JP2003083974A - Biochemical analyzing unit and method for manufacturing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To arrange a specific bonding substance to a biochemical analyzing unit in high density. SOLUTION: Metal oxide particles are dispersed in plastics in order to lower light transmissivity to constitute a substrate 2 and holes 3 are formed to the substrate 2 in high density. Aliphatic polyamide is dissolved in a solution to be injected in the holes 3. When the substrate 2 is immersed in a coagulation bath to be dried, aliphatic polyamide is made porous to become a porous material 4. After the specific bonding substance originating from a living body is fixed to the porous material 4, a substance originating from the living body to which a radioactive label substance is applied is dropped on the porous material 4. When the specific bonding substance and the substance originating from the living body are hybridized, radioactive rays are discharged from the holes 3. Even if radioactive rays are emitted from a large number of the holes, it is suppressed that radioactive rays are scattered to be mixed because the light transmissivity of the substrate 2 is low. Noise is suppressed to enable biochemical analysis of high accuracy.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for manufacturing a biochemical analysis unit and a biochemical analysis unit. More specifically, spots of a specific binding substance capable of specifically binding to a substance of biological origin and having a known base sequence, base length, composition, etc. are formed at high density on the unit surface, The specific binding substance of, by hybridizing a substance of biological origin labeled with a radioactive labeling substance, the biochemical analysis unit obtained by selective labeling is brought into close contact with the stimulable phosphor layer, The stimulable phosphor layer is exposed to a radioactive labeling substance, the stimulable phosphor layer is irradiated with excitation light, and the stimulable light emitted from the stimulable phosphor layer is photoelectrically detected to emit radiation. Even when an image is generated and a substance derived from a living body is analyzed, noise caused by scattering of electron beams emitted from a radiolabeled substance can be prevented from being generated in a radiation image, and the substance derived from a living body can be prevented. Can be specifically bound to and is a base Length of the column and the base, a spot of known specific binding substances such as composition unit surface,
A labeling substance and / or fluorescence which is formed at a high density and causes chemiluminescence by being brought into contact with a chemiluminescent substrate in addition to the radiolabeling substance or in place of the radiolabeling substance in a spot-like specific binding substance By hybridizing a substance derived from a living body labeled with a substance, and photoelectrically detecting chemiluminescence and / or fluorescence emitted from the biochemical analysis unit obtained by selectively labeling,
When a substance derived from a living body is analyzed by generating a chemiluminescence image and / or a fluorescence image, a labeling substance that causes chemiluminescence by contact with a chemiluminescence substrate and / or
Alternatively, a method for manufacturing a biochemical analysis unit and biochemistry capable of preventing generation of noise resulting from chemiluminescence emitted from a fluorescent substance and / or scattering of fluorescence in a chemiluminescence image and / or fluorescence image It relates to an analysis unit.

[0002]

2. Description of the Related Art When a radiation is irradiated, the energy of the radiation is absorbed, stored and recorded, and then excited by using an electromagnetic wave of a specific wavelength range. A photostimulable phosphor having the property of emitting a stimulating amount of light is used as a radiation detection material, and a substance having a radioactive label is administered to an organism, and then the organism or tissue of the organism is treated. A portion of the sample is used as a sample, and this sample is overlapped with a stimulable phosphor sheet provided with a stimulable phosphor layer for a certain period of time to store and record radiation energy in the stimulable phosphor. , Scanning the stimulable phosphor layer with electromagnetic waves to excite the stimulable phosphor and photoelectrically detecting the stimulable light emitted from the stimulable phosphor to generate a digital image signal. Image processing, CRT On a recording material such as a display unit or on a photographic film, configured autoradiographic image detecting system to reproduce an image has been known (for example, Kokoku 1-60784 and JP Kokoku 1-
No. 60782, Japanese Patent Publication No. 4-3952, etc.).

An autoradiographic image detection system using a stimulable phosphor sheet as an image detection material is
Unlike the case of using a photographic film, not only a chemical process called developing process is unnecessary, but also by performing image processing on the obtained image data, an image is reproduced as desired or a computer is used. It has an advantage that quantitative analysis can be performed.

On the other hand, there is known a fluorescence image detection system which uses a fluorescent dye as a labeling substance instead of the radioactive labeling substance in the autoradiography image detection system. According to this fluorescence image detection system, by reading the fluorescence image, the gene sequence, the expression level of the gene, the metabolism, absorption and excretion pathways and states of the administered substance in experimental mice, the separation, identification of the protein, or the molecular weight , Characteristics can be evaluated, and, for example, a solution containing a plurality of types of protein molecules to be electrophoresed is electrophoresed on a gel support,
An image is generated by staining the electrophoresed protein by immersing the gel support in a solution containing a fluorescent dye, exciting the fluorescent dye with excitation light, and detecting the resulting fluorescence, The position and quantitative distribution of protein molecules on the gel support can be detected. Alternatively, by western blotting, at least a part of the electrophoresed protein molecule is transferred onto a transfer support such as nitrocellulose, and the antibody that specifically reacts with the target protein is labeled with a fluorescent dye. By associating the prepared probe with a protein molecule, selectively labeling a protein molecule that binds only to an antibody that specifically reacts, by exciting light, exciting a fluorescent dye, and detecting the resulting fluorescence,
Images can be generated to detect the location and quantitative distribution of protein molecules on the transfer support. In addition, after adding a fluorescent dye to a solution containing a plurality of DNA fragments to be electrophoresed, the plurality of DNA fragments are electrophoresed on a gel support, or on a gel support containing a fluorescent dye. , Electrophores multiple DNA fragments, or
A plurality of DNA fragments are electrophoresed on a gel support, and then the gel support is immersed in a solution containing a fluorescent dye to label the electrophoresed DNA fragments, and the DNA is excited by excitation light to fluoresce. An image is generated by exciting the dye and detecting the generated fluorescence to detect the distribution of DNA on the gel support, or after a plurality of DNA fragments are electrophoresed on the gel support. , Denature DNA (de-naturatio
n) Then, by a Southern blotting method, at least a part of the denatured DNA fragment is transferred onto a transfer support such as nitrocellulose, and the DNA or RNA complementary to the target DNA is labeled with a fluorescent dye. DNA that is complementary to the probe DNA or probe RNA by hybridizing the probe prepared by
By selectively labeling only the fragments, exciting the fluorescent dye with excitation light, and detecting the resulting fluorescence, an image is generated and the distribution of the target DNA on the transfer support is detected. be able to. Furthermore, DNA complementary to the DNA containing the target gene labeled with a labeling substance
A probe is prepared, hybridized with DNA on a transcription support, and an enzyme is bound to a complementary DNA labeled with a labeling substance, and then contacted with a fluorescent substrate to fluoresce the fluorescent substrate. Generating an image by changing the substance to a substance, exciting the generated fluorescent substance with excitation light, and detecting the generated fluorescence, and detecting the distribution of the target DNA on the transfer support. You can also This fluorescence image detection system has an advantage that a gene sequence or the like can be easily detected without using a radioactive substance.

Similarly, a substance derived from a living body such as a protein or a nucleic acid is immobilized on a support and is selectively labeled with a labeling substance which causes chemiluminescence by contacting with a chemiluminescent substrate. A selectively labeled biological substance is brought into contact with a chemiluminescent substrate, and chemiluminescence in the visible light wavelength region generated by the contact between the chemiluminescent substrate and the labeled substance is photoelectrically detected to obtain a digital image signal. Chemiluminescence detection for generating information, reproducing the chemiluminescence image on display means such as CRT or recording material such as photographic film by performing image processing, and obtaining information on substances of biological origin such as genetic information The system is also known.

Furthermore, in recent years, hormones, etc. have been placed at different positions on the surface of a unit such as a slide glass plate and a porous membrane.
Can specifically bind to substances of biological origin, such as tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA (complementary DNA synthesized using mRNA as a template), DNA, RNA, and A specific binding substance with known base sequence, base length, composition, etc. is dropped using a spotter device to form a large number of independent spots, and then hormones, tumor markers, enzymes, antibodies , Antigens, abzymes, other proteins, nucleic acids, c
A substance derived from a living body, such as DNA, DNA, or mRNA, which is collected from the living body by extraction or isolation, or which is further subjected to a chemical treatment, a chemical modification, or the like, such as a fluorescent substance or a dye. A microarray hybridized with a substance labeled with a labeling substance is irradiated with excitation light, and light such as fluorescence emitted from a labeling substance such as a fluorescent substance or a dye is photoelectrically detected to obtain a substance of biological origin. A microarray image detection system has been developed to analyze the. According to this microarray image detection system, a large number of spots of specific binding substances are formed at high density at different positions on the surface of a unit such as a slide glass plate or a porous membrane, and the biological substance labeled with a labeling substance is derived from the living body. By hybridizing the substance of 1), it is possible to analyze the substance of biological origin in a short time.

[0007] In addition, hormones, tumor markers, enzymes, antibodies,
Antigen, abzyme, other protein, nucleic acid, cDNA
A specific binding substance, such as A, DNA, or RNA, which can be specifically bound to a substance of biological origin and whose base sequence, base length, composition, etc. is known, is dropped using a spotter device. Form a large number of independent spots, and then hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, m
A substance derived from a living body, such as RNA, which is collected from a living body by extraction, isolation, or the like, or which has been further subjected to a treatment such as a chemical treatment or a chemical modification, which is labeled with a radiolabeled substance, is hybridized. The soybean macroarray was brought into close contact with the stimulable phosphor sheet having the stimulable phosphor layer containing the stimulable phosphor, and the stimulable phosphor layer was exposed to light. A macroarray image detection system using a radiolabeled substance that irradiates the phosphor layer with excitation light and photoelectrically detects the photostimulable light emitted from the photostimulable phosphor layer to analyze a substance derived from a living body is also available. Being developed.

[0008]

However, in the macroarray image detection system using the radiolabeled substance, when the photostimulable phosphor layer is exposed, it is formed on the unit surface such as a porous film. Since the radioactive energy of the radiolabeled substance contained in the spot is very large, the electron beam emitted from the radiolabeled substance is scattered inside the unit such as the porous membrane and is exposed by the radiolabeled substance contained in the adjacent spot. The electron beam emitted from the radiolabeled substance that is incident on the region of the stimulable phosphor layer to be scattered is scattered, and the electron beams emitted from the radiolabeled substances contained in the adjacent spots are mixed with each other to give a bright emission. When it enters the region of the photostimulable phosphor layer, as a result, noise is generated in the radiation image generated by photoelectrically detecting stimulated emission, and the radiation dose of each spot is quantified. Te, when analyzing the substance derived from a living organism,
There is a problem that the quantification deteriorates, and particularly when the spots are formed close to each other to increase the density, a remarkable deterioration of the quantification is recognized.

In order to prevent the noise caused by the scattering of the electron beam emitted from the radio-labeled substance contained in the adjacent spots and solve this problem, the distance between the adjacent spots must be increased. You will need to
There is a problem that the density of the spots is lowered and the inspection efficiency is lowered.

Furthermore, in the field of biochemical analysis, hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins formed in spots at different positions on the surface of a unit such as a porous membrane, Nucleic acid, c
In addition to radiolabeled substances, specific binding substances that can specifically bind to substances of biological origin, such as DNA, DNA, and RNA, and whose base sequence, base length, and composition are known,
A substance of biological origin labeled with a labeling substance and / or a fluorescent substance that produces chemiluminescence when brought into contact with a chemiluminescent substrate is hybridized and selectively labeled. After exposing the layer, or prior to exposure of the stimulable phosphor layer with a radiolabeled substance, contact with a chemiluminescent substrate,
Photoelectrically detecting chemiluminescence in the visible light wavelength region generated by contact between the chemiluminescent substrate and the labeling substance, and / or
It is also required to irradiate excitation light and photoelectrically detect fluorescence emitted from a fluorescent substance to analyze a substance derived from a living body, but in such a case, chemiluminescence or fluorescence emitted from a spot is also required. Are scattered in the unit such as the porous membrane,
Alternatively, chemiluminescence or fluorescence emitted from a spot is scattered and mixed with chemiluminescence or fluorescence emitted from an adjacent spot, and as a result, a chemiluminescence image generated by photoelectrically detecting chemiluminescence and / or Alternatively, there is a problem that noise is generated in a fluorescent image generated by photoelectrically detecting fluorescent light.

The present invention is intended to solve the above problems, and an object of the present invention is to provide a method for efficiently producing a biochemical analysis unit capable of performing highly accurate biochemical analysis while suppressing the generation of noise. To do.

[0012]

In order to achieve the above object, according to the invention of claim 1, a substrate formed of a material having a property of attenuating radiation and / or light and having a plurality of holes is formed. In the method for manufacturing the biochemical analysis unit, wherein the plurality of holes each have an absorptive region formed therein, the plurality of holes are each filled with an absorptive material to form the absorptive region. is doing.

According to the second aspect of the present invention, it is formed of a material having a property of attenuating radiation and / or light, and a plurality of holes are formed, and the absorptive regions are respectively formed in the plurality of holes. Is formed, thereby comprising a substrate having a plurality of absorptive regions formed, in the plurality of absorptive regions formed in the plurality of holes of the substrate,
At least one labeling substance selected from the group consisting of labeling substances that cause chemiluminescence when a specific binding substance having a known structure or property is dropped and brought into contact with either a radioactive labeling substance, a fluorescent substance or a chemiluminescent substrate. In the method for producing a biochemical analysis unit, wherein a substance derived from a living body labeled by is specifically bound to the specific binding substance, and the plurality of absorptive regions are selectively labeled, The holes are filled with an adsorptive material to form the adsorptive regions.

According to the third aspect of the present invention, an absorptive region is formed in each of the plurality of holes of the substrate formed of a material having a property of attenuating radiation and having a plurality of holes. , A plurality of absorptive regions formed, a substance of biological origin labeled with a radioactive labeling substance by dropping a specific binding substance capable of specifically binding to a substance of biological origin and having a known structure or characteristic Is specifically bound to the specific binding substance to selectively label the plurality of absorptive regions, and to the stimulable phosphor sheet having a stimulable phosphor layer formed thereon, The phosphor layer is opposed to the plurality of absorptive regions so that the layers are overlapped with each other, and the radioactive labeling substance contained in the plurality of absorptive regions,
Exposing the stimulable phosphor layer, irradiating the stimulable phosphor layer exposed by the radioactive labeling substance with excitation light, and a stimulable phosphor contained in the stimulable phosphor layer. A biochemical analysis unit for generating biochemical analysis data by photoelectrically detecting photostimulated light emitted from the photostimulable phosphor contained in the photostimulable phosphor layer. In the manufacturing method described above, each of the plurality of holes is filled with an adsorptive material to form the adsorptive region.

After the main component of the solvent composition of the adsorbent material is a good solvent for the adsorbent material and an adsorbent material solution containing a small amount of a poor solvent is injected into the plurality of holes of the substrate, the substrate is removed. It is preferable to fill the adsorptive material by immersing in the coagulating liquid or gradually drying. Further, the adsorbent material formed in a film shape in advance and the substrate are overlapped and pressed, and the film-shaped adsorbent material is pressed into the holes of the substrate to fill the adsorbent material. Is preferably performed.

The adsorptive material is preferably made of a porous material. Furthermore, it is preferable that the porous material is made of a material capable of forming a membrane filter.
Further, it is preferable to mix a fibrous material insoluble in the solvent of the porous material with the porous material.

When the material having the property of attenuating the radiation and / or the light transmits the radiation and / or the light through the material by a distance equal to the distance between the adjacent absorptive regions, Alternatively, it is preferable that the light energy be reduced to 1/5 or less.

It is preferable that the substrate is formed of a material selected from the group consisting of metal materials, ceramic materials and plastic materials.

The method of forming the plurality of holes in the substrate is preferably a punching method, an electric discharge machining method, a method combining photolithography and etching, or a laser ablation method. The hole may be a hole having a concave shape instead of the hole penetrating the substrate.

The size of the holes is preferably less than 5 mm ² . Further, it is preferable that the holes are formed at a density of 10 holes / cm ² or more.

[0021]

BEST MODE FOR CARRYING OUT THE INVENTION The embodiments of the present invention will be described below, but the present invention is not limited thereto. Further, terms and the like used in the following description show preferred examples of the present invention, and do not show the meaning or technical scope of the terms of the present invention.

FIG. 1 is a perspective view showing the outline of the biochemical analysis unit of the present invention. The biochemical analysis unit 1 shown in FIG. 1 includes a substrate 2 having a plurality of holes 3 and a porous material 4 filled in the holes 3 and bonded to the substrate 2. A specific binding substance 5 having a known structure or characteristic is dropped on the porous material 4 serving as the adsorptive region, and immobilized by the subsequent treatment.

The material of the substrate 2 is preferably a material that does not transmit or attenuate radiation or light in order to prevent scattering inside the biochemical analysis unit, and is preferably metal or ceramic. Also, when using a plastic that is easy to make a hole as the substrate,
It is preferred to disperse the particles within the plastic in order to further attenuate the radiation or light.

The metal includes copper, silver, gold, zinc,
Examples thereof include lead, aluminum, titanium, tin, chromium, iron, nickel, cobalt, tantalum, and alloys such as stainless steel and brass, but are not necessarily limited thereto.

Examples of the plastics include polyolefins such as polyethylene and polypropylene, acrylic resins such as polystyrene and polymethylmethacrylate, polyvinyl chloride, polyvinylidene chloride, polyvinylidene fluoride, polytetrafluoroethylene, polychlorotrifluoroethylene, polycarbonate, Polyesters such as polyethylene naphthalate and polyethylene terephthalate, aliphatic polyamides such as nylon-6 and nylon-66, silicon resins such as polyimide, polysulfone, polyphenylene sulfide, polydiphenyl siloxane, phenol resins such as novolac, epoxy resins, polyurethane, Cellulose such as cellulose acetate and nitrocellulose, copolymer such as butadiene-styrene copolymer, Although the like blending the plastic, but are not necessarily limited to.

In order to attenuate radiation or light, it is preferable to fill the above-mentioned plastic with metal oxide particles or glass fibers. Examples of the metal oxide particles include silicon dioxide, alumina, titanium dioxide, iron oxide, and oxide. Examples thereof include copper, but are not necessarily limited thereto.

Examples of the ceramic include, but are not limited to, alumina, zirconia, magnesia, and quartz.

The meaning of attenuating radiation or light means that the specific binding substance on the surface or inside of the porous material filled in the holes 3 of the substrate 2 and the radiation or light emitted from the bound labeling substance is emitted from the holes. The strength of penetrating the substrate wall and reaching the adjacent holes is preferably 1/5 or less, more preferably 1/10 or less.

In order to effectively shield radiation such as an electron beam from a radiolabeled sample, the average density of the substrate 2 is generally 0.6 g / cm ³ or more, preferably 1 to 20 g / cm ^{3. It is in} the range of ³ , particularly preferably 2
It is in the range of 10 g / cm ³ . Since the penetration distance of the electron beam is inversely proportional to the density, radioactive material is 32P, 33P, 35
Common radioisotopes (RI) such as S, 14C, etc.
Then, by setting the average density of the substrate 2 within this range, the electron beam from the RI of the sample to be fixed in each hole 3 is shielded by the partition wall of the substrate 2 to transmit the electron beam, It is possible to prevent deterioration of the resolution of the radiation image due to scattering.

The thickness of the substrate 2 is generally 50 to 10
Is in the range of 00 μm, preferably 100 to 500 μm
Is in the range.

The area (size) of the opening of the hole 3 formed in the substrate 2 is generally less than 5 mm ² , preferably less than 1 mm ² , in order to increase the density of the hole 3, Less than 3 mm ² is more preferable, and 0.01 mm ²
Less than is more preferred. And preferably 0.001
mm ² or more.

The pitch P1 of the holes 3 (the distance from the center to the center of two adjacent holes) P1 is generally in the range of 0.05 to 3 mm, and the distance between the holes 11 (from the end to the end of the two adjacent holes). The shortest distance to the part) L1 is generally 0.01 to
It is in the range of 1.5 mm. The number (density) of the holes 3 is generally 10 holes / cm ² or more, preferably 100 holes / c.
m ² or more, more preferably 500 / cm ² or more, still more preferably 1000 / cm ² or more. And it is preferably 100,000 pieces / cm ² or less, and particularly preferably 10,000 pieces / cm ² or less. It should be noted that all of them are not necessarily provided at equal intervals as shown in FIG. 1, and a plurality of holes 3 may be provided for each block divided into some blocks (units). .

As a method of forming a plurality of holes 3 in the substrate 2, punching by punching with a pin can be mentioned. In order to improve efficiency, as shown in FIG. 2, it is preferable to dispose a plurality of pins 9 at a distance that is an integral multiple of the distance between the holes and punch a plurality of holes 3 by one punch.

As a method for forming a plurality of holes 3 in the substrate 2, a discharge electrode having the same electrode arrangement pattern as the hole pattern is brought close to a grounded substrate in an electrically insulating fluid such as oil or air. Then, by applying a high voltage in a pulsed manner to the discharge electrode, the heat generated by the generated discharge elevates the electric discharge machining for volatilizing the substrate.

The plurality of holes 3 in the substrate 2 may be provided by photolithography and etching. Figure 3 (a)
As shown in FIG. 1, first, a photosensitive material is applied on the support 10 to form a coating film 8, and then a photolithographic mask material 7 having a desired hole pattern is formed on the coating film 8 by using light. By photolithography to transfer, coating film 8 around the hole
Cure. After that, the coating film 8 is immersed in an organic solvent, and a plurality of holes are formed in the coating film 8 by etching to elute and remove the non-irradiated portion in the organic solvent. The substrate 2 is peeled off to obtain a substrate 2 having a large number of holes 3 as shown in FIG. 3 (b). Examples of the support include polyethylene, polypropylene, polyethylene terephthalate, and polytetrafluoroethylene, but the present invention is not limited thereto.

An ultraviolet curable composition is preferably used as the photosensitive material. The ultraviolet curable composition,
It consists of a photopolymerization initiator and an ultraviolet curable resin material. The photopolymerization initiator may be any substance containing a source that initiates photopolymerization. For example, a hydrogen abstraction type initiator (eg, benzophenone type initiator) or a radical cleavage type initiator (eg,
Acetophenone-based initiators and triazine-based initiators) can be used. Examples of the ultraviolet curable resin raw material used in the present invention include acrylic acid esters (eg, ethyl acrylate, butyl acrylate, 2-ethylhexyl acrylate), methacrylic acid esters (eg, methyl methacrylate, methacrylic acid). Ethyl acid ester, butyl methacrylate, ethylene glycol dimethacrylate) Ester of polyhydric alcohol and (meth) acrylic acid (eg, ethylene glycol di (meth) acrylate, 1,4-dichlorohexane diacrylate, pentaerythritol tetra (meth)) Acrylate), pentaerythritol tri (meth) acrylate, trimethylolpropane tri (meth) acrylate, trimethylolethane tri (meth) acrylate, dipentaerythritol tetra (meth) acrylate, dipentaerythritol Pointer (meth) acrylate, pentaerythritol hexa (meth) acrylate, 1,2,3-cyclohexane tetramethacrylate, polyurethane polyacrylate, not but polyester polyacrylate) can be cited as being limited thereto. These may be used alone or in plural kinds.

As the organic solvent for etching, any organic solvent that dissolves the ultraviolet curable composition is used. Examples include, but are not limited to, ketones such as acetone and ethyl methyl ketone.

In order to easily remove the non-irradiated portion by etching, it is preferable to perform etching while irradiating the substrate with ultrasonic waves in an etching solution.

When a metal is used for the substrate 2, a plurality of holes may be provided by electrolytic etching. First, a resist applied on a metal plate is exposed with a photomask pattern in which a hole pattern to be produced is drawn and developed. For example, in a strong acid solution such as sulfuric acid, hydrofluoric acid or phosphoric acid, the metal plate is used as an anode and platinum is used as a cathode to perform electrolysis to perform holes in the metal plate by electrolytic etching, and then the resist remaining on the surface of the metal plate is removed. .

As another method of forming a plurality of holes in the substrate, there is a method of irradiating the substrate with a high-power laser beam such as an excimer laser or a YAG laser and heating the substrate in the irradiated portion to volatilize it. . An array of holes is formed by scanning the laser beam along the surface of the substrate. Further, the holes 3 may be recessed holes (holes) in addition to the holes penetrating the substrate 2.

In the present invention, a porous material or a fibrous material is preferably used as the absorptive material forming the absorptive region. Further, the absorptive region can be formed by using the porous material and the fiber material together. In the present invention, the porous material used for forming the adsorptive region may be either an organic material or an inorganic material, or may be an organic / inorganic composite.

In the present invention, the organic porous material used to form the adsorptive region is not particularly limited, but a carbon porous material such as activated carbon or a porous material capable of forming a membrane filter. Are preferably used. A polymer soluble in a solvent is preferably used as the porous material capable of forming the membrane filter. Examples of the polymer include cellulose derivatives (eg, nitrocellulose, regenerated cellulose, cellulose acetate, cellulose acetate, cellulose butyrate acetate, etc.), aliphatic polyamides (eg, nylon 6, nylon 6,6, nylon 4,10, etc.), Polyolefins (for example, polyethylene, polypropylene, etc.), chlorine-containing polymers (for example, polyvinyl chloride, polyvinylidene chloride, etc.), fluororesins (for example, polyvinylidene fluoride, polytetrafluoride, etc.), polycarbonate, polysulfone, Examples include alginic acid and its derivatives (eg, alginic acid, calcium alginate, alginic acid / polylysine polyion complex, etc.), collagen, etc., and copolymers or complexes (mixtures) of these polymers should also be used. It can be.

The fiber material for forming the absorptive region is not particularly limited, but preferably the above-mentioned cellulose derivatives, aliphatic polyamides and the like are mentioned.

In the present invention, the inorganic porous material used to form the absorptive region is not particularly limited, but is preferably a metal (for example, platinum, gold,
Examples thereof include iron, silver, nickel, aluminum, etc., oxides of metals (eg, alumina, silica, titania, zeolite, etc.), metal salts (eg, hydroxyapatite, calcium sulfate, etc.), and composites thereof.

Further, in order to enhance the strength of the porous material 4 as the adsorptive region, a fibrous material insoluble in the solvent of the porous material 4 may be mixed with the porous material 4.
The fibrous material may be any material as long as it is a material insoluble in the solvent of the porous material 4. For example, cellulose,
Glass fibers and metal fibers are used. These may be used alone or in plural kinds.

FIG. 4 shows a schematic view of an embodiment of a method for continuously injecting the dope (solution containing the porous material) 4 into the holes 3 of the substrate 2. The dope 4 is formed on the substrate 2 above the substrate 2 which is continuously conveyed.
A casting die 11 is arranged so that the dope 4 is continuously injected into the substrate 2. Excessive dope 4 overflowing from the holes is scraped off by a blade 12 fixed above and below the substrate 2. The substrate 2 into which the dope is injected is immersed in a coagulation bath (coagulation liquid) 13 composed of a mixed solution of a good solvent and a poor solvent of a porous material or a poor solvent solution of a porous material, and each of the substrates 2 is A porous film is formed in the holes 3. The substrate on which the porous film is formed is immersed in the water bath 14 for cleaning, and then carried to the drying step.

The concentration of the dope 4 is generally 5 to 35.
It is in the range of wt%, preferably in the range of 10 to 30 wt%. When the concentration is lower than 5% by weight, the strength of the porous structure such as the porous material 4 and the porous membrane 13 is not sufficient, while when the concentration is higher than 35% by weight, the porosity of the porous structure decreases. Then, the water permeability to the probe molecule solution becomes so small that it has no substantial meaning.

FIG. 5 shows a schematic view of one form of a method for intermittently injecting the dope into the holes 3 of the substrate 2.
A dispenser 17 for injecting the dope 4 into the holes 3 of the substrate 2 is arranged above the substrate 2 which is continuously or intermittently transported, and the dispenser 17 intermittently doespe the dope into each hole 3. inject.

As a method of forming porosity in each hole of the substrate, by supplying a constant temperature and humidity to the substrate into which the dope is injected at a constant wind speed, and gradually evaporating the solvent, A method that causes phase separation or gelation may be used.

As a method of removing the dope overflowing from the holes, a method of installing a suction device above and below the substrate 2 and sucking excess dope may be used.

In order to accelerate the permeation of the specific binding substance into the porous material, it is preferable to make the surface of the porous material hydrophilic by an electric discharge treatment. When the substrate is a conductive material such as metal, a method of grounding the substrate so that an electrode to which an AC high voltage is applied is opposed to the substrate can be mentioned. When the substrate is an insulating material such as plastic or ceramic, there is a method in which the substrate is placed on a grounded conductive material and the electrodes to which an AC high voltage is applied are opposed to the substrate.

In order to accelerate the permeation of the specific binding substance into the porous material, it is preferable to impregnate the porous material with a surfactant. As the surfactant, any of anionic, cationic and fluorine-based surfactants may be used. Specifically, sodium dodecylbenzenesulfonate, saponin, sodium p-tert-octylphenoxyethoxyethylsulfonate, nonylphenoxypolyethoxyethanol, JP-A-62-17
0950, US Pat. No. 5,380,644,
Fluorine-based surfactants described in JP-A-63-188135 and the like, and polyalkylene oxides and anionic surfactants described in JP-A-6-301140 and the like can be mentioned.

The static contact angle of water on the surface of the porous material is preferably 60 ° or less, more preferably 50 ° or less by a method of accelerating the permeation of the specific binding substance into the porous material. is there.

In order to increase the strength of the porous material with which the substrate is filled, a plurality of different porous materials may be sequentially injected and solidified in the plurality of holes of the substrate.

The porous material filled in the holes of the substrate is
As a method for adhering to the substrate, there is a method in which an adhesive such as epoxy is thinly applied to the substrate to adhere the substrate and the porous material, and more preferable methods include the following methods. . When the substrate is a metal, first, a metal oxide is formed on the surface by anodizing treatment or the like. The anodizing treatment is a method of forming an oxide on the surface by using a direct current in a solution of sulfuric acid, phosphoric acid and chromic acid and using the substrate metal as an anode. When the substrate is plastic, the metal oxide particles are dispersed as described above. When the substrate is ceramic, it may be a metal oxide as described above. Next, a silane or titanate coupling agent having an alkoxide group at the terminal and an amino group or a carboxyl group at the other terminal is applied to the substrate and then heated to 50 ° C. or higher to give a substrate. The hydroxyl group on the surface of the metal oxide on the surface is reacted with the alkoxide group of the coupling agent. Subsequently, the pores of the substrate can be filled with the porous material to bond the coupling agent bonded to the surface of the substrate and the porous material. As a method of applying the coupling agent to the substrate, by immersing the substrate in a coupling agent solution,
Examples of the method include spraying the coupling agent solution on the substrate, but are not limited thereto.

FIG. 6 shows a method of bonding the coupling agent 32 to the surface of the hole 3 of the substrate 2 when the substrate 2 is made of metal. First, as shown in FIG. 6A, a metal oxide layer 31 is formed on the surface of the holes 3 of the metal substrate 2 by anodization, and then the substrate 2 is immersed in a solution of the coupling agent 32. After immersion, as shown in FIG. 6B, after applying the coupling agent 32 to the surface of the hole 3 of the substrate 2,
The substrate 2 is heated at 50 ° C. or higher, as shown in FIG.
The metal oxide layer 31 formed on the surface of the hole 3 is chemically bonded to the coupling agent 32.

FIG. 7 shows a method of bonding the coupling agent 32 to the surface of the hole 3 of the substrate 2 when the substrate 2 is made of plastic. As shown in FIG. 7A, the metal oxide particles 33 are previously dispersed inside the substrate 2, and the metal oxide particles 33 are present on the surfaces of the holes 3 of the substrate 2. The substrate 2 is immersed in a solution of the coupling agent 32, and as shown in FIG.
After applying the coupling agent 32, it is heated at 50 ° C. or higher to chemically react the metal oxide particles 33 formed on the surface of the hole 3 of the substrate 2 with the coupling agent 32 as shown in FIG. 7C. To combine. When the porous material is an aliphatic polyamide, the coupling agent is preferably a silane coupling agent or a titanate coupling agent having an amino group at the terminal.

FIG. 8 shows a schematic view of an embodiment of a method of pressing the porous film 21 and the substrate 2 in a superimposed manner and press-fitting the porous film 21 into the holes 3 of the substrate 2. ing. The porous membrane 21 is formed by casting or coating the dope on a support, immersing it in a poor solvent of the polymer of the porous membrane, or a mixed solution of a good solvent and a poor solvent, followed by washing with water and drying, or dope. It is separately prepared by casting or coating on a support and then gradually drying. As shown in FIG. 8 (a), the substrate 2 having the holes 3 formed therein and the porous film 21 are overlapped, fed between a press roll 22 and a backup roll 23, and pressed, whereby ), The porous film 21 is pressed into the holes 3 of the substrate 2. In this case, when the porous film 21 is softened by a method of heating the press roll 22 and the backup roll 23,
It is possible to easily press fit the substrate 2 into the hole 3.

The average density of the porous structures such as the porous material 4 and the porous membrane 21 is set in order to effectively immobilize a nucleic acid or a fragment thereof, or a specific binding substance (probe molecule) such as a synthetic oligonucleotide. Are generally 1.
0 g / cm ³ or less, preferably 0.5 g / cm ³
Or less, and preferably 0.1 g / cm ³ or more (provided that the average density of the porous structure is smaller than the average density of the substrate 2).

Further, the above-mentioned porous structure generally has a porosity (volume ratio) in the range of 10 to 90%, and the average pore diameter of the micropores constituting the void is in the range of 0.1 to 50 μm.

At least one of the front surface and the back surface of the porous material 4 serving as the absorptive region is preferably recessed toward the inside of the substrate rather than the front surface or the back surface of the substrate 2, as shown in FIG. . With such a configuration, spotting of the probe molecule solution on the porous material 4 is facilitated, and once the spotted probe molecule solution flows out to the surface of the substrate 2 or onto another porous material 4. Can be prevented, and at the same time, scattering of radiation from the target molecules bonded and fixed to the porous material 4 can be effectively prevented.

In the present invention, the arrangement of the holes 3 in the substrate 2 and the shape of the openings of the holes 3 are not limited to the grid-like arrangement and the circular openings as shown in FIG. The arrangement, shape, etc. of can be appropriately changed.

FIG. 10 is a plan view showing an example of a variation of the arrangement of the absorptive regions. In FIG. 10, the absorptive regions 13 are arranged so as to be displaced from each other in adjacent columns.

11 and 12 are plan views showing examples of variations in the shape of the opening of the absorptive region. In FIG. 11, the opening of the absorptive region 13 is a quadrangle. In FIG. 12, the opening of region 13 is triangular.

In addition to this, the absorptive regions may be arranged at random, and the openings may be elliptical or polygonal such as hexagonal. Furthermore, it is also possible to provide a long and narrow rectangular absorptive region in a stripe shape and partition the region only in the one-dimensional direction.

As the probe molecule (specific binding substance) that can be immobilized on the adsorptive region, various polynucleotides and oligonucleotides that can be conventionally used as probe molecules for macro arrays can be used. For example, a cDNA, a part of cDNA, a polynucleotide prepared by amplification by a PCR method such as EST (“PCR
Products ”), and synthetic oligonucleotides. Further, it may be an artificial nucleic acid obtained by converting a phosphodiester bond of DNA into a peptide bond, that is, a peptide nucleic acid (PNA), or a derivative thereof. In addition, hormones, tumor markers, enzymes, antibodies,
Antigen, abzyme, other protein, nucleic acid, cDNA
All specific binding substances, such as A, DNA, and RNA, which can be specifically bound to substances of biological origin and whose base sequence, base length, composition, etc. are known, should be used as the probe molecule of the present invention. You can

Specific examples of the combination of the probe molecule and the target molecule include DNA (DNA or a fragment thereof, or oligo DNA) and DNA, and DNA and RN.
A, PNA and DNA or RNA, PNA and PNA, antigen and antibody, avidin and biotin, and the like.

As a method of dropping the probe molecule into the porous material, as shown in US Pat. No. 5,807,522, a liquid containing the probe molecule attached to a pin is used.
Examples include, but are not limited to, a spotting method of transferring to a porous material and an inkjet method of discharging droplets containing probe molecules to transfer to a porous material.

The probe molecule can be immobilized on the porous material by physical adsorption on the surface of the pores of the porous material, but more preferably by a chemical reaction or a method such as heating or ultraviolet irradiation. It is preferable to bind to.

The target molecule fixed to the pore surface of the porous material and bound to the probe molecule is, for example,
If it is labeled with one or more labeling substances selected from the group consisting of radioactive labeling substances such as 33P, fluorescent labeling substances such as fluorescent dyes, and labeling substances that emit chemiluminescence when contacted with a chemiluminescent substrate. Good. Specific binding between the specific binding substance and the labeled substance derived from a living body includes, for example, hybridization between cDNAs, antigen-antibody reaction, reaction such as receptor / ligand, but is not necessarily limited thereto.

The photostimulable phosphor for accumulating the radiation emitted from the radio-labeled substance in the present invention is capable of accumulating the energy of the radiation and is excited by an electromagnetic wave, and the energy of the accumulated radiation in the form of light. There is no particular limitation as long as it can emit light, but one that can be excited by light in the visible light wavelength range is preferable. Specifically, for example, the alkaline earth metal fluorohalide-based phosphor (Ba _1-x M ²⁺ _x ) FX: yA (here, M ²⁺ ) disclosed in JP-A-55-12145 is disclosed. Is Mg,
At least one alkaline earth metal element selected from the group consisting of Ca, Sr, Zn and Cd, X is Cl, Br
And at least one halogen selected from the group consisting of I, A is Eu, Tb, Ce, Tm, Dy, Pr, H
At least one trivalent metal element selected from the group consisting of e, Nd, Yb, and Er, x is 0 ≦ x ≦ 0.6, and y is 0 ≦ y ≦ 0.2. ), An alkaline earth metal fluorohalide-based phosphor SrFX: Z disclosed in JP-A-2-276997 (where X is Cl, Br and I).
At least one halogen selected from the group consisting of Z
Is Eu or Ce. ), JP-A-59-56479
Europium-activated composite halogen-based phosphor BaFX.xNaX ': aEu ²⁺ (wherein X and X'are both at least one halogen selected from the group consisting of Cl, Br and I) And x is 0 <
x ≦ 2, a is 0 <a ≦ 0.2. ), JP-A-58-
MOX: xCe (here, M is Pr, Nd, Pm, Sm, Eu, Tb, Dy, which is a cerium-activated trivalent metal oxyhalide-based phosphor disclosed in Japanese Patent Publication No. 69281).
At least one trivalent metal element selected from the group consisting of Ho, Er, Tm, Yb and Bi, X is Br and I
One or both of these, x is 0 <x <0.1. ), JP-A-60-101179 and 60.
LnOX: xCe, which is a cerium-activated rare earth oxyhalogen-based phosphor disclosed in Japanese Patent Publication No. 90288/90 (wherein Ln is at least one rare earth element selected from the group consisting of Y, La, Gd and Lu, and X is At least one halogen selected from the group consisting of Cl, Br and I, x is 0 <x ≦ 0.1) and JP-A-59-59.
Europium activated complex halide-based items has been disclosed in Japanese -75200 phosphor ^{M II FX · aM I X '} · bM'
^II X ″ ₂ · cM ^III X ′ ″ ₃ · xA: yEu ²⁺ (Here,
M ^II is at least one alkaline earth metal element selected from the group consisting of Ba, Sr and Ca, and M ^I is Li, N
at least one alkali metal element selected from the group consisting of a, K, Rb and Cs, M ′ ^II is Be and Mg
At least one divalent metal element selected from the group consisting of, M ^III is at least one trivalent metal element selected from the group consisting of Al, Ga, In and Tl, A is at least one metal oxide, and X is Cl. , At least one halogen selected from the group consisting of Br and I, X ′,
X ″ and X ′ ″ are at least one halogen selected from the group consisting of F, Cl, Br and I, and a is
0 ≦ a ≦ 2, b is 0 ≦ b ≦ 10 ⁻² , c is 0 ≦ c ≦ 1
0 ^-2 and, a + b + c ≧ 10 - is ^2, x is 0
<X ≦ 0.5 and y is 0 <y ≦ 0.2. )But,
It can be preferably used.

FIG. 13 is a perspective view showing a radiation-accumulating sheet, and FIG. 13A is a layer 3 made of the stimulable phosphor described above.
4 is formed on the entire surface of the support 35, and (b)
Is the same as the arrangement pattern of the holes 3 provided in the substrate 2 of the biochemical analysis unit, and the layer 34 made of a stimulable phosphor is arranged on the support 35.

The probe molecule immobilized on the pore surface of the porous material of the biochemical analysis unit of the present invention and the target molecule labeled with the radioactive labeling substance are specifically bound to each other, and then the accumulative sheet is obtained. Is superposed on the biochemical analysis unit to accumulate radiation from the radiolabeled substance in the stimulable phosphor layer 34 of the stimulable sheet (hereinafter referred to as exposure).

A method of detecting the signal of the radiolabeled substance recorded in the stimulable phosphor layer 34 of the stimulable sheet (hereinafter referred to as an image) is shown below, but the method is not necessarily limited to this.

FIG. 14 is a schematic perspective view showing an example of an image reading apparatus for reading an image of the radioactive labeling substance recorded on the stimulable phosphor layer 34 to generate image data.
This image reading apparatus includes a first laser excitation light source 41 that emits a laser light 44 having a wavelength of 640 nm and a 532 nm laser light.
Second laser excitation light source 42 that emits laser light of the wavelength
And a third laser excitation light source 43 which emits laser light having a wavelength of 473 nm. In this embodiment,
The first laser pumping light source 41 is composed of a semiconductor laser light source, and the second laser pumping light source 42 and the third laser pumping light source 43 are second harmonic generation (Second Harmoni).
c Generation) element.

The laser light 44 generated by the first laser excitation light source 41 is collimated by the collimator lens 45 and then reflected by the mirror 46. The laser beam 44 emitted from the first laser excitation light source 41 and reflected by the mirror 46 has a first dichroic mirror 47 that transmits a laser beam of 640 nm and reflects a light of a wavelength of 532 nm and an optical path of 532 nm or more. The second dichroic mirror 48 which transmits the light of the wavelength of 473 nm and reflects the light of the wavelength of 473 nm is provided, and the laser light 44 generated by the first laser excitation light source 41 is the first laser light 44.
The light passes through the dichroic mirror 47 and the second dichroic mirror 48 and enters the optical head 55.

On the other hand, the laser light generated by the second laser excitation light source 42 is collimated by the collimator lens 50, reflected by the first dichroic mirror 47, and its direction is changed by 90 degrees. , Passes through the second dichroic mirror 48 and enters the optical head 55. Further, the laser light generated from the third laser excitation light source 43 is collimated by the collimator lens 51, reflected by the second dichroic mirror 48, and its direction is changed by 90 degrees. Head 55
Incident on.

The optical head 55 includes a mirror 56, a perforated mirror 58 having a hole 57 formed in the center thereof, and a convex lens 59. The laser light 44 incident on the optical head 55 is reflected by the mirror 56. The reflected light passes through the hole 57 formed in the perforated mirror 58 and the convex lens 59, and enters the photostimulable phosphor layer 34 carrying the fluorescent image placed on the glass plate 61 of the stage 60. . Figure 1
In No. 4, the stimulable phosphor layer 34 is placed on the glass plate 61 of the stage 60 so as to face downward.

The fluorescent light 65 emitted from the photostimulable phosphor layer 34 is collimated by the convex lens 59, reflected by the perforated mirror 58, and incident on the concave mirror 66. The fluorescence 65 that has entered the concave mirror 66 is condensed on the concave mirror 67. The fluorescent light 65 focused on the concave mirror 67 is reflected downward by the concave mirror 67, enters the filter unit 68, cuts light of a predetermined wavelength, enters the photomultiplier 69, and is detected photoelectrically. To be done. The analog image data generated photoelectrically detected by the photomultiplier 69 is
The digital image data is converted by the A / D converter 70 and sent to the image data processing device 71. Although not shown in FIG. 14, the optical head 55 is configured to be movable by the scanning mechanism so that the entire plane of the stimulable phosphor layer 34 can be scanned, and the stimulable phosphor layer 3 is provided.
The entire surface of 4 is scanned by the laser beam 44.

[0080]

EXAMPLES Example 1 (1) Fabrication of Substrate Having Through Holes A polyester sheet (substrate material sheet) having a size of 80 mm × 80 mm and a thickness of 120 μm, and a fine hole having a circular opening with a hole diameter of 0.2 mm. With a hole pitch of 0.3 mm and a hole interval of 0.1 mm, with a total of 6400 with 10 × 10 as one unit
Individually formed. The average density of the substrate material sheet is 1.0 g / c
It was m ³ .

[0081] (2) Formation of porous structure       Polysulfone (P-3500, UCC) 15 parts       72 parts of N-methyl-2-pyrrolidone       Polyvinylpyrrolidone 13 parts       1.2 parts of water

The above materials were uniformly dissolved to prepare a solution of a material for forming a porous structure. In a stable solution state, this solution was cast on a substrate material sheet by a casting coater so as to have a dry film thickness of 120 μm, and the solution was injected into the through holes, and then a surplus of the surface of the substrate material sheet was applied with a blade. The neat solution was scraped off. Next, air adjusted to a temperature of 25 ° C. and a relative humidity of 50% was sent to the surface of the liquid film of the substrate material sheet at a wind speed of 1.2 m / sec, and immediately thereafter, the substrate material sheet was immediately filled with water at 25 ° C. And a large number of fine holes were formed in the film in the through holes. Then 75
It was treated in diethylene glycol at 5 ° C for 5 minutes, then washed with water and dried to obtain a unit for biochemical analysis consisting of a polyester partition wall and a porous polymer-filled region. The average pore size of the micropores of the porous polymer was 0.2 μm.

Example 2 (1) Fabrication of substrate having through holes 80 mm × 80 mm in size, 100 μm thick SUS
Micro-holes having a circular opening with a diameter of 0.2 mm are formed in 304 sheets (substrate material sheet) by etching, with a hole pitch of 0.3 mm and a hole interval of 0.1 mm, and a total of 6400 pieces are formed as 10 × 10 pieces as one unit. did.

[0084] (2) Formation of porous structure       Nylon 6 (manufactured by Polysciences) 14 parts       66 parts formic acid       20 parts of water

The above materials were uniformly dissolved to prepare a solution of a material for forming a porous structure. In a stable solution state, this solution was cast on a substrate material sheet by a casting coater so that the thickness of the dried film was 160 μm, and the solution was injected into the through holes. The neat solution was scraped off. Then, immediately, the substrate material sheet was dipped in a coagulation bath filled with a 40% formic acid aqueous solution to form a large number of fine holes in the film in the through holes. Then, it was washed with water and dried to obtain a biochemical analysis unit consisting of a stainless partition wall and a polymer-filled region with a large number of holes.
The average pore size of the micropores of the porous polymer was 0.5 μm.

[0086] (3) Formation of porous structure       Nylon 6 (manufactured by Polysciences) 14 parts       66 parts formic acid       20 parts of water

The above materials were uniformly dissolved to prepare a solution of a material for forming a porous structure. This solution was cast on a polyester base sheet with a casting coater in a stable solution state so that the thickness of the dried film was 160 μm. Next, this substrate sheet was immediately immersed in a coagulation tank filled with a 40% formic acid aqueous solution to form a large number of fine pores in the film. Then, it was washed with water and dried, and the film was peeled off from the polyester base sheet to obtain a porous structure. The average pore size of the micropores of the porous polymer was 0.5 μm.

(4) Formation of biochemical analysis unit The porous structure obtained in (3) and the substrate obtained in (1) were superposed and fed between a press roll heated to 150 ° C. and a backup roll. , A pressure of 20 kgf / cm ² was applied to press the porous structure into the holes of the substrate to obtain a biochemical analysis unit.

(5) Evaluation of biochemical analysis unit A single-stranded nucleic acid fragment (probe molecule) was spotted on the porous structure region of the biochemical analysis unit obtained in Examples 1 and 2 by a conventional method. After fixing by adhesion, this biochemical analysis unit was immersed in an aqueous solution of a sample in which a single-stranded nucleic acid fragment (target molecule) complementary to the probe molecule was radioactively labeled to carry out hybridization. The biochemical analysis unit was taken out of the aqueous solution, washed with water, and dried. Next, it was superposed on the stimulable phosphor sheet and autoradiographed at room temperature. When a radiation image of this stimulable phosphor sheet was read using a radiation image information reading device, it was found that the porous structure region of the biochemical analysis unit (the radiolabeled target molecule was bound and fixed to the probe molecule by hybridization). The radiographic image of the area) was obtained with high resolution and high sensitivity.

[0090]

According to the method of the present invention, a plurality of holes in a substrate are filled with an adsorptive material to form the adsorptive regions, so that a unit for biochemical analysis can be efficiently prepared. be able to. By this, the specific binding substance,
When detecting radiation or light emitted by binding to a substance that is of biological origin and is labeled, and performing biochemical analysis, noise due to scattering of radiation or light is prevented, and analysis accuracy is improved. A biochemical analysis unit that can improve the

When the biochemical analysis unit produced according to the method of the present invention is used for the analysis of a bio-related substance utilizing a biochemical specific binding reaction, a probe molecule such as a nucleic acid fragment is added to the porous structure region. It can be selectively fixed on the spot. As a result, it is possible to prevent the diffusion of the probe molecule when the probe molecule solution is spotted, and to prevent the diffusion of the radiolabeled target molecule and the non-specific adhesion which are likely to occur during the analysis work. As a result, noise in the radiation image obtained by autoradiography can be reduced, and a radiation image with high resolution can be obtained. In addition, the partition walls can prevent the scattering of the radiation from the target molecules, which also improves the resolution of the radiation image. Therefore, the accuracy of detection and analysis of biological substances can be significantly improved. further,
The biochemical analysis unit according to the present invention can increase the number of absorptive regions corresponding to conventional spots per unit area, and can achieve higher density than conventional porous sheets.

[Brief description of drawings]

FIG. 1 is a perspective view showing an outline of a biochemical analysis unit according to the present invention.

FIG. 2 is a perspective view showing a method of forming a perforated substrate by punching using a plurality of pins.

FIG. 3 is a perspective view showing a method for producing a perforated substrate by photolithography and etching.

FIG. 4 is a schematic diagram showing a method of continuously injecting a dope into a perforated substrate to form an absorptive region.

FIG. 5 is a schematic view showing a method of intermittently injecting a dope into a perforated substrate.

FIG. 6 is a schematic view showing a method of binding a coupling agent to the surface of holes in a metal substrate.

FIG. 7 is a schematic view showing a method of bonding a coupling agent to the surface of holes of a plastic substrate.

FIG. 8 is a schematic view showing a method of press-fitting a porous film into the holes of a perforated substrate.

9 is an enlarged cross-sectional view taken along the line IX-IX in FIG.

FIG. 10 is a plan view showing an example of a variation of the arrangement of the absorptive regions.

FIG. 11 is a plan view showing an example of another shape of the opening of the absorptive region.

FIG. 12 is a plan view showing an example of another shape of the opening of the absorptive region.

FIG. 13 is a perspective view showing the outline of the configuration of a radiation storage sheet.

FIG. 14 is a schematic diagram showing an example of an image reading apparatus that reads a signal of a radiolabeled substance recorded in a stimulable phosphor layer to generate image data.

[Explanation of symbols]

1 Biochemical analysis unit 2 substrates 3 holes 4 Porous material (adsorptive area) 5 Specific binding substances 9 Punching pin 10 Support 11 casting die 12 blades 13 Coagulation bath 14 water bath 31 metal oxide layer 32 Coupling agent 33 metal oxide particles 34 Photostimulable phosphor layer 35 Storage sheet support

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) G01N 33/543 501 G01N 33/58 A 33/58 Z 37/00 102 37/00 102 C12M 1/00 A // C12M 1/00 C12Q 1/68 A C12N 15/09 G01N 1/28 J C12Q 1/68 C12N 15/00 F (72) Inventor Masahiro Eto 2-26-30, Nishiazabu, Minato-ku, Tokyo Shin Makoto Fuji F-term (reference) in film Co., Ltd. FD20 GA11 GA18 GA30 HB06 HC27 HC34 JA07 JA09 JA11 JA13 JA14 JA15 JA16 4B024 AA11 CA01 CA11 HA12 HA14 4B029 AA07 AA23 FA12 GA03 4B063 QA01 QA05 QA18 QR54 QS03 QS 15 QS34 QS36 QS39 QX02 QX07

Claims (17)

[Claims] 1. A biochemistry comprising a substrate formed of a material having a property of attenuating radiation and / or light and having a plurality of holes formed therein, and an absorptive region being formed in each of the plurality of holes. In the method for manufacturing an analysis unit, a method for manufacturing a biochemical analysis unit, characterized in that an adsorptive material is filled in each of the plurality of holes to form the adsorptive region. 2. Formed by a material having a property of attenuating radiation and / or light, a plurality of holes are formed, and an absorptive region is formed in each of the plurality of holes, whereby a plurality of absorptive regions are formed. Of the substrate, the specific binding substance having a known structure or characteristic is dripped into the plurality of absorptive regions formed in the plurality of holes of the substrate, the radiolabeled substance, the fluorescent substance. A substance derived from a living body, which is labeled with at least one labeling substance selected from the group consisting of labeling substances that generate chemiluminescence when brought into contact with either a substance or a chemiluminescent substrate, specifically binds to the specific binding substance. In the method of manufacturing a unit for biochemical analysis, which is bound, the plurality of absorptive regions are selectively labeled, in each of the plurality of holes, an absorptive material is provided Hama to method for producing a biochemical analysis unit, characterized in that to form the absorptive regions. 3. A plurality of absorptive regions formed by forming an absorptive region in each of the plurality of holes of a substrate formed of a material having a property of attenuating radiation and having a plurality of holes formed therein. In the region, a specific binding substance capable of specifically binding to a biological substance and having a known structure or property is dropped, and the biological substance labeled with a radioactive labeling substance is added to the specific binding substance. By specifically binding, the plurality of adsorptive regions are selectively labeled, and the stimulable phosphor layer is adsorbed by the stimulable phosphor layer on the stimulable phosphor sheet on which the stimulable phosphor layer is formed. To face the sex area,
Overlaid, by the radioactive labeling substance contained in the plurality of adsorptive regions, the stimulable phosphor layer is exposed.
Irradiating the stimulable phosphor layer exposed by the radioactive labeling substance with excitation light to excite the stimulable phosphor contained in the stimulable phosphor layer, and the stimulable phosphor layer. Photosensitized photostimulable light emitted from the stimulable phosphor contained in, in the method of manufacturing a biochemical analysis unit for generating biochemical analysis data, in the plurality of holes A method for manufacturing a biochemical analysis unit, characterized in that each of them is filled with an adsorptive material to form the adsorptive region. 4. The adsorbent material solution, which comprises a good solvent for the adsorbent material as a main component of the solvent composition of the adsorbent material and a small amount of a poor solvent, is injected into the plurality of holes of the substrate, and then the substrate is removed. The method for producing a biochemical analysis unit according to claim 1, wherein the adsorptive material is filled by being dipped in a coagulation liquid or gradually dried. 5. The absorptive material formed in advance in the form of a film and the substrate are overlapped and pressed to press the film-like absorptive material into the holes of the substrate to obtain the absorptive property. The method for manufacturing a biochemical analysis unit according to any one of claims 1 to 3, wherein the material is filled. 6. The method for manufacturing a biochemical analysis unit according to claim 1, wherein the adsorptive material is a porous material. 7. The porous material comprises a material capable of forming a membrane filter.
A method for manufacturing the biochemical analysis unit described in. 8. The method for manufacturing a biochemical analysis unit according to claim 6, wherein a fibrous material insoluble in the solvent of the porous material is mixed with the porous material. 9. Radiation and / or light when the material having a property of attenuating the radiation and / or light is transmitted through the material by a distance equal to the distance between adjacent absorptive regions. The method for manufacturing a biochemical analysis unit according to any one of claims 1 to 8, wherein the method has a property of attenuating the energy of light and / or light to 1/5 or less. 10. The substrate according to claim 1, wherein the substrate is made of a material selected from the group consisting of a metal material, a ceramic material and a plastic material.
A method for manufacturing the biochemical analysis unit according to any one of 1. 11. The method for manufacturing a biochemical analysis unit according to claim 1, wherein a method of forming the plurality of holes in the substrate is a punching method. 12. The method of manufacturing a biochemical analysis unit according to claim 1, wherein a method of forming the plurality of holes in the substrate is an electric discharge machining method. 13. The biochemical analysis according to claim 1, wherein the method of forming the plurality of holes in the substrate is a method of combining photolithography and etching. Method for manufacturing units. 14. The method for manufacturing a biochemical analysis unit according to claim 1, wherein a method of forming the plurality of holes in the substrate is a laser ablation method. 15. The method for manufacturing a biochemical analysis unit according to claim 1, wherein the size of the hole is less than 5 mm ² . 16. The method for manufacturing a biochemical analysis unit according to claim 1, wherein the holes are formed at a density of 10 holes / cm ² or more. 17. A biochemical analysis unit manufactured by the method according to any one of claims 1 to 16.