CN107219199A - Novel angle modulation SPR imaging systems based on 4F systems - Google Patents
Novel angle modulation SPR imaging systems based on 4F systems Download PDFInfo
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- 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
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- 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
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Abstract
SPR imaging systems are modulated the invention discloses a kind of novel angle based on 4F systems, when scanning galvanometer carries out angle scanning, it is incident to incidence and outgoing of the P polarization light by keeping parallelism of the first 4F systems, no matter how the angle of incident light changes, it is irradiated to the facula position formed in sensing membrane and remains constant, and the 2nd 4F systems are added in reflected light path, it ensure that under a different angle, the image space of receiver is not moved, stability is good and image quality is high, so as to ensure in scanning process that it is identical all the time and constant with search coverage that SPR is excited, the sensitive zones in sensing membrane can be increased, and cause sensitive face that there is good sensing uniformity, quick detection can effectively be realized, and meet the requirement of the highly sensitive detection of quick detection process.Further, said system also has the advantages that simple in construction, small volume.
Description
Technical field
The present invention relates to picture imaging techniques field, more particularly to a kind of novel angle based on 4F systems modulate SPR into
As system.
Background technology
Surface plasma resonance (Surface Plasmon Resonance, SPR) with its highly sensitive, high flux, exempt from mark
The many advantages such as note, quick detection are applied to many biochemical fields such as biological detection, drugs analysis, molecular recognition,
Quick development has been obtained in the late two decades, has become a kind of important monitoring instrument.
Kretschmann prism structures are the most frequently used coupled structures of current spr sensor, mainly have four based on the structure
Detection pattern is planted, is intensity modulation type, angle modulated type, wavelength modulation type and phase modulation-type respectively.Wherein, intensity modulated
SPR technique needs to be operated in fixed excitation wavelength and incident angle, and (high flux detection) is detected simultaneously for different kinds of molecules
Using the corresponding sensitivity of each sensing site is different, and spr signal response uniformity is looked into, and causes flase drop or missing inspection, and the technology pair
Sensing chip requires high, causes cost of use high.And angle modulated SPR has bigger dynamic range, it can reduce to sensing
The requirement of chip, is greatly reduced use cost.But angle modulated SPR needs mechanical scanning to enter arm and feeler arm, to instrument knot
Structure design requirement is very high, and needs bulky scanning mechanism, due to there is the mechanical part motion scan of big quality,
Cause that detection speed is slow, angular resolution is low, complicated and stability is poor, it is impossible to meet the highly sensitive detection of Fast Process
Demand.
The content of the invention
It is a primary object of the present invention to provide a kind of novel angle based on 4F systems to modulate SPR imaging systems, it is intended to
Solve the technical problem that detection speed is slow in the prior art, sensitivity is low and stability is poor.
To achieve the above object, first aspect present invention provides a kind of novel angle modulation SPR imagings based on 4F systems
System, including input path, sensing module and reflected light path；
The input path includes：Light source module, the polarizer, scanning galvanometer and the first 4F systems；
The sensing module includes prism, sensing membrane and flow cell, and the flow cell is used to sample to be tested flowing through institute
State sensing membrane；
The reflected light path includes the 2nd 4F systems and receiver；
The light source module sends collimated light by the polarizer, forms P polarization light, and the P polarization light is incident to institute
State scanning galvanometer to reflect, the P polarization light of reflection is incident to the first 4F systems, passes through after the first 4F system exits
The prism-coupled, and incide the sensing membrane of prism surface and excite SPR phenomenons, the sensing membrane is by reflected P polarized light, reflection
P polarization light be incident to the 2nd 4F systems, by the 2nd 4F system exits to the receiver, by the receiver shape
Into the SPR angular spectrum curves of all sensing sites, and terminal device is fed back to, the terminal device utilizes the SPR angular spectrums curve
Generate SPR images.
Further, the first 4F systems include the first lens and the second lens, the P polarization of the scanning galvanometer reflection
Light is projected from first lens entrance and from second lens.
Further, the 2nd 4F systems include the 3rd lens and the 4th lens, and the P polarization of sensing membrane reflection
Light is projected from the 3rd lens entrance and from the 4th lens.
Further, the system also includes analyzer, and the analyzer is saturating positioned at the 3rd lens and the described 4th
Between mirror, for the S-polarization light for suppressing to produce in scattering process.
Further, the scanning galvanometer is rotated according to preset step-length, to change the P for being incident to the first 4F systems
The incident direction of polarised light, the scope of the incident direction is [- θ ,+θ].
Further, the receiver is detector array.
Further, the detector array is electronic CCD or cmos detector or array photodetectors.
Further, the light source module is wavelengthtunable light source module.
Further, the light source module includes wideband light source and wave filter；Passed through after the wideband light source launching beam
The filter filtering is crossed, the collimated light and the incident polarizer is obtained；
The wideband light source is Coherent Wideband light source, non-coherent broad band light source or partially coherent wideband light source, the filter
Ripple device is liquid crystal filter, acousto-optic filter or the wave filter being made up of multigroup optical filter.
Further, the light source module is made up of the light source of at least two independent different wave lengths.
The present invention provide it is a kind of based on 4F systems novel angle modulation SPR imaging systems, the system include input path,
Sensing module and reflected light path, the input path include light source module, the polarizer, scanning galvanometer and the first 4F systems, the sensing
Module includes prism, sensing membrane and flow cell, and the sensing membrane is located between flow cell and prism, and the flow cell is used for will be to be tested
Sample flows through the sensing membrane, and the reflected light path includes the 2nd 4F systems and receiver.Within the system, light source module is sent
Collimated light passes through the polarizer, forms P polarization light, and P polarization light is incident to scanning galvanometer and launched, and the P polarization light of reflection is incident
To the first 4F systems, by prism-coupled after the first 4F system exits, and incide the sensing membrane of prism surface excite SPR show
As the sensing membrane will launch P polarization light, and the P polarization light of reflection is incident to the 2nd 4F systems, by the 2nd 4F system exits to reception
Device, is formed the SPR angular spectrum curves of all sensing sites by the receiver, and feeds back to terminal device, and the terminal device utilizes institute
There are the SPR angular spectrums curve generation SPR images of sensing site.Relative to prior art, when scanning galvanometer carries out angle scanning, enter
Incidence and outgoing of the P polarization light by keeping parallelism of the first 4F systems are incident upon, no matter how the angle of incident light changes, and is irradiated to
The facula position formed in sensing membrane remains constant, and the 2nd 4F systems of addition in reflected light path, it is ensured that in difference
Angle under, the image space of receiver is not moved, and stability is good and image quality high, so as to ensure in scanning process
SPR excites identical all the time and constant with search coverage, by increasing capacitance it is possible to increase the sensitive zones in sensing membrane, and causes sensitive face has good
Good sensing uniformity, can effectively realize quick detection, and meet the requirement of the highly sensitive detection of quick detection process.Enter one
Step, said system also has the advantages that simple in construction, small volume.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is the accompanying drawing used required in technology description to be briefly described, it should be apparent that, drawings in the following description are only this
Some embodiments of invention, for those skilled in the art, on the premise of not paying creative work, can also basis
These accompanying drawings obtain other accompanying drawings.
Fig. 1 is the structural representation of the novel angle modulation SPR imaging systems based on 4F systems in the embodiment of the present invention；
Fig. 2 is the structural representation of light source module in the embodiment of the present invention；
The schematic diagram of SPR angular spectrum curves when Fig. 3 is different refractivity；
Fig. 4 is spr signal handling principle figure in the embodiment of the present invention；
Fig. 5 is the spr signal curve during protein-interacting；
Fig. 6 is the schematic diagram of the SPR images of protein-interacting formation.
Embodiment
To enable goal of the invention, feature, the advantage of the present invention more obvious and understandable, below in conjunction with the present invention
Accompanying drawing in embodiment, the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described reality
It is only a part of embodiment of the invention to apply example, and not all embodiments.Based on the embodiment in the present invention, people in the art
The every other embodiment that member is obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
Due to angle modulated SPR in the prior art, to there is detection speed slow, and sensitivity is low and the technical problem such as stability difference.
In order to solve the above-mentioned technical problem, the present invention proposes a kind of novel angle modulation SPR imagings system based on 4F systems
System, sets the first 4F systems and the 2nd 4F systems within the system, when scanning galvanometer carries out angle scanning, is incident to the first 4F
The P polarization light of system is by the incidence and outgoing of keeping parallelism, and no matter how the angle of incident light changes, and is irradiated to shape in sensing membrane
Into facula position remain constant, and in reflected light path add the 2nd 4F systems, it is ensured that under a different angle, connect
The image space for receiving device is not moved, and stability is good and image quality is high, so as to ensure that SPR is excited and detected in scanning process
Region is identical all the time and constant, by increasing capacitance it is possible to increase the sensitive zones in sensing membrane, and causes sensitive face to have good sensing consistent
Property, quick detection can be effectively realized, and meet the requirement of the highly sensitive detection of quick detection process.Further, above-mentioned system
System also has the advantages that simple in construction, small volume.
Referring to Fig. 1, SPR imaging systems are modulated for the novel angle based on 4F systems in first embodiment of the invention, should
System includes：
Input path, sensing module and reflected light path；
Input path includes：Light source module 1, the polarizer 2, the 4F systems of scanning galvanometer 3 and the first；Wherein, the first 4F systems
Include the first lens 4 and the second lens 5；
Sensing module includes prism 6, sensing membrane 7 and flow cell 8, and sensing membrane 7 is located between flow cell 8 and prism 6, circulation
Pond 8 is used to sample to be tested flowing through sensing membrane 7；
Reflected light path includes the 2nd 4F systems and receiver 12；Wherein, the 2nd 4F systems are saturating including the 3rd lens 9 and the 4th
Mirror 11.
Further, above-mentioned system also includes analyzer 10, and analyzer 10 is located between the 3rd lens and the 4th lens,
The influence come for the S-polarization light for suppressing to produce in scattering process, the S-polarization light belt produced with removing scattering.
Wherein, light source module 1 is wavelengthtunable light source module, can send the collimated light of different wave length to the polarizer 2, make
Various samples to be tested must can be selected with corresponding output wavelength, realization is most preferably excited.
As described in Figure 2, it is the structural representation of light source module 1, the light source module 1 includes wideband light source 1-1 and wave filter
1-2；Device 1-2 is filtered after filtering after wideband light source 1-1 transmitting light waves, obtains collimated light and the incident polarizer 2, the collimation
Light can be arrowband collimated light.Wherein, wideband light source 1-1 is Coherent Wideband light source, non-coherent broad band light source or part phase
Dry wideband light source, wave filter 1-2 is liquid crystal filter, acousto-optic filter or the wave filter being made up of multigroup optical filter.Or
Person, the light source module 1 can also be made up of the light source of at least two independent different wave lengths.The LD light sources of such as different wave length,
LED/light source etc..
Wherein, the polarizer 2 is used to be filtered the collimated light that light source module 1 is exported, and obtains P polarization light.
Wherein, receiver 12 can be detector array, can be specifically electronic CCD or COMS detectors or array
Photodetector, for collecting reflected light light intensity.
Wherein, flow cell 8 and sensing membrane 7 constitute closed chamber, and testing sample is from the entrance (Sample In) of flow cell 8
Flow into, outlet (Sample Out) outflow, the surface of the sensing membrane 7 under flow cell 8 is flowed through, while to be measured point of testing sample
Son and probe molecule association reaction, make sensing membrane near surface refractive index change, incident P polarization light is on the surface of prism 6
Producing evanescent wave excites free electron in metal film in sensing membrane 7 to produce plasma wave and produce RESONANCE ABSORPTION, is to exist
Sensing membrane 7 excites SPR phenomenons.Wherein, when the near surface refractive index of sensing membrane 7 changes, resonance angle will also occur therewith
Change, the angle change can be obtained by obtaining SPR angular spectrums curve, enabling treated using SPR resonance angles change
The information such as the concentration of test sample product.
Referring to Fig. 3, the schematic diagram of SPR angular spectrum curves during different refractivity, wherein, the variations in refractive index difference of sample
For 1.333,1.336,1.339, its corresponding resonance angle is respectively θ1,θ2,θ3.From the figure 3, it may be seen that when the refractive index of sample occurs
During change, resonance angle will also change, by the change for monitoring resonance angle, it is possible to monitor sample refractive index change or
The situation of person's interaction of molecules, obtains the relevant information of sample.
It is understood that by setting the first 4F systems respectively in input path and reflected light path in present invention implementation
With the 2nd 4F systems, it can remain that directional light is incident to the sensitive face of sensing membrane 7, and the sensing is reflect off with directional light
Face, while into erected image, and under identical clear aperture, scanning angle can reach 2 times of 2F systems, with angle point
Resolution is high, the advantages of sensitivity is high.
SPR imaging systems will be modulated based on the above-mentioned novel angle based on 4F systems below to introduce within the system, light
The transmission path and imaging process on road are as follows：
Light source module 1 sends collimated light by the polarizer 2, forms P polarization light, P polarization light, which is incident to scanning galvanometer 3, to be occurred
Reflection, wherein, the scanning galvanometer 3 is located at the front focal plane of the first lens 4 of the first 4F systems, and the scanning galvanometer 3 is according to default
Step-length rotated, with the incident direction of the P polarization light of the first lens 4 for changing the first 4F systems that are incident to, wherein, it is incident
Scope to the incident direction of the P polarization light of the first lens 4 is [- θ ,+θ], and step-length is △ θ.
The P polarization light of incidence angles degree is incident to the first lens 4 in the first 4F systems, and from the outgoing of the second lens 5
Coupled by prism 6, and incide the sensing membrane 7 on the surface of prism 6 and excite SPR phenomenons.
Wherein, sensing membrane 7 is located at the front focal plane of the 3rd lens 9 in the 2nd 4F systems, and receiver 12 is located at the 2nd 4F systems
Reflected P polarized light, the P polarization light of reflection are incident to the 2nd 4F systems by the back focal plane of the 4th lens 11 in system, sensing membrane 7
3rd lens 9, are collected by the 3rd lens 9, because analyzer 10 is located between the 3rd lens 9 and the 4th lens 11, analyzer 10
S-polarization light will be suppressed, interference of the S-polarization light to system is eliminated, signal to noise ratio is improved, suppress the P polarization light after S-polarization light again by the
Four lens 11 are coupled to receiver 12, and the SPR angular spectrum curves of all sensing sites are formed by receiver 12, and feed back to terminal and set
Standby, terminal device generates SPR images using SPR angular spectrums curve.
Wherein, scanning galvanometer 3 often changes the incident direction for the P polarization light for being once incident to the first 4F systems, the P polarization light
Just change once in the incident angle of sensing membrane 7, the intensity of reflected light image of the one width sensing membrane 7 of record of receiver 12, and due to
The regular change of scanning galvanometer 3 is incident to the incident direction of the P polarization light of the first 4F systems, and P polarization light enters sensing membrane 7
Firing angle degree is also by regular change, a series of corresponding regular intensity of reflected light figure for recording sensing membranes 7 of receiver 12
The intensity of reflected light image of picture, i.e. one width sensing membrane 7 of each incident angle correspondence.The intensity of reflected light figure of sensing membrane 7 in theory
Each pixel of picture correspond to a sensing site in sensing membrane, and the intensity level of the pixel of the same position of image
The SPR angular spectrum curves of curve, the i.e. position that an intensity changes with incident angle are constituted with corresponding incident angle, due to connecing
It is two-dimensional detector to receive device 12, therefore, will form a series of intensity with incident angle change curve, i.e., all sensing sites
SPR angular spectrum curves.
By the above-mentioned means, scanning galvanometer 3 often completes single pass, by the parallel whole sensitive zones for obtaining sensing membrane 7
Whole sensing sites SPR angular spectrum curves, and the SPR angular spectrum curves of acquisition are sent to terminal device, terminal by receiver 12
Equipment utilization software recognizes effective minimum value of the resonance angle of every SPR angular spectrum curve, the i.e. intensity of curve, by these resonance angles
As the gray value of image, SPR resonance angle images are formed, and pass through the SPR resonance angles in scanning process or after the end of scan
Image compares computing with benchmark SPR resonance angle images, and its difference is constituted into SPR images.
In order to be better understood from the technical scheme in the embodiment of the present invention, terminal device is described more fully below obtaining
Have after the SPR angular spectrum curves of sensing site, the process of SPR images is obtained by software processing, it is as follows：
1) scanning wavelength determines and sets the output wavelength of light source module 1.The determination of the output wavelength of the light source module 1 can
To be accomplished by the following way：By carrying out spectral scan to incident light source, it is possible to use liquid crystal tunable filter, Acousto-optic filtering
Output light wavelength and gray-scale relation curve, i.e. SPR spectroscopy curve at the means such as device or optical filter runner, record target point, pass through
Fitting, interpolation scheduling algorithm processing, obtain its corresponding resonant wavelength, using the resonant wavelength as light source module 1 output wavelength.
2) based on the system in above-described embodiment, incident angle is scanned using the scanning galvanometer 3 in the system, and
By the corresponding intensity of reflected light image of the synchronous recording of receiver 12, the gray level image of sensitive face under one group of different angle is obtained.
3) gray value for extracting corresponding pixel points from this group of gray level image obtained constitutes angle-intensity curve, i.e. SPR angles
Spectral curve.Because receiver 12 is two-dimentional, in theory, there is one-to-one relationship in sensing site, therefore with receiver pixel
Each gray value of the pixel in this group of gray level image can constitute a SPR angular spectrum curve, so a series of SPR angles can be constituted
Spectral curve.If for example, this group of gray level image has 100 width, and for pixel i, pixel i is in the 100 width gray level image
All there is gray value, 100 gray values are had, because each width gray level image all corresponds to an angle, then by 100 gray scales
Value and the corresponding angle of each gray value just can structure at an angle-intensity curve, i.e. SPR angular spectrums curve.Certainly, we
Area pixel can also be carried out to a series of this gray level image to be averaged, constitute average gray image, be conducive to reducing system noise.
If for example, each image in this group of gray level image has 1024*1024 pixel, and having 100 width gray level images, then can
Being formed on 1024 SPR angular spectrum curves, every curve has 100 gray values, and the gray level image is compressed into picture by zone leveling
Element, i.e., be compressed into 256*256 by 1024*1024, then can obtain 256 SPR angular spectrum curves, every 100 gray values.
4) the SPR angular spectrum curves of acquisition are fitted, the Mathematical treatment such as difference, improve angular resolution, and extract
The SPR resonance angles of corresponding angle value, the i.e. corresponding sensing site of the pixel, every song at the intensity minimum of SPR angular spectrum curves
Line is all to that should have respective SPR resonance angles, further, also using all curves SPR resonance angles as image gray scale
Value, forms SPR resonance angle images.
5) above-mentioned step 2 is repeated to 4, the resonance angle of all sensing sites in the same time is obtained not, forms resonance angle at any time
Between the curve that changes, the SPR resonance angles change of each sensing site can also be converted into gray value, form SPR images.
Referring to Fig. 4, being the spr signal handling principle figure in the embodiment of the present invention, scanning galvanometer 3 often changes once incident
As soon as the incident angle of the P polarization light to 4F systems, the whole sensitive zones progress to sensing membrane 7 is Polaroid, work as incidence
Angle is θNWhen, correspondence collection N width images, the gray value of each pixel represents the intensity of the reflected light of the pixel record
Value, when angle is from θ1To θNWhen at the uniform velocity changing, angle-intensity curve of each pixel, i.e. SPR angular spectrums curve, and can be obtained
SPR resonance angles are obtained using SPR angular spectrum curves, and reconstruct the SPR resonance angle images of two dimension.The SPR of any time is resonated
Angle image is compared with baseline SPR resonance angle images, you can obtain SPR images.
In embodiments of the present invention, by setting the first 4F systems and the 2nd 4F systems to be combined with scanning galvanometer so that energy
Enough quick progress SPR resonance angles diagnosis are simultaneously imaged, while with high sensitivity, high uniformity and quick detection molecules phase interaction
With so as to which the requirement to sensing membrane is greatly lowered.
Referring to Fig. 5, being the SPR angular spectrum curves during protein-interacting, Fig. 6 is protein-interacting formation
The schematic diagram of SPR images, gray value represents resonance angle variable quantity before and after reaction.
In the above-described embodiments, the description to each embodiment all emphasizes particularly on different fields, and does not have the portion being described in detail in some embodiment
Point, it may refer to the associated description of other embodiments.
It is the description that SPR imaging systems are modulated to a kind of novel angle based on 4F systems provided by the present invention above,
For those skilled in the art, according to the thought of the embodiment of the present invention, have in specific embodiments and applications
Change part, to sum up, this specification content should not be construed as limiting the invention.
Claims (10)
1. a kind of novel angle modulation SPR imaging systems based on 4F systems, it is characterised in that including input path, sensing mould
Block and reflected light path；
The input path includes：Light source module, the polarizer, scanning galvanometer and the first 4F systems；
The sensing module includes prism, sensing membrane and flow cell, and the flow cell is used to sample to be tested flowing through the biography
Feel film；
The reflected light path includes the 2nd 4F systems and receiver；
The light source module sends collimated light by the polarizer, forms P polarization light, and the P polarization light is incident to described sweep
Retouch galvanometer to reflect, the P polarization light of reflection is incident to the first 4F systems, through described after the first 4F system exits
Prism-coupled, and incide the sensing membrane of prism surface and excite SPR phenomenons, the sensing membrane is by reflected P polarized light, the P of reflection
Polarised light is incident to the 2nd 4F systems, by the 2nd 4F system exits to the receiver, is formed by the receiver
The SPR angular spectrum curves of all sensing sites, and terminal device is fed back to, the terminal device is given birth to using the SPR angular spectrums curve
Into SPR images.
2. system according to claim 1, it is characterised in that the first 4F systems are saturating comprising the first lens and second
Mirror, the P polarization light of the scanning galvanometer reflection is projected from first lens entrance and from second lens.
3. system according to claim 1, it is characterised in that the 2nd 4F systems are saturating comprising the 3rd lens and the 4th
Mirror, and the sensing membrane reflection P polarization light from the 3rd lens entrance and from the 4th lens project.
4. system according to claim 3, it is characterised in that the system also includes analyzer, the analyzer is located at
Between 3rd lens and the 4th lens, for the S-polarization light for suppressing to produce in scattering process.
5. system according to claim 1, it is characterised in that the scanning galvanometer is rotated according to preset step-length, to change
Become the incident direction for the P polarization light for being incident to the first 4F systems, the scope of the incident direction is [- θ ,+θ].
6. system according to claim 1, it is characterised in that the receiver is detector array.
7. system according to claim 6, it is characterised in that the detector array is electronic CCD or cmos detector
Or array photodetectors.
8. system according to claim 1, it is characterised in that the light source module is wavelengthtunable light source module.
9. system according to claim 8, it is characterised in that the light source module includes wideband light source and wave filter；Institute
State and pass through the filter filtering after wideband light source launching beam, obtain the collimated light and the incident polarizer；
The wideband light source is Coherent Wideband light source, non-coherent broad band light source or partially coherent wideband light source, the wave filter
The wave filter constituted for liquid crystal filter, acousto-optic filter or by multigroup optical filter.
10. system according to claim 8, it is characterised in that the light source module is by least two independent different ripples
Long light source is constituted.
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| CN109444078A (en) * | 2019-01-14 | 2019-03-08 | 兰州理工大学 | Length scanning index sensor based on unsymmetrical metal cladding Medium Wave Guide |
| CN112098371A (en) * | 2020-09-15 | 2020-12-18 | 深圳大学 | Strength type SPRi sensing system and method based on dual-wavelength difference |
| CN113376225A (en) * | 2021-01-20 | 2021-09-10 | 中国科学技术大学 | High-resolution imaging device for nano material and imaging analysis method thereof |
| TWI797631B (en) * | 2021-05-28 | 2023-04-01 | 國立成功大學 | Method for detecting the object by changing the refractive index in the metal surface plasma principle (spp) |
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