space.
Summary of the invention
The object of the present invention is to provide a kind of light addressing molecular engram sensor array of specific recognition remains of pesticide, to improve the defective that exists in the background.
For achieving the above object, the light addressing molecular engram sensor array of identification remains of pesticide provided by the invention includes the measuring cell base assembly, is contained in light addressing molecular engram array chip and the cover assembly of measuring cell base inboard; Wherein:
The measuring cell base is the circle cup of being made by organic glass;
The infrared LED matrix light source is being inlayed by central authorities in circle cup bottom, and a space of placing light addressing molecular engram array chip is established in LED matrix light source top;
The position of the chip that the measuring cell base is placed is equipped with the oxygen-free copper electrode, and picks out lead-in wire in base back surface, as the contact electrode of light addressing surveying work electrode;
The rim of a cup of measuring cell base is provided with external thread;
Light addressing molecular engram array chip, take the twin polishing silicon chip as substrate, surface and the back side at substrate all adopt each diversity chemical corrosion method to process N corresponding up and down hole, and each hole on surface represents a sensitive membrane sedimentary province, and each hole at the back side represents a light source region of acceptance;
The center in hole is corresponding with the center of infrared LED matrix light source respectively up and down;
The surface coverage thickness in each hole, surface of light addressing molecular engram array chip is respectively silicon dioxide layer, silicon nitride film and the tantalum pentoxide film of 50～100nm, again to modify sensitive membrane as the interface, remaining surface is as the insulation isolated area, be coated with first the thick silicon dioxide layer of 1 μ m, cover again silicon dioxide layer, silicon nitride film and the tantalum pentoxide film that is respectively 50～100nm with the surperficial the same thickness in hole;
Light addressing molecular engram array chip is prepared with the gold layer and is the contact electrode of working electrode chip;
The signal benchmark of medium position of light addressing molecular engram array chip, all the other N-1 position is as the sensitizing range of identification N-1 kind pesticide molecule;
Cover assembly is the dome of being made by organic glass;
The installed inside of dome has pair electrode, is substrate to electrode by oxygen-free copper garden sheet, surface preparation platinum film, draw from the sheet back side, oxygen-free copper garden lead-in wire as the light addressing measure to contact conductor;
Parallel with light addressing molecular engram array chip and the center is corresponding to the position of electrode;
Dome is provided with the import and export of two specimen;
The groove of annular is carved with to place O-ring seal in the dome inboard;
The dome mouth is provided with internal thread, and mutually combining with the external thread of measuring cell base rim of a cup consists of light addressing molecular engram sensor array.
In the light addressing molecular engram sensor array of described identification remains of pesticide, the infrared LED matrix light source is being inlayed by central authorities bottom the circle cup of measuring cell base, to seal with the circle cup end around the side of this matrix light source with fluid sealant, but must guarantee that the front (luminous point part) of light source and the back side (the electrode part of light source) are exposed to the outside.
In the light addressing molecular engram sensor array of described identification remains of pesticide, the position of infrared LED matrix light source and the chip position of placement are realized autoregistration.
In the light addressing molecular engram sensor array of described identification remains of pesticide, be placed with O-ring seal between the external thread of measuring cell base rim of a cup and the internal thread of dome mouth.
In the light addressing molecular engram sensor array of described identification remains of pesticide, the substrate of light addressing molecular engram array chip is that thickness is the twin polishing silicon chip of 5～8 Ω/cm less than 250 μ m, N-type " 100 ", resistivity.
In the light addressing molecular engram sensor array of described identification remains of pesticide, the oxidated layer thickness that the substrate surface hole covers is 100nm, and the thickness of silicon nitride film and tantalum pentoxide film is respectively 100nm; The oxidated layer thickness that the substrate remaining surface covers is 1 μ m, and the thickness of silicon nitride film and tantalum pentoxide film is respectively 100nm.
In the light addressing molecular engram sensor array of described identification remains of pesticide, the preparation of sensitive membrane, to adopt the method for sol-gel molecular engram to prepare the sol solution of sensitive membrane, the sol solution of catalyzer and template molecule will be added, repeatedly splash into again-extract, sol solution is fixed on the centre position and N-1 position of chip; Specific as follows:
1) take ethyl orthosilicate as body material, ethanol is solvent, and constant temperature stirs, prepare silicon acid-sol solution;
2) for the Si-OH hydroxy combining that can hang with the silicon gel of ethyl orthosilicate, select N-1 kind pesticide molecule as the molecular engram template molecule, respectively with 10: 1 volumetric molar concentration, molecule is sneaked into silica sol solution, with the silica sol solution of not mixing template molecule, form N kind molecular engram sol solution.And adding respectively hydrochloric acid as catalyzer, glycerine is plastifier, makes pH 1.7～1.8;
3) inject syringe with trace, repeat respectively N kind molecular engram sol solution to be dripped centre position and N-1 the position that is fixed on chip; Wherein the signal benchmark of silica sol of not mixing template molecule is fixed in the centre position, and all the other N-1 inject respectively a position silica sol that contains above-mentioned N-1 kind template molecule;
4) in 40-50 ℃ of freeze-day with constant temperature, finish gel process; Immerse the aqueous solution of methyl alcohol and sodium bicarbonate, the ultrasonic template molecule of removing is finally finished the immobilization of sensitive membrane again.
Characteristic of the present invention is:
1) Light Addressable Potentiometric Sensor and molecular engram sensor are integrated in one, both advantages and range of application are better brought into play;
2) the infrared LED matrix light source is embedded on the base, structurally guarantee the LAPS integrated chip pack into measuring cell just with base assembly effectively coupling and location, make chip and the easily autoregistration of infrared LED matrix light source, be convenient to the replacing of chip;
3) volume is little, and is compatible with general light address sensor test macro, can form the light addressing molecular engram sensing system of portable remains of pesticide.
Description of drawings
Fig. 1 is known LAPS system structural framework synoptic diagram;
Fig. 2 is the typical response curve of LAPS system shown in Figure 1;
Fig. 3 a is the synoptic diagram of the light addressing molecular engram sensor array of specific recognition remains of pesticide of the present invention;
Fig. 3 b is the cover assembly synoptic diagram of Fig. 3 a sensor;
Fig. 3 c is the lower bottom base assembly synoptic diagram of Fig. 3 a sensor;
Fig. 4 a is that the lower bottom base assembly of having inlayed LED matrix light source (application examples is 5 * 7 LED) is looked up and looked synoptic diagram;
Fig. 4 b is the cross-sectional view of having assembled the lower bottom base assembly of array chip;
Fig. 4 c is the schematic top plan view of having assembled the lower bottom base assembly of array chip;
Fig. 5 a is the cross-sectional view of array chip;
Fig. 5 b is the plan structure synoptic diagram of array chip;
Fig. 5 c is the sensitizing range elementary microstructure diagrammatic cross-section corresponding with sensitive area of array chip;
Fig. 6 is the test macro synoptic diagram of the light addressing molecular engram sensor array of specific recognition remains of pesticide;
Fig. 7 is array chip process flow diagram of the present invention; Wherein:
Fig. 7 a carries out an oxidation to silicon chip, and the front is got rid of photoresist and exposed;
Fig. 7 b is the positive etching field oxidation of back-protective, and each diversity etching silicon wafer forms pit;
Fig. 7 c is the fine and close thermal oxide layer of two-sided growth MOS level, the Si of LPCVD (low-pressure chemical vapor deposition method preparation)
3N
4Layer, the Ta of magnetron sputtering preparation
2O
5The pH sensitive layer;
Fig. 7 d is front photoresist protection, back side photoetching, and each diversity etching silicon wafer forms pit;
Fig. 7 e is back spatter Cr-Au back side Cr-Au conductive electrode.
Main label is among the figure:
1 cover assembly;
2 lower bottom base assemblies;
3 array chips;
4 organic glass loam cakes;
5 pairs of electrodes;
The contact conductor of 6 pairs of electrodes;
7 import and export pipeline;
8 O-ring seals;
The screw thread of 9 loam cakes;
10 organic glass bases;
The 11LED matrix light source;
The contact of 12 working electrodes and lead-in wire;
The screw thread of 13 organic glass bases;
14 silicon chips;
15 silicon chip surfaces have N recessed four ribs holes (application examples is 5);
16 silicon chip back sides have N recessed four ribs holes (application examples is 5);
17 chip back Cr-Au conductive electrodes;
18 chip reference potential districts;
Four sensitizing ranges of 19-22;
23SiO
2Insulation course;
24Si
3N
4Insulation course;
25Ta
2O
5Insulation;
26 molecular imprinted membranes;
27 lock-in amplifiers;
28 potentiostats; 29 Single Chip Microcomputer (SCM) system and led controller;
30 personal computers;
31 thick oxide layers;
32 photoresists;
C22, C24, C43, C62, C64 are respectively five luminous points of LED matrix light source.
Embodiment:
The working electrode of Light Addressable Potentiometric Sensor of the present invention is silicon based array microstructure electrode, but the molecular imprinted membrane of the agricultural chemicals such as the residual organophosphorus of the various specific recognition of electrode face finish, carbamate.The measuring cell of Light Addressable Potentiometric Sensor is made by organic glass, comprises loam cake, measuring cell base and O-ring seal (seeing also Fig. 3 a, Fig. 3 b, Fig. 3 c).
Measuring cell base assembly 2 is organic glass bases 10, its be shaped as circle cup (Fig. 3 a), it is of a size of:
Circle cup profile Φ 50mm * 25mm, rim of a cup has the external thread 13 of the high 5mm of M42mm;
One piece of 5 * 7 infrared LED matrix light source 11 inlayed by central authorities in circle cup bottom, this infrared LED matrix light source 11 is commercial prod, its physical dimension is 14.70mm * 19.80mm, spot diameter is Φ 1.9mm, spot distance 2.54mm, with the side and circle cup end sealing of fluid sealant with matrix light source, but must guarantee that the front (luminous point part) of light source and the back side (the electrode part of light source) are exposed to the outside, and guarantee no leakage;
35 luminous points in 5 * 7 infrared LED matrix light sources 11 are defined as Cxy, should select C22, C24, C43, C62, five luminous points of C64 in the use-case;
Above LED matrix light source 11, the space of a rectangular parallelepiped 19.85mm * 14.75mm * 1mm is arranged, just in time place light addressing molecular engram array chip;
Inboard at the measuring cell base, around the LED matrix light source 11, armouring the gold-plated oxygen-free copper electrode retaining collar of a rectangle, its physical dimension is 19.8mm * 14.7mm, endoporus is 14.75mm * 19.85mm, guarantees that the surface of light source of LED matrix light source and electrode retaining collar are embedded on the same plane, in addition, pick out the back side that lead-in wire is linked base from electrode retaining collar, the contact electrode 12 of the working electrode of measuring as the light addressing.
Dome profile Φ 50mm * 10mm has the internal thread 9 of the high 5mm of M42mm in the rim of a cup;
The oxygen-free copper electrode of inlaying one piece of gold,platinized in the central authorities of dome inside is as to electrode 5, and its physical dimension is Φ 30mm * 2mm, and the back side is by conductive adhesive, extraction electrode lead-in wire dome outside, become light addressing measurement to contact conductor 6;
Dome is inboard, is with a ring groove outside to electrode 5, wherein inlays O-ring seal 8, and it is of a size of Φ 47mm * 3mm;
Between to electrode 5 peripheries and O-ring seal 8, be provided with symmetrically stainless steel liquid inlet/outlet pipe road 7, be of a size of Φ 2mm * 15mm, tube wall 0.2mm with fluid sealant sealing all around, guarantees no leakage;
The external thread 13 of the rim of a cup of requirement measuring cell base is complementary with the internal thread 9 of dome mouth.After placing O-ring seal 8, again both are tightened, just consist of light addressing measuring cell.
Technological process such as the accompanying drawing 7 of light addressing molecular engram array chip 2:
Light addressing molecular engram array chip 2 is selected thickness less than 250 μ m, N-type＜100 〉, resistivity is that the twin polishing silicon chip 14 of 5-8 Ω/cm is substrate;
Array chip layout size: rectangular dies 19.8mm * 14.7mm, take the lower left corner of chip as reference point (0,0), the center of accepting illumination window at silicon chip surface sensitizing range and the back side is consistent with the center of luminous point, wherein C22 is (3.87,3.59), C24 is (3.87,8.62), C43 is (8.95,6.13), and C62 is (14.03,3.59), and C64 is (14.03,8.67); The window size of front domain is 1020 μ m * 1020 μ m, and the window size of back side domain is 1300 μ m * 1300 μ m;
According to the array chip schematic diagram of fabrication technology of Fig. 7 a-7e, the technological process of carrying out is as follows:
A) silicon chip 14 carries out an oxidation, the thick oxide layer 31 that the two-sided generation thickness of silicon chip 14 is 1 μ m, and the front is got rid of photoresist 32 and is exposed that (Fig. 7 is a);
B) the positive etching field oxide 31 of back-protective, etching depth is 10-20 μ m, and each diversity etching silicon wafer 14 forms pit 15 (Fig. 7 b);
C) the fine and close thermal oxide layer 23 of positive growth MOS level, its thickness is 100nm, and on the hot dioxide layer 23 of this densification chemical vapor deposition (LPCVD) Si
3N
4Layer 24, and at Si
3N
4The positive sputter Ta of layer 24
2O
5PH sensitive layer 25,23,24 and 25 be 100nm; The Cr-Au conductive electrode 17 (Fig. 7 c) of sputter 1 μ m overleaf;
D) front photoresist protection, back side photoetching, and each diversity etching silicon wafer 14 form pit 16 (Fig. 7 d), finish the MEMS technique of array chip;
E) the sensitive membrane sedimentary province on the array chip surface of having finished MEMS technique prepares molecular imprinted membrane 27.
Adopt each diversity chemical corrosion method to process deserved 5 holes, the degree of depth of surface imperfection be 10-20 μ m as the sensitive membrane sedimentary province, the degree of depth in 5 holes, the back side be about 150 μ m as the light source region of acceptance, cheating up and down floorage is 1mm * 1mm.Their center is corresponding with the center of 5 luminous points of C22, the C24 of 5 * 7 infrared LED array light sources, C43, C62, C64 respectively.
The surface coverage in 5 holes of rectangular dies the oxide layer 23 of 100nm, adds the silicon nitride film 24 of 100nm, adds 100nm tantalum pentoxide film 25, as the interface of modifying sensitive thin film; The remaining surface of chip is covered with the oxide layer of 1 μ m, adds the oxide layer 23 of 100nm, adds the silicon nitride film 24 of 100nm, adds 100nm tantalum pentoxide film 25, as the insulation isolated area.
The back side of chip does not have insulation course, and sputter Cr-Au conductive electrode 17 overleaf is as the contact electrode of working electrode chip.
The sensitive membrane preparation of light addressing molecular engram array chip, the signal benchmark in position, sensitizing range that the luminous point C43 of rectangular dies is corresponding, position, sensitizing range corresponding to all the other four luminous point C22, C24, C62, C64 is as pesticide molecule sensitizing ranges such as four kinds of organophosphoruss of identification, carbamates.The method of employing sol-gel molecular engram prepares the sol solution of sensitive membrane, will add the sol solution of catalyzer and template molecule, injects syringe with trace again, repeatedly splashes into-extraction method, sol solution is fixed on above-mentioned five positions of chip.Concrete grammar is as follows:
Take ethyl orthosilicate (TEOS) as body material, ethanol is solvent, with certain volumetric molar concentration, and adds a small amount of water, and 80 ℃ of constant temperature fully stirred 2 hours, prepare silicon acid-sol solution.
Select four kinds of pesticide molecules as the molecular engram template molecule, for example following four kinds of pesticide molecules:
1) flolimat (formal name used at school O, O-dimethyl-S-(N-methylamino formoxyl) thiophosphate, molecular formula C
5H
12NO
4PS).
2) metrifonate (formal name used at school dimethyl-2,2,2-three chloro-1-hydroxyethyl phosphates, molecular formula C
4H
8Cl
3O
4P).
3) acephatemet (formal name used at school O, S-dimethyl disulfide substituted phosphate, molecular formula C
2H
8NO
2PS).
4) DDVP (2,2-dichloroethylene methyl phosphorodithioate, molecular formula C
4H
7C
12O
4P).
With 10: 1 volumetric molar concentration, above-mentioned four kinds of molecules are sneaked into silica sol solution respectively, with the silica sol solution of not mixing template molecule, form five kinds of molecular engram sol solutions.And splashing into respectively several 0.05mol/L hydrochloric acid as catalyzer, several glycerine are plastifier, make pH at 1.7-1.8.
Inject syringe with trace, repeat (splash into-static 3 the second-draw again) ten times method, respectively five kinds of molecular engram sol solutions are dripped sensitizing range corresponding to luminous point C43 that is fixed on chip, and five positions of all the other four luminous point C22, C24, sensitizing range that C62, C64 are corresponding.The fixing signal benchmark of silica sol of not mixing template molecule in the position, sensitizing range that wherein luminous point C43 is corresponding, the silica sol that contains above-mentioned four kinds of template molecules is injected respectively in all the other four positions.
Inject the chip of colloidal sol, put into 50 ℃ of constant temperature ovens dry 3 days, finished gel process.
Fix the chip of gel, immersed the aqueous solution of methyl alcohol and sodium bicarbonate after 24 hours, renewed the ultrasonic 1-2 of liquid minute, removed template molecule.Finally finish the immobilization of sensitive membrane.
Claims (6)
1. light addressing molecular engram sensor array of identifying remains of pesticide includes the measuring cell base assembly, is contained in light addressing molecular engram array chip and the cover assembly as working electrode of measuring cell base inboard; Wherein:
The measuring cell base is the circle cup of being made by organic glass;
The infrared LED matrix light source is being inlayed by central authorities in circle cup bottom, and a space of placing light addressing molecular engram array chip is established in LED matrix light source top;
The position of the chip that the measuring cell base is placed is equipped with the oxygen-free copper electrode, and picks out lead-in wire in base back surface, as the contact electrode of light addressing surveying work electrode;
The rim of a cup of measuring cell base is provided with external thread;
Light addressing molecular engram array chip, take the twin polishing silicon chip as substrate, surface and the back side at substrate all adopt each diversity chemical corrosion method to process N corresponding up and down hole, and each hole on surface represents a sensitive membrane sedimentary province, and each hole at the back side represents a light source region of acceptance;
The center in hole is corresponding with the center of infrared LED matrix light source respectively up and down;
The surface coverage thickness in each hole, surface of light addressing molecular engram array chip is respectively silicon dioxide layer, silicon nitride film and the tantalum pentoxide film of 50～100nm, again to modify sensitive membrane as the interface, remaining surface is as the insulation isolated area, be coated with first the thick silicon dioxide layer of 1 μ m, cover again silicon dioxide layer, silicon nitride film and the tantalum pentoxide film that is respectively 50～100nm with the surperficial the same thickness in hole;
Light addressing molecular engram array chip is prepared with the gold layer and is the contact electrode of working electrode chip;
The signal benchmark of medium position of light addressing molecular engram array chip, all the other N-1 position is as the sensitizing range of identification N-1 kind pesticide molecule;
Cover assembly is the dome of being made by organic glass;
The installed inside of dome has pair electrode, is substrate to electrode by the oxygen-free copper disk, surface preparation platinum film, draw from the oxygen-free copper disk back side lead-in wire as the light addressing measure to contact conductor;
Parallel with light addressing molecular engram array chip and the center is corresponding to the position of electrode;
Dome is provided with the import and export of two specimen;
The groove of annular is carved with to place O-ring seal in the dome inboard;
The dome mouth is provided with internal thread, and mutually combining with the external thread of measuring cell base rim of a cup consists of light addressing molecular engram sensor array;
Wherein, the preparation of sensitive membrane is to adopt the method for sol-gel molecular engram to prepare the sol solution of sensitive membrane, will add the sol solution of catalyzer and template molecule, repeatedly splash into again-extract, sol solution is fixed on the centre position and N-1 position of chip; Specific as follows:
1) take ethyl orthosilicate as body material, ethanol is solvent, and constant temperature stirs, prepare silicon acid-sol solution;
2) for the Si-OH hydroxy combining that can hang with the silicon gel of ethyl orthosilicate, select N-1 kind pesticide molecule as the molecular engram template molecule, respectively with 10: 1 volumetric molar concentration, molecule is sneaked into silica sol solution, with the silica sol solution of not mixing template molecule, form N kind molecular engram sol solution, and add respectively hydrochloric acid as catalyzer, glycerine is plastifier, makes pH 1.7～1.8;
3) inject syringe with trace, repeat respectively N kind molecular engram sol solution to be dripped centre position and N-1 the position that is fixed on chip; Wherein the signal benchmark of silica sol of not mixing template molecule is fixed in the centre position, and all the other N-1 inject respectively a position silica sol that contains above-mentioned N-1 kind template molecule;
4) in 40-50 ℃ of freeze-day with constant temperature, finish gel process; Immerse the aqueous solution of methyl alcohol and sodium bicarbonate, the ultrasonic template molecule of removing is finally finished the immobilization of sensitive membrane again.
2. identify as claimed in claim 1 the light addressing molecular engram sensor array of remains of pesticide, wherein, the infrared LED matrix light source side that the central authorities of circle cup bottom inlay fluid sealant and the sealing of circle cup are outside the front and back of light source is exposed to.
3. the light addressing molecular engram sensor array of described identification remains of pesticide according to claim 1 and 2, wherein, the position of infrared LED matrix light source and the chip position of placement are realized autoregistration.
4. the light addressing molecular engram sensor array of described identification remains of pesticide according to claim 1 wherein, is placed with O-ring seal between the external thread of measuring cell base rim of a cup and the internal thread of dome mouth.
5. the light addressing molecular engram sensor array of described identification remains of pesticide according to claim 1, wherein, the substrate of light addressing molecular engram array chip is that thickness is less than 250 μ m, N-type＜100 〉, resistivity is the twin polishing silicon chip of 5～8 Ω/cm.
6. the light addressing molecular engram sensor array of described identification remains of pesticide according to claim 1, wherein, the oxidated layer thickness that the substrate surface hole covers is 100nm, the thickness of silicon nitride film and tantalum pentoxide film is respectively 100nm; The silicon dioxide layer thickness that the substrate remaining surface covers is 1 μ m, and the thickness of silicon dioxide layer, silicon nitride film and tantalum pentoxide film is respectively 100nm.
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| CN103675075A (en) * | 2013-11-27 | 2014-03-26 | 复旦大学 | Organophosphorus detection method based on microelectrode chip |
| JP6434744B2 (en) * | 2014-08-07 | 2018-12-05 | ローラス株式会社 | Semiconductor biosensor |
| CN104880453B (en) * | 2015-05-06 | 2017-11-03 | 华东理工大学 | The photoelectric synchronous method for sensing of solid nano passage based on dark-field imaging |
| CN115236064A (en) * | 2016-01-29 | 2022-10-25 | 梁庆耀 | Detection of organic chemicals |
| CN107478691A (en) * | 2017-08-08 | 2017-12-15 | 厦门英诺尔电子科技股份有限公司 | A kind of chemical sensor, preparation method and applications |
| CN109254046B (en) * | 2018-11-05 | 2021-01-12 | 济南大学 | Nitrofuran antibiotic sensor and preparation method thereof |
| CN110715964B (en) * | 2019-09-27 | 2022-08-19 | 西北工业大学 | Differential type optical addressing potential sensor |
| CN111103331B (en) * | 2019-12-27 | 2021-05-25 | 安徽芯淮电子有限公司 | Full-flexible heatable gas sensor and manufacturing method thereof |
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