JP2002116205A - Liquid discharge device as well as method and apparatus for manufacture of microarray using it - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a liquid discharge device which can discharge a very small liquid and whose nozzle is hard to clog. SOLUTION: The liquid discharge device is provided with a liquid holding member 34 comprising a discharge port and a drive means 32 to generate a discharge pressure which discharges a very small amount of the liquid held in the member 34 from the discharge port. The discharge port is constituted of an opening 39 which faces the outside air and a very small hole 41 whose cross-sectional area is smaller than that of the opening 39.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention includes a liquid ejecting apparatus for ejecting a small amount of liquid such as a reagent and a chemical solution, and a probe capable of specifically binding to a target substance using the above liquid ejecting apparatus. The present invention relates to a microarray manufacturing method for manufacturing a microarray formed by discharging a very small amount of liquid onto a substrate, and a microarray manufacturing apparatus for performing the microarray manufacturing method.

[0002]

2. Description of the Related Art In a blood analyzer, a testing apparatus for a genetic test, a drug testing test, and the like, it is desired to reduce the minimum discharge amount of a liquid discharge system in order to reduce running costs and improve throughput.

Conventionally, a syringe piston pump has been used as a liquid ejection technique for such an inspection apparatus. However, the minimum discharge amount of the syringe piston pump is about 2 μL, and there has been a problem that the discharge accuracy is extremely deteriorated if a small amount of liquid is to be discharged.

As a solution to this problem, for example, Japanese Patent Application Laid-Open No. Hei 10-114394 discloses a microfluidic processing apparatus capable of forming discontinuous droplets of less than 1 nanoliter and having a substantially reproducible size. Is disclosed.

FIG. 13 shows the configuration of such a microfluidic processing apparatus. The microfluidic processing apparatus 10 includes a microdispenser 16 using a piezoelectric transducer attached to a glass capillary, filling the microdispenser 16 with a transfer fluid, and aspirating the transfer fluid from the microdispenser 16 to remove a system fluid. It includes a positive displacement pump 12 that controls the pressure and flushes the microdispenser 16 between transfers, and a pressure sensor 14 that measures the pressure of the system fluid and emits a corresponding electrical signal.

[0006] The microdispenser 16 has a piezoelectric ceramic tube 60 connected to a glass capillary tube 62 as shown in FIG. 14. The piezoelectric ceramic tube 60 has an inner electrode 66 and an outer electrode 68 for applying an analog voltage pulse for contracting the tube.

When discharging the transfer fluid from the microdispenser 16, an analog voltage pulse is applied to the piezoelectric ceramic tube 60 to contract the piezoelectric ceramic tube 60,
This deforms the glass capillary 62 to form a pressure wave that travels through the transfer fluid and reaches the nozzle 63 of the microdispenser 16, and this pressure wave causes one droplet of the transfer fluid from the nozzle 63 to have an extremely high acceleration. It is made to inject at. In this prior art, for example, 5 picoliter droplets have been demonstrated.

[0008]

SUMMARY OF THE INVENTION The above-mentioned Japanese Patent Application Laid-open No. Hei 10-1
The microfluidic processing apparatus disclosed in Japanese Patent No. 14394 is an excellent technique capable of discharging a very small amount of liquid with high accuracy, but has a problem that nozzle clogging is likely to occur on the other hand.

That is, in a liquid ejection apparatus utilizing an ink jet system, it is necessary to reduce the nozzle diameter in order to generate minute droplets.
In order to generate droplets of 1 nanoliter or less, the diameter of the nozzle needs to be 0.1 mm or less at most. However, as the nozzle diameter is reduced, nozzle clogging is more likely to occur.

On the other hand, liquid samples handled by a genetic tester or a biochemical analyzer often contain cells and proteins. According to the experience of the present inventors, these liquid samples have a nozzle diameter of 0. It was found that when the thickness was 0.1 mm or less, nozzle clogging was likely to occur. Particularly, in a genetic tester used for research purposes, since the setup of the reaction container and the suction sample container is frequently changed, the sample in the nozzle may be dried and clogged during this time.

When nozzle clogging occurs as described above,
It is necessary to remove the clogged substance by immersing the nozzle in washing water or the like, or by supplying a liquid into the chamber to increase the pressure. For this reason, if nozzle clogging frequently occurs, the throughput will be reduced accordingly.

Accordingly, a first object of the present invention, which has been made in view of the above points, is to provide a liquid discharge apparatus which can discharge a minute fluid and is less likely to cause nozzle clogging.

Further, a second object of the present invention is to provide a microarray in which a very small amount of liquid containing a probe capable of specifically binding to a target substance is discharged onto a substrate using the above-described liquid discharging apparatus. An object of the present invention is to provide a microarray manufacturing method that can be manufactured easily and efficiently.

Further, a third object of the present invention is to provide a microarray manufacturing apparatus which can execute the above microarray manufacturing method with a simple configuration.

[0015]

According to a first aspect of the present invention, there is provided a liquid ejecting apparatus, comprising: a liquid holding member having a discharge port; and a liquid holding member for discharging the liquid held by the liquid holding member. A drive means for generating a discharge pressure for discharging a very small amount from an outlet, wherein the discharge port has an opening facing the outside atmosphere, and a fine hole having a smaller cross-sectional area than the opening. It is characterized by comprising.

According to the first aspect of the present invention, when the ejection pressure is applied to the liquid held by the liquid holding member by the driving means at the time of ejecting the liquid, the pressure of the liquid in the minute holes increases instantaneously, and A very small amount of liquid is discharged from the discharge port against the surface tension at the time, and the discharge amount at that time is determined not by the diameter of the opening but by the diameter of the minute hole. Further, since the liquid faces the atmosphere at the opening having a large cross-sectional area, clogging hardly occurs. Therefore, clogging at the discharge port can be effectively prevented, and a very small amount of liquid can be reliably discharged.

According to a second aspect of the present invention, in the liquid ejecting apparatus according to the first aspect, the driving means is configured to move the liquid holding member forward and backward along the ejection direction. It is.

According to the second aspect of the invention, by moving the liquid holding member in the direction opposite to the discharge direction, the pressure of the liquid in the micropores can be instantaneously increased by the inertial force acting on the liquid, The discharge pressure makes it possible to reliably discharge a very small amount of liquid from the discharge port against the surface tension at the opening.

According to a third aspect of the present invention, in the liquid ejecting apparatus according to the second aspect, the space between the opening of the ejection port and the minute hole is formed in a tapered shape, and the liquid holding device is provided by the driving means. The discharge pressure is generated by moving the member in the discharge direction and then in the opposite direction.

According to the third aspect of the present invention, the liquid held in the opening moves toward the inside of the liquid holding member from the minute holes along the tapered portion by the movement of the liquid holding member in the ejection direction before the ejection. Since the liquid is pushed back, the liquid can be discharged without attenuating the discharge flow velocity as compared with the case where the liquid is held in the opening at the time of discharge, and the discharge efficiency can be improved.

According to a fourth aspect of the present invention, in the liquid discharging apparatus according to the first aspect, the driving means is configured to change a volume of the liquid holding member.

According to the fourth aspect of the present invention, since the volume of the liquid holding member is changed, it is possible to easily generate a desired discharge pressure.

According to a fifth aspect of the present invention, in the liquid ejecting apparatus according to any one of the first to fourth aspects, the driving means includes a piezoelectric element.

According to the fifth aspect of the present invention, since the driving means is constituted by using the piezoelectric element, the driving means can be constituted simply and inexpensively.

According to a sixth aspect of the present invention, there is provided a microarray manufacturing method which achieves the second object, using the liquid ejecting apparatus according to any one of the first to fifth aspects, wherein the liquid ejecting apparatus applies a target substance to the liquid. A microarray is manufactured by discharging a small amount of liquid containing a probe capable of specifically binding to the substrate.

According to the invention of claim 6, claims 1 to
Since the microarray is manufactured using the liquid ejection device according to any one of the above items 5, it is possible to easily and efficiently manufacture the microarray.

According to a seventh aspect of the present invention, which achieves the third object, a microarray is manufactured by discharging a small amount of a liquid containing a probe capable of specifically binding to a target substance onto a substrate. A device,
A liquid ejection device according to any one of claims 1 to 5,
Relative moving means for relatively moving the substrate and the liquid holding member in a plane perpendicular to the direction in which the liquid is ejected from the liquid holding member provided in the liquid ejection device. Things.

According to the invention according to claim 7, claims 1 to 1 are provided.
The liquid holding member of the liquid ejecting apparatus of No. 5 is used, and the liquid holding member and the substrate are relatively moved by a relative moving means in a plane orthogonal to the direction in which the liquid is ejected from the liquid holding member. Thus, it is possible to easily and efficiently manufacture a microarray.

[0029]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the present invention will be described below with reference to FIGS.

1 to 4 show a first embodiment of the present invention. FIG. 1 is a schematic structural view of a liquid discharge device, FIG. 2 is a sectional view of the liquid discharge head shown in FIG. 1, and FIG. FIG. 4 is a diagram showing a drive voltage waveform applied to the laminated piezoelectric element shown in FIG. 2;
(A)-(e) is a figure for demonstrating the discharge operation of a test sample.

In FIG. 1, the liquid discharge head 20 is supported by a movable transfer member (not shown), and is movably disposed above the test sample container 21, the washing tank 22, and the reaction container 23. One end of the liquid ejection head 20 is connected to a syringe piston pump 27 via a pipe 25 made of Teflon (trade name) or the like. The syringe piston pump 27 is connected to another pipe 25 leading to a liquid supply tank 28 in addition to the above-described pipe 25, and is sequentially connected to the liquid supply tank 28 via a solenoid valve 29 and a water supply pump 30.

The piston of the syringe piston pump 27 reciprocates in the direction of the arrow by a linear reciprocating actuator such as a stepping motor and a gear rack pinion (not shown). The liquid discharge head 20, the syringe piston pump 27, and the liquid supply tank 28
They are installed at approximately the same height.

As shown in detail in FIG. 2, the liquid discharge head 20 includes a flow path member (liquid holding member) 34 and a laminated piezoelectric element (drive means) 32 for moving the flow path member 34 forward and backward. Be composed. One end of the multilayer piezoelectric element 32 in the displacement direction is fixed to the gantry 33, and the other end is fixed to the flow path member 34.

The flow path member 34 has a flow path 35 for holding a liquid therein, and an inlet 36 for introducing the liquid into the flow path 35. The flow path 35 includes a straight portion 37, a tapered portion 38, and an opening 39. Opening 39
Has an opening 40 having a depth of 0.5 mm to 1 mm, a diameter of 0.2 mm to 1 mm communicating with the outside atmosphere at one end, and a diameter of 0.03 mm to 0.1 m at the center at the other end.
m small holes 41 are provided.

The tapered portion 38 is formed so that the cross-sectional area of the flow path increases from the minute hole 41 toward the straight portion 37.
It is provided with a taper angle of degrees to 45 degrees. The straight portion 37 has a diameter of, for example, 2 mm to 10 mm and a length of 5 mm.
It is formed with a size of １５15 mm.

The discharge direction of the flow path member 34 is parallel to the forward and backward movement directions of the laminated piezoelectric element 32, and a water-repellent layer made of a fluorine-based material, which is a low surface energy substance, is provided on the lower end surface and the outer peripheral surface. Have been.

The space between the laminated piezoelectric element 32 and the flow path 35 is formed as a rigid body. In addition, since one end of the multilayer piezoelectric element 32 is fixed to the gantry 33, the entire flow path member 34 moves up and down with the displacement of the multilayer piezoelectric element 32. A voltage of a desired waveform is applied to the laminated piezoelectric element 32 from a drive circuit (not shown) via a lead wire or a flexible substrate.

Next, the operation of this embodiment will be described. First, the liquid discharge head 20 is moved above the cleaning tank 22, and the tip of the liquid discharge head 20 is immersed in the cleaning tank 22.
At 0, the cleaning water in the liquid supply tank 28 is sent to the liquid ejection head 20, and the inner peripheral surface of the flow channel 35, the outer peripheral surface of the tip of the flow channel member 34, and the lower end surface are cleaned with the cleaning water.

While the washing water is flowing, the syringe piston pump 27 moves to the middle point, after which the solenoid valve 29 is closed and the flow path 35 is filled with the washing water. Next, the liquid discharge head 20 moves above the cleaning tank 22, the syringe piston pump 27 moves downward by a predetermined amount, and sucks a predetermined amount of air into the flow path 35 through the opening 40.

Thereafter, the liquid discharge head 20 moves to the test sample container 21 and immerses the tip of the liquid discharge head 20 in the test sample by 1 mm to 2 mm, and the piston of the syringe piston pump 27 moves a predetermined amount downward. Then, the test sample is sucked into the channel 35. The test sample sucked into the flow path 35 is separated from the washing water in the air layer.

When the suction of a predetermined amount of the test sample is completed, the liquid discharge head 20 moves up above the test sample container 21. At this time, since a water-repellent layer made of a fluorine-based material is formed on the end surface and the outer peripheral surface of the leading end of the liquid ejection head 20, the test sample does not adhere to these portions.
Thereafter, the liquid discharge head 20 moves above the reaction container 23 and discharges the test sample into the reaction container 23.

The discharge operation of the test sample will be described in more detail with reference to FIGS. In the present embodiment, the drive voltage E applied to the laminated piezoelectric element 32 is
As shown in (2), the value is continuously increased from E0 to + E1 during the period from t1 to t2, and is immediately decreased to E0 immediately after t2.

Here, as shown in FIG. 4A, t <t
Assuming that the position of the end of the flow path member 34 when E = E0 is A, the flow path member 34 slowly increases as the voltage E increases at t1 <t <t2, as shown in FIG. Descend. At t = t2, the voltage +
A displacement corresponding to E1 occurs, and the front end of the flow path member 34 is
It reaches the position B as shown in FIG.

Thereafter, immediately after t = t2, the voltage E instantaneously changes to E.
0, the flow path member 3 as shown in FIG.
4 also sharply rises with a decrease in the voltage, and when t> t2, the tip position returns to the position A as shown in FIG.

At this time, that is, when the flow path member 34 is
When returning from the state shown in FIG. 4C to the state shown in FIG. 4E, a downward inertial force acts on the test sample in the flow path 35, and the test samples in the flow path 35 are simultaneously and instantaneously. Then, the pressure at the tip of the tapered portion 38 rises instantaneously due to this flow. On the other hand, in the opening 39, since all the end faces in the moving direction of the fluid are open to the outside atmosphere, the pressure of the test sample held here does not increase.

Accordingly, a pressure difference is generated between the test sample in the tapered portion 38 and the test sample in the opening 39, and the pressure difference causes the small hole 41 at the tip of the tapered portion 38 to open the opening 3.
A flow toward 9 occurs. The flow generated here goes straight through the sample while reducing the flow velocity due to the viscous resistance of the test sample in the opening 39, whereby the diameter of the minute hole 41 in the test sample held in the opening 39 is reduced. A corresponding portion of the test sample is extruded and discharged from the opening 40 to the outside.

Here, the amount of the test liquid to be discharged depends on the taper angle of the taper portion 38, the diameter of the minute hole 41, the depth of the opening 39, the driving voltage waveform applied to the laminated piezoelectric element 32, and the physical properties of the test sample. It depends on the value and so on.
It is between 01 nanoliter and 0.3 microliter.

After the desired test sample is discharged, the liquid discharge head 20 is moved to the cleaning tank 22 again, the cleaning water is supplied to the flow path 35, and the test sample remaining in the flow path 35 is discharged. The inner surface of the channel 35 and the outer peripheral surface of the tip of the channel member 34 are cleaned. Thereafter, a desired amount of the test sample is sequentially discharged to the reaction container 23 by repeating the above-described cleaning operation and the suction / discharge operation of the test sample.

According to the present embodiment, the discharge amount depends on the taper angle of the tapered portion 38, the diameter of the minute hole 41, or the opening 39.
And the drive voltage waveform, the physical properties of the test sample, and the like, and the diameter of the opening 40 does not influence. Therefore, the diameter of the opening 40, which dries in contact with the outside air and easily causes clogging of the nozzle, can be increased without lowering the discharge accuracy. Can be reliably discharged.

In this embodiment, the flow path 35 is provided with the tapered portion 38. However, the shape of the flow path 35 is not limited to this. For example, the tapered portion as shown in the sectional view of FIG. The same effect can be obtained even in the case of no. Further, in the above-described embodiment, the test sample is discharged by lowering the flow path member 34 slowly and then steeply raising the flow path member 34 by the laminated piezoelectric element 32. For example, in FIG. Is rapidly lowered from the state at the position A to the position B and stopped, thereby discharging a very small amount of the test sample, and then slowly returning to the position A. Alternatively, in FIG. By setting the state at the position B as the home position and rapidly raising the position from the position B to the position A, a very small amount of the test sample can be discharged, and thereafter, the position can be slowly returned to the position B.

Further, in the above-described embodiment, the flow path member 34 is reciprocated in the discharge direction by the laminated piezoelectric element 32. For example, as shown in a sectional view of FIG. A piezoelectric element (driving means) 80 is provided via 81, and by driving the piezoelectric element 80 to reduce the volume of the flow path 35, pressure is generated in the test sample to discharge a small amount of the test sample. Can also be configured. In this case, the same effect can be obtained.

7 to 9 show a second embodiment of the present invention. FIG. 7 is a sectional view of a liquid discharge head, and FIG.
Diagram showing a drive voltage waveform applied to the laminated piezoelectric element shown in FIG.
FIGS. 9A to 9E are diagrams for explaining the discharge operation of the test sample.

This embodiment is the same as the first embodiment except for the configuration and the ejection operation of the liquid ejection head.
The same components are denoted by the same reference numerals, and description thereof is omitted. FIG.
As shown in FIG.
2 is a flow path member (liquid holding member) 43 and a laminated piezoelectric element (driving means) for moving the flow path member 43 forward and backward.
32. One end of the laminated piezoelectric element 32 in the displacement direction is fixed to the gantry 33, and the other end is
It is fixed to 3.

The flow path member 43 includes a flow path 44 for holding a liquid therein, and an inlet 4 for introducing a liquid from the outside to the flow path 44.
5, and a discharge opening 46 for discharging the liquid from the flow path 44 to the outside. The diameter of the discharge opening 46 facing the external atmosphere is, for example, 0.2 mm to 1 mm.

The flow path 44 includes a straight portion 47, a first tapered portion 48, and a second tapered portion 49. One end of the second tapered portion 49 communicates with the discharge opening 46, and the other end communicates with one end of the first tapered portion 48.
The second taper portion 49 is provided so that the cross-sectional area of the flow path decreases from the discharge opening portion 46 to the first taper portion 48, and the taper angle is 20 degrees to 60 degrees. Also, the first
Is provided so that the cross-sectional area of the flow path increases from one end of the second tapered portion 49 toward the straight portion 47, and the taper angle is 20 degrees to 60 degrees.

The first tapered portion 48 and the second tapered portion 49
The flow path diameter at the boundary portion (micropores) 50, i.e., at the position where the flow path cross-sectional area is the narrowest, is, for example, 0.03 mm to 0.
1 mm. The straight portion 47 has, for example, a width of 2 mm or more.
10 mm, length 5 mm to 15 mm.

The discharge direction of the flow path member 43 is parallel to the moving direction of the multilayer piezoelectric element 32, and a water-repellent layer made of a fluorine-based material which is a low surface energy substance is provided on the lower end surface and the outer peripheral surface. Have been.

The space between the laminated piezoelectric element 32 and the flow path 44 is formed as a rigid body. Further, since one end of the laminated piezoelectric element 32 is fixed to the gantry 33, the entire flow path member 34 moves forward and backward with the displacement of the laminated piezoelectric element 32 without changing the volume. ing. A voltage of a desired waveform is applied to the laminated piezoelectric element 32 from a drive circuit (not shown) via a lead wire or a flexible substrate. Further, the liquid ejection head 42 is provided with cleaning water supply means having a pipe 25, a syringe piston pump 27, a liquid supply tank 28, a solenoid valve 29, and a water supply pump 30, as in the first embodiment.

Next, the operation of this embodiment will be described. Also in the present embodiment, similarly to the first embodiment, after cleaning the flow path 44, air and the test sample are sequentially sucked into the flow path 44, and then the liquid discharge head 42 is connected to the reaction vessel 23. It moves upward to perform a discharge operation of the test sample.

The operation of discharging the test sample will be described in more detail with reference to FIGS. In the present embodiment, the drive voltage E applied to the laminated piezoelectric element 32 is
As shown in the figure, the voltage is rapidly increased from E0 to + E1 from t1 to t2, and the voltage + E1 is maintained for a short time from t2 to t3, and then rapidly changed from + E1 to -E1 from t3 to t4. After that, between t4 and t5-
Gradually decrease from E1 to E0.

Here, t <t1 and E =
From the initial state of E0, the laminated piezoelectric element 32 is moved from t1 to t.
When a voltage that sharply increases from E0 to + E1 is applied in the polarization direction during the period 2, the laminated piezoelectric element 32 rapidly expands and lowers the flow path member 43. When the flow path member 43 suddenly descends in this manner, the test sample in the flow path 44 has
(B), an upward inertial force acts so as to indicate a state of t1 <t <t2, whereby the test sample inside the second taper portion 49 moves upward and moves inside the first taper portion 48. At t = t3, as shown in FIG. 9C, all the test samples in the second tapered portion 49 are removed from the first tapered portion 48.
Flows into

Thereafter, when the voltage applied to the laminated piezoelectric element 32 is rapidly changed from + E1 to -E1 between t3 and t4, the laminated piezoelectric element 32 is rapidly reduced and
3 is raised. When the flow path member 43 rises sharply in this way, the test sample in the flow path 44 is accompanied by FIG.
(D), as shown in the state of t3 <t <t4, a flow in the same direction occurs due to an inertial force acting in the downward direction, whereby a very small amount of the test sample from the tip of the first taper portion 48 is generated. Is discharged to the outside.

Here, the amount of the test liquid to be discharged is the first
Is determined by the taper angle of the tapered portion 48, the diameter of the boundary portion 50, the drive voltage waveform applied to the laminated piezoelectric element 32, the physical property value of the test sample, and the like.
0.3 microliter.

Thereafter, when the voltage applied to the laminated piezoelectric element 32 is gradually decreased from −E1 to E0 during the period from t4 to t5, as shown in FIG. 9E, the state of t4 <t <t5 is reached. The test sample flowing into the first tapered portion 48 is:
It gradually penetrates into the second tapered portion 49 through the boundary portion 50 and returns to the initial state at t = t5, and the test sample is filled in the second tapered portion 49 and held by surface tension. Is done.

When the discharge operation of the desired test sample is completed,
The liquid discharge head 42 is moved to the cleaning tank 22 again, and
4 to discharge the test sample remaining in the flow path 44 and to the inner surface of the flow path 44 and the flow path member 43.
Clean the outer peripheral surface of the tip. Thereafter, by repeating the above-described cleaning operation and the suction / discharge operation of the test sample,
A desired amount of the test sample is sequentially discharged into the reaction container 23.

According to the present embodiment, the discharge amount is determined by the tip diameter of the first tapered portion 48, the drive voltage waveform, the physical property value of the test sample, and the like, and the diameter of the discharge opening 46 does not influence. Therefore, the diameter of the discharge opening 46, which dries in contact with the outside air and easily causes clogging of the nozzle, can be increased without lowering the discharge accuracy, so that the occurrence of nozzle clogging can be effectively reduced with a simple configuration. Thus, a very small amount of liquid can be reliably discharged. Also, the second
The test sample in the tapered portion 49 of the first
Since the ink is ejected after being pushed back into the inside 8, the ejection flow rate is not attenuated by the test sample in the opening 39 as in the first embodiment. Therefore, the ejection operation can be performed with a low drive voltage, and the ejection efficiency can be improved.

In this embodiment, a liquid discharge head which discharges liquid by changing the volume of the flow path as shown in FIG. 6 can be used as the liquid discharge head. In the case of such a liquid discharge head, first, the piezoelectric element is driven so as to increase the flow path volume, and the test sample in the second taper portion 49 is once sucked into the first taper portion 48, and then the flow is performed. The same effect can be obtained by driving the piezoelectric element so as to reduce the passage volume and discharging the liquid from the tip of the second tapered portion 48.

In the above-described embodiment, the test sample is discharged by rapidly lowering and raising the flow path member 43 by the laminated piezoelectric element 32. For example, the test sample is rapidly lowered from the initial position and stopped. In this way, a very small amount of the test sample can be discharged, or a small amount of the test sample can be discharged by rising from the initial position and stopped, and then slowly returned to the initial position.

10 to 12 show a third embodiment of the present invention. FIG. 10 is a front view of a microarray manufacturing apparatus, FIG. 11 is a plan view, and FIG. It is a perspective view.

In the present embodiment, a microarray is manufactured by applying a DNA probe to a porous membrane filter as a substrate by applying the liquid ejection apparatus shown in the second embodiment. The same components as those in the second embodiment are denoted by the same reference numerals, and description thereof will be omitted.

This microarray manufacturing apparatus has a multi-liquid ejection head 71, an XYZ robot 72, a movable stage 75, and a washing tank 76, and a sample container 73 and a membrane filter 74 are detachably set.

The multi-liquid ejection head 71 is composed of eight liquid ejection heads 42 shown in the second embodiment arranged in the Y direction, mounted on an XYZ robot 72, and provided with a cleaning tank 76.
, And can be moved to the respective set positions of the sample container 73 and the membrane filter 74. As in the second embodiment, each of the liquid ejection heads 42 includes a pipe 25 (only one is shown, and others are omitted), a syringe piston pump 27, a solenoid valve 29, and a water supply pump 3
0, and a liquid supply and suction means having a liquid supply tank 28.

The sample container 73 has a width (Y) of 8 rows × a length (X).
It has 12 rows of 96 holes, up to 6 on the base.
It can be installed. In each hole of each sample container 73, a DNA solution containing a different type of single-stranded DNA probe labeled with a fluorescent light is stored in advance. The eight liquid discharge heads 42 in the multi liquid discharge head 71
And the arrangement pitch of the eight rows of holes of the sample container 73 are equal to each other.

The porous membrane filter 74 includes:
On this, fine droplets of the DNA solution are arranged in a two-dimensional matrix as described later, and a plurality of the DNA solutions are adsorbed and fixed on the movable stage 75 by an adsorption device (not shown). In the present embodiment, about 8 mm × 8 mm
, A maximum of 128 membrane filters 74 can be set.

Next, the operation of this embodiment will be described. First, the multi-liquid ejection head 71 is moved to the cleaning tank 76,
Eight liquid ejection heads 4 are formed in the same manner as in the second embodiment.
After cleaning the respective flow channels 44 and the outer peripheral surface of the second, a predetermined amount of air is sucked from the discharge opening 46.

Next, the multi-liquid ejection head 71 is moved to the sample container 73, and the DNA solution is ejected from each of the ejection openings 46 of the eight liquid ejection heads 42.
Is sucked into the flow path 44.

After that, the multi-liquid ejection head 71 is moved and arranged on the membrane filter 74, and the movable stage 75 and the XYZ robot 72 change the relative position between the membrane filter 74 and the multi-liquid ejection head 71 in the second embodiment. Then, the multilayer piezoelectric element 32 is driven in the same manner as described above to discharge the DNA solution. Here, the diameter of the DNA solution to be discharged is about 50 μm to 200 μm, and the diameter of the DNA solution is about 150 μm on the membrane filter 74.
Discharge at intervals of up to 400 μm.

After discharging the DNA solution onto all the set membrane filters 74, the multi-liquid discharging head 71 moves again to the cleaning tank 76, where the DNA solution remaining in the flow path 44 is discharged, and The passage 44 and the outer peripheral surface are cleaned.

Thereafter, the above-described suction / discharge and washing operations of the DNA solution are repeated, and fine droplets of different types of DNA solutions are deposited on the membrane filter 74 in the range of 150 μm to 4 μm.
Discharge is performed in a two-dimensional matrix at intervals of 00 μm.

Thus, the discharged DNA solution is permeated and held in the porous membrane filter 74,
A microarray having an A probe is manufactured.

According to the present embodiment, in addition to obtaining the same effects as in the second embodiment, a multi-liquid ejection head 71 composed of a plurality of liquid ejection heads 42 is used, and this and a membrane filter 74 are connected. A microarray can be easily and efficiently manufactured with a simple configuration in which the microarray is relatively moved in a plane perpendicular to the direction in which the DNA solution is discharged.

In this embodiment, the liquid ejection head 42 shown in the second embodiment is used. However, the liquid ejection head 20 shown in the first embodiment, or shown in FIGS. The same effect can be obtained by using such a liquid ejection head. Further, the probe sample constituting the array is not limited to a single-stranded DNA probe, for example, a ligand and a receptor for the ligand, an oligo having a specific amino acid sequence, or a polypeptide and a substance having an affinity for these peptides, An enzyme and a substrate for the enzyme,
In the case of an antigen and an antibody against the antigen, a nucleic acid having a specific base sequence, or a modified nucleic acid and a nucleic acid having a base sequence complementary to the specific base sequence of the nucleic acid or the modified nucleic acid, or a modified nucleic acid However, a microarray can be manufactured by the microarray manufacturing apparatus according to the present embodiment. These nucleic acids and modified nucleic acids include DNA, RNA, or PNA (protein nucleic acid) in which the backbone is composed of a peptide.
And so on.

Further, in the present embodiment, the porous membrane filter 74 is used as the substrate, but the present invention is not limited to this, and for example, a slide glass or a silicon substrate can be used. However, in order to prevent contamination between adjacent probes, it is preferable that the probes discharged onto the substrate be fixed at predetermined positions. In order to fix the above-described sample such as a probe on a solid-phase substrate, a functional group capable of reacting with each other may be bonded to both the probe and the substrate. Some of such methods have already been disclosed. For example, a thiol (-SH) group is bound to the end of a probe,
By treating the substrate surface so as to have a maleimide group, the thiol group of the probe supplied to the solid phase surface reacts with the maleimide group on the substrate surface to fix the probe on the substrate. Alternatively, an epoxy group may be introduced on the substrate and reacted with the amino group at the terminal of the probe.

[Brief description of the drawings]

FIG. 1 is a diagram showing a schematic configuration of a liquid ejection device according to a first embodiment of the present invention.

FIG. 2 is a sectional view of the liquid ejection head shown in FIG.

3 is a waveform diagram of a driving voltage applied to the laminated piezoelectric element shown in FIG.

FIG. 4 is a diagram for explaining an operation of discharging a test sample according to the first embodiment.

FIG. 5 is a sectional view showing a modification of the liquid ejection head used in the first embodiment.

FIG. 6 is a sectional view showing another modified example.

FIG. 7 is a sectional view illustrating a configuration of a liquid ejection head in a liquid ejection device according to a second embodiment of the present invention.

FIG. 8 is a waveform diagram of a drive voltage applied to a laminated piezoelectric element in the second embodiment.

FIG. 9 is a diagram for explaining an operation of discharging a test sample according to a second embodiment.

FIG. 10 is a front view illustrating a configuration of a microarray manufacturing apparatus according to a third embodiment of the present invention.

FIG. 11 is also a plan view.

FIG. 12 is a perspective view of the multi-liquid ejection head shown in FIG.

FIG. 13 is a view for explaining a conventional microfluidic processing apparatus.

FIG. 14 is a diagram showing a configuration of the microdispenser shown in FIG.

[Explanation of symbols]

 Reference Signs List 20 liquid discharge head 21 test sample container 22 washing tank 23 reaction container 25 pipe 27 syringe piston pump 28 liquid supply tank 29 solenoid valve 30 water feed pump 32 laminated piezoelectric element 33 gantry 34 flow path member 35 flow path 36 inlet 37 straight section 38 Tapered part 39 Opening part 40 Opening 41 Micro hole 42 Liquid discharge head 43 Flow path member 44 Flow path 45 Inlet 46 Discharge opening 47 Straight part 48 First tapered part 49 Second tapered part 50 Boundary part 71 Multi-liquid discharge Head 72 XYZ robot 73 Sample container 74 Membrane filter 75 Movable stage 76 Cleaning tank

──────────────────────────────────────────────────の Continued on the front page (51) Int. Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12N 15/09 G01N 33/566 4B029 G01N 1/00 101 37/00 102 C12M 1/00 A 31/22 121 Z33 / 566 B41J 3/04 103A 37/00 102 103Z // C12M 1/00 C12N 15/00 FF term (reference) 2C057 AF72 AG04 AG07 AG08 AG09 AG17 AG47 BA02 BA10 BA14 BF02 2G042 AA01 BD20 CB03 FB05 GA02 HA08 2G052 AA30 CA07 CA08 CA09 CA13 CA23 CA30 CA31 DA05 DA08 EA03 EA05 FC07 JA08 JA16 3E082 AA01 AA10 BB10 DD20 4B024 AA11 AA19 CA01 CA09 CA11 HA12 4B029 AA07 AA23 BB20 CC01 CC03 FA12

Claims (7)

[Claims] 1. A liquid discharging apparatus comprising: a liquid holding member having a discharge port; and a driving unit for generating a discharge pressure for discharging a small amount of liquid held by the liquid holding member from the discharge port. A liquid discharge apparatus, wherein the discharge port has an opening facing the outside atmosphere and a fine hole having a smaller sectional area than the opening. 2. The liquid discharging apparatus according to claim 1, wherein the driving unit is configured to move the liquid holding member forward and backward along a discharging direction. 3. A method of forming the tapered portion between the opening of the discharge port and the minute hole, moving the liquid holding member in the discharge direction by the driving means, and then moving the liquid holding member in the opposite direction to perform the discharge. The liquid ejection device according to claim 2, wherein the device is configured to generate pressure. 4. The liquid ejecting apparatus according to claim 1, wherein the driving unit is configured to change a volume of the liquid holding member. 5. The liquid discharging apparatus according to claim 1, wherein the driving unit includes a piezoelectric element. 6. A liquid containing a probe that can specifically bind to a target substance by the liquid discharge device using the liquid discharge device according to claim 1 on a substrate. A microarray manufacturing method, which comprises manufacturing a microarray by discharging. 7. A microarray manufacturing apparatus for manufacturing a microarray by discharging a small amount of a liquid containing a probe capable of specifically binding to a target substance onto a substrate, wherein: And a relative moving means for relatively moving the substrate and the liquid holding member in a plane perpendicular to a direction in which the liquid is ejected from the liquid holding member provided in the liquid ejection device. A microarray manufacturing apparatus, comprising: