JP2003065947A - Apparatus and method for measuring refractive index as well as qualitative and quantitative analyzer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To realize an apparatus and method for the measuring a refractive index which are not subjected to the influence of a temperature change, which do not require a complicated preparation process, which can measure the refractive index in real time, with high accuracy and with high sensitivity and which are simple and low-cost. SOLUTION: An AC voltage is applied across an electrode 12 and an electrode 13 as a pair, a dielectric migration force is made to act on molecules or particles contained in a solution 10 to be analyzed, and the molecules or the particles are collected on a first metal membrane 7 which generates surface plasmon resonance. Since their collection easiness to an electric-field concentration part is different according to the molecules or the particles, the refractive index can be measured with high accuracy and with high sensitivity when the frequency and the voltage value of the AC voltage are set to values optimum for the molecules or the particles.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a refractive index measuring device,
The present invention relates to a refractive index measuring method and a qualitative quantitative analyzer.

[0002]

2. Description of the Related Art Conventionally, as a method for measuring the refractive index of a solution to be analyzed in the vicinity of the surface of a metal thin film, there is a method utilizing a surface plasmon resonance phenomenon. This is because when the wave number of the surface plasmon induced by the incident light applied to the back surface of the metal thin film matches the wave number of the evanescent wave derived from the excitation light,
The energy of the incident light is used to excite surface plasmons,
The fact that the amount of reflected light is reduced is used. As the method, the change in the refractive index of the solution to be analyzed in the vicinity of the metal surface is measured by changing the incident angle of the monochromatic incident light and the reflected light is measured. There is a method in which the reflected light is measured by making the light incident at, and the change is obtained from the change in the wavelength at which the maximum absorption occurs. Further, by using the above-described refractive index measuring method, it is possible to configure a sensor that detects molecules and fine particles in the solution to be analyzed.

For example, Japanese Patent Laid-Open No. 6-58873 discloses an optical sensor which uses surface plasmon resonance to quantify a specific molecule in a sample solution which shows a specific reaction with a molecular recognition functional substance. Has been done. This is because the molecular recognition functional film is provided on the surface of the metal thin film facing the solution flow path through which the sample to be analyzed is provided, and the refractive index change of the specific molecule adsorbed on the molecular recognition functional film is obtained by using the surface plasmon resonance phenomenon. It is an invention relating to a sensor for obtaining a molecular concentration.

Further, it is known that the surface plasmon resonance phenomenon has temperature dependence, and if temperature changes during measurement, accurate measurement becomes difficult. On the other hand, JP-A-9-25
In Japanese Patent No. 7698, two surface plasmon resonance measurement channels are used, and one channel is used for measuring a target substance and the other channel is used as a reference standard, thereby compensating for fluctuations in the surface plasmon resonance phenomenon due to temperature changes. It is configured to do.

Further, conventionally, as a method for analyzing a solution to be analyzed containing various kinds of molecules and fine particles, a gel chromatography method in which an analysis is made by utilizing a difference in adsorption by gel, and a high frequency non-uniform electric field are generated. There is an electrostatic chromatography method in which analysis is performed by utilizing the difference in dielectrophoretic force acting on molecules and particles.

[0006] For example, in Japanese Patent No. 3097932, molecules and particles to be sampled are added to a carrier flowing at a constant speed from an inlet, and dielectrophoretic force is exerted on these to cause a difference in the time required to reach the outlet. An electrostatic chromatography device for the analysis of particles and particles is described.
This is an invention relating to a chromatography device that analyzes a molecule or particle by utilizing the fact that the dielectrophoretic force acting on the molecule or particle varies in size depending on the electric dipole moment peculiar to the molecule or particle.

[0007]

The above-mentioned conventional techniques are all aimed at establishing a method and apparatus for qualitatively and quantitatively analyzing molecules and particles. However, the above conventional technique has the following problems.

In Japanese Patent Laid-Open No. 6-58873, a molecular recognition functional film is provided on the surface of a metal thin film in which a surface plasmon resonance phenomenon occurs, so that a specific molecule that shows a specific reaction with the molecular recognition functional film is reliably captured and the Conducting sensitive qualitative and quantitative analysis. However, the complicated pretreatment process of providing the molecular recognition functional film on the metal thin film is costly and time consuming. Further, in biochemical analysis and immunological analysis targeting serum and plasma, serum and plasma contain molecules and particles such as proteins and cells of various types, and only specific molecules are analyzed above. It is very difficult to deal with with technology.

Further, in Japanese Patent No. 3097932,
We apply various dielectrophoretic forces to the molecules and particles contained in a carrier flowing at a constant velocity, and analyze various molecules and particles based on the difference in the time required to move a certain distance. However, since the analysis method is based on the difference in required time, the analysis will take some time.

Further, both techniques have a problem that the solution to be analyzed is caused to flow at a constant rate and a separate carrier solution is required.

Further, Japanese Unexamined Patent Publication No. 9-257698 describes an invention using two surface plasmon resonance channels in order to compensate for fluctuations in the surface plasmon resonance phenomenon due to temperature changes. However, providing two channels complicates the device and adds cost.

In view of the above, the object of the present invention is not to use a complicated process such as adding an analyte solution to a carrier and flowing it, or preparing a molecular recognition functional film that reacts only with specific molecules, and By providing a simple and inexpensive refractive index measuring device and measuring method capable of removing the fluctuation component of the measurement result due to change, capable of high-accuracy, high-sensitivity and high-speed measurement, and by providing a qualitative quantitative analyzer. is there.

[0013]

In order to solve this problem, the present invention provides a light source, a first metal thin film that causes surface plasmon resonance with light emitted from the light source, and a first metal thin film. A photodetector for receiving light, an electrode pair composed of a second metal thin film and a third metal thin film for dielectrophoresing the solution to be analyzed, a voltage source for generating an alternating electric field between these electrodes, and these Refraction of the solution to be analyzed, which has a transparent base material that supports the metal thin film, and a control calculation unit that controls the light source, photodetector, voltage source, etc. and calculates the refractive index from the measurement results obtained by the photodetector. This is a rate measuring device.By dielectrophoresing the solution to be analyzed, it becomes possible to position the desired molecules and particles on the metal thin film that causes surface plasmon resonance, without using a complicated pretreatment process. Do not run the analyte solution , It is possible to provide a convenient and highly sensitive refractive index measurement apparatus.

Here, the metal thin film which causes surface plasmon resonance may also have a function of an electrode for dielectrophoresing the solution to be analyzed.

Further, by measuring the temperature of the solution to be analyzed and / or the metal thin film and correcting the refractive index measured using the temperature, it is possible to remove the fluctuation component of the measurement result due to the temperature change, and it is possible to achieve high accuracy. A refractive index measuring device can be provided.

The temperature measuring means for the solution to be analyzed and / or the metal thin film may be obtained from the change in the electric resistance value of the metal thin film forming the electrode, or the temperature change of the solution to be analyzed and / or the metal thin film may be obtained. It may be detected directly by using a thermocouple, etc., or a metal wire made of a metal different from the metal thin film that causes surface plasmon resonance is connected to the metal thin film to change the temperature of the metal thin film due to the electromotive force generated at the interface between different metals. May be asked.

Further, when dielectrophoresis is generated, by controlling the frequency and / or voltage value and / or voltage application time of the alternating electric field applied, the types of particles and molecules trapped are controlled, and From the measurement result of the refractive index in the case, it becomes possible to obtain the refractive index due to each of the particles and molecules contained in the solution to be analyzed,
It is possible to provide a refractive index measurement method capable of measuring the refractive index of a plurality of types of particles or molecules not limited to a specific molecule at high speed.

Further, a comparison between the refractive index before the dielectrophoresis is generated and the refractive index immediately after the dielectrophoresis is generated, and /
Alternatively, by comparing the refractive index before the dielectrophoresis is stopped with the refractive index immediately after the dielectrophoresis is stopped, it is possible to use the temperature correction means for measuring the refractive index without being affected by the temperature change. Therefore, it is possible to provide a simple, inexpensive, and highly accurate refractive index measuring method without introducing a complicated device configuration.

Then, the refractive index obtained by using the above apparatus and method, the frequency of the AC voltage applied between the electrodes when the dielectrophoresis is generated, the voltage value of this AC voltage, and the application time of this AC voltage. From the above, it is possible to provide a qualitative quantitative analysis device for identifying the types of molecules and particles in the solution to be analyzed and / or measuring the number of molecules and particles in the solution to be analyzed.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below with reference to the drawings.

(Embodiment 1) FIG. 1 is a schematic diagram showing the measurement of the refractive index of a solution to be analyzed. In FIG. 1, incident light 5 emitted from a light source 1 passes through a first lens 2, a second lens 3, a prism 4 and a glass 6, and a first metal thin film 7
Is irradiated. The light source is a laser diode that oscillates a single wavelength, and the first lens 2 and the second lens 3 are adjusted so that the focal point of the incident light 5 is formed by the first metal thin film 7. That is, the incident light 5 has a lens size, a focal length,
It has an incident angle range defined by the refractive index of the prism.

The first metal thin film 7 is preferably an Au thin film, but other metals such as Ag, Cu, Al and Pt may be used as long as they cause the surface plasmon resonance phenomenon. Absent.

Further, the surface of the first metal thin film 7 is coated with a non-metallic substance having a thickness of 1000 nm or less in order to prevent the molecules and particles contained in the solution to be analyzed 10 from being adsorbed by the metal. Is preferred. 1000 thickness
When the thickness exceeds nm, the evanescent wave does not substantially propagate. In some cases, adsorption may not occur depending on the combination of the first metal thin film 7 and the molecules or particles in the solution to be analyzed 10, and in that case, the coating may be omitted.

Reflected light 8 reflected by the first metal thin film 7
Is again transmitted through the glass 6 and the prism 4 and is irradiated to the photodetector 9, where the light amount is detected for each incident angle. The photodetector 9 is preferably composed of a CCD or array sensor. The prism 4 and the glass 6 are in close contact with each other with a matching oil (not shown).

The glass 6 forms a part of a container 11 for holding the solution to be analyzed 10. In FIG. 1, the entire bottom surface of the container 11 is made of glass, but the entire container 11 may be made of glass, or only the portion that transmits the incident light 5 and the reflected light 8 may be made of glass.

Using the optical measurement system described above, reflected light 8
The angle at which the amount of light is reduced, that is, the angle of the incident light 5 that satisfies the condition that the surface plasmon resonance occurs is measured.

Next, the solution 10 to be analyzed is subjected to dielectrophoresis.
A method for easily measuring the refractive index with high sensitivity when the concentration is low will be described.

A second metal thin film 12 and a third metal thin film 13 are held in addition to the first metal thin film 7 on the surface of the glass 6 in contact with the solution to be analyzed 10. Each of the metal thin films 12 and 13 is connected to an AC voltage source 14 and forms an electrode pair. Here, the metal thin films 12, 1
It is preferable that each of the surfaces of 3 is coated with a non-metallic substance, but it does not have to be coated.

Further, a temperature measuring probe 15 connected to a temperature measuring device 16 is provided in the immediate vicinity of the first metal thin film 7 for the purpose of correcting the temperature fluctuation of the solution 10 to be analyzed due to the fluctuation of the room temperature or the energization. However, a specific description regarding temperature correction will be given later in (Embodiment 2).

An AC voltage is applied between the pair of electrodes to exert a dielectrophoretic force on the molecules and particles contained in the solution to be analyzed 10 to generate molecules and particles on the first metal thin film 7 which produces surface plasmon resonance. Collect the particles. Since the electric dipole moment, that is, the easiness of gathering in the electric field concentrating portion, differs depending on the molecule or particle, the frequency or voltage value of the AC voltage may be set to an optimum value for each molecule or particle.

In order to make the dielectrophoretic force work, it is necessary to make the lines of electric force between the electrode pairs coarse and dense. An example of the electrode arrangement for that purpose is shown in FIG. Two metal thin films 12 forming electrodes,
Since the electric lines of force between 13 are most dense in the portion sandwiched by the ridges 17, if the first metal thin film 7 that causes surface plasmon resonance is provided here, molecules and particles in the solution 10 to be analyzed are provided. Will be collected on the first metal thin film 7 by the dielectrophoretic force.

By the way, in contrast to positive dielectrophoresis where electric lines of force gather in a dense place, there is also negative dielectrophoresis gathering in a rough place, but dielectrophoresis of molecules or particles having such characteristics is performed. In this case, the first metal thin film 7 may be provided in a place where the lines of electric force are rough.

By making the first metal thin film 7 function as an electrode, the number of metal thin films can be reduced. An example of the electrode arrangement for that purpose is shown in FIG. By using the circular container 11 and providing the second metal thin film 12 having an electrode function in the outer peripheral portion and the first metal thin film 18 having an electrode function in the central portion, the first metal thin film 18 is formed. The top is the densest place of the lines of electric force, and the molecules and particles in the solution to be analyzed 10 are gathered there.

By using the above-mentioned structure, the concentration of molecules and particles can be locally increased, and the refractive index can be measured by using surface plasmon resonance with higher sensitivity.
As a matter of course, when determining the refractive index of the solution to be analyzed 10 whose refractive index is unknown, the refractive index of the reference liquid having a known concentration is measured and compared with it.

When an unknown substance is detected, it is preferable to take out the collected molecules and particles and identify them by using a mass spectrometer such as TOF / MS.

Further, the light source 1, the photodetector 9, the AC voltage source 14, and the temperature measuring device 16 are each connected to a control arithmetic unit (not shown), and the procedure is preprogrammed or the operator is in a situation. Depending on the equipment control, measurement,
Detection, result, calculation, recording, etc. can be performed.

(Embodiment 2) In the present embodiment, in the refractive index measurement shown in (Embodiment 1), when the temperature of the first metal thin film 7 fluctuates, the influence on the surface plasmon resonance phenomenon is affected. A method of correction, in particular, a method of detecting the temperature of the first metal thin film 7 that causes the surface plasmon resonance phenomenon will be specifically described.

Although not shown in the drawings used in this embodiment, the optical system for measuring the surface plasmon resonance phenomenon and the electric circuit system for performing dielectrophoresis are those shown in FIG.

FIG. 4 is a schematic diagram showing temperature detection using a thermocouple. The hot junction 23 of the thermocouple 21 is connected to the first metal thin film 7
And the temperature fluctuations here are detected by the voltmeter 22 as fluctuations in the thermoelectromotive force. The voltmeter 22 is connected to a control calculation device (not shown), and the temperature of the first metal thin film 7 is calculated. To correct the temperature of the surface plasmon resonance phenomenon using the calculated temperature,
The temperature dependence of the surface plasmon resonance phenomenon is measured under known temperature conditions, and the temperature dependence is compared with that.

In this temperature detection method, since the temperature is detected at room temperature, chromel-alumel, platinum-platinum-rhodium, copper-constantan, etc. are preferably used as the metal constituting the thermocouple. In addition, the thermocouple 21
A fiber probe thermometer or the like may be used instead of the voltmeter 22. Furthermore, the temperature of the solution to be analyzed 10 may be obtained as long as it is in the immediate vicinity of the first metal thin film 7.

The first metal thin film 7 can also be made to function as a thermocouple. FIG. 5 is a schematic diagram showing temperature detection using the first metal thin film 7 as a thermocouple. The first metal wire 24 and the second metal wire 25 are electrically connected to the first metal thin film 7. When the first metal wire 24 and the first metal thin film 7 are made of the same kind of metal, the contact point between the second metal wire 25 and the first metal thin film 7 becomes the hot contact point 28 that generates thermoelectromotive force. Both metal wires 24 and 25 are connected to a conductor wire 26, and the conductor wire 26 is connected to a voltmeter 27. The thermoelectromotive force generated at the hot junction 28 is detected by the voltmeter 27. The contact points between the two metal wires 24 and 25 and the conductor wire 26 are cold contacts 29, which must be kept at a low temperature.

With the above-mentioned structure, the temperature of the hot junction 28, that is, the temperature of the first metal thin film 7 can be obtained by measuring the thermoelectromotive force generated at the hot junction 28.

Furthermore, it is also possible to determine the temperature variation of the metal from the variation of the electric resistance value of the metal due to the temperature variation. FIG. 6 is a schematic diagram showing the temperature detection using the electric resistance variation of the first metal thin film 7. The first metal thin film 7, constant voltage source 30, and ammeter 31 are connected by a conductor 32 to form a closed circuit. The electric resistance value of the first metal thin film 7 can be obtained from the voltage value indicated by the constant voltage source 30 and the current value indicated by the ammeter 31 according to Ohm's law.

Here, it is desirable that the conductive wire 32 be a metal whose electric resistance value has a small temperature fluctuation rate as compared with the first metal thin film 7. As a specific example, chromel or nichrome is used for the conductive wire 32, and C is used for the first metal thin film 7.
It is preferable to use o, Fe, Ni, Al, Au, Ag, Pt, Cu, W or the like.

Needless to say, the electric resistance value of the metal can be similarly obtained by using a voltmeter and a constant current source instead of the constant voltage source 30 and the ammeter 31.

As described above in detail, the temperature change of the solution to be analyzed 10 or the first metal thin film 7 is accurately measured, and the measurement result is corrected based on the temperature change, so that more accurate surface plasmon resonance can be obtained. The utilized refractive index measurement can be performed.

(Embodiment 3) In the present embodiment, in the refractive index measurement shown in (Embodiment 1), that is, in the refractive index measuring method shown in FIG. A refraction index measuring method that is not affected will be specifically described with reference to FIG.

FIG. 7 shows the incident angle at which the amount of reflected light is the smallest,
That is, it is a graph showing the time dependence of the incident angle θ satisfying the condition that surface plasmon resonance occurs. The vertical axis represents the resonance angle θ that satisfies the resonance generation condition, and the horizontal axis represents the time t. Further, the letters in the figure respectively represent a resonance angle θ1 when dielectrophoresis is not generated, a resonance angle θ2 immediately after dielectrophoresis is generated, a resonance angle θ3 immediately before dielectrophoresis is stopped, and a resonance angle θ3 immediately after dielectrophoresis is stopped. Resonance angle θ4, time t1 when dielectrophoresis is generated, and time t2 when dielectrophoresis is stopped.

Almost at the same time when dielectrophoresis is generated at time t1, molecules and particles in the solution to be analyzed 10 gather on the first metal thin film 7, and the resonance angle changes from θ1 to θ2.
From time t1 to t2, the resonance angle θ changes from θ2 to θ3 due to the temperature rise of the solution to be analyzed. When the dielectrophoresis is stopped at time t2, the molecules and particles on the first metal thin film 7 diffuse into the analyzed solution 10 due to Brownian motion, so that the resonance angle changes from θ3 to θ4 almost at the same time.

That is, if the resonance angle θ is measured by removing the variation from θ2 to θ3, it is possible to perform highly accurate refractive index measurement which is not affected by the temperature variation of the solution 10 to be analyzed. Specifically, the variation from θ1 to θ2 and the variation from θ3 to θ4 may be measured, and the refractive index may be measured from this value.

By using the above method, the refractive index measurement using the surface plasmon resonance with higher accuracy can be performed without being affected by the temperature fluctuations of the solution to be analyzed 10 and the first metal thin film 7. You can

However, since the easiness of gathering in the first metal thin film 7 and the easiness of diffusion thereof depend on the types of molecules and particles, it is not possible to apply the method described in this embodiment to all molecules and particles. Have difficulty. More specifically, for example, DNA molecules having a relatively large molecular weight are not instantaneously collected in the first metal thin film 7 at time t1, and it takes a time Δt until all of them are collected. If the time Δt is large, the temperature rise of the liquid 10 to be analyzed cannot be ignored, and t1
It is difficult to obtain an accurate refractive index from the value of the resonance angle θ2 ′ at + Δt.

In such a case, by using the temperature correction described in detail in (Embodiment 2) together, more accurate refractive index measurement using surface plasmon resonance can be performed.

(Embodiment 4) In the present embodiment, in the refractive index measurement shown in (Embodiment 1), that is, in the refractive index measurement method shown in FIG. The qualitative quantitative analysis method in the case of containing molecules and particles will be specifically described with reference to FIG.

FIG. 8 is a graph showing the time dependence of the migration distance L of a molecule or particle under the condition where dielectrophoresis is discretely generated. The vertical axis represents the moving distance L of the particle, and the horizontal axis represents the time t. The graph shown by a solid line in the figure shows the behavior of a relatively large molecule represented by DNA or the like, and the graph shown by a dotted line shows the behavior of a relatively small molecule represented by a protein. Further, the shaded region indicates a time zone in which no electric field is applied between the electrodes and dielectrophoresis does not occur.

In the time zone not covered by the diagonal lines, that is, in the time zone in which the dielectrophoresis is generated, the dielectrophoretic force acts on the molecule in the direction in which the electric field strength is strong and moves. The size of the moving distance is determined by the maximum electric field strength of the AC voltage, the frequency of the AC voltage, the electric field application time, the electric dipole moment peculiar to the molecule, etc. Generally, the larger molecule has a larger polarizability and the moving distance also increases. growing.

On the other hand, in the molecular behavior in the time zone covered by diagonal lines, that is, when dielectrophoresis is stopped, the external force acting on the molecule disappears, the Brownian motion becomes dominant, and the molecule diffuses. Contrary to the case of the dielectrophoretic force, the magnitude of the movement distance due to Brownian motion is larger for smaller molecules. When the stopping time is long, the small molecule returns to its original disordered state by Brownian motion.

As shown in FIG. 8, by alternately repeating the voltage application and the stop as described above, it is possible to collect only large molecules on the first metal thin film 7.

As described above, the types of molecules and particles collected by dielectrophoresis can be controlled by controlling the application time and the stop time of the AC voltage. By controlling the electric field strength and the frequency of alternating current in addition to the application / stop time, the size (mass) and shape of molecules and particles, polarizability,
Dielectric constant, resonance angle of surface plasmon resonance, etc. are used as parameters to analyze various kinds of molecules and particles by difference analysis of measurement results under different conditions and collection of molecules and particles collected in one place by dielectrophoresis. It is possible to perform a qualitative quantitative analysis of the solution to be analyzed.

Further, it is necessary to separate very similar molecules or particles which are difficult to separate only by parameters such as size (mass) or shape of molecules or particles, polarizability, permittivity, resonance angle of surface plasmon resonance. In this case, particles having clear distinctions in these parameters may be used as labels in advance, and qualitative and quantitative analysis of molecules or particles to be analyzed may be performed.

[0061]

As described above, according to the present invention, a simple, inexpensive refraction that is not affected by temperature fluctuations, does not require a complicated preparation process, and enables highly accurate and highly sensitive real-time measurement. Rate measurement can be realized.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing measurement of a refractive index of a solution to be analyzed according to an embodiment of the present invention.

FIG. 2 is a schematic diagram showing an electrode arrangement according to an embodiment of the present invention.

FIG. 3 is a schematic diagram showing an electrode arrangement according to an embodiment of the present invention.

FIG. 4 is a schematic diagram showing temperature detection using a thermocouple according to an embodiment of the present invention.

FIG. 5 is a schematic diagram showing temperature detection using a thermocouple according to an embodiment of the present invention.

FIG. 6 is a schematic diagram showing temperature detection using electric resistance variation according to an embodiment of the present invention.

FIG. 7 is a characteristic diagram showing time dependence of an incident angle θ according to an embodiment of the present invention.

FIG. 8 is a characteristic diagram showing the time dependence of the migration distance L of a molecule or particle according to an embodiment of the present invention.

[Explanation of symbols]

1 light source 2 first lens 3 Second lens 4 prism 5 incident light 6 glass 7 First metal thin film 8 reflected light 9 Photodetector 10 Analyte solution 11 containers 12 Second metal thin film 13 Third metal thin film 14 AC voltage source 15 Temperature measurement probe 16 Temperature measuring instrument 17 Ridge 18 First metal thin film 21 thermocouple 22 Voltmeter 23 Hot junction 24 First metal wire 25 Second metal wire 26 conductors 27 Voltmeter 28 hot junction 29 Cold junction 30 constant voltage source 31 ammeter 32 conductors

Claims (32)

[Claims] 1. A light source, a first metal thin film that causes surface plasmon resonance with light emitted from the light source, a photodetector that receives light reflected by the first metal thin film, and a solution to be analyzed by dielectrophoresis. An electrode pair consisting of a second metal thin film and a third metal thin film for, a voltage source for generating an alternating electric field between these electrodes, and a transparent substrate supporting these metal thin films,
A refractive index measuring device for a solution to be analyzed, comprising: a control calculation means for controlling a light source, a photodetector, a voltage source, etc., and calculating a refractive index from a measurement result obtained by the photodetector. 2. The refractive index measuring device according to claim 1, wherein the first metal thin film which causes surface plasmon resonance also has a function of an electrode for dielectrophoresing the solution to be analyzed. 3. Further, there is a correction means for correcting the refractive index of the solution to be analyzed and / or the metal thin film which has changed due to temperature change during the measurement of the refractive index, and the correction means has the solution to be analyzed and / or the metal thin film. 3. The refractive index measuring device according to claim 1, wherein the refractive index measuring device is configured by using the temperature detecting means of. 4. The refractive index measuring device according to claim 3, wherein the temperature detecting means obtains the temperature change of the metal thin film from the change of the electric resistance value of the metal thin film forming the electrode. 5. A light source, a metal thin film that causes surface plasmon resonance with light emitted from the light source, a photodetector that receives light reflected by the metal thin film, a transparent base material that supports the metal thin film, and an object to be analyzed. A temperature detecting means for the solution to be analyzed and / or the metal thin film for correcting the refractive index changed by the temperature change of the solution and / or the metal thin film, and a light source, a photodetector, a temperature detecting means, etc. An apparatus for measuring a refractive index of a solution to be analyzed, comprising: a control calculation means for calculating the refractive index of the solution to be analyzed from the obtained measurement result. 6. The refractive index measuring device according to claim 3, wherein the temperature detecting means is a temperature detecting device for directly detecting a temperature change of the solution to be analyzed and / or the metal thin film. 7. The refractive index measuring device according to claim 6, wherein the temperature detecting device is a thermocouple. 8. The temperature detecting means connects metal wiring made of a metal different from the metal thin film that causes surface plasmon resonance to the metal thin film, and measures the electromotive force generated at the interface between these different metals, The refractive index measuring device according to claim 3 or 5, wherein a temperature change of the thin film is obtained. 9. The method according to claim 1, wherein at least a portion in contact with the solution to be analyzed is covered with a non-metallic substance having a thickness of 1000 nm or less on the surface of the metal thin film which causes surface plasmon resonance. The refractive index measuring device according to any one of claims. 10. The surface of the metal thin film forming the electrode pair, at least a portion in contact with the solution to be analyzed is coated with a non-metal substance, according to any one of claims 1 to 4 or 6. The refractive index measuring device according to claim 8. 11. A surface plasmon resonance phenomenon generating means,
To measure the surface plasmon resonance phenomenon measuring means for measuring the change in the refractive index of the solution to be analyzed, the dielectrophoresis generating means for dielectrophoresing the solution to be analyzed, to control the surface plasmon resonance phenomenon, dielectrophoresis, etc., and to calculate the refractive index. And a voltage value and / or a voltage application time of an alternating electric field applied when dielectrophoresis is generated to control particles and molecules contained in the solution to be analyzed. A refractive index measuring method for measuring a refractive index in an assembled state. 12. A surface plasmon resonance phenomenon generating means,
Surface plasmon resonance phenomenon measuring means for measuring the change in the refractive index of the solution to be analyzed containing various kinds of particles and molecules,
A dielectrophoresis generating means for dielectrophores the solution to be analyzed and a control operation means for controlling the surface plasmon resonance phenomenon, dielectrophoresis, etc. and performing an operation to obtain the refractive index are provided, and dielectrophoresis is generated in the solution to be analyzed. At this time, by controlling the frequency and / or voltage value and / or voltage application time of the AC electric field to be applied, the types of particles or molecules trapped are controlled, and from the measurement results of the refractive index in each case,
A refractive index measuring method for obtaining a refractive index due to each of particles and molecules contained in a solution to be analyzed. 13. The refractive index measuring method according to claim 11, wherein the metal thin film that causes surface plasmon resonance also has a function of an electrode for dielectrophoresing the solution to be analyzed. 14. The method according to claim 11, further comprising a correction unit for correcting the influence of the temperature change of the solution to be analyzed and / or the metal thin film upon the refractive index measurement on the refractive index.
The method for measuring refractive index according to any one of 1. 15. In order to correct the influence of temperature change on the refractive index, the temperature change of the metal thin film is obtained from the change of the electric resistance value of the metal thin film forming the electrode for dielectrophoresing the solution to be analyzed. 15. The refractive index measuring method according to claim 14, further comprising a correcting unit that corrects based on the temperature change. 16. A correction means is provided which compares the refractive index before dielectrophoresis is generated with the refractive index immediately after dielectrophoresis is generated, thereby performing refractive index measurement without being affected by temperature change. The refractive index measuring method according to any one of claims 11 to 13. 17. A correction means for measuring a refractive index without being affected by a temperature change by comparing a refractive index before stopping dielectrophoresis and a refractive index immediately after stopping dielectrophoresis. The refractive index measuring method according to any one of claims 11 to 13. 18. Surface plasmon resonance phenomenon generating means,
Having a surface plasmon resonance phenomenon measuring means for measuring a change in the refractive index of the solution to be analyzed, a step of controlling the surface plasmon resonance phenomenon, a step of calculating the refractive index, and the solution to be analyzed at the time of measuring the refractive index and And / or a correction step for correcting the influence of the temperature change of the metal thin film on the refractive index. 19. In order to correct the influence of temperature change on the refractive index, a correction means is provided which directly measures the temperature change of the solution to be analyzed and / or the metal thin film and corrects based on the measured temperature change. The refractive index measuring method according to claim 14 or 18. 20. In order to correct the influence of temperature change on the refractive index, a metal wiring made of a metal different from the metal thin film that causes surface plasmon resonance is connected to the metal thin film, and this occurs at the interface between these dissimilar metals. 19. The refractive index measuring method according to claim 14 or 18, further comprising: a correction unit that obtains a temperature change of the metal thin film by measuring an electromotive force and corrects the temperature change based on the obtained temperature change. 21. A light source, a first metal thin film that causes surface plasmon resonance with light emitted from the light source, a photodetector that receives light reflected by the first metal thin film, and a solution to be analyzed by dielectrophoresis. Electrode pair consisting of a second metal thin film and a third metal thin film, a voltage source for generating an AC electric field between these electrodes, a transparent substrate supporting these metal thin films, a light source and a photodetector. And a control calculation device for calculating the refractive index from the measurement results obtained by controlling the voltage source and the photodetector, and the frequency of the AC voltage applied between the electrodes,
The voltage value of this AC voltage, the application time of this AC voltage,
From the refractive index obtained from the measurement results obtained with the photodetector,
A qualitative quantitative analyzer that identifies the type of molecules and particles in the solution to be analyzed and / or measures the number of molecules and particles in the solution to be analyzed. 22. The qualitative quantitative analysis device according to claim 21, wherein the first metal thin film which causes surface plasmon resonance also has a function of an electrode for dielectrophoresing the solution to be analyzed. 23. The correction means for correcting the refractive index changed by the temperature change of the solution to be analyzed and / or the metal thin film at the time of measuring the refractive index, the temperature detecting means of the solution to be analyzed and / or the metal thin film, and the control arithmetic unit. 23. The qualitative quantitative analysis device according to claim 21 or 22, which is constituted by 24. The qualitative quantitative analysis device according to claim 23, wherein the temperature detecting means obtains the temperature change of the metal thin film from the change of the electric resistance value of the metal thin film forming the electrode. 25. Compensating the refractive index before the dielectrophoresis is generated with the refractive index immediately after the dielectrophoresis is generated, so that the refractive index is measured without being affected by the temperature change. 23. The qualitative quantitative analysis device according to claim 21 or 22. 26. Compensating the refractive index before stopping dielectrophoresis and the refractive index immediately after stopping dielectrophoresis to provide a correction means for performing refractive index measurement without being affected by temperature change. 23. The qualitative quantitative analysis device according to claim 21 or 22. 27. A light source, a metal thin film that causes surface plasmon resonance with light emitted from the light source, a photodetector that receives light reflected by the metal thin film, a transparent base material that supports the metal thin film, and an object to be analyzed. A temperature detecting means for the solution to be analyzed and / or the metal thin film for correcting the refractive index changed by the temperature change of the solution and / or the metal thin film, and a light source, a photodetector, a temperature detecting means, etc. Equipped with a control calculation device for calculating the refractive index of the solution to be analyzed from the obtained measurement result, the molecules and particles in the solution to be analyzed from the refractive index obtained from the measurement result obtained by the photodetector A qualitative quantitative analysis device for identifying the type and / or measuring the number of molecules and particles in the solution to be analyzed. 28. The temperature detecting means is a solution to be analyzed and / or
28. The qualitative quantitative analysis device according to claim 23 or 27, which is a temperature measurement device for directly measuring a temperature change of the metal thin film. 29. The qualitative quantitative analysis device according to claim 28, wherein the temperature detection device is a thermocouple. 30. The temperature detecting means connects a metal wiring made of a metal different from the metal thin film that causes surface plasmon resonance to the metal thin film, and measures the electromotive force generated at the interface between these different metals, The qualitative and quantitative analysis device according to claim 23 or 27, which is to obtain a temperature change of the thin film. 31. The qualitative method according to any one of claims 21 to 30, wherein at least a portion in contact with the solution to be analyzed is covered with a non-metallic substance having a thickness of 1000 nm or less on the surface of the metal thin film which causes surface plasmon resonance. Quantitative analyzer. 32. The method according to any one of claims 21 to 26 or 28 to 30, wherein at least a portion of the surface of the metal thin film forming the electrode pair, which is in contact with the solution to be analyzed is covered with a non-metal substance. Qualitative quantitative analysis device described.