WO2009078510A1

WO2009078510A1 - Disposable surface plasmon resonance biosensor and system using the same

Info

Publication number: WO2009078510A1
Application number: PCT/KR2008/003113
Authority: WO
Inventors: Hyeon Bong Pyo; Moon Youn Jung; Seon Hee Park
Original assignee: Electronics and Telecommunications Research Institute ETRI
Current assignee: Electronics and Telecommunications Research Institute ETRI
Priority date: 2007-12-17
Filing date: 2008-06-04
Publication date: 2009-06-25
Anticipated expiration: 2010-06-17
Also published as: DE112008003435T5; KR100922578B1; KR20090064916A

Abstract

A Surface Plasmon Resonance (SPR) biosensor and system using the same are provided. The biosensor includes a sensor substrate and a thin metal film. The sensor substrate is configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body. The thin metal film is formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed and generates SPR by light impinged on the prism.

Description

DISPOSABLE SURFACE PLASMON RESONANCE BIOSENSOR

AND SYSTEM USING THE SAME

Technical Field

[1] The present invention relates to a biosensor for quantitatively measuring interactions between biomolecules in real time using Surface Plasmon Resonance (SPR) and a system using the same.

[2] The invention has been supported by ITR&D Program of MIC/IITA [2006-S-007-02,

Ubiquitous Health Monitoring Module and System Development]. Background Art

[3] If light polarized in a Transverse Magnetic (TM) mode is incident on a thin metal film such as aurum (Au) and argentums (Ag) the real part of a dielectric function of which has a negative (-) value, the light is coupled with surface Plasmon and a resonance phenomenon occurs at a specific angle of incidence satisfying a resonance condition. This is called 'Surface Plasmon Resonance (SPR)'. The surface plasmon is a collective oscillation mode of electrons at a metal surface. Particularly, in a resonance condition where a wave vector of light incident on a thin metal film matches with that of surface plasmons, the intensity of light undergoing the total reflection at a metal surface is consequently minimized because the energy of the incident light is almost all absorbed in a surface plasmon mode.

[4] The resonance condition changes due to the change of the refractive index of dielectric material on a metal surface. By measuring this change, physical, biophysical- as well as biochemical interactions of molecules can be quantitatively analyzed in real time.

[5] Efforts to apply this SPR phenomenon to bio- or chemical sensors have been made continuously. As a result, SPR-based biosensors have become one of the typical non- labeling sensor systems that are able to measure biomolecular interactions without any labels such as fluorophores.

[6] A variety of SPR-based biosensor systems such as angle-, wavelength-, and intensity-interrogated SPR sensors have been developed up to now. For example, in a wavelength-interrogated SPR sensor system using a polarization-maintaining glass fiber, in which a TM-polarized, polychromatic light source is launched, the guided light wave is totally reflected at the dielectric (partially exposed core) and ambient (e.g. air, water, etc.) interface. If a thin noble metal film is formed on top of exposed fiber core surface, the changes in (effective) optical thickness or refractive index on metal layer results in the SPR wavelength change, which can be observed via a spec- trometer. In one another type of the SPR sensor, the miniaturization is accomplished by integrating a light source and a light receiving unit into a plastic light waveguide.

[7] FIGS. 1 to 3 are schematic diagrams illustrating the constructions of conventional

SPR biosensors. Referring to FIGS. 1 to 3, the conventional SPR biosensors are based on so called Kretschmann-Raether configuration to pass an incident light through a prism 10, which is made of a transparent dielectric material with a high refractive index, and to reflect light internally in order to increase a wave vector or momentum of light and to couple it to the surface plasmons.

[8] In the SPR biosensor of FIG. 1, an incident light of monochromatic wave is generated from a light source 30, polarized through a polarizer 31, and then incident on a prism 10. By moving the light source 30 using a goniometer (not shown) to vary an angle of incidence, the SPR biosensor measures, by monitoring the SPR angle change, the change of effective refractive index or of effective thickness change, caused by the existence of a dielectric material 40 adsorbed on a thin metal film 20.

[9] In the SPR biosensor of FIG. 2, an incident light of monochromatic wave generated from a light source 30 is extended and provided in a two-dimensional plane form with its angle of incidence being kept fixed. By doing so, the SPR biosensor expresses, by a relative brightness difference, the changes in effective refractive index or effective thickness, which results from the existence of a dielectric material 40 as appeared in each points on a thin metal film 20. In general, such a SPR biosensor is applied in a form of a multi channel sensor system.

[10] In the SPR biosensor of FIG. 3, an incident light of monochromatic wave generated from a light source 30 is focused using a lens 33 so that the incident light can be always normal to the surface of a prism 10. Like the SPR biosensor of FIG. 1, the SPR biosensor of FIG. 3 measures, by a change of SPR angle, a change of an effective refractive index or effective thickness caused by a dielectric material 40 on a thin metal film 20.

[11] However, in the conventional SPR biosensors, the thin metal film 20 is deposited not directly on the surface of the prism 10 but onto a flat, transparent dielectric substrate 11 such as a slide glass or a microscope cover slip having the same refractive index as the prism 10 in order to make it easy to form, e.g., a Self-Assembled Monolayer (SAM) on a sensor surface or to facilitate other biochemical processing and also for further use of the prism 10 which is made of high refractive index glass (e.g., BK7, SFI l, LaSFN9, etc.).

[12] Thus, the conventional SPR biosensors require the introduction of a medium for optical coupling between the prism 10 and the flat type transparent dielectric substrate 11. Currently, a fluid such as index-matching oil or a solid, transparent elastomer has been used for this purpose. [13] As described above, in the conventional SPR biosensors, the flat type transparent dielectric substrate 11 and the prism 10 with high refractive index glass material were made separately and therefore, there have been a lot of inconveniencies and difficulties in using a fluid such as index-matching oil.

[14] In detail, it is difficult to avoid contamination of sensor surface because it heavily depends on the degree of user's skill. Also, as time goes, index-matching oil, a fluid generates air bubbles or is easily vaporized, and is generally toxic for high refractive index oil. In particular, in the case of the SPR biosensor of FIG. 2, the formation of air bubbles from the index-matching oil appears as a deterioration of the image quality. Disclosure of Invention Technical Problem

[15] The present invention has been made to solve the foregoing problems with the prior art, and therefore the present invention provides a disposable SPR biosensor being applicable to various types of SPR sensor systems without index-matching oil (or a transparent solid body having the same function as the index-matching oil) between a sensor chip having a thin metal film and a prism, which is to couple the incident light with the surface plasmons of the thin metal surface. Technical Solution

[16] According to an aspect of the present invention, a Surface Plasmon Resonance (SPR) biosensor is provided. The biosensor includes a sensor substrate and a thin metal film. The sensor substrate is configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body. The thin metal film is formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed and generates SPR by light impinged on the prism.

[17] According to another aspect of the present invention, a Surface Plasmon Resonance

(SPR) biosensor system is provided. The system includes an SPR biosensor, a light source, a light receiving unit, a polarizer, and a signal processor. The SPR biosensor includes a sensor substrate and a thin metal film. The sensor substrate is configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body. The thin metal film supports surface plasmon and is formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed. In the SPR biosensor, light is impinged on the prism into the upper surface of the substrate just beneath the thin metal film, and performs the total internal reflection (TIR) above the critical angle. The light source provides the light impinges on the prism. The light receiving unit receives the reflected light, which is emitted through the prism, from the center of the prism and converts the reflected light into an electrical signal. The polarizer decomposes the incident or the reflected light emitted from the prism, in a Transverse Magnetic (TM) mode. The signal processor analyzes the electrical signal from the light receiving unit and detects a change of an effective thickness or effective refractive index of a sample, on top of the thin metal film by monitoring the change of SPR angle.

[18] The biosensor may further include one or more channels which are formed on the thin metal film across the SPR absorption dip line, at which a reflected light is minimized by plasmon resonant absorption of incident light, and all or part of the channels may be formed of different dielectric material.

[19] According to a further another aspect of the present invention, a Surface Plasmon

Resonance (SPR) biosensor system is provided. The system includes an SPR biosensor, a light source, a polarizer, and a two-dimensional imaging unit. The SPR biosensor includes a sensor substrate, a thin metal film, and one or more channels. The sensor substrate is configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body. The thin metal film supports surface plasmon and is formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed. The one or more channels are formed on the thin metal film across the SPR absorption dip line, at which a reflected light is minimized by Plasmon resonant absorption of incident light, and all or part of the channels may be formed of a different dielectric material. In the SPR biosensor, light is impinged on the prism into the upper surface of the substrate just beneath the thin metal film, and performs the total internal reflection (TIR) above the critical angle. The light source provides the light impinges on the prism. The polarizer decomposes the incident or the reflected light emitted from the prism, in a Transverse Magnetic (TM) mode. The two-dimensional imaging unit displays the reflected light emitted through the prism, as a two-dimensional plane image.

[20]

Advantageous Effects

[21] As set forth above, the SPR biosensor of the present invention realizes a prism and a flat type transparent substrate having a thin metal film in a body by injection molding of an optical polymer having a high refractive index. By doing so, the SPR biosensor has excellent effects that it has no need to introduce index-matching oil between a sensor chip and the prism to match the refractive index, can be manufactured relatively easily. In addition, a manufacturing cost is greatly saved compared to a conventional SPR biosensor using a high refractive glass substrate, simplification, miniaturization and lightweighting can be accomplished, and a disposable use can be made.

[22] Also, the SPR biosensor of the present invention has an advantage of, if necessary, including a light source, a light receiving unit, a lens, and an imaging unit and being applicable to diverse types of SPR sensor systems having a different structure, by realizing only a sensor chip and a prism in a body. Brief Description of the Drawings

[23] FIGS. 1 to 3 are schematic diagrams illustrating the constructions of conventional

SPR biosensors;

[24] FIG. 4 is a view illustrating a construction of an SPR biosensor according to the first exemplary embodiment of the present invention;

[25] FIG. 5 is a view illustrating a construction of an SPR biosensor according to the second exemplary embodiment of the present invention;

[26] FIG. 6 is a schematic diagram illustrating the sensing principle using the SPR biosensor according to the second exemplary embodiment of the present invention;

[27] FIG. 7 is a graph showing reflectivity versus angle of incidence in a SPR biosensor according to the first exemplary embodiment of the present invention;

[28] FIG. 8 is a graph showing reflectivity versus angle of incidence in a SPR biosensor according to the second exemplary embodiment of the present invention; and

[29] FIG. 9 is a diagram illustrating a case where biomolecules and target molecules are specifically bound on a SPR biosensor according to the second exemplary embodiment of the present invention. Best Mode for Carrying Out the Invention

[30] Preferred embodiments of the present invention will be described herein below with reference to the accompanying drawings. In the following description, well-known functions or constructions are not described in detail since they would obscure the invention in unnecessary detail.

[31] Throughout the drawings, it should be noted that like reference numbers are used to depict the same or similar elements, features and structures.

[32] Throughout the specification, 'connecting of any part with other part' includes not only 'directly connecting' but also 'indirectly connecting' with another element interposed between them. Including any element' signifies not excluding other element but also being able to further include other element if no contrary disclosure.

[33] FIG. 4 shows an SPR biosensor according to the first exemplary embodiment of the present invention. FIG. 4(a) is a perspective diagram illustrating the SPR biosensor according to the first exemplary embodiment of the present invention. FIG. 4(b) is a cross sectional view taken along line A-A' of FIG. 4(a).

[34] Referring to FIG. 4, the SPR biosensor 100 includes a sensor substrate 110 and a thin metal film 120. The sensor substrate 110 is an integration of a prism 111 and a flat type transparent dielectric substrate 112. The thin metal film 120 is formed on top of the surface of the prism 111 of the sensor substrate 110, and supports surface plasmons. [35] The sensor substrate 110 is formed of transparent optical polymer having a high refractive index and including polystylene (PS), polymethyl methacrylate (PMMA), polycarbonate (PC), and cyclic olefin copolymer (COC). The sensor substrate 110 can be manufactured in an injection molding method and the like.

[36] The thin metal film 120 is formed of metal such as aurum (Au), argentums (Ag), copper (Cu), and aluminum (Al) at a thickness of dozens of nanometers (nm), the real part of a dielectric function of which has a negative (-) value. Among the metals, argentums (Ag) which shows the sharpest SPR resonance peak and aurum (Au) which has excellent surface stability are used generally.

[37] The prism 111 makes light from any light source (not shown) go through and makes it totally reflected from the upper surface of the substrate just beneath the thin metal film 120, to couple light with the surface plasmons in metal 120. The prism 111 may have hemispherical, triangular, or trapezoidal from. In more detail, the sensor substrate 110 can have the same shape and size as a widely used slide glass format, for the convenience of use. For instance, the sensor substrate 110 is formed in a rectangular shape. That is, the prism 111 is formed at one side on a bottom surface of the rectangular sensor substrate 110 which has a predetermined thickness. The thin metal film 120 is formed on top of the sensor substrate 110 of which the position is normal to the prism 111.

[38] The SPR biosensor 100 can detect, by monitoring the changes in resonance angle, a biochemical reaction such as selective binding or unbinding between the biomolecules by proper chemical treatment of the metal 120. The SPR biosensor 100 can measure the strength of the specific binding and unbinding and the concentration of the biomolecules with a monochromatic light, which is incident on the thin metal film 120 through the prism 111, then reflected from the thin metal film 120, and then emitted through the prism 111.

[39] That is, an SPR biosensor system can be realized by connecting the SPR biosensor

100 with a light source, a light receiving unit, a polarizer, an goniometer, and a signal processor. The light source provides an incident light having a feature of a monochromatic or a polychromatic wavelength. The light receiving unit receives a reflected light from the prism 111. The polarizer transmits the incident light or the reflected light in a TM mode at a rear end of the light source or at a front end of the light receiving unit. The incidence angle controller controls an angle of incidence of the light that is incident on the prism 111 from the light source. The signal processor analyzes a signal from the light receiving unit and detects, by a change of an SPR angle, a change of an effective thickness or effective refractive index of a sample, on top of the thin metal film 120.

[40] [41] The incidence angle controller can be selectively added depending on a structure and principle of the biosensor system. For instance, the incidence angle controller can be realized as a driver for moving the light source in a mechanical method and controlling the angle of incidence or a lens for controlling the focus of the incident light.

[42] FIG. 5 shows an SPR biosensor according to the second exemplary embodiment of the present invention. FIG. 5 (a) is a perspective diagram illustrating the SPR biosensor according to the second exemplary embodiment of the present invention. FIG. 5(b) is a cross sectional view taken along line B-B' of FIG. 5 (a).

[43] Referring to FIG. 5, the SPR biosensor 200 includes a sensor substrate 110, a thin metal film 120, and one or more channels 130. The sensor substrate 110 is an integration of a prism 111 and a flat type transparent dielectric substrate 112. The thin metal film 120 is formed on top of the surface of the prism 111 of the sensor substrate 110, and supports surface plasmons. The one or more channels 130 are formed on top of the thin metal film 120 such that which are formed on the thin metal film across the SPR absorption dip line, at which a reflected light is minimized by plasmon resonant absorption of incident light by the thin metal film 120. The one or more channels 130 may have different effective refractive index or effective thickness change by different kind of coupling with biomolecules.

[44] That is, the SPR biosensor 200 is formed by further forming one or more channels

130 on the SPR biosensor 100 according to the first exemplary embodiment of the present invention. Other constituent elements than the channel 130 are realized identically with the first exemplary embodiment of the present invention.

[45] The one or more channels 130 are formed of dielectric materials in which part or all of each channel has different refractive index. By a change of refractive indices of the dielectric materials adsorbed on one or more channels 130, the change of absorption band by SPR of the thin metal film 120 occurs. This can be used to measure a change of an effective thickness and/or effective refractive index on the thin metal film 120.

[46] FIG. 6 is a schematic diagram illustrating the sensing principle using the SPR biosensor 200 according to the second exemplary embodiment of the present invention.

[47] In FIG. 6, light from any light source is extended to form a parallel, collimated two- dimensional light source 150 and the light is impinged on the prism 111 of the SPR biosensor 200 so that a central line of the prism 111 becomes a focal line. That is, the incident light is always normal to the surface of a prism 111.

[48] Thin metal film 120 is formed to have a thickness of about dozens of nanometers

(nm) (generally, 40 nm to 50 nm) on a top of the sensor substrate 110 opposite to the prism 111 including the focal line of the incident light to couple the surface plasmons of the thin metal film 120. Thin film 120 is made of metal (e.g., aurum (Au), argentums (Ag), copper (Cu), aluminum (Al) and the like) whose imaginary part of di- electric function (ε) in a wavelength range for observation has a negative (-) value. The dielectric function (ε) is defined by the refractive index (n) and the extinction ratio (k) as in Equation 1 below: [49]

[⁵°] ε = ε'+iε"= (n + ik)² = {n² - k²) + link ( 1)

[51] where,

[52] ε '

: real part of dielectric function, n²-k², and [53] ε"

: imaginary part of dielectric function, 2nk. [54]

[55] Meantime, a light totally reflected from the focal line in which all the incident light is converged is reflected again within an angle range of

#max in the same shape as the incident light. The reflected light can be integrated in a two- dimensional plane form using a cylindrical lens and a mirror. The collected reflected light appears as a two-dimensional plane image using a two-dimensional light receiving unit, a projection screen, or other elements and devices. The two-dimensional light receiving unit can be a Charge-Coupled Device (CCD) or a Complementary Metal Oxide Semiconductor (CMOS) imaging sensor, for example. The other elements and devices can detect brightness of each points of the two-dimensional plane.

[56] FIG. 7 is a graph showing the reflectivity as a function of the angle of incidence according to the first exemplary embodiment of the present invention.

[57] Referring to FIG. 7, in the case of the first exemplary embodiment of the present invention in which the channel 130 is not formed on the thin metal film 120, the minimum reflectivity of a reflected light obtained at a resonance absorption angle (

"sPR

) by SPR of the thin metal film 120 takes the shape of one resonance absorption band. Here, if brightness of each point can be absolutely distinguished, the resonance absorption band can be expressed using an absorption line in the end. Thus, each point on the absorption line is the minimum point (that is, a resonance dip) of reflectivity caused by SPR absorption. In principle, the shape of the cross section of the absorption band or absorption line should be equal to an SPR absorption curve 51 in a single channel shown in FIG. 7. [58] Alternatively, in the case of the second exemplary embodiment of the present invention in which one or more channels 130 are formed on the thin metal film 120 across the SPR absorption dip line and the dielectric having a small thickness is provided along the channel 130, a change of an SPR angle for each channel depending on the thickness of the dielectric on the thin metal film 120 occurs. The change of the SPR angle for each channel appears as a change of a position of the absorption line in a two-dimensional plane image of a reflected light that is detected from the SPR biosensor 200 according to the second exemplary embodiment of the present invention. [59] FIG. 8 is a graph illustrating the reflectivity versus incident angle of light in a SPR biosensor, according to the second exemplary embodiment of the present invention. Referring to FIG. 8, it is to be understood that the change of SPR angle (

AΘ_SPR

) takes place individually for each channel 130. Thus, as shown in FIG. 9, the SPR biosensor 200 has a plurality of channels 131 to 13n on the thin metal film 120. Biomolecules 311, 321, and 331 of a different kind, each are fixed to the channels 131 to 133, are specifically coupled with target molecules 312, 322, and 332 which exist in whole blood, in blood plasma, in serum, in urine, in saliva, and in other bio-samples that would be measured. If so, the change of surface concentration of each channel 131 to 133 occurs because of the coupling of the biomolecules. The surface concentration change in each channel 131 to 133 can be estimate from the change of effective thickness or effective refractive index of a sample, which is bound on the metal surface 120. Additionally, if well-known values of refractive indices of materials are used, a quantitation is also possible through an appropriate calibration. Also, the surface concentration change can be expressed by two-dimensional plane imaging of a reflected light emitted from the prism 111 of the SPR biosensor 200.

[60] The two-dimensional plane imaging of the reflected light using the SPR biosensor

200 is different from a Surface Plasmon Resonance Imaging (SPRI) method. In the SPRI method, a monochromatic light and a triangular prism are used and an angle of incidence of light is fixed to read a change of an effective thickness or effective refractive index ofdielectric, which is additively added on a metal surface and spatially distributed in a different way, using an intensity difference (e.g., a difference image) of a reflected light of each point (e.g., an image pixel).

[61] In the SPRI, the difference in brightness depends not only on thickness or refractive index changes, but also on other optical variables, e.g., on the wavelength or on incident angle.

[62] However, two-dimensional plane imaging of the reflected light using the SPR biosensor 200 is basically the same as an SPR angle interrogation method. In the SPR angle interrogation method, one basically measures the changes of SPR dip while fixing the wavelength of an incident light in one channel. Also, if one knows exactly the functional values of the refractive index of dielectric on metal film 130, even the absolute thickness change of the dielectric can be calculated by Fresnel equation.

[63] Thus, a SPR biosensor system can be constructed to include the SPR biosensor 200, the light source, the polarizer, and the two-dimensional imaging unit. The light source provides an incident light incident on the prism 111. The polarizer decomposes the light impinged on the prism 111 or the reflected light emitted from the prism 111 in a TM mode. The two-dimensional imaging unit processes, by two-dimensional plane imaging, the reflected light emitted through the prism 111. In the SPR biosensor system, the change of the effective thickness and effective refractive index of a sample which exists on top of the thin metal film 120 can appear as the change of absorption band appearing in the two-dimensional plane image.

[64]

[65] While the present invention has been shown and described in connection with the exemplary embodiments, it will be apparent to those skilled in the art that modifications and variations can be made without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

[1] A Surface Plasmon Resonance (SPR) biosensor comprising: a sensor substrate configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body; and a thin metal film formed in a position, which faces the prism, of an opposite surface to a surface on which the prism of the sensor substrate is formed and generating SPR by light impinged on the prism.

[2] The biosensor of claim 1, wherein the sensor substrate is formed of any one selected from the optical polymer group having a high refractive index and consisting of polystylene (PS), polymethyl methacrylate (PMMA), polycarbonate (PC), and cyclic olefin copolymer (COC).

[3] The biosensor of claim 1, wherein the sensor substrate is formed using an injection molding process.

[4] The biosensor of claim 1, wherein the thin metal film is formed of any one of aurum (Au), argentums (Ag), copper (Cu), and aluminum (Al).

[5] The biosensor of claim 1, wherein the prism has a hemispherical, triangular, or trapezoidal form..

[6] The biosensor of claim 1, further comprising: one or more channels which are formed on the thin metal film across the SPR absorption dip line, at which a reflected light is minimized by Plasmon resonant absorption of incident light, and all or part of which is formed of different dielectric material.

[7] The biosensor of claim 6, wherein on all or part of the one or more channels, each with different kind of biomolecules are immobilized to couple specifically with target molecules so that the surface concentration of complex molecules can be detected for each channel.

[8] A Surface Plasmon Resonance (SPR) biosensor system comprising: an SPR biosensor comprising: a sensor substrate configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body; and a thin metal film supporting surface plasmons and formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed, wherein the SPR biosensor makes light impinged on the prism into the upper surface of the substrate just beneath the thin metal film, and performs the total internal reflection above the critical angle; a light source for providing the incident light incident on the prism; a light receiving unit for receiving the reflected light, which is emitted through the prism, from the center of the prism and converting the reflected light into an electrical signal; a polarizer for decomposing the incident or the reflected light emitted from the prism, in a Transverse Magnetic (TM) mode; and a signal processor for analyzing the electrical signal from the light receiving unit and detecting a change of an effective thickness or effective refractive index of a sample, on top of the thin metal film by a change of an SPR angle.

[9] The system of claim 8, further comprising: an incident angle controller for controlling the angle of the incident light.

[10] The system of claim 9, wherein the incident angle controller is a goniometer or a rotation stage for moving the light source in a mechanical way and controlling the incident angle.

[11] The system of claim 9, wherein the incident angle controller is comprised of a lens controlling a focus of the incident angle.

[12] The system of claim 8, wherein the sensor substrate is formed of any one selected from the optical polymer group having a high refractive index and consisting of polystylene (PS), polymethyl methacrylate (PMMA), polycarbonate (PC), and cyclic olefin copolymer (COC).

[13] The system of claim 12, wherein the sensor substrate is formed using an injection molding process.

[14] The system of claim 8, wherein the thin metal film is formed of any one of aurum (Au), argentums (Ag), copper (Cu), and aluminum (Al).

[15] The system of claim 8, wherein the prism has a hemispherical, triangular, or trapezoidal form, makes light go through and makes it totally reflected from the upper surface of the substrate just beneath the thin metal film to perform the total internal reflection (TIR).

[16] The system of claim 8, wherein further comprising: one or more channels which are formed on the thin metal film across the SPR absorption dip line at which a reflected light is minimized by SPR of the thin metal film and all or part of which is formed of different dielectric material.

[17] The system of claim 16, wherein on all or part of the one or more channels, each with different kind of biomolecules are immobilized to couple specifically with target molecules so that the surface concentration of complex molecules can be detected for each channel.

[18] A Surface Plasmon Resonance (SPR) biosensor system comprising: an SPR biosensor comprising: a sensor substrate configured to have a prism and a flat type transparent dielectric substrate formed of the same material in a body; a thin metal film supporting surface plasmons and formed in a position, which faces the prism, on an opposite surface to a surface on which the prism of the sensor substrate is formed; and one or more channels which are formed on the thin metal film across the SPR absorption dip line, at which a reflected light is minimized by SPR of the thin metal film and all or part of which is formed of different dielectric material, wherein the SPR biosensor makes light impinged on the prism into the upper surface of the substrate just beneath the thin metal film, and performs the total internal reflection above the critical angle; a light source for providing the incident light on the prism; a polarizer for decomposing the incident or the reflected light emitted from the prism, in a Transverse Magnetic (TM) mode; and a two-dimensional imaging unit for imaging the reflected light emitted through the prism, as a two-dimensional plane image.

[19] The system of claim 18, wherein the incident light has a parallel two-dimensional light source form.

[20] The system of claim 19, further comprising: a signal processor for receiving a two-dimensional plane image signal and detecting an effective thickness or effective refractive index of a sample, on top of the thin metal film from a change of an absorption line.

[21] The system of claim 18, wherein the sensor substrate is formed of any one selected from the optical polymer group having a high refractive index and consisting of polystylene (PS), polymethyl methacrylate (PMMA), polycarbonate (PC), and cyclic olefin copolymer (COC).

[22] The system of claim 18, wherein the sensor substrate is formed using an injection molding process.

[23] The system of claim 18, wherein the thin metal film is formed of any one of aurum (Au), argentums (Ag), copper (Cu), and aluminum (Al).

[24] The system of claim 18, wherein the prism has a hemispherical, triangular, or trapezoidal form, makes light go through and makes it totally reflected from the upper surface of the substrate just beneath the thin metal film to perform the total internal reflection (TIR).

[25] The system of claim 18, wherein on all or part of the one or more channels, each with different kind of biomolecules are immobilized to couple specifically with target molecules so that the surface concentration of complex molecules can be detected for each channel.