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WO2004081570A1 - Matrice de puce, puce comprenant une matrice et leur preparation et application - Google Patents

Matrice de puce, puce comprenant une matrice et leur preparation et application Download PDF

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Publication number
WO2004081570A1
WO2004081570A1 PCT/CN2003/001091 CN0301091W WO2004081570A1 WO 2004081570 A1 WO2004081570 A1 WO 2004081570A1 CN 0301091 W CN0301091 W CN 0301091W WO 2004081570 A1 WO2004081570 A1 WO 2004081570A1
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WO
WIPO (PCT)
Prior art keywords
chip
substrate
pigments
black
light
Prior art date
Application number
PCT/CN2003/001091
Other languages
English (en)
Chinese (zh)
Inventor
Fanglin Zou
Chunsheng Chen
Ning Chen
Jianxia Wang
Original Assignee
Chengdu Kuachang Medical Industrial Limited
Chengdu Kuachang Science & Technology Co., Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from CN 03117446 external-priority patent/CN1250969C/zh
Priority claimed from CNB031176453A external-priority patent/CN100347545C/zh
Priority claimed from CNA031177875A external-priority patent/CN1514243A/zh
Application filed by Chengdu Kuachang Medical Industrial Limited, Chengdu Kuachang Science & Technology Co., Ltd filed Critical Chengdu Kuachang Medical Industrial Limited
Priority to AU2003289617A priority Critical patent/AU2003289617A1/en
Priority to CN200480000649.8A priority patent/CN1735807A/zh
Priority to PCT/CN2004/000713 priority patent/WO2005059553A1/fr
Priority to PCT/CN2004/000839 priority patent/WO2005083432A1/fr
Publication of WO2004081570A1 publication Critical patent/WO2004081570A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54346—Nanoparticles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00158—Elements containing microarrays, i.e. "biochip"

Definitions

  • the invention relates to a detection chip substrate for qualitative and / or quantitative analysis of a target object in a sample, especially a biological sample, and also relates to a detection chip including the substrate, a detection chip scanning device, and a detection chip.
  • a detection chip including the substrate, a detection chip scanning device, and a detection chip. The preparation method of the substrate and chip is described. Background technique
  • the “detection chip” in the present invention also referred to as “chip” for short, includes, but is not limited to, “Biochip”, “Microarray”, and “Bioarray” in English. It is a detection device in a specific and / or quantitative analysis. The specific reaction between the trace probe in the reactor and the target molecule in the sample can be identified in an addressable manner.
  • the core of the chip is the reactor in which the core of the reactor is the chip base and the chip fixed on the chip
  • the distribution density of the probes on the substrate is greater than 10 dots / cm 2 , preferably greater than 20 dots / cm 2 , and more preferably greater than 40 dots / cm 2 , and each The area of the probe point is not greater than 1 mm
  • Chips include biochips and non-biochips. Currently, the most commonly used are biochips. The most commonly used biochips are peptide chips and gene chips.
  • the peptide chip is a biochip prepared by immobilizing a sequence structure of multiple amino acids (including proteins) on a substrate as a probe.
  • a gene chip is a chip that hybridizes nucleic acids, nucleotides with complementary nucleic acids, and nucleotide probes in a sample to be tested to form a hybrid, or binds to a specific antibody, and then displays the detection result with a color reaction.
  • Biochips have a wide range of applications, including gene expression detection, gene screening, drug screening, disease diagnosis and treatment, environmental monitoring and governance, and judicial identification.
  • Biochip probes include all biologically active substances that can be immobilized on a solid support, such as antigens, antibodies, single- and multi-stranded DNA, RNA, nucleotides, ligands, ligands, peptides, cells, Tissue components and other biological components.
  • the chip substrate in the present invention refers to a product based on the chip base and optionally combined with other structures (such as an isolation structure) to form a chip after the probe is fixed.
  • the substrate is a substrate, such as a commercially available amino glass slide.
  • the multi-chip base substrate has an isolation structure.
  • the substrate includes a base and an isolation structure.
  • the chip-based cell forms a reactor after the probe is fixed.
  • the wafer base forms a multi-reactor chip.
  • the substrate is a solid-phase carrier used to fix probes and other auxiliary agents (if any). The surface chemical and optical properties are important factors affecting chip performance and cost.
  • chip signal detection instrument is an important condition for chip applications.
  • the most commonly used chip signal detection instruments are scanners, including confocal scanners, CCD scanners, and general scanners.
  • the chip compartment in the scanner is the place where the chip is placed during scanning, and its optical properties affect the scanning results.
  • the existing chip detection methods include two types of light detection methods and non-light detection methods.
  • non-luminous detection methods are the SELDI-TOF-MS method (Surface-Enhanced Laser Dissociation and Laser Ionization Time-of-Flight Mass Spectrometry, Surface-Enhanced Laser Desorption / Ionization-Time of Flight-Mass Spectra), such as Ciphergen, USA It includes the MetalCM-based ProteinCMp Array system.
  • Luminescence detection methods mainly include fluorescence detection method, chemical luminescence detection method and composite light irradiation detection method.
  • the current chip detection methods are based on the minimization of background signals.
  • the substrates used in fluorescence detection methods are basically transparent glass slides (such as amination, aldolization, and polylysine film substrates), and the substrates used in chemiluminescence detection methods are basically transparent glass slides, Plastic plate or metal film (such as silver film), polymer film (such as PVDF film, nylon film, nitrocellulose film, acetate fiber film, etc.) that is basically diffusible in the substrate used in the composite light irradiation detection method.
  • Plastic plate or metal film such as silver film
  • polymer film such as PVDF film, nylon film, nitrocellulose film, acetate fiber film, etc.
  • the current composite light irradiation detection method based on a diffusible polymer film base has low sensitivity; the current chemiluminescence detection method is not very sensitive; the current fluorescence detection method, although the sensitivity is higher than the previous two High, but the background noise of the slide substrate is not low, the cost of the activated substrate is high, the detection equipment is expensive and other deficiencies, which affect the large-scale application of the biochip method.
  • the reduction of its background noise is always limited and sometimes even full of contradictions.
  • the detection sensitivity of the chip is necessary to increase the activity and / or concentration of the surface-active groups on the substrate, but at the same time increase the non-specific binding activity on the substrate except for the probe points, thereby increasing the background noise Increased risk.
  • a laser excitation light source with a larger power is required in the laser confocal scanner, but it increases the cost and increases the risk of photobleaching.
  • a higher-power excitation light source is required in the CCD scanner, but it increases both the cost and the instrument noise (such as dark current).
  • a chip substrate which includes a substrate and an optional isolation structure
  • the substrate includes a substrate and optionally one or more of the following composite structures: coating, Coatings, films, and flakes, the matrix and / or at least one of the composite structures contain a colorant and / or a capture agent that directly or indirectly captures the colorant.
  • a chip which includes a reaction system and an optional labeling system.
  • the reaction system includes a substrate as described above and a probe fixed in a substrate substrate pool. .
  • the present invention provides a chip signal detection instrument, wherein at least the surface of a portion directly below the chip during scanning is an ultra-black surface having an absorbance greater than 95%, preferably greater than 98%.
  • the ultra-black surface contains an ultra-black metal salt and / or an ultra-black metal oxide.
  • a method for preparing a chip substrate according to the present invention which comprises introducing on the substrate a light-emitting substance and / or a dye and / or a coloring pigment and / Or matting agents in one or more of the following structures: coatings, coatings, films and flakes.
  • a method for preparing a chip and a chip manufactured according to the method includes mixing a probe with a dye, a color pigment, and / or a coating material containing the color pigment, and then spotting the sample into a film base pool.
  • the present invention provides a chip detection method, in which at least one substrate and / or at least one probe point of a reaction system of a used chip contains dyes and / or colored pigments that are not used as probes and / Or a paint containing a coloring pigment, the dye is selected from black, green, blue, purple, and red dyes including amino black, Coomassie brilliant blue, crystal violet, and Ponceau red; the coloring pigment is selected from Said one or more pigments: luminescent pigments including fluorescent substances, chemiluminescent pigments and electroluminescent pigments, black pigments including carbon black, metal salts, white pigments including titanium dioxide, and Yellow pigment, red pigment, blue pigment, green pigment, metallic pigment; the paint is selected from black paint, white paint, red paint, yellow paint, green paint, blue paint, indigo paint or purple paint Painted inside. detailed description
  • analysis chip or "chip” in the present invention includes but is not limited to
  • Biocliip "Microarray”, and “Bioarray” are a kind of detection device in the specified and / or quantitative analysis.
  • the specific reaction between the trace probe in the reactor and the target molecule in the sample can be addressed with an addressable address. Way to identify.
  • the core of the chip is the reactor therein, and the core of the reactor is the chip substrate and the probe fixed on the chip substrate.
  • the chip includes a microchannel chip (equivalent to in English) and a microarray chip (equivalent to "Biochip”, “Microarray”, “Bioarray” in English), but it is well known that it does not include existing rapid test reagent strips.
  • the chip of the present invention contains a single reactor or multiple reactors with or without a labeling system.
  • the distribution density of the probes on the substrate in the reactor is greater than 10 points W, and the preferred solution is greater than
  • detection device is a device used in a specified and / or quantitative analysis method, and includes a chip, a signal detection device, and the like.
  • reactor in the present invention refers to the place where the probe specifically reacts with the target and other related structures communicating with it, such as the reaction cell and Related isolation structures and inlet and outlet fluid structures.
  • substrate in the present invention refers to a product based on a substrate, optionally combined with other structures (such as an isolation structure), and used to form a chip after the probe is fixed.
  • Monolithic substrates usually do not have an isolation structure on them.
  • the substrate is a substrate (such as a commercially available amino slide).
  • the substrate includes a base and an isolation structure.
  • the chip-based cell forms a reactor after the probe is fixed, and the multi-chip base forms a multi-reactor chip.
  • the substrate is a solid support used to hold probes and other additives (if any). The surface chemical and optical properties are important factors affecting chip performance and cost.
  • film base pool in the present invention refers to a structure formed by a film base and its isolation structure.
  • probe in the present invention refers to a substance, such as an antigen, an antibody, a nucleic acid, and the like, which is immobilized on a solid-phase carrier to identify a target substance in a sample.
  • the term "contrast maximization" in the present invention The signal contrast between the target and the background is a sign of detectability.
  • C used the contrast ratio radiation to represent the detectability of a thermal imaging camera:
  • Eo is the target specific emissivity
  • Eb is the background specific radiance
  • Eo and Eb are directly proportional to the absorption of the target material and the background material, respectively.
  • C is in the best state of stealth when it approaches zero.
  • the contrast-maximizing structure refers to a structure that maximizes the contrast between the target and the background.
  • background signal enhancement in the present invention means that the background detection signal is made higher than the signal with the weakest target detection signal.
  • Attenuation of the target signal means that the weakest target detection signal is made lower than the background detection signal.
  • achromatic color in the present invention: when white light is irradiated on an object, if the reflectance of visible light of all wavelengths is greater than 80%, the object is white; if the reflectance of visible light of all wavelengths is less than 7%, the object is black; If the reflectance of visible light is uniformly greater than 7% and less than 80%, the object is gray; a type of color composed of white, gray, and black is called an achromatic color in the present invention.
  • coating in the present invention refers to immobilizing a substance in a solid state on a solid support, For example, coating points, coating surfaces, etc., on which the dye molecules are fixed on the glass slide by ion adsorption, affinity adsorption, and the like.
  • coating in the present invention refers to a dry film having a thickness of less than 1 mm formed by coating a solid phase material with a coating material.
  • film in the present invention refers to a non-porous or perforated planar material having a thickness of less than 0.3 mm.
  • sheet in the present invention refers to a non-porous or perforated planar material having a thickness of 0.3 mm or more.
  • labeling substance in the present invention refers to a substance used to form or participate in the formation of a detection signal, such as fluorescein, a labeling substance commonly used in chip detection.
  • ligand in the present invention refers to a substance used to capture its ligand (Ligate) through affinity, such as one or more of the following: antigen, antibody, ligand, ligand, polypeptide And single-stranded or multi-stranded DNA, RNA, nucleotides.
  • coloring in the present invention refers to making the preparation have certain properties of light absorption, reflection, refraction and the like so as to form optimized hue and / or lightness and / or saturation in visible light or under a selected wavelength of light.
  • colorant refers to a substance that can color a preparation.
  • coloring pigment in the present invention refers to a substance that has no affinity for the coloring object and is mainly colored by combining other film-forming materials such as resin and adhesive with the coloring object, such as carbon black, mica titanium pearlescent coloring pigment, and azo coloring pigment. , Phthalocyanine color pigment, colorless fluorescent color pigment.
  • the coloring pigments in the present invention do not include extender pigments according to the custom of the coating industry.
  • the extender pigments generally refer to bulk fillers or substances without coloring functions. Examples of extender pigment colors can be the metallic silver color of a silver foil base, and a film of a base film. Essence and so on.
  • die in the present invention refers to a substance that has an affinity for a colored object and can color the colored object.
  • paint in the present invention refers to a substance applied to a substrate of a film to obtain a specific function of a dry film having a thickness of less than 1 mm.
  • Many paints contain colored pigments, such as various paints and various inks.
  • target in the present invention refers to the entirety of all substances related to the detection signal at the probe point in the chip pool when the signal is detected, including the probe, the target (if any) captured by the probe, and the marker ( If so, in addition to the probe, a probe carrier (for example, a nanocarrier) and a dye, a coloring pigment, a marker, and the like of the present invention may be provided on the probe point.
  • a probe carrier for example, a nanocarrier
  • film background in the present invention refers to the entirety of all substances related to the detection signal in the detection area except the target in the film base pool when the signal is detected.
  • the present invention aims to provide more alternative substrates / chips, and / or improve chip detection sensitivity.
  • one of the prerequisites for providing more alternative substrates / chips is that they need to have a sufficiently high detection sensitivity.
  • the objectives of the present invention are related to the objectives of current stealth technology. For example, for infrared stealth technology, Maclean et al. Used the contrast ratio radiation C to represent the detectability of the thermal imager: C2 Eo-Eb. Where Eo is the specific emissivity of the target, Eb is the specific emissivity of the background, and Eo and Eb are directly proportional to the absorption of the target and background materials, respectively.
  • Stealth technology is devoted to minimizing the contrast between the target and the background.
  • the technology of the present invention is devoted to maximizing the contrast between the target and the background, thereby improving the chip detection sensitivity and / or providing more options with a sufficiently high detection sensitivity Substrate / chip.
  • the technique of the present invention may be referred to as a manifestation technique, and the detection method of the present invention may be referred to as a manifestation technique detection method.
  • by increasing the background luminous ability of the chip substrate instead of reducing the current background luminous ability we were surprised to find that not only did the detection sensitivity not decrease, but it was significantly improved.
  • the research of the present invention selects "revealing body” as the technical basis, and seeks to minimize the background noise of the chip base and seek the signal of the weakest detection target (usually a negative detection target) of the chip base background signal.
  • the maximization of the difference ' 5 and "the maximum optical contrast between the labeling substance and the substrate to ensure or improve the detection sensitivity" two technical routes to achieve this goal.
  • the roughness Ra of the chip substrate surface of the present invention is between 0.02 and 5.0 ⁇ m, and preferably between 0.25 and 5.0 ⁇ m.
  • the chip substrate of the present invention includes a substrate and an optional isolation structure.
  • the substrate includes a substrate and optionally one or more of the following composite structures: a coating, a coating, a film, and a sheet.
  • the substrate And / or at least one of the composite structures contains a colorant and / or a capture agent that directly or indirectly captures the colorant. It should be noted here that the capture agent that captures the colorant directly or indirectly is not used to capture a probe.
  • the current chip substrates are all transparent or uncolored substrates.
  • the uncolored film base only retains its base color (such as the color of extender pigments) without adding dyes and coloring pigments. Examples thereof may be the metallic silver color of silver foil base, the film color of thin film base, and the like.
  • a coating, a coating, a film, and a wafer containing a light-signal-related substance In the front and / or back of the substrate and / or sandwich.
  • a coating containing a fluorescent substance in the substrate Table 1
  • the surface of the substrate contains a coating containing a luminescent pigment
  • the surface of the substrate contains a reflective film
  • the surface of the substrate contains a flake containing a luminescent pigment, and the like.
  • the colorant and / or capture agent is formed or involved in forming a colored background and / or a signal weakening target at least under signal detection conditions when the substrate is applied, and the colored background includes One or more of the following backgrounds: a luminous background that emits signal light, a reflective background with a signal light reflectance greater than 50%, an achromatic color background, and a colored background, the signal weakens the target, and its detection signal is related to the optical signal The presence of matter weakens.
  • Example 1 an example of a substrate having a light-emitting background (for example, a substrate having a luminescent pigment film on the surface of the substrate) and a light-reflecting background (for example, a substrate having a reflective film on the surface of the substrate) is given.
  • a substrate having a luminescent background and / or a reflective background is also referred to as a background signal enhancement substrate in the present invention.
  • an example of a substrate having an achromatic color background black substrate, white substrate
  • a color background blue substrate
  • a substrate having an achromatic color background and / or a colored background is also referred to as a color substrate in the present invention.
  • the signal attenuation target and / or colored background preferably a luminescent background and / or a reflective background participate in forming a high noise signal ratio
  • the high noise signal ratio refers to a background signal ratio to a negative target Or a blank control target or an extreme weak positive target has a high detection signal, preferably at least 50% higher, more preferably at least 300% higher.
  • Increasing the noise-to-signal ratio as a method for improving sensitivity is an important point of the present invention.
  • the enhanced background of the substrate can be enhanced by one or more of the following methods: adding a luminescent pigment (such as fluorescein) to the substrate, and adding a coating or coating containing a luminescent pigment or reflection on the front and / or back of the substrate (Such as a luminescent coating on the surface and / or back of the substrate on which the probe is fixed), a thin film (such as a luminescent film or reflective film on the surface and / or the back of the substrate), with or without detection targets Porous flakes (such as luminescent flakes covering the surface and / or back of the substrate), and active groups derived from the surface of the substrate that can directly or indirectly bind luminescent pigments (such as coating protein A on the substrate and then Binding of rhodamine-labeled IgG-example tablet before or after detection reaction).
  • a luminescent pigment such as fluorescein
  • Signal weakening targets can be achieved by adding dyes, coloring pigments and / or coatings that reduce signal light emission and / or reflection or increase signal light absorption at the quasi-fixed probe points, and / or add Coatings, coatings (eg, matt coatings on the surface and / or back of the substrate) of these dyes, pigments and / or coatings, films (eg, covering the surface of the substrate on which the probe is fixed and / or The light-reducing film on the back surface), and / or the sheet (for example, a light-reducing sheet containing a test substance colorant covering the surface of the substrate and / or the back surface).
  • Coatings coatings (eg, matt coatings on the surface and / or back of the substrate) of these dyes, pigments and / or coatings, films (eg, covering the surface of the substrate on which the probe is fixed and / or The light-reducing film on the back surface), and / or the sheet (for example, a light-reducing sheet containing a test substance
  • the colored background preferably an achromatic color background and / or a colored background participates in forming a high chromatic aberration ratio
  • the high chromatic aberration ratio refers to the entire wavelength or a part of the wavelength between the background and the target.
  • the absolute value of the signal light or the absorbance difference in reflectance of not less than 50%, preferably not less than 70% 0 of the higher ratio means a significant color difference between background and object in hue, lightness and saturation, or
  • the absolute value of the difference between the absorptance or reflectance of the signal light of all or part of the wavelength between the background and the target is not less than 50%, preferably not less than 70%.
  • the color difference ratio as a method of improving sensitivity, which is another focus of the present invention.
  • the difference between the selective absorption rate of the signal light between the background and the target for example, the achromatic color difference between a black background with an absorption rate greater than 95% and a light-emitting target containing a fluorescent substance (the absorption rate is set to 0 in the present invention)
  • the maximization of the value is conducive to the target's self-explanation and thus improves the sensitivity.
  • the difference between the selective reflectivity of the signal light between the background and the target is also conducive to the target's appearance and thus improves sensitivity.
  • the achromatic color background and / or color The background is also called a color background.
  • Obtaining a color background includes adding dyes and / or coloring pigments and / or coatings containing coloring pigments to a substrate, or adding dyes and / or coloring pigments and / or coloring pigments to a base.
  • One or more of the following structures of a coating coating, coating, film, and sheet.
  • the substrate is selected from the group consisting of modified or unmodified glass, plastic, and metal.
  • the derivative activation group of the modified glass slide may include one or a combination of any two or more of the derivative activation groups of the existing glass chip substrate: amino, epoxy, aldehyde, and hydrazide (-CO-NHN3 ⁇ 4), aminoureido (3 ⁇ 4N-NH-CONH-), diethylaminoethyl (DEAE), diethylmono (2-hydroxypropyl) aminoethyl (QAE), carboxymethyl ( CM), sulfopropyl (SP), mercaptoethylpyridine (MEP), siloxane group, thiol group.
  • the colorant includes one or more of the following substances: a light-emitting substance, a dye, a coloring pigment, and a matting agent.
  • Luminescent substances, dyes, coloring pigments, and matting agents are a well-known concept in the coloring of materials such as plastics and coatings, and have certain contents.
  • the luminescent substance is selected from one or more of the following substances: rhodamine, CY3, CY5, Alexa, seaweed protein, rare earth compound-based fluorescent substances, chemiluminescent substances, and electrochemical light-emitting substances.
  • the dyes include amino black, Coomassie brilliant blue, crystal violet, Ponceau red, printed paint paddles (7701 FBRN BLACK FBRN, 6201 Scarlet FGG SCARLET FGG, 6101 F7G BRILLIANT YELLOW F7G), water-based vapor dyes (water-based blue, Water-based green, water-based white) including black, purple, green, blue, indigo water-based dyes, water and oil amphoteric dyes.
  • the coloring pigment is selected from one or more of the following pigments: black pigments including carbon black, metal salts, white pigments including titanium dioxide, and yellow pigments, red pigments, blue pigments, green pigments, Color pigments including metallic pigments.
  • the fluorescent coloring pigment includes one or more of the following substances: rhodamine, CY3, CY5.
  • the dyes are black, purple, green, blue or indigo water-based dyes including amino black, Coomassie brilliant blue, crystal violet, water-oil amphoteric dyes, printing paint pastes, and the like.
  • the roughness Ra of the surface of the substrate is between 0.02 and 5.0 m, preferably between 0.25 and 5.0 m.
  • the paint of the coating layer is selected from black, white, and various colored paints and / or inks.
  • the colors include red, yellow, green, blue, cyan, and purple colors.
  • both the ink and the paint have the property of coloring the substrate.
  • the substrate containing the colorant in the substrate and / or the composite structure contains a substance capable of binding a probe.
  • the probe-binding substance includes one or more of the following known probe-binding organic substances and derivatives thereof: nitrocellulose, polystyrene, polyvinyl chloride, amino resin, polysaccharide, polyamino acid, Polyacrylate, polysulfone, polyethersulfone, etc.
  • the color paint of the coating layer includes one or more of the following paints: pearl black, magic black, pearl white, pearl blue, matte black, claret, scarlet, medium blue, etc. .
  • the capture agent includes one or more of the following ligands: protein A, protein G, biotin, avidin, antigen, antibody, anti-antibody, polypeptide, DNA, etc. .
  • an antibody may capture an anti- antibody that binds to rhodamine to indirectly capture rhodamine, and so on.
  • the substrate containing the optical signal related substance in the substrate and / or the composite structure contains a substance capable of binding a probe.
  • the probe-binding substance includes one or more of the following organic substances and derivatives thereof: nitrocellulose, polystyrene, polyvinyl chloride, amino resin, polysaccharide, polyamino acid, polyacrylate, polysulfone, Polyethersulfone, etc.
  • the present invention also provides a chip including a reaction system and an optional marking system, the reaction system including the substrate according to the present invention as described above and a probe fixed in a substrate substrate pool. needle.
  • the chip of the present invention can detect a signal for detecting a reaction result by one of the following instruments: a confocal scanner, a CCD scanner, a visible light scanner, and the like.
  • the substrate of the substrate and the marking system are one of the following combinations:
  • the substrate is red, orange, yellow, green, blue, cyan or purple under white light, preferably black
  • the marking substance is a luminescent substance or a white, light red, light orange, light yellow, light green, light blue, light cyan, or light purple substance
  • the substrate is light red, light orange, light yellow, light green under white light
  • the marking substance is red, orange, yellow, green, blue, cyan or purple, and preferably, the black substance.
  • the probe is fixed by being mixed with nano particles and fixed in the substrate, wherein the average particle diameter of the nano particles is 1 to 500 nm.
  • the marking system contains a dye.
  • the dye is a black, purple, green, blue or indigo dye including amino black, Coomassie brilliant blue, crystal violet, and the like.
  • the present invention provides a chip signal detection instrument, in which at least the surface of the portion directly below the chip during scanning is an ultra-black surface with an absorbance greater than 95%, preferably greater than 98%.
  • the ultra-black surface contains an ultra-black metal salt and / or an ultra-black metal oxide, and the like.
  • the present invention provides a method for preparing a chip substrate, which includes introducing one or more of the following structures containing a luminescent substance and / or a dye and / or a coloring pigment and / or a matting agent on the substrate: Coatings, coatings, films and sheets.
  • the present invention also provides a method for preparing a chip, which comprises mixing a probe with a dye, a coloring pigment, and / or a coating material containing the coloring pigment, and then spotting the sample into the film base pool.
  • the dyes are black or colored water-based dyes including amino black, Coomassie brilliant blue, crystal violet, etc., water and oil amphoteric dyes, or printing paint paste.
  • the coloring pigment is selected from one or more of the following pigments: black pigments including carbon black, metal salts, etc., and white pigments including titanium dioxide And pigments, including yellow, red, blue, green, and metallic pigments.
  • the paint is selected from black, white and various colored paints and / or inks.
  • the coating contains a substance that can bind a probe.
  • the probe-binding substance includes one or more of the following organic substances and derivatives thereof: nitrocellulose, polystyrene, polyacrylate, polysulfone, polyethersulfone, polyvinyl chloride, amino resin, polysaccharide , Polyamino acids, etc.
  • the paint color paint include one or more of the following paints: pearl black, magic black, pearl white, pearl blue, matte black, scarlet, medium blue, and the like.
  • the present invention provides a chip in which at least one probe point in a reaction system contains the dye and / or the colored pigment and / or the coating ⁇ "that is not a probe.
  • the present invention also provides a chip detection method, in which at least one substrate and / or at least one probe point of the reaction system of the used chip contains dyes and / or colored pigments and / or colored pigments that are not used as probes.
  • the paint is selected from black, green, blue and indigo dyes including amino black, Coomassie brilliant blue, crystal violet;
  • the coloring pigment is selected from one or more of the following pigments: including Light-emitting pigments including fluorescent substances, chemiluminescent pigments and electrochemical light-emitting pigments, black pigments including carbon black, metal salts, etc., white pigments including titanium dioxide, and yellow pigments, red pigments, A blue pigment, a green pigment, a metallic pigment, and the like;
  • the paint is selected from a colored paint including black paint, white paint, red paint, yellow paint, green paint, blue paint, indigo paint, or purple paint.
  • the advantages of the substrate according to the present invention are that it can impart a high detection sensitivity to the chip of the final product, or a high degree of freedom of selection, a low cost, etc., while it can have a sufficiently high detection sensitivity.
  • the advantages of the chip of the present invention are high detection sensitivity, or a high degree of freedom of selection, low cost, etc. while a sufficiently high detection sensitivity can be achieved.
  • the chip signal detection instrument of the invention has the advantage of high detection sensitivity.
  • the advantages of the chip detection method of the present invention are high detection sensitivity, or a high degree of freedom of selection, low cost, etc., while a sufficiently high detection sensitivity can be achieved.
  • Embodiment of the present invention give examples of implementation principles of the present invention, and cannot be understood that the present invention is limited to these embodiments.
  • the preparation of a plurality of substrate pool substrates wherein the isolation structure of the substrate pool is coated on the above glass slide with a highly hydrophobic organic silicon coating (Chengdu Chenguang Chemical Design Institute), and dried to form a film (film thickness less than 0.05 mm) was formed on the substrate (refer to our other invention: " ⁇ a biochip with a minimum height of the reactor isolation structure and a preparation method", patent application number 03117397.7).
  • Eight substrate cells are formed for each substrate. The size of each substrate cell is 4.5 mm X 4.5 mm, and the width of the isolation structure between the substrate cells is 4.5 mm.
  • the probes used in this example are HCV antigen (Institute of Liver Diseases, Beijing People's Hospital, China) and HIV 1 + 2 antigen (Institute of Liver Diseases, Beijing People's Hospital, China).
  • HCV antigen Institute of Liver Diseases, Beijing People's Hospital, China
  • HIV 1 + 2 antigen Institute of Liver Diseases, Beijing People's Hospital, China
  • two antigens each have three points with a diameter of 200 ⁇ m, and the distance between them is 800 m, forming a 2 ⁇ 3 array.
  • the probe density on the reactor substrate was greater than 96 points / cm 2 .
  • sample 1 is HCV antibody positive serum
  • sample 2 is HIV 1 + 2 antibody positive human serum
  • sample 3 is a positive control (a mixture of HCV antibody and HIV 1 + 2 antibody positive serum control )
  • Sample 4 is the negative control (HCV antibody and HIV 1 + 2 antibody Are negative serum controls). All samples were pre-tested using the classic ELISA method under serum '20-fold dilution reaction conditions.
  • the background signal enhancement substrate of this embodiment includes a substrate having a luminescent background and a substrate having a reflective background.
  • the reference substrate used in this embodiment is the amino slide in Table 1, and the prepared substrates are listed in Table 2. Background signal enhancement substrate
  • Substrate background signal enhancement structure Substrate detection signal Noise ratio ***
  • noise signal ratio [(background signal-negative signal) I negative signal] absolute value of the substrate 1 prepared in this embodiment, which is a polystyrene substrate with a fluorescent substance added thereto.
  • the method for preparing a substrate includes adding a fluorescent pigment ZnS-Mn to a thermoplastic polystyrene used for preparing an enzyme-labeled plate, and then obtaining the size by molding, and the size is 75x25x1 mm.
  • the method for preparing the multi-chip base is as described above.
  • the film base 2 prepared in this embodiment is a polyvinyl chloride film-glass slide composite film base to which a fluorescent substance is added.
  • the method for preparing the substrate includes thermally bonding a polyvinyl chloride film that reflects light at a wavelength of 532 nm to a glass slide described in Table 1.
  • the method for preparing the multi-chip base is as described above.
  • the base 3 prepared in this embodiment is a protein A-coated base.
  • the preparation method includes coating protein A (Shanghai Biological Products Research Institute) on the amino slides described in Table 1 by using a general protein coating technology.
  • the base 4 prepared in this example is a rhodamine-polypeptide coating substrate.
  • the preparation method includes coating the formed rhodamine-polypeptide on a semicarbazide glass slide described in Table 1 by using a known rhodamine-polypeptide technology.
  • the polypeptide used is Epstein-Barr virus, VCA antigen fragment.
  • the film base 5 prepared in this embodiment is a film base with a fluorescent substance-containing coating on the front side, and the preparation method includes a rhodamine-white paint light-emitting film base on the front side.
  • the thickness of the coating formed is around 30 ⁇ .
  • the film base 6 prepared in this embodiment is a film base with a fluorescent substance-containing coating on the back surface, and the preparation method includes a rhodamine-white paint light-emitting film base on the back surface.
  • the thickness of the coating formed is around 30 ⁇ m.
  • the film base 7 prepared in this embodiment is a front hole light-emitting film base.
  • the preparation method includes leaving holes in a place where a polyvinyl chloride film has a reaction cell, and bonding the top surface of the aldehyde-based glass slide by a thermal bonding process. .
  • the light-emitting film is a polyvinyl chloride film.
  • the film base 8 prepared in this embodiment is a light-emitting film base with a hole on the back.
  • the preparation method includes leaving holes in the place where the polyvinyl chloride film has a reaction cell, and bonding the bottom surface of the PVP-coated glass through a thermal bonding process.
  • the light-emitting film is a polyvinyl chloride film.
  • a preparation method includes combining a non-porous fluorescent film on the back of the amino dolphin slide with a thermal bonding process.
  • the slide glass is the amino slide glass described in Table 1.
  • the substrate 10 prepared in this embodiment is a fluorescent substrate with holes on the back.
  • the preparation method includes using an amino slide as described in Table 1 as the substrate, and spotting and coating by conventional methods.
  • the perforated fluorescent sheet is a sheet of PVC compression molding containing a fluorescent substance, which is punched and cut into a sheet with a plane size of 75x25 mm (thickness of 80 ⁇ m, pore diameter of 200 ⁇ m, and the distribution of holes corresponds to spotting. Distribution of sample points).
  • the prepared substrates were formed from the eight substrate pool substrates formed by the aforementioned multi-piece substrate formation method.
  • the substrates 0 to 10 in Table 2 were prepared from the substrates 0 to 10. ⁇
  • the prepared substrate was determined to have an enhanced background signal as follows: A HCV antigen and HIV 1 + 2 antigen mixture (2 mg / ml each) was spotted into a substrate pool on the substrate according to a known chip spotting method. The 4 points in the pool form a 2X2 square matrix with a diameter of 200 ⁇ m and a pitch of 800 m. Then take the No. 4 sample (serum control with negative HCV antibody and HIV 1 + 2 antibody) as the target sample, rhodamine-labeled goat anti-human anti-antibody (Jackson ImmunoRresearch Laboratories, USA) as the label, and according to the known chip
  • the detection method detected negative targets and substrate background signals (Table 2).
  • the background signal enhancement chip of this embodiment is formed by fixing a probe in the substrate base pool of the above background signal enhancement substrate (chip 0-6 in Table 3), or after fixing the probe on an amino slide, the substrate is processed. Coating of capture agents or substances containing luminescent substances (chips 7 and 8 in Table 3).
  • the method of fixing the probe is as follows: In each of the above-mentioned prepared substrate pools, add HCV antigen (1.5 mg / ml) and HIV1 + 2 antigen solution (1.5 mg / ml) to the nanoparticles (silica oxide nanoparticles (SP1) ), 15-25 legs, Zhejiang Zhoushan Mingri Nano Materials Co., Ltd.), and then spotted in the substrate pool to form a reactor.
  • the coating method of the capture agent or the luminescent substance-containing substance is: The capture agent or the luminescent substance-containing substance at a concentration of about 0.1 mg / ml is performed according to a well-known chip blocking method.
  • the prepared chips are listed in Table 3. Background signal enhancement chip
  • the background signal enhancement structure of the prepared chip was determined as follows: A mixture of human HCV antibody negative serum and human HIV antibody negative serum (1: 1) was used as a target sample, and rhodamine-labeled goat anti-human anti-antibody (Jackson ImmunoRresearcli Laboratories, USA) ) Is a marker, and a negative target and a substrate-back signal are detected according to a known chip detection method (Table 3). Background signal enhancement detection method
  • the reference picture used is chip 0 in Table 3.
  • the method for preparing a substrate in this embodiment is made by spotting a dye or coating with a spotter to multiple base cells, or forming a target signal weakening point on the back of a glass probe to which a probe is to be fixed.
  • a target signal weakening point can also be formed by spotting a dye or coating with a spotter to or unactivated the point where the probe is to be fixed on the slide or the back of the point where the probe is to be fixed on the slide. Then, according to the aforementioned method for forming a multi-chip base cell, a multi-chip base cell substrate is prepared.
  • the substrates prepared in this example are listed in Table 5.
  • the preparation of the substrate bl is as follows: Using a spotter, the crystal violet (company) solution with a concentration of 2 mg / ml is distributed according to the probe point to the 8-piece base cell glass prepared according to the aforementioned multi-chip base cell preparation method On a chip (2 X 3 array, spot diameter 200 ⁇ m), react at room temperature for 1 hour, then wash and dry.
  • the preparation of the substrate b2 is as follows: Using a spotter, a Coomassie brilliant blue (company) solution with a concentration of 2 mg / ml is spotted onto a pseudo-point probe on 8 base-cell amino slides prepared according to the aforementioned multi-chip base-cell preparation method The back of the spot (2 X 3 array, dot diameter 200 m) was washed at room temperature for 1 hour and then dried.
  • the substrate b3 was prepared as follows: Using a spotter, pearl black paint (Shanghai Qifu Industrial Development Co., Ltd.) was distributed according to the probe point distribution points to the 8-piece base-loaded glass prepared by the above-mentioned multi-chip base-cell preparation methods. On-chip (2 ⁇ 3 array, spot diameter 200 ⁇ m), and then dried.
  • the substrate b4 was prepared as follows: Using a spotter, pearl black paint (Shanghai Qifu Industrial Development Co., Ltd.) was spotted onto the quasi-point probe points on the 8 base cell amino slides prepared according to the aforementioned multi-chip base cell preparation method. Back side (2 X 3 array, spot diameter 200 ⁇ m), and then dried. Table 5: Target signal attenuation substrate
  • Noise signal ratio [(background signal-negative signal) I negative signal] absolute value of the target signal of the substrate prepared in this example is determined as follows: HCV antigen and HIV 1 + 2 antigen are determined according to the known chip spotting method The mixture (2 mg / ml each) was spotted into the substrate pool on the substrate, and 4 spots were formed in each substrate pool to form a 2X 2 square matrix with a diameter of 200 um and a pitch of 800 ⁇ m. Then take the No.
  • chip 0 is a comparative chip using 8 base pool amino slides as the base.
  • the chips bl, b2, b3, and b4 in this embodiment are prepared according to a known spotting method.
  • HCV antigen 1.5 mg / ml
  • HIV 1 + 2 antigen 1.5 mg / ml
  • the antigen and HIV 1 + 2 antigen dots were made by overlapping the target signal weakening dots (2 X 3 array, dot diameter 200 ⁇ m) (Table 6).
  • Table 6 Target signal attenuation chip
  • the target signal attenuation structure of the chip prepared in this embodiment is determined as follows: A mixture of human HCV antibody negative serum and human HIV 1 + 2 antibody negative serum (1: 1) is used as a target sample, and rhodamine-labeled goat anti-human anti-antibody (Jackson ImmunoRresearch Laboratories, USA) As a marker, a negative target and a substrate-based background signal were detected according to a known chip detection method (Table 6).
  • Example 2 the same four samples as in Example 1 were added to the reactor of the chip.
  • Each of the 4 reactors of each biochip was filled with a 1: 500 diluted sample. The sample volume was 15 ⁇ 1. After 30 minutes of reaction, the product was washed five times, and the washing solution was added in an amount of 25 ⁇ 1 each time. The amount of the labeled substance was 15 ul, washed 5 times after the reaction, and the washing solution was added 25 ⁇ each time. After drying, scanning was performed.
  • the scanner is a confocal laser scanner (Afymetrix's GMS 418 chip scanner). The scanning light wavelength is 532 nm and the emission light wavelength is 570 nm.
  • the read signal is processed by the processing software (ZoCSoft ImageBoost), and the average value is obtained. The results are shown in Table 7. Table 7: Target signal attenuation chip detection results
  • the background signal enhancement / target signal attenuation substrate prepared in this example is shown in Table 8.
  • the method for preparing the substrate cl-c5 includes the background signal enhancement and the target signal attenuation: (1) The method for background signal enhancement is the same as the method for preparing the background signal enhancement substrate in Example 1 (the substrates cl, c2, c3, c4, c5 corresponds to the substrates 3, 4, 5, 6, 9 respectively; (2) The method for weakening the target signal is the same as the method for preparing the target signal attenuating substrate in Example 2. The pearl black paint (Shanghai Qifu) Industrial Development Co., Ltd.) Press the distribution points of the probe points to the front of the substrate base and dry. Clothing 0: Header Jj ' ⁇ Signal enhancement / Target signal weakening substrate Substrate Background signal enhancement Target signal weakening Detection signal Noise to signal ratio Negative
  • Substrate c3 contains fluorescent substance on the front side and contains target letter on the front side 12783 -7599 2.7 Base point of coated film
  • Substrate c4 contains fluorescent substance on the back 11456 -6973 on the front 2.6 The weakening point of the base of the coated film
  • Substrate c5 has a reflective film on the front with target letter 10591 -7613 2.4 weakening point on the front
  • the determination of the noise-to-signal ratio of the substrate prepared in this embodiment is as follows: A HCV antigen and an HIV antigen mixture (2 mg / ml each) are spotted into a substrate pool on the substrate according to a known chip spotting method. There are 4 points in the pool to form a 2 ⁇ 2 square matrix with a diameter of 200 ⁇ ⁇ and a pitch of 800 ⁇ ⁇ . Then take the No.
  • the difference between the background signal enhancement / target signal attenuation substrate background and the negative signal is greater than the difference between the background signal enhancement substrate or the target signal attenuation substrate background and the negative signal.
  • Chips cl, c2, c3, c4, and c5 were prepared by spotting the HCV antigen (1.5 mg / ml) and HIV 1 + 2 antigen (2 mg / ml) according to the well-known spotting method.
  • the HCV antigen and HIV 1 + 2 antigen spots are coincident with the target signal attenuation structure (2X 3 array, dot diameter 200 ⁇ m) and Made (Table 9).
  • the determination of the background signal enhancement / target signal attenuation is as described above. Scene signal increase / target signal decrease. Background signal enhancement / target signal attenuation detection method
  • Example 2 the same four kinds of samples as in Example 1 were respectively added to the reactor of the chip.
  • Each of the 4 reactors of each biochip was filled with a 1:50 diluted sample.
  • the sample volume was 15 ⁇ 1.
  • the labeling amount was 15 ⁇ 1, and the reaction solution was washed twice.
  • the washing solution was added 15 ⁇ 1 each time.
  • scanning was performed.
  • the scanner is a confocal laser scanner (Afymetrix's GMS 418 chip scanner).
  • the scanning light wavelength is 532 nm and the emission light wavelength is 570 nm.
  • the read signal is processed by the processing software (ZoCSoft lmageBoost), and the average value is obtained.
  • Table 10 Background signal enhancement / target signal attenuation chip detection results
  • the colored background substrates prepared in this example are listed in Table 11.
  • a base is prepared, and the obtained base is then an 8-base base substrate formed by the aforementioned multi-base base formation method.
  • the film base dl prepared in this embodiment is a black plastic film base, and the colorant is channel black, which is a thermoplastic polystyrene used for the preparation of microplates. After adding a filler such as channel black, it is molded. Made by molding, with dimensions of 75x25x1 mm.
  • Film bases d2, d3, and d4 are front-side colored coating film bases, and the coatings are pearl black spray paint (Shanghai Qifu Industrial Development Co., Ltd.), matte black (Shanghai Qifu Industrial Development Co., Ltd.) and In Chuanyang automatic white spray paint (Chengdu Hongguang Coating Factory), the colorants in the paint are the following pigments: trough carbon black, titanium oxide, pearl black and pearl green paints (pearlescent substances) are added as matting agents.
  • the coatings are sprayed on the top surface of the glass slide and dried to form a colored coating (thickness of about 30 ⁇ m). Their roughness Ra is between 0.4 and 0.5 ⁇ m (as measured by Chengdu Metrological Supervision and Verification Test).
  • the film base d5 is a back-colored coating film base, and the paint is pearl black (Shanghai Qifu Industrial Development Co., Ltd.).
  • the coating is sprayed on the back of the semi-urea-based glass and dried to form a colored coating (thickness around 30 ⁇ m).
  • a double-sided colored coating substrate can also be prepared.
  • Film bases d6 and d7 are front-side colored film bases.
  • the coloring films are black and white polystyrene films (about 45 m thick, self-made), and the coloring agents of the coloring films are channel black and titanium dioxide.
  • the colored film was thermally bonded to the top surface of the glass slide.
  • the film base d8 is a backing color film film base.
  • the coloring film is a black polystyrene film (about 45 ⁇ m thick, self-made), and the coloring agent of the coloring film is channel black.
  • the colored film was thermally bonded to the back of the semi-urea-based glass. Table 11: Colored background film base
  • Rhodamine-labeled goat anti-human secondary antibody was purchased from Jackson ImmunoRresearch, USA Laboratories.
  • Dye-labeled anti-antibodies of which the anti-antibodies are goat anti-human secondary antibodies (Beijing Institute of Biological Products), the dyes are amino black (Chengdu Kelong Chemical Reagent Factory), Coomassie Blue (Shanghai Boao Biotechnology Co., Ltd.) and crystals Purple (Chengdu Kelong Chemical Reagent Factory).
  • the dye was dissolved in PBS buffer to a concentration of 2 mg / ml, and then mixed with an equal volume of sheep anti-human secondary antibody at a concentration of 2 mg / ml, and reacted at room temperature for 1 hour. The free dye was then removed by purification to obtain a purified dye-labeled anti-antibody. . Optimize the preparation of contrast color chips
  • the optimized contrast color chips (Table 12) in this embodiment are all 8-reaction cell chips. It is formed by fixing the aforementioned probe in a substrate pool by a known method.
  • Table 12 Optimized Contrast Combination Chip
  • the reference photo base used was a transparent amino-modified glass slide with a size of 75x25x 1 mm (Telece International, USA, hereinafter referred to as the base 0).
  • Example 2 For chips 0, dl, d2, d3, d5, d6, and d8, the experiment is the same as in Example 1 Each sample was added to the reactor of the chip. Each of the 4 reactors of each biochip was filled with a 1: 500 diluted sample. The sample volume was 15 ⁇ 1. After 30 minutes of reaction, the product was washed five times, and the washing solution was added in an amount of 25 ⁇ 1 each time. The labeling amount was 15 ⁇ 1, and the reaction solution was washed 5 times. The washing solution was added each time at 15 ⁇ 1. After drying, scanning was performed. The scanner is a confocal laser scanner (Afymetrix's GMS 418 chip scanner). The scanning light wavelength is 532 nm and the emission light wavelength is 570 nm. The read signal is processed by the processing software (ZoCSoft ImageBoost), and the average value is obtained. Negative or positive results (Table 13).
  • the mixture After mixing the probe with the pigment or coating, the mixture is spotted on the substrate of Table 1, Table 2, Table 5, Table 8 or Table 11 by using a manual spotter to make a colored probe point chip. (Table 14).
  • the paint used in this embodiment is the same pearl black and pearl black as in Example 4, and the probes are the aforementioned HCV antigen and HIV 1 + 2 antigen (final concentrations are 1.5 mg / ml each).
  • Table 14 lists the chips prepared in this example. Table 14
  • Example 2 the same four samples as in Example 1 were added to the reactor of the chip.
  • a 1:50 diluted sample was added.
  • the sample volume was 15 ⁇ 1.
  • the product was washed five times, and the washing solution was added in an amount of 25 ⁇ 1 each time.
  • the amount of the labeled substance was 15 ⁇ .
  • washing was performed 5 times.
  • the washing solution was added each time at an amount of 15 ⁇ 1, and the scanning was performed after drying.
  • the scanner is a laser confocal scanner (Afymelxix GMS 418 chip scanner), which scans the excitation light wavelength of 532 nm and the emission light wavelength of 570 nm.
  • the read signal is processed by the processing software (ZoCSoft lmageBoost), and then averaged to obtain The results are shown in Table 15.

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Abstract

La présente invention concerne une matrice de puce de dosage, une puce comprenant la matrice au moyen de laquelle on peut analyser qualitativement et/ou quantitativement un composant d'intérêt contenu dans un échantillon, notamment dans un échantillon biologique, et un équipement pouvant s'utiliser pour mesurer la puce. La matrice de puce basée sur la présente invention est constituée d'une base et d'une structure isolante arbitraire, ladite base comprenant un substrat et une ou plusieurs structures composites sélectionnées parmi les revêtements, les couvertures, les films et les tranches. Ce substrat et la ou les structures composites peuvent comprendre une ou plusieurs substances réagissant au signal optique par l'absorption, l'émission et/ou la réflexion de signaux optiques, par exemple, des substances luminescentes, des colorants, des pigments, des agents de matage et leurs agents de capture directs ou indirects. La présente invention concerne aussi un procédé de préparation de la matrice et de la puce, ainsi que la mesure de la puce.
PCT/CN2003/001091 2003-03-13 2003-12-19 Matrice de puce, puce comprenant une matrice et leur preparation et application WO2004081570A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU2003289617A AU2003289617A1 (en) 2003-03-13 2003-12-19 A chip matrix, a chip comprising the matrix and their preparation and application
CN200480000649.8A CN1735807A (zh) 2003-12-19 2004-07-01 芯片检测方法及相关装置
PCT/CN2004/000713 WO2005059553A1 (fr) 2003-12-19 2004-07-01 Essai biologique par puce a adn et equipement correspondant
PCT/CN2004/000839 WO2005083432A1 (fr) 2003-12-19 2004-07-21 Procede et dispositif d'analyse de puce multireacteur

Applications Claiming Priority (6)

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CN 03117446 CN1250969C (zh) 2003-03-13 2003-03-13 一种对目标物进行定性和/或定量分析的检测装置及其检测方法
CN03117446.9 2003-03-13
CN03117645.3 2003-04-08
CNB031176453A CN100347545C (zh) 2003-04-08 2003-04-08 一种对样品中的目标物进行定性和/或定量分析的方法及其检测装置
CNA031177875A CN1514243A (zh) 2003-04-30 2003-04-30 对目标物进行定性和/或定量分析的方法、装置和标记物及检测试剂盒
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US5194393A (en) * 1989-11-21 1993-03-16 Bayar Aktiengesellschaft Optical biosensor and method of use
US5777372A (en) * 1995-03-01 1998-07-07 Kabushiki Kaisha Kobe Seiko Sho Diamond film biosensor
JP2000063154A (ja) * 1998-08-12 2000-02-29 Mitsubishi Chemicals Corp 核酸固定用ガラスプレート
EP1281967A2 (fr) * 1998-12-01 2003-02-05 Hitachi Software Engineering Co., Ltd. Biopuce et son procédé de production
EP1279960A1 (fr) * 2000-04-04 2003-01-29 Toyo Kohan Co., Ltd. Verre pour lame sur lequel une couche de traitement de surface est formee
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