JP3507462B2 - Method for producing probe carrier and apparatus used therefor - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for manufacturing a probe array on a solid substrate and an apparatus used for this method. This device includes a liquid ejection device having a configuration for manufacturing a probe array in which probes are fixed in a two-dimensional array on a solid-phase substrate, and a probe array manufacturing device having the liquid ejection device.

[0002]

2. Description of the Related Art When analyzing a base sequence of a gene DNA or simultaneously performing a gene diagnosis with high reliability for many items, a DNA having a target base sequence is selected by using plural kinds of probes. Will be required. DNA microchips have been attracting attention as a means for providing a plurality of types of probes used for this sorting operation. Also, in high throughput screening of drugs and combinatorial chemistry, a large number of solutions of target proteins and drugs (for example, 96 species, 38
4 kinds, 1536 kinds, etc.) are arranged and it is necessary to carry out an orderly screening. For that purpose, we have developed methods for arranging multiple types of drugs, automated screening technology in that state, dedicated equipment, software for controlling a series of screening operations and statistically processing the results. Has been done.

These parallel screening operations basically consist of a large number of known probes as a means for selecting substances to be evaluated, so-called probe.
By using an array, it is possible to detect the presence or absence of an action or reaction on the probe under the same conditions. In general, what kind of action and reaction to use for the probe is determined in advance. Therefore, the probe species mounted in one probe array may be, for example, a group of DNA probes having different base sequences. It is one type of substance when roughly classified. That is, the substance used for the group of probes is, for example, DNA, protein, or a synthesized chemical substance (drug). In many cases, a probe array composed of a plurality of types of probes is often used. However, depending on the nature of the screening work, as probes, DNA having the same base sequence, protein having the same amino acid sequence, and the same chemical It is also possible to use an array form in which a large number of substances are arranged. These are mainly used for drug screening and the like.

In a probe array consisting of a plurality of types of probes, specifically, a group of DNAs having different base sequences, a group of proteins having different amino acid sequences, or a group of different chemical substances is used. In many cases, a plurality of types of constituents are arranged in an array on a substrate or the like according to a predetermined arrangement order.
Among them, the DNA probe array is used for analysis of the base sequence of gene DNA and, at the same time, for highly reliable gene diagnosis of many items.

One of the problems in a probe array consisting of a plurality of types of probes forming this group is to use as many types of probes as possible, for example, D having various types of base sequences.
The NA probe is placed on one substrate. In other words, how densely the probes can be arranged in an array.

As one method for fixing a plurality of kinds of probes in an array on a substrate, US Pat. No. 5,424,186
A method for producing DNA probes having different base sequences in an array form by a sequential extension reaction of DNA on a solid-phase substrate using a photodegradable protective group and photolithography described in Japanese Patent Publication No. be able to. When this method is used, for example, 10000 per cm ²
It is also possible to manufacture a DNA probe array on which DNAs of different kinds or more are mounted. In this method,
When synthesizing DNA by a sequential extension reaction, a photolithography process is performed for each of the four types of bases (A, T, C, G) using a dedicated photomask, and any base is placed at a predetermined position on the array. Is selectively extended to synthesize a plurality of types of DNA having a desired nucleotide sequence with a predetermined sequence on the substrate. Therefore, when the chain length of DNA is long, the cost required for production is high and it takes a long time. In addition, since the efficiency of nucleotide synthesis in each extension step is not 100%, the ratio of DNA in which the designed nucleotide sequence has a defect is not small. Furthermore, in the case of using a photodegradable protective group in the synthesis, the synthesis efficiency is lower than in the case of using an ordinary acid-decomposable protective group,
In the finally obtained array, there is also a problem that the proportion of DNA according to the designed nucleotide sequence becomes small.

Further, since the product directly synthesized on the solid phase substrate is used as it is, it is of course impossible to remove the DNA having a defective base sequence from the designed DNA according to the purified base by purification fractionation. Is. In addition, in the finally obtained array, there is a problem that the base sequence of the DNA synthesized on the substrate cannot be confirmed. This is because if a given base was hardly extended at a certain extension stage due to an error in the process, and it was a completely defective product, the screening using this defective product probe array resulted in an incorrect result. However, it means that there is no way to prevent it. The inability to confirm this nucleotide sequence is the biggest and essential problem in this method.

As a method different from the above method, DNA for a probe is previously synthesized and purified, and in some cases, the base length thereof is confirmed, and then each DNA is supplied onto a substrate by a device such as a microdispenser. And probe
Techniques for making arrays have also been proposed. PCT publication WO95 / 355O5 describes a method of supplying DNA onto a membrane using a capillary. By applying this method, it is possible in principle to manufacture about 1000 DNA arrays per cm ² . Basically, one capillary dispense device for each probe is used to supply the probe solution to a predetermined position on the substrate, and the operation is repeated to make the probe solution
This is a method of manufacturing an array. There is no problem if you prepare a dedicated capillary for each probe, but if you try to do the same work with a small number of capillaries, to prevent cross-contamination, the capillaries should be sufficient when replacing the probe type. Need to be washed. Further, it is necessary to control the supply position each time. Therefore, it cannot be said that the method is suitable for manufacturing an array in which many kinds of probes are arranged at high density. In addition, since the probe solution is supplied to the substrate by tapping the tip of the capillary onto the substrate, reproducibility and reliability cannot be said to be perfect.

[0009] In addition, 96 wells or 384 used for high-throughput screening of drugs.
Micro-dispenser devices are also commercially available, for example from Robbins Scientific, Inc. under the trade name of HYDRA ^™ , for supplying different drug solutions to a well microplate in individual wells. This is basically a two-dimensional array of microsyringes, and the minimum discharge amount is 100 nl. If this is applied to the array formation, the density is limited by this minimum ejection amount, and there is a limit to high density.

Another method is to use DN on the substrate.
When performing the solid phase synthesis of A, a method of supplying a solution of a substance required for the synthesis onto the substrate by an inkjet method at each extension step has also been proposed. For example, European Patent Publication EP 0
703 825B1 is a DNA having a predetermined nucleotide sequence, which is used in solid phase synthesis of DNA, by supplying a nucleotide monomer and an activator from different piezo jet nozzles.
Methods for solid phase synthesis of multiple species have been described. The supply (coating) by the ink jet method is more reliable than the supply (coating) of the solution using the capillary, such as the reproducibility of the supply amount, and the structure of the nozzle can be miniaturized. It has characteristics suitable for increasing the density of probe arrays. However, since this method also basically applies the sequential extension reaction of DNA on the substrate, it is the greatest problem in the method described in the above-mentioned US Pat. No. 5,424,186. There still remain problems such as the inability to confirm the base sequence of the DNA synthesized on the substrate. Although the complexity of performing a photolithography process using a dedicated mask for each extension step is eliminated,
It involves some problems in that a given probe is fixed at each point, which is an essential requirement of the array. Incidentally, the EP 0,703,825B1 describes only a method of using a plurality of piezo jet nozzles formed independently, and when using this small number of nozzles, a method of using the above-mentioned capillary. alike,
It is not always suitable for high density probe array manufacturing.

Further, Japanese Patent Application Laid-Open No. 11-187900 discloses a method in which a liquid containing a probe is attached to a solid phase as a liquid by a thermal ink jet head to form spots containing the probe on the solid phase. . In this method, an inkjet head for a general printer is used as an apparatus for ejecting liquid, but the structure of the inkjet head for this printer may not be optimal for manufacturing the probe array.

The inkjet head for printing has been developed for printing characters and images. Therefore, the liquid used is one color (black) ink for monochrome (generally black) printing, and is generally the three primary colors of color for color printing, that is, yellow (Y), cyan ( Ink of three colors of C) and magenta (M). In case of color printing, black or Y, M,
There are cases where the dark and light inks of C and C are used, but at most 1
Never use more than one type of ink.

Further, since a large amount of ink is used for printing on paper, a conventional ink jet printing head has a tank (reservoir) for filling ink having a sufficient capacity for printing, and an ink. A flow path for guiding the ink to the nozzle and a nozzle for ejecting ink are provided.

On the other hand, the liquid ejecting apparatus used for ejecting the liquid at the time of manufacturing the probe array is desired to eject as many kinds of liquid as possible as described above. When there is a liquid ejecting apparatus having a plurality of nozzles, a liquid ejecting apparatus having a liquid reservoir that has a one-to-one correspondence with the plurality of nozzles and the plurality of nozzles is desirable.

Further, in the production of the probe array by the liquid ejecting apparatus for producing the probe array, since the liquid is not consumed as much as when printing is performed on the paper surface by the conventional general inkjet head for printing,
A relatively small volume of the reservoir of the liquid ejection device is sufficient.

Further, in a general ink jet head for printing, it is necessary to eject a desired ink to a desired position on a paper surface in order to form a character or an image. Therefore, each nozzle is independently operated at an arbitrary timing. The head configuration is selectable with. As a result, the head has a complicated structure.

On the other hand, in the liquid ejecting apparatus used for ejecting the liquid at the time of manufacturing the probe array, it is not always necessary to select each nozzle independently at any timing.

As described above, the conventional general inkjet head for printing has a head configuration in which each nozzle can be independently selected at any timing. At this time, ink is ejected from a desired nozzle. The power transistor and the logic circuit necessary for this purpose may be provided outside the head or inside the head.

By the way, as a method for ejecting the liquid, a thermal jet method for ejecting the liquid by thermal energy generated from a heater, and a piezo jet for ejecting the liquid by the deformation of the element caused by applying a voltage to the piezo element. There is a method. Among these, the thermal jet method has a simpler structure than the piezo jet method, and is suitable for downsizing the head and increasing the number of nozzles. From this point, the thermal jet method is considered to be suitable when a large number of nozzles are desired for the liquid ejection apparatus for manufacturing the probe array.

Some of the current thermal jet printing heads have a large number of nozzles, for example, 128 or 256, for each color in order to improve the printing speed. As the number of nozzles increases, the number of signals for determining whether or not ejection is performed in each nozzle also increases, so that the number of contact points between the head and the outside increases. In order to solve this problem, a power transistor for driving the heater, a shift register for transferring the data of characters and images to be printed, and circuits such as a decoder, an AND, and a NAND are mounted in the head to connect the head and the outside. The number of contacts of is decreasing.

Further, since the power transistor and the logic circuit are mounted on the head, silicon is generally used as a material forming the head, and elements such as a MOS transistor and a bipolar transistor are formed in the silicon.

As described above, in the conventional printing head, a semiconductor element is generally built in the head. In such a case, the head manufacturing process becomes complicated and complicated. Due to such a process, problems such as an increase in head cost and a decrease in yield may occur. In particular, the decrease in yield often becomes a problem as the number of nozzles increases, that is, the number of circuits formed in the head increases.

As described above, the liquid ejecting apparatus for producing the probe array has a large number of nozzles, for example, 10 nozzles.
Since a structure having 00 or more nozzles is desired, problems such as cost increase and yield reduction cannot be avoided by simply applying the structure of the conventional inkjet head.

Further, in the case of manufacturing a probe array in which different liquids are arranged by discharging different liquids from the respective nozzles, when manufacturing one probe array, the discharge from each nozzle is In many cases, one time is sufficient, and the position where the liquid is placed is also predetermined. That is, unlike the case of printing on paper, the probe array manufacturing by this method does not necessarily require complicated discharge control as described above, and therefore has a simple structure, low cost, and high yield. A liquid ejection device is desired.

[0025]

As described above, according to the conventional method for manufacturing a probe array, an array in which a large number of kinds of probes are mounted on a substrate at a high density can be reproducibly and simply manufactured. In many cases, some problems remain in terms of manufacturing. Therefore, for example, a new method useful in manufacturing a two-dimensional array array in which the base sequences of many types of DNA probes are confirmed in advance and then arranged at a high density according to a predetermined sequence order. Is suggested.

The present invention solves such a problem, and an object of the present invention is to obtain a plurality of highly pure probes of a high purity in a predetermined sequence order, in which unnecessary components other than the desired probe are eliminated as much as possible. It is an object of the present invention to provide a method of manufacturing a probe array capable of improving reproducibility and workability when manufacturing a two-dimensional array array arranged at a high density. Another object of the present invention is to provide a liquid ejecting apparatus having a configuration extremely suitable for carrying out this manufacturing method, a probe array manufacturing apparatus using the same, and a liquid ejecting apparatus and a probe array using the same. To provide a manufacturing apparatus.

More specifically, a probe that makes it possible to manufacture various types of high-density probe arrays that are different from each other with higher reproducibility and easily on a large number of substrates. -To provide an array manufacturing method and various devices used for the method.

[0028]

Means for Solving the Problems The inventors of the present invention have earnestly studied to solve the above problems, and have obtained the following findings.

(1) A plurality of types of probes to be used themselves are separately manufactured, purified, etc., and, if necessary, identified and confirmed to be the target probe, and a probe containing the probe of high purity. A two-dimensional probe array having a high density can be obtained by applying a solution as a minute liquid having a predetermined liquid amount on a substrate and arranging the solution in an array.

(2) As means for applying the probe solution used in the operation in the above (1) to the substrate as a minute liquid having a predetermined liquid amount, it is connected to a solution reservoir for accommodating each probe solution and it. By adopting a liquid ejection device having a number of solution ejection nozzles corresponding to a plurality of types of probes to be applied, the liquid amount of the probe solution ejected from each nozzle and applied on the substrate is
High reproducibility is obtained for each nozzle, and
The uniformity between nozzles can also be increased. Also, this configuration
It is possible to omit the conventional step of replacing the probe solution in the ejection device, which has been present when a large number of types of probe solutions are applied to the carrier using the ejection device. As a result, the step of cleaning the ejection device when replacing the probe solution can be omitted, and there is no risk of contamination with the probe solution before replacement, which simplifies the manufacturing process and improves the reliability of the resulting probe carrier. Contribute to the improvement of.

(3) Since the liquid ejecting apparatus itself to be used can have a shape in which a plurality of nozzles are arranged in an array with a sufficiently high density, as a result, a plurality of types of desired high-purity probes are provided on the same substrate. It becomes possible to coat the substrate in a two-dimensional array having a desired high density with extremely high reproducibility and without complicated work.

(4) When a plurality of nozzles suitable for the above-mentioned use are assembled in the same liquid ejecting apparatus, a solution reservoir for accommodating each probe solution and its connection are connected according to the order of arrangement in the two-dimensional probe array formed By disposing the solution discharge nozzle at a predetermined position (sequence), a plurality of two-dimensional probe arrays having the same probe arrangement can be provided.
It is possible to repeatedly manufacture the liquid ejecting apparatus as a whole, without performing positioning adjustment of the ejection positions from the individual nozzles, and only by slightly adjusting the positioning.

(5) As means for applying the probe solution in the operation in the above (1) as a minute liquid having a predetermined liquid amount on the substrate, a liquid reservoir for storing each probe solution and a liquid discharge connected thereto Nozzle
It is equipped with the number corresponding to multiple types of probes to be applied,
By preferably adopting a liquid ejection device formed in the shape of a flat chip, the liquid amount of the probe solution ejected from each nozzle and applied on the substrate can be highly reproducible in each nozzle, and The uniformity between nozzles can also be increased.

(6) Since the above-described liquid ejecting apparatus can have a shape in which a plurality of nozzles are arranged in an array with a sufficiently high density, as a result, a plurality of types of desired high-purity probes can be provided on the same substrate. It is possible to apply onto a substrate in a two-dimensional array having a desired high density with reproducibility and without complicated work.

The present inventors have completed the present invention mainly based on the above findings.

That is, a liquid ejecting apparatus according to one aspect of the present invention that can achieve the above object is a liquid ejecting apparatus for producing a probe carrier having a plurality of types of probes on different positions on a carrier. In addition, a liquid reservoir (solution reservoir) for storing a liquid containing the probe (probe solution) and a liquid ejection nozzle (solution ejection nozzle) communicating with the liquid reservoir are provided at least in a number corresponding to the plurality of types of probes. , The reservoir and the liquid discharge nozzle are formed on the same substrate,
In addition, the reservoir is a liquid ejecting apparatus characterized in that each of the reservoirs has a supply port for supplying a different solution.

It is preferable that the probe is at least a part of a nucleic acid.

The plurality of liquid discharge nozzles may be linear or may be opened in the vertical and horizontal directions. It is preferable that the liquid reservoir is located on the opposite side of the liquid discharging nozzle from the liquid discharging direction. It is more preferable that a separate liquid storage portion is connected to and connected to the liquid reservoir. The liquid in the nozzle is preferably provided with a thermal energy generator for applying thermal energy to eject the liquid, and bubbles generated by applying thermal energy to the liquid communicate with the outside air. It is more preferable that the liquid ejection device ejects the liquid. The liquid ejecting apparatus is an apparatus for applying the liquid onto the carrier at the same density as the nozzle density of the liquid ejecting apparatus, or the liquid ejecting apparatus uses one of the relative movements of the carrier. More preferably, it is a device for applying the liquid containing the probe to the carrier so as to have a density higher than the nozzle density of the liquid ejection device by performing the process a plurality of times on the carrier.

A liquid ejecting apparatus according to an aspect of the present invention includes an integrated flat plate-shaped substrate having a laminated layer structure,
A plurality of liquid ejection nozzles formed and arranged in a two-dimensional array on one surface of the substrate, and a two-dimensional array formed on the other surface of the substrate by individually communicating with the plurality of liquid ejection nozzles A liquid reservoir arranged, a liquid discharge energy generating element formed and arranged in a two-dimensional array corresponding to each of the liquid discharge nozzles, and provided with an opening on the surface on which the liquid reservoir is arranged. And a supply port for supplying a liquid to each of the reservoirs. Further, a probe carrier manufacturing apparatus including the above liquid ejecting apparatus is also included in the present invention.

An apparatus for producing a probe carrier according to an aspect of the present invention is an apparatus for producing a probe carrier having a plurality of types of probes on different positions on a carrier, and stores a liquid (probe solution) containing the probe. A plurality of liquid reservoirs (solution reservoirs) and liquid discharge nozzles (solution discharge nozzles) communicating therewith corresponding to at least the plurality of types of probes, and the reservoirs and the liquid discharge nozzles are formed on the same substrate. And the reservoir is provided with a liquid ejection device each having a supply port for supplying a different solution, and a positioning for relatively aligning the plurality of liquid ejection nozzles with the carrier. And a means for manufacturing the probe carrier.

A method of manufacturing a probe carrier according to one aspect of the present invention is a method of manufacturing a probe carrier having a plurality of types of probes on different positions on a carrier, which is a liquid reservoir for storing a liquid containing the probe. And at least a number of liquid discharge nozzles communicating therewith corresponding to the plurality of types of probes, the reservoir and the liquid discharge nozzle are formed on the same substrate, and the reservoir stores different liquids. A step of preparing a liquid ejection device each having a supply port for supplying, a step of relatively aligning the plurality of liquid ejection nozzles with the carrier, and a step of moving the liquid ejection nozzle onto the carrier. And a step of ejecting a liquid containing each of the probes to different positions of the probe carrier. is there.

The probe is preferably at least a part of nucleic acid. In the liquid ejecting apparatus, the plurality of liquid ejecting nozzles may be linear or may be opened in the vertical and horizontal directions. It is preferable that the liquid reservoir is located on the opposite side of the liquid discharging nozzle from the liquid discharging direction. It is preferable that a separate liquid storage portion is connected to and connected to the liquid reservoir. The liquid ejecting apparatus applies liquid to the nozzle,
It is preferable to provide a thermal energy generator for applying thermal energy to eject the liquid, and bubbles are generated by applying thermal energy to the liquid to communicate with the outside air, so that the liquid is ejected. More preferably, it is a discharge device. The liquid ejecting apparatus is used to apply the liquid onto a carrier at the same density as the nozzle density of the liquid ejecting apparatus, or relative movement of the liquid ejecting apparatus and the carrier is performed a plurality of times for one carrier. By carrying out, the liquid containing the probe may be applied onto the carrier so as to have a density higher than the nozzle density of the liquid ejecting apparatus.

Among the probe carriers, those in which the probes are arranged in an array are particularly referred to as a probe array in this specification.

A method of manufacturing a probe array according to one aspect of the present invention is a method of manufacturing a probe array in which each of a plurality of types of probes is fixed in a two-dimensional array on a solid-phase substrate. Immobilization of a plurality of types of probes arranged on the solid-phase substrate is performed by using a probe solution containing each probe, a solution reservoir for accommodating each probe solution, and a solution ejection nozzle connected to the solution reservoir. The two-dimensional array is prepared by ejecting the probe solution contained in the solution reservoir from the solution ejection nozzle as a predetermined amount of liquid onto the solid-phase substrate using a liquid ejection device provided in a number corresponding to the probe. Step of applying in a shape,
Immobilizing each probe contained in a plurality of kinds of probe solutions applied in a two-dimensional array on a solid-phase substrate. In this method, it is desirable to use a liquid ejecting apparatus in which the plurality of nozzles constituting the liquid ejecting apparatus are arranged in a one-dimensional array or a two-dimensional array. In addition, a liquid ejecting apparatus is used in which the plurality of nozzles arranged in a one-dimensional array or a two-dimensional array are integrally formed in a one-dimensional array or a two-dimensional array. Is more preferable.

At that time, the solution discharge direction of the nozzles integrally formed in an array form is the Z direction of the liquid discharge device, and the nozzles are provided on the side opposite to the side where the nozzles are formed. It is possible to use a liquid ejection device in which a solution reservoir provided corresponding to is formed.
For example, the nozzles, which are integrally formed in an array, are integrally formed on one surface of the lithographic substrate, and the corresponding solution reservoir is also located on the other surface of the lithographic substrate. It is possible to use a liquid discharge device formed integrally.

In addition, if necessary, it is possible to use a liquid ejecting apparatus which is connected to the integrally formed solution reservoir and further includes a solution storage portion for increasing the volume. In this aspect, the solution storage portion for increasing the volume connected to the solution reservoir is integrally formed in an array shape, and the array shape is formed corresponding to the nozzle array block integrally formed. It is preferable to use a liquid ejection device in which the array-shaped solution storage portions for increasing the volume that are integrally formed with each other are arranged in a stacked structure.

Alternatively, according to the method of the present invention, a plurality of nozzle array block units having a one-dimensional array arrangement are combined to provide a liquid ejection apparatus having a nozzle array block having a two-dimensional array arrangement. It can also be used.

On the other hand, as for the ink jet head itself to be used, a liquid ejecting apparatus in which ink jet is performed by a piezo element can be used. It is also possible to use a so-called thermal jet type liquid discharge device in which inkjet is performed by a heater element. It should be noted that the thermal jet method is currently more suitable for forming higher density nozzles in the same inkjet head.

In the method of the present invention, when the number of kinds of probes in the target probe array is implemented in such a manner that it is necessary to use a liquid ejecting apparatus having a pair of solution reservoirs and nozzles of 100 or more, the present invention is realized. The effect of is more clear and desirable. Preferably, a pair of solution reservoirs and nozzles are used in a mode in which a liquid ejection device having 1000 or more pairs is used. More preferably, the pair of solution reservoir and nozzle is 1000
It is carried out in a mode in which a liquid ejection device including zero or more sets is used. Furthermore, the method of the present invention can be carried out in a mode in which a liquid ejection device having 100,000 or more pairs of solution reservoirs and nozzles is used.

When using an ink jet head having such a large number of nozzles, it is desirable that the area density of the nozzles themselves be equal to the density of the desired probe array in consideration of the above-mentioned ejection positioning. For example, 100 / cm ² or more, 1000 / cm ² or more,
By using an inkjet head having nozzles having a density of 10,000 / cm ² or more and 100,000 / cm ² or more, it is possible to perform ejection positioning of individual nozzles,
It becomes possible to eject the probe solution in an array form at a correspondingly high density. However, in the ink jet head in which the nozzles are provided with high density, whether or not the solution storage section is mounted in addition to the solution reservoir is selected according to the required probe liquid amount. Furthermore, it depends on the structure of the inkjet head and the method of supplying the probe solution to the solution reservoir.

It is also possible to manufacture a probe array having a desired density by using an inkjet head having nozzles at a density lower than that of the desired probe array. However, in this case, it is necessary to appropriately adjust the ejection timing, the pattern and the like according to the desired density and pattern of the probe array.

In addition, a liquid ejecting apparatus according to an embodiment of the present invention that can achieve the above-mentioned object is a probe device in which a plurality of kinds of probes are fixed on a solid-phase substrate in a two-dimensional array arrangement. A liquid ejecting apparatus for ejecting a probe solution for manufacturing an array, comprising a solution reservoir for accommodating each probe solution and a solution ejecting nozzle connected to the reservoir, in a number corresponding to the plurality of types of probes. Is characterized by.

In the probe array manufacturing apparatus according to one embodiment of the present invention, which can achieve the above object, a plurality of types of probes are fixed in a two-dimensional array on a solid-phase substrate. A probe array manufacturing apparatus, wherein each probe is provided as a means for applying each of a plurality of types of probe solutions containing each of the plurality of types of probes in a two-dimensional array on the solid phase substrate. The liquid ejecting apparatus is characterized by comprising a solution reservoir for accommodating the solution and a solution ejecting nozzle connected thereto, the number corresponding to the plurality of kinds of probe solutions.

The liquid ejecting apparatus and the probe array manufacturing apparatus of the present invention are essentially the same as the above-described probe array manufacturing method of the present invention.
A liquid ejection device used exclusively for the purpose of manufacturing an array,
And a probe array manufacturing apparatus. Therefore,
According to various aspects of the above-described probe array manufacturing method, the liquid ejecting apparatus and the manufacturing apparatus may be the liquid ejecting apparatus and the manufacturing apparatus to which corresponding structural features are added. desirable.

A probe according to another embodiment of the present invention
The array manufacturing method is a method of manufacturing a probe array in which each of a plurality of types of probes is fixed in a two-dimensional array on a solid-phase substrate, and a liquid reservoir for storing a liquid and a liquid discharge The number of liquid ejecting sections corresponding to the plurality of types of probes, and the number of the liquid ejecting sections having the ejection energy generating means for ejecting the liquid supplied from the liquid reservoir to the nozzle as the liquid from the nozzle. Are arranged in a two-dimensional array, each of the plurality of types of probe solutions is stored in each liquid reservoir and supplied to the corresponding nozzle, and in that state, As a liquid, it is ejected onto the solid-phase substrate from the nozzles necessary for forming a two-dimensional array arrangement to form a probe solution coating area arranged in a two-dimensional array. A step, in which each probe included in the probe solution constituting each coating region, characterized in that it comprises a step for fixing on a solid substrate.

In this method, it is desirable to use a liquid ejection device in which the plurality of nozzles constituting the liquid ejection device are arranged in a two-dimensional array.

At that time, the liquid ejection direction of the nozzle is perpendicular to the plane formed by the nozzles arranged in the two-dimensional array of the liquid ejection device, and the side opposite to the side where the nozzle is formed. Further, it is possible to use a liquid ejection device in which a liquid reservoir provided corresponding to the nozzle is formed.

In this embodiment, a liquid reservoir for increasing the volume connected to the liquid reservoir is a plate integrally formed in an array, and the plate is arranged in a two-dimensional array of nozzles. Overlaid on the back surface of the substrate of the liquid ejection device provided with a reservoir,
It is possible to use a liquid ejection device in which a liquid reservoir and a volume increasing reservoir are connected to each other.

On the other hand, regarding the liquid ejecting apparatus itself used, it is preferable to use a so-called thermal jet system in which the liquid is ejected by the heater element.

Further, the structure for driving the heater in the liquid ejecting apparatus is composed of only the heater, the wiring such as aluminum for applying a voltage to the heater, and the pad connecting the liquid ejecting apparatus and the outside. A simple one is preferable. In addition, it is desirable that this manufacturing apparatus is also an apparatus to which corresponding structural features are added according to the various aspects described above.

In the present invention, the amount of the probe solution to be applied is appropriately selected in consideration of the areal density of the probe array to be manufactured. At that time, a predetermined amount of liquid is supplied from the solution ejection nozzle. The amount of the probe solution to be discharged can be selected in the range of 0.1 picoliter to 100 picoliter. Similarly, according to the amount of the probe solution to be applied, the application area occupied by the probe solution is appropriately selected. At that time, the two-dimensional probe array to be produced is The occupied area is 0.01 (for example, 0.1 μm × 0.1
μm) μm ^{2 to} 40,000 (for example, 200 μm × 200
(μm) μm ² may be selected.

The probe immobilized on the solid-phase substrate is a molecule that can be recognized by a specific target and is often a surface-immobilized molecule called a ligand. Further, the probe includes an oligonucleotide or a polynucleotide that can be recognized by a specific target, or another polymer. Depending on the context, the term "probe" refers to molecules having a probe function, such as individual polynucleotide molecules, and populations of molecules having the same probe function, such as polynucleotides of the same sequence surface-immobilized in dispersed locations. Say both. Probes and targets are often used interchangeably depending on the context, and the probes are those that are capable of binding to, or become capable of binding, the target as part of a ligand-antiligand pair. The probes and targets in the present invention may include bases as found in nature, or analogs thereof.

Further, as an example of the probe supported on the solid phase substrate, a fluorescent dye and a solid phase substrate are provided on a part of an oligonucleotide having a base sequence capable of hybridizing with a target nucleic acid via a linker. And a compound having a structure in which the compound is bound to the surface of the solid phase substrate at the bonding portion with the solid phase substrate. The position in the molecule of the oligonucleotide of the binding portion between the fluorescent dye and the solid substrate in the case of such a configuration is
It is not particularly limited as long as it does not impair the desired hybridization reaction.

The probe employed in the probe array to which the method of the present invention is applied is appropriately selected according to the purpose of use, but in order to suitably carry out the method of the present invention, it is manufactured. In the two-dimensional probe array, the probes are DNA, RNA, cDNA (complementary DNA), PNA, oligonucleotide,
At least selected from polynucleotides, other nucleic acids, oligopeptides, polypeptides, proteins, enzymes, substrates for enzymes, antibodies, epitopes for antibodies, antigens, hormones, hormone receptors, ligands, ligand receptors, oligosaccharides and polysaccharides It is preferably one type.

On the other hand, the probe array produced by the method of the present invention preferably contains probes having a structure capable of binding to the surface of the solid substrate, and the probes are not immobilized on the solid substrate. It is desirable that the probe is bonded to the surface of the solid phase substrate.

At this time, the structure of the probe capable of binding to the surface of the solid phase substrate is amino group, mercapto group, carboxyl group, hydroxyl group, acid halide (haloformyl group; -COX), halide (-X), Aziridine,
It is formed by introducing at least one organic functional group such as maleimide group, succinimide, isothiocyanate, sulfonyl chloride (-SO ₂ Cl), aldehyde (formyl group; -CHO), hydrazine and iodoacetamide. Preferably there is. Further, the surface of the solid-phase substrate may be subjected to necessary treatment depending on the structure required for binding to the solid-phase substrate on the probe side.

[0067]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, each aspect of the present invention will be described with reference to specific examples.

1) Aspect Example Using Liquid Ejection Device In the first embodiment of the method for producing a probe array according to the present invention, when the two-dimensional probe array is produced, a probe prepared separately beforehand as a solution is used as a substrate. By using a liquid ejecting apparatus in which nozzles are arranged in an array on a surface in a predetermined arrangement, each probe is ejected and applied in a desired liquid amount, thereby achieving high density of various types.

First, an inkjet head configured in an array used in the steps of discharging and applying the probe solution will be described.

Ejection of a solution by ink jet is originally a method developed for ink jet printing (printing). Specifically, the solution used is one color (black) ink for monochrome (generally black) printing, and generally three primary colors for color printing.
That is, three color inks of yellow (Y), cyan (C), and magenta (M) are used. In the case of color printing, black or Y, M, C dark and light inks may be used if necessary, but at most 10 or more kinds of inks will not be used.

Further, since a large amount of ink is used in the conventional inkjet head block for inkjet printing, a tank for filling the ink having a sufficient capacity, a flow path for guiding the ink to the nozzle,
Although the nozzles for ejecting ink are provided, in the case of printing, in order to maintain the fineness of printing, the diameter of each ink dot is maintained even though the ink is often supplied to a large area at a time. Since it is small in itself, a plurality of colors are used for each color in order to maintain the drawing speed within the desired range.
For example, it is configured to include 64, 128, and 256 nozzles. That is, the ink of each color is ejected from the tank through the flow path by a plurality of nozzles.

Next, the structure of the ink jet head will be described.
The thermal jet method, which is one of the inkjet methods, will be taken as an example and described in more detail. Regarding the structure of the inkjet head described below, Japanese Patent Laid-Open No. 06-040
Further detailed description is given in Japanese Patent Laid-Open No. 037, and by referring to it, it is possible to manufacture an inkjet head having such a structure, and a control electric circuit or the like required to operate the inkjet head.

FIGS. 1 and 2 show the structure of a typical ink jet head manufactured by Canon Inc. and Hewlett Packard Inc., which is one of the ink jet methods, that is, a thermal jet, that is, a so-called bubble jet (BJ) method. It is a schematic perspective view which shows an example. As in the structure shown in FIG. 1, the conventional BJ head 1 is
It is formed by the BJ semiconductor chip device 2 that abuts the laser-etched cap 3. In this structure, the cap 3 has the head 1 through the ink inlet 4.
It serves as a guide for the inward flow (indicated by the arrow in the figure) to and for the outward ejection from the head 1 via the plurality of nozzles 5.

The nozzle 5 is formed in the cap 3 as an end release channel. On the BJ chip 2, one or more (for example, 64) heater elements (not shown) are arranged according to the number of nozzles. When energy is applied to the heater element, bubbles of the evaporated ink solvent are formed in the corresponding channels, and the ink is ejected from each nozzle 5 as the volume increases due to the vaporization.

The BJ chip 2 also has a semiconductor diode matrix (not shown), which serves to energize the heater elements adjacent to the channels.

Next, as shown in FIG.
In the Packard thermal jet head 10, ink enters the cap 12 through an inlet 13 located on the side of the cap 12, the cap 12 having nozzle rows 14 arranged orthogonal to the inlet 13. . Ink exits through the surface of cap 12. A flat plate heater 15 is arranged below each nozzle 14. Thus, ink is ejected from the inlet / outlet channel 13 to the nozzle.

The ink jet head having the structure shown in FIGS. 1 and 2 is originally designed so as to be suitable for the case where only about 10 kinds of ink are used. When considering the ejection of array solution, at least 100 probe species
It is not suitable for direct application to the manufacture of probe arrays that require more than one species. It is also possible to use a plurality of inkjet heads having this type of configuration, apply the probe solution to a part of the array with each head, and repeat this operation to complete the entire array. . In that case, it is necessary to replace the head, or it is necessary to perform accurate positioning for each head.

Further, as described above, such a conventional ink jet printer head is for printing and has a large number of nozzles for one type of ink. The structure is not particularly necessary for the manufacture of probe arrays. On the contrary, if you try to configure the probe solution tank according to the required number of nozzles corresponding to the type of probe solution required for manufacturing the probe array, the conventional inkjet head configuration is different for each color ink. In the cap part where multiple nozzles are provided,
In order to arrange one large capacity tank for each color ink,
Due to the two-part structure in which both are formed separately and connected by the ink flow path to form the head, and due to this, as shown in FIGS. 1 and 2, the ink inflow direction and the ink ejection Since the configuration in which the directions are orthogonal to each other is adopted, it is extremely difficult to provide the required number of ink flow paths. That is, as a result, the inkjet head having the two-part structure suitable for the conventional multi-color printer, as it is, causes great difficulty in manufacturing the probe array.

Japanese Unexamined Patent Publication No. 06-040037 describes an integrated BJ head as a means for replacing the ink jet head having the two-part structure. The ZBJ head described here has a structure in which a nozzle part and a reservoir part that holds ink are integrated, instead of the conventional structure using two parts, but this structure is designed for its original purpose of use. The structure is such that it has multiple nozzles in one kind of ink (that is, probe solution). In the present invention, unlike the inkjet heads for these multicolor printers, the configuration of a liquid ejecting apparatus that corresponds to each probe solution and includes a solution reservoir and an individual probe solution ejecting nozzle connected to the reservoir is used. To do. Therefore, the probe solutions ejected from the respective nozzles onto the solid-phase substrate contain different probes, which provides a convenient means for producing a two-dimensional probe array.

The integrated BJ head used in the present invention has a structure in which a solution reservoir and an individual nozzle for discharging a probe solution connected to the reservoir are provided corresponding to each probe solution. And the structure of the individual probe solution discharge nozzle portion itself connected to the reservoir thereof is essentially the same as that disclosed in Japanese Unexamined Patent Publication No.
It can be manufactured according to the design method and manufacturing process of the integrated BJ head described in No. 040037.

In the following description, the term "Z-axis bubble jet chip (ZBJ chip)" means X
It is used to describe a bubble jet tip that lies in the Y plane and ink flows in and out of the tip in the Z direction. Furthermore, for the purpose of helping understanding of the configuration and operation of the liquid ejecting apparatus described, in place of the application example to a plurality of types of probe solutions in the present invention, an operation using an aqueous multicolor ink is performed. The features of the configuration of the liquid ejection device will be described.

FIG. 3 shows a schematic shape of the bubble jet (hereinafter abbreviated as ZBJ) chip 40 arranged in the Z-axis. The ZBJ chip 40 is composed of an ink introducing port arranged on the plane of the chip (on the lower side of the figure) and a plurality of nozzles having ink discharging ports on the opposite side. Figure 1,
The bubble jet chip shown in FIG. 2 has a two-part structure, whereas the ZBJ chip 40 of FIG. 3 is characterized by having a structure formed by a single monolithic integration. This chip 40 can be formed using a semiconductor manufacturing technique. Furthermore, the ink
The ink is ejected from the nozzle 41 in the same direction as the ink is supplied to the chip 40.

Referring now to FIG. 4, there is shown a cross section of an example of a stationary (ie, non-moving) ZBJ printhead 50, which printhead is 1600 dpi.
(Dot / inch) or 400 pixels / inch is formed so that a continuous gray-scale image with an image density of A4 size is created. The head 50 has four nozzle arrays, that is, one ZB having four nozzle arrays indicated by the numbers 71, 72, 73 and 74 in the drawing.
A J chip 70 is provided.

The nozzle arrays 71 to 74 have four pixels per pixel.
It is formed from a nozzle path (via) 77 having one nozzle, giving a total of 51,200 nozzles per chip 70. The enlarged portion of FIG. 4 shows a basic nozzle cross section formed on a silicon substrate 76, on which a layer 78 of thermal SiO ₂ (silicon dioxide) is formed. The heater element 79 is provided around the nozzle 77, and the element 79 is covered with an overcoat layer 80 made of glass by chemical vapor deposition (CVD). Each of the nozzles 77 communicates with a common ink supply channel 75 for a particular color ink. The ZBJ tip 70 can be arranged on the channel bulge 60, and the channel bulge communicates with the common ink supply channel 75 so as to continuously supply the ink flow to the tip 70. Have. A single membrane filter 54 is disposed between the bulge 60 and the tip 70.

Two power supply bus bars 51 and 52 are provided so as to be electrically connected to the chip 70. Bus bars 51 and 52 also act as heat sinks for dissipating heat from chip 70.

FIG. 5 shows a second structure similar to that shown in FIG.
An example ZBJ head 20 is shown. The head 200 has a ZBJ chip 100 including nozzle arrays 102, 103, 104 and 105. The chip 100 has an ink channel 101, and each ink channel has an ink tank 21 of each color in a channel bulge 210.
1, 212, 213 and 214 are in communication.

The channel bulging portion 210 has another shape having a higher capacity than that shown in FIG. 4 for the chip 100 having the same size. Also, power bus bar 2
Tab connecting portion 20 for connecting 01 and 202 to the chip 100
3 and 204 are shown. The membrane filter 205 is also provided as described above.

In order to print an A4 size page, the head 200 is approximately 220 mm long and 15 mm wide.
A size of 9 mm depth is required. Many forms of ZBJ heads are possible using the above described basic arrangement. The actual size and number of nozzles per chip are simply determined by the performance requirements when applying the printer.

The physical structure of the ZBJ chip 100 will be described in detail below. The ZBJ chip 100 has, for example, four nozzle arrays 102 to 105 as shown in FIG.
This is achieved by four rows of nozzle paths (vias) 110 (see FIG. 6).
(A) to (D)). The nozzle path 110 is formed through the substrate 130 of the chip 100 by etching. Substrate 130 may typically be about 500 μm deep and 4 mm wide and 220 mm long depending on the required application. 6A to 6D show the substrate 13
Shown is the etching of the nozzle passage 110 through 0. ZB
In order for the J tip 100 to be able to eject 3 pl of liquid, the diameter of each nozzle 110 is approximately 20.
Requires μm. In one of the possible manufacturing methods,
In the range shown in FIG. 5, a four step process is used starting with a 500 .mu.m deep substrate 300 having a top glass (SiO2) layer 142 enclosing a heater 120. First, in the step shown in FIG. 6A, plasma etching is performed on a circular hole surrounded by a straight wall of 20 μm which penetrates the glass overcoat layer 142 and enters the substrate 130 at least 10 μm. This forms the nozzle tip portion 111.

In the next step, as shown in FIG. 6B, a large channel (approximately 100 μm) is formed from the back side of the chip 100.
Etching with a width of 300 μm). This forms a nozzle channel 114 that supplies the ink flow to the nozzle 110. In the next step, as shown in FIG. 6C, the channel 114 formed in FIG. 6B is formed.
Print the nozzle barrel pattern on the bottom of the. The nozzle barrel 113 has a depth of about 40 μm, and the tip 100
Plasma etching is performed within 10 μm in front of the. Since isotropic plasma etching is relatively non-selective, this method cannot be used to etch the entire volume without the heater penetrating and damaging the heater 120.

For this purpose, isotropic etching is used on the fully exposed silicon to a depth of 10 μm from the front of the chip 100, as shown in FIG. 6 (D). Due to the action of this process, the nozzle 110 is expanded and the heater 120 is expanded.
The lower side of the SiO ₂ layer 142 including is removed (undercut). In this process, the nozzle cavity (cavity) 11
2 is formed. This step also ensures that the nozzle tip 111 is connected to the barrel 113 without the risk of damaging the heater 120 with plasma etching. Those skilled in the art will appreciate that the above dimensions are only approximations and are presented only in general terms. However, the front-to-back etching should be better than 10 μm aligned and the control of the etching depth should also be better than 10 μm. In this way, the tip 111, the cavity 112, and the barrel 113
A complete nozzle channel 110 is formed that includes a channel 114 and a channel 114.

Nozzle tip 111, thermal chamber (thermal ch
It will be appreciated that the structure of the nozzle cavity 112, nozzle barrel 113 and nozzle channel 114, which act as an amber), form a path for ink flow through the substrate 100 for ejection.

For example, a conventional integrated bubble jet head manufactured by Canon Inc. uses hafnium boride (HfB ₂ ) as the heater element 120. Current Canon BJ10 (model number) printers have heater parameters with a liquid size of 65 pl selected. what if,
When the amount of liquid to be discharged is set to 3 pl, the size of the heater structure needs to be reset because it is substantially small.
A serpentine shape as shown in FIG. 7 can be used to ensure that the high temperature is reached, while maintaining the heater resistance and minimizing the overall size. Furthermore, as shown in FIG. 7, the heater 120 is composed of two heating elements in the form of a main heater 121 and a redundant heater 122, which surrounds the nozzle tip 111 and surrounds it. It is provided. The redundant heater 122 is provided to increase the fault tolerance of the ZBJ chip 100, thereby increasing the yield in the manufacturing process. The heater 120 has a BJ structure in which only one of the channel walls is formed due to the two-part structure.
In contrast to the prior art, where there is a heater on the chip.

FIG. 8 shows a fracture surface of the nozzle passage 110 shown in FIGS. 6 (A) to 7. In particular, the relative dimensions of the heater 120 and the nozzle tip 111 can be evaluated.

FIG. 9 shows a cross section taken along the line AA'-BB 'of FIG. 7 of the entire completed one nozzle heating chamber. The lower layer 130 is generally a silicon wafer having a thickness of about 200 μm (this wafer is thinned from a wafer having a thickness of 500 μm by back etching after high temperature treatment). In addition to having an ink passage and a heat transfer path for waste heat, the lower layer 130 also operates as a semiconductor substrate for drive electronics connected to the heater 120.

The heat insulating layer 132 is a 0.5 μm thick layer of thermally grown SiO ₂ . Layer 132 has several functions, including insulating the heater 120 from the upper passivation layer 144, acting as a mechanical cushion for bubble burst force heater 120, and MOS. It also includes operating as an integrated part of a drive circuit (discussed below).

In order to achieve the best heat transfer from the heater 120 to the ink 106, the heat insulating layer 1 can be used without impairing reliability.
32 is preferably manufactured as thin as possible. The heat insulating layer 132 is made of thermally grown Si instead of CVD SiO _2.
Since it is O ₂ , it has no pinholes. Therefore, the heat insulating layer 132 can be made thinner than the corresponding layer in the conventional bubble jet head using CVD SiO ₂ .

The heater 120 is a 0.05 μm thick layer of hafnium boride or other boride compound of a Group IIIA to VIA metal. This provides a high electrical resistance element for converting the electrical drive pulses into heating pulses. H
The very high fB ₂ melting point (3100 ° C.) means that it has a substantial margin at the actual heater temperature.
Electrical connection to the heater 120 is made by a heater connecting portion 123 made of aluminum having a thickness of 0.5 μm. It is formed as part of the first metal layer 134. The first metal layer 134 is an aluminum layer having a thickness of 0.5 μm. The first metal layer 134 is the heater 120.
It is formed at the same time as the connecting portion 123 connected to. This layer connects between the heater 120 and the drive electronics (described below),
Similarly, the inside of the drive circuit is also connected. It should be noted that for the color ZBJ heads described herein, there is a large number of nozzles in a small area, as high density, high quality linewidth interconnections are required. For this reason, interconnectivity of about 2 μm is required.

The intermediate heat insulating layer 136 is a CVD film having a thickness of about 1 μm.
SiO ₂ or PECVDSiO ₂ (PE is photon enhan
ced) provided as a layer. The thickness of layer 136 is ZBJ
It is important for the operation of the chip 100. because,
This thickness is associated with the heater 120 and the heat shunt.
t) 140, which provides a thermal offset between the ink 106 and most of the heat rather than the underlayer 130.
This is because it is surely transmitted to. The intermediate insulating layer 136 also
Electrical insulation is provided between the first metal layer 136 and the second metal layer 138. However, there is no limit to the thickness in this role.

A second metal layer 138 is provided to form a second layer for electrical connection similar to the heat shunt 140. Interconnection density of the above high speed ZBJ head (with 250 nozzles per 1 mm straight line)
Is high, two metal levels are required using the 2 μm design rule. Other head designs may have only one metal layer. If one metal layer is used, another configuration for the heat shunt 140 is required. This is because this metal layer is formed on top of the face 136 of the intermediate oxide.

The heat shunt 140 is formed from an aluminum disk approximately 0.5 μm thick. This shunt 140 is thermally connected to the lower layer 130 through the vias 410 in the insulating layer 132 and the intermediate layer 136, but serves no role for electrical purposes. The purpose of the heat shunt 140 is to dissipate waste heat from the heater 120 at a controlled rate in the lower layer 130.
To tie to. In order to keep the temperature of the ink 106 low during non-operation, the waste heat must be substantially moved during the period between the heater driving pulses.

The heat shunt 140 is designed to be formed simultaneously with the second metal layer 138. This is possible if the thickness of the heat shunt 140 matches the thickness of the second metal layer 138. The required thickness is determined by the nature of the thermal connection between heat shunt 140 and heater 120. The actual amount of heat connected is best determined by an accurate computer model for a particular geometrically defined nozzle. Another purpose of the heat shunt 140 is to prevent the thick CVD glass overcoat 142 from being heated.
This slows the diffusion of the CVD carrier gas through this thick layer 140 and thus delays the formation of bubbles that destroy the heater 120.

The overcoat layer 142 is CVD or P
A thick layer of ECVD glass that has three functions.
First, to provide nozzles for ejecting ink,
Second, it provides mechanical strength to resist the impact of the bursting or collapse of vapor bubbles, and third, it provides protection against the external environment. ZBJ chip 110
Must be exposed to the atmosphere for the printing process to operate, so its surface therefore needs to provide an increased level of protection over hermetically sealed devices. The thickness of the overcoat member 142 is about 4μ
It can be m, but this thickness can be substantially thicker corresponding to a suitable nozzle length.

The passivation layer 144 is made of a 0.5 μm thick layer of tantalum or other material and is used in the chip 1
It is adapted to the entire structure of 00 and covers it, providing chemical and mechanical protection.

Finally, the ink 106 in FIG. 9 not only has the obvious function of forming a printing mechanism, but also serves to transfer waste heat. 3 pl (picoliter) of ink liquid is approximately 1 for every elevated Celsius
Remove 3 nJ of heat.

FIG. 10 shows another heater configuration. here,
The heater 440 includes a main heater 441 and a redundant heater 443, each of which has an annular shape surrounding a nozzle 445 which ejects an ink droplet 446. Heater 44
1 and 443 are made from deposited HfB2 and are combined by overlapping aluminum connections 442 and 443, respectively. The heater 440 having this shape surrounds the thermal action cavity 447 existing in the lower portion, and therefore, the annular ink bubbles (FIG. 21A to FIG.
(D) and FIG. 22 (A) to (D)) can be generated. Therefore, the bubbles generate almost equal pressure over the entire circumference of the ink liquid 446. The main heater 441 and the redundant heater 443 are equal in shape and placement, so they have equal drop ejection characteristics. Further, the eccentricity of the heaters 441 and 443 with respect to the nozzle 445 is slight, so that even if the main heater 441 is damaged and the redundant heater 443 is used, the liquid ejection angle does not change significantly.

FIG. 9 has a cylindrical hole and a thin tip, and a thermal chamber on the lower side.
Although r), that is, a nozzle that forms a hole is shown, various nozzle shapes can be used. Some of them are shown in FIGS. 11 to 20.

In FIG. 11, the thermal action chamber 115 surrounding the nozzle tip portion 111 is formed in a cylindrical shape, and the heater 120 is deposited on the wall surface of the cylindrical shape. This arrangement has some disadvantages, including: (1) The heater film must be vertically deposited on the wall surface of the cylinder. Also, although this must be done by chemical vapor deposition, it is very difficult to make the heater 120 in the desired size and shape. (2) It is difficult to form a redundant heater to allow damage (described later). (3) The ink 106 and the vapor to be ejected are transferred to the tip 11
The heater 120 must be embedded inside the surface of the cylinder only to eject one. (4) ink is separated from the heater 120 by CVD SiO ₂ was used instead of crystalline SiO ₂ having high thermal conductivity.

In FIG. 12, the heat action chamber 115 has a conical shape. As a result, the resistance value of the heater 120 to be etched can be increased. This configuration has the following difficulties. (1) If the angle of the cone is too large (the cone is flat),
The ink 106 is not filled in the nozzle 110 due to the capillary phenomenon. (2) When the angle of the cone is too small (the cone is sharp), the heater 120 has the same shape as when the heating chamber has a cylindrical shape.
Are more difficult to fabricate. (3) The nozzle barrel (nozzle cylinder) 123 becomes very narrow, and the ink refill time increases.

FIG. 13 shows a chamber having a substantially hemispherical shape.
A frustoconical portion is provided facing the substantially hemispherical chamber, and a heater 120 is formed in that portion.

14 to 19 each show six preferred nozzle configurations. With these configurations, a single structure, 1
A small liquid size of 3 pl that enables a dot resolution of 600 dpi, a heater structure with a fault tolerance, and a nozzle that can be used in a multi-color printing device can be provided with a certain interval on either surface of the substrate.

FIG. 14 shows a heating chamber 1 having a substantially hemispherical shape.
Fifteen is shown. The heat chamber 115 is formed by hollowing out the lower side of silicon by isotropic plasma etching before the nozzle barrel 113 is formed by reactive ion etching (RIE). The feature of this configuration is that the bubble 116 is generated in the opposite direction to the ejection direction of the ink droplet 108. The heat shunt 140 guides heat from the nozzle area to the substrate 130, which reduces the time required to sufficiently cool the thermal chamber 115 prior to the ejection of the next ink drop 108.

This structure has the advantage that the heater 120 has a planar structure, which makes it possible to accurately control the shape and size of the heater. Further, the thermal coupling between the heater 120 and the ink 106 is important. Because the heater 120 is made of the ink 10 with the SiO ₂ layer 132 having a higher thermal conductivity than the CVD glass.
This is because it is separated from 6. Also, this layer is C
Compared with the case where a similar layer is formed by VD glass,
It can be made thinner without increasing the tendency for pinholes to occur. This nozzle configuration depends on the inclination of the portion of the barrel 113 entering the heat action chamber 115 and the contact angle between the passivation layer 144 (see FIG. 9) and the ink to automatically fill the ink by the capillary action. It is possible.

The various disadvantages of this structure reside in the above-mentioned adverse effect that the bubble generation is performed in the direction opposite to the ink ejection direction, and the efficiency of bubble generation ejection is reduced. It also requires a thick coating of CVD glass to form the nozzle area. In addition, wasteful heat must be dissipated through the long path of the silicon substrate 130, which extends over approximately 600 μm. This puts some limits on nozzle density and / or firing frequency. Another disadvantage of this configuration is that if the angle at which the barrel 113 enters the heat-acting chamber 115 is not precisely controlled, the nozzle 1 may be operated by capillary action.
It concerns potential difficulties in filling 10 with ink 106.

FIG. 15 shows a configuration similar to that of FIG. 14 except for the following layout configuration. That is, in FIG. 15, the direction of the flow of ink 106 through the chip 100 is opposite to the configuration shown in FIG. 14, providing a reverse nozzle configuration in which bubbles are generated in the same direction as the ink droplet ejection direction. As shown in FIG. 15, the ink 106
Enter the nozzle passage through the opening 484 and a meniscus 107 is formed at the nozzle tip 486 located at the boundary between the barrel 487 and the channel 489. Due to the action of the bubble 116, the ink drop 1 passes through the channel 489.
08 is ejected onto a medium such as paper 220.

The reverse nozzle configuration 485 differs from each of the previous configurations in one important respect. The configuration described above (eg, shown in FIG. 14) uses a heat shunt 140, which directs heat from the heater 120 to the substrate 130.

However, in the structure shown in FIG. 15, the ink storage chamber for the ink 106 exists below and adjacent to the heater 120. Therefore, in this configuration, the thermal diffusion path (diff
user) 491 is used to increase the area of heat transfer from the heater 120 through the coating layer 142 to the ink reservoir of the ink 106. In this configuration, the heat conduction path is considerably shorter than in the case of FIG. 14, so that a larger heat diffusion is achieved. Also, an ink supply pump (not shown)
Can be used to recirculate the ink 106 through the heat exchanger to further enhance heat diffusion.

The construction shown above has the advantages of a planar structure, good thermal coupling and thermal diffusion. Further, since the method of bubble generation is in the same direction as ink ejection, kinetic energy loss during ejection is reduced. The disadvantage of this configuration is that the ink cannot be self-injected into the nozzle and must first be pressurized to inject the ink. Once the ink is injected, the ink droplets 108 are ejected and then the ink is drawn into the thermal action chamber 488 due to the contraction of the bubbles. In addition, the cantilever-shaped portion that supports the heater 120 must have a sufficient thickness to withstand the impact when the bubbles 116 disappear.

Referring now to FIG. 16, there is shown a nozzle configuration having a heater 493 that is trench implanted. In this example, the nozzle cavity 112 is formed in the shape of a right cylinder, which is optionally etched by the nozzle barrel 113 and its formation.
etched) communicating with the nozzle channel 114. An annular groove 492 is formed in the silicon substrate by etching, and SiO ₂
Layers are layered near the nozzle cavity 112. An annular heater 493 is formed on the inner surface of the groove 492. Heater 4
93 is a bubble 11 due to the vaporization of the ink.
6, which bubbles grow across the cavity 112 in a direction transverse to the ejection direction. Among the advantages of this configuration are good thermal coupling and self-injection structure. Further, one of the disadvantages is that the heat diffusion is poor. This is because most of the ink is 600μ from the bubble generation surface.
This is because the ink is not efficiently cooled due to the flow of the ink because it is separated by the substrate 130 of m. Further, since the nozzle length is determined by the extremely thick CVD glass layer forming the coating layer 142, it is difficult to determine an appropriate nozzle length. Furthermore, since the bubbles are generated in the transverse direction, it is impossible to obtain the optimal kinematic relationship for ejection. In addition, since the length of the heater 493 is limited to the circumference of the nozzle or to the half of the circumference when the fault tolerance structure is used, it is difficult to increase the resistance value of the heater 493. is there.

FIG. 17 shows the above-mentioned reverse nozzle structure as shown in FIG.
The case where the annular groove structure of is used is shown. Here, the annular groove 49
2 extends in the direction of the nozzle tip 486 along with the heater 493 formed diffused therein. The thermal diffuser 491 is also provided in the same manner as described above. The advantages of this configuration are found in thermal coupling, ease of thermal diffusion, self-filling and ease of manufacture. Disadvantages include that the bubble generation direction is transverse to the ink droplet ejection direction, and it is not easy to change the length of the heater 493 in various ways. Further, since the proportion of heat conducted to the silicon substrate 130 becomes high, the heat wasted may be increased.

FIG. 18 shows a structure using an elbow heater. In this example, the cylindrical nozzle passage has a nozzle tip 11
1 and the barrel 113. The SiO2 layer 494 obtained by thermal growth extends into the barrel 113 side. Furthermore, the heater 4 extending in an elbow shape
95 is deposited on the electrical contacts (not shown) formed on the upper surface of the layer 494 and the heater 495. Furthermore,
A coating layer 142 of CVD SiO2 is provided on the layer extending toward the contact, heater and nozzle barrel 113. The advantage of this configuration is that the self-filling and heater 49
5 is in thermal isolation to the substrate 130. In addition, disadvantages include poor thermal diffusion, difficulty in adjusting the nozzle length that requires changing the thickness of the coating layer 142, and a bubble generation direction that is transverse to the ejection direction. The heater 495 is separated from the ink 106 by a layer of amorphous CVD glass so that the heater does not have good thermal coupling with the ink, it is difficult to change the length of the heater, and manufacturing is complicated. (Described later), etc.

FIG. 19 shows an inverted structure nozzle using an elbow contact heater 495 formed in the same manner as the above structure. The advantages of this configuration can be found in the thermal diffusion through the ink reservoir, self-filling, and the heater 495 being thermally isolated from the substrate 130. In addition, as a disadvantage, the bubble generation direction is transverse to the ejection direction, the thermal coupling between the heater 495 and the ink 106 is not performed satisfactorily because of the amorphous CVD glass, and the heater. It can be mentioned that the length is limited.

FIG. 20 shows a nozzle structure similar to that shown in FIG. However, the relative size of the nozzle opening 484 with respect to the nozzle tip 486 is different from the configuration of FIG. 15, which may improve capillary action for ink filling at the nozzle and formation of the meniscus 107. That is, one of the disadvantages of the configuration shown in FIG. 15 is the nozzle opening 484 and the nozzle tip 486.
And have the same diameter. Nozzle tip 48
The diameter of 6 will vary depending on a number of design criteria such as required liquid size.

As shown in FIG. 20, in order to fill the nozzle with ink and form the meniscus 107, the ink 10
6 must flow through opening 484 and stop at flow tip 486. When the opening and the tip have the same size, generally, a meniscus is formed in the opening 484 or ink drips from the tip 486 depending on the filling pressure of the ink. Neither of these situations is desirable. A particularly desirable configuration is that the opening 484 has a diameter sufficient to allow ink filling of the nozzle and the tip 4
86 is different in that it has a smaller diameter that is used to form the meniscus 107. Therefore, the nozzle is greater than the "bubble pressure" at the opening 484 and the tip 48
The ink is filled using a pressure less than the "bubble pressure" in 6. The configuration shown in FIG. 20 shows a preferred structure for the opening and tip, where the diameter of opening 484 is about 50% larger than that of tip 486. Further, according to this configuration, it is possible to accurately control the size of the liquid while maintaining a high refill speed.

The operation of the ZBJ chip 100 is shown in FIG.
1 (A) to (D) and FIG. 22 (A) to (D), the use of the novel liquid ejecting mechanism is different from that of the conventional bubble jet head.

FIG. 21A shows one nozzle 110 of the ZBJ print head 100 in its rest state with the heater 120 off. The ink 106 in the nozzle 110 forms a meniscus 107. Figure 21
In (B), the heater 120 is driven to heat the substrate 130 and the thermal layer 132 around the heater 120, thereby heating the ink 106 in the nozzle 110. As a result, the ink 106 is partially evaporated to form micro bubbles 116. As shown in FIG. 21C, since the evaporated ink is heated, these micro bubbles expand and coalesce into large bubbles 116. In FIG. 21D, the pressure of the expanding bubble 116 pushes the ink 106 out of the nozzle tip 111 at high speed.

In FIG. 22A, the driving of the heater 120 is stopped, whereby the bubble 116 contracts and pulls the ink from the formed liquid 108. In FIG. 22B, the liquid 108 is separated from the ink 106 in the nozzle 110, and the contracting bubbles 116 pull the meniscus 107 toward the rear of the nozzle 110. FIG. 22 (C)
As shown in FIG.
Ink 10 is refilled with ink from the lower storage chamber. At this time, the ink is overfilled due to the refill ink speed. Finally, in FIG.
After the meniscus of No. 6 vibrates, it finally returns to the rest state shown at the beginning. The time during which this vibration decays is one factor that determines the maximum dot frequency.

When the heater 120 is driven as shown in FIG. 23, a part of the heat generated by the heater 120 is transferred to the ink 106 and the rest is transferred to the member around the nozzle, as indicated by the arrow.

FIG. 24 shows superheat of ink,
According to this, a thin layer of superheated ink 1
09 is formed adjacent to the passivation layer 144 in the nozzle cavity 112.

After the driving of the heater 120 is stopped, the excess heat must be quickly removed. Heater 120
Ink temperature is 100 ° C within 200 μsec
Beyond the above, since the ink 106 is substantially water, bubbles are formed without producing ink residue. If such a state does not occur, the heater 120 and the ink 106
Since there is a heat insulating layer between them and the ink, ink droplets cannot be ejected accurately.

Excess heat is removed via three separate paths. First, heat is removed via the ink. At this time, the temperature of the ink slightly rises. However,
Since the thermal conductivity of the ink is low, the amount of heat removed by this path is small.

The wall of the nozzle 110 is formed by the silicon of the substrate 130, and since this silicon has a high thermal conductivity, the heat diffusion through this wall takes place at a high rate. However, all parts of the bubble 116 are
It is not in contact with this side wall of 10.

Further, excess heat is removed via the heater 120. Thermal diffusion through the heater 120 is important because the ink vapor must not contact the heater 120 when the liquid is ejected. Since most of the material around the heater 120 is glass with low thermal conductivity, a heat shunt 140 is provided therein to shunt excess heat to the substrate 130.
Removal of excess heat is almost 200 μs without providing a heat shunt
It is not necessary to provide such a heat shunt 140 if it ends during ec. FIG. 25 shows the flow of heat when the bubbles 116 are cooled.

FIG. 26 also shows a path for removal of excess heat, where heat flows through the substrate 130 as the main heat conduit from the heater 120. A part of this heat returns to the ink 106 and is finally released together with the next liquid 108 when it is ejected. The rest of the heat is substrate 1
Aluminum heat sink (51, 52: FIG. 4)
See).

Ink Channels As shown in FIGS. 28 and 29, the ZBJ printhead 200 has, for example, ink channels 211, 212, 213 and 214 corresponding to different probe solutions. These channels 211-214 are
It is formed as an aluminum extruded body and performs filtering and sealing on the back surface of the ZBJ chip 100.

In some applications, the ink channels 211-214 shown in FIG. 28 may not be sufficient for proper ink flow. In such a case, FIG.
An extrudate geometry such as that shown in Figure 9 can be used to increase the amount of ink flow. As shown in FIG. 28, the channel pusher 210 and the ZBJ tip 1
00, an absolute membrane filter 205 having a pore diameter of 10 μm is provided, which can protect the head from ink contamination. If the membrane filter 205 has compressibility, this filter also functions as a gasket and can prevent the four color inks from mixing with different colors. Both ends of the head assembly 200 are preferably sealed to prevent gas ingress. With respect to the above configuration, the chip 100 and the channel pusher 210 which are required to be maintained in manufacturing are required.
The alignment accuracy of is about ± 50 μm.

The ink jet head used for manufacturing the two-dimensional probe array as described above uses the structure of the ZBJ head described above as its prototype, and the liquid to be ejected is replaced with a water-based ink to form a probe solution. Can be designed. That is, since the ZBJ head has an ink inlet on one side in the Z direction and a nozzle on the other side of the integrally formed head chip, the direction of the ink flow path and the ejection direction is Is linear and never orthogonal. Therefore, even if the ZBJ head has a form having a large number of solution reservoirs and nozzles corresponding thereto, it is relatively easy to form the number of channels corresponding to the number of solution reservoirs and nozzles. As a result, the ZBJ head manufactured in this way is suitable for the present invention which needs to eject a large number of probe solutions.

FIG. 30 is a schematic view showing an example of an ink jet head in which nozzles are arranged in a two-dimensional array, which is used in the present invention. Such an inkjet head can be manufactured by appropriately selecting and applying the manufacturing method of the ZBJ head described above. As shown in FIG. 30, in this liquid ejecting apparatus, nozzles and corresponding solution reservoirs are integrally incorporated in a two-dimensional array. In some cases, inkjet
The head may be configured by a one-dimensional array, or it is not an integral type and is, for example, one in which the top and bottom are stuck together, or a plurality of one-dimensional inkjet heads are arranged and nozzles are arranged in one two-dimensional array. It may be an inkjet head.

The amount of the probe solution discharged from one nozzle at a time takes various factors such as the viscosity of the probe solution, the affinity of the probe solution and the solid phase substrate, and the reactivity of the probe and the solid phase substrate into consideration. The above is appropriately selected according to the dot size and shape of the array to be formed. An aqueous solvent is generally used as the probe solution, and in the method of the present invention, the liquid of the probe solution ejected from each nozzle of the inkjet head is generally 0.1 picoliter. To 100 picoliters, and the nozzle diameter and the like are preferably designed and adjusted according to the liquid volume.

Further, although each liquid of the probe solution discharged onto the solid phase substrate for forming the probe array is originally circular in shape, a plurality of fine circular dots are connected to each other. A rectangular coating surface can also be used. The area occupied by the array units (dots) to which this probe solution is applied is 0.01 (for example, 0.1 μm × 0.1 μm)
μm ² to 40000 (eg 200 μm × 200 μ
m) μm ² is common, which depends on the size of the array itself and the density of the array matrix.

The size and capacity of the solution reservoir are also appropriately selected depending on the amount of probe solution discharged from the nozzle at one time and the number of arrays to be prepared.
I can't make a general decision. In addition, inkjet
When the head is processed into a solid phase substrate such as a silicon substrate to form an integrated type, the nozzle diameter and the amount of the probe solution discharged from the nozzle at a time are naturally within a certain range.
In some cases, the size formed on the surface of the substrate facing the opening of the nozzle is sufficient, as in the example of FIG. On the other hand, when the amount of probe solution ejected from the nozzle at one time is relatively large and the number of arrays required to be manufactured is large, the probe solution is added to the solution reservoir to produce the total number of arrays. A method to cover the required amount of solution can be considered. Apart from that, the structure may be such that it is attached to a solution reservoir arranged on one surface of the substrate and further has a solution storage portion for increasing the volume. At that time, the addition of the probe solution is carried out via the solution storage part for increasing the volume, and the shape of the solution storage part itself can be set so that the probe solution can be easily supplied if necessary.

If necessary, the solution storage section may be provided with a nozzle,
The structure may be integrally formed in an array shape corresponding to the array and overlapped with the liquid ejection device. However,
When the solution storage part has a desired large capacity, a nozzle array,
If the space for arranging such a large capacity cannot be secured with the density as it is due to the density of the solution reservoir, etc.,
It is also possible to adopt a configuration in which a larger-capacity solution storage portion and a solution reservoir are connected by a flow path.

Regarding the form of the liquid ejecting apparatus, it is desirable that the liquid ejecting apparatus is integrally formed in a two-dimensional array. However, a plurality of one-dimensional array liquid ejecting apparatuses are combined to form an integrated body to form a two-dimensional array. You may use the liquid discharge apparatus made into the shape.

Regarding the ink jet system to be used, a detailed explanation has been given here by exemplifying a structure adopting the thermal jet system. However, depending on the density of the array, the liquid to be ejected may also be used. Depending on the amount of the liquid, it is possible to use a piezo jet method to similarly form a two-dimensional array.

For each probe used in this array, a pair of solution reservoirs and nozzles are made to correspond and arranged in an array. Therefore, the number of solution reservoirs and nozzles forming a pair is not particularly limited, and is selected according to the probe type of the array required. In addition,
Although the dot diameter, the number of dots, the coating density, or the array shape is naturally limited by the manufacturing method of the liquid ejection device, the total number of paired solution reservoirs and nozzles that make up the device is basically Is determined by the probe species, and in some cases may be 100,000 or more.

In the method of the present invention, generally, the probes arranged in a two-dimensional array on the substrate are of the same kind in a broad sense. That is, in the method of the present invention, the type of each probe itself is not particularly important as long as each probe can be ejected as a solution from the liquid ejecting apparatus. On the other hand, the method of the present invention is applied to those that can be fixed on the substrate after being discharged and applied as a solution on the substrate. As a probe that meets this requirement, for example,
DNA, RNA, cDNA (complementary DN
A), PNA, oligonucleotides, polynucleotides, other nucleic acids, oligopeptides, polypeptides, proteins, enzymes, substrates for enzymes, antibodies, epitopes for antibodies, antigens, hormones, hormone receptors,
Ligands, ligand-receptors, oligosaccharides and polysaccharides can be mentioned as examples.

It is desirable that these probes have a structure capable of binding to the solid phase substrate, the probe solution is discharged and applied, and then the probe solution is used to bind to the solid phase substrate. The structure capable of binding to this solid-phase substrate is, for example, an amino group, a mercapto group, a carboxyl group, a hydroxyl group, an acid halide (-COX), a halide, an aziridine, a maleimide, a succinimide, an isothiocyanate, a sulfonyl chloride (-SO ₂ C
l), aldehyde (-CHO), hydrazine, iodoacetamide and the like can be formed by performing a treatment for introducing an organic functional group into the probe molecule in advance. In that case, on the other hand, it is necessary to previously perform, on the surface of the substrate, a structure for reacting with the above various organic functional groups to form a covalent bond and a treatment for introducing the organic functional groups.

2) Example of Embodiment Using Liquid Ejecting Device Next, the nozzles used for ejecting the liquid of the probe solution onto the solid phase substrate to form the dot-shaped application region of the probe solution are arranged in a two-dimensional array. A liquid ejecting apparatus using the thermal jet method will be described.

(Structure of Liquid Ejecting Device for Ejecting Probe Solution) (Embodiment 1) FIG. 31 is a schematic view of an example of a liquid ejecting device using a semiconductor chip by a thermal jet method according to the present invention as a substrate. Note that FIG. 31 is a schematic view of the surface on the front side of the semiconductor chip in which the nozzles, flow paths, and the like that compose each liquid ejection portion incorporated in the liquid ejection device are formed.
As will be described later in detail, in this example, one liquid ejection device is configured by using a plurality of the chips. It should be noted that the present invention may configure a liquid ejection device by using such a chip alone.

Most of the functions of the liquid ejecting apparatus depend on the structure of the chip. In addition to the semiconductor chip made of Si or the like, a plate-shaped chip made of glass can be preferably used as the substrate.

FIG. 32 is an enlarged view of a portion near the nozzle in FIG. 31, that is, an enlarged view of a portion surrounded by a circle in FIG. 31 and 32, 1 is a silicon substrate as a semiconductor substrate, 2 is a heater element as a liquid ejection energy generating element made of TaN, TaSiN, TaAl or the like, 3 is a first wiring made of aluminum or the like, 4 is aluminum or the like A second wiring consisting of 5 is a pad for making electrical contact between the liquid ejection device and the outside, 6 is a nozzle for ejecting liquid from its opening, 7 is a flow path, and 8 is from the back surface of the substrate to the front surface of the substrate. A supply port for supplying a liquid. Although the supply port will be described later, the supply port is formed by anisotropic etching of silicon. At that time, the size of the supply port is widened as indicated by 9 (dotted line portion) on the back surface of the substrate 1. The supply port also serves as a liquid reservoir.

The direction indicated by the white arrow in FIG. 31, which will be described in detail later, is the direction in which the liquid ejecting apparatus moves, and is hereinafter referred to as the main scanning direction.

As shown in FIG. 31, the chip has a lateral nozzle interval of 1.27 mm (corresponding to 20 dpi).
Nozzle group consisting of 8 nozzles arranged in 5
The columns are arranged offset from each other at intervals of 254 μm (corresponding to 100 dpi).

The "offset" means an arrangement in which there is a deviation between the nozzle rows adjacent to the position of each nozzle constituting the nozzle row. For example, taking the first nozzle from the left side of each nozzle row extending in the horizontal direction in FIG. 31 as an example, the first nozzle of the second row counting from above is the first nozzle of the first row above it. It is arranged on the left side of the first nozzle with a gap of a predetermined distance. Furthermore,
The first nozzles in each of the third to fifth rows are also sequentially arranged on the left side with a similar gap between them. Since the interval between the adjacent nozzles in the same row is constant, the second to eighth nozzles in each row are also arranged with a predetermined deviation. According to the offset arrangement in the example of FIG. 31, there are no overlapping nozzles in the main scanning direction, and in the direction corresponding to the lateral direction of FIG. 31 of the spot formed on the solid phase substrate by the probe solution discharged from each nozzle row. It is possible to effectively increase the density (see, for example, the spot in the first column indicated by ● in the vertical direction in FIG. 42).

The distance between adjacent nozzle groups (vertical distance in FIG. 31) is 1.27 mm (equivalent to 20 dpi).
Is set to.

The eight heaters included in the one set of nozzle groups are connected by a set of first wiring and second wiring. The length of the tip in the long side direction is about 12 mm,
The length in the short side direction is about 7 mm. The number of nozzles in one chip is 40.

As shown in FIG. 31, the heater element has its both ends connected to the first and second wirings.

FIG. 33 is a sectional view taken along the line AA 'in FIG. 34 is a sectional view taken along line BB ′ of FIG. FIG. 35 shows a schematic view of the back surface of the semiconductor chip (corresponding to the back surface of the semiconductor chip shown in FIG. 31).

In FIGS. 33 and 34, the same reference numerals as those used in FIGS. 31 and 32 denote the same items.
Further, 10 is an insulating film, 11 is a protective film, 12 is a nozzle material,
Reference numeral 13 is a cavitation resistant film made of Ta or the like.

The insulating film 10 may be any film such as a thermal oxide film formed by thermally oxidizing a silicon substrate, an oxide film or a nitride film formed by CVD. The protective film 11 has a function of maintaining insulation between the wirings to be insulated, and also has a function of protecting the heater element and the wiring from liquid, and is provided at least in a portion required for such a purpose. . This protective film is an oxide film formed by CVD,
Any film such as a nitride film may be used. The nozzle material 12 forming the nozzle 6 and the flow path 7 can be formed of glass or the like,
The formation of the flow path and the like on the substrate is performed even if a nozzle material having a nozzle and a flow path is attached to the semiconductor substrate in advance.
It may be directly formed on a semiconductor substrate by using a semiconductor process using a photolithography technique.

In particular, when the semiconductor chip needs to be manufactured in a larger area, it is difficult to manufacture the chip by the method of attaching the nozzle material having the nozzle and the flow channel to the semiconductor substrate. Therefore, the photolithography technique is used. A manufacturing method using is desired.

The supply port 8 is formed by anisotropic etching of silicon using a TMAH solution, and is opened at an angle of 54.7 degrees with respect to the substrate surface, as shown in FIG. The shape of the supply port is a truncated pyramid as shown in FIG. If the width of the supply port on the front surface of the substrate is set to 100 μm as shown in FIG. 33 and the thickness of 1 of the silicon substrate is 625 μm, the width on the back surface of the supply port is about 1 μm.
mm.

The position of the supply port 8 is determined by the photolithography technique, for example, with respect to the heater element 2. When the nozzle is also formed by photolithography as described above, the relative positional relationship between the nozzle and the liquid reservoir can be defined very accurately. For example, the direction of liquid supply from the supply port and the liquid supply from the nozzle can be defined. The length of the flow path 7 in the liquid flow direction can be designed to be a very short distance by making the discharge directions the same. As a result, it is possible to suppress the problem that bubbles are accumulated in the flow path.

In the conventional inkjet head for printing, the main purpose of the supply port is to guide the ink from the ink tank provided on the back surface of the substrate to the heater section.
As described above, in the probe array manufacturing liquid ejecting apparatus, since the total amount of ejected liquid is small, the supply port can be used as a liquid reservoir integrally formed on the substrate.

At the supply port of the above size, the volume is about 0.23.
Since the ejection amount is 24 pl in this example, this volume corresponds to an amount capable of producing about 9600 probe arrays.

When the chip is viewed from the back side, the supply port is shown in FIG.
The shape shown in FIG. As shown in FIG. 34, the liquid is guided from the back surface of the substrate to the front surface of the substrate from the supply port 8 and then to the nozzle 6 through the flow path 7.

When a voltage is applied across the heater element,
The liquid in the vicinity of the heater is overheated to cause film foaming (film boiling), and the liquid is discharged as shown in FIG. The amount of liquid ejected from the nozzle was 24 pl.

In order to stably eject the liquid, it is essential to stably cause the film foaming. It is desirable to apply a voltage pulse of 0.1 to 5 μs to the heater in order to cause stable film foaming.

Further, in order to precisely control the amount of each probe included in the probe array, the amount of liquid ejected from the nozzle is stable.
No. 0, JP-A-4-10941 and JP-A-4-10941.
A method in which bubbles generated by being heated by a heater communicate with the outside air as described in Japanese Patent No. 10942 is preferable.

FIG. 38 shows a schematic diagram of a liquid ejection apparatus according to the present invention. In FIG. 38, 21 is a liquid ejection device, 22 is a semiconductor chip described in FIGS. 31 to 37, and 23 is a nozzle. In FIG. 38, the tip 22 shows only the nozzle 23 in order to clarify the point. In this example, a case where a liquid ejection device is manufactured using 25 semiconductor chips 22 will be described. The semiconductor chips 22 are arranged in 5 rows and 5 columns, and each chip has 40 nozzles.
It can be seen that one liquid ejection device has 1000 nozzles.

FIG. 39 shows the structure of the liquid ejecting apparatus in more detail. The liquid ejecting apparatus is constituted by attaching a chip 22 to a substrate 24, which is made of alumina, resin or the like and has a window frame-shaped hole, as shown at 25 in FIG. The pads of each chip are electrically connected to the outside of the liquid ejection device by a flexible wiring board (not shown). The probe solution is injected into the supply port 8.

Embodiment 2 FIG. 40 shows another example of the liquid ejection device. In FIG. 40, 26 is the supply port 8 for the chip 22.
This is a substrate made of alumina, resin, etc., with holes 27 formed at positions corresponding to. In this example, the hole 27 formed in the substrate 26 also functions as the second reservoir, and the volume of the reservoir is the sum of the volume of the supply port and the volume of the second reservoir. The volume of the second reservoir is the substrate 26
It can be freely changed by changing the thickness of the hole and the size of the hole 27.

Further, instead of making a hole, a tube may be connected to the hole to supply the liquid.

(Structure of probe array manufacturing apparatus) FIG.
FIG. 1 shows a schematic diagram of the structure of a probe array manufacturing apparatus using a liquid ejection apparatus. In FIG. 41, 22 is a liquid ejecting apparatus, 31 is a shaft for guiding the movement of the liquid ejecting apparatus section substantially in parallel, 32 is a stage to which the probe array is fixed, and 33 is a glass substrate to be the probe array.
The liquid ejection device moves in the X direction in FIG. 41, and the stage moves to the Y direction.
By moving the direction, the liquid ejecting apparatus can move in a two-dimensional manner relative to the stage.

Although FIG. 41 shows the structure in which a plurality of glass substrates to be probe arrays are fixed and the probes are applied, the probe arrays are manufactured on one glass substrate to be a large probe array. After that, the glass substrate may be cut to obtain a probe array.

(Probe Solution Coating Method Using Liquid Ejection Device of Thermal Jet Method) Next, a probe array manufacturing method will be described.

The following is an example of the arrangement and control of the nozzle group in the case of using the liquid ejection device in which the nozzle openings are arranged in a two-dimensional array using a plurality of chips.

A) The openings of each nozzle formed by combining a plurality of chips are arranged in a two-dimensional array, the nozzles in each chip are arranged in a two-dimensional array of m rows and n columns, and n belonging to the same column The heater element corresponding to each nozzle is
A configuration that is connected to a common first and second wiring.

B) The openings of each nozzle formed by combining a plurality of chips are arranged in a two-dimensional array, the nozzles in each chip are arranged in a two-dimensional array of m rows and n columns, and m belonging to the same column. The heater element corresponding to each nozzle is
A configuration in which a common heater group connected to the first and second wirings is configured, and each heater group is sequentially driven at a desired timing.

C) The openings of the nozzles, which are formed by combining a plurality of chips, are arranged in a two-dimensional array, and the nozzles in each chip are arranged in a two-dimensional array of m rows and n columns.
The nozzle group in the X-th column (1 ≦ X ≦ n) composed of individual nozzles is arranged with an offset from the adjacent nozzle group.

D) The openings of the nozzles formed by combining a plurality of chips are arranged in a two-dimensional array, and the nozzles in each chip are arranged in a two-dimensional array of m rows and n columns.
The number of adjacent nozzles is Y.
The nozzle group of the X-th column (1 ≦ X ≦ n) arranged in is arranged with an offset from the adjacent nozzle group,
The offset distance is Y / n.

E) The openings of the nozzles, which are formed by combining a plurality of chips, are arranged in a two-dimensional array, and the nozzles in each chip are arranged in a two-dimensional array of m rows and n columns.
The nozzle group in the X-th row (1 ≦ X ≦ n) composed of a plurality of nozzles is arranged with a distance Z from the adjacent nozzle group, and after ejecting from the X-th nozzle group, liquid ejection is performed. A configuration in which ejection is performed from nozzle groups in adjacent columns at the timing when the device moves a distance Z.

Furthermore, by controlling the timing and pattern of liquid ejection from the nozzles in the liquid ejection apparatus of the present invention, the probe solution is deposited on the solid phase substrate at a density higher than the nozzle density of the liquid ejection apparatus used. Can be discharged.

An example of a method of manufacturing the probe array will be described below with reference to the drawings. FIG. 42 shows a schematic diagram for explaining a method for applying a probe solution using a liquid jet device of the thermal jet system. 41 to 4 in FIG.
2 is a semiconductor chip, and 33 is a probe array. In FIG. 42, the surface of the probe array is drawn so as to face up. Therefore, the arrangement of the nozzles of the chip and the main scanning direction are opposite to those shown in FIG. 38.

42 is a chip shown by "1" in FIG. 38, and 42 is a chip shown by "2" in FIG. As shown in FIG. 1, the structure of the chip is such that nozzle groups each composed of eight nozzles are arranged offset from the adjacent nozzle groups. Since the heaters in one nozzle group are connected to one set of common first and second aluminum wirings as described above, a voltage pulse can be applied between one set of pads connected to these wirings. By doing so, it is possible to cause the liquid to be ejected simultaneously in the nozzles that form the same nozzle group. That is,
Upon application of the voltage pulse, the ejection of liquid takes place simultaneously in the eight nozzles.

38 to 42, the numbers 1, 2, 3, 4, and 5 displayed adjacent to the tip are displayed to show the numbers of the nozzle groups, that is, A, B, and C,
D, E, F, G, and H are objects displayed to distinguish the individual nozzles that make up each nozzle group.

First, in FIG. 42, 1A, 1B, 1C, 1D, 1E, 1F, 1G,
Eight probes labeled 1H are formed. Next, the liquid ejection device ejects from the second nozzle group at the timing when the liquid ejection device moves 1.27 mm (corresponding to 20 dpi) in the main scanning direction, and 2A, 2B, 2C, 2D, 2E, 2F, in FIG.
Eight probes, indicated by 2G and 2H, are arranged in a row on the probe array. Then, by the same operation, the ejection is sequentially performed from the third to fifth nozzle groups,
In the probe array 33, 40 probes (indicated by black circles in FIG. 42) in the first row are arranged.

At this time, the distance between the centers of adjacent probes is 254 μm (corresponding to 100 dpi).

That is, 40 discharged from the first chip
The types of liquid form the first row of probes in the probe array. This state is schematically shown by 43 in FIG. (Only 5 probes of 1A to 5A out of 40 probes are shown.) Next, liquid is discharged from the second chip by the same operation, and the second row of the probe array (see FIG. 12).
The probe indicated by the middle white circle) was placed.

At this time, the center-to-center distance between the first row of probe groups manufactured from the first chip and the second row of probe groups manufactured from the second chip was 254 μm.
The drive timing was adjusted so that m (corresponding to 100 dpi) was obtained.

As in the case of the first chip, the 40 kinds of liquid ejected from the second chip form the second row of probe groups in the probe array. This state is shown in FIG.
It is shown schematically in 44. (Only 5 probes of 1C to 5C out of 40 probes are shown.) Further, liquid was ejected from the 3rd to 25th chips by the same operation to fabricate a probe array. (Probes surrounded by dotted lines in FIG. 42) As described above, a 100-dpi, 40-row, 25-column probe array can be produced from the liquid ejection apparatus shown in FIG.

It can be easily understood from the above description that a probe array requiring a larger number of probes can be manufactured with the same configuration.

The case where silicon is used as the chip which constitutes the liquid discharge device has been described above.
As described above, since the present invention has a simple structure including a heater and wiring connected to the heater, it is not necessary to use a silicon substrate, and a similar liquid ejecting apparatus using a cheaper substrate such as a glass substrate can be used. Can be made.

In the above example, the case where semiconductor chips having 40 nozzles are arranged in 5 rows and 5 columns to form a liquid ejecting apparatus has been described, but the number of nozzles and the arrangement of the arrangements are shown in the embodiment. The configuration is not limited to this, and can be freely selected.

In the case of using the semiconductor chip described in the above example to fabricate a liquid ejecting apparatus in a one-dimensional array such as one row and twenty-five columns, a probe array can be produced only by main scanning. The structure of the device can be simpler than that shown in FIG.

Further, although the method of manufacturing a liquid ejection apparatus using a plurality of chips has been described, the number of chips is increased because the chip structure is simple and a high yield can be obtained even if the chip size is increased. It is also possible to manufacture a chip having the nozzles of No. 1, and for example, it is also possible to manufacture a chip having 1000 nozzles.

When a probe array having 1000 probes is manufactured using this chip, the work of aligning a plurality of chips and adhering them to a member of the liquid ejection apparatus becomes unnecessary, and the structure is simpler. A liquid ejecting device can be manufactured.

Although the method of arranging the probes of the probe array at a density higher than the nozzle density of the liquid ejecting apparatus has been described above, the nozzle density of the liquid ejecting apparatus and the probe density of the probe array are the same. But it's okay. In this case, the arrangement of the nozzles is configured to have no offset as shown in FIG. Further, in this manufacturing method, it is not necessary to scan the liquid ejecting apparatus when manufacturing one probe array (another coating method of the probe solution using the liquid ejecting unit of the thermal jet method).

Next, another method for preparing the probe array will be described.

Up to now, the method of applying the probe solution when using the chip in which the nozzle groups adjacent to each other are arranged offset from each other has been described, but here, the nozzles are arranged on the orthogonal coordinates. The method of applying the probe solution in the case of being present will be described.

A method for producing a probe array having a density higher than the nozzle density by using the liquid ejecting apparatus having the structure shown in FIG. 44 will be described below.

Consider now that the nozzle density is 20 dpi, that is, the interval between adjacent nozzles is 1.27 mm. Use this tip to create a 100 dpi density probe array.

FIG. 44 is a diagram for explaining a method of manufacturing a probe array of 100 dpi with a chip of 20 dpi.

Since the probe pitch of the probe array is 100 dpi with respect to the nozzle pitch of 20 dpi, it is impossible to arrange the probes in one drawing operation.

Therefore, when a chip having a nozzle array of 8 rows and 5 columns as shown in FIG. 43 is used, the probe array is formed by repeating the drawing operation a total of 8 times. One drawing operation scans the liquid discharge device in the direction of the arrow (main scanning direction) in the figure, and when the nozzles arranged in five rows pass the predetermined position (corresponding to 100 dpi), drive the heater and move horizontally. Form the probe rows so that they are lined up at 100 dpi in the direction.

The probe indicated by ● represents a probe ejected in the first scan, and the probe in the first row (from the lower side) of the probe array is formed by the first scan.

Next, as shown in the figure, the head is shifted in the sub-scanning direction by a shift amount of 1.016 mm, and the drawing operation is performed in the same manner as the first scan. In this way, by repeating the drawing operation while shifting the position of the liquid ejecting head in the vertical direction by 5 probes by one scan, 8 vertical probes arranged at 100 dpi are completed.

Here, 8 rows with a nozzle spacing of 1.27 mm (20 dpi).
Tip with 5 rows of nozzles, probe spacing 0.254 mm
Although the method of manufacturing a probe array of 8 rows and 5 columns of (100 dpi) has been described, the number of nozzles and the arrangement of the arrangement are not limited to this, and a probe array with a desired number of probes can be obtained by the same operation. It can be made.

Further, when the probe arranging method as described above is carried out, it is necessary to set the discharge timing of the probe discharged from each nozzle to a desired timing. Therefore, the heaters corresponding to the respective nozzles are different from each other. It has a structure that can be controlled independently. Specifically, the wiring to be connected has a structure in which one end of the wiring is a common wiring and the other end of the wiring is independently drawn for each heater.

[0210]

EXAMPLES The present invention will be described in more detail with reference to the following examples. Although the examples shown here are examples of the best mode for carrying out the present invention, the present invention is not limited to these examples.

(Reference Example) A. Preparation of Solid-Phase Substrate After ultrasonically cleaning a 25.4 mm × 25.4 mm × 0.5 mm ^t fused quartz substrate in 1% ultrasonic cleaning detergent GP-III (Branson) for 20 minutes, it is then treated with tap water. Sonic cleaning and running water cleaning were appropriately performed. Next, 80 ° C. 1N Na
It was immersed in Cl for 20 minutes, washed with running water (tap water), ultrasonically cleaned with ultrapure water, and washed with running water (ultra pure water).

Purified by distillation under reduced pressure, the following formula (I):

[0213]

[Chemical 1]

Aminosilane coupling agent (KBM-
603: An aqueous solution containing Compound I (manufactured by Shin-Etsu Chemical Co., Ltd.) at a concentration of 1% was stirred at room temperature for 1 hour to hydrolyze the methoxy group portion. Next, after washing the substrate, it is immediately immersed in the aqueous solution of the silane coupling agent, and at room temperature for 1 minute.
Soak for hours. After that, it was washed with running water (ultra pure water), blown with nitrogen gas to be dried, and then heat-fixed in an oven at 120 ° C. for 1 hour.

After cooling, the following formula (II):

[0216]

[Chemical 2]

The substrate was immersed in a 0.3% solution of N- (6-maleimidocaproxy) succinimide (EMCS; compound II) (ethanol: dimethylsulfoxide = 1: 1) at room temperature for 2 hours to obtain EMCS aminosilane. It was reacted with the amino group of the coupling agent. After completion of the reaction, the mixture was washed once with ethanol: dimethyl sulfoxide = 1: 1 and three times with ethanol, and dried by blowing nitrogen gas.

B. Preparation of probe 5'-ATGAACCGGAGGCCCATC-3 '(SEQ ID NO: 1) 3'-TACTTGGCCTCCGGGTAG-5' (SEQ ID NO: 2) having a base sequence which is a base sequence complementary to the above base sequence, and An oligonucleotide (Compound III Bex Co., Ltd.) having a mercapto group (SH group: also known as sulfhydryl group) capable of reacting with a finally purified maleimide group on the surface of the substrate via a linker at the 5 ′ end The probe was used for verification of the examples. Formula (III) below:

[0219]

[Chemical 3]

The mercapto group-introduced oligonucleotide (Compound III) of Example 1 was used as a solvent, that is, 7.5% by weight of glycerin, 7.5% by weight of urea, 7.5% by weight of thiodiglycol, and general formula (IV):

[0221]

[Chemical 4]

In an aqueous solution containing 1 wt% of acetylene alcohol (for example, trade name: acetylenol EH Kawaken Fine Chemicals Co., Ltd.) represented by, the absorbance is 1.0.
To obtain an oligonucleotide solution.

C. Preparation of multiple types of probe 5'-ATGAAC CG GA GG CCCATC-3 '3'-TACTTG GC CT CC GGGTAG-5' is the same as the nucleotide sequence in B above, but in reality, it is a mutation in the nucleotide of the carcinogenesis-related gene p53. It is a region of 18 nucleotides that contains a total of four base portions (underlined) that code for two amino acids with high frequency. The base sequence that is completely complementary to the base sequence of is (that is, the same as the sequence of B above), and the portion corresponding to the base underlined is underlined. 5′-GATGGGN ¹ N ² TCN ³ N ⁴ GTTCAT-3 ′ (SEQ ID NO: 3) has the base at the position underlined as N ¹ ,
It is represented by N ² , N ³ and N ⁴ . These N ¹ , N ² ,
A total of 256 kinds of oligonucleotide probes were synthesized in which N ³ and N ⁴ were substituted with any of the ATGC bases, and a mercapto group was bound to the 5 ′ end in the same manner as in the preparation of the probe of B above.

(Example 1) Production of ZBJ head block for ejecting probe solution By the method described in Japanese Patent Laid-Open No. 06-040037 (silicon substrate thickness is 1 mm), the nozzle having a shape of FIG. 6 and a nozzle of 100 dpi was manufactured. ZBJ heads each formed in a two-dimensional matrix array of 50 vertically and horizontally at a pitch were manufactured. That is, the area where the nozzle is formed is 12.7.
mm × 12.7 mm, and the total number of nozzles is 2500. In this case, the opening diameter of the opening 111 at the tip of the nozzle is 50 μm. This nozzle is designed so that the ejection amount is 24 pl when ejecting normal ink. Further, the original use is the ink flow path to the nozzle. The ink flow paths 113 and 114 are used as probe solution reservoirs in this embodiment. The inner diameter of the solution reservoir 114 is about 200 μm and the depth is about 800 μm. That is, the internal volume of the reservoir 114 is calculated to be about 0.25 μl. The amount of probe ejected from the nozzle at one time is 24 pl.
1 corresponds to the amount of manufacturing an array of about 10,000 sheets.

Example 2 Manufacture of DNA Array Using ZBJ Head The oligonucleotide solution prepared in B of Reference Example was used to prepare the square-shaped 16 × 16 = of the ZBJ head prepared in Example 1.
256 solution reservoirs were dispensed using a microdispenser. Before the final filling with the DNA solution, each solution reservoir and nozzle are washed with a solvent having the above composition in advance, and if necessary, an oligonucleotide for the purpose of making the prepared ZBJ head compatible with the solvent. The solution was washed, and the removal of the liquid by vacuum suction was appropriately repeated.

Then, the oligonucleotide solution was discharged onto the substrate that had been treated to introduce the maleimide group. According to the design specifications of the ZBJ head manufactured in Example 1, the amount of liquid per ejected liquid droplet is 24 pl, and under this ejection condition, the diameter of the dot occupied by one liquid droplet applied on the substrate is used. 7 depending on the viscosity of the solution
It is in the range of 0 to 100 μm. On the other hand, as a matter of course, the ejection pattern, the dot density, etc. are the same as the nozzle arrangement of the manufactured head.

Further, the solvent having the above composition has a high moisturizing property, and is dried and concentrated in the solution reservoir, and the liquid of the oligonucleotide solution applied on the substrate is fixed by the reaction with the substrate surface in the next step. Before, it can be prevented from drying and solidifying.

The substrate coated with the solution of the oligonucleotide (compound III) of the formula (III) in a two-dimensional array was kept at room temperature in a moisturizing chamber having a humidity of 100% for 1 hour, and the mercapto group of the oligonucleotide and the substrate were kept. Reaction with the above maleimide group was carried out. After taking out, the substrate was washed in running water (ultra pure water) for about 30 seconds to remove unreacted oligonucleotide.

Next, with respect to the substrate on which the dots for fixing the above-mentioned oligonucleotides were formed in a two-dimensional array, the surface other than the dots was subjected to a blocking treatment.
M phosphate buffer (pH = 7.0, containing 1M NaCl)
Was immersed in a blocking solution prepared by dissolving BSA (bovine serum albumin Sigma-Aldrich Japan) at a concentration of 2% for 1 hour, washed appropriately with the 50 mM phosphate buffer, and stored in this 50 mM phosphate buffer.

Example 3 Dot Shape Evaluation Model of DNA Array by Hybridization Reaction As a target DNA, a formula (V):

[0231]

[Chemical 5]

The DNA molecule of Example 1, ie, the compound V (purchased from Bex Co., Ltd.) having the sequence shown in B of Reference Example and having tetramethylrhodamine bound to the 5 ′ end as a fluorescent label was used. Hybridization reactions with the probes on the two prepared two-dimensional arrays were performed.

This hybridization reaction was carried out by using the DNA array prepared in Example 2 and a phosphate buffer containing Compound V at a concentration of 5 nM (10 mM phosphate buffer pH =
7.0 ml containing 50 mM NaCl) and 2 ml of
Performed in a Hybripack. The substrate was sealed in a hybrid pack together with the model target DNA solution, and kept at 70 ° C in a constant temperature bath.
To 50 ° C, then cool to 50 ° C, then 1
It was left for 0 hours.

Next, the substrate is taken out from the hybrid pack and washed with a hybridization buffer for the purpose of removing unreacted target DNA. After washing, the substrate was placed on a slide glass in a state of being covered with a buffer solution, covered with a cover glass, and the fluorescence from the fluorescent label was observed. The fluorescence microscope used for this observation was ECLIPSE E80.
0 (Nikon Corporation) with a 20 × objective lens (plan apochromat) and a fluorescent filter (Y-2E / C) set. Images observed by a fluorescence microscope were captured using a CCD camera with an image intensifier (C2400-87 Hamamatsu Photonics KK) and an image processing device (Argus 50 Hamamatsu Photonics KK).

Based on the recorded images, fluorescence was observed for all the dots on which the compound V was fixed, which were formed in a two-dimensional array on the substrate. The average value of the fluorescence intensity was 1750 as the light intensity index of the above measuring device. Further, the average value of the diameters of the dots was calculated from each region emitting the fluorescence, and it was about 100 μm.

(Example 4) 256 prepared in C of Reference Example
A solution of each kind of oligonucleotide probe was prepared in the same manner as in Reference Example A, and was discharged onto a glass substrate in the same manner as in Example 2 and was subjected to reaction bonding to form 256 kinds of DAN probes. A two-dimensionally arrayed DNA probe array was manufactured.

Example 5 Evaluation of Target DNA Selection Characteristics by Hybridization Using the 256 types of DNA probe arrays produced in Example 4, a hybridization reaction was carried out in the same manner as in Example 3 to obtain the objective. Target DN having a base sequence of
The screening characteristics of A were verified.

As model target DNAs, four kinds of DNA having the following nucleotide sequences and having tetramethylrhodamine as a fluorescent label bound to the 5'end were synthesized in the same manner as in Example 3. 5′-ATGAACCGGAGGCCCATC-3 ′ 5′-ATGAAC G GGAGGCCCATC-3 ′ (SEQ ID NO: 4) C → G 5′-ATGAAC GC GAGGCCCATC-3 ′ (SEQ ID NO: 5) C → G, G → C 5′− ATGAAC GC GA A GCCCATC-3 ′ (SEQ ID NO: 6) C → G, G → C, G → A As described above, it is completely complementary to the normal sequence of the p53 gene, and Thus, it is a model of a mutant in which the base sequence is replaced with an underlined base.

Hybridization was carried out for these four kinds of target DNA individually by the same method as in Example 3, and each of the dots of the DNA array produced in Example 4 was examined. Fluorescence was observed only from the dots of the probe having a base sequence completely complementary to the target DNA, and no fluorescence was observed from the other dots. In each of the dots where fluorescence was observed, the observed fluorescence intensity was 1830 for the target DNA, 1270 for the target DNA, 1520 for the target DNA, and 1940 for the target DNA.

Target DNA with some variations
1830 is the average value 1 in Example 3 above.
750 agrees within the statistical error, and the difference in fluorescence intensity of the other three types is in a range that cannot be said to be significant. Therefore, according to the production method of the present invention, a DNA array produced by applying a probe solution in parallel on a substrate using an array-shaped liquid ejecting apparatus can be sufficiently used for quantitative detection of target DNA. It can be seen that the probe amount is fixed with uniformity and reproducibility.

Example 6 An apparatus for manufacturing a probe array having a liquid ejection device having the structure of the embodiment 2 described above was prepared.

Next, the oligonucleotide solution prepared in B of Reference Example was supplied to the liquid reservoir of the liquid ejection device using a microdispenser. Before the final filling with the oligonucleotide solution, each liquid reservoir and nozzle are preliminarily washed with the solvent of the above composition, and if necessary, for the purpose of making the prepared liquid ejection device compatible with the solvent. Washing with the oligonucleotide solution was performed, and the removal of the liquid by vacuum suction was appropriately repeated.

Then, the oligonucleotide solution was discharged onto the substrate prepared by introducing a maleimide group prepared in A of Reference Example. According to the design specifications of the above liquid ejecting apparatus, the amount of liquid per ejected liquid droplet is 24 pl, and under this ejection condition, the diameter of the dot occupied by one liquid droplet applied on the substrate is the viscosity of the solution used. Depending on 70-10
The range is 0 μm.

Further, the solvent having the above composition has a high moisture retention property, and is dried and concentrated in the liquid reservoir, and the liquid of the oligonucleotide solution applied on the substrate is fixed by the reaction with the substrate surface in the next step. Before, it can be prevented from drying and solidifying.

The substrate coated with the oligonucleotide (compound III) of the formula (III) in a two-dimensional array was kept at room temperature for 1 hour in a moisturizing chamber having a humidity of 100%, and the mercapto group of the oligonucleotide and the substrate were kept. Reaction with the above maleimide group was carried out. After taking out, the substrate was washed in running water (ultra pure water) for about 30 seconds to remove unreacted oligonucleotide.

Next, with respect to the substrate on which the dots for fixing the above-mentioned oligonucleotides were formed in a two-dimensional array, the surface other than the dots was subjected to a blocking treatment.
M phosphate buffer (pH = 7.0, containing 1M NaCl)
Was immersed in a blocking solution prepared by dissolving BSA (bovine serum albumin Sigma-Aldrich Japan) at a concentration of 2% for 1 hour, washed appropriately with the 50 mM phosphate buffer, and stored in this 50 mM phosphate buffer.

(DN by hybridization reaction
Evaluation of dot shape of A array) As model target DNA,
The above-mentioned DNA molecule of the formula (V), that is, the compound V (purchased from Bex Co., Ltd.) having the sequence shown in B of Reference Example and having tetramethylrhodamine bound to the 5 ′ as a fluorescent label is used. Then, a hybridization reaction with the probes on the manufactured two-dimensional array was performed.

This hybridization reaction was carried out by using the prepared DNA array and a phosphate buffer containing Compound V at a concentration of 5 nM (10 mM phosphate buffer pH = 7.0, 50).
2 ml (containing mM NaCl). The substrate was sealed in a hybrid pack together with the model target DNA solution and heated to 70 ° C. in a constant temperature bath,
Then, it was cooled to 50 ° C. and left in that state for 10 hours.

Next, the substrate is taken out from the hybrid pack and washed with a hybridization buffer for the purpose of removing unreacted target DNA. After washing, the substrate was placed on a slide glass in a state of being covered with a buffer solution, covered with a cover glass, and the fluorescence from the fluorescent label was observed. The fluorescence microscope used for this observation was ECLIPSE E80.
0 (Nikon Corporation) with a 20 × objective lens (plan apochromat) and a fluorescent filter (Y-2E / C) set. Images observed by a fluorescence microscope were captured using a CCD camera with an image intensifier (C2400-87 Hamamatsu Photonics KK) and an image processing device (Argus 50 Hamamatsu Photonics KK).

Based on the recorded images, fluorescence was observed for all the dots to which the compound V was fixed, which were formed in a two-dimensional array on the substrate. The average value of the fluorescence intensity was 1750 as the light intensity index of the above measuring device. Further, the average value of the diameters of the dots was calculated from each region emitting the fluorescence, and it was about 100 μm.

The oligonucleotide probe synthesized in C of Reference Example was discharged onto a glass substrate using the liquid discharging apparatus having the above-mentioned structure, and was subjected to reaction bonding to give 256
A two-dimensional array of DNA probe arrays consisting of seed DNA probes was manufactured. As described above, the liquid ejecting apparatus used here has 1000 nozzles,
Here, the liquid was ejected using 256 of these nozzles.

(Target DN by hybridization
Evaluation of selection characteristics of A) A hybridization reaction was performed using the 256 types of DNA probe arrays produced, and the selection characteristics of the target DNA having the target nucleotide sequence were verified.

As the model target DNA, four types (having base sequences of, and, respectively) in which tetramethylrhodamine was bound to 5 ′ as the fluorescent label used in Example 5 were used.

[0254] As described above, a model of a mutant which is completely complementary to the normal sequence of the p53 gene, in which replacement with a base underlined in the base sequence occurs. Is.

When the four types of target DNAs of ,,, and were individually hybridized, among the dots of the prepared DNA array, the probe having a base sequence completely complementary to each target DNA was detected. Fluorescence was observed only from the dots and no fluorescence was observed from the other dots. Regarding the fluorescence intensity of the dots where fluorescence was observed, Example 5 was obtained for each target DNA.
Similar results were obtained.

In other words, although there are some variations,
The difference in the fluorescence intensity was in a range that was not significant. Therefore, according to the production method of the present invention, a DNA array produced by applying a probe solution in parallel on a substrate using an array-shaped liquid ejecting apparatus can be sufficiently used for quantitative detection of target DNA. It can be seen that the probe amount is fixed with uniformity and reproducibility.

[0257]

EFFECT OF THE INVENTION In the method for producing a probe array of the present invention, a plurality of types of probes that have been separately synthesized, purified and confirmed are
Using a liquid ejecting apparatus or a liquid ejecting apparatus having a solution reservoir for accommodating each probe solution and a number of solution ejecting nozzles connected to the solution reservoir corresponding to the plurality of kinds of probes, each probe solution has a constant liquid amount. Since it is manufactured into a probe array by discharging and applying it in an array on the substrate with, the uniformity of the probe amount contained in the dots of each probe is high, and when reproducing multiple plates, reproduction It also has the advantage that it can be kept sufficiently high.

In particular, when an array-shaped liquid ejecting apparatus is used, the ink jet head itself can be formed into a shape having a solution reservoir and nozzles that are highly integrated,
Further, by miniaturizing the nozzle size and reducing the amount of the ejected liquid, it is possible to efficiently apply the dots to the substrate at a high density by parallel ejection.

According to the present invention, the liquid ejecting apparatus or the unit chips of the liquid ejecting apparatus are formed into a high-density array, so that a larger number of probe species constituting a two-dimensional array on the solid-phase substrate can be obtained. It is possible to easily achieve high density, high density of dots associated therewith, and reduction of occupied area of individual dots.

[0260]

[Sequence list] SEQUENCE LISTING <110> Canon INC. <120> A method of preparing a probe array and a device used therefor <130> 4396102 <150> JP 2000-284046 <151> 2000-09-19 <160> 6 <210> 1 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Base Oligonucleotide for preparation of a probe <400> 1 atgaaccgga ggcccatc 18 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Oligonucleotide probe for hybridization assay <400> 2 tacttggcct ccgggtag <210> 3 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Oligonucleotide probe for hybridization assay.n = a, t, g or c. <400> 3 gatgggnntc nngttcat 18 <210> 4 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Oligonucleotide probe for hybridization assay. <400> 4 atgaacggga ggcccatc 18 <210> 5 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Oligonucleotide probe for hybridization assay. <400> 5 atgaacgcga ggcccatc <210> 6 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Oligonucleotide probe for hybridization assay. <400> 6 atgaacgcga agcccatc

[Brief description of drawings]

FIG. 1 is a perspective view showing an example of a separated two-part bubble jet print head.

FIG. 2 is a perspective view showing an example of a separated two-part bubble jet print head.

FIG. 3 is a perspective view showing an integrally formed ZBJ chip.

FIG. 4 is a cutaway perspective view showing a first example of a ZBJ head.

FIG. 5 is a cutaway perspective view showing a second example of the ZBJ head.

6 (A), (B), (C) and (D) are Z
It is explanatory drawing which shows an example of the etching process used when forming a BJ nozzle.

FIG. 7 is a schematic diagram showing an example of possible arrangement of heater elements in a ZBJ substrate.

FIG. 8 is a schematic diagram showing an example of possible arrangement of heater elements in a ZBJ substrate.

FIG. 9 is a schematic diagram showing an example of possible arrangement of heater elements in a ZBJ substrate.

FIG. 10 is a schematic view showing another example of the heater structure of the heater element in the ZBJ substrate.

FIG. 11 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 12 is a cross-sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 13 is a cross-sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 14 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 15 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 16 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 17 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 18 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 19 is a sectional view showing an example of a nozzle shape of a ZBJ head.

FIG. 20 is a cross-sectional view showing an example of a nozzle shape of a ZBJ head.

21A to 21D are cross-sectional views showing how ink is ejected from nozzles of a ZBJ chip.

22A to 22D are plan views showing how ink is ejected from the nozzles of the ZBJ chip.

FIG. 23 is an explanatory diagram showing transfer of heat generated in a heater element in a ZBJ chip.

FIG. 24 is an explanatory diagram showing the transfer of heat generated in the heater element in the ZBJ chip.

FIG. 25 is an explanatory diagram showing transfer of heat generated in a heater element in a ZBJ chip.

FIG. 26 is an explanatory diagram showing transfer of heat generated in a heater element in a ZBJ chip.

FIG. 27 is an explanatory diagram showing transfer of heat generated in a heater element in a ZBJ chip.

FIG. 28 is an exploded perspective view showing the arrangement of a ZBJ printhead including tips, membrane filters and ink channel extrusions.

FIG. 29 is a cross-sectional view showing another structure of the ink channel extruded body.

FIG. 30 is a schematic view of a liquid ejection apparatus having a two-dimensional array nozzle arrangement used in the present invention.

FIG. 31 is a schematic view of a semiconductor chip constituting a liquid jet apparatus of the thermal jet method according to the present invention.

32 is an enlarged view of the vicinity of the nozzle in FIG. 31. FIG.

33 is a cross-sectional view taken along the line AA ′ of FIG.

34 is a cross-sectional view taken along the line BB ′ of FIG.

FIG. 35 is a schematic view of a back surface of a semiconductor chip (corresponding to the back surface of the semiconductor chip shown in FIG. 1) that constitutes a liquid ejection apparatus of the thermal jet system.

FIG. 36 is a schematic diagram for explaining the shape of the liquid supply port (reservoir) of the semiconductor chip that constitutes the liquid jet apparatus of the thermal jet system.

FIG. 37 is a schematic diagram for explaining ejection of a liquid by a liquid ejection device using a thermal jet method.

FIG. 38 is a schematic view of a liquid ejection device.

FIG. 39 is a schematic diagram of a liquid ejection device.

FIG. 40 is a schematic view of another embodiment of the liquid ejection device.

FIG. 41 is a schematic view of the structure of a probe array manufacturing apparatus.

FIG. 42 is a schematic diagram for explaining an example of a method for applying a probe solution using a liquid ejecting apparatus by a thermal jet method.

FIG. 43 is a diagram showing an example of a nozzle formation surface of a liquid ejection device.

FIG. 44 is a schematic diagram for explaining another example of a method for applying a probe solution using a liquid ejecting apparatus using a thermal jet method.

[Explanation of symbols]

1 Substrate 2 Heater Element 3 First Wiring 4 Second Wiring 5 Pad 6 Nozzle 7 Channel 8 Supply Port (Liquid Reservoir) 9 Contour of Maximum Outer Edge of Supply Port 10 Insulating Film 11 Protective Film 12 Nozzle Material 13 Cavitation Resistant Film 100 ZBJ chip (bubble jet printing device) 106 Ink 108 Ink drop 110 Nozzle path (nozzle) 113,487 Barrel 114,489 Nozzle channel (passage) 115,488 Heat chamber (heat chamber) 120,440 Heater (heater) Means) 121,441 Main heater 122,443 Redundant heater 130 Substrate 132 Thermal insulation layer (high operating temperature part) 140 Heat shunt (heat conductor) 160 Heater driver 201,202 Power busbar 205 Filter (membrane) 210 Channel extruded body ( Ink supply means 220 Application target (applied Cement medium)

─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Tomohiro Suzuki 3-30-2 Shimomaruko, Ota-ku, Tokyo Canon Inc. (56) Reference JP-A-6-40037 (JP, A) JP-A-11 -187900 (JP, A) International Publication 98/036833 (WO, A1) (58) Fields investigated (Int.Cl. ⁷ , DB name) G01N 33/53 G01N 37/00 C12M 1/00 C12N 15/09

Claims (36)

(57) [Claims] 1. A method of manufacturing a probe carrier having a plurality of types of probes at different positions on a carrier, comprising at least a liquid reservoir for storing a liquid containing the probe and a liquid ejection nozzle communicating with the liquid reservoir. A liquid having a number corresponding to a plurality of types of probes, the reservoir and the liquid ejection nozzle are formed on the same substrate, and the reservoir has a supply port for supplying different liquids, respectively. A step of preparing an ejection device; a step of relatively aligning the plurality of liquid ejection nozzles with the carrier; and a step of preparing a liquid containing the probes from different positions on the carrier from the liquid ejection nozzle. A step of ejecting the probe carrier. 2. The method for producing a probe carrier according to claim 1, wherein the probe is at least a part of a nucleic acid. 3. The method of manufacturing a probe carrier according to claim 1, wherein in the liquid ejection device, the plurality of liquid ejection nozzles are linear or are opened in a vertical direction and a horizontal direction. 4. The liquid reservoir is located on the opposite side of the liquid discharge nozzle from the liquid discharge direction.
4. The method for producing a probe carrier according to any one of 3 to 3. 5. The method for manufacturing a probe carrier according to claim 1, wherein a separate liquid storage portion is further connected to and connected to the liquid reservoir. 6. The probe carrier according to claim 1, wherein the liquid ejection device includes a thermal energy generator for applying thermal energy to the liquid in the nozzle to eject the liquid. Manufacturing method. 7. The liquid ejecting apparatus according to claim 6, wherein the liquid is ejected by the bubbles generated by applying thermal energy to the liquid communicating with the outside air. 8. The method of manufacturing a probe carrier according to claim 1, wherein the liquid is applied onto the carrier by using the liquid ejecting device at the same density as the nozzle density of the liquid ejecting device. 9. The liquid containing the probe is made to have a higher density than the nozzle density of the liquid ejecting apparatus by performing a relative movement of the liquid ejecting apparatus and the carrier a plurality of times with respect to one carrier. The method for producing a probe carrier according to claim 1, wherein the probe carrier is provided on the carrier. 10. A liquid ejecting apparatus for producing a probe carrier having a plurality of types of probes at different positions on a carrier, the liquid ejecting device communicating with the liquid reservoir for accommodating the liquid containing the probe. A plurality of nozzles for at least the plurality of types of probes are provided, the liquid reservoir and the liquid ejection nozzle are formed on the same substrate, and supply ports for supplying different liquids to the reservoirs, respectively. A liquid ejecting apparatus comprising: 11. The liquid ejection apparatus according to claim 10, wherein the probe is at least a part of nucleic acid. 12. The liquid ejecting apparatus according to claim 10, wherein the plurality of liquid ejecting nozzles are opened linearly or in a vertical direction and a horizontal direction. 13. The liquid ejecting apparatus according to claim 10, wherein the liquid reservoir is located on the opposite side of the liquid ejecting nozzle from the liquid ejecting direction. 14. The liquid reservoir is further connected to and connected to the liquid reservoir, which is a separate body.
4. The liquid ejection device according to any one of 3 above. 15. The liquid ejection apparatus according to claim 10, further comprising a thermal energy generator for applying thermal energy to the liquid in the nozzle to eject the liquid. 16. The liquid ejecting apparatus according to claim 15, wherein the liquid is ejected when bubbles generated by applying thermal energy to the liquid communicate with the outside air. 17. The liquid ejection device according to claim 10, wherein the liquid ejection device is a device for applying the liquid onto the carrier at the same density as the nozzle density of the liquid ejection device. apparatus. 18. The liquid ejecting apparatus makes the liquid containing the probe have a density higher than the nozzle density of the liquid ejecting apparatus by performing relative movement of the carrier a plurality of times with respect to one of the carriers. 17. The liquid ejection device according to claim 10, wherein the liquid ejection device is a device for applying on the carrier. 19. A probe carrier manufacturing apparatus comprising the liquid ejection device according to claim 10. 20. An apparatus for producing a probe carrier having a plurality of types of probes on different positions on a carrier, wherein at least a liquid reservoir for storing a liquid containing the probe and a liquid ejection nozzle communicating with the liquid reservoir are provided. A liquid ejector having a number corresponding to a plurality of types of probes, the liquid reservoir and the liquid ejection nozzle are formed on the same substrate, and each of the reservoirs has a supply port for supplying a different liquid. Device,
A probe carrier manufacturing apparatus having a positioning means for relatively positioning the plurality of liquid discharge nozzles and the carrier. 21. An integrated flat plate-shaped substrate having a laminated layer structure, a plurality of liquid ejection nozzles formed and arranged in a two-dimensional array on one surface of the substrate, and the other surface of the substrate. A liquid reservoir formed in a two-dimensional array in communication with the plurality of liquid ejection nozzles, and a liquid formed in a two-dimensional array corresponding to each of the liquid ejection nozzles. An energy generating element for discharge,
And a supply port for supplying a different liquid to each of the reservoirs provided by opening on the surface on which the liquid reservoir is arranged. 22. The liquid ejection apparatus according to claim 21, wherein the nozzles, the reservoirs, and the energy generating elements are formed and arranged in at least 5 rows and 8 columns. 23. The liquid ejecting apparatus according to claim 21, wherein each of the liquid reservoirs individually includes a volume increasing reservoir connected thereto. 24. Each of the volume increasing reservoirs is formed as an integral plate in an arrangement corresponding to a two-dimensional array of the liquid reservoirs, and the plate is a surface of the substrate on which the liquid reservoirs are formed. 24. The liquid ejecting apparatus according to claim 23, wherein the liquid reservoirs and the volume increasing reservoirs corresponding to the reservoir groups are stacked and fixed in a positional relationship in which the liquid reservoirs are connected to each other. 25. The liquid ejecting apparatus according to claim 23, wherein an interval between adjacent reservoir centers of the liquid reservoir and an interval between adjacent reservoir centers of the volume increasing reservoir are the same. 26. The monolithic flat substrate is provided on a surface of a silicon substrate, an insulating film such as a silicon oxide film or a silicon nitride film, a metal film such as AL or Ta, an energy generating element material,
The liquid ejection device according to any one of claims 21 to 25, which is a substrate in which nozzle materials are laminated. 27. The shape of the liquid reservoir is gradually narrowed from the opening side toward the nozzle side.
27. The liquid ejection device according to any one of items 26 to 26. 28. The liquid ejecting apparatus according to claim 27, wherein the liquid reservoir has a truncated pyramid shape. 29. The liquid ejection device according to claim 23, wherein the volume increasing liquid reservoir has a cylindrical shape. 30. The liquid ejecting apparatus according to claim 21, wherein an interval between adjacent nozzle centers of the liquid nozzle and an interval between adjacent reservoir centers of the reservoir are the same. 31. The liquid ejecting apparatus according to claim 22, wherein the nozzles arranged in the two-dimensional array have uniform row intervals and column intervals in the substrate. 32. The liquid ejecting apparatus according to claim 22, wherein the row interval and the column interval are the same in the substrate. 33. The liquid ejecting apparatus according to claim 22, wherein the energy generating element is a piezo element. 34. The liquid ejecting apparatus according to claim 22, wherein the energy generating element is a heat generating element. 35. A liquid discharge part of the liquid discharge device is formed on the substrate, an insulating region provided at a predetermined position on the substrate, a heat generating element formed on the insulating region, and a heat generating element on the heat generating element. Protective film, anti-cavitation film formed on the protective film, flow path for holding liquid above the anti-cavitation film, nozzle for discharging liquid in communication with the liquid flow path, nozzle of the substrate The heat generating element element is composed of a supply port functioning as a liquid reservoir communicating with the flow path provided on the back side with respect to the side where the heat generation element is formed. Both ends thereof are connected to each other, the first and second wirings are respectively connected to pads for inputting an electric signal from the outside of the substrate, and the heat generating element is connected to an electric signal applied to each pad. 35. Driven according to claim 34. Liquid ejector. 36. The liquid ejection apparatus according to claim 21, wherein the nozzle is formed by etching the laminated layer structure.