JP2002267666A - Method and apparatus for manufacturing test chip - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method and an apparatus for manufacturing a test chip, wherein cDNA spots to be arranged on the test chip are arranged with satisfactory accuracy in a matrix shape. SOLUTION: A cDNA-containing solution 32 as a droplet is sent out into a carrier solution 22 sent into a nozzle 12. Fig. (a) shows the coating start state of the carrier solution 22, the nozzle 12 is lowered, and the droplet of the carrier solution 22 is increased gradually at the lower end. Fig. (b) shows a state that the nozzle 12 is stopped in a contact position, the carrier solution 22 is increased, and it comes into contact with the surface of a substrate 20 so as to be set to a state in Fig. (c). Fig. (d) shows a state that the nozzle 12 is raised so as to cut the solution and that the nozzle is moved to the plane direction of the substrate 20. The carrier solution 22 on the substrate 20 is volatilized, spots 50 of the cDNA-containing solution are formed, and a next coating start state is set as shown in Fig. (e). When the operations in Figs. (a) to (e) are repeated, the test chip in which the spots 50 are arranged in the matrix shape on the substrate 20 is obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and an apparatus for producing a test chip used for biological analysis.

[0002]

2. Description of the Related Art In recent years, the technology in the field of genetic engineering has been rapidly developed, and the human genome project has been developed with one object of decoding the base sequence of the human genome, which is thought to reach 100,000. . Biological analysis is used for these projects. Biological analysis includes, for example, immunoassay using an antigen or antibody utilizing high specificity of antigen-antibody reaction, gene analysis using so-called hybridization, genetic diagnosis, biotin-avidin method, etc. There is an analysis that uses a dynamic combination.

On the other hand, enzyme immunoassays and fluorescent antibody methods utilizing antigen-antibody reactions have been used for diagnosis and research, and research has been advanced to search for DNAs affecting various genetic diseases. Microarray technology has attracted attention as one of the methods.

[0004] The microarray technology uses a microarray chip (a test chip and a test chip) in which a large number of different known cDNAs which have already been decoded are coated in advance in a matrix form on a membrane filter or a slide glass at a high density (interval of several 100 μm or less). And sometimes referred to as a DNA chip). cDNA is a DNA complementary to messenger DNA (mDNA).
Refers to A chain. Further, a test chip is known which uses a biological substance, for example, an antibody, an antigen, a substrate, DNA, mDNA or the like as a specimen.

For example, a healthy person A labeled with a fluorescent dye a
DNA taken from a cell (an example of an organism-derived substance)
DNA obtained from the cells of the specimen B having the genetic disease labeled with the fluorescent dye b, the DNA of each specimen dropped on this microarray chip with a pipette or the like, and after the cDNA on the microarray chip is hybridized, Each cDNA on the microarray chip is scanned with excitation light for separately exciting each of the fluorescent dyes a and b, and each fluorescence of each cDNA is detected by a photodetector. Based on the detection result associated with the fluorescence emission position on the microarray chip, the DNA of each sample is determined as to which cDNA has been hybridized, and by comparing the cDNA hybridized between both samples, DNA expressed or deficient in the above disease is specified. Here, the terms “hybridization” and “hybridization” mean the formation of a double strand of a complementary base sequence between DNA fragments, and also broadly include specific binding.

[0006] By the way, the microarray chip has a very large number of cDNAs coated on a substrate such as a slide glass as described above.
One needle-shaped coating tip is immersed in a large number of cDNAs prepared in advance, and the cDNA attached to the tip of the coating tip is applied to a predetermined position on a slide glass in a dotted manner, and then the coating tip is washed and dried. Then, the so-called spotting is performed by repeating the operation of dipping in the next different kind of cDNA and applying it to another predetermined position on the slide glass.

[0007]

However, a cDNA which is a specific binding substance to be coated on a slide glass or the like
Can be tens of thousands, and it takes a lot of time to manufacture one macro array chip. For this reason, the price of the macrochip is extremely high, which hinders the promotion of the above-described DNA expression research and the application to screening using immunological analysis.

The present invention has been made in view of the above circumstances, and has as its object to provide a method and an apparatus for producing a test chip used for biological analysis such as DNA analysis and immunological analysis.

[0009]

According to a method of manufacturing a test chip of the present invention, a liquid containing a specific binding substance is intermittently sent as droplets to a flowing carrier liquid, and the carrier liquid is supplied from a nozzle to a substrate. When arranged on the substrate, the carrier liquid containing the specific binding substance is arranged on the substrate.
After the carrier liquid containing a specific binding substance droplet is brought into contact with the substrate by reducing the distance between the nozzle tip and the substrate, the carrier containing the specific binding substance droplet is increased by increasing the distance between the nozzle tip and the substrate. Preferably, the liquid is transferred from the nozzle tip to the substrate. Further, when arranging the carrier liquid containing the specific binding substance on the substrate, it is more preferable to apply an electrostatic field to the carrier liquid containing the droplet of the specific binding substance.

[0010] The method of manufacturing a test chip according to the present invention comprises:
The liquid containing the specific binding substance is intermittently sent as droplets to the flowing carrier liquid, the carrier liquid is sent out from the nozzle onto the substrate, and the carrier liquid containing the specific binding substance is put on the substrate. When arranging, it is preferable to apply an electrostatic field to the carrier liquid containing the droplet of the specific binding substance.

Preferably, the carrier liquid is an organic solvent, and the solvent of the liquid containing the specific binding substance is water. Further, the organic solvent is an alkane, a ketone,
It is preferable to be composed of at least one of esters. Further, the specific binding substance is cDN
It is preferably one of the substances having the function of A. It is preferable that the substance having the function of cDNA is a substance having the function of a plurality of different types of cDNA.

More preferably, the interfacial tension between the carrier liquid and the liquid containing the specific binding substance is 1 mN / m or less.

It is preferable to add a surfactant to the solution containing the specific binding substance. More preferably, the surfactant contains at least one of sulfosuccinates and gluconiamides. Further, it is preferable to add a salt to the solution containing the specific binding substance.
The salt is a salt formed by combining an alkali metal ion as a cation and a polyvalent carboxylate ion, a halide ion or an inorganic acid ion as an anion, and more preferably contains at least one of them.

According to the test chip manufacturing apparatus of the present invention, in a test chip manufacturing apparatus in which a specific binding substance is arranged on a substrate, a pipe through which a carrier liquid flows, means for feeding the carrier liquid into the pipe, Droplet sending means connected to a tube and intermittently sending a liquid containing the specific binding substance as a droplet into the carrier liquid, a nozzle provided at the tip of the tube, and a holding member for holding the substrate A first position in which the substrate of the holding member or the nozzle is moved toward and away from each other to bring the nozzle tip close to the substrate so that the carrier liquid at the nozzle tip contacts the substrate; and Liquid draining means for alternately displacing the carrier liquid containing the droplet of the specific binding substance away from the substrate to a second position where the carrier liquid is transferred from the nozzle tip to the substrate as a droplet,
An arrangement means for relatively moving the nozzle or the holding member in the plane direction of the substrate in conjunction with the liquid draining means and arranging the droplets of the specific binding substance on the substrate. Further, it is more preferable to have high voltage applying means for applying a DC high voltage to the holding member. Further, the liquid containing the specific binding substance is a plurality of liquids,
More preferably, the droplet sending means switches the plurality of kinds of liquids and sequentially sends them out.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION [Carrier Liquid] The carrier liquid is c
It is intended to transport droplets containing specific binding substances such as DNA, and the flow is preferably a plug flow. It does not mix with the solution or dispersion of the specific binding substance, or it is dried before mixing What removes, ie, what can maintain a phase separation state for at least 1 minute or more is preferable. Further, the carrier liquid may be composed of only a liquid that can be removed, preferably evaporated or volatilized under conditions that do not decompose or alter the specific binding substance. A solution in which a solute is dissolved using this liquid as a solvent may be used.

If the carrier liquid and the solvent of the specific binding substance are the same (eg, water), diffusion occurs, but if the flow is a plug flow, the mass transfer rate of the diffusion is lower than the mixing rate, so that substantially no mixing occurs. Absent. However, since the specific binding substance is usually injected into the carrier liquid as an aqueous solution, the carrier liquid is preferably an organic solvent having low solubility in water. Examples of preferred organic solvents include ketones, esters, alkanes and the like. Specifically, alkanes (eg, n-pentane, n-heptane, cyclohexane, etc.), ketones (eg, methyl ethyl ketone), esters (eg, methyl formate, ethyl formate, methyl acetate,
Ethyl acetate).

In order to make the specific binding substance a microdroplet in the carrier liquid, the interfacial tension between the specific binding substance and the carrier liquid is preferably 1 mN / m or less, more preferably 0.5 mN / m. m or less.

[Specific binding substances] Specific binding substances are hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNAs, DNAs, and Rs.
NA, mRNA, etc., means a substance that can specifically bind to a biological substance. This specific binding substance is preferably in a form held on a carrier. Known carriers such as gelatin particles, polymer latex such as polystyrene latex, and inorganic particles such as silica gel can be used as the carrier. Retention of the specific binding substance and the carrier is carried out by covalent
It is performed by ionic bonding, physical adsorption, or the like.

The specific binding substance is usually dissolved in water and used as an aqueous solution of the specific binding substance for producing a test chip, but the solvent for dissolving the specific binding substance is not limited to water. As long as the specific binding substance is not deteriorated, a known solvent can be used. Surfactants can be added to reduce the interfacial tension of the specific binding agent solution. Further, when the specific binding substance reacts with the substrate and is immobilized on the surface of the substrate, a salt exhibiting a pH buffer function may be added to the solution in advance.

[Surfactant] In order to reduce the interfacial tension between the specific binding substance solution and the carrier liquid, a surfactant may be added to the solvent of the specific binding substance. As the surfactant, any of anionic, nonionic, cationic, and fluorine surfactants may be used. In particular,
Sodium dodecylbenzenesulfonate, p-tert
Sodium octylphenoxyethoxyethylsulfonate, saponin, nonylphenoxypolyethoxyethanol, JP-A-62-170950, U.S. Pat.
Fluorinated polymer surfactants described in, for example, 380,644, JP-A-60-244945 and JP-A-6-244945.
Examples thereof include fluorine surfactants described in JP-A-3-188135 and polyalkylene oxides and anionic surfactants described in JP-A-6-301140. The concentration of the surfactant in the solution containing the specific binding substance is 0.001-2%, preferably 0.01-0.5%.
%.

[Salt] Since the droplets of the specific binding substance do not spread on the substrate after the carrier liquid containing the droplets of the specific binding substance is transferred onto the substrate, the carrier liquid containing the droplets of the specific binding substance may be in a solution containing the specific binding substance. Is preferably added with a salt.

As the salt, a water-soluble salt is preferable. Examples of the water-soluble salt include an inorganic salt and an organic salt. Examples of water-soluble salts are listed below, but the invention is not limited thereto. Examples of the cation of the water-soluble salt include alkali metal ions (eg, Li ⁺ , Na ⁺ , Cs ⁺ ) and alkaline earth metal ions (eg, Be ²⁺ , M
g ²⁺ , Ca ²⁺ , Ba ²⁺ ), transition metal ions (eg, Fe ²⁺ , Fe ³⁺ , Ni ²⁺ , Co ²⁺ , Cu ²⁺ ),
NH ⁴⁺ and organic cation ions (eg, quaternary ammonium, aromatic quaternary amine, etc.). As the anion, a halogen ion (for example, F ⁻ , Cl ⁻ , B
r ⁻ , I ⁻ ), phosphate ion, nitrate ion, nitrite ion, sulfate ion, sulfite ion, borate ion, carbonate ion, cyanide ion, complex salts of transition metals (ferricyanic acid, ferrocyanic acid, Ions of organic carboxylic acids (eg, formic acid, acetic acid, propionic acid, benzoic acid, phthalic acid, trichloroacetic acid, salicylic acid, oxalic acid, malonic acid, maleic acid, adipic acid), organic sulfonic acids (Eg, ethylsulfonic acid, benzenesulfonic acid, tosylic acid, etc.) and phenols (eg, phenol, picric acid, etc.). These cations and anions can be arbitrarily combined. Examples of combinations of the above cations and anions include LiCl, NaCl, KC
1, NaBr, KI, CaI, Na ₂ CO ₃ , NaHC
O ₃ , NaH ₂ PO ₄ , NaKHPO ₄ , K ₃ (Fe
(CN) ₆ ), KNO ₃ , (NH ₄ ) ₂ SO ₃ , Na ₂
HBO _{3 and the} like. The solubility of the salt is as follows.
Those which are soluble in water of 0 × 10 ⁻³ mol / l or more are preferable,
More preferably 1 × 10 ^-2 mol / l or more is soluble.
Further, those having strong ionic strength for both cations and anions are more preferable.

The salt is preferably one that maintains the solubility of the specific binding substance, and particularly a combination of salts that expresses an optimal pH buffering ability when reacting with the substrate and fixing the specific binding substance to the substrate surface. . Combinations of pH buffer salts are described in Chemical Handbook, 2nd Edition, Basic Edition II (Chemical Society of Japan) 13
It is described on pages 12-1320. The salt used in the present invention may be used in combination of plural kinds, and when the salt for maintaining the solubility of the specific binding substance and the salt having a pH buffering ability are different, it is preferable to use them in combination.

The aqueous solution of the specific binding substance is, for example, 5 × 1
0 ^-3 M of fourth calf sodium borate aqueous DNA from thymus was dissolved 0.1% by weight, the aerosol OT (manufactured by American Cyanamid Company) as the surfactant in the aqueous solution 0.1 Weight percent, etc.
The present invention is not limited to this aqueous solution.

[Substrate] As the substrate, inorganic materials such as glass and ceramic plate, polymers such as polyolefin and nylon, and other known materials can be used. Alternatively, a composite in which the polymer surface is glass-coated may be used. The substrate preferably has a smooth surface on which the specific binding substance is arranged. The size of one substrate is usually 26 × 78 mm, but the present invention is not limited to this size.

The specific binding substance is arranged in a matrix on the substrate, but is not limited to this arrangement. When a specific binding substance is arranged in a matrix, that is, on each intersection of vertical and horizontal parallel lines, the specific binding substance is substantially circular, and the diameter is preferably 10 to 1000 μm.
m, more preferably 50 to 500 μm. The distance between the specific binding substances in the vertical direction or the horizontal direction is 0.1 to 10 m between the respective center points when the specific binding substance is regarded as a circle.
m, more preferably 0.2 to 2 mm. The number of spots of the specific binding substance provided on one substrate is 100 to
It is 30,000, more preferably 300 to 10,000.

FIG. 1 is a schematic diagram showing a test chip manufacturing apparatus according to the present invention. The test chip manufacturing apparatus 10 includes a substrate holding table 11, a flow tube 13 having a nozzle 12, a carrier liquid supply unit 14, a droplet sending unit 15, a liquid draining unit 16, and a droplet arrangement unit 17. Have been.

The substrate holding table 11 has a clamp (not shown) for holding the glass substrate 20.
Fix it so that it does not shift. The clamp may be a mechanical clamp that holds, holds, or holds the glass substrate 20, or may be a clamp that holds the glass substrate 20 by air suction, electrostatic suction, or the like. This substrate holder 11
Is configured to fix one glass substrate 20 in the present embodiment, but is preferably configured to hold many glass substrates from the viewpoint of improving productivity. Further, a substrate supply unit and a substrate discharge unit (not shown) are connected to the substrate holding table 11, and supply and discharge of the substrate are performed.

The substrate contact portion of the substrate holding table 11 is made of a metal plate, and the metal plate
Are connected, and a DC high voltage is applied. Thus, an electrostatic field is applied to the glass substrate 20 to facilitate the adhesion of the carrier liquid 22 to the glass substrate 20. The electric field strength of the electrostatic field is preferably from 0.01 to 500 kV / m, and more preferably from 0.1 to 100 kV / m.

A flow tube 13 is arranged above the substrate 20 in a vertical direction and the nozzle 12 faces the glass substrate 20. The flow pipe 13 has a carrier liquid supply unit 14
And the droplet sending section 15 are connected. The carrier liquid supply unit 14 includes a tank and a pump (not shown),
The carrier liquid 22 is supplied into the flow pipe 13. The pump is constituted by a plunger pump, and its liquid sending flow rate is 1 to 500 μl / min. It is sufficient that the carrier liquid can be sent out at a constant speed in the above range, and the liquid sending method is not limited to the plunger pump. For example,
A carrier liquid tank may be provided at a high place, and the carrier liquid may be drawn out using the head thereof and the surface tension of the carrier liquid between the nozzle outlet and the substrate.

As shown in FIG. 2, the droplet sending section 15 is configured such that one drop of the carrier liquid 22 in the flow pipe 13 is 0.1 to 200
nl droplets of a solution of a specific binding substance 32 in a small amount are supplied.
A dosing valve 33 for supplying a solution of the specific binding substance 32 to the micro nozzle 31;
And a switching valve 34 for switching the solution No. 2 and supplying the solution to the dosing valve 33. By the droplet sending section 15, droplets of the specific binding substance 32 are arranged at predetermined intervals in the carrier liquid 22 in the flow pipe 13.

The droplet is not limited to the above range, but preferably has a volume of 0.05 to 500 nl. Further, instead of the dosing valve 33, a solution of the specific binding substance may be supplied into the carrier liquid using a small amount of a dispenser or the like. Further, the number of micro nozzles is not limited to one, and a plurality of micro nozzles may be arranged. The arrangement direction may be arranged at an appropriate pitch in the circumferential direction of the flow pipe, or may be arranged at an appropriate pitch in the flow direction of the carrier liquid in the flow pipe. May be arranged. As described above, by increasing the number of the minute nozzles and increasing the switching valve provided for each minute nozzle, it is possible to cope with an increase in the types of the specific binding substances. When the switching valve 34 is used,
It is preferable that the carrier liquid 22 be present between the droplets so that the droplets of the specific binding substances do not mix between the switching valve 34 and the outlet of the micro nozzle 31.

Further, it is preferable that the liquid drop sending section 15 is provided with a flow velocity measuring mechanism 35 for measuring the flow velocity of the carrier liquid 22, and a controller 36 controlled by a computer. The controller 36 controls the opening / closing timing of the dosing valve 33 and the switching valve 34 based on the flow rate signal from the flow rate measuring mechanism 35, and arranges the droplets of the specific binding substance 32 in the carrier liquid 22 at regular intervals.

As shown in FIG. 1, the flow pipe 13 has a plane-direction shift mechanism 41 and a
Is connected to the vertical shift mechanism 42. The plane direction shift mechanism 41 moves the nozzle 12 to the glass substrate 2 when the nozzle 12 is in the retracted position as described later.
By moving the nozzle 12 in a two-dimensional direction parallel to 0, the tip of the nozzle 12 is stopped at the crossing point on the glass substrate 20, and the carrier liquid 22 having the droplet of the specific binding substance 32 is arranged in a matrix. The moving speed of the plane direction shift mechanism 41 is usually 0.1 to 10 mm / sec. The surface direction shift mechanism 41 is composed of an X direction shift mechanism and a Y direction shift mechanism (not shown), and a general shift mechanism is used. In addition, it may be constituted by a robot hand or the like.

The vertical shift mechanism 42 moves the flow pipe 13 in the vertical direction, and as shown in FIG.
(2) A contact position where the carrier liquid 22 at the leading end is brought into contact with the glass substrate 20 (a first position of claim 13), and a retracted position (FIG. 13) 2 position). At the contact position, the distance L1 between the surface of the glass substrate 20 and the tip of the nozzle 12 is:
It is set to 0.1 to 3 mm. In the retreat position, the distance L2 is set to 0.5 to 5 mm. The distance L1 between the substrate surface and the tip of the nozzle at the contact position is 0.2 to 30.
mm is a preferred range. The distance L2 between the substrate surface and the nozzle tip at the retracted position is 0.01 to 10 m.
m is a preferred range.

The plane shift mechanism 41 and the vertical shift mechanism 42 are controlled by a controller (not shown). The controller controls the vertical shift mechanism 42 in synchronization with the discharge of the carrier liquid 22 having the droplet of the specific binding substance 32 from the tip of the nozzle 12, and controls the carrier liquid 22 having the droplet of the specific binding substance 32. Are arranged in a matrix on the glass substrate 20.

FIG. 3 shows this nozzle shift state, where (a), (d) and (e) show the nozzles at the retracted position, and (b) and (c) show the nozzles at the contact position. Respectively. (A) is a state in which the application of the carrier liquid 22 is started, and shows a state where the carrier liquid 22 gradually becomes droplets due to surface tension at the lower end of the nozzle 12 due to the constant supply of the carrier liquid 22. (B)
This shows a state immediately after being located at the contact position. At this time, the increased droplet of the carrier liquid 22 has not yet reached the surface of the glass substrate 20. Eventually, when the carrier liquid 22 increases and comes into contact with the surface of the glass substrate 20, the state shown in FIG. In this state (c), the carrier liquid 22 is continuous between the glass substrate 20 and the nozzle 12. Next, the nozzle 12 is raised by the vertical shift mechanism 42 to the retracted position (d). The liquid is drained by the movement to the retracted position in (d), and the carrier liquid 22 having the droplet of the specific binding substance 32 is transferred from the tip of the nozzle 12 to the glass substrate 20. Thereafter, as shown in (e), the nozzle 12 is positioned at the next carrier liquid application position in the plane direction of the glass substrate 20 by the plane parallel shift mechanism 41. Hereinafter, by repeating the operations (a) to (e), various specific binding substances 32 are formed on the glass substrate 20.
Are arranged as spots 50 in a matrix form.

FIG. 4A shows the spots 50 of the specific binding substance arranged on the glass substrate 20 in this manner. (B) shows a comparative example in which the carrier liquid 51 is continuously flowed on the glass substrate 52 and the spots 53 of the specific binding substance are arranged. When the spot 53 of the specific binding substance 32 is disposed by continuously flowing the carrier liquid 51 of (b), the droplet of the specific binding substance 32 moves in the carrier liquid 51 in the front, rear, left, and right directions. The spots 53 will no longer be arranged in a matrix, making it difficult to position the spots later when the test chip is used for biological analysis.

On the other hand, in the case of the present invention, since the droplets of the carrier liquid 22 containing the specific binding substance 32 of the nozzle 12 are surely positioned at predetermined positions and arranged by the liquid draining section 16, The flow range of the droplet of the specific binding substance in the carrier liquid becomes small, and the spots 50 can be arranged in a matrix with high accuracy.

In the above embodiment, immediately after the contact position, the droplet at the tip of the nozzle is prevented from adhering to the substrate. In addition, as shown in FIG. At the evacuation position, the specific binding substance 32 at the tip of the nozzle 12
May be increased, and the droplet may be brought into contact with the glass substrate 20 at the contact position shown in FIG. 5B. Also in this case, the droplets can be reliably arranged in a matrix on the glass substrate 20,
A test chip with high spot positioning accuracy can be manufactured.

In the above embodiment, the nozzle 12 and the flow pipe 13 are moved by using the plane shift mechanism 41 and the vertical shift mechanism 42, but the plane shift mechanism and the vertical shift mechanism are provided on the substrate holding table 11 side. And the glass substrate 2
Each spot 50 may be arranged in a matrix at 0.
Further, one of the X-direction shift mechanism and the Y-direction shift mechanism of the surface direction shift mechanism 41 may be provided on the nozzle 12 side, and the other may be provided on the substrate holding table 11 side.

In the above embodiment, the operation timing of each shift mechanism is determined in consideration of the flow rate of the carrier liquid. In addition, a droplet of the specific binding substance is placed at the tip of the nozzle or in the middle of the nozzle. Droplet detecting means such as a laser sensor for detecting may be provided, and the operation timing of each shift mechanism may be determined based on a detection signal from the drop detecting means. In this case, since the operation of each shift mechanism is performed after detecting the droplet of the specific binding substance, the droplet of the specific binding substance can be reliably arranged on the substrate.

In the above embodiment, one nozzle is used.
This may be provided in plurality. In this case, the production efficiency is improved. Further, a collecting nozzle for collecting the excess carrier liquid may be attached to the tip of the nozzle 12. This recovery nozzle may be provided integrally with the nozzle 12.

[0044]

As described above, according to the method and apparatus for producing a test chip of the present invention, a liquid containing a specific binding substance is contained in a carrier liquid flowing through a flow pipe having a nozzle at the tip. It is sent intermittently as droplets. When arranging the carrier liquid containing the specific binding substance, the distance between the nozzle tip and the substrate is reduced, and the carrier liquid containing the specific binding substance droplet is brought into contact with the substrate. The specific binding substances can be accurately arranged in a matrix on the substrate by transferring the carrier liquid containing the specific binding substance droplets from the nozzle tip to the substrate by performing liquid drainage by widening the distance between the nozzles. The obtained test chip facilitates the positioning of a specific binding substance in biological analysis.

Further, when arranging the spots, an electrostatic field is applied to the carrier liquid, so that the carrier liquid can be easily attached to the substrate, and a test chip with high positioning accuracy of each spot can be obtained.

Further, the carrier liquid is an organic solvent,
When the solvent of the liquid containing the specific binding substance is water, the specific binding substance in the carrier liquid is transferred onto the substrate without being diffused, so that the number of spots formed can be increased.

[Brief description of the drawings]

FIG. 1 is a schematic diagram of a test chip manufacturing apparatus according to the present invention.

FIG. 2 is a diagram showing one embodiment of a part of the device shown in FIG. 1;

FIG. 3 is a diagram illustrating a method for manufacturing a test chip according to the present invention.

FIG. 4 is a plan view of a test chip manufactured by the manufacturing method shown in FIG.

FIG. 5 is a view for explaining another embodiment of the test chip manufacturing method according to the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 10 Test chip manufacturing apparatus 11 Substrate holding table 12 Nozzle 13 Flow pipe 14 Carrier liquid supply part 15 Droplet sending part 16 Drainage part 17 Droplet arrangement part 20 Glass substrate 21 DC high voltage generation part 22 Carrier liquid 31 Micro nozzle 32 Unique Dynamic binding substance 33 Dosing valve 34 Switching valve 35 Flow velocity measurement mechanism 36 Controller 41 Planar shift mechanism 42 Vertical shift mechanism 50 Spot of specific binding substance

Claims (15)

[Claims] 1. A liquid containing a specific binding substance is intermittently sent as a droplet to a flowing carrier liquid, and the carrier liquid is sent out from a nozzle onto a substrate, and the specific binding substance is put on the substrate. When arranging the carrier liquid containing, after reducing the distance between the nozzle tip and the substrate and bringing the substrate into contact with the carrier liquid containing the specific binding substance droplet, the distance between the nozzle tip and the substrate is increased. Transferring a carrier liquid containing a specific binding substance droplet from a nozzle tip to a substrate by using the method. 2. The method according to claim 1, wherein when arranging the carrier liquid containing the specific binding substance on the substrate, an electrostatic field is applied to the carrier liquid containing the droplet of the specific binding substance. Test chip manufacturing method. 3. A liquid containing a specific binding substance is intermittently sent as a droplet to a flowing carrier liquid, and the carrier liquid is sent out from a nozzle onto a substrate, and the specific binding substance is put on the substrate. A method for producing a test chip, comprising: applying an electrostatic field to a carrier liquid containing droplets of a specific binding substance when arranging a carrier liquid containing the same. 4. The test chip according to claim 1, wherein the carrier liquid is an organic solvent, and the solvent of the liquid containing the specific binding substance is water. Production method. 5. The method according to claim 4, wherein the organic solvent comprises at least one of alkanes, ketones, and esters. 6. The method for producing a test chip according to claim 1, wherein the specific binding substance is any substance having a cDNA function. 7. The method for producing a test chip according to claim 6, wherein the substance having a cDNA function is a substance having a plurality of different types of cDNA functions. 8. The test chip according to claim 1, wherein an interfacial tension between the carrier liquid and the liquid containing the specific binding substance is 1 mN / m or less. Production method. 9. The method for producing a test chip according to claim 1, wherein a surfactant is added to the solution containing the specific binding substance. 10. The method according to claim 9, wherein the surfactant contains at least one of sulfosuccinates and gluconiamides. 11. The method for producing a test chip according to claim 1, wherein a salt is added to the solution containing the specific binding substance. 12. The salt according to claim 1, wherein the cation is an alkali metal ion, and the anion is a salt formed by combining a polyvalent carboxylate ion, a halide ion and an inorganic acid ion, and contains at least one of them. The method for manufacturing a test chip according to claim 11, wherein: 13. A test chip manufacturing apparatus in which a specific binding substance is arranged on a substrate, a pipe through which a carrier liquid flows, means for feeding the carrier liquid into the pipe, and the specific binding connected to the pipe. A liquid droplet sending means for intermittently sending a liquid containing a substance as a liquid droplet into a carrier liquid; a nozzle provided at a tip of the tube; a holding member for holding the substrate; A first position in which the nozzle tip is moved closer to and away from each other to bring the nozzle tip close to the substrate so that the carrier liquid at the nozzle tip comes into contact with the substrate; Liquid draining means for alternately displacing a carrier liquid containing a liquid droplet of a substance from a nozzle tip to a second position for transferring the liquid to the substrate as liquid droplets; The support member is relatively moved in the surface direction of the substrate manufacturing apparatus of a test chip, characterized in that the droplets of the specific binding substance and a sequence unit for arranging the substrate. 14. The test chip manufacturing apparatus according to claim 13, further comprising high voltage applying means for applying a DC high voltage to said holding member. 15. The liquid according to claim 13, wherein the liquid containing the specific binding substance is a plurality of kinds of liquids, and the droplet sending means switches the plurality of kinds of liquids and sends out the liquids sequentially. Test chip manufacturing equipment.