JP2008145369A - Sensing method, sensor for the same, and sensing apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To enhance a bond density of a labeling material to a sensor in a sensing method for detecting a to-be-detected material by labeling the to-be-detected material using the labeling material selectively bonded to the to-be-detected material if a sample liquid contains the to-be-detected material. <P>SOLUTION: The method includes and implements: a process (A) for bringing the sample liquid into contact with a sensing surface of the sensor; a process (B) for preparing a labeling material containing liquid containing the charged labeling material; a process (C) for applying a voltage to the labeling material containing liquid, and electrophoresing the charged labeling material within the labeling material containing liquid to the sensing surface in a state that the sample liquid is brought into contact with the sensing surface of the sensor; a process (F) for cleaning the sensing surface; and a process (G) for detecting a physical characteristic varied in response to the existence and the quantity of the charged labeling material bonded to the sensing surface. The sample liquid and the labeling material may be simultaneously brought into contact with the sensing surface of the sensor. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a sensing method for detecting a substance to be detected by labeling the substance to be detected with a labeling substance that selectively binds to the substance to be detected, a sensor suitable for use in implementing the method, and sensing Device.

  An immunochromatography method is known as an immunoassay method for detecting the presence and / or concentration of a substance to be detected by utilizing the specific binding property of an immune reaction (Patent Documents 1 and 2). The immunochromatography method is used in the field of qualitative analysis such as pregnancy test drugs and influenza virus antigen detection reagents.

  In general, in immunochromatography, a sensor having a sensing surface that is surface-modified so as to have a specific binding reaction with a substance to be detected is used, and the substance to be detected is labeled with a labeling substance that selectively binds to the substance to be detected, Sensing is performed by detecting the coloration caused by the labeling substance.

In conventional immunochromatography, gold particles are used as a labeling substance, and a method of detecting color development by localized plasmon scattering of gold particles, or a labeling substance that gives a specific color to a particle with a dye or the like is used. And a method of detecting the assigned color.
JP 2005-214670 A Japanese Patent Application No. 5-10950

  In the conventional immunochromatography method, the detected color is not so dark and the visual sensitivity is not so good. This is because the binding density of the substance to be detected and the labeling substance to the sensor used in the immunochromatography method is not sufficient.

The present invention has been made in view of such circumstances, and when a substance to be detected is labeled with a labeling substance that selectively binds to the substance to be detected to detect the substance to be detected, the substance to be detected is detected in a sample solution. An object of the present invention is to provide a sensing method capable of increasing the binding density of a labeling substance to a sensor when a substance is contained and capable of highly sensitive detection.
Another object of the present invention is to provide a sensor and a sensing device suitable for use in carrying out the sensing method described above.

The first sensing method of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In the sensing method for sensing,
A step (A) of bringing a sample liquid to be sensed into contact with the sensing surface of the sensor;
Preparing a labeling substance-containing liquid containing a chargeable labeling substance as the labeling substance (B);
In a state where the labeling substance-containing liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing liquid, and the chargeable labeling substance in the labeling substance-containing liquid is moved to the sensing surface side. Electrophoresing (C),
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that varies depending on the presence or absence or amount of the chargeable labeling substance bound on the sensing surface.

The second sensing method of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In the sensing method for sensing,
Preparing a labeling substance-containing sample liquid containing a chargeable labeling substance as the labeling substance for the sample liquid to be sensed (D);
In a state where the labeling substance-containing sample liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing sample liquid, and the chargeable labeling substance in the labeling substance-containing sample liquid is A step (E) of electrophoresis to the sensing surface side;
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that varies depending on the presence or absence or amount of the chargeable labeling substance bound on the sensing surface.

The first sensor of the present invention is the sensor used in the first or second sensing method of the present invention described above,
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
The plurality of micropores of the dielectric are filled with a conductive material, and the surface of the filled conductive material is subjected to surface modification capable of selectively binding to the detected material. It is a feature.

A second sensor of the present invention is the sensor used in the first or second sensing method of the present invention described above,
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
A surface in which the conductive material is filled in the plurality of micropores of the dielectric until it protrudes from the surface of the dielectric, and the surface of the filled conductive material can be selectively bonded to the detected material. It is characterized by being modified.

  The first and second sensors of the present invention preferably include a conductor that functions as an electrode when the chargeable labeling substance is electrophoresed on the sensing surface side on the back surface of the dielectric. .

The first sensing device of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In a sensing device that performs sensing,
The sensor, in which a labeling substance-containing liquid containing a chargeable labeling substance as the labeling substance is filled or flowing down, and the sample liquid that is the sensing target is previously brought into contact with the sensing surface, the sensing surface is the sensor A liquid cell attached so as to come into contact with the labeling substance-containing liquid;
Voltage applying means for applying a voltage to the labeling substance-containing liquid in the liquid cell and causing the chargeable labeling substance in the labeling substance-containing liquid to be electrophoresed on the sensing surface side of the sensor. It is characterized by this.

The second sensing device of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In a sensing device that performs sensing,
A sample substance-containing sample liquid containing a chargeable labeling substance as the labeling substance is filled or flowed into the sample liquid to be sensed, and the sensor has the sensing surface on the labeling substance-containing sample. A liquid cell attached to contact the liquid;
Voltage applying means for applying a voltage to the labeling substance-containing sample liquid in the liquid cell and causing the chargeable labeling substance in the labeling substance-containing sample liquid to be electrophoresed on the sensing surface side of the sensor; It is characterized by having.

In the first sensing method of the present invention, the sample liquid is brought into contact with the sensing surface of the sensor, and then the labeling substance-containing liquid containing the chargeable labeling substance is brought into contact therewith.
In the second sensing method of the present invention, a labeling substance-containing sample liquid containing a charging labeling substance is prepared for the sample liquid, and the sample liquid and the charging labeling substance are simultaneously brought into contact with the sensing surface of the sensor. Let
In any of the methods, a charged labeling substance is used as the labeling substance, and the process includes electrophoresis of the labeling substance to the sensing surface side using electrophoresis. In such a configuration, since the labeling substance can be effectively attracted to the sensing surface side by electrophoresis, it is possible to promote the binding between the labeling substance and the sensing surface when the target substance is contained in the sample solution. it can. As a result, it is possible to increase the binding density of the labeling substance to the sensor when the substance to be detected is contained in the sample solution, and highly sensitive detection is possible.

"Sensing method"
The first sensing method of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In the sensing method for sensing,
A step (A) of bringing a sample liquid to be sensed into contact with the sensing surface of the sensor;
Preparing a labeling substance-containing liquid containing a chargeable labeling substance as the labeling substance (B);
In a state where the labeling substance-containing liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing liquid, and the chargeable labeling substance in the labeling substance-containing liquid is moved to the sensing surface side. Electrophoresing (C),
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that varies depending on the presence or absence or amount of the chargeable labeling substance bound on the sensing surface.

  In the first sensing method of the present invention, the order of the step (A) and the step (B) may be any first. However, after the steps (A) and (B) are completed, the steps (C), (F), and (G) must be sequentially performed.

  In the step (B), as the “chargeable labeling substance”, a substance having originally chargeability may be used, or a substance having no chargeability may be used after being charged. When a charging process is performed on a substance having no charging property, the timing of the charging process is not particularly limited, and may be before or after preparing the labeling substance-containing liquid.

  Since the sensor used in the first sensing method of the present invention has a sensing surface capable of binding only a specific substance to be detected, if the substance to be detected is contained in the sample liquid, the process (A) In step (C), a substance to be detected is bonded to the sensing surface, and in step (C), a chargeable labeling substance is bonded to the substance to be detected bonded to the sensing surface. The charged labeling substance is not dropped off, and the charged labeling substance is detected in step (G). In the cleaning in the step (F), unnecessary chargeable labeling substances not bonded to the sensing surface are removed. If the sample liquid does not contain the substance to be detected, the charged labeling substance will not be bound to the sensing surface. Therefore, if the cleaning in the step (F) is performed, the charged labeling substance attached on the sensing surface will fall off. In step (G), no chargeable labeling substance is detected.

The second sensing method of the present invention uses a sensor having a sensing surface capable of binding only a specific substance to be detected, and labels the substance to be detected with a labeling substance that selectively binds to the substance to be detected. In the sensing method for sensing,
Preparing a labeling substance-containing sample liquid containing a chargeable labeling substance as the labeling substance for the sample liquid to be sensed (D);
In a state where the labeling substance-containing sample liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing sample liquid, and the chargeable labeling substance in the labeling substance-containing sample liquid is A step (E) of electrophoresis to the sensing surface side;
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that varies depending on the presence or absence or amount of the chargeable labeling substance bound on the sensing surface.

  In the second sensing method of the present invention, steps (D), (E), (F), and (G) need to be performed sequentially.

  In the step (D), as the “chargeable labeling substance”, a substance having originally chargeability may be used, or a substance having no chargeability may be used after being charged. When a charging process is performed on a substance having no charging property, the timing of the charging process is not particularly limited, and may be before or after preparing the labeling substance-containing sample solution.

  If the detected substance is contained in the sample liquid, in the step (D), a conjugate of the detected substance and the charged labeling substance is generated in the labeled substance-containing sample liquid. Since the sensor used in the second sensing method of the present invention has a sensing surface to which only a specific substance to be detected can bind, if the substance to be detected is contained in the sample liquid, the step (E) In FIG. 5, the conjugate of the substance to be detected and the chargeable labeling substance is bound to the sensing surface, and the chargeable labeling substance bound to the sensing surface does not fall off even after the cleaning in the step (F). ) To detect the charged labeling substance. In the cleaning in the step (F), unnecessary chargeable labeling substances not bonded to the sensing surface are removed. If the sample liquid does not contain the substance to be detected, the charged labeling substance will not be bound to the sensing surface. Therefore, if the cleaning in the step (F) is performed, the charged labeling substance attached on the sensing surface will fall off. In step (G), no chargeable labeling substance is detected.

  In the first and second sensing methods of the present invention, as the chargeable labeling substance, at least a metal particle having a surface made of metal, a fluorescent particle having at least a surface having fluorescence, and a colored particle having at least a surface colored It is preferable to use any one of the chargeable particles.

  In the first and second sensing methods of the present invention, the sensing surface is irradiated with measurement light, and changes depending on the presence or amount of the chargeable labeling substance on the sensing surface. It is preferable to detect the optical characteristics of the emitted light.

For example, when metal particles having at least a surface made of metal are used as the labeling substance, color development due to localized plasmon scattering can be detected.
For example, when fluorescent particles having at least a surface having fluorescence are used as the labeling substance, the fluorescent color can be detected.
For example, when colored particles having at least a surface colored are used as the labeling substance, the coloration can be detected.

In the first sensing method of the present invention, the sample liquid is brought into contact with the sensing surface of the sensor, and then the labeling substance-containing liquid containing the chargeable labeling substance is brought into contact therewith.
In the second sensing method of the present invention, a labeling substance-containing sample liquid containing a charging labeling substance is prepared for the sample liquid, and the sample liquid and the charging labeling substance are simultaneously brought into contact with the sensing surface of the sensor. Let
In any of the methods, a charged labeling substance is used as the labeling substance, and the process includes electrophoresis of the labeling substance to the sensing surface side using electrophoresis. In such a configuration, since the labeling substance can be effectively attracted to the sensing surface side by electrophoresis, it is possible to promote the binding between the labeling substance and the sensing surface when the target substance is contained in the sample solution. it can.

  In the first sensing method of the present invention, when a substance to be detected is contained in the sample liquid, the substance to be detected is first bound to the sensing surface of the sensor, and a labeling substance containing the labeling substance is bound thereto. Contact the containing liquid. In the present invention, since the labeling substance can be effectively attracted to the sensing surface side by electrophoresis, a larger number of labeling substances can be bound to many previously detected substances. As a result, it is possible to increase the binding density of the labeling substance to the sensor when the substance to be detected is contained in the sample solution, and highly sensitive detection is possible.

  In the second sensing method of the present invention in which the labeling substance is previously contained in the sample solution, when the detected substance is contained in the sample liquid, the detected substance and the labeled substance in the labeled substance-containing sample liquid A conjugate is generated. In the present invention, since the labeling substance can be effectively attracted to the sensing surface side by electrophoresis, the binding between the conjugate of the substance to be detected and the labeling substance and the sensing surface can be promoted. As a result, more conjugates of the substance to be detected and the labeling substance can be bound. As a result, it is possible to increase the binding density of the labeling substance to the sensor when the substance to be detected is contained in the sample solution, and highly sensitive detection is possible.

"Sensors and sensing devices"
With reference to the drawings, a configuration of a sensor and a sensing device according to an embodiment of the present invention will be described. FIG. 1 is an overall view of an apparatus according to an embodiment of the present invention, FIGS. 2A to 2D are diagrams showing the configuration of sensors according to the first to fourth embodiments of the present invention, and FIG. On the other hand, it is a figure which shows a mode that the to-be-detected substance and the labeling substance have couple | bonded.

  The sensing device 1 of the present embodiment uses a sensor 20 having a sensing surface 20s to which only a specific target substance R can be bound, and uses a labeling substance M that selectively binds the target substance R to the target substance R. A sensing device that performs labeling and sensing. In the present embodiment, a chargeable labeling substance is used as the labeling substance M.

  The sensing device 1 is disposed so as to be in contact with the liquid cell 10 in which a predetermined liquid 13 is filled or flows down, and the liquid 13 in the liquid cell 10, and the sensing surface 20s is irradiated with the measurement light L1. Sensor 20, a measuring light irradiation optical system 30 for irradiating the sensing surface 20s of the sensor 20 with the measuring light L1, and a detecting means 40 for detecting the optical characteristic of the outgoing light L2 from the sensing surface 20s.

  The liquid 13 filled or flowed into the liquid cell 10 includes a labeling substance-containing liquid used in the first sensing method of the present invention or a labeling substance used in the second sensing method of the present invention. A sample solution is used.

The measurement light irradiation optical system 30 is an optical system that irradiates the measurement surface L1 with respect to the sensing surface 20s, and introduces a light source 31, and a mirror, a lens, and the like that guide light emitted from the light source 31 as necessary. It is constituted by an optical system (not shown).
The measurement light L1 is not particularly limited, and may be single-wavelength light emitted from a light source such as a laser or broad light emitted from a white light source or the like, and is selected according to the detection method.

  The detection means 40 receives the outgoing light L2 including reflected light and scattered light generated on the sensing surface 20s of the sensor 20 by the irradiation of the measurement light L1, and depends on the presence or amount of the chargeable labeling substance M on the sensing surface 20s. It is a detector that detects the optical characteristics of the emitted light L2 that changes.

  The liquid cell 10 is a rectangular cell or the like having a bottom plate 11 and a top plate 12 that are spaced apart from each other. The liquid cell 10 is made of an insulating material. The upper and lower sides of the liquid cell 10 are determined for convenience, and the upper and lower sides of the liquid cell 10 can be designed as appropriate. In the present embodiment, the liquid cell 10 is provided with a voltage applying unit 50 that applies a voltage to the liquid 13 in the liquid cell 10.

  As shown in FIGS. 2A to 2D, in this embodiment, the sensor 20 includes an electrode 21, a dielectric 22 formed on the electrode 21 and having a plurality of micro holes 22a therein, and a plurality of It has a device structure composed of a conductive material 23 grown in the fine hole 22a.

  The sensor 20 is fixed to the liquid cell 10 with the electrode 21 fitted into the bottom plate 11 of the liquid cell 10 so that the sensing surface 20 s contacts the liquid 13 in the liquid cell 10. About the fixation aspect to the liquid cell 10 of the sensor 20, it can design suitably.

  The upper electrode 12 of the liquid cell 10 is fitted with a counter electrode 51 disposed in front of the sensor 20 via the liquid 13. A power source 52 and wiring 53 for applying a voltage to the electrode 21 and the counter electrode 51 that form a part of the sensor 20 are disposed outside the liquid cell 10. In the present embodiment, the voltage application means 50 is configured by the electrode 21, the counter electrode 51, the power source 52, and the wiring 53 that form part of the sensor 20.

With reference to FIG. 2, the suitable aspects 20A-20D of the sensor 20 are demonstrated.
As shown in FIGS. 4A to 4C, the sensor 20A shown in FIG. 2A shows the manufacturing process (FIGS. 4A and 4B are perspective views, and FIG. 4C is a sectional view). A part of the anodized metal body (Al, etc.) 60 is anodized to obtain a dielectric 22 made of a metal oxide (Al ₂ O ₃ etc.), and a plurality of dielectrics 22 formed in the process of anodization This is a device obtained by growing a conductive material 23 such as metal in each of the fine holes 22a by plating or the like. In this device, the conductive material 23 is filled in the fine holes 22 a of the dielectric 22 up to a position flush with the surface of the dielectric 22. In the sensor 20 </ b> A shown in FIG. 2A, the electrode 21 is made of a conductor made of a non-anodized portion (such as Al) of the anodized metal body 60.

  As shown in the figure, the dielectric 22 produced by anodic oxidation has a structure in which a plurality of substantially regular hexagonal fine columnar bodies 22b in a plan view are arranged without gaps, and is finely formed at a substantially central portion of each fine columnar body 22b. It has a structure in which the holes 22a are opened. Moreover, the bottom surface of each fine columnar body 22b has a rounded shape. The structure of the metal oxide body produced by anodization is Hideki Masuda, “Preparation of mesoporous alumina by anodization and application as a functional material”, Material Technology Vol.15, No.10, 1997, p.34, etc. It is described in. Since the fine holes 22a can be formed in a substantially regular pattern, the conductive materials 23 can be arranged in a substantially regular pattern.

  The sensor 20B shown in FIG. 2B has the same basic configuration as the sensor 20A shown in FIG. 2A, but the conductive material 23 is protruded from the surface of the dielectric 22 into the plurality of fine holes 22a. Is filled.

  The sensors 20C and 20D shown in FIGS. 2C and 2D are subjected to anodization in the same manner as in FIGS. 4A to 4C, and then the non-anodized portion of the metal body 60 to be anodized and its vicinity. The portion is removed to obtain the dielectric 22 in which the fine hole 22a is a through hole, and the electrode 21 is formed on the back side thereof. Thereafter, a conductive material such as metal is provided in each of the fine holes 22a of the dielectric 22 This is a device obtained by growing 23 by plating or the like. In the sensor 20C, the conductive material 23 is filled in the fine holes 22a of the dielectric 22 up to a position flush with the surface of the dielectric 22, and in the sensor 20D, the dielectric 22 is filled in the fine holes 22a of the dielectric 22. The conductive material 23 is filled until it protrudes from the surface. In the sensors 20C and 20D, the conductive material 23 and the electrode 21 are electrically connected.

  In the sensors 20A to 20D shown in FIGS. 2A to 2D, as shown in FIG. 3, the detection target substance R can be selectively bonded to the surface of the conductive substance 23 filled in the micro holes 22a. Surface modification A1 is applied, and the surface of the conductive material 23 is the sensing surface 20s (in FIG. 3, the sensor 20A is shown as a representative).

  For example, when the detected substance R is an antigen, as shown in FIG. 3, the surface of the conductive substance 23 is modified with a first antibody that specifically binds to the detected substance R (surface modification A1), and the labeling substance M is The surface may be modified with a second antibody that specifically binds to the substance to be detected R (surface modification A2). In this case, as shown in FIG. 3, the first antibody that is surface-modified to the conductive substance 23 and the second antibody that is surface-modified to the labeling substance M are mutually connected to the substance to be detected R that is an antigen. Those that bind to another site are used.

  In the sensors 20A to 20D shown in FIGS. 2A to 2D, the case of having the dielectric 22 formed by utilizing anodization has been described. However, the dielectric 22 having a plurality of fine holes 22a inside is described. A plurality of concave portions regularly arranged by a nanoimprint technique are formed on the surface of a substrate such as a resin. The surfaces of the substrate such as a metal are regularly arranged by an electron drawing technique such as a focused ion beam (FIB) or an electron beam (EB). It can also be obtained by a fine processing technique such as drawing a plurality of recesses. In this method, the fine holes 22a can be formed in an arbitrary pattern.

  The sensing device 1 of the present embodiment can be used when performing any one of the first sensing method and the second sensing method of the present invention.

  When the first sensing method of the present invention described above is performed, a sensing substance-containing liquid containing a chargeable labeling substance M is used as the liquid 13 filled or flowing down in the liquid cell 10, and sensing is performed on the sensing surface 20s in advance. The sensor 20 brought into contact with the target sample liquid is attached to the liquid cell 10, and a voltage is applied to the labeling substance-containing liquid 13 in the liquid cell 10 by the voltage applying means 50, so that the labeling substance-containing liquid 13 Is charged to the sensing surface 20 s side of the sensor 20.

  If the sample substance R is contained in the sample liquid, the substance R to be detected is bound to the sensing surface 20 s of the sensor 20 first, so that the charged labeling substance M in the labeling substance-containing liquid 13 is replaced with the sensor 20. As shown in FIG. 3, the charged labeling substance M binds to the substance R to be detected bound to the sensing face 20s.

  When the second sensing method of the present invention described above is carried out, a labeling substance-containing sample liquid in which the charged labeling substance M is contained in the sample liquid is used as the liquid 13 that fills or flows down in the liquid cell 10. The sensor 20 that is used and not pretreated is attached to the liquid cell 10, and a voltage is applied to the labeling substance-containing sample liquid 13 in the liquid cell 10 by the voltage application means 50, thereby the labeling substance-containing sample liquid 13. The charged labeling substance M therein is electrophoresed on the sensing surface 20 s side of the sensor 20.

  If the sample substance R is contained in the sample liquid, since the conjugate of the substance R to be detected and the chargeable label substance M has been generated in the sample substance-containing sample liquid 13, the label substance-containing sample When the chargeable labeling substance M in the liquid 13 is electrophoresed on the sensing surface 20s side of the sensor 20, the conjugate of the substance R to be detected and the chargeable labeling substance M generated in the sample substance-containing sample liquid becomes the sensing surface 20s. To join.

  In any case, if the electrode 21 is an anode, the labeling substance M is adjusted to a negatively charged state, and if the electrode 21 is a cathode, the labeling substance M is adjusted to a positively charged state. The labeling substance M can be attracted to the sensing surface 20s side.

  In any of the methods, the sensing cell 20 s is washed on the sensing surface 20 s after the inside of the liquid cell 10 after electrophoresis of the labeling substance M and filled with a liquid that does not hinder measurement such as water. On the other hand, the measurement light L1 is irradiated and the emission light L2 is detected. Since the example of the combination of the type of the labeling substance M and the optical property to be detected has been described above, it will be omitted.

  By using the sensing device 1 of the present embodiment, the above first sensing method of the present invention or the above second sensing method of the present invention can be implemented. By using the sensing device 1 of the present embodiment, the binding density of the labeling substance M to the sensor 20 when the sample substance R is contained in the sample liquid can be increased, and highly sensitive detection is possible. .

  In the present embodiment, the sensor 20 includes an electrode 21, a dielectric 22 formed on the electrode 21 and having a plurality of micro holes 22a therein, and a conductive material 23 grown in the plurality of micro holes 22a. The case where the device structure is as described above has been described.

  As the sensor 20, a simple metal plate (with or without surface irregularities) to which the surface modification A1 is applied can be used.

  However, the sensor 20 having the configuration described in this embodiment is preferably used for the following reason. The sensor 20 described in the present embodiment has a structure in which a plurality of conductive materials 23 to which a surface modification A1 that selectively binds to a detected substance R is applied are isolated from each other through a fine columnar body 22b. Yes. In such a configuration, it is possible to prevent the conjugate of the substance R to be detected and the labeling substance M from being aggregated at a certain location on the sensing surface 20s, and to detect the substance R to be detected and the labeling substance on the sensing surface 20s. A combination with M can be arranged neatly, and sensing can be performed uniformly over the entire sensing surface 20s, which is preferable.

  The technology of the present invention can be preferably used for biosensing or the like in which a substance to be detected is labeled with a labeling substance that selectively binds to the substance to be detected and the substance to be detected is detected.

1 is an overall view of a sensing device according to an embodiment of the present invention. (A)-(d) is a figure which shows the structure of the sensor of 1st-4th embodiment which concerns on this invention. The figure which shows a mode that the to-be-detected substance and the labeling substance have couple | bonded with respect to the sensor. (A)-(c) is a manufacturing process figure of the sensor shown to Fig.2 (a).

Explanation of symbols

1 Sensing device 10 Liquid cell 13 Liquid in liquid cell (labeling substance-containing liquid or labeling substance-containing sample liquid)
20, 20A to 20D Sensor 20s Sensing surface 21 Electrode (conductor)
22 Dielectric 22a Micropore 23 Conductive substance 30 Measurement light irradiation optical system 40 Detection means 50 Voltage application means 51 Counter electrode L1 Measurement light L2 Emission light R Detected substance M Chargeable labeling substance A1, A2 Surface modification

Claims (14)

Using a sensor with a sensing surface that can bind only specific substances to be detected,
In the sensing method of sensing by labeling the detected substance with a labeling substance that selectively binds to the detected substance,
A step (A) of bringing a sample liquid to be sensed into contact with the sensing surface of the sensor;
Preparing a labeling substance-containing liquid containing a chargeable labeling substance as the labeling substance (B);
In a state where the labeling substance-containing liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing liquid, and the chargeable labeling substance in the labeling substance-containing liquid is moved to the sensing surface side. Electrophoresing (C),
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that changes depending on the presence or amount of the chargeable labeling substance bound on the sensing surface. Using a sensor with a sensing surface that can bind only specific substances to be detected,
In the sensing method of sensing by labeling the detected substance with a labeling substance that selectively binds to the detected substance,
Preparing a labeling substance-containing sample liquid containing a chargeable labeling substance as the labeling substance for the sample liquid to be sensed (D);
In a state where the labeling substance-containing sample liquid is in contact with the sensing surface of the sensor, a voltage is applied to the labeling substance-containing sample liquid, and the chargeable labeling substance in the labeling substance-containing sample liquid is A step (E) of electrophoresis to the sensing surface side;
A step (F) of cleaning the sensing surface;
And (G) detecting a physical property that changes depending on the presence or amount of the chargeable labeling substance bound on the sensing surface.   The chargeable labeling substance is characterized by using a chargeable particle comprising at least one of metal particles having a surface made of metal, at least a fluorescent particle having a surface having fluorescence, and a colored particle having at least a surface colored. The sensing method according to claim 1 or 2.   Irradiating measurement light to the sensing surface, and detecting optical characteristics of light emitted from the sensing surface, which changes depending on the presence or amount of the chargeable labeling substance on the sensing surface. The sensing method in any one of Claims 1-3. The sensor used in the sensing method according to any one of claims 1 to 4,
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
The plurality of micropores of the dielectric are filled with a conductive material, and the surface of the filled conductive material is subjected to surface modification capable of selectively binding to the detected material. A featured sensor. The sensor used in the sensing method according to any one of claims 1 to 4,
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
A surface in which the conductive material is filled in the plurality of micropores of the dielectric until it protrudes from the surface of the dielectric, and the surface of the filled conductive material can be selectively bonded to the detected material. A sensor characterized by being modified.   7. The sensor according to claim 5, wherein a conductor functioning as an electrode when the chargeable labeling substance is electrophoresed on the sensing surface side is provided on the back surface of the dielectric. Using a sensor with a sensing surface that can bind only specific substances to be detected,
In a sensing device that performs sensing by labeling the detected substance with a labeling substance that selectively binds to the detected substance,
The sensor, in which a labeling substance-containing liquid containing a chargeable labeling substance as the labeling substance is filled or flowing down, and the sample liquid that is the sensing target is previously brought into contact with the sensing surface, the sensing surface is the sensor A liquid cell attached so as to come into contact with the labeling substance-containing liquid;
Voltage applying means for applying a voltage to the labeling substance-containing liquid in the liquid cell and causing the chargeable labeling substance in the labeling substance-containing liquid to be electrophoresed on the sensing surface side of the sensor. Sensing device characterized by that. Using a sensor with a sensing surface that can bind only specific substances to be detected,
In a sensing device that performs sensing by labeling the detected substance with a labeling substance that selectively binds to the detected substance,
A sample substance-containing sample liquid containing a chargeable labeling substance as the labeling substance is filled or flowed into the sample liquid to be sensed, and the sensor has the sensing surface on the labeling substance-containing sample. A liquid cell attached to contact the liquid;
Voltage applying means for applying a voltage to the labeling substance-containing sample liquid in the liquid cell and causing the chargeable labeling substance in the labeling substance-containing sample liquid to be electrophoresed on the sensing surface side of the sensor; A sensing device characterized by comprising.   Measuring light irradiation optical system for irradiating the sensing surface of the sensor with the measuring light, and optical characteristics of light emitted from the sensing surface, which changes depending on the presence or amount of the chargeable labeling substance on the sensing surface The sensing device according to claim 8, further comprising a detecting unit that detects s. The sensor is
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
The plurality of micropores of the dielectric are filled with a conductive material, and the surface of the filled conductive material is subjected to surface modification capable of selectively binding to the detected material. The sensing device according to any one of claims 8 to 10. The sensor is
A dielectric having a plurality of micropores therein and having at least a plurality of micropores open on the surface on the sensing surface side;
A surface in which the conductive material is filled in the plurality of micropores of the dielectric until it protrudes from the surface of the dielectric, and the surface of the filled conductive material can be selectively bonded to the detected material. The sensing device according to any one of claims 8 to 10, wherein the sensing device is modified.   The sensing device according to claim 11 or 12, wherein the sensor includes a conductor on a back surface of the dielectric.   14. The sensing according to claim 13, wherein the voltage applying means includes an electrode made of the conductor provided in the sensor and a counter electrode disposed to face the sensor. apparatus.

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