KR102056971B1 - Apparatus and method for double prism solution immersed silicon biosensor - Google Patents

Abstract

The present invention relates to a double prism silicon-based liquid immersion microfluidic measuring apparatus and a measuring method, and more particularly formed on the support and the support and formed with a sample detection layer on which the first biological binding material is fixed to detect the first sample. A microchannel structure including at least one microchannel; A sample injection unit for injecting a buffer solution containing the first sample into the micro channel; A double prism unit disposed on the microfluidic structure and composed of an incident prism having a first refractive index and a reflective prism having a second refractive index; A polarization generating unit generating polarized light; And a polarization detector configured to detect a polarization change of the second reflected light, wherein the polarized light penetrates the incident prism to form incident light incident on the interface between the incident prism and the buffer solution, and part of the incident light is incident. The first reflected light is reflected at the prism-buffer solution interface to pass through the incident prism or the reflected prism, and another portion of the incident light passes through the buffer solution and enters the sample detecting layer at an incident angle satisfying the p-polarized wave antireflection condition. Forming a first transmitted light, the first transmitted light is reflected from the sample detection layer to form a second reflective light passing through the reflective prism-buffer interface and the reflective prism in contact with the buffer solution and the reflective prism, Dual prism silicon-based liquid, characterized in that the traveling direction of the first reflection light and the second reflection light is not parallel to each other It relates to a micro channel measuring device.

Description

Dual prism silicon-based immersion microfluidic measuring device and measuring method {APPARATUS AND METHOD FOR DOUBLE PRISM SOLUTION IMMERSED SILICON BIOSENSOR}

The present invention relates to a dual prism silicon-based immersion microfluidic measuring apparatus and a measuring method, and more particularly, using a double prism unit having a different refractive index, the first reflected light reflected from the sample detection layer and the prism-buffer solution The present invention relates to a dual prism silicon-based immersion microfluidic measuring device in which the traveling directions of the second reflected light reflected from the interface are not parallel to each other.

Reflectometry and ellipsometry are optical analysis techniques that determine the thickness or optical properties of a sample by measuring the change in reflectance or polarization state of the reflected light reflected from the surface of the sample and analyzing the measured values.

As measurement equipment using this, there is a reflectometer and an ellipsometer. They are used to evaluate various thin film thickness and physical properties of nano thin film manufacturing process in semiconductor industry. In addition, efforts are being made to extend the scope of application to the bio industry and apply it to interface analysis of bio materials such as proteins, DNA, viruses, and new drugs.

Conventional reflectometers are sufficient to evaluate the thickness and physical properties of nano-films with dimensions of several nanometers (nm) or more, but have low sensitivity in analyzing low molecular weight biomaterials that require sensitivity in the range of approximately 1 to 0.001 nanometers. There is a problem that the reliability is lowered. The ellipsometer has a measurement sensitivity of less than 0.01 nm compared to the reflectance measuring instrument. In particular, the measurement sensitivity is high under conditions where the refractive index is large, such as the thickness of an oxide film having a smaller refractive index than that of a semiconductor on a high refractive index semiconductor substrate.

However, in the case of an ellipsometer, a measurement method with improved sensitivity is required to analyze even low molecular weight biomaterials.

As a conventional technique for improving measurement sensitivity when analyzing a biomaterial, a surface plasmon resonance sensor (hereinafter referred to as an 'SPR sensor') in which a reflectance measuring method and a surface plasmon resonance (SPR) technology is mixed have.

Surface plasmon resonance (SPR) is a phenomenon in which electrons present on a metal surface are excited by light waves, causing collective vibration in the longitudinal direction of the surface, where light energy is absorbed. Say SPR sensor can measure the change of thickness and refractive index of nano thin film in contact with metal surface by using surface plasmon resonance which is sensitive to the polarization property of light. It is known that non-labeling can be measured in real time.

SPR sensor uses the principle that the structure of the glass is coated with a metal thin film of tens of nanometers on the material such as glass and the biomaterial can be bonded on it, and the resonance angle changes when the sample dissolved in the buffer solution is bonded to the sensor. The resonance angle is achieved by measuring the reflectance. When light enters the SPR sensor, the glass material becomes the incident medium and passes through the thin film layer to which the biomaterial is bonded. Finally, the buffer solution corresponds to the substrate.

In such a structure, the refractive index of the complete solution corresponding to the substrate material directly affects the movement of the resonance angle, similar to the change of the biological thin film layer due to the bonding of the sample to be measured. Therefore, in order to measure only pure bonding dynamics, the refractive index of the buffer solution should be measured and corrected independently.

In order to correct the refractive index change of the buffer solution and to prevent errors due to diffusion between the sample and the buffer solution, a sophisticated valve device, an air injection device, and a method of using one as a reference channel using two or more channels and correcting It is used. However, it is difficult to distinguish between the SPR angle change due to the refractive index change of the buffer solution and the SPR angle change due to the pure adsorption and dissociation characteristics, and can always act as a measurement error factor. As a result, the conventional SPR sensor has a fundamental difficulty in measuring the adsorption and dissociation characteristics of a material having a low molecular weight, such as a low molecule, due to the limitation of the above measurement method.

In addition, the conventional SPR sensor uses a metal thin film of a noble metal such as gold (Au), silver (Ag) for the surface plasmon resonance is expensive manufacturing of the sensor. In addition, the metal thin film has an uneven surface roughness depending on the manufacturing process, and the variation in refractive index is severe, and because of unstable optical properties, it is difficult to quantitatively measure biomaterials and to detect errors due to different sensitivity characteristics at different positions when compared with reference channels. There is a problem to include.

To improve the shortcomings of the SPR sensor, a biomaterial junction sensor layer is formed on a substrate material such as silicon and the ellipsometric measurement of the amplitude and phase of the light reflected from the substrate material through the buffer solution under the immersion microenvironment under the p-polarization antireflection conditions When measured by the method, the measured amplitude is not sensitive to the change in the refractive index of the buffer solution and a signal sensitive to the bonding dynamics of the biomaterial can be obtained. In the case of measuring the bonding characteristics of the biomaterial adsorbed to the substrate material under the immersion microfluidic environment, the buffer solution becomes the incident medium and the light passing through the biomaterial adsorption layer is reflected from the substrate material as opposed to the SPR measurement.

Under these measurement conditions, the elliptic measurement angle Ψ representing the measured amplitude is not sensitive to the change in the refractive index of the incident medium, which is a buffer solution, but only a change in the biofilm and substrate material. In the case of a stable refractive index substrate such as silicon, the measured ellipsometric angle Ψ can obtain a signal that is sensitive only to changes in the biofilm. In the case of using the prism incidence structure as shown in FIG. 1, the elliptic measurement angle Δ representing the image represents a signal sensitive only to the refractive index of the buffer solution, thereby simultaneously measuring the thickness of the biofilm and the refractive index of the buffer solution. However, when using a substrate parallel to a planar incidence structure such as a prism, the light reflected from the interface between the prism and the buffer solution should be removed and only the light reflected from the substrate should be used. In order to minimize the amount of sample used, the distance between the surface of the prism and the substrate material should be reduced. In this case, the two reflecting lights are located at a very close distance, which makes it difficult to separate and cause a measurement error. Therefore, in a planar incident structure such as a prism, a new structure measuring method is required to distinguish the light reflected from the interface between the prism and the buffer solution and the light reflected from the substrate material including the sensor.

1 is a cross-sectional view showing a biomaterial bonding characteristic measuring sensor according to the prior patent. As shown in FIG. 1, the biomaterial bonding characteristic sensor according to the prior patent includes a prism 100, a micro flow path structure 200, a polarization generator 300, and a polarization detector 400. At this time, the microfluidic structure 200 of the biomaterial bonding characteristic sensor according to the prior patent puts the adsorption layer 530 on the substrate 510 or the dielectric thin film 520, thereby forming an immersion microfluidic 210 environment. In this case, when the buffer 50 in which the sample 1 of the biomaterial is dissolved is injected into the micro channel 210, the biomaterial is adsorbed onto the ligand 2 formed on the surface of the adsorption layer 530. Thus, an adsorption layer having a predetermined thickness is formed.

The polarized incident light generated from the polarization generator 300 is incident on the interface between the buffer solution 50 and the substrate 510 at an angle causing the p-polarized wave antireflective condition to pass through the incident surface 110 of the prism. In this case, the reflected light reflected from the substrate 510 includes optical data regarding the refractive index of the adsorption layer and the buffer solution of the sample 1. That is, in the process of adsorbing and dissociating the sample 1 to the ligand 2, the molecular adsorption and dissociation kinetics such as the adsorption concentration, the thickness or refractive index of the adsorption layer, and the refractive index of the buffer solution are changed, As a result, the measured elliptic measurement angles are changed. The reflected light including the optical data is detected by the polarization detector 400. In this case, the polarization detector 400 may determine the molecular adsorption and dissociation dynamics of the sample 1 and the refractive index of the buffer solution by measuring the change according to the polarization component of the reflected light, that is, the ellipsometric angles.

2 shows an adsorption curve showing a process in which the sample 1 is adsorbed to the metal thin film 30, and a dissociation curve showing a dissociation process. The larger the association rate constant (ka), the faster the absorption of the biomaterial, and the smaller the dissociation rate constant (kd), the slower the dissociation.

That is, the dissociation constant (KD = kd / ka) at equilibrium can be obtained by measuring the adsorption rate constant and dissociation rate constant. For example, it is possible to determine whether a low-molecular drug candidate that can be used as a carcinogen can be used as a new drug by measuring the adsorption or desorption property of a protein including a carcinogenic factor.

Hereinafter, the features and limitations of the biomaterial analysis sensor according to the preceding characteristics will be described with reference to FIGS. 3 and 4. If to the incident light using a prism joining structure shown in Fig. 3 when substantially θ ₂ = the angle of inclination of 72.14 ° degree is incident on the living body thin film boundary surface is incident to the buffer solution in the prism by the change in refractive index (0.0002) in buffer solution An angle change of about -0.026 ° appears. The p-polarization antireflective condition is near θ ₂ = 72.14 °, but the current angle due to the change in the buffer refractive index is changed to 0.026 ° to 72.114 °, so as shown in FIG. Therefore, since the p-polarized light reflection angle is hardly changed, the values of Ψ and △ are measured at 72.114 °, which is a small angle of 0.026 °.

In FIG. 4, when the refractive indexes of the buffer solution 50 are different from each other, the solid line graph has a refractive index of 1.3330 and the dotted line graph corresponds to the refractive index of 1.3332 of the buffer solution 50. When the prism structure is used, the change of Ψ value is almost unchanged as shown in FIG. 4 due to the change of the refractive index of the buffer solution, while Δ shows a large change. That is, since the elliptic measurement constant Δ of the phase difference shows a sensitive change only in the refractive index change of the buffer solution and is hardly affected by the bonding property, only the change in the refractive index of the buffer solution can be measured with high sensitivity. The change in elliptic measurement constant △ shows a very large change as the thickness of the thin film material becomes very small, and when used in an applied study for analyzing the change in material properties or bonding properties by measuring the change in refractive index, It is a measuring method that can measure high sensitivity refractive index.

When a buffer solution having a different refractive index, such as a buffer solution continuously supplied and a solvent used in a sample, is supplied to a sensor through a microchannel, pure bonding dynamics and a change in the refractive index of the buffer solution can be simultaneously measured.

However, in FIG. 3, when the distance between the bottom of the prism and the substrate material is small, it is difficult to separate the light reflected from the interface between the prism and the buffer solution and the light reflected from the substrate material. Since the measurement is performed under the anti-reflective condition of p-polarized light, the intensity of light reflected from the substrate material is relatively weaker than the light reflected at the interface between the prism and the buffer solution, which may cause a measurement error.

Republic of Korea Patent Publication No. 10-1105328 Republic of Korea Patent Publication No. 10-1383652

Accordingly, the present invention has been made to solve the above problems, the problem to be solved by the present invention, by applying a double prism structure to the light reflected from the interface between the prism and the measurement medium and the light reflected from the substrate material By completely separating, it provides a dual prism silicon-based immersion microchannel measuring device capable of high sensitivity measurement.

Another object of the present invention is that in the conventional measuring method, the light reflected from the interface between the prism and the measuring medium is larger in energy than the light reflected from the substrate material and is difficult to separate, which may cause a measurement error. It is difficult to measure for other angles that change as the refractive index changes. The patent provides a dual prism silicon-based immersion microfluidic measuring device capable of efficiently separating the light reflected from the interface between the prism and the measurement medium.

In particular, in order to minimize the sample consumption, the channel height is reduced to the minimum, and the microchannel of the multi channel is provided, thereby providing various types of experimental conditions such as changing the concentration of the sample and varying the degree of adsorption of the self-assembled monolayer film. It is to provide a dual prism silicon-based immersion microchannel measuring device and a measuring method that can be.

Still another object of the present invention is to measure a high sensitivity of a biocompatible material in a non-labeled manner under an immersion microfluidic environment, thereby measuring a double prism silicon-based immersion microfluid that can be widely used in various industries such as bio, medical, food, and environment. It is to provide an apparatus and a measuring method.

However, the technical problems to be achieved in the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned are clearly to those skilled in the art from the following description. It can be understood.

As a means for achieving the above technical problem, the micro-channel structure including at least one micro-channel formed on the support and the sample detection layer formed on the support and fixed to the first biological binding material for detecting the first sample ; A sample injection unit for injecting a buffer solution containing the first sample into the micro channel; A double prism unit disposed on the microfluidic structure and composed of an incident prism having a first refractive index and a reflective prism having a second refractive index; A polarization generating unit generating polarized light; And a polarization detector configured to detect a polarization change of the second reflected light, wherein the polarized light penetrates the incident prism to form incident light incident on the interface between the incident prism and the buffer solution, and part of the incident light is incident. The first reflected light is reflected at the prism-buffer solution interface to pass through the incident prism or the reflected prism, and another portion of the incident light passes through the buffer solution and enters the sample detecting layer at an incident angle satisfying the p-polarized wave antireflection condition. Forming a first transmitted light, the first transmitted light is reflected from the sample detection layer to form a second reflective light passing through the reflective prism-buffer interface and the reflective prism in contact with the buffer solution and the reflective prism, Dual prism silicon-based liquid, characterized in that the traveling direction of the first reflection light and the second reflection light is not parallel to each other Provides a micro channel measuring device.

In addition, in one embodiment, a dual prism silicon-based immersion microfluidic measuring device characterized in that the first and second refractive index is different from each other is possible.

In addition, in one embodiment, it may be a dual prism silicon-based immersion microfluidic measuring device characterized in that the mirror reflecting coating on the contact surface of the incident prism and the reflective prism.

In addition, in one embodiment, the dual prism silicon-based immersion microchannel measuring device further comprises an optical device for adjusting the size of the incident light incident through the prism.

Further, in one embodiment, the optical device further comprises an imaging device, wherein the imaging device controls the shape of the beam spot of incident light formed on the prism-buffer interface. More preferred are base immersion microfluidic measuring devices.

Further, in one embodiment, the imaging device is more preferably a dual prism silicon-based immersion microflow meter, characterized in that any one of a lens, lens system, and reflector.

Further, in one embodiment, the sample detection layer includes a substrate, a dielectric thin film formed on top of the substrate, and an adsorption layer formed on top of the dielectric thin film, wherein the adsorption layer comprises a first biological binding material for detecting the first sample. It can be a double prism silicon-based immersion micro flow measurement device characterized in that it is fixed.

In addition, in one embodiment, the second reflected light is preferably a dual prism silicon-based immersion microfluidic measuring device further comprises a light reflected from the dielectric thin film.

In addition, in one embodiment, the dielectric thin film is more preferably a dual prism silicon-based immersion micro-flow measuring apparatus, characterized in that composed of any one of a transparent semiconductor oxide film and a glass film.

Further, in one embodiment, the dual prismatic silicon-based immersion microfluidic measuring device is characterized in that the dielectric thin film is 0 to 10 mm in thickness.

In addition, in one embodiment, the polarization detector, the double prism silicon-based liquid immersion further comprises a concentration calculation unit for calculating the thickness or concentration of the first sample adsorbed on the adsorption layer based on the polarization change of the second reflection light Micro flow measurement devices are preferred.

In addition, in one embodiment, the microchannel structure further comprises a second prism silicon-based liquid immersion, characterized in that the second microchannel formed with a second sample detection layer is fixed to the second biological binding material for detecting the second sample. It can be a microfluidic measuring device.

As a means for achieving the above technical problem, the sample injection unit is a buffer solution to the micro-channel structure comprising at least one micro-channel formed with a sample detection layer is fixed to the first biological binding material for detecting the first sample Injecting the first step; A second step of adsorbing the first sample contained in the buffer solution to the first antibody of the sample detection layer; A third step of causing the polarization generator to generate polarization; A fourth step of forming polarized light passing through the incident prism to be incident on the incident prism-buffer interface between the incident prism and the buffer solution; Part of the incident light is reflected at the incident prism-buffer solution interface to form a first reflected light that passes through the incident prism or the reflected prism, and another portion of the incident light passes through the buffer solution at an incident angle that satisfies the p-polarized wave antireflective condition. A fifth step of forming a first transmitted light incident on the sample detection layer; A sixth step of reflecting the first transmitted light from the sample detection layer to form a second reflective light passing through the reflective prism-buffer interface and the reflective prism in contact with the buffer solution and the reflective prism; A seventh step of detecting, by the polarization detector, a change in polarization of the second reflected light; And an eighth step of detecting the concentration of the sample adsorbed on the sample detection layer based on the polarization change of the second reflected light, wherein the first and second reflected light emitted to the outside of the double prism unit are mutually opposite. The present invention provides a dual prism silicon-based immersion microfluidic measuring device which is not parallel.

In addition, in one embodiment, a double prism silicon-based immersion microfluidic measuring method is characterized in that the refractive index of the incident prism and the refractive index of the reflective prism unit are different from each other.

In addition, in one embodiment, it may be a double prism silicon-based immersion micro-flow measurement method characterized in that the mirror prism coating on the contact surface of the incident prism and the reflective prism.

Further, in one embodiment, between the third step and the fourth step, step 3-1 for adjusting the size of the incident light incident through the prism through the optical device; further comprising a double prism silicon-based The immersion microchannel can be measured.

Further, in one embodiment, the optical apparatus further includes a reflecting apparatus including any one of a lens, a lens system, and a reflecting mirror, and step 3-1 includes an imaging device for incident light in which an imaging apparatus is formed on a prism-buffer interface. Controlling the shape of the beam spot (Beam Spot); is preferably a double prism silicon-based immersion micro-flow measurement method further comprising.

According to an embodiment of the present invention, by applying a double prism structure to completely separate the light reflected from the interface between the prism and the measurement medium and the light reflected from the substrate material, it is possible to measure high sensitivity.

In addition, according to an embodiment of the present invention, another object of the present invention, in the conventional measuring method is that the light reflected from the interface between the prism and the measuring medium is more energy than the light reflected from the substrate material, it is difficult to separate the measurement error When using an aperture to separate, can be measured for different angles that change with the change in refractive index.

In particular, in order to minimize the sample consumption, the channel height is reduced to the minimum, and the microchannel of the multi channel is provided, thereby providing various types of experimental conditions such as changing the concentration of the sample and varying the degree of adsorption of the self-assembled monolayer film. There are features that can be done.

In addition, according to one embodiment of the present invention, in order to minimize the consumption of the sample, in particular, by lowering the height of the flow path to a minimum, and having a multi-channel micro-channel, by changing the concentration of the sample or the degree of adsorption of the self-assembled monolayer membrane There is an advantage that can provide a variety of experimental conditions, such as different.

In addition, according to an embodiment of the present invention, it is possible to measure a high sensitivity of the bio-binding material in a non-labeled manner under the immersion micro-channel environment, it can be widely used in various industries such as bio, medical, food, environment.

However, effects obtained in the present invention are not limited to the above-mentioned effects, and other effects not mentioned above will be clearly understood by those skilled in the art from the following description. Could be.

The following drawings, which are attached in this specification, illustrate preferred embodiments of the present invention, and together with the detailed description of the present invention, serve to further understand the spirit of the present invention. It should not be construed as limited to.
1 is a cross-sectional view showing a biomaterial bonding characteristic measuring sensor according to the prior patent,
Figure 2 is a schematic diagram showing the adsorption concentration change in the process of the sample is adsorbed, dissociated to the metal thin film,
Figure 3 is a schematic diagram of a prism incident silicon-based immersion micro-flow measurement sensor for explaining the problems of the prior art,
4 is a graph measuring elliptic measurement constants Ψ and △ due to the change of the refractive index of the biomaterial adsorption and buffer solution using a biomaterial bonding characteristic measurement sensor according to the prior patent,
Figure 5a is a schematic diagram illustrating the configuration of a dual prism silicon-based immersion microchannel measuring apparatus according to an embodiment of the present invention,
Figure 5b is a schematic diagram illustrating the configuration and operation of the immersion microfluidic measuring device using a single prism,
5C is a schematic diagram illustrating a phenomenon in which the traveling directions of the first and second reflected light are not parallel by the dual prism unit acting on the microfluidic measuring device of FIG.
6A is a perspective view of a double prism silicon-based liquid immersion microfluidic structure according to an embodiment of the present invention;
6B is an exploded perspective view of a double prism silicon-based immersion microfluidic structure according to an embodiment of the present invention;
7 is a perspective view for explaining in detail the dual prism unit and the first structure according to an embodiment of the present invention,
8 is a cross-sectional view for explaining in detail the second structure according to an embodiment of the present invention;
9 is a flowchart illustrating a method of simultaneously measuring molecular bonding characteristics and a refractive index of a buffer solution according to the present invention;
FIG. 10 is a mirror reflecting coated double prism silicon-based immersion microchannel measuring apparatus according to an embodiment of the present invention. FIG.
Figure 11 is an asymmetric double prism silicon-based liquid immersion microchannel measuring apparatus according to an embodiment of the present invention.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present invention. However, since the description of the present invention is only an embodiment for structural or functional description, the scope of the present invention should not be construed as being limited by the embodiments described in the text. That is, the embodiments may be variously modified and may have various forms, and thus, the scope of the present invention should be understood to include equivalents for realizing the technical idea. In addition, the objects or effects presented in the present invention does not mean that a specific embodiment should include all or only such effects, the scope of the present invention should not be understood as being limited thereby.

The meaning of the terms described in the present invention will be understood as follows.

Terms such as "first" and "second" are intended to distinguish one component from another component, and the scope of rights should not be limited by these terms. For example, the first component may be named a second component, and similarly, the second component may also be named a first component. When a component is referred to as being "connected" to another component, it should be understood that there may be other components in between, although it may be directly connected to the other component. On the other hand, when a component is said to be "directly connected" to another component, it should be understood that there is no other component in between. On the other hand, other expressions describing the relationship between the components, such as "between" and "immediately between" or "neighboring to" and "directly neighboring to", should be interpreted as well.

Singular expressions should be understood to include plural expressions unless the context clearly indicates otherwise, and terms such as "include" or "have" refer to features, numbers, steps, operations, components, parts, or parts thereof described. It is to be understood that the combination is intended to be present and does not exclude in advance the possibility of the presence or addition of one or more other features or numbers, steps, operations, components, parts or combinations thereof.

All terms used herein have the same meaning as commonly understood by one of ordinary skill in the art unless otherwise defined. Generally, the terms defined in the dictionary used are to be interpreted as being consistent with the meanings in the context of the related art, and should not be interpreted as having ideal or excessively formal meanings unless clearly defined in the present invention.

First Example  Configuration

Hereinafter, the configuration of the preferred embodiment with reference to the accompanying drawings will be described in detail.

Figure 5a is a schematic diagram illustrating the configuration of a dual prism silicon-based immersion microchannel measuring apparatus according to an embodiment of the present invention.

Referring to FIG. 5A, the dual prism silicon-based liquid immersion microchannel measuring apparatus according to an exemplary embodiment of the present invention may include a dual prism unit 100, a microchannel structure 200, a polarization generator 300, and a polarization detector ( It can be recognized by including 400).

One embodiment of the present invention is to measure the adsorption and dissociation dynamics of biomolecules, including small molecules, using an ellipsometric method, a buffer (50) containing a sample (not shown) It has a structure that is injected into the micro-channel structure 200. In this case, the microchannel structure 200 may include a microchannel 210 having multiple channels.

The dual prism unit 100 may be configured by combining the incident prism 110 and the reflective prism 120. In addition, the incident prism 110 and the reflective prism 120 may be composed of a plurality of different prisms, respectively. The refractive index n1 of the incident prism 110 and the refractive index n2 of the reflective prism 120 may be configured differently.

On the other hand, in one embodiment of the present invention, the incident prism 110 and the reflective prism 120 may be mainly used optical glass, for example, may be BK7 or SF10, but is not limited thereto. In addition, the incident prism 110 and the reflective prism 120 may be composed of a plurality of prisms, respectively.

The microchannel structure 200 is disposed under the dual prism unit 100 and includes at least one microchannel 210 in which the sample detection layer 500 is formed. The sample detection layer 500 is formed on the support and the support, and the first biological binding material for detecting the first sample is immobilized.

Specifically, in one embodiment of the present invention, the microchannel structure 200 includes a plurality of microchannels 210 which are passages through which the buffer solution including the sample is introduced or discharged. At this time, the width of the micro channel 210 has a micro scale of about several mm or less than 1 mm. In addition, each of the plurality of microchannels 210 includes an inflow channel 210a, a microchannel channel 210c, and an outlet channel 210b. That is, the first microchannel 210 is formed by connecting the first inflow channel 210a, the first microchannel channel 210c, and the first discharge channel 210b.

Meanwhile, the sample injection unit 500 injects or discharges the buffer solution 50 including the first sample into the micro channel.

The polarization generator 300 may generate polarized light and may include a light source 310 and a polarizer 320. In addition, a collimating lens 330, a focusing lens 340, or a first compensator 350 may be provided.

The polarizer 320 and the first compensator 350 may be configured to be rotatable or may further include other polarization modulating means. The incident light 10 includes both the p-polarized wave and the polarization component of the s-polarized wave. In order to increase the signal-to-noise ratio (SNR), the polarizer 320 may be aligned at an angle close to the p-polarized wave, and preferably the p-polarized light included in the second reflected light received by the polarization detector described below. The ratio of wave and s-polarized wave can be similar.

In the present invention, the incident light 10 should be irradiated to the sample detecting layer 500 at an incident angle θ that satisfies the p-polarized wave antireflection condition. In the ellipsometric equation, the complex reflection coefficient ratio (ρ) can be expressed as the ratio of the reflection coefficient ratio (Rp) of the p-polarized wave to the reflection coefficient ratio (Rs) of the s-polarized wave, that is, ρ = Rp / Rs. The p-polarized light antireflection condition refers to a condition in which the reflection coefficient ratio Rp of the p-polarized wave has a value close to zero. The p-polarized light antireflection condition is similar to the surface plasmon resonance condition of the conventional SPR sensor, and is a condition in which the measurement sensitivity of the present invention is maximized.

The light source 310 is a semiconductor laser diode (LD) including various lamps, light emitting diodes (LEDs), solid-, liquid-, gas-lasers, and laser diodes that emit monochromatic or white light in the infrared, visible or ultraviolet wavelength range. ) May be used. In addition, the light source 310 may have a structure capable of varying the wavelength according to the structure of the optical system. On the other hand, in the vicinity of the above-described p-polarized wave antireflection conditions, the size of the optical signal of the reflected light may be relatively small, in which case the signal-to-noise ratio is increased by irradiating light with a high amount of light using a laser having coherence. Highly sensitive measurements can be enabled.

The collimating lens 330 receives light from the light source 310 to provide parallel light to the polarizer 320. In addition, the focusing lens 340 may converge parallel light passing through the polarizer 320 to increase the amount of incident light. In addition, the first compensator 350 serves to retard the polarization component of the incident light.

The polarization detector 400 receives the second reflected light 15-1 to be described later and detects the polarization change. The reflected light 15-1 reflected by the sample detection layer 530 receives the light and detects the change in the polarization state. The polarization detector 400 may include an analyzer 410, a photodetector 420, and an operation processor 430. In addition, the second compensator 440 and the spectrometer 450 may be provided.

The analyzer 410 corresponds to the polarizer 320 and may include a polarizer to polarize the reflected light again to control the degree of polarization of the reflected light or the direction of the polarization plane. The analyzer 410 may be configured to be rotatable according to the structure of the optical system, or may further include polarization modulating means capable of performing a function such as phase change and cancellation of the polarization component.

The photodetector 420 detects polarized reflected light to obtain optical data, and converts it into an electrical signal. At this time, the optical data includes information on the change of the polarization state in the reflected light. The photodetector 420 may be a CCD solid-state image pickup device, a photomultiplier tube (PMT), or a silicon photodiode.

The processor 430 obtains an electrical signal from the photodetector 420 to derive the measured value. The arithmetic processor 430 includes a predetermined analysis program using a reflectance measuring method and an elliptic measurement method. The arithmetic processor 430 extracts and analyzes optical data converted into an electrical signal, so that the adsorption concentration and the adsorption layer 160 of the sample are absorbed. The measured values such as the thickness, the adsorption constant, the dissociation constant, and the refractive index are derived. In this case, it is preferable that the operation processor 430 derives a measurement value by obtaining elliptic measurement constants Ψ and Δ related to the phase difference of the elliptic measurement method in order to improve measurement sensitivity.

The second compensator 440 plays a role of retarding and adjusting the polarization component of the reflected light. The second compensator 440 may be configured to be rotatable or may further include other polarization modulating means.

The spectrometer 450 is used when the light source 310 is white light. This is for spectroscopy of the reflected light and for separating the reflected light having a narrow wavelength range and sending the reflected light to the photodetector 420. In this case, the photodetector 420 may obtain optical data regarding the distribution of reflected light with a two-dimensional image sensor such as a CCD solid-state image sensor.

Hereinafter, the micro channel structure 200 according to an embodiment of the present invention will be described in more detail with reference to FIGS. 6 to 8.

FIG. 6A is a perspective view of a dual prism silicon-based immersion microchannel structure according to an embodiment of the present invention, and FIG. 6B is an exploded perspective view of the dual prism silicon-based immersion microchannel structure according to an embodiment of the present invention. In addition, FIG. 7 is a perspective view for explaining the dual prism unit and the first structure according to an embodiment of the present invention in more detail, and FIG. 8 is a cross-sectional view for explaining the second structure according to an embodiment of the present invention in detail. .

6A and 6B, the microchannel structure according to the exemplary embodiment of the present invention includes a first structure 200a and a second structure 200b.

In this case, the first structure 200a is formed under the double prism unit 100. In particular, in one embodiment of the present invention, the dual prism unit 100 and the first structure 200a may be manufactured in one piece, but are not limited thereto. In addition, the first structure 200a and the second structure 200b may be separated from each other, and the second structure 200b includes the microchannel channel layer 200c.

Meanwhile, the first structure 200 may be made of a transparent material such as glass or transparent synthetic resin material. In this case, as an example of the synthetic resin material, an acrylic resin such as polymethyl methacrylate (PMMA) may be used. And silicon-based materials such as silicon phosphate polymer (PDMS, polydimethylsiloxane) may also be used.

In more detail, as shown in FIG. 7, the first structure 200a includes a plurality of inflow paths 210a formed on one side of the first structure 200a and a plurality of discharge paths formed on the other side of the first structure. 210b. In addition, the inflow passage 210a is connected from the first inlet 212 formed at one side of the first structure to the second inlet 214 formed at the lower portion of the first structure, and the discharge passage 210b is formed of the first structure. From the first outlet 216 formed in the lower portion, it is connected to the second outlet 218 formed on the other side of the first structure.

Meanwhile, each of the plurality of inflow passages 210a and the discharge passage 210b is formed to be connected to the plurality of microchannel channels 210c of the microchannel channel layer 200c formed in the second structure 200b. Specifically, the second inlet 214 is formed to be in contact with one side of the microchannel channel 210c, thereby connecting the first channel 210a and the microchannel channel 210c.

In addition, the first outlet 216 is formed to contact the other side of the micro-channel 210c, the discharge passage 210b and the micro-channel 210c may be connected.

That is, the first inflow path 210a, the first microchannel channel 210c, and the first discharge path 210b are formed to be connected to each other. Accordingly, the buffer solution 50 including the sample injected through the first inflow path 210a may be discharged to the first discharge path 200b after passing through the first microchannel channel 210c.

In other words, the second structure 200b according to the embodiment of the present invention includes a microchannel channel layer 200c, and the microchannel channel layer 200c includes a plurality of microchannel channels 210c. The microchannel channel layer 210c may be formed of an acrylic resin such as polymethyl methacrylate (PMMA), but is not limited thereto.

In addition, the second structure 200b according to the embodiment of the present invention includes a sample detection layer 500 on the bottom of the groove formed by the plurality of microchannel channels 210c. In this case, the sample detection layer 500 specifically includes a substrate 510, a dielectric thin film 520 and an adsorption layer 530 formed on the substrate.

The substrate 510 has a complex refractive index of about 4.1385 + i0.04119 at 532 nm, and silicon (Si) that provides constant and stable physical properties at low cost may be used. However, the present invention is not limited thereto, and a semiconductor or dielectric material other than silicon may be used as the material of the substrate 510.

A layer of dielectric thin film 520 is formed on top of the substrate. The dielectric thin film 520 may be a thin film material of a transparent material including a semiconductor oxide film or a glass film. At this time, the thickness of the dielectric thin film is preferably 0 ~ 1000nm.

On the other hand, the most easily obtained dielectric thin film 520 is a silicon oxide film (SiO 2) grown to several nanometers by naturally oxidizing silicon. Since the refractive index of the silicon oxide film is about 1.461 at 532 nm, the difference in refractive index with the substrate 510 made of silicon is large, which helps to increase the measurement sensitivity of the present invention.

In addition, a glass film made of optical glass may be used for the dielectric thin film 520. The dielectric thin film 520 formed of silicon, silicon oxide, or glass film can provide a stable optical characteristic and can provide stable optical characteristics and lower manufacturing cost compared to a metal thin film such as gold or silver. .

Adsorption layer 530 according to an embodiment of the present invention may be composed of at least one of a self-assembled thin film and a bio thin film. In addition, the adsorption layer 530 may be immobilized bio-binding material capable of detecting a specific sample.

In this case, the adsorption layer 530 serves to absorb and dissociate a sample (not shown) of the low molecular weight biocombination material and reflect incident light.

In other words, the sample contained in the buffer solution introduced through the first inflow path 210a may be adsorbed to the adsorption layer 530 and dissociated from the adsorption layer 530.

On the other hand, although not shown, one embodiment of the present invention may further include a sample injection unit. In this case, the sample injection unit (not shown) injects or discharges the buffer solution or the buffer solution containing the sample in the micro-channel 210.

First Example  action

Hereinafter, with reference to the accompanying drawings will be described in detail the operation of the preferred embodiment.

FIG. 5B is a schematic view illustrating the configuration and operation of the liquid immersion microfluidic measuring device using a single prism, and FIG. 5C illustrates the progress of the first and second reflected light by a dual prism unit acting on the microfluidic measuring device of FIG. 5A. It is a schematic diagram explaining the phenomenon that a direction is not parallel.

First, referring to FIG. 5B, a single prism structure composed of an incident prism 110 having a refractive index n ₁ is used. The incident light 10 penetrates the incident prism 110 and is incident on the incident prism-buffer interface 111. At the incident prism-buffer solution interface 111, the incident light 10 is separated into a first reflected light 14 and a first transmitted light 12. The second transmitted light 12 is reflected by the sample detection layer 500 and again passes through the incident prism-buffer interface 111 and the incident prism 110 to form the second reflected light 15. At this time, the angle between the first reflected light 14 and the second reflected light 15 and the vertical line, θ _t1 and θ _t1 are the same.

The first reflected light 14 and the second reflected light 15 are split from the same incident light 10. The first reflected light 14 and second reflected light 15, where each of the reflection angle of refraction caused by the refractive index n ₂ of a refractive index difference n ₁ with the buffer solution 50 is different in the incident prism 110 is the same. Therefore, even if the reflected positions are different from each other, the angles when emitted to the outside of the prism through the incident prism 110 are the same.

As described above, the photodetector 400 receives the second reflected light and analyzes its polarization state, thereby extracting property information related to the thickness or optical properties of the sample. However, as shown in FIG. 5B, a single prism structure When θ _t1 and θ _t1 are the same, it is difficult to separate the first reflected light 14 and the second reflected light 15. That is, in order to receive only the second reflected light 15 to the photodetector 400, there is a problem in that a separate optical system is disposed in front of the photodetector 400 or the beam spot size of the incident light 10 must be minimized. have.

Referring to FIG. 5C, by applying the dual prism unit 100, it may be confirmed that the traveling directions of the first and second reflective lights 14-1 and 15-1 are different from each other.

In the related art, it is difficult to separate the light 14 reflected from the interface between the prism 100 and the medium and the light 12 refracted and incident on the adsorption layer 530. Thus, the prism 100 and The light 14 reflected at the interface of the medium is refracted and detected by the polarization detector 400 together with the light 12 incident on the adsorption layer 530. Therefore, a measurement error is caused by the light 14 reflected from the boundary between the prism 100 and the medium having a relatively high energy, and the measurement sensitivity is deteriorated.

On the other hand, as shown in Figure 5c, the dual prism silicon-based immersion microfluidic measuring device according to an embodiment of the present invention, the incident light 10 is the first reflected light 14 at the incident prism-buffer solution interface 111 And the first transmitted light 12 are the same as those of the single prism of FIG. 5B. However, in the process of transmitting the first reflected light 14 and the first transmitted light 12 through the reflective prism 120, a difference occurs in each refractive angle. The refractive index of the reflecting prism (120), n ₃ is the refractive index n can be between ₁ and n are made different, and preferably _1> n _3> n ₂ of the incident prism 110 is formed. When n ₁ > n ₃ > n ₂ is established, Δθ _t1 and Δθ _t2 become large when n ₃ approaches n ₂ .

Since the first reflection light 14-1 and the second reflection light 15-1 are not parallel and are opened by an angle of Δθ _t1 + Δθ _t2 , the second reflection light 15-1 is separated and received by the photodetector 400. It is easy to do That is, when the photodetector 400 is positioned away from the double prism unit 100 by a specific distance, the separation distance between the second reflected light 15-1 and the first reflected light 14-1 becomes long. Therefore, the amount of light of the first reflected light 14-1 focused on the light detector 400 may be minimized. In addition, if the optical axis of the light detector 400 is aligned with the optical path of the second reflected light 15-1, the influence of the small amount of first reflected light 14-1 that is unnecessarily focused on the light detector 400 may be minimized. Can be. That is, the propagation loss of the first reflected light 14-1 is greater than the propagation loss of the second reflected light 15-1 while traveling inside the photodetector 400. Therefore, the amount of light of the first reflected light 14-1 reaching the photodetector 420 may be limited to a relatively small amount compared to the amount of light of the second reflected light 15-1.

In conclusion, the dual prism silicon-based immersion microfluidic measuring apparatus and measuring method according to an embodiment of the present invention includes a second reflected light 15-1 and a first reflected light 14-1 including characteristic information of a sample. By facilitating separation of the samples, not only the measurement error but also the high sensitivity analysis of the sample can be facilitated.

In addition, when the relationship of n ₃ > n ₁ is established in FIG. 5C, the first reflected light 14-1 and the second reflected light 15-1 cross each other and proceed. That is, since Δθ _t1 and Δθ _t2 have negative values, when the distance between the photodetector 400 and the reflective prism 120 is sufficiently secured, the second reflected light is similar to the case of n ₁ > n ₃ > n ₂ . Separation detection of (15-1) is easy.

Meanwhile, θ _i3 in FIG. 5C satisfies the p-polarized wave antireflection condition.

9 is a flowchart illustrating a method of simultaneously measuring molecular bonding characteristics and a refractive index of a buffer solution according to the present invention. As shown in Figure 9, the measuring method of the present invention is subjected to the first step (S100) to the eighth step (S500).

In the first step S100, as shown in FIG. 5, the sample injection unit dissolves a sample (not shown) of a biobinding material including a low molecule in the buffer solution 50 and injects it into the microchannel 210 of the microchannel structure 100. Done. In this case, the sample injector may inject the buffer solution 50 including the samples having different concentrations in each of the microchannels 210 of the multichannel.

In addition, the buffer solution 50 may be injected with a time difference for each microchannel 210. In addition, the buffer solution 50 may be injected only to some microchannels 210, and the other microchannels 210 may not be used.

In the second step S200, a sample (not shown) of a biocombination material is adsorbed onto the substrate 510 or the dielectric thin film 520 to form the adsorption layer 530.

Alternatively, a sample may be adsorbed onto a plurality of different self-assembled monolayer films or the same self-assembled monolayer film formed in the single microfluidic channel 210c of FIG. 8 to form adsorption layers having different bonding characteristics. It may be.

In a third step S300, the polarization generator generates polarization. The polarizer 320 polarizes predetermined light emitted from the light source 310 to generate polarized light.

In the fourth step S400, polarized light is transmitted through the incident prism 120 to form incident light incident on the incident prism-buffer interface 111 in contact with the incident prism and the buffer solution 50.

In the fifth step S500, a part of the incident light 10 is reflected from the incident prism-buffer interface 111 to form the first reflected light 14-1 passing through the incident prism 110 or the reflective prism 120. In addition, another part of the incident light 10 penetrates the buffer solution 50 to form the first transmitted light 12 incident on the sample detecting layer 500 at an incident angle satisfying the p-polarized wave antireflection condition.

In a sixth step 600, the first transmitted light 12 is reflected from the sample detection layer 500, such that the reflective prism-buffer interface 121 and the reflective prism in contact with the buffer 50 and the reflective prism 120 are provided. The second reflected light 15-1 penetrating 120 is formed.

In a seventh step 700, the polarization detector 400 detects a polarization change of the second reflected light.

The eighth step 800 detects the concentration of the first sample adsorbed on the sample detection layer based on the polarization change of the second reflected light.

Specifically, the analyzer 410 first receives the elliptically polarized reflected light from the adsorption layer 530 to pass only the light according to the polarization characteristics.

Next, the photodetector 420 obtains predetermined optical data by detecting a change in the polarization component of the reflected light, converts it into an electrical signal, and transmits the converted optical signal to the operation processor 430.

Next, an arithmetic processor 430 having a program using a reflectance measuring method or an ellipsometric method extracts and analyzes optical data converted into an electrical signal, and the adsorption concentration of the sample, the adsorption and dissociation constant, the refractive index, the refractive index of the buffer solution, The same measurement is derived.

At this time, in the present invention, the operation processor 430 obtains the elliptic measurement constant Δ of the phase difference of the elliptic measurement method, measures the refractive index measurement value of the buffer solution, and measures the elliptic measurement constant? The reason is that the elliptic measurement constant Δ of the phase difference in the p-polarized light antireflection condition is sensitive to the change in the refractive index of the buffer solution and is almost unaffected by the bonding properties. Therefore, only the change in the refractive index of the buffer solution can be measured. This is because the elliptic measurement constant Ψ with respect to the amplitude ratio changes mainly with high sensitivity to the bonding properties of the material.

Therefore, the bonding characteristics of the sample introduced into the buffer solution are measured as Ψ and the refractive index change of the buffer solution including the refractive index that changes when dissolved in the buffer solution or the solvent such as DMSO used to dissolve the sample is measured at the same time as Δ Only the bonding characteristics are obtained.

10 is an apparatus for measuring a mirror-reflective dual prism silicon-based liquid immersion microchannel according to an embodiment of the present invention, and FIG. 11 is an asymmetric double prism silicon-based immersion microchannel measuring device according to an embodiment of the present invention.

FIG. 10 is a structure in which a mirror reflective coating is applied to an interface between the incident prism 110 and the reflective prism 120. The first reflected light 14 is reflected by the incident prism 110, thereby traveling in a direction opposite to the second reflected light 15-1.

In addition, referring to FIG. 11, by applying an asymmetric double prism structure, the first reflected light 14-1 may be totally reflected on the surface of the reflective prism 120. That is, the angle θ _i2 is n ₁ , If the incident angle θ _i2 is controlled to be larger than the total reflection angle calculated based on n ₃ , the difference in the travel direction between the first and second reflected light 14-1 and 15-1 may be increased as shown in FIG. 10.

The detailed description of the preferred embodiments of the invention disclosed as described above is provided to enable those skilled in the art to implement and practice the invention. Although the above has been described with reference to preferred embodiments of the present invention, those skilled in the art will understand that various modifications and changes can be made without departing from the scope of the present invention. For example, those skilled in the art can use each of the components described in the above-described embodiments in combination with each other. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

The invention can be embodied in other specific forms without departing from the spirit and essential features of the invention. Accordingly, the above detailed description should not be construed as limiting in all aspects and should be considered as illustrative. The scope of the present invention should be determined by reasonable interpretation of the appended claims, and all changes within the equivalent scope of the present invention are included in the scope of the present invention. The present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein. In addition, the claims may be incorporated into claims that do not have an explicit citation relationship in the claims, or may be included as new claims by amendment after filing.

100: double prism unit
200: microfluidic structure
300: polarization generating unit
400: polarization detection unit

Claims (17)

A microchannel structure including a support and at least one microchannel formed on the support and having a sample detection layer on which a first biological binding material for detecting a first sample is immobilized;
A sample injection unit for injecting a buffer solution containing the first sample into the micro channel;
A double prism unit disposed on the microchannel structure, the double prism unit including an incident prism having a first refractive index and a reflective prism having a second refractive index;
A polarization generating unit generating polarized light; And
Including; a polarization detector for detecting a polarization change of the second reflected light
The polarized light is transmitted through the incident prism to form incident light incident on the interface between the incident prism and the buffer solution where the incident prism and the buffer solution come into contact with each other.
A portion of the incident light is reflected at the incident prism-buffer solution interface to form a first reflected light that passes through the incident prism or the reflected prism, and another portion of the incident light passes through the buffer solution to prevent p-polarized wave reflection. Forming a first transmitted light incident on the sample detecting layer at an incident angle satisfying the condition;
The first transmitted light is reflected by the sample detection layer to form a reflective prism-buffer boundary interface between the buffer solution and the reflective prism and a second reflected light passing through the reflective prism,
The traveling direction of the first reflected light and the second reflected light emitted to the outside of the double prism unit is not parallel to each other,
The sample detection layer
Board,
A dielectric thin film formed on the substrate, and
An adsorption layer formed on the dielectric thin film,
The first biological binding material for detecting the first sample is immobilized in the adsorption layer,
And a dual prism silicon-based liquid immersion microfluidic measuring device parallel to the incident prism-buffer solution interface and the reflective prism-buffer solution interface. The method of claim 1,
The device of claim 1, wherein the first and second refractive indices are different from each other. The method of claim 1,
The dual prism silicon-based immersion micro-flow measuring apparatus, characterized in that the mirror prism coating on the contact surface of the incident prism and the reflective prism. The method of claim 1,
The dual prism silicon-based immersion microchannel measuring apparatus further comprises an optical device for adjusting the size of the incident light incident through the prism. The method of claim 4, wherein
The optical device further comprises an imaging device,
And the imaging device controls a shape of a beam spot of the incident light formed on the incident prism-buffer solution interface. The method of claim 5,
The imaging device
Dual prismatic silicon-based immersion micro-flow measuring apparatus, characterized in that any one of a lens, a lens system, and a reflector. delete The method of claim 1,
The second reflected light further comprises a light reflected from the dielectric thin film dual prism silicon-based immersion micro-flow measuring apparatus. The method of claim 1,
The dielectric thin film is a dual prismatic silicon-based immersion micro-flow measuring apparatus, characterized in that composed of any one of a transparent semiconductor oxide film and a glass film. The method of claim 1,
The dielectric thin film
Dual prismatic silicon-based immersion microfluidic measuring device, characterized in that the thickness is greater than 0 ~ 10 mm. The method of claim 1,
The polarization detection unit,
And a concentration calculating unit calculating a thickness or a concentration of the first sample adsorbed on the adsorption layer based on the polarization change of the second reflected light. The method of claim 1,
The micro channel structure is
The dual prism silicon-based immersion microchannel measuring apparatus further comprises a second microchannel in which a second sample detection layer is immobilized with a second biological binding material for detecting the second sample. A first step of injecting a buffer solution into a microchannel structure including at least one microchannel on which a sample detection layer is immobilized with a first biological binding material for detecting a first sample;
A second step of adsorbing the first sample contained in the buffer solution to the first antibody of the sample detection layer;
A third step of causing the polarization generator to generate polarization;
A fourth step in which the polarized light is transmitted through the incident prism to form incident light incident on an interface between an incident prism and a buffer solution in contact with the incident prism and the buffer solution;
A portion of the incident light is reflected at the incident prism-buffer solution interface to form a first reflected light that passes through the incident prism or the reflected prism, and another portion of the incident light passes through the buffer solution to prevent p-polarized wave reflection. A fifth step of forming first transmitted light incident on the sample detecting layer at an incident angle satisfying a condition;
A sixth step of reflecting the first transmitted light from the sample detection layer to form a reflective prism-buffer boundary interface between the buffer solution and the reflective prism and a second reflected light passing through the reflective prism;
A seventh step of detecting a polarization change of the second reflected light by the polarization detector; And
And an eighth step of detecting a concentration of a sample adsorbed on the sample detection layer based on the polarization change of the second reflected light.
The advancing direction of the first reflected light and the second reflected light emitted to the outside of the double prism unit is not parallel to each other,
The sample detection layer
Board,
A dielectric thin film formed on the substrate, and
An adsorption layer formed on the dielectric thin film,
The first biological binding material for detecting the first sample is immobilized in the adsorption layer,
And a dual prism silicon-based immersion microfluidic measurement method, wherein the incident prism-buffer solution interface and the reflective prism-buffer solution interface are horizontally parallel to each other. The method of claim 13,
And a refractive index of the incident prism and a refractive index of the reflective prism are different from each other. The method of claim 13,
The double prism silicon-based liquid immersion micro-flow measurement method characterized in that the mirror prism coating on the contact surface of the incident prism and the reflective prism. The method of claim 13,
Between the third step and the fourth step,
And a third step of adjusting the size of the incident light incident through the prism through an optical device including an imaging device. 2. The method of claim 16,
The optical device
Further comprising a reflector comprising any one of a lens, a lens system, and a reflector,
Step 3-1
And a step of controlling, by the imaging device, a shape of a beam spot of the incident light formed on the incident prism-buffer solution interface.

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