CN101915750B - Biomolecule interaction detection method and system based on SPR interference imaging - Google Patents
Biomolecule interaction detection method and system based on SPR interference imaging Download PDFInfo
- Publication number
- CN101915750B CN101915750B CN2010102273749A CN201010227374A CN101915750B CN 101915750 B CN101915750 B CN 101915750B CN 2010102273749 A CN2010102273749 A CN 2010102273749A CN 201010227374 A CN201010227374 A CN 201010227374A CN 101915750 B CN101915750 B CN 101915750B
- Authority
- CN
- China
- Prior art keywords
- imaging
- interference
- bio
- interference image
- wave plate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
- G01N21/45—Refractivity; Phase-affecting properties, e.g. optical path length using interferometric methods; using Schlieren methods
Abstract
The invention discloses a biomolecule interaction detection method comprising the following steps: 1) obtaining collimated light; 2) alternately modulating the phase difference between the P polarized component and the S polarized component of the collimated light to 90 degrees or -90 degrees; 3) allowing phase modulation light to enter into a biological sensing unit so as to stimulate surface plasma resonance, and reflecting light; 4) interfering the P polarized component with the S polarized component in the reflected light to generate an interference image; 5) imaging the interference image, and collecting the imaging interference image; 6) alternately collecting the imaging interference image generated after adding phase differences of 90 degrees and -90 degrees; and 7) utilizing the collected adjacent two frames of imaging interference images to calculate a distribution function reflecting the distribution of refractive index. The invention can continuously measure to obtain the refractive index distribution change information along with time in real time.
Description
Technical field
The present invention relates to biological technical field, particularly be used for realizing the detection method and the detection system thereof of bio-molecular interactions such as high flux, high precision, unmarked, real-time sensor matter-protein, protein-DNA, protein-effector molecules, Ag-Ab, ligand-receptor, medicine-target.
Background technology
The essence of life process is the process of various bio-molecular interactions, and the generation of disease and change procedure also concern with interaction of molecules invariably.Just because of this, the detection of bio-molecular interaction has become one of research focus of life science.Wherein, protein is the expression of gene product, is carrying out the function of life, between the research protein molecule and with other interaction of molecules, to illustrating life process and disease the mechanism with development takes place, and the novel drugs discovery has great importance all.Compare with gene, protein not only kind is more, and space-time characterisation is arranged, and the method that detects gene is difficult to satisfy the requirement of protein detection, thereby needs high flux, high precision, unmarked, the real-time novel detection method that also can keep protein active in a hurry.
Based on surface plasma body resonant vibration (surface plasmon resonance; Be called for short SPR) biology sensor be a kind of optical detecting method with high sensitivity, characteristics such as real-time, unmarked, be considered to the more satisfactory method of detection of biological interaction of molecules.The detection method of surface plasmon resonance biosensor mainly contains angle scanning, light intensity detection and phase-detection etc., and wherein phase-detection is sensitive higher.The inventor herein had once proposed based on the difference interference biomolecular interaction real-time phase detection analysis method of zeeman laser and system (referring to Chinese patent ZL99107780.6; Date of application 1999.5.28); The characteristics of this method are that precision is high; But can only detect 1 sample at every turn, or detect several samples, can't realize the high flux measurement through the method for point by point scanning.To this deficiency, the inventor of this patent has proposed again based on the method and system of space phase interferometric modulator array detection of biological chip (referring to Chinese patent ZL200410057323.0), and basic comprising is as shown in Figure 1.The linear polarization directional light that helium-neon laser 101 sends sees through prism 102, index-matching fluid 103 and sensing chip substrate of glass 104, incides substrate of glass 104 and is plated in the interface between the suprabasil golden film 105, and the thickness of golden film is 30-50nm.When incident angle was in resonance angle, incident light was at golden film surface excitation surface plasma body resonant vibration.Light reflects from substrate of glass 104 and the interface that is plated between the suprabasil golden film 105, sees through one dimension beam expanding lens 106, gets into Wollaston prism 107; Light is through Wollaston prism 107 time, and P polarized component wherein and S polarized component are separated a very little angle, then penetrates from Wollaston prism 107.Then, light is when the polarizing prism 108, and P polarized component S polarized component interferes, and interference fringe is imaged on through imaging len 109 on the target surface of CCD 110, by the computer acquisition interference image and handle.If the refractive index on golden film surface changes, P polarized light in the reflected light and the phase differential between the S polarized light change thereupon, are imaged on promptly corresponding the moving of interference fringe on the CCD.Utilize Fourier's fringe analysis technology that the interference image of being gathered is handled, resolve phase change, obtain the variations in refractive index information of chip surface, thereby realized array detection.This method is to obtain phase change through measuring interference fringe, and in order to measure phase change delicately, each sensing point should comprise 3 interference fringes at least, need occupy 12 adjacent CCD pixels so at least.Obviously, the row of CCD and the pixel of row are all limited, detect flux and are restricted naturally.
To above-mentioned deficiency, the inventor herein has further proposed the surface plasma volume array bio-sensing method (referring to Chinese patent ZL200510086332.7) based on the interference of time domain phase modulation (PM), and ultimate principle is as shown in Figure 2.The directional light that light source 201 sends sees through prism 202 and refractive index match layer 203, incide on the substrate of glass 204 and the interface between the plating golden film 205 above that of sensing chip, and from this boundary reflection, the thickness of golden film is 30-50nm.When incident light is on the resonance angle, excite the plasma resonance on golden film surface, the phase place of the light of launching from the interface between substrate of glass 204 and the golden film 205 is acute variation with the change of golden film surface dielectric refractive index.Reflected light is injected electro-optic crystal 207 after seeing through refractive index match layer 203, prism 202 and one dimension beam expanding lens 206.Electro-optic crystal 207 can apply 5 kinds of different voltages with different above that by 208 controls of high-voltage modulation power supply.Thereupon, after light passes through electro-optic crystal 207, just poor between catoptrical P and the S polarized component by 5 corresponding outs of phase of affix.Light after the modulation is through polarizing prism 209 time, and P polarized light and S polarized light interfere, and interference image is imaged on the target surface of CCD 211 by imaging len 210.The corresponding voltage of the collection of CCD 211 images and high-voltage power supply 208 outputs is synchronous, and 5 different voltages with different of high-voltage power supply 208 outputs are 1 cycle, and corresponding collection 5 two field pictures also are 1 cycle.High-voltage power supply 208 output voltage that can circulate, corresponding interference image also can circle collection.Then, utilize the Hariharan algorithm, 5 frame interference images under 5 adjacent voltages of being gathered are resolved, obtain instant catoptrical phase change.In theory, utilize this method, only need the pixel of a CCD, just can detect a sensing point on the sensing chip, can realize high throughput testing.Yet, when on electro-optic crystal, applying high voltage, inverse piezoelectric effect can appear thereupon; Cause the deformation of the structure generation microscale of electro-optic crystal; Cause the light path minor shifts, promptly facula position moves, and directly produces measuring error; The consistance that influences array detection is poor, is difficult to reach the requirement of high Precision Detection.This is the intrinsic defective of electro-optic crystal, can't remedy.
Summary of the invention
The objective of the invention is to overcome the deficiency of above-mentioned technology; A kind of new surface plasma resonance sensing systems approach and system based on interference imaging are provided; Can high flux, high precision, real-time, unmarked detection protein-chip, obtain interactional information between the protein.
A kind of detection method of the bio-molecular interaction based on the SPR interference imaging is provided according to an aspect of the present invention, has it is characterized in that, may further comprise the steps:
1) after the light that semiconductor laser is sent passes through optical fiber coupling and collimation,, obtains the collimated light beam of the adjustable linear polarization in polarization direction again through polarizing prism;
2) described linear polarization collimated light beam is passed through by computer-controlled quarter-wave plate switching device shifter; The quarter-wave plate of 2 quick shaft direction quadratures in the said switching device shifter is alternatingly oriented in light path, thereby the P polarized component and the phase differential between the S polarized component that make collimated light are become 90 ° or-90 ° by modulation alternately;
3) collimated light through phase modulation (PM) is reentered on the substrate of glass and the interface between the golden film that is mapped to the sensing chip in the bio-sensing unit, thereby excitating surface plasma resonates, with the time from this boundary reflection;
4) make said reflected light through analyzer, thereby wherein P polarized component and S polarized component are interfered, produce interference image;
5) said interference image is imaged on the CCD target surface through imaging lens, and gathers the imaging interference image on the CCD target surface;
6) by computer control wave plate switching mechanism, alternately gather additional 90 ° with-90 ° of phase differential after the imaging interference image that produces;
7) gathering 2 two field pictures is 1 cycle, utilizes the additional 90 ° of adjacent 2 frames imaging interference images with-90 ° of phase differential that collected, through the distribution function of computes reflection index distribution,
In the formula, I
+Be the refractive index of the interference image of additional 90 ° of phase differential, I
-Be the refractive index of the interference image of additional-90 ° of phase differential, A is the distribution function of reflection index distribution.
According to one embodiment of present invention, further comprising the steps of:
8) repeating step 2) to the operation of step 7), circle collection is calculated to a series of imaging interference images, and draw, thereby obtain the signal graph of a series of lasting reflection index distribution with a series of distribution functions and the refractive index of calculating gained.
Detection method through the bio-molecular interaction based on the SPR interference imaging of the present invention; Based on surface plasma body resonant vibration array bio-sensing principle; Between the P of incident light polarized component and S polarized component, additional in turn 90 ° and-90 ° of phase differential obtain corresponding interference image; Through 2 width of cloth are resolved interference image, can access 1 reaction index distribution frame.Utilize 2 adjacent frame interference images, can calculate the a-signal figure of 1 reflection index distribution; If gather n frame interference image altogether, then can calculate the signal graph A that n-1 opens the reflection index distribution.Through continuous coverage, just can obtain index distribution change information in time in real time.
A kind of detection system of the bio-molecular interaction based on the SPR interference imaging also is provided according to a further aspect in the invention, has comprised: produced collimated light and it is replaced the incident arm of additional phase error; Acceptance is through the collimated light of additional phase error and the bio-sensing unit of excitating surface plasma resonance; Acceptance is by the light of said bio-sensing unit reflection, it produced interfere and with the reflection arm of interference image imaging; And obtain interference image and carry out the signal acquisition process unit of subsequent treatment, wherein, be disposed with polarization laser collimator apparatus, wave plate switching device shifter and parallel beam expand device in the said incident arm.
According to one embodiment of present invention, wherein, be disposed with in the said polarization laser collimator apparatus: semiconductor laser, single-mode fiber, laser aligner and polarizing prism.
According to one embodiment of present invention; Wherein, Said wave plate switching device shifter comprises electric rotating machine, swing span, wave plate swivel mount and two quarter-wave plates, and said quarter-wave plate is fixed in the wave plate swivel mount, and said wave plate swivel mount is installed on the said swing span; Said swing span is under the driving of said electric rotating machine; Two said quarter-wave plates are positioned at the light path center in turn, make said quarter-wave plate vertical with optical axis, be implemented between P polarized component and S polarized component additional in turn 90 ° with-90 ° phase differential.
According to one embodiment of present invention, wherein, the expansion bundle multiple of said parallel beam expand device is 2-8 times.
According to one embodiment of present invention; Wherein, The principle of said bio-sensing unit by using surface plasma body resonant vibration; Comprise: right angle or trapezoidal prism, sensing chip and place said sensing chip and said prism between index-matching fluid, the refractive index of wherein said prism, index-matching fluid and sensing chip substrate is identical, and said sensing chip surface is coated with the thick golden film of 30-50nm.
According to one embodiment of present invention, wherein, be modified with the hydrophilic self-assembled film of one deck through chemical method on said golden film surface.
According to one embodiment of present invention, wherein, said reflection arm comprises polarizing prism and imaging lens, and this camera lens is imaged on the interference image on golden film surface on the CCD target surface.
According to one embodiment of present invention; Wherein, Said signal acquisition process unit comprises CCD or CMOS and computing machine; The said imaging interference image that collects from said CCD is transfused to said computing machine, and said computing machine utilizes the said imaging interference image of adjacent 2 frames to calculate index distribution.
Detection method through this bio-molecular interaction based on the SPR interference imaging provided by the invention and the detection system that realizes this method based on the bio-molecular interaction of SPR interference imaging; Can high flux, high precision, real-time, unmarked detection; Obtain the information of bio-molecular interaction; Offer biomedicine and pharmaceuticals researcher, carry out protein science research, medical diagnosis on disease, drug discovery and exploitation etc.
Description of drawings
Fig. 1 is a kind of SPR space phase modulation detection principle schematic of prior art;
Fig. 2 is that a kind of SPR time domain phase modulation of prior art detects principle schematic;
Fig. 3 is the theoretical foundation of the computing method of one embodiment of the present of invention;
Fig. 4 is the synoptic diagram based on the detection system structure of the bio-molecular interaction of SPR interference imaging of one embodiment of the present of invention;
Fig. 5 is the wave plate switching device shifter structural representation of one embodiment of the present of invention, and wherein Fig. 5 a is a front view, and Fig. 5 b is a side view;
Fig. 6 is the detection of one embodiment of the present of invention and the process flow diagram of signal processing.
Embodiment
Below in conjunction with accompanying drawing, to the detection method of the bio-molecular interaction based on the SPR interference imaging of the present invention and be used to realize that the detection system based on the bio-molecular interaction of SPR interference imaging of this method describes for example.
Computing method of the present invention are to utilize the catoptrical amplitude of SPR and the phase place characteristic with the refractive index acute variation; The signal graph A of the reflection index distribution that calculates through formula (1) can pass through fresnel formula emulation with actual index of refraction relationship, and simulation result is as shown in Figure 3.As can be seen from the figure, both are linear in certain interval.Therefore, the variation of A can reflect actual variations in refractive index, and the position between linear zone can be implemented in the interval measurement of different refractivity by the size decision of incident angle through changing incident angle.
Fig. 4 has shown the structural representation based on the detection system of the bio-molecular interaction of SPR interference imaging of one embodiment of the present of invention.In the detection system of present embodiment, comprise four major parts such as incident arm, bio-sensing unit, reflection arm and signal acquisition process unit.
According to one embodiment of present invention; Said incident arm is positioned at a side of bio-sensing unit, comprises temperature control semiconductor laser 401, single-mode fiber 402, laser aligner 403, polarizing prism 404, wave plate rotating disk 405, electric rotating machine 406 and laser beam expanding lens 407 successively.
According to one embodiment of present invention; In said incident arm, temperature control semiconductor laser 401 sends the laser of stabilized intensity under the driving of constant-current supply; Light is coupled in the single-mode fiber 402 through Green lens, and in single-mode fiber 402, transmits the output of a segment distance.Then, through becoming the collimated light beam that hot spot is 3-10mm after the collimating apparatus 403, see through polarizing prism 404 again, become linearly polarized light, the polarization direction is adjustable.Linearly polarized light is through after the wave plate travel mechanism 405, and just the phase differential of 90 ° of affixs and-90 ° alternately between P polarization direction and S polarization direction has promptly carried out phase modulation (PM).Through laser beam expanding lens 407, expanding the Shu Chengwei diameter is the collimated light beam of 10-30mm, is mapped in the bio-sensing unit again through the light beam after the phase modulation (PM).
According to one embodiment of present invention, said wave plate travel mechanism is as shown in Figure 5, comprises that electric rotating machine 405-1, swing span 405-2, hold-down ring 405-3, wave plate swivel mount 405-4, pad 405-5 and quarter-wave plate 405-6 form.Quarter-wave plate 405-6 and pad 405-5 are fixed among the wave plate swivel mount 405-4 through hold-down ring 405-3; Wave plate swivel mount 405-4 is installed on the swing span 405-2; Swing span 405-2 is under the driving of electric rotating machine 405-1; 2 wave plate 405-6 are positioned at the light path center in turn, and wave plate is vertical with optical axis, be implemented between P polarized component and S polarized component additional in turn 90 ° with-90 ° phase differential.
According to one embodiment of present invention; Said bio-sensing unit is a surface plasma resonance biosensor; Comprise right-angle prism 408, sensing chip 410 and place sensing chip and prism between index-matching fluid 409, chip surface be coated with 40nm gold film 411 and be positioned at the fluid pool 412 below the sensing chip 410, sensing chip 410 is close to fluid pool 412 and is sealed.
In bio-sensing of the present invention unit, prism 408 uses identical K9 or ZF5 glass with the substrate of glass of sensing chip 410, and the refractive index of index-matching fluid 409 is consistent with the glass matrix refractive index of prism 408 and chip 410.Chip 410 is coated with the golden film 411 of 30-50nm on substrate of glass, golden film 411 surfaces use chemical method to modify the hydrophilic self-assembled film of one deck, are used for coupling protein molecule to be detected.Fluid pool 412 is close to golden film 411 and is sealed, and the surface of golden film 411 becomes the wherein wall of one side of fluid pool.When the solution of biomolecule to be detected when wherein flowing through, the interaction of molecules of biomolecule in the solution and chip surface coupling causes the chip surface structural change, corresponding thereupon variations in refractive index.Collimated light beam sees through prism 408, index-matching fluid 409 and chip 410, and in the total reflection of generation at the interface of chip 410 and golden film 411, when incident angle was resonance angle, the energy of incident light was coupled to golden film 411 by evanescent wave, formed surface plasma body resonant vibration.
According to one embodiment of present invention, the opposite side that is positioned at the bio-sensing unit is provided with the reflection arm, comprises polarizing prism 413 and imaging lens 414 in the said reflection arm successively.By the light of bio-sensing unit reflection, during through the polarizing prism 413 on the reflection arm, P polarized light and S polarized light interfere, and interference image is imaged on the CCD 415 by imaging lens 414, and this interference image is the real image of sensing surface.
According to one embodiment of present invention, said signal acquisition process unit comprises CCD 415 and the computing machine that is used for IMAQ and data analysis.Certainly, replaced C CD also can use CMOS.In said signal acquisition process unit, the interference image that collects from CCD 415 is transfused to computing machine, handles in real time.The collection of the motion of electric rotating machine 405-1 and CCD 415 is all by computer control, alternately obtains and added 90 ° of interference images with-90 ° of phase differential.Computing machine utilizes 2 adjacent frame interference images, can calculate the a-signal figure of 1 reflection index distribution, if gather n frame interference image altogether, then can calculate the a-signal figure that n-1 opens the reflection index distribution.Clearly, compare, resolve efficient and improved 4 times with five traditional step phase-shift methods.Acquisition time between the 2 adjacent two field pictures is little at interval; Can think that incident intensity is constant when gathering 2 adjacent frame interference images; The signal distribution plots of the reflection variations in refractive index that calculates like this and initial light distribution are irrelevant; The error of having avoided the unevenness of light distribution to bring can also suppress the measuring error that the light intensity drift causes simultaneously.
Specify detection according to an embodiment of the invention and signal processing below in conjunction with accompanying drawing.Fig. 6 is the detection of one embodiment of the present of invention and the process flow diagram of signal processing.
When measuring beginning, computing machine is at first controlled electric rotating machine 405-1, and a wave plate is positioned at the light path center, 90 ° or-90 ° phase differential of affix between P and S polarized component, and the interference image that CCD 415 sensings are corresponding is read in by computing machine.
And then, electric rotating machine 405-1 is positioned at the light path center with another piece wave plate, affix-90 ° or 90 ° of phase differential between P and S component, and the interference image that CCD 415 sensings are corresponding is read in by computing machine.Thereafter, computing machine utilizes 2 frame interference images, according to formula (1), calculates the A distribution function of reflection index distribution, and to obtain 1 thus be the signal graph of ordinate and horizontal ordinate with distribution function A and refractive index respectively.Simultaneously, can also select the sensing point on the chip, confirm the variations in refractive index of this point, draw corresponding measurement curve.So far, accomplished an operating cycle.
Repeat the aforesaid operations process, can obtain the signal graph of 1 new reflection index distribution again.Calculate the difference between these 2 signal graphs, the variations in refractive index that causes when just obtaining on corresponding sensing point, interaction of molecules taking place.
Circulation is carried out, the just interaction of detection of biological molecule in real time, and draw the time dependent curve of refractive index of corresponding sensing point, offer the biomedical research personnel then and carry out related resolution.
Detection method through the bio-molecular interaction based on the SPR interference imaging of the present invention; Based on surface plasma body resonant vibration array bio-sensing principle; Between the P of incident light polarized component and S polarized component, additional in turn 90 ° and-90 ° of phase differential obtain corresponding interference image; Through 2 width of cloth interference images are calculated, can access 1 is the signal graph of ordinate and horizontal ordinate with distribution function A and refractive index respectively.Utilize 2 adjacent frame interference images, can calculate the signal graph of 1 reflection index distribution; If gather n frame interference image altogether, then can calculate the signal graph that n-1 opens the reflection index distribution.Through continuous coverage, just can obtain index distribution change information in time in real time.
Detection method through this bio-molecular interaction based on the SPR interference imaging provided by the invention and the detection system that realizes this method based on the bio-molecular interaction of SPR interference imaging; Can high flux, high precision, real-time, unmarked detection; Obtain the information of bio-molecular interaction; Offer biomedicine and pharmaceuticals researcher, carry out protein science research, medical diagnosis on disease, drug discovery and exploitation etc.
Although illustrated and described embodiments of the invention; For those of ordinary skill in the art; Be appreciated that under the situation that does not break away from principle of the present invention and spirit and can carry out multiple variation, modification, replacement and modification that scope of the present invention is accompanying claims and be equal to and limit to these embodiment.
Claims (7)
1. the detection method based on the bio-molecular interaction of SPR interference imaging is characterized in that, may further comprise the steps:
1) after the light that semiconductor laser is sent passes through optical fiber coupling and collimation,, obtains the collimated light beam of the adjustable linear polarization in polarization direction again through polarizing prism;
2) said linear polarization collimated light beam is passed through by computer-controlled quarter-wave plate switching device shifter; The quarter-wave plate of 2 quick shaft direction quadratures in the said quarter-wave plate switching device shifter is alternatingly oriented in light path, thereby the P polarized component and the phase differential between the S polarized component that make collimated light are become 90 ° or-90 ° by modulation alternately;
3) collimated light through phase modulation (PM) is reentered on the substrate of glass and the interface between the golden film that is mapped to the sensing chip in the bio-sensing unit, thereby excitating surface plasma resonance make light from this boundary reflection simultaneously;
4) make said reflected light through analyzer, thereby wherein P polarized component and S polarized component are interfered, produce interference image;
5) said interference image is imaged on the CCD target surface through imaging lens, and gathers the imaging interference image on the CCD target surface;
6) by computer control quarter-wave plate switching device shifter, alternately gather additional 90 ° with-90 ° of phase differential after the imaging interference image that produces;
7) gathering 2 two field pictures is 1 cycle, utilizes the additional 90 ° of adjacent 2 frames imaging interference images with-90 ° of phase differential that collected, through the distribution function of computes reflection index distribution,
In the formula, I
+Be the refractive index of the interference image of additional 90 ° of phase differential, I
-Be the refractive index of the interference image of additional-90 ° of phase differential, A is the distribution function of reflection index distribution;
8) repeating step 2) to the operation of step 7), to circle collection to a series of imaging interference images calculate, and draw, thereby obtain the signal graph of a series of lasting reflection index distribution with a series of distribution functions and the refractive index of calculating gained.
2. the detection system based on the bio-molecular interaction of SPR interference imaging comprises: produce collimated light
And it is replaced the incident arm of additional phase error;
Acceptance is through the collimated light of additional phase error and the bio-sensing unit of excitating surface plasma resonance;
Acceptance is by the light of said bio-sensing unit reflection, it produced interfere and with the reflection arm of interference image imaging; And
Obtain interference image and carry out the signal acquisition process unit of subsequent treatment; Said signal acquisition process unit comprises CCD or CMOS and computing machine; The said imaging interference image that collects from said CCD is transfused to said computing machine; Said computing machine utilizes the said imaging interference image of adjacent 2 frames to calculate index distribution
It is characterized in that; Be disposed with polarization laser collimator apparatus, quarter-wave plate switching device shifter and parallel beam expand device in the said incident arm; Said quarter-wave plate switching device shifter comprises electric rotating machine, swing span, wave plate swivel mount and two quarter-wave plates, and said quarter-wave plate is fixed in the wave plate swivel mount, and said wave plate swivel mount is installed on the said swing span; Said swing span is under the driving of said electric rotating machine; Two said quarter-wave plates are positioned at the light path center in turn, make said quarter-wave plate vertical with optical axis, be implemented between P polarized component and S polarized component additional in turn 90 ° with-90 ° phase differential.
3. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 3 is characterized in that, is disposed with in the said polarization laser collimator apparatus: semiconductor laser, single-mode fiber, laser aligner and polarizing prism.
4. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 3 is characterized in that, the expansion bundle multiple of said parallel beam expand device be 2-8 doubly.
5. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 3; It is characterized in that; Wherein, the principle of said bio-sensing unit by using surface plasma body resonant vibration comprises: right angle or trapezoidal prism, sensing chip and place said sensing chip and said prism between index-matching fluid; The refractive index of wherein said prism, index-matching fluid and sensing chip substrate is identical, and said sensing chip surface is coated with the thick golden film of 30-50nm.
6. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 5 is characterized in that, wherein, is modified with the hydrophilic self-assembled film of one deck on said golden film surface through chemical method.
7. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 5 is characterized in that wherein, said reflection arm comprises polarizing prism and imaging lens, and this camera lens is imaged on the interference image on golden film surface on the CCD target surface.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102273749A CN101915750B (en) | 2010-07-07 | 2010-07-07 | Biomolecule interaction detection method and system based on SPR interference imaging |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102273749A CN101915750B (en) | 2010-07-07 | 2010-07-07 | Biomolecule interaction detection method and system based on SPR interference imaging |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101915750A CN101915750A (en) | 2010-12-15 |
| CN101915750B true CN101915750B (en) | 2012-01-18 |
Family
ID=43323311
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010102273749A Expired - Fee Related CN101915750B (en) | 2010-07-07 | 2010-07-07 | Biomolecule interaction detection method and system based on SPR interference imaging |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101915750B (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102243174B (en) * | 2011-04-14 | 2013-05-22 | 中国科学院半导体研究所 | Phase detection-based surface plasma resonance sensing device |
| CN102353654B (en) * | 2011-06-08 | 2014-06-25 | 深圳大学 | Surface plasma resonance sensing detection system and its detection method |
| CN102253003B (en) * | 2011-06-08 | 2014-05-28 | 深圳大学 | Surface plasmon resonance sensing detection system and detection method thereof |
| CN102621686B (en) * | 2012-03-19 | 2014-02-19 | 浙江大学 | Prism phase modulator for SPR (Surface Plasmon Resonance) sensing and modulation method thereof |
| CN103743674A (en) * | 2013-09-30 | 2014-04-23 | 北京航空航天大学 | Method and system for enhancing non-specular reflection effect |
| CN105806800B (en) * | 2014-12-30 | 2019-01-22 | 深圳先进技术研究院 | Terahertz light fiber sensing equipment and the contamination detection method for utilizing the device |
| CN104596989B (en) * | 2015-01-23 | 2017-05-31 | 四川大学 | Method based on interference fringe picture treatment measurement refractive index of transparent medium distribution |
| CN108572160B (en) * | 2017-03-14 | 2020-05-19 | 华中科技大学 | Refractometer for measuring refractive index distribution |
| CN110927121B (en) * | 2019-12-05 | 2022-07-05 | 深圳大学 | Phase type SPR detection device and method based on white light interference spectrum |
| CN111487190B (en) * | 2020-04-20 | 2021-08-31 | 中国科学院力学研究所 | Single-ion imaging detection method and device |
| CN113030028A (en) * | 2021-04-20 | 2021-06-25 | 浙江科技学院 | Method for detecting two-dimensional space phase information by using SPR phase shift interference |
| CN114414531B (en) * | 2022-01-30 | 2023-07-28 | 福州大学 | In-situ on-line detection method and device for organoid metabolic molecules |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030219809A1 (en) * | 2002-03-26 | 2003-11-27 | U-Vision Biotech, Inc. | Surface plasmon resonance shifting interferometry imaging system for biomolecular interaction analysis |
| JP4150966B2 (en) * | 2003-08-27 | 2008-09-17 | 東洋紡績株式会社 | Surface plasmon resonance measurement method |
| CN1312476C (en) * | 2004-08-27 | 2007-04-25 | 清华大学 | Method and system for detecting biological chip by space phase modulation interference array |
| US7365855B2 (en) * | 2005-07-08 | 2008-04-29 | The Chinese University Of Hong Kong | Optical sensing devices with SPR sensors based on differential phase interrogation and measuring method using the same |
| CN100347546C (en) * | 2005-09-02 | 2007-11-07 | 清华大学 | Sensing method of protein chip and detection system therefor |
-
2010
- 2010-07-07 CN CN2010102273749A patent/CN101915750B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN101915750A (en) | 2010-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101915750B (en) | Biomolecule interaction detection method and system based on SPR interference imaging | |
| US10578554B2 (en) | Spectrum-scanned SPR imaging detection system | |
| US7084980B2 (en) | SPR interferometer | |
| US7339681B2 (en) | Surface plasmon resonance microscope using common-path phase-shift interferometry | |
| CN100347546C (en) | Sensing method of protein chip and detection system therefor | |
| KR101983400B1 (en) | Systems and methods for detection and imaging of two-dimensional sample arrays | |
| CN1312476C (en) | Method and system for detecting biological chip by space phase modulation interference array | |
| CN106896095B (en) | The micro-imaging technique of composite surface plasma resonance and surface-enhanced Raman | |
| CN104849481B (en) | A kind of full-automatic high flux optical bio sensing device | |
| CN104155266A (en) | Multichannel parallel detection surface plasma resonance biosensor, and preparation and detection methods thereof | |
| CN108709874B (en) | Surface plasma resonance sensing device and method based on self-mixing interference | |
| US20130164861A1 (en) | Biological molecule detecting apparatus and biological molecule detecting method | |
| CN102042972B (en) | Interference imaging method and system thereof for real-time detection of interaction of biological molecules | |
| CN109115728B (en) | Surface electromagnetic mode resonance hyperspectral imaging device, imaging method and application | |
| Chen et al. | Fast spectral surface plasmon resonance imaging sensor for real-time high-throughput detection of biomolecular interactions | |
| CN100451622C (en) | Method and system for testing heterodyne phase of resonance biochemical multichannel of surface plasma | |
| CN107356560B (en) | Total reflection type oblique incident light reflection difference scanning imaging device and using method thereof | |
| US20140227136A1 (en) | Surface plasmon resonance biosensor system | |
| Yu et al. | A novel surface plasmon resonance imaging interferometry for protein array detection | |
| Zhang et al. | Label-Free Imaging of Single Proteins and Binding Kinetics Using Total Internal Reflection-Based Evanescent Scattering Microscopy | |
| CN109883350A (en) | A kind of high precision measuring system and measurement method of abnormal curved surface inside configuration pattern | |
| Huang et al. | Recent advances in surface plasmon resonance microscopy | |
| Sutapun et al. | A multichannel surface plasmon resonance sensor using a new spectral readout system without moving optics | |
| CN207181294U (en) | Total-reflection type oblique incident ray difference in reflection scanned imagery device | |
| CN111208066B (en) | Biological detection device and method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120118 Termination date: 20120707 |