CN101825629B - Waveguide coupling metal photonic crystal biosensor and detecting method thereof - Google Patents
Waveguide coupling metal photonic crystal biosensor and detecting method thereof Download PDFInfo
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- CN101825629B CN101825629B CN200910079243.8A CN200910079243A CN101825629B CN 101825629 B CN101825629 B CN 101825629B CN 200910079243 A CN200910079243 A CN 200910079243A CN 101825629 B CN101825629 B CN 101825629B
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Abstract
The invention relates to a waveguide coupling metal photonic crystal biosensor and a detecting method thereof, which are used for the high-sensitivity sensation of biomolecule concentration and bioactive molecule idiosyncrasy. The waveguide coupling metal photonic crystal biosensor comprises a base, a waveguide layer and metal photonic crystals arranged on the waveguide layer. When the waveguide coupling metal photonic crystal biosensor is used for detecting, light transmitted by a light source irradiates the metal photonic crystals at an angle, and an optical detector is used for detecting an extinction spectrum of transmitted light passing through the waveguide layer and the base or detecting an extinction spectrum of transmitted light passing through the metal photonic crystals; after receptors are fixed on the metal photonic crystals, a sample solution flows through the surfaces of the metal photonic crystals, the detector is used for detecting the extinction spectrum of the transmitted light which passes through the waveguide layer and the base at the moment or detecting the extinction spectrum of the transmitted light which passes through the metal photonic crystals, and the two extinction spectrums are calculated by secondary extinction spectrums to quantificationally detect ligand concentration. The waveguide coupling metal photonic crystal biosensor has high sensitivity, low cost, simple preparation, simple use method, and the like.
Description
Technical field
The present invention is a kind of Waveguide coupling metal photonic crystal biosensor and detection method thereof, its based on Physical Mechanism be spectroscopy coupling strong between particle plasma resonance in metal photonic crystal and Wave guide resonance pattern, can, for the concentration of biomolecule (protein, carbohydrate molecule, DNA etc.) and the high sensitivity sensing of bioactive molecule idiosyncrasy, belong to the crossing domain of photoelectron technology and biotechnology.
Background technology
The biomolecule sensor of high sensitivity, high-resolution characteristic plays very important effect in the scientific researches such as biology, life science, medical science and practical application.Biology sensor based on spectroscopy response characteristic is because its reliable physics principle and accurate technological means show unique advantage.
The basic structure of traditional surface plasma resonance (SPR) biology sensor as shown in Figure 1.This sensor is by transparency carrier 1, and evaporation is at the metal level 2 of substrate 1 upper surface, prism 6, and light source 7, photodetector 8 forms.Surface plasmon resonance biosensor is such for biospecific reaction sensing principle of work: the light beam from wideband light source 7 incides prism 6 at a certain angle, prism 6 imports transparency carrier by incident light with wide-angle, and the incident light being imported into total reflection occurs and in metal level 2, excite evanescent wave (Evanescent Wave) at the interface of metal level 2 and substrate 1.In given incident angle situation, a certain frequency in evanescent wave reaches consistent by the surface plasma resonance frequency with metal level 2, makes incident light in the strong resonance absorption of this frequency quilt, records the significant absorption peak in one, this frequency place in its reflectance spectrum.Here it is so-called surface plasmon resonance absorption.In actual applications, also can adopt monochromatic source (as laser) as incident light, by changing incident angle, realize the object of metal level to its surface plasmon resonance absorption.The problems such as currently used technology mostly is the former, adopts the mode be fixed into firing angle and wideband light source, and complex operation, the error that can avoid angular adjustment to cause are large, system is unstable.
In bio-sensing experiment, the acceptor 4 that first can react with part 3 generation specific recognition is fixed on metal level 2, because surface plasma resonance frequency depends on the specific inductive capacity (refractive index) of environment consumingly, the metallic film after acceptor molecule is fixing has possessed specific SPR frequency.Then the acceptor and the part in sample solution 53 that are fixed on metallic film 2 are interacted, acceptor and part generation specific reaction, just having there is variation in the specific inductive capacity at layer on surface of metal place, thereby causes the surface plasma resonance frequency of metal level to be offset.Thus, can obtain ligand concentration information in sample solution.
Biology sensor based on SPR technology still has device architecture complexity, technology of preparing and requires high, system complex operation, test period long, thus high in cost of production shortcoming.Therefore, biology sensor simple in structure, highly sensitive, that test process is quick, cost is low becomes an urgent demand and the important research and development content in the fields such as biomedicine.
Summary of the invention
The object of the invention is to overcome the above-mentioned defect of existing biology sensor, proposed a kind of Waveguide coupling metal photonic crystal biosensor and detection method thereof, this biology sensor has the advantages such as highly sensitive, cost is low, preparation and application is simple.
Biology sensor in the present invention based on Physical Mechanism be: strong coupling between particle plasma resonance (PPR) pattern of Wave guide resonance pattern (WGM) and metal photonic crystal, therefore be called " Waveguide coupling metal photonic crystal biosensor ".
The technical solution used in the present invention is as follows: this biology sensor comprises successively from top to bottom substrate 21, covers ducting layer transparent in substrate 21 22, covers the metal photonic crystal 23 on ducting layer 22.
The thickness of described ducting layer 21 is 60nm～300nm.
Described metal photonic crystal 23 is one-dimensional metal photon crystals or two-dimensional metallic photonic crystal.
The cycle of described one-dimensional metal photon crystals is 150nm～550nm.
The cycle of described two-dimensional metallic photonic crystal both direction can be the same or different, and span is 150nm～550nm.
Use above-mentioned Waveguide coupling metal photonic crystal biosensor to detect, its detection method is as follows:
1) on metal photonic crystal 23 after the fixing acceptor 4 part 3 to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends be take and the normal of substrate 21 place planes (scope of θ is 0～80 °) into θ angle irradiation metal photonic crystal, the extinction spectra that photodetector 8 detects through the transmitted light of blank sample solution, metal photonic crystal 23, ducting layer 22 and substrate 21 after Polarization Controller 31;
3) sample solution that contains part is flowed through surface, part and the acceptor generation specific reaction of metal photonic crystal 23.Utilize step 1) in blank sample solution wash after the part not reacting, then by the Surface Contact of blank sample solution and metal photonic crystal 23;
4) light that light source 7 sends becomes and step 2 with the normal with substrate 21 place planes after Polarization Controller 31) identical θ angular illumination metal photonic crystal, the extinction spectra that photodetector 8 detects through the transmitted light of blank sample solution, metal photonic crystal, ducting layer 22 and substrate 21;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
Use above-mentioned Waveguide coupling metal photonic crystal biosensor to detect, also can detect as follows:
1) on metal photonic crystal 23 after the fixing acceptor 4 part 3 to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates substrate with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, the extinction spectra that photodetector 8 detects through the transmitted light of substrate 21, ducting layer 22, metal photonic crystal 23 and blank sample solution;
3) sample solution that contains part is flowed through surface, part and the acceptor generation specific reaction of metal photonic crystal 23.Utilize step 1) blank sample solution wash the part not reacting, finally by blank sample solution again with the Surface Contact of metal photonic crystal 23;
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination substrate, the extinction spectra that photodetector 8 detects through the reacted transmitted light of substrate 21, ducting layer 22, metal photonic crystal 23 and blank sample solution;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
Use above-mentioned Waveguide coupling metal photonic crystal biosensor to detect, also can detect as follows:
1) on metal photonic crystal 23 after the fixing acceptor 4 part 3 to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates metal photonic crystal with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, and photodetector 8 detects the catoptrical extinction spectra being fired back by substrate 21 after blank sample solution, metal photonic crystal and ducting layer 22 successively;
3) sample solution that contains part is flowed through surface, part and the acceptor generation specific reaction of metal photonic crystal 23.Utilize step 1) blank sample solution wash the part not reacting, finally by blank sample solution again with the Surface Contact of metal photonic crystal 23.
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination metal photonic crystal, the catoptrical extinction spectra being reflected by substrate 21 after blank sample solution, metal photonic crystal 23 and ducting layer 22 successively after photodetector 8 detection reaction;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
Use above-mentioned Waveguide coupling metal photonic crystal biosensor to detect, also can detect as follows:
1) on metal photonic crystal 23 after the fixing acceptor 4 part 3 to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates substrate 21 with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, and photodetector 8 detects the catoptrical extinction spectra being reflected by metal photonic crystal after substrate 21 and ducting layer 22 successively;
3) sample solution that contains part is flowed through surface, part and the acceptor generation specific reaction of metal photonic crystal 23.Utilize step 1) blank sample solution wash the part not reacting, finally by the Surface Contact of blank sample solution and metal photonic crystal 23.
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination substrate 21, after photodetector 8 detection reaction successively after substrate 21 and ducting layer 22, the catoptrical extinction spectra being reflected by metal photonic crystal;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
Device in the present invention can also be used to the variation of quantitative detection material concentration, specifically has following several detection method:
One, use the Waveguide coupling metal photonic crystal biosensor in the present invention to carry out the detection of material concentration, can carry out as follows:
1) by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates metal photonic crystal with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, the extinction spectra that photodetector 8 detects through the transmitted light of blank sample solution, metal photonic crystal 23, ducting layer 22 and substrate 21;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal 23, and with the Surface Contact of metal photonic crystal 23.
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination metal photonic crystal, the extinction spectra that photodetector 8 detects through the transmitted light of sample solution, metal photonic crystal 23, ducting layer 22 and substrate 21;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
Two, the difference of this method and method one is to detect light from substrate incident, and concrete steps are as follows:
1) by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates substrate 21 with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, the extinction spectra that photodetector 8 detects through the transmitted light of substrate 21, ducting layer 22, metal photonic crystal 23 and blank sample solution;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal 23, and with the Surface Contact of metal photonic crystal 23.
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination substrate 21, the extinction spectra that photodetector 8 detects through the transmitted light of substrate 21, ducting layer 22, metal photonic crystal 23 and sample solution;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
The method of the detection of reflected light that three, this method adopts is determined the variation of material concentration, and its concrete steps are as follows:
1) by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates metal photonic crystal with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, photodetector 8 detects successively after blank sample solution, metal photonic crystal 23 and ducting layer 22, the catoptrical extinction spectra being reflected by substrate 21;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal 23, and with the Surface Contact of metal photonic crystal 23.
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination metal photonic crystal, photodetector 8 detects successively after sample solution, metal photonic crystal and ducting layer 22, the catoptrical extinction spectra being reflected by substrate 21;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
Four, the difference of this method and method three is to detect light from substrate incident, and concrete steps are as follows:
1) by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) light that light source 7 sends irradiates substrate 21 with the normal degree into θ angle (scope of θ is 0～80 °) of substrate 21 place planes after Polarization Controller 31, and photodetector 8 detects the catoptrical extinction spectra being reflected by metal photonic crystal after substrate 21 and ducting layer 22;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal 23, and with the Surface Contact of metal photonic crystal 23;
4) light that light source 7 sends becomes and step 2 with the normal of substrate 21 place planes after Polarization Controller 31) identical θ angular illumination substrate 21, photodetector 8 detects the catoptrical extinction spectra being reflected by metal photonic crystal after substrate 21 and ducting layer 22 successively;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
The principle of work of this biology sensor is as shown in Figure 5: incident light 24 in metal photonic crystal 23 excited particles plasma resonance excimer Er characteristic spectrum district by strong absorption in, by metal photonic crystal diffraction and in ducting layer 22, excite waveguide propagation modes 27.In waveguide mode communication process, by the metal photonic crystal diffraction of upper surface, produced the diffraction light 28 that multi beam is parallel to transmitted light 26, diffraction light 28 further interferes with transmitted light 26.Like this, in the broadband of the transmitted light recording etc., from the transmission that can observe an arrowband in resonance absorption spectrum, strengthen signal, this is the result of waveguide mode and the effect of plasma resonance Mode Coupling.The particle plasma resonance absorption of metal photonic crystal and the spectroscopy response characteristic of Wave guide resonance pattern are all offset with the variation of metal photonic crystal upper surface environment refractive index, and its side-play amount is directly corresponding to the variable quantity of environment refractive index.Therefore, by measuring the variation of the coupling spectrum of particle plasma resonance absorption and Wave guide resonance pattern, get final product the variable quantity of quantitatively characterizing environment refractive index.The principle of work of biology sensor in the present invention that Here it is.Wherein the introducing of arrowband Wave guide resonance pattern has improved the sensitivity of sensor response greatly.
The detection principle of Waveguide coupling metal photonic crystal biosensor of the present invention is as follows: in biospecific reaction sensing experiment, first the acceptor (antibody) 4 part (antigen) 3 to specific recognition function is fixed on to the surface of metal photonic crystal 23, forms the initial diaphragm of sensor; By the flow through surface of metal photonic crystal 23 of sample solution 5, if there is part (antigen) 3 in test portion, just can there is specific reaction with part (antigen) 3 in acceptor (antibody) 4, according to the variation of extinction spectra before and after reaction, the subtle change of coupling between quantitative waveguide mode and particle plasma resonance pattern, by the quantitatively characterizing to spectrum change rule, realize the detection to the concentration of part (antigen) 3 in sample solution 5.In material concentration test experience, the flow through surface of metal photonic crystal 23 of sample solution 5, if the concentration of test substance changes in test portion, according to the variation of extinction spectra before and after concentration change, the subtle change of coupling between quantitative waveguide mode and particle plasma resonance pattern, by the quantitatively characterizing to spectrum change rule, realize the detection to material concentration to be measured.
The advantageous feature that the present invention is different from traditional spr sensor embodies in the following areas:
1) cost is low: no matter be that Waveguide coupling metal photonic crystal senser element or the cost of spectroscopy test macro are all far below spr sensor system.Whole sensing system cost only has below 10% of existing surface plasmon resonance biosensor price in the market.The Waveguide coupling metal photonic crystal sensor of its core can be reused, and has improved the dirigibility of practical application.
2) highly sensitive: the introducing of Wave guide resonance pattern has realized the narrowband modulation to particle plasma resonance pattern, greatly improved the sensitivity of device to the response of environment variations in refractive index.
3) means of testing is flexible, easy: according to different demands, various test is available; In test process, only need to adopt conventional optics and spectroscopy means of testing, obtain the extinction spectra of transmission or reflectance spectrum or transmission or reflection.
4) be convenient to integrated: Waveguide coupling metal photonic crystal volume is little, and regular shape is easy to embedded system, forms biochip structure.
Accompanying drawing explanation
The surface plasma resonance biological sensor structural representation that Fig. 1 is traditional
The sectional view of Fig. 2 Waveguide coupling metal photonic crystal biosensor of the present invention
The front elevation of Fig. 3 waveguide-coupled one-dimensional metal photon crystals of the present invention biology sensor
The front elevation of Fig. 4 waveguide-coupled two-dimensional metallic photonic crystal of the present invention biology sensor
The optical schematic diagram of Fig. 5 Waveguide coupling metal photonic crystal biosensor of the present invention
The structural drawing of Fig. 6 Waveguide coupling metal photonic crystal biosensor of the present invention
The positive transmission-type of Fig. 7 biology sensor of the present invention detects schematic diagram
The back side transmission-type of Fig. 8 biology sensor of the present invention detects schematic diagram
The reflective detection schematic diagram in front of Fig. 9 biology sensor of the present invention
The backside reflection formula of Figure 10 biology sensor of the present invention detects schematic diagram
The extinction spectra of the transmission-type waveguide-coupled gold photonic crystal biosensor in Figure 11 the present invention
The detection signal of the transmission-type waveguide-coupled gold photonic crystal biosensor in Figure 12 the present invention
The catoptrical extinction spectra of the reflective waveguide-coupled gold photonic crystal biosensor in Figure 13 the present invention
The detection signal of the reflective waveguide-coupled gold photonic crystal biosensor in Figure 14 the present invention
In figure: 1. transparency carrier, 2. metal level, 3. part (antigen), 4. acceptor (antibody), 5. sample solution, 6. prism, 7. light source, 8. photodetector, 21. substrates, 22. ducting layers, 23. metal photonic crystals, 31. Polarization Controllers.
Embodiment
The invention will be further described with reference to the accompanying drawings:
Biology sensor in the present invention comprises substrate 21 (thickness is D), covers the transparent ducting layer 22 (thickness d) in substrate 21 and covers the metal photonic crystal 23 (cycle is Λ) on ducting layer 22, and the sectional view of this biology sensor as shown in Figure 2.Wherein: the span of the thickness d of ducting layer 22 is 60～300nm, metal photonic crystal 23 can adopt one dimension (Fig. 3) or two-dimensional structure (Fig. 4), is called one dimension or two-dimensional metallic photonic crystal.The cycle of one-dimensional metal photon crystals is 150～550nm, and the cycle of 2 D photon crystal both direction all can be taken as 150～550nm.
Biology sensor in the present invention is prepared metal (preferably gold, silver or platinum) one dimension or 2 D photon crystal on the transparent waveguide layer 22 of substrate 21, obtains Waveguide coupling metal photonic crystal biosensor as shown in Figure 2 to 4;
The detection method of the Waveguide coupling metal photonic crystal biosensor in the present invention is as follows:
1) in biologic specificity recognition detection, first the acceptor (antibody) 4 part (antigen) 3 to specific recognition function is fixed on to step 1) in the surface of metal photonic crystal 23 in the Waveguide coupling metal photonic crystal biosensor of preparation, form the initial diaphragm of sensor;
2) spectrum test method can adopt the mode as shown in Fig. 7～Figure 10:
3) surface of biology sensor sample solution 5 is flowed through step 3) obtaining, if there is part (antigen) 3 in test portion, acceptor (antibody) 4 just with its generation specific reaction, according to the variation of extinction spectra before and after reaction, the subtle change (as shown in Figure 11 or Figure 13) of coupling between quantitative waveguide mode and particle plasma resonance pattern, by characterizing the spectroscopy Changing Pattern of variable concentrations antigen, realize the quantitative detection to part (antigen) 3 concentration in sample solution 5.
The detection of the Waveguide coupling metal photonic crystal biosensor in the present invention also can be adopted with the following method:
1) Surface Contact with metal photonic crystal 23 by blank sample solution (solvent);
2) spectrum test method can adopt the mode as shown in Fig. 7～Figure 10;
3) Surface Contact with metal photonic crystal 23 by the sample solution that contains certain material (solution of test substance), according to the variation of sample solution concentration, the subtle change (as shown in Figure 11 or Figure 13) of coupling between quantitative waveguide mode and particle plasma resonance pattern, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration;
Be the device in the present invention and the example of detection method thereof below:
Embodiment 1:
The present embodiment is positive transmission-type waveguide-coupled one-dimensional metal photon crystals biology sensor and detection method thereof:
The substrate 21 of using in the present embodiment adopts the glass of thickness D=1mm, and ducting layer 22 adopts tin indium oxide (ITO) film, the glass sheet that thickness is d=200nm.
It is Λ=330nm one dimension gold photonic crystal that the present embodiment adopts preparation method's manufacturing cycle on the ITO of substrate of glass ducting layer of interference lithography binding soln method, obtains waveguide-coupled one dimension gold photonic crystal biosensor.While utilizing this sensor to detect, its detection method is as follows:
1) acceptor (antibody) 4 part (antigen) 3 to specific recognition function is fixed on to the surface of the metal photonic crystal 23 in waveguide-coupled one-dimensional metal photon crystals biology sensor, form the initial diaphragm of sensor, the Surface Contact by blank sample solution (buffer solution or the solvent that do not contain test substance) with metal photonic crystal 23;
2) spectrum test mode adopts positive transmission-type as shown in Figure 7, light source is selected bromine tungsten filament lamp, incident light becomes the waveguide-coupled one-dimensional metal photon crystals in ° angle the present embodiment of θ=32 to irradiate light through Polarization Controller 31 with the normal with substrate 21 place planes, the extinction spectra that photodetector 8 detects through the transmitted light of blank sample solution, one-dimensional metal photon crystals, ducting layer and substrate;
3) by the sample solution that contains part (antigen) 3 (buffer solution that contains test substance or the solvent) step 2 of flowing through) surface of the biology sensor that obtains, there is specific recognition with acceptor (antibody) 4 and react in part (antigen) 3, utilize step 2) blank sample solution wash the antigen not reacting, finally by the Surface Contact of blank sample solution and metal photonic crystal 23;
4) light that bromine tungsten filament lamp light source sends is after Polarization Controller 31, with the normal with substrate 21 place planes, become θ=32 ° angular illumination one-dimensional metal photon crystals, photodetector detects the extinction spectra of transmitted light after the reaction of blank sample solution, one-dimensional metal photon crystals, ducting layer and substrate;
5) step 2) with step 4) middle extinction spectra comparison, the subtle change of coupling between waveguide mode and particle plasma resonance pattern before and after acquisition reaction as shown in figure 11, extinction spectra is made to secondary extinction spectral manipulation, obtain signal as shown in figure 12, by characterizing the spectroscopy Changing Pattern of different antigen concentrations, realize the quantitative detection to antigen concentration.
Embodiment 2:
The present embodiment is back side transmission-type two-dimensional waveguide coupling metal photonic crystal biosensor and device detection method:
The substrate of using in the present embodiment adopts the glass of thickness D=1mm, and ducting layer adopts ITO, the glass sheet that thickness is d=200nm.
The present embodiment adopts the preparation method of interference lithography binding soln method on the ITO of substrate of glass ducting layer, to prepare the both direction cycle and is Λ=350nm two-dimensional gold photonic crystal, obtains waveguide-coupled two-dimensional gold photonic crystal biosensor.While utilizing this sensor to detect, its detection method is as follows:
1) acceptor (antibody) 4 part (antigen) 3 to specific recognition function is fixed on to the surface of metal photonic crystal in two-dimensional waveguide coupling metal photonic crystal biosensor, form the initial diaphragm of sensor, by the Surface Contact of blank sample solution and metal photonic crystal;
2) spectrum test mode adopts the back side (basal surface) transmission-type as shown in Figure 8, light source is selected bromine tungsten filament lamp, incident light becomes the substrate of the waveguide-coupled two-dimensional metallic photonic crystal biology sensor in ° angle the present embodiment of θ=48 to irradiate with the normal with substrate 21 place planes after Polarization Controller, and photodetector detects the extinction spectra through the transmitted light of substrate, ducting layer, two-dimensional metallic photonic crystal and blank sample solution;
3) surface of the biology sensor sample solution that contains antigen is flowed through step 2) obtaining, antigen reacts with antibody generation specific recognition, utilize step 2) blank sample solution wash the antigen not reacting, finally by the Surface Contact of blank sample solution and metal photonic crystal 23.
4) light that bromine tungsten filament lamp light source sends is after Polarization Controller, with the normal with substrate 21 place planes, become θ=48 ° angular illumination substrate, photodetector detects incident light through the extinction spectra of the transmitted light of substrate, ducting layer, two-dimensional metallic photonic crystal and blank sample solution;
5) step 2) with step 4) middle extinction spectra comparison, obtain the subtle change of coupling between waveguide mode as shown in figure 11 and particle plasma resonance pattern, extinction spectra is made to secondary extinction spectral manipulation, obtain signal as shown in figure 12, by characterizing the spectroscopy Changing Pattern of different antigen concentrations, realize the quantitative detection to antigen concentration.
Embodiment 3:
The present embodiment is positive reflective waveguide-coupled one-dimensional metal photon crystals biology sensor and detection method thereof:
The substrate of using in the present embodiment adopts the glass of thickness D=1mm, and ducting layer adopts ITO, the glass sheet that thickness is d=210nm.
It is Λ=400nm one dimension gold photonic crystal that the present embodiment adopts preparation method's manufacturing cycle in the ITO of substrate of glass waveguide of interference lithography binding soln method, obtains waveguide-coupled one dimension gold photonic crystal biosensor;
1) antibody antigen to specific recognition function is fixed on to the surface of metal photonic crystal in waveguide-coupled one-dimensional metal photon crystals biology sensor, form the initial diaphragm of sensor, by the Surface Contact of blank sample solution and metal photonic crystal 23;
2) it is reflective that spectrum test mode adopts front as shown in Figure 9, light source is selected bromine tungsten filament lamp, incident light becomes the one-dimensional metal photon crystals face of the waveguide-coupled one-dimensional metal photon crystals biology sensor in ° angle the present embodiment of θ=2 to irradiate with the normal with substrate 21 place planes after Polarization Controller, irradiate light and after blank sample solution, one-dimensional metal photon crystals, ducting layer, by substrate 21, reflected successively, photodetector 8 receives this catoptrical extinction spectra;
3) surface of the biology sensor sample solution that contains antigen is flowed through step 2) obtaining, antigen reacts with antibody generation specific recognition, utilize step 2) blank sample solution wash the antigen not reacting, finally by the Surface Contact of blank sample solution and metal photonic crystal 23.
4) light that bromine tungsten filament lamp light source sends is after Polarization Controller, with the normal with substrate 21 place planes, become θ=2 ° angular illumination one-dimensional metal photon crystals, photodetector detects after blank sample solution, one-dimensional metal photon crystals, ducting layer, the reacted catoptrical extinction spectra being reflected by substrate;
5) step 2) with step 4) middle extinction spectra comparison, the subtle change of coupling between waveguide mode and particle plasma resonance pattern before and after acquisition reaction as shown in figure 13, extinction spectra is made to secondary extinction spectral manipulation, obtain signal as shown in figure 14, by characterizing the spectroscopy Changing Pattern of different antigen concentrations, realize the quantitative detection to antigen concentration.
Embodiment 4:
The present embodiment is backside reflection formula one-dimensional wave guide coupling metal photonic crystal biosensor and detection method thereof: the device in the present embodiment is identical with embodiment 3, its detection method is substantially the same manner as Example 3, difference is only: incident light is after Polarization Controller, incide in substrate 21, what detecting device 8 detected is that incident light is after substrate, ducting layer, metal photonic crystal, by the catoptrical extinction spectra of metal photonic crystal.
Sensor in the present invention also can be realized the detection to material concentration, below in conjunction with concrete detection method, describes:
Embodiment 5:
The present embodiment is positive transmission-type two-dimensional waveguide coupling metal photonic crystal biosensor and device detection method:
The substrate of using in the present embodiment adopts the glass of thickness D=1mm, and ducting layer adopts ITO, the glass sheet that thickness is d=200nm.
The present embodiment adopts the preparation method of interference lithography binding soln method on the ITO of substrate of glass ducting layer, to prepare the both direction cycle and is Λ=350nm two-dimensional gold photonic crystal, obtains waveguide-coupled two-dimensional gold photonic crystal biosensor.The detection of device in use the present embodiment to material concentration, the step of its detection is as follows:
1) Surface Contact with two-dimensional metallic photonic crystal 23 by blank test portion (pure water);
2) light that light source 7 sends becomes 36 ° of angles to irradiate metal photonic crystal after Polarization Controller 31 with the normal of substrate 21 place planes, photodetector 8 detects the extinction spectra of the transmitted light that passes through successively pure water, metal photonic crystal 23 ducting layers 22 and substrate 21;
3) by the sample solution stream (aqueous solution of sucrose) that contains sucrose through the surface of metal photonic crystal 23, and with the Surface Contact of metal photonic crystal 23;
4) light that light source 7 sends becomes 36 ° of angles to irradiate metal photonic crystal after Polarization Controller 31 with the normal of substrate 21 place planes, photodetector 8 detects the extinction spectra of the transmitted light that passes through successively sucrose solution, metal photonic crystal 23, ducting layer 22 and substrate 21;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, obtain as shown in figure 14 because sucrose concentration changes the signal producing, by characterizing the spectroscopy Changing Pattern of variable concentrations sucrose, realize the quantitative detection to sucrose concentration.
Embodiment 6:
The present embodiment is back side transmission-type one-dimensional wave guide coupling metal photonic crystal biosensor and detection method thereof: the device in the present embodiment is identical with embodiment 5, its detection method is substantially the same manner as Example 5, difference is only: incident light is after Polarization Controller, incide in substrate 21, what photodetector 8 detected is the extinction spectra of incident light transmitted light after substrate, ducting layer, metal photonic crystal.
Embodiment 7:
The present embodiment is positive reflective one-dimensional wave guide coupling metal photonic crystal biosensor and detection method thereof: the device in the present embodiment is identical with embodiment 5, its detection method is substantially the same manner as Example 5, difference is only: incident light is after Polarization Controller, incide on metal photonic crystal 23, detecting device 8 detects be incident light after metal photonic crystal, ducting layer, substrate, the catoptrical extinction spectra being reflected by substrate 21.
Embodiment 8:
The present embodiment is backside reflection formula one-dimensional wave guide coupling metal photonic crystal biosensor and detection method thereof: the device in the present embodiment is identical with embodiment 5, its detection method is substantially the same manner as Example 5, difference is only: incident light is after Polarization Controller, incide in substrate 21, photodetector 8 detects be incident light successively after substrate, ducting layer, metal photonic crystal, the catoptrical extinction spectra being reflected by metal photonic crystal 21.
Claims (7)
1. Waveguide coupling metal photonic crystal biosensor, is characterized in that: comprise successively from top to bottom substrate, cover suprabasil transparent ducting layer and cover the metal photonic crystal on ducting layer;
Described substrate adopts the glass that thickness is 1mm; Described ducting layer adopts tin indium oxide ito thin film, and thickness is 200nm; The preparation method's manufacturing cycle on the ITO of substrate of glass ducting layer that adopts interference lithography binding soln method is 330nm one dimension gold photonic crystal, obtains waveguide-coupled one dimension gold photonic crystal biosensor;
Or described substrate adopts the glass that thickness is 1mm; Described ducting layer adopts tin indium oxide ito thin film, and thickness is 200nm; Adopt the preparation method of interference lithography binding soln method on the ITO of substrate of glass ducting layer, to prepare the both direction cycle and be 350nm two-dimensional gold photonic crystal, obtain waveguide-coupled two-dimensional gold photonic crystal biosensor;
Or described substrate adopts the glass that thickness is 1mm; Described ducting layer adopts tin indium oxide ito thin film, and thickness is 210nm; The preparation method's manufacturing cycle in the ITO of substrate of glass waveguide that adopts interference lithography binding soln method is 400nm one dimension gold photonic crystal, obtains waveguide-coupled one dimension gold photonic crystal biosensor;
2. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) on metal photonic crystal after fixing acceptor part to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to be radiated on metal photonic crystal with the normal with substrate place plane after Polarization Controller, and photodetector detects the extinction spectra through the transmitted light of blank sample solution, metal photonic crystal, ducting layer and substrate;
3) sample solution that contains part the is flowed through surface of metal photonic crystal, part and acceptor generation specific reaction; Utilize step 1) in blank sample solution wash after the part not reacting, then by the Surface Contact of blank sample solution and metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal with substrate place plane after Polarization Controller) identical angular illumination metal photonic crystal, photodetector detects the extinction spectra through the transmitted light of blank sample solution, metal photonic crystal, ducting layer and substrate;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to part.
3. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) on metal photonic crystal after fixing acceptor part to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to irradiate substrate after Polarization Controller with the normal of substrate place plane, photodetector detects the extinction spectra through the transmitted light of substrate, ducting layer, metal photonic crystal and blank sample solution;
3) sample solution that contains part the is flowed through surface of metal photonic crystal, part and acceptor generation specific reaction, utilize step 1) blank sample solution wash the part not reacting, finally by blank sample solution again with the Surface Contact of metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal of substrate place plane after Polarization Controller) identical angular illumination substrate, photodetector detects the extinction spectra through the reacted transmitted light of substrate, ducting layer, metal photonic crystal and blank sample solution;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
4. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) on metal photonic crystal after fixing acceptor part to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to irradiate metal photonic crystal after Polarization Controller with the normal of substrate place plane, photodetector detects the catoptrical extinction spectra being fired back by substrate after blank sample solution, metal photonic crystal and ducting layer successively;
3) sample solution that contains part the is flowed through surface of metal photonic crystal, part and acceptor generation specific reaction, utilize step 1) blank sample solution wash the part not reacting, finally by blank sample solution again with the Surface Contact of metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal of substrate place plane after Polarization Controller) identical angular illumination metal photonic crystal, the catoptrical extinction spectra being reflected by substrate after blank sample solution, metal photonic crystal and ducting layer successively after photodetector detection reaction;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
5. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) on metal photonic crystal after fixing acceptor part to specific recognition function by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to irradiate substrate after Polarization Controller with the normal of substrate place plane, photodetector detects the catoptrical extinction spectra being reflected by metal photonic crystal after substrate and ducting layer successively;
3) sample solution that contains part the is flowed through surface of metal photonic crystal, part and acceptor generation specific reaction, utilize step 1) blank sample solution wash the part not reacting, finally by the Surface Contact of blank sample solution and metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal of substrate place plane after Polarization Controller) identical angular illumination substrate, after photodetector detection reaction successively after substrate and ducting layer, the catoptrical extinction spectra being reflected by metal photonic crystal;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of different ligands concentration, realize the quantitative detection to ligand concentration.
6. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to irradiate metal photonic crystal after Polarization Controller with the normal of substrate place plane, photodetector detects the extinction spectra of the transmitted light that passes through successively blank sample solution, metal photonic crystal, ducting layer and substrate, or irradiation substrate, photodetector detects the extinction spectra of the transmitted light that passes through successively substrate, ducting layer, metal photonic crystal and blank sample solution;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal, and with the Surface Contact of metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal of substrate place plane after Polarization Controller) identical angular illumination metal photonic crystal, photodetector detects the extinction spectra of the transmitted light that passes through successively sample solution, metal photonic crystal, ducting layer and substrate, or irradiation substrate, photodetector detects the extinction spectra of the transmitted light that passes through successively substrate, ducting layer, metal photonic crystal and blank sample solution;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
7. right to use requires the method that the Waveguide coupling metal photonic crystal biosensor described in 1 carries out the diagnosis of non-disease and the detection of methods for the treatment of, it is characterized in that, the method comprises the following steps:
1) by the Surface Contact of blank sample solution and metal photonic crystal;
2) light that light source sends becomes 0～80 ° of angle to irradiate metal photonic crystal after Polarization Controller with the normal of substrate place plane, photodetector detects after blank sample solution, metal photonic crystal and ducting layer, the catoptrical extinction spectra being reflected by substrate, or irradiation substrate, photodetector detects successively after substrate, ducting layer, the catoptrical extinction spectra being reflected by metal photonic crystal;
3) sample solution that contains certain material the is flowed through surface of metal photonic crystal, and with the Surface Contact of metal photonic crystal;
4) light that light source sends becomes and step 2 with the normal of substrate place plane after Polarization Controller) identical angular illumination metal photonic crystal, photodetector detects after blank sample solution, metal photonic crystal and ducting layer, the catoptrical extinction spectra being reflected by substrate, or irradiation substrate, photodetector detects successively after substrate, ducting layer, the catoptrical extinction spectra being reflected by metal photonic crystal;
5) by step 2) with step 4) in extinction spectra make secondary extinction spectrum and calculate, by characterizing the spectroscopy Changing Pattern of this material variable concentrations, realize the quantitative detection to this material concentration.
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| CN108508204A (en) * | 2018-01-31 | 2018-09-07 | 财团法人交大思源基金会 | Photonic crystal and magnetic bead combined fluorescence immunoassay device and method |
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| CN103454253B (en) * | 2013-06-25 | 2016-04-06 | 复旦大学 | Based on the organic phosphorus detection method of surface plasma body resonant vibration |
| CN103969185B (en) * | 2014-05-19 | 2016-09-14 | 上海理工大学 | A kind of method improving guide mode resonance biosensor test sensitivity |
| US9702813B2 (en) | 2014-07-23 | 2017-07-11 | Infineon Technologies Ag | Sensing systems and methods using a coupling structure |
| EP3415917A4 (en) * | 2016-02-09 | 2019-09-11 | Sekisui Chemical Co., Ltd. | Implement for inspection, inspecting device and inspecting method |
| CN110488510A (en) * | 2019-08-22 | 2019-11-22 | 中北大学 | A kind of polarization regulation super structure surface optical device based on the nearly null effect of ITO dielectric constant |
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| CN108508204A (en) * | 2018-01-31 | 2018-09-07 | 财团法人交大思源基金会 | Photonic crystal and magnetic bead combined fluorescence immunoassay device and method |
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