WO2004017069A1 - Analyseur a adn et systeme d'analyse en ligne - Google Patents

Analyseur a adn et systeme d'analyse en ligne Download PDF

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Publication number
WO2004017069A1
WO2004017069A1 PCT/JP2002/008312 JP0208312W WO2004017069A1 WO 2004017069 A1 WO2004017069 A1 WO 2004017069A1 JP 0208312 W JP0208312 W JP 0208312W WO 2004017069 A1 WO2004017069 A1 WO 2004017069A1
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WO
WIPO (PCT)
Prior art keywords
biochip
light
light source
exposure
illumination
Prior art date
Application number
PCT/JP2002/008312
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English (en)
Japanese (ja)
Inventor
Tohoru Hayashi
Toru Yamada
Shozo Ishizaka
Original Assignee
Kabushiki Kaisha Hayashi Soken
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
Application filed by Kabushiki Kaisha Hayashi Soken filed Critical Kabushiki Kaisha Hayashi Soken
Priority to JP2004528820A priority Critical patent/JP3691837B2/ja
Priority to PCT/JP2002/008312 priority patent/WO2004017069A1/fr
Publication of WO2004017069A1 publication Critical patent/WO2004017069A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

Definitions

  • a sample gene sample is labeled with a fluorescent dye, and this is reacted with a separate gene or gene product immobilized on a number of microspots on a biochip substrate, and the labeled sample is bound.
  • Production of biochips by the parallel access method which measures the position and degree of binding of biochips that have been analyzed ⁇ ⁇ ⁇
  • biochip analyzers that perform reactions and readings, in particular, genes that quantify genes or gene products in a sample It is used for analysis, as well as protein analysis and immunology-related analysis.
  • the present invention relates to a biochip information sensor having a database storing data necessary for biochip production and a program related to biochip analysis, and a biochip analysis device such as the Internet.
  • the present invention relates to a biochip online analysis system that can be connected via a network to efficiently perform analysis from biochip production. Background art
  • Biochip fabrication methods include photolithography, jet printing using piezoelectric technology, and mechanical microspot method. Have been.
  • the photolithography method is a technology for producing a biochip by using the exposure technology of a semiconductor device manufacturing method. Biochip fabrication by this technique selectively irradiates light through a photomask to the position of a small, predetermined sequence of oligonucleotides on a glass substrate, and the light that has passed through the unmasked area corresponds to four types of bases.
  • the phosphoramidite reagent is activated to cause a coupling reaction, and in each coupling step, a base is added one by one in a predetermined region to synthesize and extend an oligonucleotide directly on the substrate.
  • This method uses a photoactivation reaction to directly synthesize a large number of oligonucleotides with different sequences on microscopic spots on a substrate.Therefore, the biochip can be produced with the use of oligonucleotide sequence data. There is a place.
  • the above-described method has a disadvantage that it requires a large-scale apparatus similar to a semiconductor device manufacturing method, and therefore, at present, biochip users purchase and use a biochip manufactured by a specialized manufacturer. is there.
  • a miniaturized biochip manufacturing device is, for example, a digital micromirror device (hereinafter referred to as DMD).
  • DMD digital micromirror device
  • the one used is proposed in Japanese Patent Publication 2000-40660.
  • the DMD is a reflection type spatial light modulator using a micro mirror as an image element.
  • a 1280 x 1024 microphone opening mirror arrayed two-dimensionally is commercially available.
  • the micromirror uses an unstable state, ie, a state in which it does not tilt (OKF state of the power supply).
  • the method and apparatus proposed in Japanese Patent Application Laid-Open No. 2000-40660 include measures in an exposure apparatus used as a biochip producing apparatus for eliminating variations due to the position of the exposed surface and keeping the in-plane distribution of the exposed surface constant. Not done. Specifically, an expensive optical system using a fly-eye lens or the like must be installed outside, and another means must be provided to make the illumination light uniform.
  • biochip reading method a biochip reading apparatus
  • biochips manufactured by one photolithography method but also biochips manufactured by other methods are reacted with the fluorescently labeled test sample, and the fluorescence intensity of the sample bound to the micro spot on the biochip Is detected.
  • a confocal scanning fluorescence detector is used for this detection.
  • the confocal scanning fluorescence detection device has a small depth of focus and can exclude fluorescence emitted from a point other than an observation point out of focus, so that it is possible to read a biochip with high accuracy.
  • scanning must be performed by scanning the beam that excites fluorescence or by moving the sample itself.
  • the method of scanning the beam is that a flat detection field must be made using an F-theta lens, and that it is difficult to use an objective lens with a large numerical aperture.
  • the method of scanning the sample has a drawback that the mechanical and optical design is complicated because the flatness corresponding to the shallow depth of focus of the confocal system must be maintained.
  • an apparatus using a DMD as a spatial light modulator is disclosed as a scanning microscope in US Pat. No. 5,587,832 (JP-A-11-194275).
  • a main object of the present invention is to provide a biochip analyzer having the functions of biochip production, reaction, and reading by solving the problems of the conventional biochip analysis using the parallel assay method. .
  • Another object of the present invention is to provide a biochip analyzer that enables the production of a highly reproducible biochip using a spatial light modulator having an illumination light correction function for realizing uniform illumination. It is. Another object of the present invention is to provide a biochip analyzer that has high reliability and can perform high-speed scanning at high resolution by eliminating variations due to the positions of illumination excitation light and detection sensitivity when reading a biochip. It is to provide. Another object of the present invention is to collect data on the arrangement of spots of a plurality of nucleotides, peptides or sugar chains arranged on a biochip and the primary structure of nucleotides, peptides or sugar chains on the spots.
  • a biochip that distributes to a biochip analyzer through a combination network such as a network, and that collects and analyzes data read after reaction with a sample from the nanochip analyzer through a combination network and returns the results.
  • the biochip analyzer provides an exposure determined by the arrangement of a plurality of nucleotides, peptides, or sugar chains arranged on a biochip and the primary structure of the nucleotides, peptides, or sugar chains on the spots.
  • the pattern controls a single spatial light modulator illuminated by an exposure light source to selectively expose nucleotides, peptides, or sugar chains at positions on the biochip substrate, thereby allowing multiple types of bases or
  • a fluorescent excitation light source that forms an image on the spatial light modulator and selectively illuminates a small area on the biochip that has reacted with the test sample.
  • the biochip online analysis system includes a biochip analyzer as described above, a server device that performs analysis based on a data base that stores the sequence data of the biochips and the analysis data and the biochip read data. These devices are equipped with a biochip information sensor, and these devices are connected via a computer network.
  • the nanochip analyzer uses the biochip array data owned by itself or the biochip array data provided by the server device on demand to produce, react, and read biochips. Collects the biochip reading data from the biochip analyzer, analyzes it, and returns the analysis data to the biochip analyzer.
  • FIG. 1 is a system configuration diagram of an optical system and a control system of a biochip analyzer using a DMD according to a first embodiment of the present invention.
  • FIG. 2 is a system configuration diagram of an optical system and a control system of a biochip analyzer using a reflective liquid crystal panel according to a second embodiment of the present invention.
  • FIG. 3 is a system configuration diagram of an optical system of a biochip analyzer capable of analyzing at two wavelengths according to a third embodiment of the present invention.
  • FIG. 4 is a diagram showing a focal position when a DMD is illuminated with converged light.
  • FIG. 5 is a diagram showing a focal position when a DMD is illuminated with parallel light.
  • Fig. 6 is a conceptual diagram of the online analysis system and a flow chart from the production to the analysis of the biochip. BEST MODE FOR CARRYING OUT THE INVENTION
  • the biochip analyzer in the first embodiment can perform a series of steps of biochip preparation, reaction with a test sample, and reading.
  • the exposure for biochip fabrication and the illumination for fluorescence excitation for biochip reading are configured with a common optical system, and the exposure light monitor for biochip fabrication and the imaging and excitation light monitoring for biochip reading are shared. It is a biochip analyzer composed of the above optical system.
  • a biochip 1 At the center of the optical system, a biochip 1, an objective lens 2, an imaging lens 3, and a spatial light modulator DMD 6 are provided, and a central optical axis is formed.
  • the objective lens 2 and the imaging lens 3 operate as follows when producing and reading the biochip 1.
  • the biochip 1 is illuminated and exposed during production, the surface of the DMD 6 is imaged on the biochip 1 during reading, and the fluorescence emitted from the spot of the biochip 1 is imaged on the surface of the DMD 6.
  • the light from the light source 20 is incident on the fiber optic plate 24 whose exit end is cut obliquely, and a planar light source is formed at the exit end.
  • the real image of the planar light source forms an image on the mirror surface of the DMD 6 via the dichroic mirror 26 by the lens function of the lens 25.
  • the projection end of the fiber optic plate 24, the lens 25 and the mirror surface of the DMD 6 satisfy the requirements of Scheimpflug.
  • an imaging system in which a lens is added with an orientation so as to satisfy the condition of Scheimpflug is employed.
  • an imaging system in which the lens is shifted may be employed.
  • the light source 20 is composed of a light source A20, a light source B20, and a light source C20, and is capable of exciting two wavelengths of fluorescence at the time of reading and exposing at the time of fabrication.
  • Mirror 23 (23A, 23B) is installed to switch the three light sources. When the mirror 23A is introduced at a predetermined position, the light source A20 is used, and when the mirror 23B is introduced at a predetermined position (solid line in the figure), the light source B20 is used. Further, the mirrors 23A and 23B are retracted to the positions indicated by the broken lines in the figure, and when neither mirror is introduced, the light source C20 for exposure is used.
  • the light source of each wavelength of the above optical system can be added as needed.
  • the planar light source can also be constituted by a surface illuminant such as electroluminescence (EL).
  • EL electroluminescence
  • Z direction the image in the sample thickness direction
  • a thin-film scatterer with an extremely thin scattering layer, illuminated with predetermined light can also be used as a planar light source.
  • the thin film scatterer for example, Light Shaping Duffuser (POC, CA, Torrance, USA) is known.
  • the planar light source forms an image on the spatial light modulator and illuminates it, and images are taken corresponding to these pixels, one or more light emitting points are provided for each pixel of the spatial light modulator. If this is the case, it is possible to use an on-plane array of discrete point light sources, even if it is not a continuous surface light emitter such as an EL element.
  • An on-surface array is, for example, There are two-dimensional arrays of LEDs and Laser Diodes, and two-dimensional arrays of microphone aperture lenses and pinholes illuminated in a predetermined manner.
  • the on-plane array can be regarded as a fine and sufficient point light source, and can be configured with a two-dimensional array of point light sources that correspond one-to-one or one-to-one to the pixels of the spatial light modulator by combining with a reduction optical system I do.
  • predetermined light is incident from one end of an optical fiber bundle in which a number of optical fibers are bundled, and the light emitting end on the opposite side, similarly, the light emitting end of a fiber optic plate, and similarly the plexite
  • the light exit end can be used as a planar light source.
  • 16 to 25 point light sources which are emission ends of one fiber, are allocated to one pixel of the spatial light modulator.
  • One pixel of the spatial light modulator is a 16 / square micromirror in the case of DMD. In the case of a reflective liquid crystal panel, it is a 13.5 square liquid crystal element.
  • the imaging system includes an imaging lens 27, a filter 28 (28A, 28B, 28C), and a CCD 29, and captures a real fluorescence image of the biochip 1.
  • the filter 28 is selectively installed between the imaging lens 27 and the CCD 29, and transmits a wavelength corresponding to a fluorescent dye.
  • a fluorescent dye For example, FITC, Cy3, and Cy5 fluorescent dyes have bandpass filters with center wavelengths of 520 nm, 570 nm, and 670 nm, respectively.
  • DMD6, imaging lens 27 and CCD 29 satisfy the Scheimpflug requirements. This imaging system is also used for making the excitation light uniform or the exposure light uniform.
  • Computer System 3 is a software for controlling the DMD 6 necessary for the analysis of the biochip 1 for parallel assembly, chemical solution management, temperature management, CCD 29 control, and illumination light correction to equalize exposure light and excitation light.
  • the thermochemical solution supply system 4 includes a flow cell 41 in which the mounted biochip 1 forms an upper lid and maintains a set temperature, a reaction reagent container 42 in which an electromagnetic valve 44 controlled by the computer 30 is installed, and a cleaning solution container. 43. It is composed of a means for introducing a test sample, and supplies a chemical solution and a cleaning solution in synchronization with each step of the photoactivation force coupling reaction during biochip fabrication.
  • the flow cell 41 is used not only at the time of producing the biochip 1 but also at the time of reaction with a test sample and reading. When the above reaction is used for gene analysis, it is particularly called hybridization.
  • the test sample is introduced into the flow cell 41 from the inlet 45 and reacted.
  • the reading is performed with the biochip 1 in a wet state or a dry state.
  • the reading in the wet state is characterized by high fluorescence yield, and is performed by exposing the biochip 1 to a liquid.
  • the reading in the dry state is performed by introducing clean dry nitrogen gas from the inlet 45 and drying the biochip 1.
  • the temperature of the flow cell 41 can be set to a predetermined value, and the influence of a temperature change when reading the biochip 1 is examined. Also, it is necessary to use the entire device at an angle of 10 to 20 degrees. Adjustment screws (not shown) are provided. This is to expel the bubbles generated in the flow cell and to analyze Biochip 1 stably.
  • the analysis operation of the biochip 1 can be carried out entirely without attaching and detaching the biochip 1 from the flow cell 41, so that the efficiency can be improved and the contamination is reduced.
  • the DMD6 adopted as the spatial light modulator is a minute light deflecting element, and has a structure in which a number of minute microphone aperture mirrors are arranged.
  • Each micromirror can be individually tilted by two predetermined angles by digital control means. That is, each micromirror has two stable states, and has the function of switching light introduced from the outside to the optical path in two directions.
  • DMD6 has micromirror rooster 3 systems!], Such as 800X600, 1024X768, 1280X1024, 1920X1080, and two types of tilting angles of micromirrors: ⁇ 10 degrees and soil 12 degrees. , Either can be applied.
  • DMD6 exposure by pattern (at the time of fabrication) and confocal scanning fluorescence detection (at the time of reading) have been realized.
  • DMD6 is used for both biochip fabrication and biochip reading.
  • the light source unit 20 is switched to the exposure light source C.
  • the light from the exposure light source C enters the fiber optic plate 24, and the planar light source formed at the exit end forms and illuminates the DMD 6 for image formation.
  • This imaging illumination light is reflected by the DMD 6 micromirror tilted corresponding to the exposure pattern.
  • the reflected light forms an image via the imaging lens 3 and the objective lens 2, and a predetermined spot position of the biochip 1 is exposed.
  • a minute area of the second light source is cut out by the tilted micromirror, and a reduced image is formed on the biochip by the optical path formed by the imaging lens 3 and the objective lens 2.
  • the fluorescent light emitted from this imaging point reverses the above optical path and forms an image on a micro mirror.
  • the image on the micromirror is selected only for fluorescence by the dichroic mirror 26, formed again by the imaging lens 27, and captured by the CCD 29. At this time, the excitation light mixed in the fluorescent filters 28 (28A, 28B, 28C) is excluded to increase the S / N.
  • the reduction optical system including the objective lens 2 and the imaging lens 3 is employed, but the present invention is not limited to this.
  • the optical system an equal-magnification optical system or a magnifying optical system is selected according to the size of the biochip 1.
  • a uniform fluorescent plate is placed on the flow cell 41, and the fluorescence intensity corresponding to each micromirror is monitored by the CCD 15 to measure the intensity distribution of the exposure light before uniformization.
  • an exposure light correction table is created by calculating the reciprocal of the fluorescence intensity for each micromirror so that the exposure light intensity at the position corresponding to each micromirror becomes a constant value.
  • the following two methods use an exposure light correction table.
  • the first method controls the exposure time for each micromirror.
  • the exposure light correction table is referenced, and the exposure of each micromirror is controlled. Light time is variably controlled to shorten or extend.
  • the exposure time is kept constant in all microphone aperture mirrors, but intensity modulation is performed by applying binary pulse width modulation (BPWM) to each microphone aperture mirror.
  • BPWM binary pulse width modulation
  • the exposure light correction table is referred to, the intensity of the reflected light from each micromirror is corrected, and uniform exposure light is obtained as a whole.
  • This exposure light correction method is used not only when manufacturing a biochip, but also when reading as described below.
  • the number of micromirrors to be scanned is described as one, but the present invention is not limited to this.
  • the micromirrors are far enough apart on the DMD 6 to maintain the spatial light modulation function that removes defocused light and stray light, multiple microphone aperture mirrors can be translated and scanned simultaneously. can do. For example, high speed can be achieved by translating and scanning 2000 micromirrors.
  • the confocal fluorescence scanning means and the excitation light correcting means are simultaneously realized by a single spatial light modulator.
  • the intensity distribution of the illumination light at the time of reading is adjusted by a (uniform) uniform fluorescent screen.
  • the fluorescence intensity before homogenization is measured and an excitation light correction table is created.
  • Excitation light correction tables are provided separately for the number of illumination light sources. These excitation light correction tables are referred to according to the light source used at the time of reading, and the first method or the second method of exposure light correction corrects the variation of the illumination light and the detection sensitivity depending on the position.
  • the above correction function is realized by a computer and the software mounted on it.
  • the second embodiment is a biochip analyzer using a reflective liquid crystal panel called LCOS (Liquid Crystal on Silicon) as a spatial light modulator.
  • LCOS Liquid Crystal on Silicon
  • each pixel is a minute liquid crystal cell, and the polarization direction of light reflected by an applied voltage (analog amount) is controlled.
  • PBS Polarized Beam Splitter
  • the reflection type liquid crystal panel for example, integrates 1365 ⁇ 1024 liquid crystal cells having a size of 13.5 ⁇ 13.5 ⁇ .
  • a transmissive liquid crystal can be used. In the transmissive liquid crystal panel, the effective pixel area cannot cover the entire surface because some of the pixels are occupied by switching elements such as TFTs. Therefore, it is not possible to control the whole screen smoothly. It is necessary to design in consideration of this.
  • FIG. 2 shows the optical system of the second embodiment, in which the computer system and the constant temperature chemical liquid supply system shown in FIG. 1 are omitted.
  • a light source 20C for exposure used when manufacturing biochips and a light source 20A for excitation used when reading biochips are installed. These light sources are configured to be switched by introducing a mirror 23C into the optical path. That is, the mirror 23C is removed from the optical path and the exposure light source 20C is used during the fabrication of the biochip, and the mirror 23C is introduced into the optical path and the excitation light source 20A is used when reading the biochip.
  • Light from any of the foregoing light sources is introduced into the fiber optic plate 24, producing a planar light source at its exit end.
  • An image forming lens 25 for forming an image of the planar light source on the reflective liquid crystal panel 60, a dichroic mirror 26 for transmitting the fluorescent light and reflecting the excitation light and the exposure light, and a PBS 61 for splitting the light according to the polarization angle are provided. ing.
  • An imaging lens 3 and an objective lens 2 are provided between biochip 1 and PBS61.
  • the surface of the reflective liquid crystal panel 60 is imaged on the surface of the biochip 1 via the PBS 61.
  • the CCD 29 is used for capturing a fluorescent image or monitoring illumination light.
  • the image of the surface of the reflective liquid crystal panel 60 is formed on the CCD 29 by the imaging lens 27.
  • the polarization angle of the reflected light from the liquid crystal cell is controlled by the analog voltage applied to the liquid crystal cell.
  • the amount of light is controlled by the branching action of the PBS61 light transmitted or reflected by the controlled light polarization angle (specifically, when no voltage is applied to the liquid crystal cell, the light from the light source passes through the PBS61, Illuminate the biochip Apply a predetermined maximum voltage to the liquid crystal cell In this case, the light from the light source is reflected by PBS61 and does not illuminate the biochip.
  • the biochip is illuminated with a light amount that is inversely proportional to the applied voltage.
  • the spatial light modulation function is realized by controlling the voltage of the individual liquid crystal cells described above.
  • the spatial light modulation function using a liquid crystal cell is a mask function that generates an exposure pattern when manufacturing a biochip or a confocal fluorescence scanning function when reading a biochip, or an illumination light correction that is used for both when manufacturing and reading a biochip. Used as a function.
  • Each of these functions is the same as in the first embodiment, and a description thereof will be omitted.
  • the third embodiment is a biochip analyzer capable of performing two-wavelength analysis without performing mechanical switching.
  • RNA from a lesion in the same organ and RNA from a normal site are extracted, and cDNA obtained by reverse transcription of RNA at each site is labeled with two types of fluorescent dyes with different wavelengths, such as Cy3 and Cy5, and then mixed. Then, they are made to react with a single biochip, and the ratio of each is compared.
  • FIG. 3 shows the optical system of the third embodiment, in which the computer system and the thermostatic liquid supply system are omitted.
  • the present embodiment is a two-wavelength readable biochip analyzer in which two sets of the optical system for illumination for fluorescence excitation and the optical system for imaging of the first embodiment are arranged independently for each wavelength.
  • a biochip 1 In the center of the optical system, a biochip 1, an objective lens 2, an imaging lens 3, and a DMD 6 which is a spatial light modulator are installed, and a central optical axis is formed. versus The object lens 2 and the imaging lens 3 illuminate and expose the biochip 1 when producing the biochip. Further, at the time of reading, the surface of the DMD 6 is imaged on the biochip 1 to excite a predetermined spot, and the fluorescence emitted from the excited spot is imaged on the DMD 6 surface.
  • the optical system A constitutes an illumination system for exposing light with a light source C in addition to an illumination system for fluorescence excitation of a specific wavelength by a light source A and an imaging system.
  • the B optical system forms an illumination system and an imaging system for fluorescence excitation of a different wavelength from the A optical system.
  • Both optical systems consist of a light source A for fluorescence excitation: B, a light source C for exposure illumination, dichroic light sources 26A and 26B, and a non-pass-filled light source 28A and 28B with different optical elements for each wavelength. Have been. Other elements are the same. Here, only the A optical system will be described.
  • the optical system A is provided with a switchable light source A of a specific wavelength and a light source C of exposure light.
  • the light from the switched light source is made incident on the fiber optical plate 24, and a planar light source generated at the exit end thereof. Is formed on the surface of the DMD 6 by the illumination lens 25.
  • a dichroic mirror 26A for reflecting excitation light or exposure light and transmitting fluorescence is provided between the illumination lens 25 and the DMD 6.
  • the light source C is switched to produce a biochip in the same manner as in the first embodiment.
  • the biochip 1 is read by the switched excitation light having the specific wavelength of the light source A in the same manner as in the first embodiment.
  • the biochip 1 is subjected to confocal fluorescence scanning, and the fluorescence information of the biochip 1 can be read. The reading of a biochip using both the A optical system and the B optical system will be described based on a specific example.
  • the optical system A performs reading using Cy3 as a fluorescent dye and the optical system B performs reading using Cy5
  • the light sources A and B for fluorescence excitation and the dichroic mirrors 26A and 26B Bandpass Fillers 28A and 28B with wavelengths of 570 nm and 670 nm, respectively, are installed.
  • Bandpass Fillers 28A and 28B with wavelengths of 570 nm and 670 nm, respectively, are installed.
  • set up a uniform Cy3 fluorescent plate in place of the biochip and create a correction table for the A-optical system.
  • Biochip 1 is installed. First, the fluorescent information of Cy3 is read by the optical system A, and then the fluorescent information of Cy5 is read by switching to the optical system B.
  • a client for producing, reacting, and reading a biochip using the biochip analyzer of each of the above-described embodiments, and providing the client with the data necessary for the production of a biochip and providing the client with the data
  • the present invention provides a biochip analysis online system in which a biochip information sensor for performing a biochip analysis required by a client based on a reading data is connected to a computer network.
  • This system provides, through a computer network, a set of oligonucleotide sequence data--a set of protein amino acid sequence data and the primary structure of nucleotides, peptides or sugar chains. , Collect the read results from the client and return the analysis results to the client again.
  • the on-line system charges for providing information necessary for biochip fabrication and for analyzing based on the data read.
  • the biochip analyzer of the present invention is installed on the client side.
  • the biochip information center has a system with a built-in analysis program, a database for storing various biochip production data and a biochip analysis data.
  • the system itself has a built-in processing program for biochip production and a processing program for biochip analysis, a biochip production database, and an analysis database.
  • the biochip preparation data pace includes the base sequence set for amino acid sequence set for gene related analysis and protein analysis related to biochip preparation, and the sequence layout of spots on the biochip. Is stored in a predetermined data structure.
  • the biochip analysis database contains information on the classification of known genes and proteins that are referred to when performing analysis.
  • the client connects the biochip analyzer to the biochip information sensor, communicates with the biochip production processing program through the network, Request a list Listed biochips, for example, the Select and order biochips according to the analysis items, such as biochips.
  • the nanochip information center calculates the charge based on the biochip received from the client and the analysis items. After that, biochip production data consisting of the base sequence data set of each spot of the selected biochip and the sequence layout of the spots on the biochip is transmitted, and the fee is notified.
  • the biochip analyzer receives the biochip production data, it performs biochip production.
  • the test sample is introduced into the prepared biochip from the test sample inlet manually or automatically upon receipt of a trigger signal for completion of the biochip, and a reaction such as hybridization is performed. Receiving a trigger signal to end the reaction of the biochip, it automatically starts reading the biochip and stores the read data in memory.
  • the client requests the Biochip Information Center for analysis by designating a biochip.
  • the biochip information center activates the biochip analysis processing program in response to the client's request, and transmits the analysis item list to the client.
  • the analysis item list provides various processing and analysis items such as visualization processing, class analysis, structural hierarchy analysis, and polymorphism analysis.
  • the client selects a predetermined item from the displayed analysis item list, places an order, and transmits the biochip read data.
  • the biochip information sensor receives an order for a specified biochip analysis item, calculates billing, receives biochip read data, performs processing and analysis based on the selected analysis item, and transmits the result.
  • the client receives the analysis result and the billing information, displays the confirmation information, confirms the content, and stores it in the memory.
  • This system does not limit the biochip analyzer of the present invention alone to a client.
  • the present invention can also be used with the biochip analyzer and biochip reader of the present invention, or other existing biochip fabrication or read-only machines compatible with networks.
  • this system connects the biochip analyzer of the present invention to a network and uses it in various forms such as analysis by reading biochips prepared by other devices or analysis from biochip read data prepared in advance. can do.
  • a recording medium such as a CD or a DVD
  • the client can communicate with the biochip analysis-compatible processing program immediately after communicating with the biochip manufacturing-compatible processing program to perform the order of production and analysis.
  • a series of analysis using a biochip for reading and analysis is performed automatically. This fully automatic operation is to be performed without the intervention of an operation in the middle of the above series of analysis operations.
  • the present invention is not limited to the biochip analyzer of the above-described embodiment, but can be applied to an apparatus constituted by any or a combination of the functions of biochip production, reaction, and reading.
  • the present invention uses the conventional southern plot and northern plot methods in addition to biochips, instead of radioisotope labeling and detection, with fluorescent dyes. It can be applied to the measurement of fluorescence intensity distribution when performing labeling and detection.
  • the above-mentioned southern blot and northern blot method is an analysis method using hybridization.
  • the above-mentioned applications include, for example, measurement of the fluorescence intensity distribution of nylon, ditrocellulose membrane / gel membrane, and the like.
  • it can be used not only for measuring the fluorescence intensity distribution but also for measuring the intensity distribution of chemiluminescence in experiments in which labeling was performed with a chemiluminescent substance that does not require excitation light.
  • a sample that emits chemically and uniformly is used instead of the uniform fluorescent plate used in the present embodiment.
  • the spatial light modulator can be realized by software control without using a photomask used in the conventional photolithography method.
  • exposure light correction unlike the conventional method of equalizing illumination by hardware using a fly-eye lens or the like, the present invention corrects exposure light for each individual device, thereby eliminating variations between devices. At the same time, it is possible to flexibly cope with switching and replacement of the light source.
  • the exposure light correction means can be added to the spatial light modulation element that generates the exposure pattern, thereby contributing to downsizing and cost reduction of the apparatus.
  • the reading of the biochip can be realized by controlling the conventional confocal scanning function using software using a spatial light modulator. Also, the in-plane distribution of the illumination light is improved by adjusting the intensity of the illumination light for each spatial light modulator, and the variation in the illumination light and the detection sensitivity depending on the position is corrected.
  • the biochip analyzer on the client side and the system main body of the biochip information sensor are connected via a computer network.
  • the client receives the sequence information of the biochip ordered from the biochip information center online and the set information of the primary structure data by the biochip analyzer.
  • the biochip analyzer performs the steps of biochip preparation, reaction, and reading based on the above set information, and the biochip read data is collected by biochip information sensor and specified by the client. Perform biochip analysis according to the analysis items.
  • the analysis data is sent to the client. This allows the client to analyze the test sample on hand.

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Optics & Photonics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

Selon l'invention, un photoconvertisseur d'espace unique (6) éclairé à l'aide d'une source de lumière d'exposition C (20) est commandé en fonction du modèle d'exposition qui est déterminé selon la topologie de plusieurs points de nucléotides, peptides ou chaînes du sucre situés sur une puce à ADN (1) et les structures primaires des nucléotides, peptides ou chaînes du sucre sur les points, de telle manière que les emplacements des points des nucléotides, peptides ou chaînes du sucre sur le substrat de la puce à ADN sont sélectivement exposés à la lumière. Ainsi, des réactions de couplage photoactivées sont induites à des endroits définis de plusieurs types de bases, d'acides aminés ou de saccharides et les nucléotides, peptides ou chaînes du sucre sont synthétisés et étendus pour produire une puce à ADN. Puis, le photoconvertisseur d'espace est éclairé au moyen des sources de lumière d'excitation fluorescentes A et B (20), afin de lire les données de la puce à ADN. L'utilisation du dispositif susmentionné par un client repose sur des centres de traitement de l'information de puce à ADN connectés à un réseau d'ordinateurs. Ainsi, l'analyse et la construction de la puce à ADN en ligne peuvent être réalisées.
PCT/JP2002/008312 2002-08-16 2002-08-16 Analyseur a adn et systeme d'analyse en ligne WO2004017069A1 (fr)

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JP2004528820A JP3691837B2 (ja) 2002-08-16 2002-08-16 バイオチップ分析装置
PCT/JP2002/008312 WO2004017069A1 (fr) 2002-08-16 2002-08-16 Analyseur a adn et systeme d'analyse en ligne

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JP2014092682A (ja) * 2012-11-02 2014-05-19 Olympus Corp 顕微鏡用照明装置及びそれを備えた顕微鏡
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