JP2004101218A - Method and apparatus for manufacturing microarray, control method, control apparatus, and cartridge for manufacturing microarray - Google Patents

Abstract

A method and a method for manufacturing a microarray capable of manufacturing a high-density macroarray in a short time by correcting a positional deviation of a nozzle position or a landing position of a droplet by a method of software and a control method. An apparatus and a control method, a control apparatus, and a discharge container for manufacturing a microarray are provided.
A carriage on which a plurality of cartridges each having an ink jet type ejection head are mounted is moved at a constant speed in an X direction, and a table on which a substrate is set is intermittently moved in a Y direction perpendicular to the carriage movement direction. Moving the liquid and discharging the liquid in the cartridge from the discharge head onto the substrate to manufacture a microarray. In this method, beforehand, for all the discharge heads 22 of the cartridge 20, the droplet landing positions are imaged. The measurement is performed by image processing of the image data 30 and the image data, and the ejection position is corrected based on the measurement result so that the droplet is ejected to the target position.
[Selection diagram] Fig. 1

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a method and apparatus for manufacturing a microarray, which is manufactured by discharging a probe sample such as DNA, RNA, or protein onto a substrate by, for example, an inkjet method, a control method, a control apparatus, and a cartridge for manufacturing a microarray.
[0002]
[Prior art]
Genetic research institutes, medical institutions, research institutes for drug development, and the like, for DNA analysis, immunological analysis, and the like, have thousands to tens of thousands of types of DNA (deoxyribonucleic acid) and RNA on a substrate such as a slide glass. A microarray (biochip sample) in which biomolecule solutions such as (ribonucleic acid) and proteins are spotted in a matrix form is used. In order to manufacture such a microarray, for example, a method in which a liquid sample serving as a probe is attached or spotted on a substrate using a contact pin as disclosed in Patent Document 1 or a micropipette as disclosed in Patent Document 2 Has been taken.
[0003]
The contact pin method is a method in which a plurality of contact pins are moved in the X, Y, and Z directions between a liquid container and a substrate, and droplets attached to the contact pins are attached to the substrate.
The micropipette method is a method in which a micropipette containing a sample liquid is moved in the X, Y, and Z directions, pressure is applied to the sample liquid, and droplets are spotted on the substrate from the tip of the micropipette.
However, in these methods, since the number of contact pins or micropipettes is small, it is necessary to frequently move the contact pins, micropipette, or head in the X, Y, and Z directions to produce a high-density microarray. There is a problem that it takes a very long time. Also, it is very difficult to control the quantity of the droplet.
[0004]
Therefore, on the other hand, a method of producing a microarray by a capillary array by an electrophoresis method has been proposed (for example, Patent Document 3).
The electrophoresis method is a method in which a voltage is applied between a capillary and a substrate to cause the liquid in the capillary to rise by the action of an electric field and to be separated into droplets.
However, even in this method, it is necessary to move the table on which the substrate and the well or the microtiter plate are set in the X, Y, and Z directions, which is time-consuming. Also, there is a problem in increasing the density.
[0005]
On the other hand, recently, a microarray manufacturing technique of discharging a liquid serving as a probe (also referred to as a probe sample or a liquid sample) from a plurality of nozzles onto a substrate by an inkjet method has been receiving attention. This is because the inkjet method can reduce the size and density of the inkjet head, can discharge very small amounts of droplets to the target position with high accuracy, the ejection head does not contact the substrate, This is because it has features such as that any material such as glass, synthetic resin, metal, and film can be used, operation noise is low, and cost is low.
The ink-jet method is a method in which an actuator of an ink-jet head is displaced by, for example, a piezoelectric body to discharge droplets from the nozzle tip by the action of a diaphragm pump.
[0006]
As an example using such an ink-jet method, there is Patent Document 4, for example. This method uses a discharge head in which the same number of micropipette that can discharge a single liquid sample are arranged and integrated with the reservoir tank on the lower surface of a mounting plate where a plurality of reservoir tanks each containing a different type of liquid are arranged. Various types of liquid samples are ejected onto the substrate by the method.
[0007]
[Patent Document 1]
Japanese Patent Application Laid-Open No. 2000-287670 (Claims, FIG. 3)
[Patent Document 2]
US Pat. No. 5,601,980 (Claims, FIG. 1)
[Patent Document 3]
JP 2001-165904 A (Claims, FIG. 1)
[Patent Document 4]
JP 2001-186880 A (Claims, FIGS. 4 to 8)
[0008]
[Problems to be solved by the invention]
According to the inkjet method as described above, the liquid sample is discharged at a pitch of, for example, 0.2 mm × 0.2 mm by moving the discharge head and the substrate relatively in a horizontal plane and controlling the scanning of the movement. And a high-density microarray can be manufactured.
However, in order to manufacture such a high-density microarray, it is first assumed that the dimensional accuracy of the nozzle position and the nozzle interval is mechanically secured with high accuracy. However, since the ejection head of Patent Document 4 has a plurality of heads each having a single nozzle arranged side by side, as a result of accumulation of manufacturing errors and mounting errors of the heads, the nozzle position or landing of droplets is The position shifts.
In the case of a microarray, if the positional deviation between the nozzle position and the landing position of the droplet is large, adjacent liquid samples come into contact with and mix with each other and become unusable, and such a situation must be avoided. In addition, there is a limit in increasing the manufacturing accuracy in order to suppress the positional deviation of the nozzle position or the landing position of the droplet, and the cost increases. In addition, there is a limit in cost in terms of improving positioning accuracy and the like.
[0009]
Therefore, an object of the present invention is to correct the positional deviation of the nozzle position or the landing position of the liquid droplet by a method of software and control method while allowing the positional deviation due to the dimensional accuracy of the ejection head and the nozzle position. Accordingly, it is an object of the present invention to provide a method and an apparatus for manufacturing a microarray capable of manufacturing a high-density macroarray in a short time, and a control method, a control apparatus, and a cartridge for manufacturing a microarray.
[0010]
[Means for Solving the Problems]
In the method for manufacturing a microarray according to the present invention, a carriage on which a plurality of cartridges each having a droplet discharge type ejection head is mounted is moved at a constant speed in the X direction, and a table on which substrates are set is perpendicular to the movement direction X of the carriage. A method for manufacturing a microarray in which a plurality of probe samples are arranged in a matrix by intermittently moving in the Y direction and discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, for all of the ejection heads of the cartridge, the method includes a step of measuring a landing position of the droplet, and a step of correcting the ejection position to eject the droplet to a target position based on the measurement result. Features.
[0011]
According to the present invention, a plurality of cartridges having a discharge head of a droplet discharge method are mounted on a carriage, a substrate for preparing a microarray is set on a table, and the carriage and the table are scanned in the X and Y directions to form a microarray. Is what you do. Therefore, when each cartridge is mounted on the carriage, the nozzle position of the ejection head of each cartridge or the landing position of the droplet ejected from the nozzle is not always accurate.
Therefore, before actually fabricating the microarray, the displacement of the landing position of the droplet is inspected and measured. Then, based on the measurement result, the ejection position is corrected so that the droplet is ejected to the target position, so that the droplet can be ejected accurately to the target position when the microarray is manufactured.
Therefore, according to the present invention, it is possible to perform ejection with high accuracy without increasing the dimensional accuracy of the ejection head and the nozzle position of the cartridge to very high accuracy, and to manufacture a high-density microarray in a short time. it can.
In the present invention, the relationship between the carriage and the table moves relatively, and the constant speed motion and the intermittent motion may be opposite to each other.
[0012]
In the present invention, a microarray is one in which thousands to tens of thousands of types of probe samples such as DNA, RNA, and protein are arranged on a substrate such as glass. A DNA microarray (also referred to as a DNA chip) is a typical example. Further, DNA (Deoxyribonucleic Acids: deoxyribonucleic acid (genetic material)) further binds a nucleotide (nucleotide) to which a base, a sugar (deoxyribose) and a phosphate are bound, and finally forms a double helix. It has become. Here, there are four types of bases: adenine (A), guanine (G), cytosine (C), and thymine (T). Further, the DNA microarray utilizes the complementarity of DNA and is used for inspection and analysis.
RNA (ribonucleic acid: ribonucleic acid) is a nucleic acid in which ribonucleotides are polymerized, and includes ribosomal RNA, messenger RNA, transfer RNA, and the like, and is distributed not only in the nucleus but also in the whole cell. There are four types of bases: adenine (A), guanine (G), cytosine (C), and uracil (U). It has a single-stranded structure.
Protein is a general term for a high molecular weight polypeptide synthesized by peptide bonding of 20 kinds of L-amino acids as an indispensable main component of biological cells. It is a biopolymer having various molecular weights, structures and functions. is there.
[0013]
In the present invention, the step of measuring the landing position of the droplet includes the step of discharging a droplet from a nozzle of each of the discharge heads on the substrate to form a discharge test pattern; And calculating the amount of displacement in the X and Y directions of the landing position of the droplet by image processing the image data.
[0014]
In the present invention, one method for measuring the landing position of the droplet is to perform a discharge test on all the cartridges mounted on the carriage. An ejection test pattern is created (drawn), this pattern is imaged by an imaging unit, and the image data is subjected to image processing, thereby obtaining the amount of displacement in the X and Y directions of the nozzle position or the landing position of the droplet. it can.
[0015]
The imaging means is not particularly limited, but it is generally preferable to use a CCD camera.
[0016]
In the ejection position correcting step, an ejection timing correction time is obtained from the X-direction position shift amount, and an ejection timing is obtained based on the ejection timing correction time.
[0017]
In the ejection position correcting step, a table position correction value is obtained from the Y-direction positional deviation amount, and a generation time of a drive pulse for controlling a table driving motor is obtained based on the table position correction value.
[0018]
Since the carriage and the table are managed and controlled by the number of pulses for controlling the respective drive motors, for the carriage, it is necessary to determine the ejection timing correction time based on the pulse signal of the carriage drive motor from the amount of displacement in the X direction. Thus, the displacement of the ejection position in the X direction can be corrected.
In addition, for the table, since ejection is performed only when the table is stopped, a table position correction value is obtained from the amount of displacement in the Y direction, and based on the table position correction value, a driving pulse of the table drive motor for stopping the table next is used. , The displacement of the ejection position in the Y direction can be corrected.
[0019]
When each of the cartridges has a plurality of nozzles, an ejection test is performed for one or a plurality of arbitrarily selected nozzles.
[0020]
When each cartridge has a plurality of nozzles, one or a plurality of the nozzles are arbitrarily selected, or the nozzles are thinned out to perform the above-described ejection test. In the case of the ink jet system, the nozzle interval in each ejection head is generally manufactured with high accuracy, so it is not necessary to perform an ejection test on all nozzles of each cartridge. Further, by discharging different liquid samples from the plurality of nozzles, the discharge control device can be configured more compactly and inexpensively.
[0021]
Further, the microarray manufacturing apparatus according to the present invention,
A base,
A carriage that moves at a constant speed in the X direction on the base;
A table that intermittently moves in a Y direction perpendicular to the moving direction X of the carriage;
A plurality of cartridges which are detachably mounted on the carriage, have a discharge head of a droplet discharge method at a tip portion, and contain a liquid to be discharged,
One or more substrates set on the table,
Imaging means and image processing means for measuring the landing position of the droplet of each cartridge,
Discharge control means for correcting the discharge position based on the measurement result of the landing position of the droplet, and controlling the discharge timing and the table position based on the correction data,
It is characterized by having.
[0022]
With this configuration, the microarray manufacturing method of the present invention described above can be efficiently performed, and a high-density microarray can be manufactured with high production efficiency.
[0023]
Further, in the microarray manufacturing apparatus of the present invention, the carriage is electrically connected to the cartridge when the cartridge is mounted.
[0024]
With this configuration, each cartridge can be detachably mounted on the carriage, and is electrically connected at the time of mounting, so that the droplets are ejected from the nozzles of the ejection head of each cartridge by a droplet ejection method. Is discharged onto a substrate to manufacture a microarray. Further, replacement of the cartridge is easy.
[0025]
The cartridge is configured such that the ejection head having one or a plurality of nozzles and a tank for containing a liquid to be ejected are integrally formed, or a plurality of nozzles and a plurality of reservoir tanks are independently provided. This is a configuration having a discharge head that communicates.
[0026]
The cartridge used in the present invention is configured as described above, and the tank of each cartridge contains a biomolecule solution for generating a probe sample. The liquids can all be of the same type or different types. Moreover, the cartridge is easy to handle and replace.
[0027]
Further, in the control method for manufacturing a microarray according to the present invention, a carriage on which a plurality of cartridges each having a droplet discharge type ejection head is mounted is moved at a constant speed in the X direction, and a table on which a substrate is set is moved by the carriage. In the manufacture of a microarray in which a plurality of probe samples are arranged in a matrix by intermittently moving in the Y direction perpendicular to the direction X and discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, performing a discharge test on the total number of discharge heads of the cartridge, and correcting the discharge position so as to discharge the droplet to the target position based on the measurement result of the landing position of the droplet in the discharge test pattern,
Controlling a discharge timing and a table position based on the correction data of the discharge position,
It is characterized by including.
[0028]
As described above, the control method of the present invention performs an ejection test on all the ejection heads of the cartridge in advance, and corrects the ejection position based on the measurement result of the landing position of the droplet in the ejection test pattern. Since the ejection timing and the table position are controlled based on the ejection position correction data, the droplet can be ejected to the target position almost accurately.
[0029]
In this case, with respect to the movement direction X (constant speed movement direction) of the carriage, the control step obtains the number of accumulated pulses from the mechanical origin for controlling the carriage drive motor and the displacement amount of the landing position of the droplet in the X direction. The ejection timing is controlled by carriage position control data including the ejection timing correction time.
Here, the mechanical origin is the coordinate origin defined by the microarray manufacturing apparatus, and generally indicates the home position of the carriage.
[0030]
Further, with respect to the table movement direction Y (intermittent movement direction), the control step controls the table drive motor by table position control data including a table position correction value obtained from the Y direction displacement amount of the droplet landing position. I do.
[0031]
Further, the control device for manufacturing a microarray according to the present invention moves a carriage on which a plurality of cartridges each having a droplet discharge type discharge head is mounted at a constant speed in the X direction, and moves a table on which a substrate is set to a position where the carriage moves. In the manufacture of a microarray in which a plurality of probe samples are arranged in a matrix by intermittently moving in the Y direction perpendicular to the direction X and discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, an imaging unit and an image processing unit for performing an ejection test on all the ejection heads of the cartridge, and measuring a landing position of a droplet in an ejection test pattern,
Position shift calculating means for calculating the amount of shift in the X and Y directions of the landing position of the droplet;
Discharge timing correction means for calculating a discharge timing correction time from the amount of displacement of the landing position of the droplet in the X direction,
Table position correction means for calculating a table position correction value from the amount of displacement of the landing position of the droplet in the Y direction,
Discharge control means for controlling the discharge timing and the table position based on the correction data of the discharge position,
It is characterized by having.
[0032]
The discharge control unit generates carriage position control data including a cumulative number of pulses from a mechanical origin for controlling a carriage drive motor and a discharge timing correction time obtained from an X-direction displacement amount of a landing position of the droplet. To control the ejection timing.
[0033]
The ejection control means controls the table drive motor by generating table position control data including a table position correction value obtained from the Y direction displacement of the landing position of the droplet.
[0034]
Further, a cartridge for producing a microarray according to the present invention is a cartridge used in the method for producing a microarray according to any one of claims 1 to 6 or the apparatus for producing a microarray according to any one of claims 7 to 10. So,
The cartridge is a cartridge in which a discharge head of a droplet discharge method having one or a plurality of nozzles and a tank for storing a liquid to be discharged are integrally formed.
[0035]
The cartridge is configured such that the ejection head and the carriage are electrically connected when the cartridge is mounted on the carriage.
[0036]
With the cartridge of the present invention configured as described above, the droplets can be ejected by the ink jet method and can be freely attached to and detached from the carriage, so that the handling and replacement are easy. Further, since the ejection position is corrected as described above, it is not necessary to increase the dimensional accuracy of the cartridge, and the cost can be reduced.
[0037]
BEST MODE FOR CARRYING OUT THE INVENTION
Embodiment 1 FIG.
FIG. 1 is a perspective view showing an outline of a microarray manufacturing apparatus according to Embodiment 1 of the present invention, FIG. 2 is a schematic sectional view of a carriage and a cartridge mounted thereon, FIG. 3 is a plan view of the carriage, and FIGS. Is an enlarged perspective view, a sectional view, and a bottom view of the cartridge.
The microarray manufacturing apparatus 10 includes, on a base 11, a carriage 12 that moves at a constant speed in the X direction in a horizontal plane, a cartridge type cartridge 20 that is mounted on the carriage 12 side by side, and a cartridge perpendicular to the X axis in the horizontal plane. A table 14 intermittently moved at a predetermined feed pitch in the Y direction, a substrate 100 made of a slide glass, resin, or the like, which is set (placed) on the table 14, and discharged onto the substrate 100. Imaging means 30 for detecting the landing position of the ejected droplets; and further, drive control means 40 for the X-axis (carriage) and Y-axis (table) and ejection control means (not shown) based on image signals from the imaging means 30. Zu). The ejection control means is generally composed of a PC (personal computer) and a dedicated circuit.
In this example, the carriage 12 is moved at a constant speed and the table 14 is moved intermittently. However, the carriage 12 may be moved at an intermittent speed and the table 14 may be moved at a constant speed.
[0038]
On the carriage 12, a plurality of cartridges 20 are removably mounted side by side. Therefore, the carriage 12 is also a cartridge holder. The cartridges 20 are arranged in 4 rows × 6 columns at a pitch of, for example, 10 mm × 10 mm.
As shown in FIGS. 5 and 6, the cartridge 20 includes a case 21 for a cover, a discharge head 22 attached to a front end (lower end) of the case 21 and discharging droplets by, for example, an inkjet method. A liquid tank 28 made of rubber or the like containing a liquid 27 for ejection (for example, a biomolecule solution) is integrally formed. Further, a wiring portion 25 for driving the actuator of the discharge head 22, contacts 26 a and 26 b of the wiring portion 25 provided on the lower surface of the case 21, and a liquid supply port 24 for supplying a discharge liquid to the liquid tank 28 are provided. I have. The hard case 21 itself may be used as a liquid tank by omitting the liquid tank 28 made of rubber or the like.
[0039]
The bottom of the carriage 12 has a through-hole 121 into which the ejection head 22 of the cartridge 20 is closely fitted, and the electrodes 122a and 122b which are in contact with the contacts 26a and 26b and are electrically connected when the cartridge 20 is mounted. The wiring to the electrodes 122a and 122b of each cartridge 20 is connected to the ejection control means via a connector 124 provided on the side surface of the carriage 12.
[0040]
The carriage 12 starts moving from a home position that is the mechanical origin O, and is controlled so as to maintain a constant speed at least within the range of the ejection area on the table 14.
Although the X-axis driving mechanism of the carriage 12 is not shown, it can be implemented by using a known timing belt mechanism, a ball screw mechanism, or the like, and coupling the carriage 12 to the mechanism to drive a pulse motor. In FIG. 1, such an X-axis driving mechanism is built in the X-axis duct 15, and further, an elongated hole 16 of the carriage 12 is provided in the X-axis duct 15. An encoder is attached to the pulse motor that drives the carriage 12, for example.
[0041]
On the table 14, one or a plurality of substrates 100 for microarray fabrication are set (placed). When performing the ejection test, one substrate is selected from the actually used microarray manufacturing substrates 100 and the ejection test is performed. Of course, a test substrate may be used separately.
The table 14 is intermittently fed in the Y direction by a Y-axis drive mechanism (not shown). This Y-axis drive mechanism has the same configuration as the X-axis drive mechanism, is built in the Y-axis duct 17 of FIG. 1, and further has an elongated hole 18 of the table 14 in the Y-axis duct 17. .
[0042]
The imaging means 30 uses, for example, a CCD camera. In this case, as the image sensor, a two-dimensional sensor is more appropriate than a one-dimensional sensor. For example, the imaging unit 30 is attached to the side wall in the Y direction and installed so as to capture an image of a droplet pattern (ejection test pattern) ejected on the substrate 100. This image signal is sent to an image processing unit in the ejection control means.
[0043]
Next, the nozzle structure and operation of the ejection head 22 of one cartridge 20 will be described with reference to FIG.
The ejection head 22 has one or a plurality of nozzles (ejection ports) 220. The nozzles 220 are formed at equal intervals, for example, at 0.5 mm intervals. The discharge head 22 is formed by joining an upper glass substrate 221, an intermediate Si substrate 222 in FIG. 7 and a lower glass substrate 223 in FIG. 7 by anodic bonding with each other. A nozzle 220, a discharge chamber 224, and an orifice 225, which constitute a flow path portion by etching, are formed in advance at 222. The bottom of the discharge chamber 224 serves as a vibration plate 227 for discharging droplets.
[0044]
A concave portion 228 is formed in the lower glass substrate 223, and an actuator electrode 229 is formed in the concave portion 228 so as to face the diaphragm 227 with a predetermined gap. Then, by driving an oscillation circuit (not shown) connected between the actuator electrode 229 and the Si substrate 222, an electrostatic attraction is generated to displace the vibration plate 227, thereby increasing the volume of the discharge chamber 224. Thus, the liquid 23 is sucked, and the vibration plate 227 is restored, whereby the droplet is discharged from the nozzle 220. As a driving method of the vibration plate 227, a piezoelectric actuator may be used in addition to the above-described electrostatic actuator.
[0045]
Next, a control system according to the present embodiment will be described. FIG. 8 shows an example of an ejection test pattern for detecting a positional shift between a nozzle position or a droplet landing position. FIG. 9 is a control block diagram for correcting the discharge position, FIG. 10 is a flowchart for correcting the discharge position, and FIG. 11 is a timing chart showing the discharge timing after the discharge position correction, the table motor drive timing, and the like. .
9, reference numeral 31 denotes a CCD camera of the image pickup means 30, 51 denotes an image processing unit, 52 denotes a displacement calculating unit, 41 denotes an X-axis encoder, 53 denotes an ejection timing correcting unit, 54 denotes a table position correcting unit, and 42 denotes a head drive. A controller 43 is a table drive motor controller, 22 is an ejection head of each cartridge 20, and 44 is a table drive motor.
[0046]
The cartridges 20 are arranged on the carriage 12 in M rows × N columns. At this time, each cartridge 20 is mounted with its ejection head 22 closely fitted to the through-hole 121 of the carriage 12. Although the dimensions of the through-holes 121 and the pitch intervals thereof are manufactured with a high degree of precision by machining, there is a limit to this, so that the nozzle positions are misaligned as a whole during mounting. As a result, the landing position of the droplet also deviates from the predetermined target position. This is a major problem in realizing high density in the manufacture of microarrays.
[0047]
Therefore, first, an ejection test is performed in advance, the displacement of each nozzle position is measured, and based on the measurement result, the ejection position is corrected so that the droplet is ejected to the target position, and then the actual microarray Is going to manufacture. Note that it is sufficient to perform the ejection test once as long as the cartridge 20 is used.
[0048]
Referring to FIGS. 9 and 10, first, all necessary cartridges 20 are set on the carriage 12 (S1 in FIG. 10), and droplets are ejected from the nozzles 220 of each cartridge 20 onto the substrate 100. As a result, an ejection test pattern 300 for detecting the positional deviation, which represents the positional deviation of the nozzle position, as shown in FIG. 8, is created (drawn) (S2 in FIG. 10). FIG. 8 shows the landing position of the reference cartridge and the actual ejection position (or the actual landing position) based on the displacement of the nozzle 220 of each cartridge 20.
[0049]
In order to form the ejection test pattern 300, the carriage 12 is scanned at a constant speed in the X direction and the table 14 is scanned in the Y direction on the substrate 100 of the table 14, and the ejection signal is generated at predetermined time intervals. The ejection causes the droplets to be ejected onto the substrate 100 from the nozzles 220 of the ejection heads 22 of the reference cartridge and each cartridge 20, whereby the ejection test pattern 300 can be created (drawn). In this case, as shown in FIG. 8, all the cartridges are discharged, for example, three times for each cartridge, and the average value of the amount of displacement is determined. The number of ejections is not limited and may be one at a time.
[0050]
The ejection test pattern 300 thus obtained is imaged by the CCD camera 31 of the imaging means 30. The image data is input to the image processing unit 51 of the ejection control unit, and after being binarized by the image processing unit 51, the landing position (x, y coordinate values) of the droplet is measured. Then, the displacement calculating unit 52 calculates the displacement (average value) in the X direction and the Y direction of each impact position with respect to the target ejection position for each cartridge 20 (S3 in FIG. 10).
The calculated X-direction displacement amount (average value) is input to the ejection timing correction unit 53, where the correction time dt of the ejection timing is calculated by the following equation (S4 in FIG. 10).
[0051]
(Equation 1)
dt = Σ (xn−x0) / N · v (n = 1, 2, 3,... N)
Where: v: carriage speed (constant)
N: Number of test ejections
[0052]
The amount of displacement (average value) in the Y direction from the displacement calculator 52 is input to the table position corrector 54, where the table position correction value dy is calculated by the following equation (S4 in FIG. 10).
[0053]
(Equation 2)
dy = Σ (yn−y0) / N (n = 1, 2, 3,... N)
In the calculation processing of the correction data in S4 of FIG. 10, the carriage position correction data and the table position correction value dy obtained from Expression (1) and the table position correction value dy obtained from Expression (2) are respectively used. Table position correction data is calculated.
[0054]
The ejection timing correction unit 53 converts the carriage position correction data based on the ejection timing correction time dt into a pulse signal of an X-axis encoder (an encoder provided on the carriage drive motor, which is composed of an encoder scale and an encoder sensor) 41. Is converted to time data T1 (see FIG. 11) based on the reference, and is further converted to (T0 + T1) carriage position control data as shown in the timing chart of FIG. 11 (S5 in FIG. 10).
Here, T0 is the time from the home position (mechanical origin O) of the carriage 12 to the target ejection position, and is managed by the number of pulses (cumulative number of pulses) of the X-axis encoder 41. Further, T1 is a time until the ejection based on the pulse signal of the X-axis encoder 41, and is a value corrected based on the ejection timing correction time dt obtained from the information of the preliminary ejection test drawing as described above.
[0055]
Further, the correction data based on the table position correction value dy calculated by the table position correction unit 54 is converted into table position control data including the time T3 as shown in FIG. 11 (S5 in FIG. 10). Here, T3 is a time (table movement start time) until the table 14 starts to move based on the corrected ejection signal, and the table 14 moves to the next ejection position. No ejection is performed while the table 14 is moving. Discharge is performed while the table is stopped. T4 is a pulse generation time for moving the table, and is a value corrected based on the table position correction value dy obtained from the information of the previous ejection test drawing. The movement of the table 14 is managed by the number of pulses including acceleration and deceleration. In FIG. 11, T2 is a pulse width for driving and ejecting the actuator of the ejection head 22.
[0056]
The carriage position control data is sent to the head drive control unit 42, and the head drive control unit 42 issues an ejection signal to the ejection head 22 of the cartridge 20 at the ejection timing corrected as described above. At this time, an ejection pulse having a pulse width of T2 is applied to the actuator of the ejection head 22.
Further, the table drive motor control unit 43 controls the table drive motor 44 based on table position control data including a table movement start time T3 based on the ejection signal.
[0057]
As described above, after the ejection positions of all the cartridges 20 are corrected to be the target ejection positions, the ejection head 22 starts ejection (S6 in FIG. 10). Even if the nozzle position is misaligned, it is possible to discharge the ink to the target position almost exactly.
Since each cartridge 20 can discharge droplets on the order of several pl (picoliter) by an ink-jet method, it is possible to manufacture a high-density microarray, for example, at a pitch of 0.2 mm × 0.2 mm. .
In addition, since it is not necessary to stop at each step, high-speed spotting is possible, and the production efficiency of the microarray is high.
Further, since high dimensional accuracy is not required for the cartridge 20, it is advantageous in terms of cost.
The cartridge 20 only needs to insert the ejection head 22 into the through-hole 121 of the carriage 12, so that the cartridge 20 can be easily attached and detached when replacing the cartridge, and does not require complicated adjustment. Therefore, if the position, the shape, and the like of the through-hole 121 are devised so as not to mistake the direction of the cartridge, a special positioning means is unnecessary.
Since the ejection heads 22 are constituted by independent cartridges 20, there is no mixing of liquids.
[0058]
Embodiment 2 FIG.
When the ejection head 22 of each cartridge 20 has a plurality of nozzles 220 as shown in the cross-sectional view of FIG. 5, one or a plurality of nozzles 220 are arbitrarily selected from among them, and for example, in FIG. , And the first, third, and sixth nozzles are selected, and the ejection positions are corrected for these specified nozzles 220 and for all cartridges 20 as described in the first embodiment. Of course, the nozzle position may be corrected for all the nozzles 220 of each cartridge 20, but it takes time and the nozzle pitch interval is usually manufactured with high precision. If the shift is a problem, the handling described above does not cause a significant problem.
Further, a method of thinning out nozzles at regular intervals may be adopted.
[0059]
Embodiment 3 FIG.
FIG. 12 is a schematic configuration diagram of a multi-reservoir type ejection head mounted on a cartridge. The cartridge 20 described in the first embodiment is configured to mainly discharge one type of biomolecule container. On the other hand, in this embodiment, a plurality of types of biomolecule solutions can be discharged by each cartridge or one cartridge.
Therefore, the multi-reservoir head 22A attached to the front end (lower end) of each cartridge 20 has a configuration in which the plurality of nozzles 220 communicate with the plurality of reservoir tanks 226 via independent flow paths 230. . Further, as shown in FIG. 7, the discharge section including the nozzle 220 includes a nozzle, a discharge chamber, a diaphragm, an actuator electrode, and the like.
The correction of the ejection position with respect to the multi-reservoir head 22A may be performed for one or a plurality of arbitrarily selected nozzles 220, as described in the second embodiment.
Therefore, if the cartridge 20 having the multi-reservoir head 22A configured as described above and having the ejection position corrected is used, thousands to tens of thousands of types of microarray groups can be manufactured in a short time.
[0060]
【The invention's effect】
As described above, according to the present invention, the landing positions of droplets are measured for all of the droplet discharge type cartridges mounted on the carriage, and the discharge positions are corrected based on the measurement results. Even if the dimensional accuracy of the microarray is not high, there is no need to increase the dimensional accuracy, and there is an effect that a high-density microarray can be economically manufactured and the production efficiency can be improved.
[Brief description of the drawings]
FIG. 1 is a schematic perspective view of a microarray manufacturing apparatus according to an embodiment of the present invention.
FIG. 2 is a schematic sectional view of the carriage of FIG. 1 and a cartridge mounted thereon.
FIG. 3 is a plan view of the carriage of FIG. 1;
FIG. 4 is an enlarged perspective view of the cartridge.
FIG. 5 is a sectional view of the cartridge.
FIG. 6 is a bottom view of the cartridge.
FIG. 7 is a sectional view of a discharge head of the cartridge.
FIG. 8 is a diagram illustrating an example of an ejection test pattern for detecting a positional shift of a nozzle position or a landing position of a droplet.
FIG. 9 is a control block diagram for correcting a discharge position.
FIG. 10 is a flowchart for correcting a discharge position.
FIG. 11 is a timing chart of discharge position control.
FIG. 12 is a schematic configuration diagram of a multi-reservoir type ejection head.
[Explanation of symbols]
10: Microarray manufacturing apparatus, 11: Base, 12: Carriage, 14: Table, 15: X-axis duct, 16: Guide slot, 17: Y-axis duct, 18: Guide slot, 20: Cartridge, 21 : Case, 22: discharge head, 23: liquid, 24: liquid supply port, 25: wiring section, 26a, 26b: contact point, 27: liquid, 28: liquid tank, 30: imaging means, 31: CCD camera, 40: Drive control means, 41: X-axis encoder, 42: Head drive control section, 43: Table drive motor control section, 44: Table drive motor, 51: Image processing section, 52: Position shift correction section, 53: Ejection timing correction section , 54: table displacement correction unit, 100: substrate, 121: through hole, 122a, 122b: electrode, 124: connector, 220: nozzle, 221: glass substrate, 222: Si substrate, 223: glass substrate, 224: discharge chamber, 225: orifice, 226: reservoir tank, 227: diaphragm, 228: recess, 229: actuator electrode, 230: flow path, 300: discharge test pattern

Claims (18)

A carriage equipped with a plurality of cartridges having a discharge head of a droplet discharge method is moved at a constant speed in the X direction, and a table on which substrates are set is intermittently moved in a Y direction perpendicular to the movement direction X of the carriage. A method for producing a microarray in which a large number of probe samples are arranged in a matrix by discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, for all of the ejection heads of the cartridge, the method includes a step of measuring a landing position of the droplet, and a step of correcting the ejection position to eject the droplet to a target position based on the measurement result. A method for producing a microarray. The step of measuring the landing position of the droplet includes a step of forming a discharge test pattern by discharging a droplet from the nozzle of each of the discharge heads on the substrate, and an image of the discharge test pattern by an imaging unit. 2. The method for manufacturing a microarray according to claim 1, further comprising the step of: performing image processing on the data to determine a displacement amount of the landing position of the droplet in the X and Y directions. 3. The method according to claim 2, wherein a CCD camera is used as the imaging unit. 4. The discharge position correcting step according to claim 1, wherein a discharge timing correction time is obtained from the X-direction displacement amount, and a discharge timing is obtained based on the discharge timing correction time. A method for producing a microarray. The discharge position correction step includes obtaining a table position correction value from the Y-direction position shift amount, and obtaining a drive pulse generation time for controlling a table drive motor based on the table position correction value. Item 5. The method for producing a microarray according to any one of Items 1 to 4. 6. The method according to claim 1, wherein when the cartridge has a plurality of nozzles, an ejection test is performed on one or a plurality of nozzles arbitrarily selected. A base,
A carriage that moves at a constant speed in the X direction on the base;
A table that intermittently moves in a Y direction perpendicular to the moving direction X of the carriage;
A plurality of cartridges which are detachably mounted on the carriage, have a discharge head of a droplet discharge method at a tip portion, and contain a liquid to be discharged,
One or more substrates set on the table,
Imaging means and image processing means for measuring the landing position of the droplet of each cartridge,
Discharge control means for correcting the discharge position based on the measurement result of the landing position of the droplet, and controlling the discharge timing and the table position based on the correction data,
An apparatus for manufacturing a microarray, comprising: 8. The microarray manufacturing apparatus according to claim 7, wherein the carriage is configured to be electrically connected to the cartridge when the cartridge is mounted. 9. The microarray manufacturing apparatus according to claim 7, wherein the cartridge integrally includes the discharge head having one or a plurality of nozzles and a tank for storing a liquid to be discharged. 10. The microarray manufacturing apparatus according to claim 7, wherein the cartridge has a discharge head in which a plurality of nozzles and a plurality of reservoir tanks communicate with each other independently. A carriage equipped with a plurality of cartridges having a discharge head of a droplet discharge method is moved at a constant speed in the X direction, and a table on which substrates are set is intermittently moved in a Y direction perpendicular to the movement direction X of the carriage. In the manufacture of a microarray in which a large number of probe samples are arranged in a matrix by discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, performing a discharge test on the total number of discharge heads of the cartridge, and correcting the discharge position so as to discharge the droplet to the target position based on the measurement result of the landing position of the droplet in the discharge test pattern,
Controlling a discharge timing and a table position based on the correction data of the discharge position,
A control method for manufacturing a microarray, comprising: The discharge timing control step is based on carriage position control data including a cumulative number of pulses from a mechanical origin for controlling the carriage drive motor and a discharge timing correction time obtained from an amount of displacement of the landing position of the droplet in the X direction. The control method for manufacturing a microarray according to claim 11, wherein the ejection timing is controlled. 12. The table position control data according to claim 11, wherein the table position control step controls the table drive motor by table position control data including a table position correction value obtained from a Y direction displacement amount of the droplet landing position. Control method for microarray manufacturing. A carriage equipped with a plurality of cartridges having a discharge head of a droplet discharge method is moved at a constant speed in the X direction, and a table on which substrates are set is intermittently moved in a Y direction perpendicular to the movement direction X of the carriage. In the manufacture of a microarray in which a large number of probe samples are arranged in a matrix by discharging the liquid in the cartridge from the discharge head onto the substrate,
In advance, an imaging unit and an image processing unit for performing an ejection test on all the ejection heads of the cartridge, and measuring a landing position of a droplet in an ejection test pattern,
Position shift calculating means for calculating the amount of shift in the X and Y directions of the landing position of the droplet;
Discharge timing correction means for calculating a discharge timing correction time from the amount of displacement of the landing position of the droplet in the X direction,
Table position correction means for calculating a table position correction value from the amount of displacement of the landing position of the droplet in the Y direction,
Discharge control means for controlling the discharge timing and the table position based on the correction data of the discharge position,
A control device for manufacturing a microarray, comprising: The discharge control means generates carriage position control data including a cumulative number of pulses from a mechanical origin for controlling a carriage drive motor and a discharge timing correction time obtained from an X-direction displacement amount of a landing position of the droplet. 15. The control device for manufacturing a microarray according to claim 14, wherein the ejection timing is controlled by using the control device. 15. The apparatus according to claim 14, wherein the ejection control unit controls the table drive motor by generating table position control data including a table position correction value obtained from a displacement amount of the landing position of the droplet in the Y direction. For microarray production. A cartridge used in the method for manufacturing a microarray according to any one of claims 1 to 6 or the apparatus for manufacturing a microarray according to any one of claims 7 to 10.
A cartridge for manufacturing a microarray, wherein the cartridge is a cartridge in which a droplet discharge type discharge head having one or a plurality of nozzles and a tank for storing a liquid to be discharged are integrally configured. 18. The cartridge for manufacturing a microarray according to claim 17, wherein the cartridge is configured to electrically connect the ejection head and the carriage when the cartridge is mounted on the carriage.

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