CN101915750A - 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
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- CN101915750A CN101915750A CN2010102273749A CN201010227374A CN101915750A CN 101915750 A CN101915750 A CN 101915750A CN 2010102273749 A CN2010102273749 A CN 2010102273749A CN 201010227374 A CN201010227374 A CN 201010227374A CN 101915750 A CN101915750 A CN 101915750A
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- 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
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- 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 mechanism with development takes place, and novel drugs is found all to have great importance.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 China Patent No. ZL99107780.6, date of application 1999.5.28), the characteristics of this method are the precision height, but can only detect 1 sample at every turn, or detect several samples by the method for point by point scanning, can't realize that high flux measures.At 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 China Patent No. ZL200410057323.0), and basic comprising 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 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, enters Wollaston prism 107; Light is by 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 by 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 by measuring interference fringe, and in order to measure phase change delicately, each sensing point should comprise 3 interference fringes at least, needs to 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.
At 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 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 thereon 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 thereon 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 outs of phase of affix correspondence.Light after the modulation is by 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 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 by 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 described switching device shifter is alternatingly oriented in light path, thereby makes the P polarized component of collimated light and the phase differential between the S polarized component be 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 described reflected light by analyzer, thereby wherein P polarized component and S polarized component are interfered, produce interference image;
5) described interference image is imaged on the CCD target surface through imaging lens, and gathers the imaging interference image on the CCD target surface;
6), alternately gather the imaging interference image that produces behind additional 90 ° and-90 ° of phase differential by computer control wave plate switching mechanism;
7) gathering 2 two field pictures is 1 cycle, utilizes the 2 adjacent frame imaging interference images of additional 90 ° and-90 ° of phase differential that are collected, calculates the distribution function of reflection index distribution by following formula,
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 by 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, by 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.By 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 described 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 described incident arm.
According to one embodiment of present invention, wherein, be disposed with in the described polarization laser collimator apparatus: semiconductor laser, single-mode fiber, laser aligner and polarizing prism.
According to one embodiment of present invention, wherein, described wave plate switching device shifter comprises electric rotating machine, swing span, wave plate swivel mount and two quarter-wave plates, described quarter-wave plate is fixed in the wave plate swivel mount, described wave plate swivel mount is installed on the described swing span, described swing span is under the driving of described electric rotating machine, two described quarter-wave plates are positioned at the light path center in turn, make described quarter-wave plate vertical, to be implemented between P polarized component and S polarized component the phase differential of additional in turn 90 ° and-90 ° with optical axis.
According to one embodiment of present invention, wherein, the expansion bundle multiple of described parallel beam expand device is 2-8 times.
According to one embodiment of present invention, wherein, the principle of described bio-sensing unit by using surface plasma body resonant vibration, comprise: right angle or trapezoidal prism, sensing chip and place described sensing chip and described prism between index-matching fluid, the refractive index of wherein said prism, index-matching fluid and sensing chip substrate is identical, and described 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 by chemical method on described golden film surface.
According to one embodiment of present invention, wherein, described 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, described signal acquisition process unit comprises CCD or CMOS and computing machine, the described imaging interference image that collects from described CCD is transfused to described computing machine, and described computing machine utilizes the described imaging interference image of adjacent 2 frames to calculate index distribution.
Detection method by 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 by formula (1) and actual index of refraction relationship can be passed through fresnel formula emulation, and simulation result 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 is by the size decision of incident angle, by changing the measurement that incident angle can be implemented in the different refractivity interval.
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, described 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 described 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 by Green lens, and transmits the output of a segment distance in single-mode fiber 402.Then, by 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 by after the wave plate travel mechanism 405, and phase modulation (PM) has promptly been carried out in just alternately 90 ° of affixs and-90 ° phase differential between P polarization direction and S polarization direction.By 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, described wave plate travel mechanism 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 as shown in Figure 5.Quarter-wave plate 405-6 and pad 405-5 are fixed among the wave plate swivel mount 405-4 by 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, wave plate is vertical with optical axis, is implemented between P polarized component and S polarized component the phase differential of additional in turn 90 ° and-90 °.
According to one embodiment of present invention, described 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 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 the detected protein molecule of coupling.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 detected biomolecule when wherein flowing through, the interaction of molecules of biomolecule in the solution and chip surface coupling causes the chip surface structural change, thereupon Dui Ying 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, forms 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 described reflection arm successively.By the light of bio-sensing unit reflection, during by 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, described signal acquisition process unit comprises CCD 415 and is used for the computing machine of image acquisition and data analysis.Certainly, replaced C CD also can use CMOS.In described 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 the interference image that has added 90 ° and-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 of CCD 415 sensing correspondences 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 of CCD 415 sensing correspondences 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, determine the variations in refractive index of this point, draw corresponding measurement curve.So far, finished 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 by 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, by 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.By continuous coverage, just can obtain index distribution change information in time in real time.
Detection method by 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 the ordinary skill in the art, be appreciated that without departing from the principles and spirit of the present invention and can carry out multiple variation, modification, replacement and modification that scope of the present invention is by claims and be equal to and limit to these embodiment.
Claims (10)
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 by 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 described switching device shifter is alternatingly oriented in light path, thereby makes the P polarized component of collimated light and the phase differential between the S polarized component be 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 described reflected light by analyzer, thereby wherein P polarized component and S polarized component are interfered, produce interference image;
5) described interference image is imaged on the CCD target surface through imaging lens, and gathers the imaging interference image on the CCD target surface;
6), alternately gather the imaging interference image that produces behind additional 90 ° and-90 ° of phase differential by computer control wave plate switching mechanism;
7) gathering 2 two field pictures is 1 cycle, utilizes the 2 adjacent frame imaging interference images of additional 90 ° and-90 ° of phase differential that are collected, calculates the distribution function of reflection index distribution by following formula,
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.
2. the detection method of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 1 is characterized in that, and is further comprising the steps of:
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.
3. 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 described 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,
It is characterized in that, be disposed with polarization laser collimator apparatus, wave plate switching device shifter and parallel beam expand device in the described incident arm.
4. 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 described polarization laser collimator apparatus: semiconductor laser, single-mode fiber, laser aligner and polarizing prism.
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, described wave plate switching device shifter comprises electric rotating machine, swing span, wave plate swivel mount and two quarter-wave plates, described quarter-wave plate is fixed in the wave plate swivel mount, described wave plate swivel mount is installed on the described swing span, described swing span is under the driving of described electric rotating machine, two described quarter-wave plates are positioned at the light path center in turn, make described quarter-wave plate vertical, to be implemented between P polarized component and S polarized component the phase differential of additional in turn 90 ° and-90 ° with optical axis.
6. 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 described parallel beam expand device be 2-8 doubly.
7. 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 described bio-sensing unit by using surface plasma body resonant vibration, comprise: right angle or trapezoidal prism, sensing chip and place described sensing chip and described prism between index-matching fluid, the refractive index of wherein said prism, index-matching fluid and sensing chip substrate is identical, and described sensing chip surface is coated with the thick golden film of 30-50nm.
8. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 3 is characterized in that, wherein, is modified with the hydrophilic self-assembled film of one deck on described golden film surface by chemical method.
9. the detection system of the bio-molecular interaction based on the SPR interference imaging as claimed in claim 3 is characterized in that wherein, described 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.
10. 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, described signal acquisition process unit comprises CCD or CMOS and computing machine, the described imaging interference image that collects from described CCD is transfused to described computing machine, and described computing machine utilizes the described imaging interference image of adjacent 2 frames to calculate index distribution.
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| CN102253003A (en) * | 2011-06-08 | 2011-11-23 | 深圳大学 | Surface plasmon resonance sensing detection system and detection method thereof |
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| 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 |
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