CN104345037A - Optical detection sensor for microcystin - Google Patents
Optical detection sensor for microcystin Download PDFInfo
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- CN104345037A CN104345037A CN201410617885.XA CN201410617885A CN104345037A CN 104345037 A CN104345037 A CN 104345037A CN 201410617885 A CN201410617885 A CN 201410617885A CN 104345037 A CN104345037 A CN 104345037A
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- microcystin
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Abstract
The invention discloses an optical detection sensor for microcystin. The optical detection sensor comprises a linear polarization light source, a 45-degree/45-degree/90-degree glass prism, a sensing chip and a spectrum analyzer and is characterized in that a silicon circulation tank is arranged on the sensing chip; the sensing chip comprises a glass sheet, a gold layer coated on the surface of one side of the glass sheet as well as a microcystin molecularly imprinted membrane modified on the surface of the gold layer. The optical detection sensor disclosed by the invention has the beneficial effects that firstly, a molecular imprinting technique is adopted for forming a sensitive layer of the sensor, so that specific recognition and adsorption efficiency for the microcystin are enhanced, the specificity of detection means is improved, and the detection time is shortened; secondly, by the use of a surface plasma optical detection technology, the sensitivity is improved, and the performance of the sensor is improved; thirdly, the whole sensor is simple and quick to operate, is low in requirement on operators, and provides effective help for highly sensitive detection of the microcystin.
Description
Technical field
The present invention relates to a kind of Microcystin optical detecting sensor.
Background technology
In Middle And Lower Reaches of Changjiang River is one of area of China's freshwater lake integrated distribution, and in the past few decades, the lake ecosystem of mankind's activity on this area creates great impact, the eutrophication day by day of the most outstanding is suburb lakes.Receive the nutriments such as too much nitrogen and phosphorus in lake, reservoir and river, the ecologic structure of water body and function changed, cause algae particularly blue-green algae distorted proliferation growth and there is blue-green alga bloom phenomenon.The frequent generation of the harmful algal blooms caused along with the aggravation of body eutrophication has become the environmental problem of common concern.When blue-green alga bloom is serious, not only affects the sense organ of people, destroy aquatic ecosystem in a healthy and balanced way, and frustule break after the drinking water safety of various microcystins to humans and animals that discharge constitute serious threat.In the various Microcystins found, Microcystin is that known at present a kind of in blue-green alga bloom pollutes, the frequency of occurrences is the highest, generation is maximum and the most serious Microcystin that works the mischief, therefore very necessary to the detection of Microcystin.
At present, to detection method mainly chemical analysis and the immunological detection of Microcystin.Using more in chemical detection method is high performance liquid chromatography (HPLC) technology.HPLC technology generally adopts positive or reverse-phase chromatography contratoxin to be separated, then carry out ultraviolet (UV), fluorescence (FL) or chemiluminescence (CL) to detect, the separation of Microcystin, qualification can be widely used in and quantitatively detect.The hold-up time of tested toxin and normaltoxin is compared and can identify tested toxin, both peak areas is compared and can carry out accurate quantification to it.But HPLC method and technology content is high, expensive; The pure toxin of standard or the retention of certain toxin under same experimental conditions must be had in addition.Due to the widespread use of plastic additive in our times plastic products, gather some plastic additive in the plastic containers of water sample and HPLC can be disturbed the detection of Microcystin.Immunological detection is enzyme-linked immuno assay (ELISA) mainly, apply this type of analytical approach screening toxin advantage be highly sensitive, process gross sample time analysis speed fast, principle of work is simple.Under the prerequisite possessing microcystin monoclonal antibody, the pure toxin of standard and associated reagents, during the kit of especially commodity in use, ELISA method be one very easy, efficiently, method fast.Shortcoming can not differentiate toxin very well, there will be false positive reaction.
Chinese patent 201310712994.5 discloses a kind of quick-check sensor of Microcystin, use photochemical polymerization method or electrochemical polymerization to carry out molecular engram film synthesis, improve binding capacity, reduce time delay and strengthen specific adsorption ability; Meanwhile, use Lamb wave piezoelectric membrane as the effector of sensor, improve transducer sensitivity, increase stability and reduce costs.Thus enhancing sensor performance, achieve the trace detection to Microcystin, establish high sensitivity, fast, the Microcystin detection means of low cost, low operation requirements.
Chinese patent 201310713093.8 discloses a kind of Microcystin piezoelectric detection sensor based on molecular imprinting, use photochemical polymerization method or electrochemical polymerization to carry out molecular engram film synthesis, improve binding capacity, reduce time delay and strengthen specific adsorption ability; Meanwhile, use quartz crystal as the effector of sensor, improve transducer sensitivity, increase stability and reduce costs.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of Microcystin optical detecting sensor, and for the sensing chip of Microcystin optical detecting sensor.
The present invention is achieved through the following technical solutions:
A kind of Microcystin optical detecting sensor, comprise linear polarization light source, 45 °/45 °/90 ° glass prisms, sensing chip, spectroanalysis instruments, described sensing chip is provided with silica gel circulation groove, described sensing chip comprises glass sheet, is plated in the layer gold of glass sheet single side surface and is modified at the Microcystin molecular engram film on layer gold surface, Microcystin molecular engram film has to Microcystin specific recognition and equally distributed micropore.
As a kind of scheme, adopt rubbing method that Microcystin molecular engram film is modified at layer gold surface.Rubbing method is a kind of main classic method, first grade or nano level molecularly imprinted polymer particle is prepared, again polymer beads is embedded in gel or film, adopt rotary coating or the obtained identification layer of spraying coating, the problem that this method is brought be extend the sensor response time, non-specific adsorption is produced to target molecule and binding capacity low.
As a kind of preferred version, thermal polymerization Microcystin molecular engram film is adopted to be modified at layer gold surface.This method directly can obtain identification layer in position, avoids the drawback of classic method, and reproducible, environmental friendliness.
Further, described hot polymerization is combined into: gold-plated glass sheet is used oxygen gas plasma washer cleaning 2-4 minute, soak more than 12 hours in the ethanolic solution of 50-1000mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.1-1M and the N-hydroxy-succinamide of 0.01-0.1M 1 hour; After taking out directly in immersion 100-500mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 0.1-200umol Microcystin, 1-2000umol methacrylic acid and 2-2000umol trimethylol-propane trimethacrylate and add the mixing of 5-10mL dimethyl sulphoxide solution, nitrogen treatment more than 10 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 18 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 12 time more than, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
In the present invention, detect specificity for sensor, the needs of detection speed, the present invention proposes the sensitive layer using molecular imprinting as sensor, the principle of molecular imprinting is that template molecule and function monomer are in dispersion medium, rely on interaction force and space steric effect etc., form the compound of Reversible binding, add crosslinking chemical and initiating agent initiated polymerization formation porous functional material again, and template molecule is wrapped in wherein regularly, synthesize rear ad hoc approach template molecule is removed, thus acquisition can be complementary with template molecule, and there is the three-dimensional cavity of special identification function, thus fast, high specific in conjunction with template molecule.
In the present invention, for sensor to high sensitivity, easy needs, the present invention proposes to use surface plasma resonance sensor to be converter, this easy, highly sensitive optical sensor can variations in refractive index sensor surface being detected of rapid sensitive, and device is stablized, antijamming capability is strong.Caused the change of surface refractive index by the molecular engram film in effector finishing in conjunction with Microcystin, thus cause effector signal to change, utilize signal acquiring system acquisition and recording signal intensity, just can detect the Microcystin in sample.
Compared with prior art, the invention has the beneficial effects as follows:
1, have employed the sensitive layer of molecular imprinting as sensor, enhance the specific recognition to Microcystin, adsorption efficiency, improve the specificity of detection means and shorten detection time;
2, employ surface plasma optical detective technology, improve sensitivity, improve the performance of sensor;
3, whole sensor operations is simple, quick, requires low to operating personnel, for realizing providing strong help to the highly sensitive detection of Microcystin.
Accompanying drawing explanation
Fig. 1 is the structural representation of Microcystin optical detecting sensor.
Fig. 2 is the structural representation of sensing chip.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described:
As depicted in figs. 1 and 2, a kind of Microcystin optical detecting sensor, comprise linear polarization light source, 45 °/45 °/90 ° glass prisms 4, sensing chip 7, spectroanalysis instruments 6, sensing chip 7 is provided with silica gel circulation groove 5, the Microcystin molecular engram film 10 that described sensing chip 7 comprises glass sheet 8, is plated in the layer gold 9 of glass sheet 8 single side surface and is modified on layer gold 9 surface, Microcystin molecular engram film 10 has to Microcystin 11 specific recognition and equally distributed micropore 12.
Further, described linear polarization light source be halogen tungsten lamp 1, linear polarizer 2 and the multimode quartz light pricker 3 that is arranged between halogen tungsten lamp 1 and linear polarizer 2; Described spectroanalysis instrument 6 is visible light wave range spectroanalysis instrument, and visible light wave range spectroanalysis instrument forms light-path by multimode quartz light pricker 3 and 45 °/45 °/90 ° glass prisms 4.
Embodiment 1
Thermal polymerization Microcystin molecular engram film is adopted to be modified at layer gold surface: to use oxygen gas plasma washer to clean 2 minutes gold-plated glass sheet, soak more than 12 hours in the ethanolic solution of 50mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.1M and the N-hydroxy-succinamide of 0.01M 1 hour; After taking out directly in immersion 100mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 0.1umol Microcystin, 1umol methacrylic acid and 2umol trimethylol-propane trimethacrylate and add the mixing of 5mL dimethyl sulphoxide solution, nitrogen treatment 10 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 18 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 12 time, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
Embodiment 2
Thermal polymerization Microcystin molecular engram film is adopted to be modified at layer gold surface: to use oxygen gas plasma washer to clean 4 minutes gold-plated glass sheet, soak more than 12 hours in the ethanolic solution of 1000mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 1M and the N-hydroxy-succinamide of 0.1M 1 hour; After taking out directly in immersion 500mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 200umol Microcystin, 2000umol methacrylic acid and 2000umol trimethylol-propane trimethacrylate and add the mixing of 10mL dimethyl sulphoxide solution, nitrogen treatment 12 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 15 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 18 time, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
Embodiment 3
Thermal polymerization Microcystin molecular engram film is adopted to be modified at layer gold surface: to use oxygen gas plasma washer to clean 3 minutes gold-plated glass sheet, soak more than 12 hours in the ethanolic solution of 200mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.2M and the N-hydroxy-succinamide of 0.05M 1 hour; After taking out directly in immersion 200mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 100umol Microcystin, 200umol methacrylic acid and 300umol trimethylol-propane trimethacrylate and add the mixing of 6mL dimethyl sulphoxide solution, nitrogen treatment 13 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 20 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 18 time, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
Embodiment 4
Thermal polymerization Microcystin molecular engram film is adopted to be modified at layer gold surface: to use oxygen gas plasma washer to clean 3 minutes gold-plated glass sheet, soak more than 12 hours in the ethanolic solution of 800mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.8M and the N-hydroxy-succinamide of 0.08M 1 hour; After taking out directly in immersion 400mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 120umol Microcystin, 1500umol methacrylic acid and 1200umol trimethylol-propane trimethacrylate and add the mixing of 8mL dimethyl sulphoxide solution, nitrogen treatment 15 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 16 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 16 time, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
Claims (8)
1. a Microcystin optical detecting sensor, comprise linear polarization light source, 45 °/45 °/90 ° glass prisms, sensing chip, spectroanalysis instruments, it is characterized in that, described sensing chip is provided with silica gel circulation groove, described sensing chip comprises glass sheet, is plated in the layer gold of glass sheet single side surface and is modified at the Microcystin molecular engram film on layer gold surface, Microcystin molecular engram film has to Microcystin specific recognition and equally distributed micropore.
2. a kind of Microcystin optical detecting sensor according to claim 1, is characterized in that, adopts rubbing method that Microcystin molecular engram film is modified at layer gold surface.
3. a kind of Microcystin optical detecting sensor according to claim 1, is characterized in that, adopts thermal polymerization Microcystin molecular engram film to be modified at layer gold surface.
4. a kind of Microcystin optical detecting sensor according to claim 3, it is characterized in that, described hot polymerization is combined into: gold-plated glass sheet is used oxygen gas plasma washer cleaning 2-4 minute, soak more than 12 hours in the ethanolic solution of 50-1000mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.1-1M and the N-hydroxy-succinamide of 0.01-0.1M 1 hour; After taking out directly in immersion 100-500mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 0.1-200umol Microcystin, 1-2000umol methacrylic acid and 2-2000umol trimethylol-propane trimethacrylate and add the mixing of 5-10mL dimethyl sulphoxide solution, nitrogen treatment more than 10 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 18 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 12 time more than, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
5. a kind of Microcystin optical detecting sensor according to claim 1, is characterized in that, the multimode quartz light pricker that described linear polarization light source is halogen tungsten lamp, linear polarizer and is arranged between halogen tungsten lamp and linear polarizer; Described spectroanalysis instrument is visible light wave range spectroanalysis instrument, and visible light wave range spectroanalysis instrument forms light-path by multimode quartz light pricker and 45 °/45 °/90 ° glass prisms.
6. for a sensing chip for Microcystin optical detecting sensor, it is characterized in that, comprise glass sheet, be plated in glass sheet single side surface layer gold, adopt thermal polymerization method to be modified at Microcystin molecular engram film on layer gold surface.
7. a kind of sensing chip for Microcystin optical detecting sensor according to claim 6, is characterized in that, Microcystin molecular engram film has to Microcystin specific recognition and equally distributed micropore.
8. the thermal polymerization method of a kind of sensing chip for Microcystin optical detecting sensor according to claim 6, it is characterized in that, described hot polymerization is combined into: gold-plated glass sheet is used oxygen gas plasma washer cleaning 2-4 minute, soak more than 12 hours in the ethanolic solution of 50-1000mM undecyl mercaptan alkanoic acid, alcohol flushing after taking out, nitrogen dries up; To immerse again in the mixed aqueous solution of 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride of 0.1-1M and the N-hydroxy-succinamide of 0.01-0.1M 1 hour; After taking out directly in immersion 100-500mM amino-butanamide hydrochloride aqueous solution 3 hours, nitrogen dried up; Get 0.1-200umol Microcystin, 1-2000umol methacrylic acid and 2-2000umol trimethylol-propane trimethacrylate and add the mixing of 5-10mL dimethyl sulphoxide solution, nitrogen treatment more than 10 minutes, immerse the glass sheet that pre-service is good, after sealing, under 65 ± 5 DEG C of conditions, carry out heat polymerization 18 hours; Take out after use ethanol and acetic acid equal-volume less than mixed solution flow wash 12 time more than, after completing, alcohol flushing removes surperficial eluent, and nitrogen dries up.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792852A (en) * | 2015-04-09 | 2015-07-22 | 宁波大学 | Algal toxin molecular imprinting chemoreceptor sensor as well as preparation method and application thereof |
| CN105866044A (en) * | 2016-06-23 | 2016-08-17 | 湖北出入境检验检疫局检验检疫技术中心 | Quick detector for microcystic toxins |
| CN106405080A (en) * | 2016-03-07 | 2017-02-15 | 天津科技大学 | Intelligent detection system for microcystins |
| CN106568745A (en) * | 2016-10-27 | 2017-04-19 | 暨南大学 | Microcystic toxin biological chip and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103698242A (en) * | 2013-12-21 | 2014-04-02 | 中国科学院苏州生物医学工程技术研究所 | Rapid detecting sensor for microcystins |
| CN103760052A (en) * | 2013-12-21 | 2014-04-30 | 中国科学院苏州生物医学工程技术研究所 | Piezoelectric detection sensor for microcystic toxins based on molecular imprinting technology |
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- 2014-11-05 CN CN201410617885.XA patent/CN104345037B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103698242A (en) * | 2013-12-21 | 2014-04-02 | 中国科学院苏州生物医学工程技术研究所 | Rapid detecting sensor for microcystins |
| CN103760052A (en) * | 2013-12-21 | 2014-04-30 | 中国科学院苏州生物医学工程技术研究所 | Piezoelectric detection sensor for microcystic toxins based on molecular imprinting technology |
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| HAO HE ET AL: "Detection of trace microcystin-LR on a 20 MHz QCM sensor coated with in situ self-assembled MIPs", 《TALANTA》 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792852A (en) * | 2015-04-09 | 2015-07-22 | 宁波大学 | Algal toxin molecular imprinting chemoreceptor sensor as well as preparation method and application thereof |
| CN104792852B (en) * | 2015-04-09 | 2018-01-16 | 宁波大学 | A kind of Algae toxins molecular engram chemoreceptor sensor and its preparation method and application |
| CN106405080A (en) * | 2016-03-07 | 2017-02-15 | 天津科技大学 | Intelligent detection system for microcystins |
| CN105866044A (en) * | 2016-06-23 | 2016-08-17 | 湖北出入境检验检疫局检验检疫技术中心 | Quick detector for microcystic toxins |
| CN106568745A (en) * | 2016-10-27 | 2017-04-19 | 暨南大学 | Microcystic toxin biological chip and preparation method and application thereof |
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