JP3920054B2 - Method and apparatus for generating data for biochemical analysis - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method and apparatus for generating data for biochemical analysis, and more specifically, the present invention is capable of specifically binding to a substance derived from a living body, and has a base sequence and a base length, A specific binding substance whose composition is known is dropped onto the carrier in a spot shape to form a spot-like region at high density, and the specific binding substance contained in the spot-like region is brought into contact with the chemiluminescent substrate. Chemiluminescence emitted from a unit for biochemical analysis obtained by specifically binding and selectively labeling a biological substance labeled with a labeling substance and / or a fluorescent substance that generates chemiluminescence Even when fluorescence is detected photoelectrically, biochemical analysis data is generated, and a biological substance is analyzed, noise due to chemiluminescence and / or fluorescence scattering is also detected in the data for biochemical analysis. Relates generating method and apparatus of the biochemical analysis data can be prevented from being generated during.
[0002]
[Prior art]
When irradiated with radiation, the energy of the radiation is absorbed, stored, recorded, and then excited using electromagnetic waves in a specific wavelength range. After using a stimulable phosphor having the property of emitting light as a radiation detection material, a radioactively labeled substance is administered to the organism, and then the organism or a part of the tissue of the organism is used as a sample. The sample is superposed on a stimulable phosphor sheet provided with a stimulable phosphor layer for a certain period of time, whereby radiation energy is accumulated and recorded in the stimulable phosphor, and then the sample is illuminated by electromagnetic waves. The stimulable phosphor layer is scanned to excite the stimulable phosphor, and the photostimulated light emitted from the stimulable phosphor is photoelectrically detected to generate a digital image signal and perform image processing. On the display means such as CRT Or, an autoradiography analysis system configured to reproduce an image on a recording material such as a photographic film is known (for example, Japanese Patent Publication No. 1-70884, Japanese Patent Publication No. 1-70882, No. 4-3962).
[0003]
Unlike radiographic systems, autoradiographic analysis systems that use stimulable phosphor sheets as radiation detection materials do not require chemical processing, such as development processing, but can also be applied to the resulting digital data. By performing the data processing, there is an advantage that the data for analysis can be reproduced as desired or quantitative analysis by a computer can be performed.
[0004]
On the other hand, a fluorescence analysis system using a fluorescent substance such as a fluorescent dye as a labeling substance instead of the radioactive labeling substance in the autoradiography analysis system is known. According to this fluorescence analysis system, by detecting the fluorescence emitted from the fluorescent substance, gene sequence, gene expression level, metabolism, absorption, excretion route, state of the administered substance in the experimental mouse, separation of proteins, Identification, evaluation of molecular weight, characteristics, etc. can be performed. For example, after electrophoresis of a solution containing multiple types of protein molecules to be electrophoresed on a gel support, the gel support is fluorescent. Stain the electrophoretic protein by soaking in a solution containing the dye, etc., excite the fluorescent dye with excitation light, and detect the resulting fluorescence to generate an image on the gel support The position and quantitative distribution of protein molecules can be detected. Alternatively, at least a part of the electrophoresed protein molecule is transferred onto a transfer support such as nitrocellulose by Western blotting, and an antibody that specifically reacts with the target protein is labeled with a fluorescent dye. By associating the prepared probe with the protein molecule, selectively labeling the protein molecule that binds only to the antibody that reacts specifically, exciting the fluorescent dye with excitation light, and detecting the resulting fluorescence, An image 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. Electrophoresis of a plurality of DNA fragments, or electrophoresis of a plurality of DNA fragments on a gel support followed by immersing the gel support in a solution containing a fluorescent dye. By labeling the DNA fragments, exciting the fluorescent dye with excitation light, and detecting the resulting fluorescence, an image is generated and the distribution of DNA on the gel support is detected, or a plurality of DNAs are detected. After the fragments are electrophoresed on a gel support, the DNA is denaturated, and then at least the denatured DNA fragments are transferred onto a transfer support such as nitrocellulose by Southern blotting. In addition, a probe prepared by transferring a part of the DNA and labeling the DNA or RNA complementary to the target DNA with a fluorescent dye is hybridized with the denatured DNA fragment, and only the DNA fragment complementary to the probe DNA or probe RNA. Can be selectively labeled, the fluorescent dye can be excited with excitation light, and the resulting fluorescence can be detected to generate an image and detect the distribution of the target DNA on the transfer support. it can. Further, a DNA probe complementary to the DNA containing the target gene labeled with the labeling substance is prepared, hybridized with the DNA on the transcription support, and the enzyme is combined with the complementary DNA labeled with the labeling substance. After binding, contact the fluorescent substrate, change the fluorescent substrate into a fluorescent substance that emits fluorescence, excite the generated fluorescent substance with excitation light, and generate the image by detecting the generated fluorescence It is also possible to detect the distribution of the target DNA on the transfer support. This fluorescence analysis system has an advantage that gene sequences and the like can be easily detected without using a radioactive substance.
[0005]
Similarly, biologically-derived substances such as proteins and nucleic acids are immobilized on a support, selectively labeled with a labeling substance that generates chemiluminescence by contacting with a chemiluminescent substrate, and selectively selected with a labeling substance. A chemiluminescent substrate in contact with a labeled biological substance and a chemiluminescent substrate is contacted, and the chemiluminescence in the visible light wavelength region generated by the contact between the chemiluminescent substrate and the labeling substance is photoelectrically detected to generate a digital image signal. There is also a chemiluminescence analysis system that performs image processing and reproduces a chemiluminescence image on a display means such as a CRT or a recording material such as a photographic film to obtain information on a substance derived from a living body such as genetic information. Are known.
[0006]
Furthermore, in recent years, hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA, etc. A specific binding substance that can specifically bind to the substance and has a known base sequence, base length, composition, etc. is dropped using a spotter device to form a large number of independent spots, Subsequently, hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, mRNA, etc. are collected from the living body by extraction, isolation, etc., or further, chemical treatment, chemistry A substance derived from a living body that has been subjected to treatment such as modification, and that is labeled with a labeling substance such as a fluorescent substance or a dye is hybridized. By irradiating excitation light to a microarray that is specifically bound to a specific binding substance by means of, for example, fluorescence, light such as fluorescence emitted from a labeling substance such as a fluorescent substance or a dye is detected photoelectrically, Microarray analysis systems that analyze biological materials have been developed. According to this microarray analysis system, a large number of spots of specific binding substances are formed in high density at different positions on the surface of a carrier such as a slide glass plate or a membrane filter, and a biological substance labeled with a labeling substance. By hybridizing, there is an advantage that it becomes possible to analyze a substance derived from a living body in a short time.
[0007]
In addition, it specifically binds to biological substances such as hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA at different positions on the carrier surface such as membrane filters. A specific binding substance having a known base sequence, base length, composition, etc., is dropped using a spotter device to form a large number of independent spots, and then hormones, tumors Markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, mRNA, etc. are collected from living bodies by extraction, isolation, etc., or further subjected to chemical treatment, chemical modification, etc. A biologically-derived substance that has been labeled with a radioactive labeling substance, a specific binding substance by hybridization or the like In addition, the specifically bonded macroarray is brought into close contact with the stimulable phosphor sheet on which the photostimulable phosphor layer containing the photostimulable phosphor is formed, and the photostimulable phosphor layer is exposed. Later, the stimulable phosphor layer is irradiated with excitation light, and the photostimulated light emitted from the stimulable phosphor layer is detected photoelectrically to generate biochemical analysis data. A macroarray analysis system using a radiolabeled substance to be analyzed has also been developed.
[0008]
[Problems to be solved by the invention]
In recent years, it specifically binds to biological substances such as hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA, etc. at different positions on the surface of the carrier such as membrane filters. A specific binding substance that is possible and has a known base sequence, base length, composition, etc. is dropped to form a plurality of spot-like regions, and the specific binding substances contained in the plurality of spot-like regions are formed. A biologically-labeled substance labeled with a labeling substance and / or a fluorescent substance that generates chemiluminescence by contacting with a chemiluminescent substrate is specifically bound by hybridization or the like, and selectively labeled. The chemiluminescence produced by the contact between the chemiluminescent substrate and the labeling substance is photoelectrically detected and / or the excitation light is contacted with the luminescent substrate. However, it is also required that the fluorescence emitted from the fluorescent material is photoelectrically detected by a photodetector such as a CCD camera, data for biochemical analysis is generated, and biochemical analysis is performed. When chemiluminescence or fluorescence is photoelectrically detected by a photodetector, chemiluminescence or fluorescence emitted from the spot-like region is scattered within the carrier, or chemiluminescence or fluorescence emitted from the spot-like region. Are scattered and mixed with chemiluminescence and fluorescence emitted from adjacent spot-like regions, and as a result, noise is generated in biochemical analysis data generated by photoelectrically detecting chemiluminescence and / or fluorescence There was a problem to do.
[0009]
Therefore, the present invention provides a specific binding substance that can specifically bind to a substance derived from a living body and has a known base sequence, base length, composition, and the like by dropping it onto a carrier in a spot shape. In the density, a spot-like region is formed, and a specific binding substance contained in the spot-like region is brought into contact with a chemiluminescent substrate to cause chemiluminescence, and is derived from a biological substance labeled with a labeling substance and / or a fluorescent substance Biochemical analysis data is generated by photoelectrically detecting chemiluminescence and / or fluorescence emitted from a biochemical analysis unit obtained by specifically binding and selectively labeling substances Even when analyzing substances of this type, it is possible to effectively prevent noise caused by chemiluminescence and / or fluorescence scattering from being generated in the data for biochemical analysis. data It is an object to provide a production method and apparatus biochemical analysis data that can be generated.
[0010]
[Means for Solving the Problems]
This object of the present invention is to A plurality of holes are two-dimensionally formed by being spaced apart from each other and two-dimensionally formed by filling the plurality of holes in the substrate with an adsorbent material and being separated from each other. Multiple adsorptive regions A biochemical analysis unit comprising: a sample stage; and light emitted from the plurality of adsorptive regions of the biochemical analysis unit placed on the sample stage for the biochemical analysis A plurality of condensing lenses arranged opposite to each of the plurality of adsorptive regions of the unit are focused on a two-dimensional solid sensor, and photoelectrically detected by the two-dimensional solid sensor, and biochemical This is achieved by a biochemical analysis data generation method characterized by generating analysis data.
[0011]
According to the present invention, the biochemical analysis unit comprises: A plurality of holes spaced apart from each other and two-dimensionally formed, and a plurality of holes formed in the substrate are filled with an adsorbent material and spaced apart from each other to form a plurality of holes formed two-dimensionally. Adsorbent area Therefore, a specific binding substance that can specifically bind to a substance derived from a living body and has a known base sequence, base length, composition, and the like is used as a plurality of absorptive regions of the biochemical analysis unit. A specific substance bound to a specific binding substance contained in a plurality of adsorptive regions is specifically bound to a biological substance labeled with a labeling substance that generates chemiluminescence by contacting with the chemiluminescent substrate, When chemiluminescence emitted from the biochemical analysis unit obtained by selective labeling is photoelectrically detected to generate biochemical analysis data, chemiluminescence emitted from adjacent adsorbent regions is mixed. It is possible to effectively prevent noise from being generated in biochemical analysis data due to the scattering of chemiluminescence. Excellent raw It is possible to generate data for academic analysis.
[0012]
Further, according to the present invention, the biochemical analysis unit comprises: A plurality of holes spaced apart from each other and two-dimensionally formed, and a plurality of holes formed in the substrate are filled with an adsorbent material and spaced apart from each other to form a plurality of holes formed two-dimensionally. Adsorbent area Therefore, a specific binding substance that can specifically bind to a substance derived from a living body and has a known base sequence, base length, composition, and the like is used as a plurality of absorptive regions of the biochemical analysis unit. A biologically-derived substance labeled with a fluorescent substance is specifically bound to a specific binding substance contained in a plurality of adsorptive regions, selectively labeled, and a plurality of biochemical analysis units By irradiating the adsorbing region with excitation light, the fluorescent material selectively contained in the plurality of adsorbing regions is excited, and the fluorescence emitted from the fluorescent material is photoelectrically passed through the excitation light cut filter. When detecting and generating data for biochemical analysis, it is possible to effectively prevent the fluorescence emitted from the adjacent adsorptive regions from being mixed, and thus noise due to the scattering of the fluorescence is generated. Generated in data for chemical analysis Since it becomes possible to effectively prevent, it is possible to produce biochemical analysis data having an excellent quantitative characteristic.
[0013]
Furthermore, according to the present invention, the light emitted from the plurality of absorptive regions of the biochemical analysis unit placed on the sample stage is opposed to each of the plurality of absorptive regions of the biochemical analysis unit. The two-dimensional solid-state sensor condenses light on the two-dimensional solid-state sensor using a plurality of arranged condensing lenses, and is photoelectrically detected by the two-dimensional solid-state sensor to generate biochemical analysis data. Light emitted from the plurality of absorptive regions with a plurality of condensing lenses facing each of the plurality of absorptive regions of the chemical analysis unit sufficiently close to the plurality of absorptive regions of the biochemical analysis unit By collecting the light, it is possible to collect the light emitted from the plurality of adsorptive regions with high light collection efficiency and guide it to the two-dimensional solid state sensor, and to emit from each of the plurality of adsorptive regions. Light to be The angle at which the condensing lens is viewed from each adsorptive region becomes smaller and the stray light is not likely to be generated in the lens as in the case of guiding to the two-dimensional solid sensor by one condensing lens. The two-dimensional solid-state sensor can detect light emitted from a plurality of adsorptive regions with high sensitivity and generate biochemical analysis data with high quantitativeness.
[0014]
In a preferred embodiment of the present invention, the substrate of the biochemical analysis unit has a property of attenuating light.
[0015]
According to a preferred embodiment of the present invention, the substrate of the biochemical analysis unit has the property of attenuating light. Formed by filling adsorbent material in multiple holes Effectively prevent chemiluminescence or fluorescence emitted from each adsorptive region from being scattered within the substrate of the biochemical analysis unit and mixing chemiluminescence or fluorescence emitted from adjacent adsorbent regions. Therefore, it is possible to effectively prevent noise caused by chemiluminescence or fluorescence scattering from being generated in the data for biochemical analysis. Can be generated.
[0016]
In a preferred embodiment of the present invention, when the substrate of the biochemical analysis unit transmits light through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is reduced to 1 /. It has the property of being attenuated to 5 or less.
[0017]
In a further preferred embodiment of the present invention, when the light of the substrate of the biochemical analysis unit is transmitted through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is set to 1. / 10 or less.
[0018]
In a further preferred embodiment of the present invention, when the light of the substrate of the biochemical analysis unit is transmitted through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is set to 1. / 50 or less.
[0019]
In a further preferred embodiment of the present invention, when the light of the substrate of the biochemical analysis unit is transmitted through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is set to 1. / 100 or less.
[0020]
In a further preferred embodiment of the present invention, when the light of the substrate of the biochemical analysis unit is transmitted through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is set to 1. / 500 or less.
[0021]
In a further preferred embodiment of the present invention, when the substrate of the biochemical analysis unit transmits light through the substrate by a distance equal to the distance between adjacent adsorptive regions, the light energy is It has the property of being attenuated to 1/1000 or less.
[0022]
In a further preferred embodiment of the present invention, the condenser lens is constituted by a lens having a large numerical aperture.
[0023]
In a further preferred embodiment of the present invention, the plurality of condensing lenses are arranged in the lens array so that the pitch of the adjacent condensing lenses is equal to the pitch of the adjacent adsorptive regions of the biochemical analysis unit. It is attached.
[0024]
In a preferred embodiment of the present invention, the two-dimensional solid sensor is constituted by a cooled CCD sensor.
[0025]
According to a preferred embodiment of the present invention, since the two-dimensional solid-state sensor is constituted by a cooled CCD sensor, light emitted from a plurality of adsorptive regions can be detected over a long period of time. It is possible to detect chemiluminescence or fluorescence, which is a simple light, with high sensitivity and to generate biochemical analysis data with excellent quantification.
[0026]
In a preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit includes a specific binding substance, and the specific binding substance contained in the plurality of absorptive regions is chemically treated. A biologically-derived substance labeled with a labeling substance that generates chemiluminescence by being brought into contact with a luminescent substrate is selectively and specifically bound to chemiluminescence emitted from the plurality of adsorptive regions. A three-dimensional solid-state sensor is configured to photoelectrically detect and generate biochemical analysis data.
[0027]
In a preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit contains a specific binding substance, and the specific binding substances contained in the plurality of absorptive regions are fluorescent. A biological substance labeled with a substance is selectively and specifically bound to the plurality of absorptive regions of the biochemical analysis unit, opposite to the two-dimensional solid sensor with respect to the sample stage. Excitation light emitted from an excitation light source located on the side is irradiated to excite the fluorescent material selectively contained in the plurality of adsorptive regions, and the fluorescence emitted from the fluorescent material is collected into the plurality of light collections. The lens cuts the light having the wavelength of the excitation light and guides it to the two-dimensional sensor through an excitation hole cut filter having a property of transmitting light having a wavelength longer than that of the excitation light. Accordingly, by photoelectrically detecting it is configured to produce biochemical analysis data.
[0028]
In the present invention, the labeling with the fluorescent substance means that the fluorescent substance is labeled with the fluorescent dye, and the enzyme is contacted with the fluorescent substrate after binding the enzyme with the labeled sample, And a case where the fluorescent substance is labeled with the obtained fluorescent substance.
[0029]
In a preferred embodiment of the present invention, the biological substance is bound to the specific binding substance by a reaction selected from the group consisting of hybridization, antigen-antibody reaction, and receptor / ligand.
[0030]
In a preferred embodiment of the present invention, 10 or more absorptive regions are formed on the substrate of the biochemical analysis unit.
[0031]
In a further preferred embodiment of the present invention, 50 or more absorptive regions are formed on the substrate of the biochemical analysis unit.
[0032]
In a further preferred embodiment of the present invention, 100 or more absorptive regions are formed on the substrate of the biochemical analysis unit.
[0033]
In a further preferred embodiment of the present invention, 500 or more absorptive regions are formed on the substrate of the biochemical analysis unit.
[0034]
In a further preferred embodiment of the present invention, 1000 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.
[0035]
In a further preferred embodiment of the present invention, 5000 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.
[0036]
In a further preferred embodiment of the present invention, 10,000 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.
[0037]
In a further preferred embodiment of the present invention, 50000 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.
[0038]
In a further preferred embodiment of the present invention, 100,000 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.
[0039]
In a preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 5 square millimeters.
[0040]
In a further preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 1 square millimeter.
[0041]
In a further preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 0.5 square millimeters.
[0042]
In a further preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 0.1 mm 2.
[0043]
In a further preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 0.05 square millimeters.
[0044]
In a further preferred embodiment of the present invention, each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 0.01 square millimeters.
[0045]
In a preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 10 / cm 2 or more.
[0046]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 50 or more per square centimeter.
[0047]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 100 or more per square centimeter.
[0048]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 500 or more per square centimeter.
[0049]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 1000 / cm 2 or more.
[0050]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 5000 or more per square centimeter.
[0051]
In a further preferred embodiment of the present invention, the plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 10,000 or more per square centimeter.
[0052]
In a preferred embodiment of the invention, In the substrate of the biochemical analysis unit, a plurality of through holes are formed two-dimensionally apart from each other, The plurality of absorptive regions are the plurality of the absorptive regions. Penetration The hole is filled with an adsorbent material.
[0054]
In a further preferred embodiment of the present invention, the plurality of adsorptive regions are formed by press-fitting an adsorbing film containing an adsorbing material into the plurality of through holes of the substrate. .
[0055]
According to a preferred embodiment of the present invention, the plurality of adsorptive regions are formed by press-fitting the adsorptive film containing the adsorptive material into the plurality of through holes of the substrate. It is possible to produce biochemical analysis units with adsorptive regions .
[0056]
In another preferred embodiment of the present invention, a plurality of recesses are two-dimensionally formed on the substrate of the biochemical analysis unit so as to be spaced apart from each other, and the plurality of adsorptive regions are the plurality of the plurality of adsorbing regions. The recess is filled with an adsorbent material and formed. .
[0057]
In a preferred embodiment of the present invention, the plurality of adsorptive regions are regularly formed on the substrate of the biochemical analysis unit.
[0058]
In the present invention, the material for forming the substrate of the biochemical analysis unit preferably has the property of attenuating light, but is not particularly limited, and is not limited to inorganic compound materials and organic compound materials. Any of them can be used, and a metal material, a ceramic material or a plastic material is preferably used.
[0059]
In the present invention, as an inorganic compound material that can be preferably used to form a substrate for a biochemical analysis unit, for example, gold, silver, copper, zinc, aluminum, titanium, tantalum, chromium, iron, nickel, Metals such as cobalt, lead, tin, and selenium; alloys such as brass, stainless steel, and bronze; silicon materials such as silicon, amorphous silicon, glass, quartz, silicon carbide, and silicon nitride; metals such as aluminum oxide, magnesium oxide, and zirconium oxide Examples of oxides include inorganic salts such as tungsten carbide, calcium carbonate, calcium sulfate, hydroxyapatite, and gallium arsenide. These may have any structure in a polycrystalline sintered body such as single crystal, amorphous, or ceramic.
[0060]
In the present invention, as an organic compound material that can be used to form a substrate for a biochemical analysis unit, a polymer compound is preferably used. Examples of a polymer compound that can be preferably used include polyethylene and polypropylene. Acrylic resins such as polymethyl methacrylate and butyl acrylate / methyl methacrylate copolymer; polyacrylonitrile; polyvinyl chloride; polyvinylidene chloride; polyvinylidene fluoride; polytetrafluoroethylene; polychlorotrifluoroethylene; Polyesters such as naphthalate and polyethylene terephthalate; nylons such as nylon 6, nylon 6,6, nylon 4,10; polyimide; polysulfone; polyphenylenesulfur Silicone resin such as polydiphenylsiloxane; phenolic resin such as novolac; epoxy resin; polyurethane; polystyrene; butadiene-styrene copolymer; polysaccharide such as cellulose, cellulose acetate, nitrocellulose, starch, calcium alginate, hydroxypropylmethylcellulose, etc. Chitin; chitosan; urushi; polyamides such as gelatin, collagen, keratin, and copolymers of these polymer compounds. These may be composite materials, and if necessary, may be filled with metal oxide particles or glass fibers, or may be used by blending organic compound materials.
[0061]
In general, the greater the light scattering and / or absorption, the higher the light attenuation capability. Therefore, the substrate of the biochemical analysis unit preferably has an absorbance of 0.3 or more per 1 cm in thickness. More preferably, the absorbance per cm is 1 or more. Here, the absorbance is calculated by calculating the A / T by placing an integrating sphere immediately after the Tcm-thick plate and measuring the amount of transmitted light A at the wavelength of the probe light or emission light used for measurement with a spectrophotometer. Is required. In order to improve the light attenuation ability, a light scatterer or a light absorber can be contained in the substrate of the biochemical analysis unit. As the light scatterer, fine particles of a material different from the material forming the substrate of the biochemical analysis unit are used, and as the light absorber, a pigment or a dye is used.
[0062]
In the present invention, a porous material or a fiber material is preferably used as the absorptive material forming the absorptive region of the biochemical analysis unit. The adsorptive region can also be formed by using a porous material and a fiber material in combination.
[0063]
In the present invention, the porous material used to form the adsorptive region of the biochemical analysis unit may be either an organic material or an inorganic material, or an organic / inorganic composite.
[0064]
In the present invention, the organic porous material used for forming the adsorptive region of the biochemical analysis unit is not particularly limited, but a carbon material such as activated carbon or a material capable of forming a membrane filter is used. Are preferably used. Specifically, nylons such as nylon 6, nylon 6,6, nylon 4,10; cellulose derivatives such as nitrocellulose, cellulose acetate, cellulose butyrate acetate; collagen; alginic acid, calcium alginate, alginic acid / polylysine polyion complex, etc. Examples thereof include alginic acids; polyolefins such as polyethylene and polypropylene; polyvinyl chloride; polyvinylidene chloride; polyfluorides such as polyvinylidene fluoride and polytetrafluoride, and copolymers or composites thereof.
[0065]
In the present invention, the inorganic porous material used for forming the adsorptive region of the biochemical analysis unit is not particularly limited, but preferably, for example, platinum, gold, iron, silver, nickel Metals such as aluminum; metal oxides such as alumina, silica, titania and zeolite; metal salts such as hydroxyapatite and calcium sulfate, and composites thereof.
[0066]
In the present invention, the fiber material used for forming the adsorptive region of the biochemical analysis unit is not particularly limited, but preferably, for example, nylon 6, nylon 6,6, nylon 4, Nylon such as 10, cellulose derivatives such as nitrocellulose, cellulose acetate, and cellulose acetate butyrate.
[0067]
In the present invention, a plurality of absorptive regions of the biochemical analysis unit are subjected to oxidation treatment such as electrolytic treatment, plasma treatment and arc discharge; primer treatment using a silane coupling agent, titanium coupling agent, etc .; surfactant treatment It can also be formed by surface treatment such as. It is particularly preferable that the surface of the adsorptive region has a fractal structure.
[0068]
The object of the invention is also A plurality of holes are two-dimensionally formed by being spaced apart from each other and two-dimensionally formed by filling the plurality of holes in the substrate with an adsorbent material and being separated from each other. Multiple adsorptive regions A sample stage for mounting a biochemical analysis unit comprising: a two-dimensional solid state sensor; and facing each of the plurality of adsorptive regions of the biochemical analysis unit mounted on the sample stage This is achieved by a biochemical analysis data generation device characterized in that a lens array having a plurality of condensing lenses attached thereto is provided at the position.
[0069]
According to the present invention, the data generation device for biochemical analysis comprises: A plurality of holes spaced apart from each other and two-dimensionally formed, and a plurality of holes formed in the substrate are filled with an adsorbent material and spaced apart from each other to form a plurality of holes formed two-dimensionally. Adsorbent area A sample stage for mounting a biochemical analysis unit equipped with a two-dimensional solid state sensor, and in a position facing each of a plurality of adsorptive regions of the biochemical analysis unit mounted on the sample stage. Because it has a lens array with multiple condenser lenses attached, the lens array is placed close enough to each of the multiple absorptive regions of the biochemical analysis unit placed on the sample stage. By doing so, chemiluminescence and fluorescence emitted from a plurality of adsorptive regions can be condensed with high condensing efficiency and guided to a two-dimensional solid state sensor. The angle at which the condensing lens is viewed from each adsorptive region is reduced as in the case where the emitted chemiluminescence or fluorescence is guided to the two-dimensional solid sensor by one condensing lens Because there is no possibility that stray light is easily generated in the lens, biochemistry with high quantification is detected by detecting chemiluminescence and fluorescence emitted from multiple adsorptive regions with high sensitivity using a two-dimensional solid state sensor. Analysis data can be generated.
[0070]
In a preferred embodiment of the present invention, the biochemical analysis data generation device further includes an excitation light source that emits excitation light on the opposite side of the two-dimensional solid sensor with respect to the sample stage, and the lens array. And an excitation light cut filter having a property of cutting light having the wavelength of the excitation light and transmitting light having a wavelength longer than that of the excitation light.
[0071]
According to a preferred embodiment of the present invention, the data generation device for biochemical analysis further includes an excitation light source that emits excitation light on the side opposite to the two-dimensional solid sensor with respect to the sample stage, and a lens array. Since it has an excitation light cut filter that cuts light of the wavelength of excitation light and transmits light having a wavelength longer than that of excitation light between two-dimensional solid-state sensors, it is specifically designed for biological substances. Specific binding substances that are capable of binding and whose base sequence, base length, composition, etc. are known are dropped onto the multiple absorptive regions of the biochemical analysis unit, and the specifics contained in the multiple absorptive regions A biologically-bound substance labeled with a fluorescent substance is specifically bound to a chemical binding substance, selectively labeled, and fluorescence data is recorded and excited in multiple absorptive regions of the biochemical analysis unit. Emitted from the light source The excitation light thus irradiated is irradiated to a plurality of adsorptive regions of the biochemical analysis unit to excite the fluorescent material selectively contained in the plurality of adsorptive regions, and the fluorescence emitted from the fluorescent material is When photoelectrically detecting and generating biochemical analysis data with a three-dimensional solid state sensor, a lens array with a plurality of condensing lenses facing each of the plurality of adsorptive regions is mounted on the sample stage. The fluorescent light emitted from the plurality of absorptive regions is condensed with high condensing efficiency by being placed in close proximity to each of the plurality of absorptive regions of the placed biochemical analysis unit, In the case where the fluorescence emitted from each of the plurality of adsorptive regions is guided to the two-dimensional solid sensor by a single condenser lens, it can be guided to the two-dimensional solid sensor through the excitation light cut filter. Yo In addition, since the angle at which the condensing lens is viewed from each adsorptive region is reduced and stray light is not easily generated in the lens, the two-dimensional solid sensor is used to detect the adsorptive region from a plurality of adsorptive regions. It is possible to detect the emitted fluorescence and generate highly quantitative data for biochemical analysis.
[0072]
In a preferred embodiment of the present invention, the condenser lens is constituted by a lens having a large numerical aperture.
[0073]
In a preferred embodiment of the present invention, the plurality of condensing lenses are arranged in the lens array such that the pitch of the condensing lenses adjacent to each other is equal to the pitch of the adsorbing regions adjacent to the biochemical analysis unit. It is attached.
[0074]
In a preferred embodiment of the present invention, the two-dimensional solid sensor is constituted by a cooled CCD sensor.
[0075]
According to a preferred embodiment of the present invention, since the two-dimensional solid-state sensor is constituted by a cooled CCD sensor, light emitted from a plurality of adsorptive regions can be detected over a long period of time. It is possible to detect chemiluminescence or fluorescence, which is a simple light, with high sensitivity and to generate biochemical analysis data with excellent quantification.
[0076]
In a preferred embodiment of the present invention, the excitation light source is constituted by a laser excitation light source.
[0077]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.
[0078]
FIG. 1 is a schematic perspective view of a biochemical analysis unit used in a method for generating biochemical analysis data according to a preferred embodiment of the present invention.
[0079]
As shown in FIG. 1, the biochemical analysis unit 1 according to this embodiment includes a substrate 2 formed of aluminum and having a large number of substantially circular through-holes 3 formed at a high density. The through hole 3 is filled with nylon 6 to form a large number of dot-like adsorptive regions 4.
[0080]
Although not exactly shown in FIG. 1, in this embodiment, there are regularly through-holes 3 with a density of about 5000 holes / square centimeter, with a size of about 0.01 square millimeters of about 10,000. 2 is formed. The adsorptive region 4 is formed by filling a number of through holes 3 with nylon 6 so that the surface thereof is positioned at the same height as the surface of the substrate 2.
[0081]
FIG. 2 is a schematic front view of the spotting device.
[0082]
In the biochemical analysis, as shown in FIG. 2, in a large number of adsorbing regions 4 regularly formed in the biochemical analysis unit 1, for example, as a specific binding substance, the base sequences are known to each other. Different cDNAs are dropped using the spotting device 5.
[0083]
As shown in FIG. 2, the spotting device 5 includes an injector 6 and a CCD camera 7, and the CCD camera 7 penetrates the tip of the injector 6 and the biochemical analysis unit 1 to which a specific binding substance is to be dropped. While observing the hole 3, when the tip of the injector 6 coincides with the center of the through-hole 3 where the specific binding substance should be dropped, a plurality of cDNAs whose base sequences are different from each other from the injector 6 The specific binding substance is configured to be dropped, and it is ensured that the specific binding substance can be accurately dropped into the large number of dot-like adsorptive regions 4 of the biochemical analysis unit 1. ing.
[0084]
FIG. 3 is a schematic longitudinal sectional view of the hybridization container.
[0085]
As shown in FIG. 3, the hybridization container 8 has a rectangular cross section, and contains therein a hybridization solution 9 containing a biological substance that is a probe labeled with a labeling substance.
[0086]
When a specific binding substance such as cDNA is selectively labeled with a labeling substance that generates chemiluminescence by contacting with the chemiluminescent substrate, the labeling substance that generates chemiluminescence by contacting with the chemiluminescent substrate is used. A hybridization solution 9 containing a biologically-derived substance that is a labeled probe is prepared and accommodated in a hybridization container 8.
[0087]
On the other hand, when a specific binding substance such as cDNA is selectively labeled with a fluorescent substance such as a fluorescent dye, a hybridization solution 9 containing a biological substance that is a probe labeled with the fluorescent substance such as a fluorescent dye. Is prepared and accommodated in the hybridization container 8.
[0088]
Includes two or more biologically-derived substances among biologically-derived substances labeled with a labeling substance that generates chemiluminescence upon contact with a chemiluminescent substrate and biologically-derived substances labeled with a fluorescent substance such as a fluorescent dye A hybridization solution 9 can be prepared and accommodated in the hybridization container 8. In this embodiment, a biological substance labeled with a labeling substance that generates chemiluminescence by contacting with the chemiluminescent substrate. A hybridization solution 9 containing a biologically-derived substance labeled with a fluorescent substance is prepared and accommodated in the hybridization container 8.
[0089]
In hybridization, a biochemical analysis unit 1 in which a specific binding substance such as cDNA is adsorbed in a number of adsorptive regions 4 is inserted into the hybridization container 8.
[0090]
As a result, a biologically-derived substance labeled with a labeling substance that causes chemiluminescence to be brought into contact with a specific binding substance adsorbed on a large number of adsorptive regions 4 with a chemiluminescent substrate, and a fluorescent substance such as a fluorescent dye A biologically-derived substance labeled with is selectively hybridized.
[0091]
In this way, chemiluminescence data and fluorescence data are recorded in a large number of the adsorptive regions 4 of the biochemical analysis unit 1.
[0092]
FIG. 4 is a schematic cross-sectional view of a biochemical analysis data generating apparatus according to a preferred embodiment of the present invention.
[0093]
The biochemical analysis data generation apparatus according to the present embodiment is configured to be able to read fluorescence data recorded in a large number of adsorptive regions 4 of the biochemical analysis unit 1, as shown in FIG. A cooling CCD area sensor 20, an excitation light cut filter member 21 disposed in front of the cooling CCD area sensor 20, a sample stage 25 having a transparent glass plate 24 on which the biochemical analysis unit 1 is placed, and a sample A lens array 23 in which a plurality of convex lenses 22 are formed at positions corresponding to a large number of dot-like adsorptive regions 4 of the biochemical analysis unit 1 placed close to the stage 25 and placed on the sample stage 25. And a laser excitation light source 27 that emits laser light 26 and a laser light 26 emitted from the laser excitation light source 20 are diverged to obtain a sample stage 25. And a concave lens 28 to be incident on the biochemical analysis unit 1 placed on the glass plate 24.
[0094]
As the laser excitation light source 27, a laser excitation light source that emits a laser beam 26 having a wavelength capable of efficiently exciting a fluorescent substance contained in a large number of adsorptive regions 4 formed in the biochemical analysis unit 1 is selected. For example, when Cy3 (registered trademark) is used as the fluorescent material, a laser excitation light source 27 that emits laser light 26 having a wavelength of 473 nm is used.
[0095]
Therefore, in this embodiment, the excitation light cut filter member 21 has a property of cutting light having a wavelength of 473 nm, which is equal to the wavelength of the laser light 26 emitted from the laser excitation light source 27, and transmitting light having a wavelength longer than 473 nm. Have.
[0096]
FIG. 5 is a block diagram showing a control system, a detection system and a memory system of the cooled CCD area sensor 20, and a control system, a memory system, a display system and an input system of the biochemical analysis data generating apparatus according to this embodiment. .
[0097]
As shown in FIG. 5, the cooled CCD area sensor 20 includes a CCD 30, an A / D converter 31 that digitizes analog data accumulated in the form of charges by the CCD 30, and an A / D converter 31. A data buffer 32 that temporarily stores the digitized and generated biochemical analysis data and a camera control circuit 33 that controls the operation of the cooled CCD area sensor 20 are provided.
[0098]
As shown in FIG. 5, the biochemical analysis data generating apparatus according to this embodiment includes a CPU 40 that controls the operation of the cooled CCD area sensor 20 and biochemical analysis data generated by the cooled CCD area sensor 20. The data transfer means 41 for reading the data from the data buffer 32, the data processing means 42 for performing data processing on the biochemical analysis data read by the data transfer means 41, and the data processing means 42 for performing data processing. Data storage means 43 for storing the biochemical analysis data, and data display means 44 for generating quantitative data based on the biochemical analysis data stored in the data storage means 43 and displaying it on the screen of the CRT 48 And a light source control means 45 for controlling the laser excitation light source 27 and a keyboard 46 for inputting various instruction signals. There. The CPU 40 is configured to control the light source control means 45 based on the instruction signal input via the keyboard 46 and to output various signals to the camera control circuit 33 of the cooling CCD area sensor 20.
[0099]
The biochemical analysis data generating apparatus according to the present embodiment configured as described above has the fluorescence recorded in the dot-like adsorptive region 4 formed in the biochemical analysis unit as follows. Read data and generate data for biochemical analysis.
[0100]
First, a biochemical analysis unit 1 in which fluorescence data is recorded in a large number of dot-like adsorptive regions 4 is placed on a transparent glass plate 24 of a sample stage 25 by a user.
[0101]
Here, the sample stage 25 is provided with a guide member (not shown), and a large number of dot-like adsorptive regions 4 are respectively opposed to the corresponding convex lenses 22 formed in the lens array 23. It is guaranteed that the biochemical analysis unit 1 is placed on the sample stage 25.
[0102]
Next, when a data generation start signal is input to the keyboard 46 by the user, the data generation start signal is input to the CPU 40.
[0103]
When receiving the data generation start signal, the CPU 40 outputs a start signal to the laser excitation light source 27 to start the laser excitation light source 27.
[0104]
The laser beam 26 having a wavelength of 473 nm emitted from the laser excitation light source 27 is incident on the concave lens 28, is diverged, and is placed on the entire surface of the biochemical analysis unit 1 placed on the glass plate 24 of the sample stage 25. Laser light 26 is irradiated.
[0105]
When irradiated with the laser beam 26, the fluorescent material, for example, Cy3 contained in each adsorptive region 4 is excited and the fluorescence 29 is emitted.
[0106]
The fluorescence 29 emitted from the dot-like adsorptive regions 4 is incident on the convex lens 22 that faces each of the adsorptive regions 4 and is disposed in close proximity, and collects in the opposing region of the excitation light cut filter member 21. Lighted.
[0107]
Here, the fluorescent material is excited by the excitation light, and the wavelength of the emitted fluorescence 29 is longer than the wavelength of the excitation light. In this embodiment, the excitation light cut filter member 21 emits from the laser excitation light source 27. Since the light having a wavelength of 473 nm, which is equal to the wavelength of the laser beam 26, is cut and the light having a wavelength longer than 473 nm is transmitted, the light having a wavelength of 473 nm is cut and included in the adsorptive region 4 Only the fluorescence 29 emitted from the fluorescent material is transmitted through the excitation light cut filter member 21 and enters the photocathode of the CCD 30 to form an image on the photocathode.
[0108]
The CCD 30 thus receives the light of the image formed on the photocathode and accumulates it in the form of charges.
[0109]
When a predetermined exposure time has elapsed, the CPU 40 outputs an exposure completion signal to the camera control circuit 33 and the light source control means 45 of the cooling CCD area sensor 20.
[0110]
The light source controller 45 turns off the laser excitation light source 27 when receiving an exposure completion signal from the CPU 40.
[0111]
On the other hand, when receiving an exposure completion signal from the CPU 40, the camera control circuit 33 transfers analog data accumulated in the form of charges by the CCD 30 to the A / D converter 31, digitizes it, and generates biochemical analysis data. Then, it is temporarily stored in the data buffer 32.
[0112]
At the same time, the CPU 40 outputs a data transfer signal to the data transfer means 41, reads the biochemical analysis data from the data buffer 32 of the cooled CCD area sensor 20, and causes the data processing means 42 to output it.
[0113]
The data processing means 42 performs necessary data processing on the input biochemical analysis data in accordance with a user instruction, and stores the data in the data storage means 43.
[0114]
When the user inputs a data display signal to the keyboard 46, the CPU 40 outputs a data display signal to the data display means 44, and generates quantitative data based on the biochemical analysis data stored in the data storage means 43. , Displayed on the screen of the CRT 48.
[0115]
In the present embodiment, the laser light 26 emitted from the laser excitation light source 27 is diverged by the concave lens 28 and is applied to the entire surface of the biochemical analysis unit 1 placed on the glass plate 24 of the sample stage 25. The light 26 is irradiated, the fluorescent substance contained in the adsorptive region 4 formed in a dot shape in the biochemical analysis unit 1 is excited, and the fluorescence 29 is emitted. The lens array 23 is arranged in the vicinity of the biochemical analysis unit 1 placed on the sample stage 25, and is located at a position facing each adsorptive region 4 formed in a dot shape on the biochemical analysis unit 1. Since each has the convex lens 22, the fluorescence 29 emitted from each adsorptive region 4 is condensed on the corresponding region of the excitation light cut filter member 21 by the corresponding convex lens 22 of the lens array 23, and the laser beam. Only the fluorescent light 29 having a wavelength longer than that of the laser light 26 is transmitted through the excitation light cut filter member 21 and is detected photoelectrically by the CCD 30 of the cooled CCD area sensor 20. The data is digitized by the / D converter 31 to generate biochemical analysis data.
[0116]
Therefore, according to this embodiment, even when the dot-like adsorptive regions 4 are formed in the biochemical analysis unit 1 with high density, only the fluorescence 29 emitted from each of the adsorptive regions 4 is opposed. Since the light is condensed by the convex lens 22 and guided to the CCD 30 of the cooled CCD area sensor 20 through the excitation light cut filter 21, the fluorescence 29 is detected with high sensitivity and data for biochemical analysis is generated. It becomes possible.
[0117]
FIG. 6 is a schematic cross-sectional view of a biochemical analysis data generating apparatus according to another preferred embodiment of the present invention.
[0118]
The biochemical analysis data generation apparatus according to the present embodiment is configured to be able to read chemiluminescence data recorded in a large number of adsorptive regions 4 of the biochemical analysis unit 1, as shown in FIG. In addition, a cooled CCD area sensor 50, a sample stage 55 provided with a transparent glass plate 54 on which the biochemical analysis unit 1 is placed, and a raw stage placed on the sample stage 55 and placed in the vicinity of the sample stage 55. A lens array 53 in which a plurality of convex lenses 52 are formed is provided at positions corresponding to a large number of dot-like adsorptive regions 4 of the chemical analysis unit 1.
[0119]
FIG. 7 is a block diagram showing a control system, a detection system and a memory system of the cooled CCD area sensor 50, and a control system, a memory system, a display system and an input system of the biochemical analysis data generating apparatus according to this embodiment. .
[0120]
As shown in FIG. 7, the cooled CCD area sensor 50 includes a CCD 60, an A / D converter 61 that digitizes analog data accumulated in the form of electric charges by the CCD 60, and an A / D converter 61. A data buffer 62 that temporarily stores the digitized and generated biochemical analysis data and a camera control circuit 63 that controls the operation of the cooling CCD area sensor 50 are provided.
[0121]
As shown in FIG. 7, the biochemical analysis data generating apparatus according to this embodiment includes a CPU 70 that controls the operation of the cooled CCD area sensor 50, and biochemical analysis data generated by the cooled CCD area sensor 50. The data transfer means 71 for reading out the data from the data buffer 62, the data processing means 72 for performing data processing on the biochemical analysis data read by the data transfer means 71, and the data processing by the data processing means 72. Data storage means 73 for storing the biochemical analysis data, and data display means 74 for generating quantitative data based on the biochemical analysis data stored in the data storage means 43 and displaying it on the screen of the CRT 78 A keyboard 76 for inputting various instruction signals is provided. The CPU 70 is configured to be able to output various signals to the camera control circuit 63 of the cooling CCD area sensor 50 based on the instruction signal input via the keyboard 76.
[0122]
The biochemical analysis data generating apparatus according to the present embodiment configured as described above has the chemistry recorded in the dot-like adsorptive region 4 formed in the biochemical analysis unit as follows. Reads luminescence data and generates data for biochemical analysis.
[0123]
First, a biochemistry in which a chemiluminescent substrate is brought into contact with a labeling substance contained in a large number of dot-like porous regions 4 formed in the biochemical analysis unit 1 by a user to emit chemiluminescence 59. The analysis unit 1 is placed on the transparent glass plate 54 of the sample stage 55.
[0124]
Here, the sample stage 55 is provided with a guide member (not shown), and a large number of dot-like adsorptive regions 4 are respectively opposed to the corresponding convex lenses 52 formed in the lens array 53. It is guaranteed that the biochemical analysis unit 1 is placed on the sample stage 55.
[0125]
Next, when a data generation start signal is input to the keyboard 76 by the user, the data generation start signal is input to the CPU 70.
[0126]
When the data generation start signal is input, the CPU 70 outputs the data generation start signal to the camera control circuit 63 of the cooling CCD area sensor 50, and is formed in the biochemical analysis unit 1 in the cooling CCD area sensor 50. Detection of chemiluminescence 59 emitted from the dot-like adsorptive region 4 is started.
[0127]
The chemiluminescence 59 emitted from the dot-shaped adsorptive regions 4 respectively enter the convex lenses 52 arranged in close proximity to the adsorbing regions 4, and the photocathode of the CCD 60 of the cooled CCD area sensor 50. To form an image on the photocathode.
[0128]
The CCD 60 of the cooled CCD area sensor 50 thus receives the light of the image formed on the photocathode and accumulates it in the form of charges.
[0129]
When a predetermined exposure time has elapsed, the CPU 70 outputs an exposure completion signal to the camera control circuit 63 of the cooling CCD area sensor 50.
[0130]
Upon receiving an exposure completion signal from the CPU 40, the camera control circuit 33 transfers the analog data accumulated in the form of charges by the CCD 30 to the A / D converter 31, digitizes it, and generates biochemical analysis data. The data buffer 32 is temporarily stored.
[0131]
At the same time, the CPU 40 outputs a data transfer signal to the data transfer means 41, reads the biochemical analysis data from the data buffer 32 of the cooled CCD area sensor 20, and causes the data processing means 42 to output it.
[0132]
The data processing means 42 performs necessary data processing on the input biochemical analysis data in accordance with a user instruction, and stores the data in the data storage means 43.
[0133]
When the user inputs a data display signal to the keyboard 46, the CPU 40 outputs a data display signal to the data display means 44, and generates quantitative data based on the biochemical analysis data stored in the data storage means 43. , Displayed on the screen of the CRT 78.
[0134]
According to the present embodiment, even when the dot-like adsorptive regions 4 are formed in the biochemical analysis unit 1 at a high density, each adsorptive region formed in the biochemical analysis unit 1 in a dot shape. 4, the lens array 53 having the convex lens 22 is provided in the vicinity of the biochemical analysis unit 1 placed on the sample stage 55 at a position opposite to the lens 4. Since only the chemiluminescence 59 that has been collected can be collected by the opposing convex lens 52 and guided to the CCD 60 of the cooled CCD area sensor 50, the chemiluminescence 59 is detected with high sensitivity and data for biochemical analysis is generated. It becomes possible to do.
[0135]
FIG. 8 is a schematic perspective view of a biochemical analysis unit used in a method for generating biochemical analysis data according to another preferred embodiment of the present invention.
[0136]
As shown in FIG. 8, the biochemical analysis unit 80 according to this embodiment includes an aluminum substrate 81 in which a large number of substantially circular through holes 82 are regularly formed. An adsorbent film 83 formed of nylon 6 is press-fitted into the through-hole 82 by a calendar processing device (not shown), and a large number of adsorbent regions 84 are regularly formed in a dot shape. ing.
[0137]
Although not shown exactly in FIG. 8, in this embodiment, approximately circular absorptive regions 84 having a size of about 0.01 square millimeters of about 10000 at a density of about 5000 pieces / square centimeter, The biochemical analysis unit 80 is regularly formed.
In this embodiment, the adsorptive film 83 is press-fitted into the through hole 82 formed in the substrate 81 so that the surface of the adsorptive region 84 and the surface of the substrate 81 are located at the same height. A biochemical analysis unit 80 is formed.
[0138]
Also in this embodiment, in the same manner as in the biochemical analysis unit 1 according to the above-described embodiment shown in FIG. 1, the spotting device 5 applies a large number of adsorptive regions 84 formed in the biochemical analysis unit 80. Specific binding substances such as cDNA are dropped and adsorbed.
[0139]
Furthermore, as shown in FIG. 3, a hybridizing solution 9 containing a biological substance labeled with a labeling substance that produces chemiluminescence by contacting with a chemiluminescent substrate and a biological substance labeled with a fluorescent substance 9 The biochemical analysis unit 80 is set in the hybridization container 8 containing the chemical substance, and chemiluminescence is produced by contacting a specific binding substance such as cDNA adsorbed on a number of the adsorptive regions 84 with a chemiluminescent substrate. Labeled with a labeling substance to be generated, labeled with a biological substance and a fluorescent substance contained in the hybridizing solution 9, and selectively hybridized with a biological substance contained in the hybridizing liquid 9.
[0140]
In this way, chemiluminescence data and fluorescence data are recorded in the biochemical analysis unit 80.
[0141]
The fluorescence data recorded in the biochemical analysis unit 80 is read by the biochemical analysis data generator shown in FIGS. 4 and 5 in the same manner as in the above-described embodiment. The chemiluminescence data recorded in the unit 80 is read by the biochemical analysis data generator shown in FIGS. 6 and 7 in the same manner as in the above embodiment, and the biochemical analysis data is generated respectively. Is done.
[0142]
The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope of the invention described in the claims, and these are also included in the scope of the present invention. Needless to say.
[0143]
For example, in the above-described embodiment, a plurality of different cDNAs having known base sequences are used as the specific binding substance, but the specific binding substance usable in the present invention is not limited to cDNA. It can specifically bind to biological substances such as hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA, etc., and the base sequence and length of the base Any specific binding substance whose composition is known can be used as the specific binding substance of the present invention.
[0144]
Further, in the embodiment shown in FIGS. 4 and 5, a laser excitation light source 27 that emits a laser beam 26 having a wavelength of 473 nm is used, so that the light having a wavelength of 473 nm is cut and the wavelength is longer than 473 nm. Although the excitation light cut filter 21 having the property of transmitting light is used, the laser excitation that emits the laser light 26 that can efficiently excite the fluorescent substance according to the fluorescent substance that labels the substance derived from the living body. The light source 27 can be used, and instead of the laser excitation light source 27 that emits the laser beam 26 having a wavelength of 473 nm, for example, a second harmonic generation element that emits a laser beam having a wavelength of 532 nm, A semiconductor laser excitation light source that emits laser light having a wavelength can also be used.
[0145]
In the embodiment shown in FIGS. 4 and 5, the laser excitation light source 27 that emits the laser light 26 having a wavelength of 473 nm is used. However, it is not always necessary to use a laser excitation light source as the excitation light source. In place of the laser excitation light source, an LED light source may be used as the excitation light source. Further, a halogen lamp may be used as the excitation light source, and a wavelength component that does not contribute to excitation may be cut by a spectral filter. .
[0146]
Further, in the embodiment shown in FIGS. 4 and 5, the concave lens 28 is used to diverge the laser light 2 emitted from the laser excitation light source 27, and the entire surface of the biochemical analysis unit 1 is irradiated with the laser light 26. However, the means for diverging the laser beam 26 to irradiate the entire surface of the biochemical analysis unit 1 can be arbitrarily selected, and the concave lens 28 is necessarily used. Not.
[0147]
In the above embodiment, the cooled CCD area sensors 20 and 50 are used as the photodetectors. However, instead of the cooled CCD area sensor, a CCD area sensor that does not have a cooling function may be used. Further, instead of the CCD, other solid-state sensors such as a CID (charge injection element), a PDA (photodiode array), and a MOS type image pickup element can be used.
[0148]
Furthermore, in the embodiment shown in FIG. 1, the biochemical analysis unit 1 is formed by filling nylon 6 into a large number of through holes 3 formed in an aluminum substrate 2. In the embodiment shown in FIG. 8, the biochemical analysis unit 80 includes a large number of adsorbent films 83 formed of nylon 6 and formed on an aluminum substrate 81. The adsorbing region 84 of the biochemical analysis unit 1 and the adsorbing region 84 of the biochemical analysis unit 80 are made of nylon. 6 is not necessarily required. Instead of nylon 6, other porous material capable of forming a membrane filter, for example, nylon 6, 6, nylon 4, 10, etc. Ron; Cellulose derivatives such as nitrocellulose, cellulose acetate, and cellulose butyrate; Collagen; Alginates such as alginic acid, calcium alginate, alginic acid / polylysine polyion complex; Polyolefins such as polyethylene and polypropylene; Polyvinyl chloride; Polyvinylidene chloride; The adsorptive region 4 of the biochemical analysis unit 1 and the adsorptive region 84 of the biochemical analysis unit 80 are formed by polyfluoride such as polyvinylidene fluoride and polytetrafluoride, or a copolymer or complex thereof. Furthermore, carbon porous materials such as activated carbon or metals such as platinum, gold, iron, silver, nickel and aluminum; metal oxides such as alumina, silica, titania and zeolite; hydroxyapata The absorptive region 4 of the biochemical analysis unit 1 and the absorptive region 84 of the biochemical analysis unit 80 are made of an inorganic porous material such as a metal salt such as calcium sulfate or a composite thereof or a bundle of a plurality of fibers. May be formed.
[0149]
Furthermore, in the said embodiment, although the units 1 and 80 for biochemical analysis are equipped with the board | substrates 2 and 81 made from aluminum, the board | substrates 2 and 81 of the units 1 and 80 for biochemical analysis are made into aluminum. However, it is preferable that the substrates 2 and 81 of the biochemical analysis units 1 and 80 are made of a material that attenuates light. However, the material is particularly limited. Any of inorganic compound materials and organic compound materials can be used, and metal materials, ceramic materials or plastic materials are particularly preferably used. Examples of inorganic compound materials that can be preferably used for forming the substrates 2 and 81 of the biochemical analysis units 1 and 80 include gold, silver, copper, zinc, aluminum, titanium, tantalum, chromium, iron, nickel, Metals such as cobalt, lead, tin, and selenium; alloys such as brass, stainless steel, and bronze; silicon materials such as silicon, amorphous silicon, glass, quartz, silicon carbide, and silicon nitride; metals such as aluminum oxide, magnesium oxide, and zirconium oxide Examples of oxides include inorganic salts such as tungsten carbide, calcium carbonate, calcium sulfate, hydroxyapatite, and gallium arsenide. These may have any structure in a polycrystalline sintered body such as single crystal, amorphous, or ceramic. In addition, as an organic compound material that can be preferably used for forming the substrates 2 and 81 of the biochemical analysis units 1 and 80, a polymer compound is preferably used, and examples of preferable polymer compounds include polyethylene and polypropylene. Acrylic resins such as polymethyl methacrylate and butyl acrylate / methyl methacrylate copolymer; polyacrylonitrile; polyvinyl chloride; polyvinylidene chloride; polyvinylidene fluoride; polytetrafluoroethylene; polychlorotrifluoroethylene; Polyesters such as naphthalate and polyethylene terephthalate; nylons such as nylon 6, nylon 6,6, nylon 4,10; polyimide; polysulfone; polyphenylene sulfide; Silicon resin such as lydiphenylsiloxane; phenolic resin such as novolac; epoxy resin; polyurethane; polystyrene; butadiene-styrene copolymer; polysaccharide such as cellulose, cellulose acetate, nitrocellulose, starch, calcium alginate, hydroxypropyl methylcellulose; Chitosan; urushi; polyamides such as gelatin, collagen, keratin, and copolymers of these polymer compounds. These may be composite materials, and if necessary, may be filled with metal oxide particles or glass fibers, or may be used by blending organic compound materials.
[0150]
Further, in the embodiment shown in FIG. 8, the absorptive region 84 of the biochemical analysis unit 80 has a large number of penetrations formed in the aluminum substrate 81 through the absorptive film 83 formed of nylon 6. Although it is formed by press-fitting into the hole 82, it is not always necessary to press-fit the adsorptive film 83 into the large number of through-holes 82 formed in the substrate 81 using a calendar processing apparatus. The adsorptive film 83 can be press-fitted into a large number of through-holes 82 formed in the substrate 81 by using other means such as a press device. Instead of press-fitting, the adsorbent film 83 is obtained by an appropriate method. May be embedded in a large number of through-holes 82 formed in the substrate 81 to form a large number of adsorptive regions 4.
[0151]
Further, in the above-described embodiment, the substrate 2, 81 of the biochemical analysis unit 1, 80 has approximately circular absorptive regions 4, 84 having a size of about 0.01 square millimeters of about 10,000, about 5000 It is formed on the substrates 2 and 81 according to a regular pattern with a density of pieces / square centimeter, but it is not always necessary to form the adsorptive regions 4 and 84 in a substantially circular shape, such as a rectangular shape, It can be formed in any shape.
[0152]
In the above embodiment, the substrates 2 and 81 of the biochemical analysis units 1 and 80 have approximately circular absorptive regions 4 and 84 having a size of about 0.01 square millimeters of about 10,000 and about 5000. The number of the adsorbing regions 4 and 84 and the size thereof are arbitrarily selected according to the purpose. Preferably, the adsorptive regions 4 and 84 having a size of 10 or more and less than 5 square millimeters are formed on the substrate 2 and 81 at a density of 10 or more per square centimeter.
[0153]
Further, in the above-described embodiment, the substrate 2, 81 of the biochemical analysis unit 1, 80 has approximately circular absorptive regions 4, 84 having a size of about 0.01 square millimeters of about 10,000, about 5000 Although formed in a regular pattern at a density of pcs / square centimeter, it is not necessary to form the adsorptive regions 4, 84 according to a regular pattern.
[0154]
In the above embodiment, a hybridizing solution 9 containing a labeling substance that generates chemiluminescence by contacting with a chemiluminescent substrate and a biological substance labeled with a fluorescent substance is prepared and dropped into the adsorptive region 4. A biologically-derived substance labeled with a fluorescent substance and a biologically-derived substance labeled with a labeling substance that is hybridized to the specific binding substance that has been produced, but generates chemiluminescence by contacting with the chemiluminescent substrate, It is not always necessary to selectively hybridize with specific binding substances contained in a large number of the adsorptive regions 4 of the biochemical analysis unit 1 at the same time. A biological substance labeled with a fluorescent substance and a biological substance labeled with a fluorescent substance One quality, the specific binding substances contained in a number of the absorptive regions 4 of the biochemical analysis unit 1, selectively, it may be allowed to hybridize.
[0155]
Furthermore, in the above-described embodiment, a biological substance labeled with a labeling substance that generates chemiluminescence upon contact with a chemiluminescent substrate and a biological substance labeled with a fluorescent substance hybridize to the specific binding substance. Although it is soyed, it is not always necessary to hybridize a substance derived from a living body to a specific binding substance. Instead of hybridization, a substance derived from a living body is subjected to a reaction such as an antigen-antibody reaction or a receptor / ligand. Alternatively, it can be specifically bound to a specific binding substance.
[0156]
Furthermore, in the said embodiment, the spotting apparatus provided with the injector 6 and the CCD camera 7 is used, and the adsorbing area | region 4 which should drop a specific binding substance, such as cDNA, with the CCD camera 7 by the CCD camera 7 is used. While observing, when the tip of the injector 6 and the center of the adsorptive region 4 to which the specific binding substance such as cDNA is dropped coincide with each other, the specific binding substance such as cDNA is released from the injector 6. The relative positional relationship between the tip of the injector 6 and the numerous adsorptive regions 4 formed in the biochemical analysis unit 1 is detected in advance, and the injector 6 The chemical analysis unit 1 is moved two-dimensionally at a relatively constant pitch so that a specific binding substance such as cDNA is dropped. It can also be.
[0157]
【The invention's effect】
According to the present invention, a specific binding substance that can be specifically bound to a substance derived from a living body and has a known base sequence, base length, composition, and the like is dropped onto a carrier in a spot shape to increase the amount. In the density, a spot-like region is formed, and a specific binding substance contained in the spot-like region is brought into contact with a chemiluminescent substrate to cause chemiluminescence, and is derived from a biological substance labeled with a labeling substance and / or a fluorescent substance. Biochemical analysis data is generated by photoelectrically detecting chemiluminescence and / or fluorescence emitted from a biochemical analysis unit obtained by specifically binding and selectively labeling substances Even when analyzing substances of this type, it is possible to effectively prevent noise caused by chemiluminescence and / or fluorescence scattering from being generated in the data for biochemical analysis. Generate data It becomes possible to provide a production method and apparatus biochemical analysis data that can Rukoto.
[Brief description of the drawings]
FIG. 1 is a schematic perspective view of a biochemical analysis unit used in a method for generating biochemical analysis data according to a preferred embodiment of the present invention.
FIG. 2 is a schematic front view of a spotting device.
FIG. 3 is a schematic longitudinal sectional view of a hybridization container.
FIG. 4 is a schematic cross-sectional view of a biochemical analysis data generating apparatus according to a preferred embodiment of the present invention.
FIG. 5 is a block diagram showing a control system, a detection system, and a memory system of a cooled CCD area sensor, and a control system, a memory system, a display system, and an input system of a biochemical analysis data generation apparatus according to the present embodiment; It is a diagram.
FIG. 6 is a schematic cross-sectional view of a biochemical analysis data generating apparatus according to another preferred embodiment of the present invention.
FIG. 7 is a block diagram illustrating a control system, a detection system, and a memory system of a cooled CCD area sensor, and a control system, a memory system, a display system, and an input system of a biochemical analysis data generation apparatus according to the present embodiment; It is a diagram.
FIG. 8 is a schematic perspective view of a biochemical analysis unit used in a method for generating biochemical analysis data according to another preferred embodiment of the present invention.
[Explanation of symbols]
1 Biochemical analysis unit
2 Substrate
3 Through hole
4 Adsorbent area
5 Spotting device
6 Injector
7 CCD camera
8 Hybridization container
9 Hybridization solution
20 Cooling CCD area sensor
21 Excitation light cut filter member
22 Convex lens
23 Lens array
24 Transparent glass plate
25 Sample stage
26 Laser light
27 Laser excitation light source
28 Concave lens
29 Fluorescence
30 CCD
31 A / D converter
32 data buffers
33 Camera control circuit
40 CPU
41 Data transfer means
42 Data processing means
43 Data storage means
44 Data display means
45 Light source control means
46 keyboard
48 CRT
50 Cooling CCD area sensor
52 Convex lens
53 Lens Array
54 Transparent glass plate
55 Sample stage
59 Chemiluminescence
60 CCD
61 A / D converter
62 Data buffer
63 Camera control circuit
70 CPU
71 Data transfer means
72 Data processing means
73 Data storage means
74 Data display means
76 keyboard
78 CRT
80 Biochemical analysis unit
81 substrates
82 Through hole
83 Adsorbent membrane
84 Adsorbent area

Claims (25)

A plurality of holes are two-dimensionally formed by being spaced apart from each other and two-dimensionally formed by filling the plurality of holes in the substrate with an adsorbent material and being separated from each other. A unit for biochemical analysis having a plurality of adsorptive regions is placed on a sample stage, and light emitted from the plurality of adsorptive regions of the unit for biochemical analysis placed on the sample stage, The two-dimensional solid sensor condenses light by a plurality of condensing lenses arranged opposite to each of the plurality of absorptive regions of the biochemical analysis unit, and is detected photoelectrically by the two-dimensional solid sensor. Then, a method for generating biochemical analysis data, characterized by generating biochemical analysis data.   The biochemical analysis data generation method according to claim 1, wherein a substrate of the biochemical analysis unit has a property of attenuating light.   The substrate of the biochemical analysis unit has a property of attenuating light energy to 1/5 or less when light is transmitted through the substrate by a distance equal to the distance between adjacent adsorptive regions. The method for generating data for biochemical analysis according to claim 2.   6. The biochemical analysis according to claim 2, wherein the substrate of the biochemical analysis unit is formed of a material selected from the group consisting of a metal material, a ceramic material, and a plastic material. Data generation method.   The method for generating data for biochemical analysis according to any one of claims 1 to 4, wherein the condensing lens is constituted by a lens having a large numerical aperture.   The plurality of condensing lenses are attached to a lens array so that a pitch between adjacent condensing lenses is equal to a pitch between adjacent adsorptive regions of the biochemical analysis unit. The method for generating data for biochemical analysis according to any one of 1 to 5.   The method for generating data for biochemical analysis according to any one of claims 1 to 6, wherein the two-dimensional solid-state sensor is constituted by a cooled CCD sensor.   The plurality of absorptive regions of the biochemical analysis unit each include a specific binding substance, and the specific binding substance contained in the plurality of absorptive regions is brought into contact with a chemiluminescent substrate to emit chemiluminescence. A biologically-derived substance labeled with a labeling substance that generates luminescence is selectively and specifically bound, and chemiluminescence emitted from the plurality of adsorptive regions is photoelectrically detected by the two-dimensional solid sensor. The data for biochemical analysis according to any one of claims 1 to 7, wherein the data for biochemical analysis is generated.   The plurality of absorptive regions of the biochemical analysis unit each contain a specific binding substance, and the biologically-derived substance labeled with a fluorescent substance on the specific binding substance contained in the plurality of absorptive regions Selectively emitted from an excitation light source located on the opposite side of the two-dimensional solid sensor with respect to the sample stage in the plurality of absorptive regions of the biochemical analysis unit. The excitation light is irradiated to excite the fluorescent material selectively contained in the plurality of adsorptive regions, and the fluorescence emitted from the fluorescent material is converted into the wavelength of the excitation light by the plurality of condenser lenses. The light is cut and guided to the two-dimensional sensor through an excitation hole cut filter having a property of transmitting light having a wavelength longer than that of the excitation light. Chemical solution The method of generating the biochemical analysis data according to any one of claims 1 to 7, characterized in that to produce a use data.   10. The biologically-derived substance is bound to the specific binding substance by a reaction selected from the group consisting of hybridization, an antigen-antibody reaction, and a receptor / ligand. How to generate data for biochemical analysis.   The biochemical analysis data generation method according to any one of claims 1 to 10, wherein 10 or more adsorptive regions are formed on the substrate of the biochemical analysis unit.   The biochemical analysis unit according to any one of claims 1 to 11, wherein each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 5 square millimeters. How to generate data.   13. The plurality of absorptive regions are formed on the substrate of the biochemical analysis unit at a density of 10 pieces / square centimeter or more, according to any one of claims 1 to 12. How to generate data for biochemical analysis. A plurality of through holes are two-dimensionally formed on the substrate of the biochemical analysis unit so as to be separated from each other, and the plurality of adsorptive regions are filled with an adsorbent material in the plurality of through holes. The method for generating data for biochemical analysis according to any one of claims 1 to 13, wherein the data is generated. A plurality of through holes are two-dimensionally formed in the substrate of the biochemical analysis unit so as to be separated from each other, and the plurality of adsorptive regions of the biochemical analysis unit include an adsorbent material. The biochemical analysis data generation method according to claim 14, wherein the conductive film is formed by being press-fitted into the plurality of through holes of the substrate.   16. The generation of data for biochemical analysis according to any one of claims 1 to 15, wherein the plurality of adsorptive regions are regularly formed on the substrate of the biochemical analysis unit. Method.   17. The raw material according to claim 1, wherein the adsorptive region of the biochemical analysis unit is formed of a porous carbon material or a porous material capable of forming a membrane filter. How to generate data for chemical analysis.   The biochemical analysis data generation method according to any one of claims 1 to 16, wherein the adsorptive region of the biochemical analysis unit is formed by a bundle of a plurality of fibers.   The method for generating data for biochemical analysis according to any one of claims 1 to 18, wherein the excitation light source is a laser excitation light source. A plurality of holes are two-dimensionally formed by being spaced apart from each other and two-dimensionally formed by filling the plurality of holes in the substrate with an adsorbent material and being separated from each other. A sample stage on which a biochemical analysis unit having a plurality of adsorptive regions is placed; a two-dimensional solid sensor; and the plurality of absorptive properties of the biochemical analysis unit placed on the sample stage An apparatus for generating data for biochemical analysis, comprising a lens array having a plurality of condensing lenses attached at positions facing each of the regions.   Further, an excitation light source that emits excitation light is provided on the opposite side of the two-dimensional solid state sensor with respect to the sample stage, and light having the wavelength of the excitation light is cut between the lens array and the two-dimensional solid state sensor. 21. The apparatus for generating biochemical analysis data according to claim 20, further comprising an excitation light cut filter having a property of transmitting light having a wavelength longer than that of the excitation light.   The apparatus for generating data for biochemical analysis according to claim 20 or 21, wherein the condensing lens is constituted by a lens having a large numerical aperture.   The plurality of condensing lenses are attached to the lens array so that a pitch between adjacent condensing lenses is equal to a pitch between adjacent adsorbing regions of the biochemical analysis unit. Item 23. The apparatus for generating data for biochemical analysis according to any one of Items 20 to 22.   24. The biochemical analysis data generation apparatus according to claim 20, wherein the two-dimensional solid sensor is a cooled CCD sensor.   25. The apparatus for generating biochemical analysis data according to claim 20, wherein the excitation light source is a laser excitation light source.

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