JP2007278986A - Apparatus and method for dispensation and apparatus and method for manufacturing microarray - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a dispensing technique capable of reducing variations in spot size. <P>SOLUTION: A dispensing apparatus 1 is provided with a dispensing unit 100 and a first moving means 202. The dispensing unit 100 includes a holding part 104 for holding a liquid containing a biological sample; a pressing means 106 for pressing the held liquid; and a dispensing needle 102 capable of discharging droplets and can form a droplet 20 at the tip of the dispensing needle 102, by pressing the liquid by the pressing means 106. The first moving means 202 moves the location of the tip of the dispensing needle 102, approximately in a vertical direction and fixes it, in such a way that the distance between the tip of the dispensing needle 102 and an object surface of discharge of an object 10 to which a scheduled droplet 20 is to be discharged approximately in the vertical direction to the object surface of discharge, is approximately equal to the length of the scheduled droplet in its dropping direction formed at the tip of the dispensing needle 102. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a dispensing technique. Specifically, the present invention relates to a technique for dispensing a minute amount of a liquid containing a biological sample used for creating a microarray.

  Biochips (microarrays) such as DNA chips, RNA chips, protein chips, and sugar chain chips align hundreds to tens of thousands of biological materials and chemical substances on a substrate such as a glass slide of several square centimeters at high density. , Fixed. Biochips are used to search for substances that interact with substances on the chip and to obtain information from signal patterns of interacting substances. Biochips are expected in various fields such as therapeutic drug development tools, disease diagnosis, and health monitoring because they can process and analyze large amounts of information at once.

  The method for producing a biochip can be roughly classified into an optical lithographic method, a surface adhesion method, and an ink jet method.

  Among the dispensing methods, the surface adhesion method is a method in which the needle tip of a dispensing needle is immersed in a liquid containing a biological sample such as a DNA fragment and then the liquid is spotted by contacting the needle tip on a substrate. It is. The dispensing needle can be reused by washing with a washing solution or the like, and various types of biological samples can be spotted using the same dispensing needle. This method is advantageous in that the cost is low and it is easy to create a unique arrangement pattern. However, on the other hand, when the tip of the needle is first immersed in the liquid, the spot size may vary between the first spot and the subsequent spot due to the extra liquid adhering around the tip of the dispensing needle. Alternatively, the spot size may vary depending on contact conditions such as contact time and contact angle between the needle tip and the substrate.

Under such circumstances, in order to solve the above problem, Japanese Patent No. 3436741 (Patent Document 1) discloses that after the dispensing needle is immersed in the liquid, excess liquid adhering to the needle tip is discarded. The method of removing by this is disclosed. Moreover, in JP 2004-325329 A (Patent Document 2), the distance between the dispensing pin and the dispensing object is kept constant by the positioning pin, the tip of the dispensing pin is not brought into contact with the surface of the dispensing object, A method is disclosed in which a droplet containing protein held at the tip of a dispensing pin is brought into contact with a dispensing object and dispensed in a non-contact manner by the surface tension of the droplet.
Japanese Patent No. 3436741 JP 2004-325329 A

  However, in the method of Patent Document 1, since the dispensing needle directly contacts the substrate, there is a possibility that variation due to contact may occur. Further, in the method of Patent Document 2, since the liquid is collected by immersing the tip of the dispensing pin in the liquid, the spot size varies in the first and subsequent spots due to the excess liquid attached around the tip of the dispensing pin. There is a fear. In addition, since the dispensing pin is moved in a state where the droplet is held, the droplet may drop at an unplanned place or timing.

  Accordingly, an object of the present invention is to provide a dispensing technique that can further reduce the variation in spot size. Another object of the present invention is to provide a dispensing technique capable of avoiding contamination that may be caused by dropping of a droplet at an unplanned place or timing.

  In order to solve the above-mentioned problems, the present invention comprises a holding unit that holds a liquid containing a biological sample, a pressing unit that presses the held liquid, and a dispensing needle that can discharge liquid droplets. The dispensing target between the dispensing unit capable of forming a droplet at the tip of the dispensing needle by pressing the liquid by the pressing means, and the ejection target surface of the object to be ejected The tip position of the dispensing needle is set in the substantially vertical direction so that the distance in the substantially vertical direction with respect to the surface is substantially the same as the length in the drop direction of the droplet to be formed at the tip of the dispensing needle. And a first moving means for moving and fixing the dispensing device.

  According to this configuration, since the dispensing unit is provided with the pressing means, it is possible to form a droplet at the spot forming position of the discharge target object, and the droplet falls at an unscheduled place or timing. It is possible to avoid contamination that may occur due to this. In addition, since the tip position of the dispensing needle can be moved and fixed to a position that is approximately the same distance as the length of the droplet to be formed, the tip position of the dispensing needle can be fixed. And the liquid droplets are formed until they come into contact with the surface to be ejected, so that the tip of the needle is separated from the surface to be ejected by the surface tension of the liquid droplets (without contact). It becomes possible to note. Therefore, it is possible to avoid variation in spot size due to the dispensing needle coming into contact with the discharge target object. Further, in the present invention, when supplying the liquid containing the biological sample to the dispensing needle, it is not necessary to immerse the dispensing needle in the liquid, and there is no possibility of excess liquid adhering around the tip of the dispensing needle. Therefore, it is possible to avoid variation in spot size due to excess liquid adhering to the tip of the dispensing needle.

  Preferably, there is provided second moving means for moving the dispensing unit relatively to the spot forming position on the discharge target object relatively horizontally with the discharge target surface of the discharge target object. According to this, the dispensing unit can be moved three-dimensionally to a desired position on the ejection target surface.

  Preferably, the tip of the dispensing needle is subjected to a liquid repellent treatment. According to this, it is possible to promote the transfer of the liquid droplet to the target object when the liquid droplet comes into contact with the target object.

  Preferably, a lyophilic process is performed on the discharge target object. According to this, it becomes possible to promote the transfer of droplets to the discharge target object.

  Preferably, the pressing means is means for pressing the liquid by moving a plunger provided in the dispensing unit by a feed screw mechanism. According to this, since a very small amount of liquid can be adjusted, the size of the droplet at the tip of the dispensing needle can be easily adjusted to a desired size.

  Preferably, the feed screw mechanism is driven by an actuator. According to this, it is possible to automatically control the droplet size.

  Preferably, the dispensing apparatus of the present invention stores the relationship between the input means for inputting a desired spot size, the distance between the tip of the dispensing needle and the discharge target object, and the spot size in a readable manner. A first drive signal is transmitted to the first moving means so as to move the distance read from the storage means in response to the input of the spot size to the storage means and the input means. A movement control means for controlling one movement means; a pressure control means for transmitting a second drive signal to the pressure means after the tip position of the dispensing needle has been moved and fixed to the distance; Detecting means for detecting a positional relationship between a droplet formed at the tip of the needle and a discharge target surface of the discharge target body, wherein the pressure control means further detects the liquid detected by the detection means. Droplet and target surface Transmitting a stop signal to stop the drive to the pressing means in accordance with the positional relationship.

  According to this, by inputting a desired spot size, it becomes possible to automatically form droplets having a size corresponding to the distance between the tip of the dispensing needle and the surface of the object to be ejected, improving work efficiency. Can be achieved. Here, the positional relationship between the droplet and the ejection target surface includes, for example, the distance between the tip of the droplet and the ejection target surface.

  Preferably, the detection means is configured to observe a liquid formed on the tip of the dispensing needle, an observation means for observing the discharge target surface of the discharge target object, and a light image observed by the observation means. And determining means for calculating a distance between the droplet tip and the ejection target surface, comparing the distance with a predetermined value, and determining whether or not the distance matches the predetermined value. According to this, it becomes possible to control the driving of the pressing means with high accuracy. Here, examples of the observation means include optical observation means such as a CCD (Charge Coupled Devices) camera.

  Further, the detection means is constituted by a laser light source and a light receiving sensor instead of the detection means using the observation means, and from the change in received light intensity when the droplet ejected from the tip of the dispensing needle crosses the laser beam. A mechanism may be used to detect the droplet position. According to this, it is possible to detect the positional relationship between the droplet and the ejection target surface with a simpler configuration.

  The pressing control unit may transmit the second driving signal after a predetermined time after the first driving signal is stopped. According to this, it is possible to continuously move the dispensing unit and form droplets with a simple configuration.

  Another aspect of the present invention provides a microarray manufacturing apparatus provided with the above-described dispensing apparatus. According to this, since the dispensing device is provided, contamination during the production of the microarray can be avoided, and variation in spot size can be reduced, so that the accuracy and yield of the microarray can be improved. Can do.

  Another aspect of the present invention is a dispensing method in which a liquid containing a biological sample is dispensed to an object to be ejected with a dispensing needle, the tip of the dispensing needle and the ejection target surface of the object to be ejected. The tip position of the dispensing needle is moved so that the distance in the direction substantially perpendicular to the surface to be ejected coincides with the length of the liquid drop to be formed at the tip of the dispensing needle. The liquid held by the dispensing needle is exuded to the tip of the dispensing needle by pressing with the pressing means, and the droplet formed at the tip of the dispensing needle is gradually grown. A dispensing method is provided in which pressing the liquid droplet tip is brought into contact with the discharge target body and the pressing is stopped almost simultaneously, and the liquid containing the biological sample is moved to the discharge target body.

  According to this, after the tip of the dispensing needle is moved and fixed to a position that coincides with the length in the drop direction of the droplet to be formed, the liquid held by the dispensing needle is pressed by the pressing means. Therefore, it is possible to avoid contamination by dropping the liquid droplets at an unplanned place or timing. Further, when the tip of the dispensing needle is not brought into contact with the object to be ejected (without contact) and the tip of the liquid droplet is brought into contact with the object to be ejected, the liquid is applied to the object to be ejected by the surface tension of the liquid droplet. Since the droplets are transferred, it is possible to avoid variations in spot size due to the dispensing needles coming into contact with the discharge target object. Further, in the present invention, when supplying the liquid containing the biological sample to the dispensing needle, it is not necessary to immerse the dispensing needle in the liquid, and there is no possibility of excess liquid adhering around the tip of the dispensing needle. Therefore, it is possible to avoid variation in spot size due to excess liquid adhering to the tip of the dispensing needle.

  Another aspect of the present invention is a dispensing method in which a liquid containing a biological sample is dispensed to an object to be discharged with a dispensing needle, and the dispensing needle is preliminarily set according to an input desired spot size. It is fixed by moving in a substantially vertical direction with respect to the discharge target surface of the discharge target body so as to be a predetermined distance, and the liquid held by the dispensing needle is pressed by pressing means. The droplet is gradually grown while observing the positional relationship between the tip of the droplet formed on the tip of the dispensing needle and the discharge target surface of the discharge target object with an optical observation means. And determining the distance between the tip of the droplet and the ejection target surface from the positional relationship, and when the distance reaches a predetermined value at which the tip of the droplet can contact the ejection target, A biological sample is obtained by bringing the tip of the droplet into contact with the discharge target body. A liquid containing moving to the discharge object, there is provided a dispensing method.

  According to this, after the tip of the dispensing needle is moved and fixed to a position that coincides with the length in the drop direction of the droplet to be formed, the liquid held by the dispensing needle is pressed by the pressing means. Therefore, it is possible to avoid contamination by dropping the liquid droplets at an unplanned place or timing. In addition, the tip of the dispensing needle is not brought into contact with the object to be ejected (without contact), the tip of the liquid droplet is brought into contact with the object to be ejected, and the liquid droplet is transferred to the object to be ejected by the surface tension of the liquid droplet Therefore, it is possible to avoid variation in spot size due to the dispensing needle coming into contact with the discharge target object. Furthermore, the positional relationship between the tip of the dispensing needle and the ejection target surface is observed by an optical observation means such as a CCD (Charge Coupled Devices), and the distance between the tip of the dispensing needle and the ejection target surface is calculated. 0, that is, the amount of droplets dispensed to the ejection target object can be appropriately and easily controlled by stopping the pressing when the leading end of the droplet reaches a predetermined value capable of contacting the ejection target body. It becomes possible.

  Another aspect of the present invention provides a method for producing a microarray, in which a microarray is produced using the above dispensing method. According to this, since the microarray is manufactured using the above dispensing method, contamination during the microarray manufacturing can be avoided, variation in spot size can be reduced, and the accuracy and yield of the microarray can be improved. Can be made.

  Hereinafter, the dispensing technique of the present invention will be described with reference to the drawings.

  FIG. 1 is an explanatory diagram for schematically explaining the dispensing technique of the present invention.

  As shown in FIG. 1A, the distal end of the dispensing needle 102 can be moved by a z-axis direction moving unit 202 described later in a substantially vertical direction (z-axis direction) with respect to the discharge target surface of the discharge target object. It is configured.

  As shown in FIG. 1B, first, the tip of the dispensing needle 102 is moved by a predetermined distance d and fixed. Here, the distance d is determined in accordance with the size of the spot size to be formed or the amount of liquid to be discharged for forming one spot. Specifically, for example, a table is created by experimentally measuring the relationship between the droplet size (length in the drop direction) and the spot size in advance, and it is desired to form the table by referring to this table. The distance d corresponding to the spot size is determined. Alternatively, instead of the size of the droplets, measure the relationship between the liquid extrusion amount and the spot size in advance, create a table, determine the extrusion amount with reference to the table from the spot size you want to form, and extrude The distance d may be determined by calculating the size (length in the dropping direction) of the droplet 20 from the amount, the diameter of the dispensing needle 102, and the like. More specifically, the droplet 20 expected to be formed from the extrusion amount and the diameter of the dispensing needle 102 is approximated to a sphere, and the length of the droplet 20 in the falling direction, that is, the distance d is calculated. You may decide.

  Next, as shown in FIG. 1C, the liquid is gradually pushed out (exuded) to the tip of the dispensing needle 102 in the dispensing unit by a pressing mechanism 106 (pressing means) described later. Next, as shown in FIG. 1 (d), the droplet 20 formed at the tip of the dispensing needle 102 is grown until the tip of the droplet 20 reaches the discharge target surface of the discharge target object 10. When the liquid contacts the surface to be ejected, liquid extrusion is stopped almost simultaneously. Then, as shown in FIG. 1 (e), the droplet 20 moves to the discharge target object 10 due to the surface tension of the liquid, and a desired spot on the discharge target object 10 as shown in FIG. 1 (f). A spot of size is formed.

  As described above, in the present invention, after the dispensing needle 102 and the discharge target body 10 are positioned, the liquid held in the dispensing unit is pressed to form a droplet formed at the tip of the dispensing needle 102. The liquid is dispensed to the discharge target object 10 by growing 20 and bringing the tip of the droplet 20 into contact. Therefore, in the present invention, the liquid can be dispensed to the ejection target body 10 without bringing the tip of the dispensing needle 102 into contact with the ejection target body 10. Further, when the dispensing needle 102 is moved, no droplet is formed at the tip of the dispensing needle 102, so there is no risk of the droplet falling during the movement. In addition, since the liquid held in the dispensing unit is supplied to the dispensing needle 102, excess liquid does not adhere to the periphery (outer periphery) of the distal end of the dispensing needle 102.

  Here, the biological sample contained in the liquid discharged from the dispensing needle is not particularly limited. For example, in addition to biomolecules such as proteins and nucleic acids, artificially synthesized oligonucleotides, polynucleotides, It also includes analogs such as oligopeptides, polypeptides, PNA (peptide nucleic acids).

  The discharge target body 10 is not particularly limited, and examples thereof include glass, a film, and a synthetic resin (polyethylene, polypropylene, polystyrene, etc.). It is preferable that a lyophilic process is performed on at least a spot formation scheduled position on such a discharge target object 10. Thereby, when the tip of the droplet 20 comes into contact with the discharge target object 10, the transition of the droplet 20 to the discharge target object 10 is promoted.

Next, an embodiment of the present invention will be specifically described.
FIG. 2 is a schematic diagram showing the overall configuration of the dispensing apparatus of the present invention, and FIG. 3 is a block diagram for explaining an example of the dispensing apparatus of the present invention. Hereinafter, the present invention will be described with reference to FIGS. 2 and 3.

  As shown in FIGS. 2 and 3, the dispensing device 1 includes a dispensing unit 100 including a pressing mechanism 106 (pressing unit) that presses a liquid containing a biological sample, the dispensing unit 100, and the discharge target object 10. Is formed at the tip of the dispensing needle 102, and a positioning mechanism configured to be relatively movable in the xyz axis direction, a pressure control unit 302 that controls the amount of liquid discharged from the dispensing unit 100, and the dispensing needle 102. The detection unit 400 is mainly configured to detect the positional relationship between the liquid droplets to be discharged and the discharge target object 10.

  As shown in FIG. 3, the dispensing unit 100 includes a dispensing needle 102, a holding unit 104 that holds a liquid containing a biological sample (also referred to as a sample solution), and a pressing mechanism 106 that presses the liquid. Yes. The dispensing needle 102 and the holding unit 104 are separately configured in the present embodiment, but may be formed integrally. Moreover, it is preferable that the tip of the dispensing needle 102 is subjected to a liquid repellent treatment. Thereby, it becomes possible to promote the transfer of the droplet 20 to the discharge target object 10.

  The pressing mechanism 106 is not particularly limited as long as it can press the sample solution held in the dispensing unit 100, but forms a droplet of a desired size at the tip of the dispensing needle 102. In order to achieve this, it is desirable that the amount of liquid extruded to the tip of the dispensing needle 102 can be accurately adjusted. For example, the pressing mechanism 106 may be configured so that the amount of liquid to be extruded can be adjusted in a minute amount by moving a plunger provided in the dispensing unit 100 by a feed screw mechanism such as a micrometer. In addition, automatic control is possible by configuring the feed screw mechanism to be driven by an actuator.

  The positioning mechanism configured to be able to relatively move the dispensing unit 100 and the discharge target body 10 in the xyz axis direction includes an x axis direction moving unit 204, a y axis direction moving unit 206, and a z axis direction. The moving unit 202 includes an x-axis direction movement control unit 304, a y-axis direction movement control unit 306, and a z-axis direction movement control unit 308 that control driving of each moving unit. The z-axis direction moving unit 202 corresponds to the first moving unit, and the x-axis direction moving unit 204 and the y-axis direction moving unit 206 correspond to the second moving unit.

  The x-axis direction movement unit 204 connects an x-axis direction drive motor (for example, a stepping motor) and the carrier 108 that transports the dispensing unit 100 with a drive belt, and a drive signal supplied from the x-axis direction movement control unit 304. It is configured to drive according to the above. When the x-axis direction moving unit 204 is driven, the carrier 108 moves in the x-axis direction.

  The y-axis direction moving unit 206 can move a moving work table (stage) on which the discharge target object 10 is placed in the y-axis direction by a feed screw system driven by a y-axis direction driving motor (for example, a stepping motor). It is configured.

  The z-axis direction moving unit 202 can move the fixed bar 208 that fixes the carrier 108 that transports the dispensing unit 100 in the z-axis direction by a feed screw system driven by a z-axis direction drive motor (for example, a stepping motor). It is configured.

  The x-axis direction movement control unit 304 and the y-axis direction movement control unit 306 are input by the input unit 600, for example, the xy direction such as the spot formation start position on the discharge target object 10 and the size of the discharge target object 10. In accordance with the relative positioning information, a drive signal is sent to each of the x-axis direction moving unit 204 and the y-axis direction moving unit 206 to move the dispensing needle 102 to a predetermined position on the discharge target body 10 in a substantially horizontal direction.

  The z-axis direction movement control unit 308 refers to the information stored in the storage unit 500 according to the spot size to be formed or the amount of liquid to be ejected input from the input unit 600, determines the movement distance, and determines the predetermined distance. A drive signal is sent to the z-axis direction moving unit 202 so as to move.

  In the storage unit 500, for example, a table created by experimentally measuring the relationship between the spot size and the length in the drop direction of the droplet (corresponding to the distance d) or the relationship between the spot size and the liquid extrusion amount is stored. A table created by experimental measurement is stored.

  The pressing control unit 302 controls the amount of liquid discharged from the dispensing unit 100 (size of the droplet 20). Specifically, for example, after the distal end position of the dispensing needle 102 is moved by a predetermined distance by the z-axis direction moving unit 202 and fixed, the pressing control unit 302 transmits a driving signal to the pressing mechanism 106. Further, a stop signal is transmitted to the pressing mechanism 106 so as to stop the pressing in accordance with the positional relationship information between the droplet 20 and the ejection target 10 detected by the detecting unit 400 described later.

  The detection unit 400 detects the positional relationship between the droplet 20 formed at the tip of the dispensing needle 102 and the discharge target object 10. Specifically, the detection unit 400 includes an observation unit 402 that observes the droplet 20 and the surface of the discharge target object 10, and the tip of the droplet 20 and the discharge target based on the optical image observed by the observation unit 402. An image recognition unit 404 that extracts feature points corresponding to the body ejection target surface, and a distance between the feature points extracted by the image recognition unit 404 is calculated to determine whether or not the distance matches a predetermined value. Part 406.

  For the observation unit 402, for example, an optical observation unit such as a CCD (Charged Coupled Device) camera is used. The optical image captured by the observation unit 402 is converted into a digital signal by an AD converter, and the converted digital signal is temporarily stored in a memory as optical image data.

  The image recognition unit 404 extracts feature points corresponding to the planned position where the tip of the droplet 20 and the tip of the droplet 20 on the ejection target surface come into contact from the accumulated optical image data. The distance between the feature points is calculated from the feature points, and the calculated parameters are transmitted to the determination unit 406.

  The determination unit 406 compares this parameter (distance between the tip of the droplet 20 and the ejection target surface) with a reference value, and if the parameter matches the reference value, a stop signal is sent to the press control unit 302. .

  Here, the reference value may be, for example, the distance 0 at the moment when the tip of the droplet 20 contacts the ejection target surface, and the time lag when the stop signal is sent and the pressing is stopped, and the growth rate of the droplet 20. It is good also as a value calculated in consideration.

  Next, the flow of the dispensing method of this embodiment will be described with reference to FIG. FIG. 4 is a flowchart for explaining the flow of the dispensing method.

  First, xy coordinates of spotting start coordinates are input from the input unit 600 (S1). Based on the input coordinates, the x-axis direction movement control unit 304 and the y-axis direction movement control unit 308 send drive signals to the x-axis direction movement unit 204 and the y-axis direction movement unit 206, respectively. It moves to the spotting start position on the ejection target body 10 (S2).

  Next, a desired spot size is input from the input unit 600 (S3). A table of the spot size and distance d (the distance between the drop direction of the droplet and the distance to the ejection target surface) stored in the storage unit 500 is referred to determine the distance d corresponding to the input spot size (S4). , S5).

  The z-axis direction movement control unit 306 transmits a drive signal to the z-axis direction movement unit 202, and the dispensing unit 100 is located at a position where the distance between the tip of the dispensing needle 102 and the droplet discharge target surface is a distance d. Is moved (S6). Here, the movement of the dispensing unit 100 in the xy direction and the movement in the z-axis direction may be reversed.

  When the position of the dispensing unit 100 is fixed, the pressing control unit 302 instructs the pressing mechanism 106 to slowly extrude the liquid (S7). The distance relationship between the droplet 20 and the discharge target object 10 is imaged by an observation means (observation unit 402) such as a CCD camera, and the image recognition unit 404 extracts feature points from the obtained optical image, and the feature points. A distance L between the tip of the droplet 20 and the ejection target surface is calculated from the distance. Next, in the determination unit 406, it is determined whether or not the distance L matches the predetermined reference value as described above. If not, the growth of the droplet is observed. An instruction is sent to the pressing control unit 302 to stop driving the pressing mechanism 106 (S10). When the droplet 20 comes into contact with the target object 10 to be discharged, it moves to the target object 10 side due to surface tension, and a spot of a desired size is formed.

  Note that the timing at which the pressing control unit 302 transmits the driving signal to the pressing mechanism 106 occurs until the driving signal from the z-axis direction movement control unit 306 is transmitted and the z-axis direction moving unit 202 actually completes the movement. In consideration of the time lag, the pressing control unit 302 may transmit the driving signal to the pressing mechanism 106 after a predetermined time after the driving signal from the z-axis direction movement control unit 306 is stopped.

  According to the present embodiment, after the dispensing unit 100 reaches a predetermined position on the discharge target object 10, droplets are formed by the pressing mechanism 106 provided in the dispensing unit 100. It is possible to avoid contamination of other spots by dropping droplets at unscheduled locations and timings when moving 100 or the like. In addition, since the dispensing needle 102 is not brought into contact, and the droplet formed at the tip of the dispensing needle 102 is brought into contact with the ejection target surface of the ejection target object 10 and dispensed, the dispensing needle 102 is ejected. Inconveniences such as spot size variation and biological sample denaturation due to contact with the body 10 can be avoided. In this embodiment, the sample solution is held in the dispensing unit 100 in advance, and it is not necessary to immerse the dispensing needle 102 in the sample solution in order to replenish the dispensing needle 102 with the sample solution. Variation in spot size due to excess liquid adhering to the outer periphery of the tip of the needle 102 can be avoided.

  In addition, the size of the droplet formed on the dispensing needle 102 may vary depending on the surrounding environment such as the property (for example, viscosity) and humidity of the sample solution to be used. Since the droplet 20 is grown while confirming the size of the droplet 20, the droplet 20 can be reliably brought into contact with the discharge target object 10. Therefore, it is possible to avoid problems such as the case where the droplet 20 cannot contact the discharge target object 10 and is not dispensed.

  In the above embodiment, the case where one dispensing unit 100 is used has been described as an example, but a plurality of dispensing units may be used simultaneously.

  Moreover, in the said embodiment, although the dispensing unit 100 moved the z-axis direction moving part 202 by the drive signal from the z-axis direction movement control part 306, and moved to the z-axis direction, it is not limited to this. For example, as shown in FIG. 5, the positioning member 210 is fixed in advance to a position where the dispensing unit 100 is to be manually moved, and the fixing bar 208 is moved to the position where the positioning member 210 is fixed. Positioning in the z-axis direction may be performed.

  In the above-described embodiment, the distance between the tip of the droplet 20 and the discharge target object 10 is detected by the detection unit 400 using an observation unit such as a CCD. However, the present invention is not limited to this. You may observe the distance with the target object 10 by visual observation, and may stop a press manually. Alternatively, the stop timing of pressing may be measured by using a laser light source and a light receiving sensor and a change in received light intensity when a droplet crosses the laser light source generated from the laser light source.

  In the above embodiment, the formation of droplets is observed every time, but the amount of liquid used at the time of the first droplet formation is measured, and the amount of liquid used at the first time is discharged after the second time. In this manner, the pressing control unit 302 may be instructed. According to this, it becomes possible to shorten the time required for the second and subsequent dispensing, and work efficiency can be improved.

  The dispensing apparatus of the present invention can be used as a microarray manufacturing apparatus, and a microarray can be manufactured using the dispensing method of the present invention.

FIG. 1 is an explanatory diagram for schematically explaining the dispensing technique of the present invention. FIG. 2 is a schematic diagram showing the overall configuration of the dispensing apparatus of the present invention. FIG. 3 is a block diagram for explaining an example of the dispensing device of the present invention. FIG. 4 is a flowchart for explaining the flow of the dispensing method. FIG. 5 is a diagram for explaining an example of a method for moving the dispensing unit 100 in the z-axis direction.

Explanation of symbols

10 target object, 20 droplets, 100 dispensing unit, 102 dispensing needle, 104 holding unit, 106 pressing mechanism, 108 carrier, 202 z-axis direction moving unit, 204 x-axis direction moving unit, 206 y-axis direction moving , 208 fixed bar, 210 positioning member, 302 pressure control unit, 304 x-axis direction movement control unit, 306 z-axis direction movement control unit, 308 y-axis direction movement control unit, 400 detection unit, 402 observation unit, 404 image recognition Part, 406 discrimination part, 500 storage part, 600 input part

Claims (13)

A holding unit for holding a liquid containing a biological sample, a pressing means for pressing the held liquid, and a dispensing needle capable of discharging a droplet,
A dispensing unit capable of forming a droplet at the tip of the dispensing needle by pressing the liquid by the pressing means;
The distance between the distal end of the dispensing needle and the ejection target surface of the ejection target body in a direction substantially perpendicular to the ejection target surface is the drop direction of the liquid droplet that is to be formed at the distal end of the dispensing needle. First moving means for moving and fixing the tip position of the dispensing needle in the substantially vertical direction so as to be substantially the same as the length of
A dispensing device comprising:   2. The apparatus according to claim 1, further comprising a second moving unit that moves the dispensing unit to a spot formation position on the discharge target object relatively horizontally with a discharge target surface of the discharge target object. Dispensing device.   The dispensing device according to claim 1 or 2, wherein a liquid repellent treatment is applied to a tip of the dispensing needle.   The dispensing apparatus according to any one of claims 1 to 3, wherein a lyophilic process is performed on the discharge target object.   The dispensing apparatus according to any one of claims 1 to 4, wherein the pressing means is means for pressing a liquid by moving a plunger provided in the dispensing unit by a feed screw mechanism.   The dispensing apparatus according to claim 1, wherein the feed screw mechanism is driven by an actuator. An input means for inputting a desired spot size;
A storage means in which the relationship between the spot size and the distance between the tip of the dispensing needle and the discharge target object is stored in a readable manner;
The first moving means for transmitting a first drive signal to the first moving means so as to move the distance read from the storage means in response to the input of the spot size to the input means. Movement control means for controlling;
A pressure control means for transmitting a second drive signal to the pressure means after the tip position of the dispensing needle has been moved and fixed to the distance;
Detecting means for detecting a positional relationship between a droplet formed at the tip of the dispensing needle and a discharge target surface of the discharge target body;
The pressing control means further transmits a stop signal to stop the driving according to the positional relationship between the droplets detected by the detecting means and the ejection target surface;
The dispensing apparatus according to any one of claims 1 to 6.   The detection means comprises: an observation means for observing a droplet formed at the tip of the dispensing needle and a discharge target surface of the discharge target; a droplet tip based on a light image observed by the observation means; The dispensing apparatus according to claim 7, further comprising: a determination unit that calculates a distance to the ejection target surface, compares the distance with a predetermined value, and determines whether or not the distance matches the predetermined value.   The dispensing device according to claim 7 or 8, wherein the pressing control means transmits the second drive signal after a predetermined time after the first drive signal is stopped.   A microarray manufacturing apparatus comprising the dispensing apparatus according to claim 1. A dispensing method for dispensing a liquid containing a biological sample to an object to be discharged with a dispensing needle,
The distance between the distal end of the dispensing needle and the ejection target surface of the ejection target body in a direction substantially perpendicular to the ejection target surface is the drop direction of the liquid droplet that is to be formed at the distal end of the dispensing needle. The tip position of the dispensing needle is moved and fixed so as to match the length of
The liquid tip held by the dispensing needle is exuded to the tip of the dispensing needle by being pressed by a pressing means, and the droplet formed at the tip of the dispensing needle is gradually grown, whereby the tip of the droplet A dispensing method in which pressing is stopped substantially simultaneously with contact with the discharge target body, and a liquid containing a biological sample is moved to the discharge target body. A dispensing method for dispensing a liquid containing a biological sample to an object to be discharged with a dispensing needle,
The dispensing needle is moved and fixed in a substantially vertical direction with respect to the discharge target surface of the discharge target object so as to be a predetermined distance according to the input desired spot size,
The liquid held by the dispensing needle is exuded to the tip of the dispensing needle by pressing with a pressing means, and the tip of the droplet formed at the tip of the dispensing needle and the discharge target surface of the discharge target object While observing the positional relationship with the optical observation means, gradually grow the droplets,
Find the distance between the droplet tip and the discharge target surface from the positional relationship,
When the distance reaches a predetermined value at which the tip of the droplet can contact the discharge target body, the pressing is stopped,
A dispensing method in which a liquid containing a biological sample is moved to the discharge target body by bringing the tip of the droplet into contact with the discharge target body.   The manufacturing method of a microarray which manufactures a microarray using the dispensing method of Claim 11 or Claim 12.

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