JP2005077154A - Microarray, its manufacturing method, and microarray manufacturing device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To easily inspect the quality of a microarray. <P>SOLUTION: This microarray arranged with a plurality of spots on a substrate 100 includes the first spots 110 containing a biological substance, and second spots 112 arranged adjacent to the first spots and comprising a substance for inspection different from the biological substance. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a microarray in which a plurality of droplets of a biomolecule solution containing proteins, nucleic acids, and the like are arranged on a substrate, and a method for manufacturing the same.

  When analyzing proteins, nucleic acids, DNA, or the like, an analysis chip (microarray) in which a large number of biomolecule solutions containing biomolecules such as proteins are arranged on a substrate is used. Such a microarray is manufactured by spotting a biomolecule solution on a substrate using a contact pin, as disclosed in, for example, Japanese Patent Application Laid-Open No. 2000-287670 (Patent Document 1). In addition, there is a method of manufacturing a microarray by spotting a biomolecule solution on a substrate using a droplet discharge head (so-called inkjet head), which is disclosed in, for example, Japanese Patent Application Laid-Open No. 2002-253251 (Patent Document 2). Has been. The microarray described in Patent Document 2 discloses a technique in which two or more spots are formed with the same kind of biomolecule solution (probe solution), and one of them is used for product inspection.

JP 2000-287670 A Japanese Patent Laid-Open No. 2002-253251

  When manufacturing a wide variety of microarrays with stable quality, they can be manufactured as appropriate for substrates made of various materials, substrate surface conditions, and a large number of types of biomolecule solutions (sample solutions). It is necessary to control conditions and manufacturing environment, and to manage the quality.

  In the prior art described in Patent Document 2 described above, when a quality inspection is performed, a reagent is dropped only on a spot for product inspection, and a reaction is observed to detect a desired spot (probe) at a desired position. ) Was formed. For this reason, in order to drop the reagent only on the target spot and prevent the reagent from coming into contact with other spots, precise position control and control of the amount of reagent are necessary. However, considering mass production in particular, it is desirable that the quality inspection of the microarray be performed as easily as possible, and further technical improvements have been desired.

  Therefore, an object of the present invention is to provide a technique that enables simple quality inspection of a microarray.

  In order to solve the above-described problem, the microarray according to the first aspect of the present invention is a microarray in which a plurality of spots are arranged on a substrate, the first spot including a biological substance, and the first spot A second spot that is arranged adjacent to the spot and is made of a test substance different from the biological substance.

  The microarray according to the second aspect of the present invention is a microarray formed by arranging a plurality of spots on a substrate, and includes a first spot containing a biological substance and a dilution solvent, and adjacent to the first spot. And a second spot made of a dilution solvent, wherein the second spot is used for quality inspection.

  In these microarrays, a second spot that does not contain a biological substance and is configured to be suitable for quality inspection is appropriately disposed on the microarray adjacent to the first spot necessary for the original purpose of the product. By examining the presence or absence of a biologically related substance that is not originally contained in such a second spot, or the content thereof, it is possible to identify the presence or absence of contamination from the adjacent first spot. Since the degree of mutual contamination of the first spots formed on the substrate can be determined from the identification result, the quality inspection of the microarray can be easily performed. In particular, unlike the conventional method, precise position control and reagent amount control are performed, and the complicated operation of selectively dropping the reagent only on the inspection spot is not required, and quality inspection can be performed easily. It is suitable for mass production processes.

  Here, the term “microarray” in this specification refers to one in which droplets of many types of biomolecule solutions are arranged on a substrate made of glass or the like. The biomolecule solution is a solution containing DNA (deoxyribonucleic acid), protein, and the like, and is sometimes called a probe solution.

  In the microarray of each aspect described above, it is more preferable that a plurality of second spots are arranged around one first spot. As a result, since one first spot can be inspected using a plurality of second spots, the inspection accuracy can be improved.

  Moreover, it is preferable that the biological substance described above is fluorescently labeled. Thereby, the presence or absence of the biological substance in the second spot can be identified by an optical method, and the inspection is further facilitated. However, after spot formation, a reagent that is fluorescently labeled and binds to a biological substance contained in the first spot can also be used.

  The third aspect of the present invention is a method of manufacturing a microarray in which a plurality of spots are arranged on a substrate, wherein a first solution containing a biological substance and a dilution solvent is discharged onto the substrate. Discharging a second solution comprising a first spot forming step for forming one spot and a diluting solvent (or a test substance different from a biological substance) to a position adjacent to the first spot on the substrate; And a second spot forming step of forming a second spot at the position.

  With this manufacturing method, quality inspection can be easily performed, and a microarray suitable for a mass production process can be obtained.

  Moreover, it is preferable to further include an inspection step for inspecting whether or not the biological material is contained in the second spot. Thereby, the cross contamination between each spot is detectable.

  The above-described inspection process is more preferably performed by an optical method. That is, the biologically relevant substance contained in the first solvent described above is fluorescently labeled in advance or a fluorescently labeled reaction reagent is used, and in the inspection process, the light emission caused by the fluorescent label is detected to detect the second spot. It is preferable to perform an inspection. Thereby, an inspection process becomes still easier.

  The fourth aspect of the present invention is a microarray manufacturing apparatus for manufacturing a microarray in which a plurality of spots are arranged on a substrate, the first solution including a biological substance and a dilution solvent, and a dilution A discharge means that can individually discharge a second solution made of a solvent (or a test substance different from a biological substance), and a position control means that controls the relative position between the discharge means and the substrate. The means is configured by disposing the discharge hole for discharging the second solution adjacent to the discharge hole for discharging the first solution.

  With such a manufacturing apparatus, quality inspection can be easily performed, and a microarray suitable for a mass production process can be obtained. In particular, by arranging the discharge holes of the discharge means as described above, it becomes possible to efficiently manufacture the microarray according to the present invention, and not only on the substrate but also on the surface provided with the discharge holes. Cross contamination can be assessed.

  Hereinafter, embodiments of the present invention will be described in detail.

  FIG. 1 is a diagram illustrating a microarray according to an embodiment to which the present invention is applied. FIG. 2 is a plan view schematically showing the configuration of the microarray.

  As shown in FIG. 1, the microarray of this embodiment is configured by arranging a plurality of droplets (spots) containing a biological substance on a substrate 100 made of glass or the like. The spots formed on the substrate 100 include a first spot 110 containing a biological substance and a second spot 112 made of a test substance different from the biological substance. In the example of FIG. 1, the spots are arranged in a matrix, but other arrangements may be used.

  The first spot 110 is to be used for analysis or the like, which is the original purpose of a microarray product, and is arranged on the substrate 100 at a predetermined interval from each other. Examples of the biological material contained in the first spot 110 include proteins and nucleic acids. In addition, it is more preferable that the biological substance contained in the first spot 110 is fluorescently labeled using a fluorescent dye, and in this embodiment, this is the case.

  The second spot 112 is to be used for product inspection at the time of manufacturing the microarray, and the first spot 110 at the position is partially in the first spot 110 arranged in a matrix. It is arranged by replacing with. That is, the second spot 112 is disposed adjacent to the first spot 110.

  This second spot 112 only needs to contain no biological material. Therefore, any material can be used for forming the second spot 112 as long as it is a material different from the biological material and suitable for testing (test material). A suitable solvent for forming the second spot 112 is, for example, a dilution solvent such as a phosphate buffer (PBS). Regarding the second spot 112 formed in this way, it is possible to grasp the mutual contamination between the spots by examining the presence or absence of a biological substance in a later inspection process. In particular, when the biologically relevant molecule contained in the first spot 110 is fluorescently labeled, the presence or absence of the biologically relevant substance in the second spot 112 is detected by detecting luminescence resulting from the fluorescent label. It becomes possible and inspection becomes easy. Compared to a case where a second spot is not formed and a space is left, analysis by an image becomes easier, and since it is close to an actual use situation, an accurate microarray inspection can be performed.

  In the present embodiment, in FIG. 1, for example, as shown on the upper left side of the substrate 100, four second spots 112 are arranged around one first spot 110. Basically, the number of second spots is not limited as long as the second spots are arranged adjacent to the first spots 110, and may be one. Conversely, eight second spots 112 may be arranged around one first spot 110. When a plurality of the second spots 112 are arranged, it becomes possible to inspect the cross contamination in more detail. For example, it is possible to determine that cross-contamination is more likely to occur between spots adjacent in the horizontal direction than between spots adjacent in the vertical direction, and information effective in improving the manufacturing process can be obtained. Further, by inspecting with the plurality of second spots 112, the reliability of the inspection result is also improved.

  Next, a method for manufacturing the above-described microarray will be described.

  FIG. 2 is a diagram for explaining a manufacturing process of the microarray of this embodiment. A specific example of a manufacturing apparatus for manufacturing the microarray will be described later.

  On the substrate 100, for example, a first solution containing a biological substance such as protein and a dilution solvent is dropped to form a first spot (step S100). In addition, a second solution made of a diluting solvent is dropped on the substrate 100 to form a second spot (step S101). Here, about each process of step S100 and step S101, it is also possible to mutually replace these, and you may perform both in parallel (simultaneously). Each of the first and second spots is preferably formed by a droplet discharge method (so-called inkjet method). Thereby, each spot can be formed by controlling the discharge amount and the discharge position of the droplet with high accuracy.

  Next, using the second spot 112 formed on the substrate 100, an inspection for cross-contamination between the spots is performed (step S102). As described above, when the bio-related substance contained in the first spot 110 is fluorescently labeled and cross contamination occurs, the bio-related substance is contained in the second spot 112. In the present embodiment, the presence or absence of a biological substance is detected for each second spot 112 using a highly sensitive fluorescence detection device such as a laser scanner.

  In this inspection step, the total number of manufactured microarrays may be inspected, but some samples (for example, several) may be extracted from each manufacturing lot and inspected. By conducting such inspections, for example, if the number and extent of cross-contamination in a certain production lot is large, it can be determined that some defect has occurred during the manufacturing process and can be dealt with appropriately. It becomes.

  Next, a specific example of the microarray manufacturing apparatus will be described.

  FIG. 3 is a diagram (perspective view) for explaining the configuration of the microarray manufacturing apparatus of this embodiment. The microarray manufacturing apparatus shown in FIG. 3 is for manufacturing a microarray in which a plurality of spots are arranged on a substrate 100. The table 1, the Y-direction drive shaft 2, the droplet discharge unit 3, and the X-direction drive shaft 4 are used. , A drive unit 5, a control computer 6, and a CCD camera 7. The droplet discharge unit 3 corresponds to the discharge unit, and the table 1, the Y direction drive shaft 2, the X direction drive shaft 4, the drive unit 5, and the control computer 6 correspond to the position control unit.

  The table 1 is for mounting the substrate 100 constituting the microarray. The table 1 can mount a plurality of substrates 100, and is configured to be able to fix each substrate 100 by vacuum suction, for example.

  The Y direction drive shaft 2 is for freely moving the table 1 along the Y direction shown in the figure. The Y-direction drive shaft 2 is connected to a drive motor (not shown) included in the drive unit 5, and moves the table 1 by obtaining a drive force from the drive motor.

  The droplet discharge unit 3 discharges the biomolecule solution toward the substrate 100 based on the drive signal supplied from the control computer 6 so that the nozzle surface for discharging the solution faces the table 1. It is assembled to the X direction drive shaft 4. The droplet discharge unit 3 employs, for example, an inkjet head driven by an electrostatic drive method as a head for discharging a solution. The electrostatic drive type inkjet head has a relatively simple structure, stable solution discharge, and does not use heat, so it can avoid deterioration of biomolecules in the solution and maintain its activity. It becomes. In addition, the apparatus can be reduced in size and power consumption.

  FIG. 4 is a diagram illustrating a specific example of an ink jet head. In this figure, a perspective view of the ink jet head is shown, and the front side is partially a cross-sectional view showing an ink flow path and the like. The ink jet head of this embodiment includes a plurality of nozzles (ejection holes) 3a or 3b. Among these nozzles, the nozzle 3a is for discharging the first solution for forming the first spot, and is formed in two rows in the illustrated example. Each nozzle 3b is for discharging the second solution forming the second spot, and is arranged adjacent to the nozzle 3a. In the illustrated example, three nozzles 3b are arranged around one nozzle 3a. Thus, by arranging the nozzle 3b corresponding to the second solution adjacent to the nozzle 3a corresponding to the first solution, the first and second spots arranged adjacent to each other can be easily and It can be formed efficiently.

  Note that a piezo ink jet head using a piezoelectric element may be used as the droplet discharge head. In this case, similarly to the electrostatic driving method, there are advantages that heat is not used and that the discharge amount of the biomolecule solution can be easily adjusted by changing the applied voltage. Of course, as long as it is an inkjet system, any system such as a system that generates pressure in the flow path by heating, a system that uses an acoustic lens, or the like can be employed.

  The X direction drive shaft 4 is for freely moving the droplet discharge unit 3 along the X direction shown in the figure. This X-direction drive shaft 4 is connected to a drive motor (not shown) included in the drive unit 5 and moves the droplet discharge unit 3 by obtaining a drive force from the drive motor.

  The drive unit 5 includes a motor that drives the Y-direction drive shaft 2 and the X-direction drive shaft 4 and other drive mechanisms. By operating these motors and the like based on drive signals supplied from the control computer 6, the relative position between the table 1 on which the substrate 100 is placed and the droplet discharge unit 3 is controlled.

  The control computer 6 is installed in the housing of the drive unit 5 and controls the operation of the droplet discharge unit 3 (solution discharge timing, the number of discharges, etc.) and the operation of the drive unit 5. The control computer 6 also has a function of performing predetermined image processing on the image output from the CCD camera 7 and detecting the state of the droplet on the substrate 100.

  The CCD camera 7 is for capturing an image of a droplet of a biomolecule solution formed on the substrate 100 from above and obtaining an image. An image captured by the CCD camera 7 is output to the control computer 6.

  Thus, the microarray of this embodiment does not include a biological substance, and the second spot 112 configured to be suitable for quality inspection is adjacent to the first spot 110 necessary for the original purpose of the product. It arrange | positions suitably on a microarray. By examining the presence or absence of biologically related substances that are not originally contained in such second spots 112, the presence or absence of contamination from the adjacent first spots 110 can be identified. Since the degree of mutual contamination of the first spots 110 formed in large numbers on the substrate 100 can be determined from the identification result, the quality inspection of the microarray can be easily performed. In particular, unlike the conventional method, precise position control and reagent amount control are performed, and the complicated operation of selectively dropping the reagent only on the inspection spot is not required, and quality inspection can be performed easily. It is suitable for mass production processes.

  In addition, this invention is not limited to the content of the said embodiment, A various deformation | transformation implementation is possible within the range of the summary of this invention.

It is a figure explaining the microarray of one Embodiment. It is a figure explaining the manufacturing process of a microarray. It is a figure (perspective view) explaining the structure of a microarray manufacturing apparatus. It is a figure explaining the specific example of an inkjet head.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Table, 2 ... Y direction drive shaft, 3 ... Droplet discharge unit, 4 ... X direction drive shaft, 5 ... Drive part, 6 ... Control computer, 7 ... CCD camera, 100 ... Board | substrate, 110 ... 1st Spot, 112 ... second spot

Claims (8)

A microarray comprising a plurality of spots arranged on a substrate,
A first spot containing a biological substance;
A second spot that is arranged adjacent to the first spot and is made of a test substance different from the biological substance;
Including a microarray. A microarray comprising a plurality of spots arranged on a substrate,
A first spot comprising a biological material and a diluent solvent;
A second spot disposed adjacent to the first spot and comprising the diluent solvent;
And the second spot is used for quality inspection.   The microarray according to claim 1 or 2, wherein a plurality of the second spots are arranged around one first spot.   The microarray according to claim 1 or 2, wherein the biological substance is fluorescently labeled. A method for producing a microarray comprising a plurality of spots arranged on a substrate,
A first spot forming step of forming a first spot by discharging a first solution comprising a biological substance and a dilution solvent onto the substrate;
A second spot forming step of discharging a second solution comprising the diluting solvent to a position adjacent to the first spot on the substrate to form a second spot at the position;
A method for producing a microarray, comprising:   The microarray manufacturing method according to claim 5, further comprising an inspection step of inspecting whether or not the biological material is contained in the second spot.   The microarray manufacturing method according to claim 6, wherein in the inspection step, the second spot is inspected by detecting light emission caused by the fluorescent label. A microarray manufacturing apparatus for manufacturing a microarray having a plurality of spots arranged on a substrate,
A discharge means capable of individually discharging a first solution comprising a biological substance and a dilution solvent and a second solution comprising the dilution solvent;
Position control means for controlling the relative position between the discharge means and the substrate,
The discharge means is a microarray manufacturing apparatus, wherein a discharge hole for discharging the second solution is disposed adjacent to a discharge hole for discharging the first solution.