CN102590147B - Surface plasma resonance system capable of adjusting sensitivity and dynamic range and method thereof - Google Patents
Surface plasma resonance system capable of adjusting sensitivity and dynamic range and method thereof Download PDFInfo
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- CN102590147B CN102590147B CN201210022196.5A CN201210022196A CN102590147B CN 102590147 B CN102590147 B CN 102590147B CN 201210022196 A CN201210022196 A CN 201210022196A CN 102590147 B CN102590147 B CN 102590147B
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Abstract
The invention discloses a surface plasma resonance system capable of adjusting sensitivity and dynamic range and a method thereof. The surface plasma resonance system consists of a reflective index adjustable surface plasma resonance coupling unit, a wide waveband polychromatic light source, an incident beam shaping unit and a reflected light detection unit, wherein the reflective index adjustable surface plasma resonance coupling unit comprises a reflective index adjustable fluid prism, a sensing chip and a sample flowing cell; the sensing chip is positioned on the fluid prism; the sample flowing cell is positioned on the sensing chip; the sensing chip comprises an optical transparent substrate and a sensing membrane positioned on the optical transparent substrate; the optical transparent substrate is contacted with a fluid medium in the fluid prism; and the sensing membrane forms one bottom surface of the sample flowing cell. The system can perform high-sensitivity detection analysis by aiming at various samples to be detected on the premise of guaranteeing the dynamic range. The system is not provided with a mechanical moving part and has high stability.
Description
Technical field
The invention belongs to testing tool technical field, be specifically related to a kind of sensitivity and dynamic range adjustable surface plasma body resonant vibration system and detection method.
background technology
Surface plasma body resonant vibration, as a kind of mark, real-time interface analysis technology of original position exempted from, has been widely used in the fields such as chemistry, biology, medical science, physics, material.To be condition that surface plasma body resonant vibration occurs on the interface based between sensing membrane and dielectric sample change with the variation of sample refractive index on interface the ultimate principle of Applications of surface plasmon resonance.Can pass through the detection of effects on surface plasma resonance condition thus, obtain refractive index and the thickness information of sensing membrane surface sample; And the detection changing by effects on surface plasma resonance condition, can the physics on sensing membrane surface and chemical reaction be monitored and be analyzed, if target molecule is bonded in to sensing membrane surface, in analyte specific biomolecule will with target molecule generation specific binding, can carry out interaction of molecules analysis.
Based on Applications of surface plasmon resonance, surface plasma body resonant vibration and surface plasma resonance image-forming two alanysis systems have been developed.High sensitivity is almost pursuing a goal of all analytic systems, and surface plasma body resonant vibration is no exception.At present, in the practical application of surface plasma body resonant vibration and imaging, developed the raising sensitivity method based on sample is modified and sensing membrane is modified, but these methods limitation method that to be all specific aims very strong, and improve sensitivity and also can bring reducing of dynamic range.If thereby can set up a kind of sensitivity and dynamic range adjustable, the surface plasma resonance sensing method of tool universality is extremely important and significant.
summary of the invention
The object of the present invention is to provide a kind of have universality, sensitivity and the adjustable surface plasma body resonant vibration system of dynamic range.This system can, for various testing samples, guarantee, under the prerequisite of dynamic range, to carry out high-sensitivity detection analysis; And this system machinery-free moving-member, system stability is high.
The surface plasma body resonant vibration system that sensitivity provided by the present invention and dynamic range are adjustable, is comprised of the adjustable surface plasma body resonant vibration coupling unit of refractive index, broadband polychromatic source, incident beam shaping unit and reflected light detecting unit; Wherein, the adjustable surface plasma body resonant vibration coupling unit of described refractive index comprises the adjustable fluid prism of a refractive index, is positioned at the sensing chip on described fluid prism and is positioned at the sample flow cell on described sensing chip; Described sensing chip comprises an optical clear substrate and the sensing membrane being located thereon, and described optical clear substrate contacts with the fluid media (medium) in described fluid prism, and described sensing membrane forms a bottom surface of described sample flow cell.
The light shaping that described incident beam shaping unit sends broadband polychromatic source is the adjustable p polarization parallel light of bulk or the little angle of p polarization converging beam, through fluid prism, is incident in sensing chip; Described reflected light detecting unit is for resolving the catoptrical spectrum through fluid prism.
In the present invention, the fluid prism in the surface plasma body resonant vibration coupling unit that described refractive index is adjustable is comprised of cavity and the fluid media (medium) that is positioned at chamber; The cavity of wherein said fluid prism is provided with two apertures and a macropore, and two apertures are used separately as input and the delivery outlet of described fluid media (medium), and macropore is as the installing port of described sensing chip, so that sensing chip directly contacts with fluid media (medium).
The cavity of described fluid prism can be half round post, semisphere, triangle etc., or other adjustable shape etc.This cavity can be made by optically transparent glass, quartz, silicon, plastic or other material.
Fluid media (medium) in described fluid prism can be various pure or the liquid, the gas that mix such as optically transparent water, oil, ionic liquid, magnetic fluid, or supercritical fluid etc.; The refractive index of described fluid prism can be carried out continuous or discontinuous adjusting by displacement fluid medium or by pressure, temperature, electric field, magnetic field, electromagnetic wave etc.
Described sensing chip comprises an optical clear substrate and preparation sensing membrane thereon, can to sensing membrane, carry out functionalized modification as required; Described optical clear substrate can be glass, quartz, silicon, plastics etc.; Described sensing membrane is the material that surface plasma body resonant vibration can occur gold, silver etc.
Optical clear substrate in described sensing chip directly contacts with the fluid media (medium) in described fluid prism, and the sensing membrane in described sensing chip directly contacts with sample.
Described incident beam shaping unit is comprised of optical alignment element, linear polarizer, adjustable diaphragm, lens etc.
Described reflected light detecting unit is by signal collection element and spectrum resolution installation composition; Described spectrum resolution device can be that monochromator coordinates the photo-electric conversion elements such as photodiode, photoelectric cell, photomultiplier or spectrograph to coordinate the linear array photo-electric conversion elements such as photodiode, charge-coupled image sensor (CCD); Described signal collection element and spectrum resolution device can direct-couplings or are passed through coupling fiber.
The light shaping that described incident beam shaping unit sends broadband polychromatic source is the adjustable p polarization parallel light of bulk or little angle converging beam, optical clear substrate through fluid prism, sensing chip, focus on optical clear substrate and sensing membrane interface and suppressed attenuated total internal reflection occurs, reflected light is through optical clear substrate, the fluid prism of sensing chip, by reflected light detecting unit, carry out spectrum resolution, obtain wavelength resonances surface plasma body resonant vibration curve.
The refractive index of described fluid prism can occur under the condition of surface plasma body resonant vibration, and refractive index is lower, and detection sensitivity is higher, and dynamic range is narrower; And refractive index is higher, detection sensitivity is lower, and dynamic range is wider.
Another object of the present invention is to provide a kind of method of utilizing above-mentioned sensitivity and the adjustable surface plasma body resonant vibration system of dynamic range to detect testing sample.
The method comprises the steps: 1) according to the refractive index of testing sample, select to realize the refractive index of the fluid prism of surface plasma body resonant vibration; 2) testing sample is joined in the sample flow cell of the surface plasma body resonant vibration system described in any one in claim 1-7; 3) utilize the surface plasma body resonant vibration system described in any one in claim 1-7 to detect, obtain wavelength resonances surface plasma body resonant vibration curve.
According to the refractive index of testing sample (given value or estimated value), selection can realize the minimum fluid Refractive Index of Glass Prism of surface plasma body resonant vibration, now can obtain maximum sensitivity; According to the maximum refractive index of testing sample group (given value or estimated value), selection can realize the minimum fluid Refractive Index of Glass Prism of surface plasma body resonant vibration, now can guarantee, under the prerequisite of dynamic range, to obtain maximum sensitivity; For unknown sample or be difficult to the transactional analysis estimated, can select suitable fluid Refractive Index of Glass Prism by preliminary experiment.
The surface plasma body resonant vibration system that sensitivity provided by the present invention and dynamic range are adjustable, based on flexible, the continuous or discontinuous adjustable fluid prism of a refractive index, it can, for various testing samples, guarantee, under the prerequisite of dynamic range, to carry out high-sensitivity detection analysis.Machinery-free moving-member in surface plasma body resonant vibration system of the present invention, system stability is high, can be used for the analysis of various samples, the sample analysis that is particularly useful for sensitivity to have relatively high expectations.The present invention is equally applicable to sensitivity and the dynamic range adjustment of surface plasma resonance image-forming.
Accompanying drawing explanation
Fig. 1 is the general structure schematic diagram of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range.
Fig. 2 is the vertical view of fluid prism in the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range.
Fig. 3 is the light path schematic diagram of the incident beam shaping unit of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range.
Fig. 4 is the light path schematic diagram of the reflected light detecting unit of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range.
Fig. 5 is for adopting the experimental analysis figure that the fluid Refractive Index of Glass Prism of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range is 1.4502.
Fig. 6 is for adopting the experimental analysis figure that the fluid Refractive Index of Glass Prism of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range is 1.5174.
Fig. 7 is for adopting the sample segment enlarged drawing of the experimental analysis figure that the fluid Refractive Index of Glass Prism of the adjustable surface plasma body resonant vibration system of sensitivity of the present invention and dynamic range is 1.5174.
Embodiment
Below in conjunction with accompanying drawing and exemplary embodiments, describe the present invention.Following examples are only for explaining the present invention, but are not used for limiting the scope of the invention.
The general structure schematic diagram of surface plasma body resonant vibration system provided by the invention as shown in Figure 1.It comprises broadband polychromatic source and incident beam shaping unit 1, surface plasma body resonant vibration coupling unit 2 and reflected light detecting unit 3 that refractive index is adjustable.
Wherein, the adjustable surface plasma body resonant vibration coupling unit 2 of described refractive index comprises the adjustable fluid prism 4 of a refractive index, is positioned at sensing chip 6 on fluid 5, is positioned at the sample flow cell 7 on sensing chip 6; Described broadband polychromatic source and the incident beam shaping unit 1 output little angle of broadband polychrome p polarization converging beam 8, described converging beam 8 is incident in sensing chip 6 through fluid prism 4; Described reflected light detecting unit 3 is for resolving the spectrum through the reflected light 9 of fluid prism 4.
Described fluid prism 4 has half-cylindrical cavity 10 and is contained in the fluid media (medium) 5 in cavity.On the cavity 10 of half-cylindrical fluid prism, have 11,12 and macropores 13 of two apertures (as shown in Figure 1, 2), two apertures 11,12 are used separately as input port and the delivery outlet of described fluid media (medium), and macropore 13 is for the mounting hole of sensing chip 6; The refractive index of described fluid prism 4 can regulate by changing fluid media (medium) 5.In the present embodiment, the cavity of fluid prism 4 is made by K9 glass material, and fluid media (medium) 5 is the mixed solution of cedar oil and isopropyl alcohol, can regulate continuously by the ratio of cedar oil and isopropyl alcohol in adjusting mixed solution the refractive index of fluid prism 4.
Described sensing chip 6 comprises an optical clear substrate 14 and preparation sensing membrane 15 thereon.In the present embodiment, optical clear substrate 14 is the K9 glass sheet of a thickness 0.15mm, and sensing membrane is the golden film of thickness 50nm.Sensing chip 6 is realized sealing at macropore 13 places and the fluid prism 4 of fluid prism 4 by pressure by dimethyl silicone polymer (PDMS) elastomeric pad 16.The optical clear substrate 14 of sensing chip 6 directly contacts with the fluid media (medium) 5 in fluid prism 4, and sensing membrane 15 directly contacts with sample 17.
Described sample flow cell 7 is that a K9 glass sheet 18 is realized sealing by PDMS pad 19 and sensing chip 6 by pressure; On K9 glass sheet 18, have two apertures 20,21, as the sample of sample flow cell, input and delivery outlet respectively.
Described broadband polychromatic source and incident beam shaping unit 1 as shown in Figure 3, are comprised of broadband polychromatic source 22, optical alignment element 23, linear polarizer 24, adjustable diaphragm 25, long burnt optical lens 26.In the present embodiment, broadband polychromatic source 22 is an iodine-tungsten lamp, and the light sending is collimated by optical lens 30 after two optical lenses 27,28 focus on, and for improving collimation effect, in focusing, an aperture 29 is placed at focus place; Collimation directional light through linear polarizer 24, adjustable diaphragm 25, long burnt optical lens 26, finally obtains the little angle of p polarization converging beam 8 again.Converging beam 8 is through 4 incidents of fluid prism and focus on the optical clear substrate 14 of sensing chip 6 and the interface of sensing membrane 15.
Described reflected light detecting unit 3 is comprised of as signal collection element and spectrum resolution device 33 optical lens 31; In the present embodiment, reflected light 9 focuses on and is coupled in optical fiber 32 through optical collection lens 31, by spectrum resolution device 33 independently, carries out spectrum resolution.Spectrum resolution device 33 is detected by photomultiplier for after monochromator splitting.
The little angle of the broadband polychrome p polarization converging beam 8 of described broadband polychromatic source and 1 output of incident beam shaping unit, optical clear substrate 14 through fluid prism 4, sensing chip, at optical clear substrate 14 and the interface of sensing membrane 15, there is suppressed attenuated total internal reflection, reflected light 9 is through optical clear substrate 14, the fluid prism 4 of sensing chip, by reflected light detecting unit 3, carry out spectrum resolution, obtain wavelength resonances surface plasma body resonant vibration curve.In the present embodiment, the incident direction of incident light is fixed, and between the normal of sensing chip, angle is about 73 degree.
Adopt the adjustable surface plasma body resonant vibration system of a kind of sensitivity that the present embodiment sets up and dynamic range, the aqueous sucrose solution of various concentration of take is sample, has carried out experimental analysis.
First sucrose concentration is respectively to 0% (refractive index n
0%=1.3330), 1% (refractive index n
1%=1.3342) 2 samples, selecting refractive index is that 1.4502 cedar oil aqueous isopropanol (mass ratio of cedar oil and isopropyl alcohol is 1.28: 1) is as the fluid media (medium) of fluid prism, carried out high-sensitivity surface plasma resonance analysis (as shown in Figure 5), corresponding surface plasma body resonant vibration wavelength is respectively λ
0%=776.60nm, λ
1%=792.30nm, calculating wavelength sensitivity is (λ
1%-λ
0%)/(n
1%-n
0%)=13083.3nm/RIU; Then the fluid media (medium) of fluid prism is changed to refractive index and is 1.5174 cedar oil, sucrose concentration is respectively to 0% (refractive index n
0%=1.3330), 1% (refractive index n
1%=1.3342), 5% (refractive index n
5%=1.3401), 13% (refractive index n
13%=1.3529), 18% (refractive index n
18%=1.3605) 5 samples have carried out surface plasma body resonant vibration analysis (as shown in Figure 6,7), and corresponding surface plasma body resonant vibration wavelength is respectively λ
0%=593.44nm, λ
1%=595.62nm, λ
5%=603.03nm, λ
13%=623.48nm, λ
18%=637.58nm, it is 1816.7nm/RIU that 2 samples by 0% and 1% calculate wavelength sensitivity.By analysis result, can be found out, the refractive index of fluid prism is that the sensitivity of 1.4502 o'clock is that the refractive index of fluid prism is 1.5174 o'clock 7 times, and dynamic range narrows down.Thus, only, by regulating the refractive index of the fluid media (medium) of fluid prism, can carry out for testing sample the optimization of sensitivity and dynamic range.Because the refractive index of fluid media (medium) can regulate continuously by the concentration ratio of mixed solution, so can guarantee, under the condition that dynamic range is met, to carry out high-sensitivity detection analysis.
Described embodiment has adopted half-cylindrical fluid prism, also can adopt other shapes such as triangle.
Described embodiment has adopted monochromator to separate analysis of spectrum by coupling fiber, also can adopt spectrograph through spectrum unscrambling analytical equipments such as CCD, also can be without coupling fiber Direct Analysis.
Described embodiment has set up a kind of sensitivity and the adjustable surface plasma body resonant vibration system of dynamic range, and the present invention is equally applicable to sensitivity and the dynamic range adjustment of surface plasma resonance image-forming.
Claims (7)
1. the adjustable surface plasma body resonant vibration system of sensitivity and dynamic range, is comprised of the adjustable surface plasma body resonant vibration coupling unit of refractive index, broadband polychromatic source, incident beam shaping unit and reflected light detecting unit; Wherein, the adjustable surface plasma body resonant vibration coupling unit of described refractive index comprises the adjustable fluid prism of a refractive index, is positioned at the sensing chip on described fluid prism and is positioned at the sample flow cell on described sensing chip; Described sensing chip comprises an optical clear substrate and the sensing membrane being located thereon, and described optical clear substrate contacts with the fluid media (medium) in described fluid prism, and described sensing membrane forms a bottom surface of described sample flow cell;
Described fluid prism is comprised of cavity and the fluid media (medium) that is positioned at chamber; The cavity of wherein said fluid prism is provided with two apertures and a macropore, and two apertures are respectively as input port and the delivery outlet of described fluid media (medium), and macropore is as the installing port of described sensing chip.
2. surface plasma body resonant vibration system according to claim 1, is characterized in that: the cavity of described fluid prism is made by optically transparent material;
The fluid media (medium) of described fluid prism is optically transparent liquid, gas or supercritical fluid.
3. surface plasma body resonant vibration system according to claim 1 and 2, is characterized in that: the material that surface plasma body resonant vibration can occur of serving as reasons of the sensing membrane in described sensing chip is made.
4. surface plasma body resonant vibration system according to claim 1 and 2, it is characterized in that: the light shaping that described incident beam shaping unit sends broadband polychromatic source is the adjustable p polarization parallel light of bulk or the little angle of p polarization converging beam, through described fluid prism, is incident in sensing chip.
5. surface plasma body resonant vibration system according to claim 1 and 2, is characterized in that: described incident beam shaping unit comprises optical alignment element, linear polarizer, adjustable diaphragm and lens.
6. surface plasma body resonant vibration system according to claim 1 and 2, is characterized in that: described reflected light detecting unit is by signal collection element and spectrum resolution installation composition; Described spectrum resolution device is the spectrograph that is equipped with the monochromator of photo-electric conversion element or is equipped with linear array photo-electric conversion element; Described signal collection element and spectrum resolution device direct-coupling or pass through coupling fiber.
7. utilize the method that in claim 1-6, the surface plasma body resonant vibration system described in any one detects, comprise the steps:
1) according to the refractive index selection of testing sample, can realize the refractive index of the fluid prism of surface plasma body resonant vibration;
2) testing sample is joined in the sample flow cell of the surface plasma body resonant vibration system described in any one in claim 1-6;
3) utilize the surface plasma body resonant vibration system described in any one in claim 1-6 to detect, obtain wavelength resonances surface plasma body resonant vibration curve.
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