CN104237170A - Surface plasma resonance imaging sensor detection system - Google Patents
Surface plasma resonance imaging sensor detection system Download PDFInfo
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- CN104237170A CN104237170A CN201410531657.0A CN201410531657A CN104237170A CN 104237170 A CN104237170 A CN 104237170A CN 201410531657 A CN201410531657 A CN 201410531657A CN 104237170 A CN104237170 A CN 104237170A
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Abstract
The invention discloses a surface plasma resonance imaging sensor detection system which comprises a light source. Light emitted by the light source passes through a Galileo light beam expander and is diffused into even light beams, then the light beams sequentially pass through a polarization apparatus and a variable raster, the light is irradiated onto the sensing chip surface on a circulating cell, finally the light is reflected by a prism and chips and fed into a CCD imaging system, and a corresponding sensing curve and a real-time imaging result are output through a computer. In the surface plasma resonance imaging sensor detection system, a surface plasma resonance imaging technology (SPRi) is a novel mark-free and high-flux optical sensing technology integrated with an SPR technology, CCD imaging and arraying chips, and the surface plasma resonance imaging sensor detection system is widely applied to mutual effect study of bio-molecules.
Description
Technical field
The present invention relates to a kind of surface plasmon resonance imaging technique, particularly relate to a kind of surface plasma resonance image-forming sensing and detecting system.
Background technology
Surface plasma resonance imaging (surface plasmon resonance imaging, SPRi) technology is based on surface plasma resonance (surface plasmon resonance, SPR) technology, in conjunction with a kind of optical image technology that CCD imaging and array chip grow up.Surface plasma resonance (SPR) is the electric density vibration resonance ripple be present on metal and dielectric interface, can excite by incident electromagnetic wave.When incident light from optically denser medium propagate into optically thinner medium and incident angle in suitable scope time, in the interface experiences total internal reflection of two kinds of materials.SPR is very responsive to the subtle change of interfacial medium refractive index, and when sample and interracial contact, owing to there is absorption or chemical reaction, interface medium refraction index will change.Based on this principle can real-time, the monitoring bio molecule that can't harm of original position and between interaction, the dynamic process studied biological intermolecular combination and dissociate, understands the physics and chemistry adsorption process at interface.As the strong biomolecule analysis instrument of one, surface plasmon resonance biosensor possesses in real time, fast, exempt from mark, favorable reproducibility, highly sensitive, sample without the need to advantages such as purifying, has been widely used in protein, nucleic acid, and micromolecular detection analysis.And surface plasma resonance imaging technology is while possessing surface plasma resonance technology advantage, rely on again the combination of SPRi technology and array biochip technology, by probe arrays different for bio-sensing chip finishing, realize the high-throughout detection to testing sample target substances and screening.In recent years, for adapting to the arrival of " genome times afterwards comprehensively ", the research of the various molecular biosciences groups such as genomics, proteomics and metabolism group deepens continuously, surface plasma resonance imaging technology is more and more subject to extensive concern and the attention of numerous researcher, is considered to the very potential high flux bioanalytical method of one.
Summary of the invention
In view of the demand of prior art, the object of the present invention is to provide a kind of surface plasma resonance image-forming sensing and detecting system.
A kind of surface plasma resonance image-forming sensing and detecting system, comprise light source, the light that light source sends is subsequently by Galileo beam-expander, the light sent by pointolite diffuses into uniform light beam, light beam is then successively by polarizer, iris shutter, then light is irradiated to the censorchip surface on flow cell, and last light, by the reflection of prism and chip, enters CCD imaging system and then exports corresponding sensing curve and real time imagery result by computer.
As preferably, described flow cell, comprise base, described base is provided with flow cell main part, described flow cell main part inner transverse is provided with infiltration block, described infiltration block is the airtight tubular in one end, airtight by end cap in the flow cell main body of corresponding described infiltration block opening end, end cap is provided with at least two openings, the sealed end end face of described infiltration block is provided with aperture, the circulation main component of corresponding described infiltration block sealed end is provided with gasket recess, described gasket recess is embedded in packing ring, fit with chip outside described packing ring and form working cavity, described chip is compressed by prism, described prism is compressed by flow cell annex, described flow cell annex correspondence is arranged on the rear of flow cell main part, described flow cell annex comprises the holder be fastened on base, compact heap is provided with between described holder and described prism, on described holder, horizontally set adjusting screw is for compressing compact heap.
As preferably, described chip through gold-plated treatment process, forms the dot matrix chip of array by microslide.
As preferably, described compact heap side is provided with the groove running through upper and lower surface, and described groove fixes the seamed edge of prism.
As preferably, under described flow cell, chain is connected to a universal stage (being mainly used in the adjustment of the angle of flow cell entirety) of a horizontal slope regulating platform (being mainly used in regulating height and the horizontality of flow cell) and bottom.
As preferably, described flow cell is also connected with sampling system, mainly comprises six-way valve and precise micro syringe pump.
As preferably, described optical source wavelength is the LED of 650nm, is connected on small-sized DC power supply, by the linear voltage stabilization mode of low ripple low noise, light source voltage is stablized, ensures stable light intensity.
As preferably, described light source is fixed by threaded base, this threaded base is fixed on adjustable pin hole base, therefore can carry out accurate adjustment to the XY direction of light source, ensure the accurate of light path.
As preferably, described polarizer is fixed on polarizer universal stage, can regulate polarizer direction, can reach control different directions polarized light by the function of polarizer.
As preferably, the base of flow cell, main part and annex are made up of hard aluminium alloy.
As preferably, compact heap side is provided with the groove running through upper and lower surface, and described groove fixes the seamed edge of prism.
As preferably, infiltration block is processed by polymethylmethacrylate.
As preferably, chip through gold-plated treatment process, forms the dot matrix chip of array by microslide.
As preferably, packing ring is processed by teflon.
As preferably, prism is made up of flint glass.
As mentioned above, according to surface plasma resonance image-forming sensing and detecting system of the present invention, there is following beneficial effect:
Surface plasma resonance imaging technology (SPRi) be integrate SPR technique, the Novel free of CCD imaging and array chip marks, high-throughout optical sensing technology, is widely used in the repercussion study of biomolecule.SPRi possesses in real time, fast, exempt from mark, favorable reproducibility and sensitivity advantages of higher, and energy mimic biology environment, carries out high-throughout biochemical analysis, especially in monitoring interaction of molecules and performance analysis, have huge advantage.
Accompanying drawing explanation
Fig. 1 is shown as the structural representation of detection system of the invention process.
Fig. 2 is shown as the structural representation I of the flow cell of the embodiment of the present invention.
Fig. 3 is shown as the structural representation II of the flow cell of the embodiment of the present invention.
Fig. 4 is shown as the flow cell side sectional view of the embodiment of the present invention.
Fig. 5 is shown as the spectrogram adopting the detection system of the present embodiment to obtain.
Piece mark explanation
1 base,
2 circulation main parts,
3 infiltration blocks,
4 circular end caps,
5 packing rings,
6 chips,
7 prisms,
8 compact heaps,
9 flow cell annexes,
10 adjusting screws.
Embodiment
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
The present embodiment provides a kind of surface plasma resonance image-forming sensing and detecting system, first choose wavelength be the LED of 650nm as light source, be connected on small-sized DC power supply, by the linear voltage stabilization mode of low ripple low noise, light source voltage is stablized, ensures stable light intensity; Then fix light source by threaded base, this threaded base is fixed on adjustable pin hole base, therefore can carry out accurate adjustment to the XY direction of light source, ensure the accurate of light path; The light that light source sends is subsequently by Galileo beam-expander, the light sent by pointolite diffuses into uniform light beam, all parallel and the energy of light beam is the same, there is no deviation, then polarizer is passed through, polarizer is fixed on polarizer universal stage, can regulate polarizer direction, can reach the function of polarized light by polarizer of control different directions; Then light can pass through an iris shutter, can be controlled the number of incident ray by adjuster bar; Then light is irradiated to the censorchip surface on flow cell, and under flow cell, chain is connected to one for the horizontal slope regulating platform of height and horizontality that regulates flow cell and a universal stage for the angular adjustment of flow cell entirety of bottom; Last light, by the reflection of prism and chip, enters CCD imaging system and then exports corresponding sensing curve and real time imagery result on computers by software kit.
' on small-sized DC power supply of the present embodiment employing, linear voltage stabilization mode by low ripple low noise ', it is low to there is transformer conversion efficiency in conventional linear stabilized voltage supply, voltage stabilizing chip pressure reduction is large, the shortcomings such as filtering circuit is perfect not, often occur that output ripple is large, thermal value is large, efficiency is low, indirectly increase the problems such as thermonoise to system, and on this small-sized DC power supply, by voltage reduction module, rectification filtering module, linear voltage stabilization module and low-pass filtering module composition, therefore power supply not only has high stability, the advantage of low ripple, and output voltage fluctuation is little, voltage is adjustable, carrying load ability is strong, compact.
The threaded base that the present embodiment adopts, mainly according to adjustable pin hole base, is mainly used in the adjustment of the fixing of light source and position.
' the iris shutter that the present embodiment adopts, the number of incident ray can be controlled by adjuster bar ', on iris shutter, there is an adjuster bar, by the size of the adjustment adjustable variable grating central aperture of adjuster bar, therefore can control the amount of the light by this iris shutter and regulate.
' for the horizontal slope regulating platform of height and horizontality that regulates flow cell and a universal stage for the angular adjustment of flow cell entirety of bottom ' that the present embodiment adopts, the top of horizontal slope regulating platform is fixed with flow cell, prism and chip, by the adjustment of this regulating platform, the central point of flow cell, prism and chip can be made to be positioned at the central point of whole platform, and remain on a level height with incident ray, be conducive to subsequent experimental.And the universal stage of bottom mainly carries out rotation adjustment by extension bar connected system to the prism of top, thus regulate the angle of incident ray, carry out the adjustment of different refractivity solution best angle.
' software kit ' software that the present embodiment adopts is based on up-to-date Labview, and the function experimentally needed for platform, design and programme corresponding operating function, realizes easy and simple to handle, multiple functional control software design.
The flow cell that the present embodiment adopts, comprise base 1, base 1 is square stereo block, and the side above base 1 is provided with flow cell main part 2, and circulation main part 2 is a stereo block upwards extended, integrally manufactured with base 1.The intermediate lateral of flow cell main body 2 arranges a circular through hole, settles infiltration block 3 in through hole.Infiltration block 3 is the airtight cylinder in an one end, forms circulation cavity, and the liquid communication volume in circulation cavity is at 30 ~ 50ul.The flow cell of corresponding infiltration block 3 openend fixedly mounts a circular end cap 4 by screw thread, and circular end cap 4 is distributed with two openings, in the present embodiment, the opening of below is injection port, and the opening of top is outlet.All openings are connected with external sample delivery pipeline.The sealed end end face of infiltration block 3 is provided with aperture, and this aperture passes for the testing liquid circulated in cavity.The circulation main body 2 of corresponding infiltration block 3 sealed end is provided with gasket recess, and gasket recess is embedded in packing ring 5, and the packing ring 5 after embedding is a little more than the back flat of flow cell main body 2.The flow cell of the present embodiment also comprises chip 6, chip 6 packing washer 5, makes chip 6, packing ring 5 forms a reaction chamber.The equilateral prism 7 that chip 6 is 25mm by a length of side compresses.A vertical plane of prism 7 is close to chip, and the seamed edge that binding face is relative is compressed by compact heap 8.Compact heap 8 is a three-dimensional bulk, and vertically arranges groove, seamed edge and a groove fit.
The back side of corresponding flow cell main part 2 is provided with flow cell annex 9.Flow cell annex 9 is a holder being fixed on base, and holder both sides are provided with step, and step is provided with mounting hole, is connected with base 1 by mounting hole mounting screw.Holder is horizontally arranged with two mounting holes, in mounting hole, penetrates the adjusting screw 10 equal with aperture, prop up compact heap 8 by adjusting screw 10 end, compress triangular prism 7 and chip 6.
The base 1 of this flow cell of the present embodiment, main part 2 and annex 9 are made up of hard aluminium alloy.
The infiltration block 3 of the present embodiment processes with polymethylmethacrylate.Use the infiltration block 3 of this material not easily broken, there is the transparency of height, be easy to observe internal liquid state and whether have bubble to disturb; And there is chemical-resistant reagent and solvent resistance.
The chip 6 of the present embodiment through gold-plated treatment process, forms the dot matrix chip of array by microslide.Can be used for high throughput analysis application.Gold-plated treatment process be by microslide after the cleaning treatment of Piranha solution and acid solution, rinse with deionized water again, 70 DEG C of thermal treatments one hour is re-used after drying up with nitrogen subsequently, finally carry out electron beam evaporation (electron beam evaporation) process, the microslide handled well plates the Cr that one deck 2nm is thick, and then plates the thick gold of one deck 51nm.The dot matrix chip of the present embodiment, it is exactly array chip, namely when making chip, experimentally, time deposited by electron beam evaporation technology carries out plated film, optionally carry out 4 × 4,8 × 8, the point of the array of 10 × 10 grades, each point is the elementary cell of a reaction monitoring, therefore can carry out high-throughout detection.
The packing ring 6 of the present embodiment is processed by teflon.There is fabulous thermotolerance, moisture resistance, mar proof and corrosion resistance, be suitable for the various conventional buffers of different detection system.Its external diameter 19.05mm, internal diameter 15.88mm.
The equilateral prism of the prism 7 of the present embodiment to be the length of side be 25mm, be made up of F2 glass material, F2 is a kind of flint glass, good performance is had in visible ray and near infrared spectral range, there is high reflectance and low Abbe number, and compared with N-SF11, it has higher chemical resistance and the transmissivity of Lve Gao.
The flow cell of the present embodiment connects six-way valve by the injection port on end cap, then by precise micro syringe pump energy supply sample detection, then connected outside by the outlet on end cap, get rid of waste material.Testing liquid, by injection port, flows through the hole of mainstream channel again on infiltration block of infiltration block in circulation cavity, reaches the confined space that the flow cell other end is formed by prism, chip and packing ring.Liquid flows through chip surface in this space, the material that test substance in liquid and chip surface have been modified fully contacts and reacts, realize interactional process, then under the promotion of damping fluid, this part testing liquid flows to outlet from mainstream channel again subsequently, be discharged to waste liquid cylinder, whole process just completes once.This flow cell have employed base and the annex of hard aluminium alloy formation, and with polymethylmethacrylate process have circulation cavity Liquid Penetrant block, form the system for the distribution of commodities of precise micro, a small amount of testing sample can be detected, and the high throughput analysis of many materials can be realized in conjunction with array chip.
The present embodiment is applicable in concrete experiment:
First by chip anthropophagy's fishing solution (dense H
2sO
4: H
2o
2=7:3) clean, then rinse well with anthropophagy's fishing solution of distilled water by chip surface, with nitrogen, chip is dried up; Drip matching fluid (matching fluid) to two at the chip surface of drying, then chip, prism and flow cell are fitted together; The flow velocity arranging precise micro syringe pump is 1ml/min, installs deionized water in advance in precision syringe, as the operation liquid (running buffer) of whole experiment; Opening the software design patterns time shutter is 4ms, choose region to be detected, then syringe pump is opened, after band baseline stability, six-way valve is modulated Load state, inject the glucose solution of variable concentrations wherein, after allowing, six-way valve is modulated Inject state, while testing liquid flows through chip surface, observe the change of performance graph in control software design interface, the drafting that the data of having preserved carry out curve can be opened after stopping experiment.With reference to the accompanying drawings shown in 5, the glucose solution of what each peak was corresponding is variable concentrations, because the refractive index of the glucose solution of variable concentrations is different, therefore can cause the difference of reflective light intensity, namely the difference in height at peak shown in figure.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all without prejudice under spirit of the present invention and category, can modify above-described embodiment or changes.Therefore, such as have in art usually know the knowledgeable do not depart from complete under disclosed spirit and technological thought all equivalence modify or change, must be contained by claim of the present invention.
Claims (9)
1. a surface plasma resonance image-forming sensing and detecting system, it is characterized in that, comprise light source, the light that light source sends is subsequently by Galileo beam-expander, the light sent by pointolite diffuses into uniform light beam, and light beam then successively by polarizer, iris shutter, then is irradiated to the censorchip surface on flow cell, last light, by the reflection of prism and chip, enters CCD imaging system and then exports corresponding sensing curve and real time imagery result by computer.
2. detection system according to claim 1, it is characterized in that, described flow cell, comprise base, described base is provided with flow cell main part, described flow cell main part inner transverse is provided with infiltration block, described infiltration block is the airtight tubular in one end, airtight by end cap in the flow cell main body of corresponding described infiltration block opening end, end cap is provided with at least two openings, the sealed end end face of described infiltration block is provided with aperture, the circulation main component of corresponding described infiltration block sealed end is provided with gasket recess, described gasket recess is embedded in packing ring, fit with chip outside described packing ring and form working cavity, described chip is compressed by prism, described prism is compressed by flow cell annex, described flow cell annex correspondence is arranged on the rear of flow cell main part, described flow cell annex comprises the holder be fastened on base, compact heap is provided with between described holder and described prism, on described holder, horizontally set adjusting screw is for compressing compact heap.
3. detection system according to claim 2, is characterized in that, described chip through gold-plated treatment process, forms the dot matrix chip of array by microslide.
4. detection system according to claim 3, is characterized in that, described compact heap side is provided with the groove running through upper and lower surface, and described groove fixes the seamed edge of prism.
5. detection system according to claim 2, it is characterized in that, link the horizontal slope regulating platform for the height and horizontality regulating flow cell below described flow cell, the bottom of described horizontal slope regulating platform is provided with the universal stage for regulating flow cell angle.
6. detection system according to claim 2, is characterized in that, described flow cell is connected with sampling system, and described sampling system comprises six-way valve and precise micro syringe pump, and described six-way valve connects the opening on flow cell end cap.
7. detection system according to claim 1, is characterized in that, described light source is the LED of wavelength 650nm, described LED is connected on small-sized DC power supply, by the linear voltage stabilization mode of low ripple low noise, light source voltage is stablized, ensure stable light intensity.
8. detection system according to claim 1, is characterized in that, described light source is fixed by threaded base, this threaded base is fixed on adjustable pin hole base, therefore can carry out accurate adjustment to the XY direction of light source, ensure the accurate of light path.
9. detection system according to claim 1, is characterized in that, described polarizer is fixed on polarizer universal stage, can regulate polarizer direction, can reach the function of polarized light by polarizer of control different directions.
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| CN110261350A (en) * | 2019-06-04 | 2019-09-20 | 暨南大学 | A kind of optical sensor system and sensing testing method |
| CN111380839A (en) * | 2020-04-27 | 2020-07-07 | 上海工程技术大学 | SPR sensor device based on liquid bag angle adjustment |
| CN112858223A (en) * | 2021-01-11 | 2021-05-28 | 深圳大学 | SPR sensing device and method based on nano photo-thermal tweezers technology |
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| CN112858223A (en) * | 2021-01-11 | 2021-05-28 | 深圳大学 | SPR sensing device and method based on nano photo-thermal tweezers technology |
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Effective date of registration: 20220719 Address after: 400030 Building 5, Lanyuan, No. 61, Middle University Town Road, Shapingba District, Chongqing Patentee after: International Institute of in vitro diagnostics, Chongqing Medical University Address before: 400016 No. 1, Medical College Road, Yuzhong District, Chongqing Patentee before: Chongqing Medical University |