JP4033468B2 - Nozzle tip position measuring device and spotting device using the same - Google Patents

Description

  The present invention relates to a nozzle tip position measuring device that measures the position of a nozzle tip of a spotting device that drops a minute amount of liquid onto a substrate such as a glass plate.

  In recent years, in the medical field, treatments and drugs using genes (DNA) have been developed. For example, what is called a “DNA chip” in which thousands to tens of thousands of copies of different genes are sampled on a substrate made of a glass plate, a synthetic resin plate, or the like, or thousands on a substrate such as a semiconductor A so-called “protein synthesis chip” that has a reaction vessel (microwell) of from tens to tens of thousands of cells, and performs a reaction by putting a sample (synthesizing protein) or an enzyme into the microwell. What is called has been proposed (see, for example, cited document 1).

  In the spotting apparatus used for manufacturing the chip, in order to drop a small amount of reagent on the substrate, a type in which a pin with a sharp tip such as a Stanford type is brought into contact with the pio chip and spotted is the mainstream. However, non-contact nozzles that do not come into contact with biochips have been developed due to problems such as damage to biochips and pins, and the occurrence of contamination (contamination) derived therefrom. In handling these contact-type pins and non-contact nozzles, it is important to accurately obtain the positional relationship between the tip, the pin, and the nozzle in order to perform an accurate spot.

  In such a spotting apparatus, the present applicant has already proposed a spotting apparatus in which a spotting head is provided in a moving means that can move relative to each other in the XYZ three-axis directions orthogonal to each other, and the head is controlled based on position information from the imaging means ( For example, see Patent Document 2).

There are the following methods for measuring the tip position of a pin or nozzle in a conventional spotting apparatus.
A method for correcting the deviation of the actual spot position with respect to the target position by actually making a test strike on the X and Y axes (horizontal direction).
・ For the Z axis (vertical direction), the nozzle is visually lowered to the vicinity of the tip, and the zero point is adjusted by making the nozzle contact the tip by fine adjustment.
Others • A method that uses a micromanipulator to roughly align the position visually, and then finely adjust the position manually so that it is within the microscope field of view.
In a semiconductor manufacturing apparatus, a method of acquiring tool tip position information by installing an infrared camera or the like for photographing a tool tip separately from a camera for photographing a substrate.

JP 2000-236876 A JP2003-149094

However, the conventional nozzle tip position measuring method has the following problems.
-Accurate position information on the micrometer order cannot be obtained.
・ If the nozzle tip is physically contacted, there is a risk of damaging the nozzle tip, which may cause contamination.
・ When performing manually, there is a high possibility that it will take time and position data will vary.
When a separate camera system is used, a camera for measuring the nozzle tip position is used only for that purpose, which is problematic in terms of cost.
When measuring the position of the nozzle tip with a laser measuring instrument or the like, depending on the measuring instrument to be used, if the laser spot diameter is larger than the nozzle tip, measurement cannot be performed or measurement accuracy is deteriorated.

  The present invention has been made in view of the above problems, and provides a nozzle tip position measuring device capable of accurately measuring accurate position information on the order of micrometers without causing damage to the nozzle tip or contamination. An object of the present invention is to obtain a nozzle tip position measuring device that is free from cost problems.

The nozzle tip position measuring device according to the invention described in claim 1 includes a moving means capable of relative movement in the XYZ three-axis directions orthogonal to the XY plane of the base ,
A nozzle provided in the moving means;
A nozzle tip position measuring device for measuring a position of a nozzle tip in a spotting device provided with an imaging means for photographing the base,
An optical system for measuring the position of the nozzle tip installed on the base, and a position measuring means for measuring a positional deviation from a reference position of the nozzle tip,
The optical system includes a reference area hole formed in a reference plate, an optical path that guides an image of the reference area hole from the bottom surface direction of the reference plate to the imaging means, and the imaging means disposed in the optical path. A lens for focusing on the bottom surface position of the reference plate in which the reference area hole is formed at a fixed focal length of
The position measuring means inserts the nozzle tip into the reference area hole, moves the nozzle tip in the Z-axis direction on the captured image including the reference area hole, and adjusts the focus by moving the nozzle tip in the Z-axis direction. The reference position is acquired, and the positional deviation in the X-axis and Y-axis directions of the nozzle tip with respect to the center of the reference area hole at the reference position in the Z-axis direction is measured .

The spotting apparatus according to the invention described in claim 2 is a reagent arrangement in each microwell of a microwell assembly in which a plurality of microwells are integrated on a substrate, or a reagent from within each microwell. A spotting device for collecting
Moving means capable of relative movement in the XYZ 3-axis directions orthogonal to each other with respect to the base of the XY plane on which the microwell assembly is placed;
A spotting nozzle provided in the moving means, for arranging a reagent in the individual microwell or collecting a reagent from the individual microwell;
Imaging means for photographing the arrangement of the plurality of microwells from above;
Arithmetic means for calculating position information of individual microwell images from image information obtained by the imaging means;
A spotting apparatus comprising: control means for controlling the moving means and spotting nozzle means based on the position information calculated by the computing means;
An optical system for measuring a nozzle tip position installed on the base; and a position measuring means for measuring a positional deviation from a reference position of the nozzle tip;
The optical system includes a reference area hole formed in a reference plate, an optical path that guides an image of the reference area hole from the bottom surface direction of the reference plate to the imaging means, and the imaging means disposed in the optical path. A lens for focusing on the bottom surface position of the reference plate in which the reference area hole is formed at a fixed focal length of
The position measuring means inserts the nozzle tip into the reference area hole, moves the nozzle tip in the Z-axis direction on the captured image including the reference area hole, and adjusts the focus by moving the nozzle tip in the Z-axis direction. characterized in that acquires a reference position, is intended to be used as calibration information for the Z-axis direction of the X axis of the nozzle tip relative to the center of the reference area hole in the reference position, Y-axis direction positional deviation measurement to the control unit It is what.

  As described above, the present invention has an effect that it is possible to obtain a nozzle tip position measuring device capable of obtaining accurate position information in the order of micrometers without causing damage to the nozzle tip or contamination.

In the present invention, a moving means capable of relative movement in XYZ triaxial directions perpendicular to each other with respect to the XY plane of the base , a nozzle provided in the moving means, and an imaging means for photographing the base are provided. Nozzle tip position measuring device that measures the position of the nozzle tip in the spotting device, and measuring the positional deviation of the nozzle tip from the reference position and an optical system for measuring the nozzle tip position installed on the base The optical system includes a reference area hole formed in a reference plate, an optical path for guiding an image of the reference area hole from the bottom surface of the reference plate to the imaging means, and the optical path. A lens for focusing on the position of the bottom surface of the reference plate in which the reference area hole is formed at a fixed focal length of the imaging means disposed therein; Provided by the captured image at the time of moving the nozzle tip to a predetermined reference position in the reference area bore, for measuring the three directions of the deviation from a reference position of the nozzle tip, Ya breakage of the nozzle tip Accurate position information in the micrometer order can be accurately measured without occurrence of contamination.

The apparatus of the present invention can be incorporated in addition to, for example, an existing spotting apparatus. That is, a spotting apparatus for arranging a reagent in each microwell of a microwell assembly in which a plurality of microwells are integrated on a substrate or collecting a reagent from each microwell, A moving means capable of relatively moving in the XYZ three-axis directions perpendicular to each other with respect to the base of the XY plane on which the well integrated body is placed, and provided in the moving means, and a reagent is arranged in each individual microwell A spotting nozzle that collects a reagent from within the microwell, an imaging unit that images the array of the plurality of microwells from above, and an arithmetic unit that calculates position information of each microwell image from image information obtained by the imaging unit And controlling the moving means and the spotting nozzle means based on the position information calculated by the calculating means. In spotting device provided with a control means, further comprising a position measuring unit for measuring an optical system for the nozzle tip position measurement which is mounted on the base, a positional deviation from a reference position of the nozzle tip, the The optical system includes a reference area hole formed in the reference plate, an optical path for guiding an image of the reference area hole from the bottom surface direction of the reference plate to the image pickup means, and the image pickup means disposed in the optical path. And a lens for focusing on the bottom surface position of the reference plate in which the reference area hole is formed at a fixed focal length, and the position measuring means inserts the nozzle tip into the reference area hole, A reference position in the Z-axis direction of the nozzle tip is obtained by moving the nozzle tip in the Z-axis direction on the captured image including X-axis of the nozzle tip relative to the center of the reference area hole in the reference position in the Z-axis direction, measures the positional deviation in the Y-axis direction, using the positional displacement information output from the measuring means as the calibration information of the control means. Thereby, it is possible to obtain a spotting device capable of accurately measuring accurate position information on the order of micrometers without causing breakage of the nozzle tip or mixing of foreign matters.

As an imaging means for guiding an image from the bottom surface direction of the reference plate of the reference area hole by the optical path of the present invention, a new imaging means may be provided, or an existing imaging means may be used . Thereby , a nozzle tip position measuring device having no problem in cost is obtained.

That is, as the optical path, the input side focus is used as a reference area hole , and the position of an optical system such as a mirror or a lens is adjusted so as to match the bottom surface of the reference plate in which the reference hole is formed. The image of the reference area hole may be guided to the imaging means arranged side by side. It does not matter if a mirror is inserted in the middle lens part and the optical path is bent in the horizontal direction.

When measuring the position of the nozzle tip in the apparatus of the present invention, the reference area hole is positioned according to the arrangement of the optical device, the nozzle is lowered, and the nozzle tip is inserted into the reference area hole. The vertical position of the nozzle tip can be obtained by matching the reference plate and the focus position of the nozzle tip on the camera image of the reference area hole . By processing the area camera image, it is obtained by the relative position of the nozzle tip with respect to the reference hole, and by calculating the absolute position of the reference hole and the relative position of the nozzle tip, position information in the horizontal plane of the nozzle tip is obtained. be able to.

  The apparatus of the present invention using this existing image pickup means can be additionally incorporated into the existing apparatus, and an apparatus having no problem in cost can be obtained.

  In the present invention, the position of the nozzle tip in the three directions (X axis, Y axis, and Z axis) is set relative to the reference plate by an existing image processing apparatus without contact with a sensor or replacement of a lens, a camera, or the like. It can be measured as a position. For example, in the Z-axis direction, the measurement accuracy may deteriorate due to the performance of the camera and lens, but the existing image processing camera can be diverted to the measurement of the nozzle tip position. Since it can be performed, there is an advantage that the nozzle tip is not damaged. As for the XY axis, it is not necessary to pursue the accuracy of the nozzle or camera mounting position itself as long as the accuracy of the reference hole is obtained, so there is no need to add expensive and highly accurate parts and equipment, and the cost performance is excellent. There are advantages.

  In addition, since it is not necessary to remove existing equipment every time measurement is performed, measurement with high reproducibility can be easily performed.

As described above, the following effects can be achieved by this apparatus. By using the reference area hole , the relative position of the nozzle tip with respect to the reference area hole is measured, so that high measurement accuracy can be obtained even if there is a slight displacement of the optical system. In addition, since the position of the nozzle tip can be accurately measured, the position of the nozzle tip can be adjusted without performing trial strikes. Thereby, a stable spot can always be performed. Furthermore, since the optical system is incorporated outside the objective lens, it is possible to save the trouble of replacing the lens each time the nozzle tip is measured, and to ensure high reproducibility. In addition, since an existing image processing camera can be used, there is no need to add a device and the cost performance is excellent. Furthermore, since the position of the nozzle tip is measured in a non-contact manner, there is a low possibility of foreign matter contamination due to nozzle breakage or the like.

  FIG. 1 is an explanatory view showing a configuration of a spotting apparatus incorporating a measuring apparatus according to the present invention. As shown in the figure, a pair of rails 12 and 13 rises so as to surround two opposite parallel sides of the base 11 and can be moved in a state of being horizontally mounted on the rails 12 and 13 as the y arm portion. A stage 14 is provided. The y stage 14 is equipped with a linear motor (not shown) for moving on a pair of rails, and can obtain high positional accuracy.

  The y stage 14 is provided with an x stage 16 that is movable along an x arm portion 15 that extends in a direction perpendicular to the rails 12 and 13. The x stage 16 is also equipped with a linear motor (not shown) for moving on the x arm portion 15, and high positional accuracy can be obtained. The x stage 16 is provided with a z stage 18 that is movable along a z arm portion 17 that extends in the height direction. The z stage 18 is moved on the z arm 17 by a servo motor and a ball screw (not shown).

  In the z stage 18, a spotting nozzle 21 for arranging a reagent in a microwell formed in a microwell assembly 19 placed and fixed on the base 11 and collecting the reagent from each microwell, and this An area camera 22 arranged in parallel with the nozzle 21 is provided.

  On the base 11 on which the microwell assembly 19 is placed and fixed, a cleaning unit 2 for cleaning the nozzle, a drying unit 3 for drying the cleaned nozzle, and a reagent container 5 are arranged in parallel. In addition, an optical system 20 for measuring the nozzle tip position is provided side by side.

In the optical system 20 for measuring the nozzle tip position, a reference hole 23 as a reference area hole into which the spotting nozzle 21 is inserted, and the spotting nozzle 21 is formed immediately below the area camera 22 when the spotting nozzle 21 is inserted into the reference hole 23. The focus hole 24 is formed.

  FIG. 2 is an explanatory diagram showing the configuration of an embodiment of an optical system for measuring the nozzle tip position. As shown in FIG. 2, mirrors 25 and 26 are provided directly below the reference hole 23 and the focus hole 24, and lenses 27 and 28 are mounted so as to connect the optical paths of the two mirrors. The focus on the nozzle side of the optical system is adjusted to the bottom surface of the reference plate. Usually, the focal length of the area camera 22 is fixed for photographing a chip (glass plate), but in order to maintain the focal length and use it for measuring the nozzle tip without changing the focus, the optical path distance is used. And lenses 27 and 28 are adjusted in advance.

Thereby, when the nozzle is replaced or at any other timing, the position of the nozzle tip in the three-axis direction can be measured by the following procedure. The specific operation is as follows.
(1) Position the reference position of the linear stage so that the position of the nozzle is aligned with the position of the reference hole of the optical system. Alternatively, the nozzle tip is inserted into the reference hole by direct visual inspection.

  (2) In the area camera image, lower the Z axis so that the tip of the nozzle is in focus with respect to the reference plate. FIG. 3 is an explanatory diagram showing a state of focusing for determining the position of the nozzle tip in the Z-axis direction. FIG. 3A shows a state before the nozzle tip is inserted into the reference hole and an image at that time. Indicates the state in which the nozzle tip is inserted at the reference position of the reference hole and the image at that time. As shown in the figure, the movement in the Z-axis direction is performed, and the nozzle tip and the bottom surface of the reference plate substantially coincide with each other at the focused position as shown in FIG. Can be acquired.

(3) With the nozzle tip in focus, the displacement of the nozzle tip in the X-axis and Y-axis directions with respect to the reference hole is measured by image processing. FIG. 4 is an explanatory diagram showing the deviation of the nozzle tip in the X-axis and Y-axis directions. As shown in the figure, the Z axis is determined as the reference position of the reference hole by adjusting the optical path distance and the lenses 27 and 28 in advance without changing the focus while maintaining the focal length of the area camera. The point where the focus is obtained is the Z-axis reference position. Further, the deviation in the X-axis and Y-axis directions from the reference position of the nozzle tip (for example, the center of the reference area hole ) when focus is achieved can be controlled as calibration information.

It is explanatory drawing which shows the structure of the spotting apparatus incorporating the measuring device by this invention. It is explanatory drawing which shows the structure of one Example of the optical system for nozzle tip position measurement. It is explanatory drawing which shows the state of the focus adjustment for the position determination of the Z-axis direction of the nozzle tip. It is explanatory drawing which shows the shift | offset | difference of the X-axis direction of a nozzle front-end | tip, and a Y-axis direction.

Explanation of symbols

2 ... Cleaning unit,
3 ... Drying unit,
5 ... Reagent storage,
11 ... Base,
12 ... Rail,
13 ... Rail,
14 ... y stage
15 ... x arm part,
16 ... x stage,
17 ... z arm part,
18 ... z stage,
19 ... Microwell assembly,
20: Optical system for measuring the nozzle tip position,
21 ... Spotting nozzle,
22 Area camera,
23: Reference hole,
24: Focus hole,
25 ... Mirror,
26 ... Mirror,
27 ... Lens,
28 ... Lens,

Claims (2)

Moving means capable of relative movement in the XYZ triaxial directions perpendicular to the XY plane of the base ;
A nozzle provided in the moving means;
A nozzle tip position measuring device for measuring a position of a nozzle tip in a spotting device provided with an imaging means for photographing the base,
An optical system for measuring the position of the nozzle tip installed on the base, and a position measuring means for measuring a positional deviation from a reference position of the nozzle tip,
The optical system includes a reference area hole formed in a reference plate, an optical path that guides an image of the reference area hole from the bottom surface direction of the reference plate to the imaging means, and the imaging means disposed in the optical path. A lens for focusing on the bottom surface position of the reference plate in which the reference area hole is formed at a fixed focal length of
The position measuring means inserts the nozzle tip into the reference area hole, moves the nozzle tip in the Z-axis direction on the captured image including the reference area hole, and adjusts the focus by moving the nozzle tip in the Z-axis direction. A nozzle tip position measuring apparatus that acquires a reference position and measures a positional deviation of the nozzle tip in the X-axis and Y-axis directions with respect to the center of the reference area hole at the reference position in the Z-axis direction . A spotting device for arranging a reagent in each microwell of a microwell assembly in which a plurality of microwells are integrated on a substrate or collecting a reagent from each microwell,
Moving means capable of relative movement in the XYZ 3-axis directions orthogonal to each other with respect to the base of the XY plane on which the microwell assembly is placed;
A spotting nozzle provided in the moving means, for arranging a reagent in the individual microwell or collecting a reagent from the individual microwell;
Imaging means for photographing the arrangement of the plurality of microwells from above;
Arithmetic means for calculating position information of individual microwell images from image information obtained by the imaging means;
A spotting apparatus comprising: control means for controlling the moving means and spotting nozzle means based on the position information calculated by the computing means;
An optical system for measuring a nozzle tip position installed on the base; and a position measuring means for measuring a positional deviation from a reference position of the nozzle tip;
The optical system includes a reference area hole formed in a reference plate, an optical path that guides an image of the reference area hole from the bottom surface direction of the reference plate to the imaging means, and the imaging means disposed in the optical path. A lens for focusing on the bottom surface position of the reference plate in which the reference area hole is formed at a fixed focal length of
The position measuring means inserts the nozzle tip into the reference area hole, moves the nozzle tip in the Z-axis direction on the captured image including the reference area hole, and adjusts the focus by moving the nozzle tip in the Z-axis direction. characterized in that acquires a reference position, is intended to be used as calibration information for the Z-axis direction of the X axis of the nozzle tip relative to the center of the reference area hole in the reference position, Y-axis direction positional deviation measurement to the control unit A spotting device.

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