JP3528800B2 - Optical biomolecule detection device and optical biomolecule detection method - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an immunodiagnostic sensor utilizing the optical characteristics of precious metal fine particles, a DNA chip, a protein chip, and a measuring apparatus using the same.

[0002]

2. Description of the Related Art As a conventional sensor of this type, there is a sensor utilizing a surface plasmon resonance method. The surface plasmon is a compressional wave of free electrons propagating at the interface between the metal thin film and the dielectric, and is greatly affected by the dielectric constant at the interface, and is therefore used in the detection principle of immunosensors, gas sensors and the like. FIG. 2 shows a specific structural example of a measuring device to which this sensor is applied. A thin film 22 of free electron metal such as gold or silver having a thickness of about 50 nm is formed on the surface of a transparent high refractive index carrier 21 such as a prism, and a molecular recognition layer 23 is formed on the thin film 22. In order to excite the surface plasmons of the thin film 22, p-polarized monochromatic parallel light 24 is emitted from the light source 25 from the prism side. Excitation of the surface plasmons can be confirmed by detecting the specularly reflected light 27 with the detector 28 while changing the incident angle 26 under the condition of total reflection. That is, at the resonance incident angle 29 at which the surface plasmons are excited, the energy of the incident light is consumed for the surface plasmon excitation, so that the intensity 30 of the reflected light is extremely reduced. When the target biomolecule is captured by the molecule recognition layer 23, the intensity 32 of the reflected light is extremely reduced at the resonance incident angle 31. Since the resonance angle sensitively depends on the dielectric constant in a region within several hundred nm from the interface, knowing the resonance angle enables the dielectric constant of the molecule existing in the molecule recognition layer 23 existing on the metal surface to be specified. It can be used as a sensor. For example, a structure for recognizing a specific molecule and forming a molecular bond is formed on the surface of the thin film 22, and when the specific molecule is bonded, the dielectric constant changes. Therefore, the reflected light at the reflection angle corresponding to the molecule is formed. It is possible to immediately know that a specific molecule has been captured by the molecular recognition layer 23 by monitoring.

[0003]

In the measuring device using the surface plasmon sensor of the above-mentioned prior art, in order to measure the resonance incident angle, the positional relationship between the light source of the irradiation light, the metal thin film, and the photodetector is accurately maintained. And needs to be driven. In order to improve the measurement accuracy, it is desirable to increase the distance between the metal thin film and the photodetector, but this is incompatible with miniaturization and arraying for the purpose of detecting multiple samples. Also,
Since the surface plasmon resonance method is sensitive to temperature, it is necessary to control the temperature of the entire apparatus or correct the temperature, which is also unsuitable for downsizing.

[0004]

In order to solve the above problems, an optical phenomenon of noble metal fine particles prepared by the method recently invented by the inventors is utilized. The noble metal fine particles that change color when biomolecules are adsorbed on the surface are formed on the surface of the cartridge, and after the reaction, the cartridge is inserted into an optical measurement device and the optical characteristics are measured, whereby the biomolecules can be easily and quickly bound. A biomolecule detecting device comprising an insertion port for inserting a cartridge having a surface on which a single layer of noble metal-coated dielectric fine particles is formed and an optical measuring device for measuring optical characteristics of the surface of the cartridge is used. A cartridge is used in which the surface of the cartridge is divided into a plurality of regions, and each region is composed of fine particles having different optical characteristics or fine particles modified with different biomolecules. The biomolecule detecting device is characterized in that the surface of the cartridge has a plurality of regions, and has means for scanning the detection probe light on the plurality of regions. The surface of the cartridge has a plurality of areas, and a means for moving the cartridge with respect to the detection probe light whose position is fixed is used. The biomolecule detecting device is characterized in that the surface of the cartridge has a plurality of regions, and the biomolecule detecting device has means for irradiating the plurality of regions at the same time and measuring optical characteristics. The insertion port for inserting the cartridge in which the biomolecular reaction has been completed, and the optical property data stored in advance in the personal computer connected to the optical measuring device with the detected optical property, or the reference area formed on the surface of the cartridge. A biomolecule detecting device having a means for comparing with an optical characteristic is used. When detecting a biomolecule with the biomolecule detection device, a biomolecule sample is supplied from the outside to the surface of the inserted cartridge, and the optical characteristic of the surface of the cartridge is measured, and the biomolecule detection device is used. . A biomolecule detecting device having an insertion opening for inserting the cartridge in combination with an adapter is used. A biomolecule detecting device is provided which has a means for optically, magnetically or mechanically marking after using the cartridge.
According to a biomolecule detection method characterized by detecting a biomolecule, which comprises a step of carrying out a biomolecule reaction on the surface of the cartridge, a step of inserting the cartridge into an apparatus, and a step of comparison. Further description will be made with reference to the drawings. FIG. 1 shows the device configuration. By inserting a rod-shaped cartridge 14 having a region 13 on the surface of which noble metal particles are formed into an optical measuring device 12 connected to a personal computer 11 and measuring the optical characteristics of the region 13, adsorption of molecules in the region 13 is confirmed. to decide. Alternatively, the adsorption of molecules in the region 15 is determined by inserting the disc type cartridge 16 having the region 15 in which the noble metal particles are formed into the optical measuring device 17 and measuring the optical characteristics of the region 15. The optical characteristic data is loaded into the personal computer 11 and the data is analyzed. Figure 3 shows the structure of the noble metal particles. A single layer of fine particles 42 such as polymer, SiO ₂ , TiO ₂ is formed on the substrate 41, and gold, silver,
By depositing or sputtering a noble metal such as copper or platinum, hat-shaped fine particles 43 such as gold, silver, copper or platinum can be formed on the fine particles 42 (Japanese Patent Laid-Open No. 11-1703). The formation of the noble metal fine particles causes the substrate to exhibit a remarkable color development (Japanese Patent Laid-Open No. 10-339808). This coloring phenomenon occurs when light in a part of the wavelength band is absorbed when white light is reflected. Since the absorption peak wavelength of the noble metal fine particles depends on the refractive index of the surface, it can be used as a principle for detecting a reaction in which the refractive index on the surface changes (Japanese Patent Laid-Open No. 11-326193). It can be used as a biosensor by modifying it with a biomolecule having a selective adsorption ability (Japanese Patent Application Laid-Open No. 2-212058).
000-55920). This patent application uses precious metal fine particles in the form of a cartridge in order to improve the convenience of the biosensor. FIG. 4 shows various cartridge configurations. On the surface of the rectangular cartridge 51, there is a region 52 in which noble metal fine particles are formed, and the region 52 is divided into a plurality of regions. Noble metal fine particles having different characteristics are formed in each region. For example, noble metal fine particles having different sizes are formed in the regions 53 and 54, and different antibodies are bound to the fine particles. On the surface of the disc type cartridge 55, there is a region 56 in which fine metal particles are formed.
6 is divided into a plurality of areas. Noble metal fine particles having different characteristics are formed in each region. For example, different noble metal fine particles are formed in the regions 57 and 58, and different DNAs are bound to each of them. On the surface of the rod-shaped cartridge 59, there is a region 60 in which the noble metal fine particles are formed, and the region 60 is divided into a plurality of regions. Noble metal fine particles having different characteristics are formed in each region. For example, the noble metal fine particles having different sizes are formed in the regions 61 and 62, and different proteins are bound to the fine particles.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiment 1 FIG. 5 shows an embodiment of the usage form of the present invention. In the gold fine particle formation region 72 on the surface of the cartridge 71, a specific binding reaction of biomolecules is performed. After completion of the binding reaction, the cartridge 7
1 is inserted into the optical measuring device 73. The optical measuring device 73 includes an optical detecting head 74. Optical detection head 74
Has a light source 75 and an optical detector 76, and the optical detector 76 detects the reflected light from the light source 75. The optical detection head 74 is held by an arm 77, and the optical detection head 74 can be moved by being driven by an actuator 78. As a result, the gold fine particle formation region 72
It is possible to detect the optical characteristics of any region of the. Embodiment 2 FIG. 6 shows an embodiment of the usage form of the present invention. A specific binding reaction of biomolecules is performed in the gold fine particle formation region 82 on the surface of the cartridge 81. After completion of the binding reaction, the cartridge 8
1 is inserted into the optical measuring device 83. The optical measuring device 83 includes an optical detecting head 84. Optical detection head 84
Has a light source 85 and an optical detector 86, and the optical detector 86 detects the reflected light from the light source 85. The cartridge 81 is held by the arm 87, and the actuator 8
By driving with 8, the cartridge 81 can be moved. As a result, it is possible to detect the optical characteristics of any area of the gold fine particle forming area 82. Embodiment 3 FIG. 7 shows an embodiment of the usage form of the present invention. A fine silver particle region 92 is formed on the surface of the rectangular cartridge 91. A reference region 93 is formed adjacent to the region 92. The surface of the silver fine particles in the reference area 93 is treated so that a biomolecule binding reaction does not occur. Square cartridge 9
When a biomolecule binding reaction is caused to occur on one surface, the reflection spectrum 94 changes with adsorption in the region 92, but no adsorption occurs in the reference region 93, so the reference reflection spectrum 9
5 does not change. By using the reference reflection spectrum 95, the change amount 96 of the reflection spectrum in the region 92
Can be asked. Further, the reflection spectrum 97 before the biomolecule adsorption reaction is stored in the storage device of the personal computer 98 in advance and compared with the reflection spectrum 99 after the biomolecule adsorption, and the variation amount 100 of the reflection spectrum in the region 92 can be obtained. Embodiment 4 FIG. 8 shows an embodiment of the use form of the present invention. The cartridge 111 has a region 112 in which fine gold particles are formed.
In order to perform the biomolecule binding reaction in the region 112 with the cartridge 111 inserted in the optical measuring device 113,
A channel is formed in the cartridge 111, and the sample can be introduced into the region 112 by using the sample supply device 114. That is, when a sample is injected into the sample injection port 115 provided in the sample supply device 114, the small pump 1
16, valve 117, tube 118, injection channel 119
The sample is introduced into the region 112 via. The excess sample is returned to the sample supply device via the discharge flow path 120 and the tube 121. The biomolecule adsorption reaction in the region 112 is detected by the optical head 122. Embodiment 5 FIG. 9 shows an embodiment of the usage form of the present invention. When the cartridge is inserted into the optical measuring device 131, various types of cartridges can be supported by using the adapter. Adapter 1 for square cartridge 132
By inserting into the optical measuring device 131, the shape can be adapted to the shape of the cartridge insertion port 134 of the optical measuring device 131. In addition, the disk type cartridge 135 is attached to the adapter 1
It is also possible to insert it at 36. It is also possible to insert and use a plurality of rod-shaped cartridges 137 in the same adapter 138. Embodiment 6 FIG. 10 shows an embodiment of the usage form of the present invention. A magnetically storable area 142 is formed on the surface of the cartridge 141.
Exists. The cartridge 141 is replaced with the optical measuring device 143.
After being used and written in, the used information 144 is written in the area 142 to prevent reuse. This prevents errors due to cross contamination and the like.

[0006]

According to the present invention, it is possible to provide a simple sensor and a detection device capable of detecting the presence or absence of a subject with high sensitivity.

[Brief description of drawings]

FIG. 1 is a diagram showing an embodiment of a device configuration of the present invention.

FIG. 2 is a diagram showing a device configuration and a principle of a conventional technique.

FIG. 3 is a view showing noble metal fine particles on the sensor of the present invention.

FIG. 4 is a diagram showing an example of a cartridge having a plurality of biomolecule detection regions.

FIG. 5 is a diagram showing an example of measuring a cartridge surface by scanning an optical detection head.

FIG. 6 is a diagram showing an example of measuring the surface of a cartridge by driving the cartridge.

FIG. 7 is a diagram showing how to take a reference when measuring a reflection spectrum.

FIG. 8 is a view showing an example of an apparatus for supplying a sample to a cartridge inserted in an optical measuring device.

FIG. 9 is a diagram showing an example of using an adapter corresponding to cartridges of different shapes.

FIG. 10 is a diagram showing an example of a method of marking a used cartridge.

[Explanation of symbols]

11: personal computer, 12: optical measuring device, 13: area in which precious metal fine particles are formed, 14: rod-shaped cartridge, 1
5: Area in which precious metal fine particles are formed, 16: Disk type cartridge, 17: Optical measuring device, 21: High refractive index carrier, 22: Thin film of free electron metal, 23: Molecular recognition layer, 2
4: monochromatic parallel light, 25: light source, 26: incident light, 27:
Specular reflection light, 28: detector, 29: resonance incident angle, 30:
Incident angle dependent reflected light intensity, 31: new resonance incident angle, 31: reflected light intensity, 41: substrate, 42: fine particle layer,
43: Noble metal fine particles, 51: Square cartridge, 52:
Area in which fine metal particles are formed, 53: area, 54: area, 55: disk type cartridge, 56: area in which fine metal particle is formed, 57: area, 58: area, 59:
Rod-shaped cartridge, 60: area on which precious metal fine particles are formed, 61: area, 62: area, 71: cartridge, 7
2: gold fine particle forming region, 73: optical measuring device, 74: optical detecting head, 75: light source, 76: optical detector, 77:
Arm, 78: actuator, 81: cartridge,
82: Gold fine particle forming region, 83: Optical measuring device, 84:
Optical detection head, 85: light source, 86: optical detector, 8
7: Arm, 88: Actuator, 91: Square cartridge, 92: Silver fine particle forming region, 93: Reference region, 9
4: reflection spectrum, 95: reference reflection spectrum, 9
6: change amount of reflection spectrum, 97: reflection spectrum,
98: personal computer, 99: reflection spectrum, 100: change amount of reflection spectrum, 111: cartridge, 112:
Gold fine particle formation region, 113: Optical measuring device, 114: Sample supply device, 115: Sample injection port, 116: Small pump, 117: Valve, 118: Tube, 119: Injection flow path, 120: Discharge flow path, 121: Tube, 122:
Optical head, 131: Optical measuring device, 132: Square cartridge, 133: Adapter, 134: Cartridge insertion port, 135: Disk type cartridge, 136: Adapter, 137: Rod cartridge, 138: Adapter,
141: cartridge, 142: magnetic recording area, 14
3: Optical measuring device, 144: Used information.

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Claims (7)

(57) [Claims] 1. A noble metal-coated dielectric fine particle is provided with an insertion port for inserting a cartridge formed on the surface thereof, and an optical measuring device for measuring the optical characteristics of the cartridge surface, wherein the cartridge comprises: The surface of the cartridge is divided into a plurality of regions, and each region is composed of fine particles having different optical characteristics or fine particles modified with different biomolecules. Then, the biomolecule detecting device can be inserted while being accommodated in an adapter adapted to the shape of the insertion port. 2. The biomolecule detecting device according to claim 1, further comprising means for scanning the detection probe light on the area. 3. The biomolecule detecting device according to claim 3, wherein the means for scanning the detection probe light over the area is means for moving the cartridge with respect to the detection probe light whose position is fixed. . 4. The biomolecule detecting device according to claim 1, further comprising means for irradiating the plurality of regions at the same time to measure optical characteristics. 5. The biomolecule detecting device according to claim 1, further comprising means for marking the cartridge optically, magnetically or mechanically. 6. An insertion port for inserting a cartridge in which a biomolecular reaction has been completed, and a means for comparing the detected optical characteristic with optical characteristic data stored in advance in a recording means connected to an optical measuring device. The biomolecule detecting device according to claim 1. 7. A biomolecule according to claim 1, further comprising: an insertion opening for inserting a cartridge in which the biomolecule reaction has been completed, and a means for comparing with an optical characteristic of a reference area formed on the surface of the cartridge. Detection device.