Micro devices and analytical devices

Abstract

A microdevice and analytical apparatus are provided. The microdevice includes: a microchannel into which a target solution containing a target substance is introduced; an antibody immobilized to at least one sidewall surface of the microchannel and specifically binding to the target substance; a fluorescently labeled derivative specifically binding to the antibody and obtained by fluorescently labeling the target substance; and a light blocker blocking excitation light that excites fluorescence emitted by the fluorescently labeled derivative. The target substance and the fluorescently labeled derivative bind specifically to the antibody in a competitive manner, and the antibody is immobilized to the sidewall surface of the microchannel in a state of specific binding to the fluorescently labeled derivative. The light blocker blocks excitation light entering the fluorescently labeled derivative specifically bound to the antibody.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of Japanese Patent Application No. 2020-128025, filed on July 29, 2020, the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] This application generally relates to micro-devices and analytical devices. Background Technology

[0004] Fluorescence polarization immunoassay (FPIA), which measures a target substance by detecting an antigen-antibody reaction, is called fluorescence immunoassay. For example, unexamined Japanese Patent Application Publication No. H3-103765 ​​discloses a method for determining the concentration of the antigen (target substance) being measured based on the polarization of the measured fluorescence.

[0005] Furthermore, methods for generating antigen-antibody reactions in microchannels within microdevices are known. By generating antigen-antibody reactions in microchannels, the reaction time of antigen-antibody reactions can be shortened. For example, Japanese Patent No. 4717081 discloses an immunoassay microchip in which microstructures are placed in channels. The microstructures contain beads, and primary antibodies are immobilized on the surface of the beads.

[0006] For example, in a fluorescence polarization immunoassay using conventional micro-devices, a solution containing the target substance and a fluorescently labeled derivative obtained by labeling the target substance with a fluorescent agent is introduced into a channel to which an antibody is immobilized. After the competitive reaction (antigen-antibody reaction) between the target substance and the fluorescently labeled derivative reaches equilibrium, the solution in the channel is irradiated with linearly polarized excitation light that excites fluorescence emitted by the fluorescently labeled derivative, and the degree of polarization of the fluorescence radiated from the solution is determined. The degree of polarization of the fluorescence varies with the concentration of the target substance, and therefore the concentration of the target substance can be obtained from a previously generated calibration curve.

[0007] In fluorescence polarization immunoassays using conventional micro-devices, the fluorescence emitted from the solution includes fluorescence emitted by fluorescently labeled derivatives that specifically bind to the antibody and fluorescence emitted by fluorescently labeled derivatives that do not bind to the antibody, resulting in low measurement sensitivity.

[0008] This disclosure is made in view of the foregoing circumstances, and the purpose of this disclosure is to provide a miniature device and analytical apparatus with high measurement sensitivity to the target substance. Summary of the Invention

[0009] To achieve the aforementioned objectives, the micro-device according to the first aspect of this disclosure includes:

[0010] A microchannel, into which a target solution containing the target substance is introduced;

[0011] An antibody is immobilized on at least one sidewall surface of the microchannel and specifically binds to the target substance being measured.

[0012] A fluorescently labeled derivative, which specifically binds to an antibody and is obtained by measuring the target substance via fluorescent labeling; and

[0013] A light blocker that blocks the excitation light that excites fluorescence emitted by a fluorescently labeled derivative, wherein...

[0014] The target substance and its fluorescently labeled derivatives are measured to bind specifically to the antibody in a competitive manner.

[0015] The antibody was immobilized on the sidewall surface of the microchannel in a state of specific binding to the fluorescently labeled derivative, and

[0016] The light blocker blocks the excitation light from entering the fluorescently labeled derivative that specifically binds to the antibody.

[0017] The micro device according to the second aspect of this disclosure includes:

[0018] A microchannel containing a target substance for measurement and a target solution containing a fluorescently labeled derivative obtained by fluorescent labeling the target substance is introduced into the microchannel;

[0019] An antibody, immobilized on at least one sidewall surface of a microchannel, specifically binds to the target substance being measured and a fluorescently labeled derivative; and

[0020] A light blocker that blocks the excitation light that excites fluorescence emitted by a fluorescently labeled derivative, wherein...

[0021] The target substance and fluorescently labeled derivatives are measured to bind specifically to the antibody in a competitive manner, and,

[0022] When the target solution is introduced into the microchannel, the light blocker blocks the excitation light from entering the fluorescently labeled derivative that specifically binds to the antibody.

[0023] The analytical apparatus according to the third aspect of this disclosure includes:

[0024] One of the aforementioned micro-devices;

[0025] A radiator that utilizes a micro-device to emit excitation light; and

[0026] The detector detects fluorescence.

[0027] It should be understood that the foregoing overview and the following detailed description are exemplary and explanatory, and do not limit this disclosure.

[0028] This disclosure can improve the measurement sensitivity of the target substance. Attached Figure Description

[0029] A more complete understanding of this application can be obtained by considering the following detailed description in conjunction with the accompanying drawings, in which:

[0030] Figure 1 This is a top view showing the micro device according to Embodiment 1;

[0031] Figure 2 yes Figure 1 The cross-sectional view of the micro device taken along line AA is shown in the figure;

[0032] Figure 3 This is a schematic diagram illustrating the antibody and fluorescently labeled derivative according to Example 1;

[0033] Figure 4 This is a schematic diagram illustrating the operation of the microdevice according to Embodiment 1;

[0034] Figure 5 This is a flowchart illustrating a method for manufacturing a microdevice according to Embodiment 1;

[0035] Figure 6 This is a schematic diagram illustrating the process of integrally forming the second substrate and the light blocker according to Embodiment 1;

[0036] Figure 7 This is a diagram showing the configuration of the analysis device according to Embodiment 1;

[0037] Figure 8 This is a schematic diagram showing the analysis apparatus according to Embodiment 1;

[0038] Figure 9 This is a top view showing the micro device according to Embodiment 2;

[0039] Figure 10 yes Figure 9 The image shows a cross-sectional view of the micro device taken along line BB;

[0040] Figure 11 This is a schematic diagram illustrating the antibody and fluorescently labeled derivative according to Example 2;

[0041] Figure 12 This is a top view showing the micro device according to Embodiment 3;

[0042] Figure 13 This is a schematic diagram showing the sidewall surface of the microchannel in the radiation region according to Embodiment 3;

[0043] Figure 14 yes Figure 12 The image shows a cross-sectional view of the micro device taken along line CC;

[0044] Figure 15 yes Figure 12 The image shows a cross-sectional view of the micro device taken along line DD;

[0045] Figure 16 yes Figure 12 The cross-sectional view of the micro device taken along line EE is shown in the figure;

[0046] Figure 17 This is a top view showing the fourth light blocker and the second substrate according to Embodiment 3;

[0047] Figure 18 This is a flowchart illustrating a method for manufacturing a microdevice according to Embodiment 3;

[0048] Figure 19 This is a schematic diagram according to Example 3 illustrating the process of immobilizing an antibody to the side of a third light blocker and specifically binding a fluorescently labeled derivative to the immobilized antibody.

[0049] Figure 20 This is a schematic diagram showing the antibody according to Example 4;

[0050] Figure 21 This is a schematic diagram showing a cross-section of a micro device based on a modified example;

[0051] Figure 22 This is a schematic diagram showing a cross-section of a mold based on a modified example;

[0052] Figure 23 This is a schematic diagram illustrating an analysis device based on a modified example; and

[0053] Figure 24 This is a schematic diagram showing a cross-section of a microdevice based on a modified example. Detailed Implementation

[0054] The miniature device according to an embodiment will be described below with reference to the accompanying drawings.

[0055] Example 1

[0056] Reference Figures 1 to 8 The microdevice 10 according to this embodiment is described. For example, the microdevice 10 is used to detect and measure the target substance Ag1 by using fluorescence polarization immunoassay.

[0057] like Figure 1 and Figure 2 As shown, the microdevice 10 includes a first substrate 12, a second substrate 14, a first light blocker 16, and three microchannels 18. Additionally, as... Figure 3 As shown, the microdevice 10 includes an antibody Ab1 and a fluorescently labeled derivative AgF1. The fluorescently labeled derivative AgF1 is a derivative obtained by using a fluorescent substance to fluorescently label the target substance Ag1 for measurement.

[0058] The first substrate 12 and the second substrate 14 sandwich the first light blocker 16 between them. The first light blocker 16 blocks the excitation light EL emitted by the fluorescently labeled derivative AgF1, which is excited by fluorescence FL. Further, the first substrate 12, the second substrate 14, and the first light blocker 16 form a microchannel 18. A measurement target solution containing the measurement target substance Ag1 is introduced into the microchannel 18. Antibody Ab1 is immobilized on the sidewall surface 18a of the microchannel 18. The fluorescently labeled derivative AgF1 specifically binds to antibody Ab1.

[0059] For ease of understanding, Figure 1 The rightward direction (towards the right side of the page) of the microdevice 10 is represented herein as the +X direction, the upward direction (towards the top of the page) as the +Y direction, and the direction perpendicular to the +X and +Y directions (towards the back of the page) as the +Z direction. Furthermore, antibody Ab1 and the fluorescently labeled derivative AgF1... Figure 1 and Figure 2 The middle part is omitted.

[0060] The first substrate 12 in the microdevice 10 is a plate-shaped silicon dioxide glass substrate. For example... Figure 2 As shown, the excitation light EL enters the microdevice 10 from the first substrate 12.

[0061] The excitation light EL is the light that excites the fluorescent FL emitted by the fluorescently labeled derivative AgF1. In this embodiment, the excitation light EL radiates... Figure 1 The radiation region S is shown in the figure. The excitation light EL enters perpendicularly to the first principal plane 12a of the first substrate 12.

[0062] The second substrate 14 in the microdevice 10 is a plate-shaped substrate. The second substrate 14 is formed of a material with low autofluorescence. In this embodiment, the second substrate 14 is formed of polydimethylsiloxane (PDMS) containing carbon black. The second substrate 14 faces the first substrate 12. The second substrate 14 and the first substrate 12 sandwich the first light blocker 16 between them.

[0063] The first light blocker 16 in the microdevice 10 forms a microchannel 18 by being sandwiched between the first substrate 12 and the second substrate 14, and blocks the excitation light EL entering from the first substrate 12. The first light blocker 16 absorbs the excitation light EL that excites the fluorescence FL emitted by the fluorescently labeled derivative AgF1, and is formed of a material with low autofluorescence. In this embodiment, the first light blocker 16 is integrally formed of polydimethylsiloxane containing carbon black with the second substrate 14.

[0064] When viewed in a cross-section (YZ plane) along the width direction of the microchannel 18, the side portion 16a of the first light blocker 16 forms the sidewall surface 18a of the microchannel 18. In this embodiment, the two side portions 16a forming the two sidewall surfaces 18a of the microchannel 18 are inclined in a direction that narrows the width of the microchannel 18 (i.e., its length in the Y direction) toward the first substrate 12. In other words, the two side portions 16a are inclined in a direction that brings them closer to each other toward the first substrate 12. Figure 3 As shown, antibody Ab1 is immobilized on the side 16a of the first light blocker 16 (i.e., the sidewall surface 18a of the microchannel 18) in a state that specifically binds to the fluorescently labeled derivative AgF1.

[0065] In this embodiment, the side portion 16a of the first light blocker 16 is inclined in a direction that narrows the width of the microchannel 18 toward the first substrate 12, and the excitation light EL enters from the first substrate 12 perpendicular to the first main plane 12a. Therefore, as Figure 3 As shown, the first light blocker 16 blocks the excitation light EL of the fluorescently labeled derivative AgF1, which specifically binds to the antibody Ab1 immobilized on the side 16a (i.e., the sidewall surface 18a of the microchannel 18). For ease of understanding, the fluorescently labeled derivative that specifically binds to the antibody immobilized on the sidewall surface 18a can be described below as the fluorescently labeled derivative that is bound.

[0066] Microchannels 18 in the microdevice 10 extend parallel to each other in the X direction within the radiation region S. A measurement target solution containing the target substance Ag1 is introduced into the microchannels 18. The target substance Ag1 only needs to be a compound that can be detected by using a fluorescent immunoassay. Examples of target substances Ag1 include antibiotics, physiologically active substances, and fungal toxins. Specific examples of target substances Ag1 include prostaglandin E2, β-lactoglobulin, chloramphenicol, and deoxynivalenol. The measurement target solution containing the target substance Ag1 is introduced into the microchannels 18 from inlet 19a. Inlet 19a passes through the second substrate 14 and the first light blocker 16 and is connected to the microchannels 18. Further, the measurement target solution within the microchannels 18 is discharged from outlet 19b. Outlet 19b passes through the second substrate 14 and the first light blocker 16 and is connected to the microchannels 18. For ease of understanding, the measurement target solution containing the target substance can be described hereinafter as a measurement target solution.

[0067] Microchannel 18 is formed by a first substrate 12, a second substrate 14, and a first light blocker 16. The upper wall surface 18b of microchannel 18 is formed by the second principal plane 12b of the first substrate 12. The lower wall surface 18c of microchannel 18 is formed by the first principal plane 14a of the second substrate 14. The sidewall surfaces 18a of microchannel 18 are formed by the side portions 16a of the first light blocker 16. The side portions 16a of the first light blocker 16 are inclined in a direction that narrows the width of microchannel 18 towards the first substrate 12, and therefore both sidewall surfaces 18a of microchannel 18 are also inclined in a direction that narrows the width of microchannel 18 towards the first substrate 12. In other words, the two sidewall surfaces 18a are inclined in a direction that brings them closer to each other towards the first substrate 12. Figure 3 As shown, antibody Ab1 is immobilized on the sidewall surface 18a of the microchannel 18 (i.e., the side 16a of the first light blocker 16).

[0068] In this embodiment, the sidewall surface 18a is inclined in a direction that narrows the width of the microchannel 18 toward the first substrate 12, and therefore the cross-section of the microchannel 18 in the width direction (Y direction) has a tapered shape that narrows toward the first substrate 12. For example, the width of the microchannel 18 is 210 μm at its widest point and 50 μm at its narrowest point. For example, the depth (length in the Z direction) of the microchannel 18 is 900 μm. Further, the ratio of the depth of the microchannel 18 to its narrowest width is preferably 2 or greater.

[0069] In the microdevice 10, antibody Ab1 is immobilized on the sidewall surface 18a of the microchannel 18 (i.e., the side 16a of the first light blocker 16). In this embodiment, antibody Ab1 is immobilized on the sidewall surface 18a of the microchannel 18 in a state specifically bound to the fluorescently labeled derivative AgF1. Antibody Ab1 specifically binds to the target substance Ag1 through an antigen-antibody reaction. For example, antibody Ab1 is obtained by inoculating the target substance Ag1 into a host animal (such as a mouse or cow) and then collecting and purifying the antibodies in the blood produced by the host animal. Alternatively, commercially available antibodies can also be used as antibody Ab1.

[0070] The fluorescently labeled derivative AgF1 in the microdevice 10 specifically binds to antibody Ab1 via an antigen-antibody reaction. The fluorescently labeled derivative AgF1 is obtained by binding a fluorescent substance to the target substance Ag1 using a method generally known to be used. Examples of fluorescent substances include fluorescein (excitation wavelength EL: 494 nm, fluorescence wavelength FL: 521 nm) and rhodamine β (excitation wavelength EL: 550 nm, fluorescence wavelength FL: 580 nm). In this embodiment, the first light blocker 16 blocks the excitation light EL, and therefore the fluorescently labeled derivative AgF1 (the bound fluorescently labeled derivative AgF1) specifically bound to the antibody Ab1 fixed to the sidewall surface 18a does not fluoresce even when the microdevice 10 is irradiated by the excitation light EL.

[0071] The operation of the microdevice 10 will be described.

[0072] When the target solution is introduced into the microchannel 18 of the microdevice 10, the target substance Ag1 contained in the target solution specifically binds to antibody Ab1 through an antigen-antibody reaction that competes with the fluorescently labeled derivative AgF1. Then, as... Figure 4 As shown, a fluorescently labeled derivative AgF1 that does not specifically bind to antibody Ab1 is generated in microchannel 18. When the competitive reaction between the target substance Ag1 and the fluorescently labeled derivative AgF1 against antibody Ab1 reaches equilibrium, the fluorescently labeled derivative AgF1, in an amount determined according to the concentration of the target substance Ag1, is generated in microchannel 18. For ease of understanding, the fluorescently labeled derivative that does not specifically bind to the antibody may be described hereinafter as a free fluorescently labeled derivative.

[0073] When the competitive reaction reaches equilibrium, and excitation light EL enters from the first substrate 12, the first light blocker 16 blocks the excitation light EL from the bound fluorescently labeled derivative AgF1, thus suppressing the fluorescence emitted by the bound fluorescently labeled derivative AgF1. The fluorescence emitted by the free fluorescently labeled derivative AgF1 is then measured. Therefore, the microdevice 10 can improve the measurement sensitivity of the target substance Ag1.

[0074] Next, we will refer to Figure 5 and Figure 6 A method for manufacturing the micro-device 10 is described. Figure 5 This is a flowchart illustrating a method for manufacturing the microdevice 10. The method for manufacturing the microdevice 10 includes: a process of integrally forming a second substrate 14 and a first light blocker 16 (step S10), a process of forming an inlet 19a and an outlet 19b (step S20), a process of bonding the first light blocker 16 to a first substrate 12 (step S30), a process of immobilizing antibody Ab1 to the sidewall surface 18a of a microchannel 18 (step S40), and a process of specifically binding a fluorescently labeled derivative AgF1 to antibody Ab1 (step S50).

[0075] In step S10, a mold 62, whose shape is suitable for the second substrate 14 and the first light blocker 16, is placed in a template 64, such as... Figure 6 As shown. Then, polydimethylsiloxane containing carbon black is poured into template 64. By curing the polydimethylsiloxane poured into template 64, the second substrate 14 and the first light blocker 16 are integrally formed. For example, the mold 62 is produced by photolithography on a silicon substrate. Further, the component that forms the integral part of the second substrate 14 and the first light blocker 16 can be described below as including the second substrate 14, which contains the first light blocker 16.

[0076] Back Figure 5 In step S20, an inlet 19a and an outlet 19b are formed by using a fixture to create through holes at predetermined positions on a second substrate 14 including the first light blocker 16.

[0077] In step S30, the first light blocker 16 is bonded to the first substrate 12 by placing the first substrate 12 on the first light blocker 16 and then pressing the first substrate 12 against the first light blocker 16. Thus, the first substrate 12, the second substrate 14, and the first light blocker 16 form a microchannel 18.

[0078] In step S40, a solution containing antibody Ab1 is introduced into the microchannel 18, and antibody Ab1 is immobilized onto the sidewall surface 18a of the microchannel 18 (side 16a of the first light blocker 16). After antibody Ab1 is immobilized onto the sidewall surface 18a of the microchannel 18, a predetermined solution is introduced into the microchannel 18, and the interior of the microchannel 18 is cleaned. Depending on the characteristics of antibody Ab1, methods for immobilizing antibody Ab1 may include physical adsorption, covalently binding or ionically binding antibody Ab1 to the sidewall surface 18a, or other commonly known methods.

[0079] In step S50, a solution containing a fluorescently labeled derivative AgF1 is introduced into microchannel 18, and the fluorescently labeled derivative AgF1 specifically binds to antibody Ab1 via an antigen-antibody reaction. After the fluorescently labeled derivative AgF1 binds to antibody Ab1, a predetermined solution is introduced into microchannel 18, and the interior of microchannel 18 is cleaned. Thus, microdevice 10 can be generated.

[0080] The detection of the target substance Ag1 using a micro-device 10 will be described. First, the analytical apparatus 100 for detecting the target substance Ag1 will be described.

[0081] like Figure 7 and Figure 8 As shown, the analysis device 100 includes a radiator 110, a dichroic mirror 120, an objective lens 130, a detector 140, and a controller 150. The analysis device 100 also includes a microdevice 10.

[0082] The radiator 110 in the analysis device 100 emits linearly polarized excitation light EL in the -X direction, such as Figure 8 As shown. Radiator 110 includes a light source 112, an excitation light filter 114, a polarization filter 116, and optical components (not shown), such as a condenser lens, etc. Figure 7 and Figure 8 As shown. Light source 112 emits light including excitation light EL in the -X direction. For example, light source 112 is configured with LED elements. Excitation light filter 114 removes light other than excitation light EL from the light emitted from light source 112. For example, excitation light filter 114 is a bandpass filter. Polarization filter 116 converts the excitation light EL passing through excitation light filter 114 into linearly polarized light.

[0083] The dichroic mirror 120 in the analysis device 100 reflects the linearly polarized excitation light EL emitted from the radiator 110 in a direction (+Z direction) toward the location where the micro-device 10 is placed, such as... Figure 8 As shown. In addition, the dichroic mirror 120 transmits the fluorescence FL emitted from the microdevice 10.

[0084] In the analytical apparatus 100, the microdevice 10 is placed on the +Z side of the dichroic mirror 120, with the first substrate 12 facing the Z direction. Linearly polarized excitation light EL reflected by the dichroic mirror 120 enters the microchannel 18 from the first substrate 12 in the microdevice 10, as... Figure 2 and Figure 3 As shown. Furthermore, the microdevice 10 emits fluorescence FL in the Z direction.

[0085] The objective lens 130 in the analysis device 100 is placed between the dichroic mirror 120 and the microdevice 10, such as Figure 8 As shown. Objective 130 converges excitation light (EL) and fluorescence (FL).

[0086] The detector 140 in the analysis device 100 is placed on the -Z side of the dichroic mirror 120, such as Figure 8 As shown. Detector 140 detects the fluorescent FL emitted from microdevice 10. (As illustrated...) Figure 7 and Figure 8 As shown, detector 140 includes an absorption filter 142, a polarization adjustment element 144, an imaging element 146, and optical components (not shown), such as an imaging lens. The absorption filter 142 separates the emitted fluorescence FL from the microdevice 10 from scattered light, leaked light, etc., and transmits the emitted fluorescence FL. For example, the absorption filter 142 is a bandpass filter. The polarization adjustment element 144 adjusts the polarization direction of the fluorescence FL transmitted through the absorption filter 142. The polarization adjustment element 144 adjusts the polarization direction of the fluorescence FL to a direction parallel to the polarization direction of the excitation light EL emitted from the radiator 110 and perpendicular to the polarization direction of the excitation light EL emitted from the radiator 110. For example, the polarization adjustment element 144 is a liquid crystal element. The imaging element 146 detects the fluorescence FL emitted from the polarization adjustment element 144 as an image. For example, the imaging element 146 is a charge-coupled device (CCD) image sensor.

[0087] The controller 150 in the analysis device 100 controls the radiator 110 and the detector 140. Further, the controller 150 determines the polarization degree P of the fluorescent FL emitted from the microdevice 10 based on an image of the fluorescent FL detected by the imaging element 146. Additionally, the controller 150 determines the concentration of the target substance Ag1 based on the polarization degree P and a previously generated calibration curve. The controller 150 includes a central processing unit (CPU) 152 that performs various types of processing, a read-only memory (ROM) 154 that stores programs and data, a random access memory (RAM) 156 that stores data, and an input / output interface 158 that inputs and outputs signals between the components. The functionality of the controller 150 is provided by the CPU 152, which executes programs stored in the ROM 154. The input / output interface 158 inputs and outputs signals between the CPU 152, the radiator 110, and the detector 140.

[0088] The operation of the analytical apparatus 100 and the detection of the target substance Ag1 will be described.

[0089] First, the target solution is introduced into the microchannel 18 of the microdevice 10, and then the microdevice 10 is placed at a predetermined position in the analytical device 100. Next, after the competitive reaction between the target substance Ag1 and the fluorescently labeled derivative AgF1 in the microchannel 18 reaches equilibrium, linearly polarized excitation light EL is emitted from the radiator 110 in the analytical device 100, and the radiation region S in the microdevice 10 is irradiated by the linearly polarized excitation light EL through the dichroic mirror 120 and the objective lens 130, as shown... Figure 8 As shown.

[0090] Linearly polarized excitation light EL enters the microdevice 10 from the first substrate 12, such as Figure 3 As shown. In this case, the first light blocker 16 blocks the excitation light EL entering the bound fluorescently labeled derivative AgF1, and thus suppresses the fluorescence of the bound fluorescently labeled derivative AgF1, while the fluorescence FL radiated by the free fluorescently labeled derivative AgF1 in the microchannel 18 is emitted from the microdevice 10 in the -Z direction. Since the competitive reaction has reached equilibrium, the amount of free fluorescently labeled derivative AgF1 in the microchannel 18 is related to the concentration of the target substance Ag1 being measured.

[0091] Fluorescent light emitted from the microdevice 10 passes through the objective lens 130 and the dichroic mirror 120 and enters the detector 140, such as Figure 8As shown. Detector 140 adjusts the polarization direction of the fluorescence FL via polarization adjustment element 144. Then, detector 140 acquires images of the fluorescence FL with a polarization direction parallel to the excitation light EL and images of the fluorescence FL with a polarization direction perpendicular to the excitation light EL via imaging element 146.

[0092] The controller 150 determines the degree of polarization of the fluorescent FL based on the acquired image. Ih represents the intensity of the fluorescent FL with a polarization direction parallel to the excitation light EL, and Iv represents the intensity of the fluorescent FL with a polarization direction perpendicular to the excitation light EL. The degree of polarization of the fluorescent FL is expressed as P = (Ih - Iv) / (Ih + Iv). Furthermore, the controller 150 determines the concentration of the target substance Ag1 based on the degree of polarization P and a previously generated calibration curve. Therefore, the concentration of the target substance Ag1 can be obtained.

[0093] In the analytical apparatus 100, the fluorescence of the bound fluorescently labeled derivative AgF1 is suppressed, and the fluorescence FL emitted by the free fluorescently labeled derivative AgF1 is detected. Therefore, the analytical apparatus 100 is able to measure the concentration of the target substance Ag1 with high measurement sensitivity.

[0094] As described above, in the microdevice 10, the first light blocker 16 blocks the excitation light EL entering the bound fluorescently labeled derivative AgF1, and therefore, when the target solution is introduced into the microchannel 18, the fluorescence of the bound fluorescently labeled derivative AgF1 is suppressed, and the fluorescence FL radiated by the free fluorescently labeled derivative AgF1 is emitted. Therefore, the microdevice 10 can increase the measurement sensitivity to the target substance Ag1. The microdevice 10 can suppress the fluorescence of the bound fluorescently labeled derivative AgF1, and therefore can detect the target substance Ag1 with a higher molecular weight. Furthermore, the microdevice 10 can detect the target substance Ag1 without adding the fluorescently labeled derivative AgF1 to the target solution, and therefore can easily detect the target substance Ag1.

[0095] The analytical device 100 suppresses the fluorescence of the bound fluorescently labeled derivative AgF1 and detects the fluorescence FL emitted by the free fluorescently labeled derivative AgF1. Therefore, the analytical device 100 can detect the target substance Ag1 with high measurement sensitivity. Furthermore, the analytical device 100 can easily detect the target substance Ag1. Additionally, the analytical device 100 determines the concentration of the target substance Ag1 based on the degree of polarization P, and thus can suppress the effects of variations in the intensity of light emitted by the excitation light EL and the light emitted by the light source 112, and can determine the concentration of the target substance Ag1 with high accuracy.

[0096] Example 2

[0097] Although the first light blocker 16 forms the microchannel 18 in Embodiment 1, another component can form the microchannel 18. Furthermore, the second light blocker 26 that blocks the excitation light EL can be disposed on another component.

[0098] Reference Figures 9 to 11 The micro device 10 according to this embodiment is described. For example... Figure 9 and Figure 10 As shown, the microdevice 10 according to this embodiment includes a first substrate 12, a second substrate 14, three microchannels 18, a partition wall 22, and a second light blocker 26. Additionally, the microdevice 10 according to this embodiment includes an antibody Ab1 and a fluorescently labeled derivative AgF1, such as... Figure 11 As shown. In this embodiment, the partition wall 22 forms a microchannel 18. Furthermore, a second light blocker 26 is disposed on the first substrate 12. The second light blocker 26 blocks the excitation light EL from the bound fluorescently labeled derivative AgF1.

[0099] According to this embodiment, the second light blocker 26 is disposed on the second main plane 12b of the first substrate 12, such as... Figure 10 As shown. According to this embodiment, the first substrate 12 and the second substrate 14 sandwich the partition wall 22 in the middle. The remaining configuration of the first substrate 12 according to this embodiment is similar to the configuration according to embodiment 1.

[0100] According to this embodiment, the second substrate 14 and the first substrate 12 sandwich the partition wall 22 between them. Furthermore, according to this embodiment, the second substrate 14 and the partition wall 22 are integrally formed. The remaining configuration of the second substrate 14 according to this embodiment is similar to the configuration according to Embodiment 1.

[0101] The partition wall 22 is sandwiched between the first substrate 12 and the second substrate 14, forming a microchannel 18. The partition wall 22 is formed of a material with low autofluorescence. Further, the partition wall 22 is preferably formed of a material that absorbs light such as excitation light EL and fluorescence FL. In this embodiment, the partition wall 22 is integrally formed of polydimethylsiloxane containing carbon black with the second substrate 14.

[0102] When viewed in a cross-section (YZ plane) along the width direction of the microchannel 18, the side portion 22a of the partition wall 22 forms the sidewall surface 18a of the microchannel 18. In this embodiment, the two side portions 22a forming the two sidewall surfaces 18a of the microchannel 18 are perpendicular to the second main plane 12b of the first substrate 12 and the first main plane 14a of the second substrate 14. Figure 11As shown, antibody Ab1 is immobilized to the side portion 22a of septum 22 (i.e., the sidewall surface 18a of microchannel 18) in a state that specifically binds to the fluorescently labeled derivative AgF1.

[0103] In the microchannel 18 according to this embodiment, the sidewall surface 18a is formed by the side portion 22a of the partition wall 22. Furthermore, the microchannel 18 according to this embodiment has a rectangular cross-section in the width direction (Y direction). The remaining configuration of the microchannel 18 according to this embodiment is similar to the configuration according to Embodiment 1.

[0104] According to this embodiment, antibody Ab1 is immobilized on the sidewall surface 18a of the microchannel 18 (i.e., the side portion 22a of the septum 22). Further, according to this embodiment, the fluorescently labeled derivative AgF1 specifically binds to antibody Ab1 via an antigen-antibody reaction. The configuration of antibody Ab1 and fluorescently labeled derivative AgF1 according to this embodiment is similar to the configuration according to Example 1.

[0105] The second light blocker 26 according to this embodiment is disposed on the second main plane 12b of the first substrate 12 and blocks the excitation light EL of the bound fluorescent label derivative AgF1. For example, the second light blocker 26 according to this embodiment is formed of a resin containing carbon black.

[0106] In the microdevice 10 according to this embodiment, similar to the microdevice 10 according to Embodiment 1, a second light blocker 26 disposed on the first substrate 12 blocks the excitation light EL entering the bound fluorescently labeled derivative AgF1. Therefore, when the target solution is introduced into the microchannel 18, the fluorescence of the bound fluorescently labeled derivative AgF1 is suppressed, and fluorescence radiated by the free fluorescently labeled derivative AgF1 is emitted. Thus, similar to the microdevice 10 according to Embodiment 1, the microdevice 10 according to this embodiment can improve the measurement sensitivity to the target substance Ag1. Furthermore, the microdevice 10 according to this embodiment can detect target substances Ag1 with higher molecular weights. Additionally, similar to the microdevice 10 according to Embodiment 1, the microdevice 10 according to this embodiment can easily detect the target substance Ag1.

[0107] Example 3

[0108] Although in Examples 1 and 2 one antibody Ab1 was immobilized into microchannel 18, multiple antibodies Ab1, Ab2 and Ab3 can be immobilized into one microchannel 18.

[0109] Reference Figures 12 to 19 The micro device 10 according to this embodiment is described. For example... Figure 12As shown, the microdevice 10 according to this embodiment includes a microchannel 18. In this embodiment, a first substrate 12, a second substrate 14, third light blockers 161 to 166, and a fourth light blocker 170 form a microchannel 18. The microchannel 18 in the radiation region S includes three segments R1, R2, and R3 along the length direction (-X direction). Furthermore, as... Figure 13 As shown, the sidewall surface 18a of the microchannel 18 in the radiation region S is divided into segments R1, R2, and R3 corresponding to the segments R1, R2, and R3 of the microchannel 18 along the length direction of the microchannel 18. Additionally, as... Figures 14 to 16 As shown, antibodies Ab1, Ab2, and Ab3 are immobilized in the R1, R2, and R3 segments of the sidewall surface 18a, respectively, and specifically bind to the fluorescently labeled derivatives AgF1, AgF2, and AgF3, respectively. The remaining configuration is similar to that according to Example 1. The configuration of the microdevice 10 according to this embodiment will be described in detail below.

[0110] In this embodiment, the third light blockers 161 to 166 and the fourth light blocker 170 are sandwiched between the first substrate 12 and the second substrate 14, forming a microchannel 18. The third light blockers 161 to 166 form the microchannel 18 in the radiation region S, such as... Figure 12 As shown. Each of the third light blockers 161 to 166 has a tetragonal prism shape, and the lower surface of the tetragonal prism shape (the cross-section of the microchannel 18 in the width direction) is a rectangular trapezoid, as shown. Figure 13 As shown. Third light blockers 161, 163, and 165 are arranged in the X direction, and the corresponding inclined sides 161a, 163a, and 165a of the third light blockers 161, 163, and 165 form one sidewall surface 18a of the microchannel 18. Third light blockers 162, 164, and 166 are arranged in the X direction, and the corresponding inclined sides 162a, 164a, and 166a of the third light blockers 162, 164, and 166 form the other sidewall surface 18a of the microchannel 18.

[0111] Moreover, such as Figures 13 to 16 As shown, the third light blocker 161 and the third light blocker 162 face each other, and the side portion 161a of the third light blocker 161 and the side portion 162a of the third light blocker 162 form the dividing portion R1 of the two sidewall surfaces 18a. Similar to the third light blockers 161 and 162, the side portion 163a of the third light blocker 163 and the side portion 164a of the third light blocker 164 form the dividing portion R2 of the sidewall surface 18a, and the side portion 165a of the third light blocker 165 and the side portion 166a of the third light blocker 166 form the dividing portion R3 of the sidewall surface 18a.

[0112] In addition to the microchannel 18 formed by the third light blockers 161 to 166, a fourth light blocker 170 forms the microchannel 18. The fourth light blocker 170 includes a recess 172 into which the third light blockers 161 to 166 are fitted, such as... Figure 17 As shown. In this embodiment, the fourth light blocker 170 is integrally formed with the second substrate 14.

[0113] In this embodiment, antibody Ab1 is immobilized on the segment R1 of the sidewall surface 18a in a state bound to a fluorescently labeled derivative AgF1 obtained by fluorescently labeling the target substance Ag1. The target substance Ag1 and the fluorescently labeled derivative AgF1 specifically bind to antibody Ab1, but not to either antibody Ab2 or antibody Ab3. Furthermore, antibody Ab2 is immobilized on the segment R2 of the sidewall surface 18a in a state bound to a fluorescently labeled derivative AgF2 obtained by fluorescently labeling the target substance Ag2. The target substance Ag2 and the fluorescently labeled derivative AgF2 specifically bind to antibody Ab2, but not to either antibody Ab1 or antibody Ab3. Additionally, antibody Ab3 is immobilized on the segment R3 of the sidewall surface 18a in a state bound to a fluorescently labeled derivative AgF3 obtained by fluorescently labeling the target substance Ag3. The target substance Ag3 and the fluorescently labeled derivative AgF3 were measured to specifically bind to antibody Ab3, but not to either antibody Ab1 or antibody Ab2.

[0114] In this embodiment, similar to the side 16a of the first light blocker 16 according to Embodiment 1, the sides 161a to 166a (sidewall surfaces 18a of the microchannel 18) of the third light blockers 161 to 166 are inclined in a direction that narrows the width of the microchannel 18 toward the first substrate 12. Therefore, similar to the first light blocker 16 according to Embodiment 1, the third light blockers 161 to 166 block the excitation light EL entering the bound fluorescently labeled derivatives AgF1, AgF2, and AgF3, and similar to the bound fluorescently labeled derivative AgF1 according to Embodiment 1, the bound fluorescently labeled derivatives AgF1, AgF2, and AgF3 do not radiate fluorescence even when the microdevice 10 is irradiated by the excitation light EL.

[0115] Next, the operation of the microdevice 10 according to this embodiment will be described.

[0116] When a measurement target solution containing the measurement target substances Ag1, Ag2, and Ag3 is introduced into the microchannel 18 of the microdevice 10, the measurement target substance Ag1 specifically binds to antibody Ab1 in the segmentation R1 on the sidewall surface 18a through an antigen-antibody reaction competing with the fluorescently labeled derivative AgF1. Then, free fluorescently labeled derivative AgF1 is generated in the segmentation R1 of the microchannel 18. Further, similar to the free fluorescently labeled derivative AgF1, free fluorescently labeled derivative AgF2 is generated in the segmentation R2 of the microchannel 18, and free fluorescently labeled derivative AgF3 is generated in the segmentation R3 of the microchannel 18.

[0117] When the competitive reaction reaches equilibrium, free fluorescently labeled derivatives AgF1, AgF2, and AgF3 are generated in amounts determined according to the concentrations of the target substances Ag1, Ag2, and Ag3. When excitation light EL enters the first substrate 12 after the competitive reaction has reached equilibrium, third photoblocks 161 to 166 block the excitation light EL from the bound fluorescently labeled derivatives AgF1, AgF2, and AgF3, and thus measure the fluorescence emitted by the free fluorescently labeled derivatives AgF1, AgF2, and AgF3. Therefore, the concentrations of the target substances Ag1, Ag2, and Ag3 can be determined based on the polarization degree P of the divisions R1, R2, and R3 of the microchannel 18. Furthermore, similar to the microdevice 10 according to Embodiment 1, the microdevice 10 according to this embodiment can improve the measurement sensitivity of the target substances Ag1, Ag2, and Ag3.

[0118] Next, we will refer to Figure 18 and Figure 19 A method for manufacturing the microdevice 10 according to this embodiment is described. Figure 18 This is a flowchart illustrating a method for manufacturing the microdevice 10 according to this embodiment. For ease of understanding, antibodies Ab1, Ab2, and Ab3 can be collectively referred to as antibodies Ab in the following text, and fluorescently labeled derivatives AgF1, AgF2, and AgF3 can be collectively referred to as fluorescently labeled derivatives AgF.

[0119] The manufacturing method of the microdevice 10 according to this embodiment includes the following steps: preparing third light blockers 161 to 166 and components that are integrally formed as second substrate 14 and fourth light blocker 170 (step S110); immobilizing antibody Ab to the side portions 161a to 166a of third light blockers 161 to 166 and specifically binding fluorescently labeled derivative AgF to the immobilized antibody Ab (step S120); bonding third light blockers 161 to 166 to fourth light blocker 170 (step S130); and bonding third light blockers 161 to 166 and fourth light blocker 170 to first substrate 12 (step S140).

[0120] In step S110, third light blockers 161 to 166 are prepared. Third light blockers 161 to 166 are cut from a polydimethylsiloxane resin block containing carbon black. Further, a component is prepared as an integrally formed part of the second substrate 14 and the fourth light blocker 170. Similar to step S10 according to Embodiment 1, the component as an integrally formed part of the second substrate 14 and the fourth light blocker 170 is formed using a mold. Additionally, similar to step S20 according to Embodiment 1, an inlet 19a and an outlet 19b are formed using a fixture.

[0121] Step S120 will be described using third light blockers 161 and 162 as examples. First, by sandwiching third light blockers 161 and 162 between two glass substrates 301 and 302, a channel 303 is formed with the side portions 161a of third light blocker 161 and 162a of third light blocker 162 as sidewall surfaces, as shown. Figure 19 As shown. Next, a solution containing antibody Ab1 is introduced into the formed channel 303, and antibody Ab1 is immobilized to side portions 161a of the third light blocker 161 and 162a of the third light blocker 162. Next, a solution containing a fluorescently labeled derivative AgF1 is introduced into the channel 303, and the fluorescently labeled derivative AgF1 binds to antibody Ab1 via an antigen-antibody reaction. Therefore, antibody Ab1 can be immobilized to side portions 161a and 162a, and the fluorescently labeled derivative AgF1 can specifically bind to antibody Ab1. Similar to the third light blockers 161 and 162, antibody Ab2 is immobilized relative to the third light blockers 163 and 164, and the fluorescently labeled derivative AgF2 specifically binds to antibody Ab2. Furthermore, similar to the third light blockers 161 and 162, relative to the third light blockers 165 and 166, antibody Ab3 is immobilized, and the fluorescently labeled derivative AgF3 specifically binds to antibody Ab3.

[0122] In step S130, firstly, the third light blockers 161 to 166 are placed in the groove 172 of the fourth light blocker 170. Next, the third light blockers 161 to 166 and the fourth light blocker 170 are joined to the fourth light blocker 170 by pressing them together. Thus, the sidewall surface 18a of the microchannel 18, including the divisions R1, R2, and R3, is formed.

[0123] In step S140, firstly, the first substrate 12 is placed on the third light blockers 161 to 166 and the fourth light blocker 170. Then, by pressing the first substrate 12 against the third light blockers 161 to 166 and the fourth light blocker 170, the third light blockers 161 to 166 and the fourth light blocker 170 are bonded to the first substrate 12. Thus, a microdevice 10 according to this embodiment can be produced.

[0124] As described above, in this embodiment, the third light blockers 161 to 166 block the excitation light EL entering the bound fluorescently labeled derivatives AgF1, AgF2, and AgF3, and therefore the fluorescence of the bound fluorescently labeled derivatives AgF1, AgF2, and AgF3 is suppressed when the target solution is introduced into the microchannel 18. Therefore, similar to the microdevice 10 according to Embodiment 1, the microdevice 10 can increase the measurement sensitivity for the target substances Ag1, Ag2, and Ag3. Further, the sidewall surface 18a of the microchannel 18 in the radiation region S is divided into multiple segments R1, R2, and R3 along the length of the microchannel 18, and multiple antibodies Ab1, Ab2, and Ab3 are fixed for each segment; and therefore, multiple target substances Ag1, Ag2, and Ag3 can be detected. Similar to the microdevice 10 according to Embodiment 1, the microdevice 10 according to this embodiment can easily detect the target substances Ag1, Ag2, and Ag3. In addition, the micro device 10 according to this embodiment can detect the target substances Ag1, Ag2 and Ag3 with high molecular weight.

[0125] Example 4

[0126] Although in Examples 1 to 3, the antibody Ab was immobilized on the sidewall surface 18a in a state of specific binding to the fluorescently labeled derivative AgF, the antibody Ab may not bind to the fluorescently labeled derivative AgF.

[0127] In the microdevice 10 according to this embodiment, the antibody Ab1 immobilized on the sidewall surface 18a does not specifically bind to the fluorescently labeled derivative AgF1, such as Figure 20 As shown. The remaining configuration of the microdevice 10 according to this embodiment is similar to the configuration of the microdevice 10 according to embodiment 1.

[0128] In the microdevice 10 according to this embodiment, a measurement target solution (hereinafter referred to as the measurement target solution) comprising a measurement target substance Ag1 at a predetermined concentration and a fluorescently labeled derivative AgF1 is introduced into a microchannel 18. When the measurement target solution is introduced into the microchannel 18, the measurement target substance Ag1 and the fluorescently labeled derivative AgF1 competitively induce an antigen-antibody reaction against antibody Ab1. Similar to Example 1, when the competitive reaction between the measurement target substance Ag1 and the fluorescently labeled derivative AgF1 against antibody Ab1 reaches equilibrium, free fluorescently labeled derivative AgF1 in an amount determined according to the concentration of the measurement target substance Ag1 is generated in the microchannel 18.

[0129] In the microdevice 10 according to this embodiment, similar to the microdevice 10 according to Embodiment 1, a first light blocker 16 blocks the excitation light EL entering the bound fluorescently labeled derivative AgF1. Therefore, the microdevice 10 according to this embodiment can suppress the fluorescence of the bound fluorescently labeled derivative AgF1 and improve the measurement sensitivity to the target substance Ag1. The microdevice 10 according to this embodiment can detect the target substance Ag1 with a high molecular weight.

[0130] Modified example

[0131] Although embodiments have been described above, various modifications may be made to this disclosure without departing from its spirit.

[0132] Although the second substrate 14 and the first light blocker 16 are integrally formed in Embodiment 1, the second substrate 14 and the first light blocker 16 can be formed separately. Furthermore, the second substrate 14 can transmit excitation light EL.

[0133] The second substrate 14, the first light blocker 16, the partition wall 22, the third light blockers 161 to 166, and the fourth light blocker 170 are formed of polydimethylsiloxane containing carbon black, but can also be formed of another material. For example, polydimethylsiloxane can contain iron oxide instead of carbon black. Furthermore, the first light blocker 16, the third light blockers 161 to 166, etc., can be formed of polymethyl methacrylate containing iron oxide.

[0134] In the microdevice 10 according to Example 1, antibody Ab1 is immobilized to both sidewall surfaces 18a of the microchannel 18 in a state specifically bound to the fluorescently labeled derivative AgF1. According to Example 1, antibody Ab1 must only be immobilized to at least one of the sidewall surfaces 18a of the microchannel 18 in a state specifically bound to the fluorescently labeled derivative AgF1. When antibody Ab1 is immobilized to one of the sidewall surfaces 18a, only the sidewall surface 18a to which antibody Ab1 is immobilized needs to be inclined in the direction that narrows the width of the microchannel 18 toward the first substrate 12.

[0135] Similar to Example 3, for each segment, each of the plurality of antibody Abs can be fixed to the sidewall surface 18a of the microchannel 18 according to Example 1.

[0136] In Embodiment 1, the entire sidewall surface 18a of the microchannel 18 is inclined in a direction that narrows the width of the microchannel 18 toward the first substrate 12. A portion of the sidewall surface 18a according to Embodiment 1 may be inclined in the direction that narrows the width of the microchannel 18. For example, the end of the sidewall surface 18a on the first substrate 12 side may be bent toward the interior of the microchannel 18, such as... Figure 21 As shown. In this case, for example, the second substrate 14 and the first light blocker 16 are used by Figure 22 The mold 310 shown is formed. For example, the mold 310 is produced by etching and oxidizing the silicon layer 312 in the silicon on insulator (SOI) substrate and then wet etching the oxide film and silicon oxide film layer 314 formed in the SOI substrate.

[0137] The analytical device 100 may include one of the microdevices 10 according to Embodiments 2 to 4 instead of the microdevice 10 according to Embodiment 1. Further, the analytical device 100 can determine the concentration of the target substance Ag1 based on the intensity of the fluorescence FL. For example, the analytical device 100 can detect the intensity Iv of the fluorescence FL having a polarization direction perpendicular to the excitation light EL using a detector, and determine the concentration of the target substance Ag1 based on Iv. Therefore, the S / N ratio of the fluorescence is improved, and the measurement sensitivity is enhanced. Additionally, the imaging element 146 in the detector 140 is not limited to a CCD image sensor. For example, the imaging element 146 in the detector 140 may be a complementary metal oxide semiconductor (CMOS) image sensor.

[0138] The analysis device 100 may include a promoter 180, such as Figure 23As shown. Promoter 180 facilitates a competitive reaction between the target substance Ag and the fluorescently labeled derivative AgF against the antibody Ab. Examples of promoter 180 include a heater for heating the microdevice 10 and an ultrasonic device that uses ultrasound to irradiate the microdevice 10.

[0139] Although the second substrate 14 and the partition wall 22 are integrally formed in Embodiment 2, the second substrate 14 and the partition wall 22 can be formed separately. Furthermore, the second substrate 14 and the partition wall 22 can transmit excitation light EL.

[0140] In Embodiment 2, when viewed in a cross-section (YZ plane) of the microchannel 18 in the width direction, the side portion 22a of the partition wall 22 does not need to be perpendicular to the second main plane 12b of the first substrate 12 or the first main plane 14a of the second substrate 14. When viewed in a cross-section (YZ plane) of the microchannel 18 in the width direction, the microchannel 18 may have a tapered shape.

[0141] Similar to Example 3, for each segment, each of the plurality of antibody Abs can be immobilized to the sidewall surface 18a of the microchannel 18 according to Example 2. Additionally, similar to Example 4, antibody Abs that do not specifically bind to the fluorescently labeled derivative AgF can be immobilized to the sidewall surface 18a of the microchannel 18 according to Example 2.

[0142] Although in Embodiment 2 the second light blocker 26 is disposed on the second main plane 12b of the first substrate 12, a second light blocker 26 can be disposed on the partition wall 22 to block the excitation light EL entering the bound fluorescently labeled derivative AgF1. For example, the second light blocker 26 can be disposed at the end of the partition wall 22 on the first substrate 12 side, such as... Figure 24 As shown. In this case, the partition wall 22 and the second light blocker 26 can be integrally formed.

[0143] The microdevice 10 according to embodiment 3 may include a plurality of microchannels 18. Furthermore, the number of segments of the microchannels 18 is not limited to three, and only one is required.

[0144] In each of the microdevices 10 according to Examples 1 to 3, an antibody Ab specifically binding to the fluorescently labeled derivative AgF can be immobilized on at least one of the upper wall surface 18b and the lower wall surface 18c of the microchannel 18. The surface area of ​​each of the upper wall surface 18b and the lower wall surface 18c is smaller than that of the sidewall surface 18a, and therefore the detection of the target substance Ag is hardly affected even when the antibody Ab specifically binding to the fluorescently labeled derivative AgF is immobilized on at least one of the upper wall surface 18b and the lower wall surface 18c of the microchannel 18. The antibody Ab can also be immobilized on at least one of the upper wall surface 18b and the lower wall surface 18c of the microchannel 18 in the microdevice 10 according to Example 4.

[0145] In the manufacture of the microdevice 10, for example, by immobilizing antibody Abs to the sidewall surface 18a, the upper wall surface 18b, and the lower wall surface 18c and then removing the antibody Abs immobilized to the upper wall surface 18b and the lower wall surface 18c by using laser radiation, antibody Abs or antibody Abs specifically bound to fluorescently labeled derivative AgF can be immobilized only to the sidewall surface 18a.

[0146] For illustrative purposes, some exemplary embodiments have been described above. Although specific embodiments have been presented in the foregoing discussion, those skilled in the art will recognize that changes in form and detail may be made without departing from the broader spirit and scope of the invention. Therefore, the specification and drawings are to be viewed in an illustrative rather than restrictive sense. Consequently, this detailed description should not be construed in a restrictive sense, and the scope of the invention is defined only by the included claims and the full scope of their equivalents.

Claims

1. A miniature device, comprising: A microchannel, into which a target solution containing the target substance is introduced; Antibodies are immobilized on at least one sidewall surface of the microchannel and specifically bind to the target substance being measured. A fluorescently labeled derivative, which specifically binds to the antibody and is obtained by fluorescently labeling the target substance being measured; as well as A light blocker blocks the excitation light that excites fluorescence emitted by the fluorescently labeled derivative, wherein... The target substance and the fluorescently labeled derivative bind specifically to the antibody in a competitive manner. The antibody is immobilized on the sidewall surface of the microchannel in a state that specifically binds to the fluorescently labeled derivative, and The light blocker prevents the excitation light from entering the sidewall surface, thereby preventing the excitation light from entering the fluorescently labeled derivative that specifically binds to the antibody.

2. The microdevice according to claim 1, wherein The antibody is a variety of antibodies. The fluorescently labeled derivatives are a variety of fluorescently labeled derivatives. The target solution for measurement includes multiple target substances, and Each of the plurality of antibodies specifically binds to one of the plurality of measurement target substances and one of the plurality of fluorescently labeled derivatives, and is immobilized for each segment of the sidewall surface along the length of the microchannel.

3. A miniature device, comprising: A microchannel is introduced into the microchannel, comprising a target substance for measurement and a fluorescently labeled derivative obtained by fluorescently labeling the target substance for measurement. Antibodies are immobilized on at least one sidewall surface of the microchannel and specifically bind to the target substance being measured and the fluorescently labeled derivative. as well as A light blocker blocks the excitation light that excites fluorescence emitted by the fluorescently labeled derivative, wherein... The target substance and the fluorescently labeled derivative bind specifically to the antibody in a competitive manner, and, When the target solution is introduced into the microchannel, the light blocker blocks the excitation light from entering the sidewall surface, thereby preventing the excitation light from entering the fluorescently labeled derivative that specifically binds to the antibody.

4. The microdevice according to claim 3, wherein The antibody is a variety of antibodies. The target solution contains multiple target substances and multiple fluorescently labeled derivatives, and Each of the plurality of antibodies specifically binds to one of the plurality of measurement target substances and one of the plurality of fluorescently labeled derivatives, and is immobilized for each segment of the sidewall surface along the length of the microchannel.

5. The microdevice according to any one of claims 1 to 4, further comprising: The first substrate into which the excitation light enters; as well as The second substrate facing the first substrate, wherein The light blocker forms the microchannel by being sandwiched between the first substrate and the second substrate. The side portion of the light blocker forms the sidewall surface of the microchannel, and The sidewall surface of the microchannel to which the antibody is attached is inclined in a direction that narrows the width of the microchannel toward the first substrate.

6. The microdevice of claim 5, wherein the cross-section of the microchannel in the width direction has a tapered shape that narrows toward the first substrate.

7. The microdevice according to any one of claims 1 to 4, further comprising: The excitation light enters the first substrate; The second substrate facing the first substrate, and The partition walls of the microchannel are formed by being sandwiched between the first substrate and the second substrate. The side portion of the partition wall forms the sidewall surface of the microchannel.

8. The microdevice according to claim 7, wherein the light blocker is disposed on the first substrate.

9. The microdevice of claim 7, wherein the light blocker is disposed at the end of the partition wall on the first substrate side.

10. The microdevice according to claim 1 or 3, wherein the light blocker is formed of a material that absorbs the excitation light and the fluorescence radiated by the fluorescently labeled derivative.

11. The microdevice of claim 10, wherein the light blocker is formed of polydimethylsiloxane containing carbon black.

12. An analytical apparatus, comprising: The microdevice according to any one of claims 1 to 11; A radiator that uses the excitation light transmitted through a light blocker to radiate the microchannels of the microdevice; as well as The detector detects the fluorescence.

13. The analytical apparatus according to claim 12, wherein The excitation light is linearly polarized light with a polarization direction in a predetermined direction, and The fluorescence is linearly polarized light having a polarization direction orthogonal to the predetermined direction.

14. The analytical apparatus according to claim 12 or 13, further comprising: The promoter facilitates a competitive response between the target substance and the fluorescently labeled derivative to the antibody.

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