JP2018146479A - Spot deposition amount measurement system, spot deposition amount measurement method and program - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a spot deposition amount measurement system or the like capable of accurately grasping the amount of liquid spotted on a material to be spotted.SOLUTION: A spot deposition amount measurement system includes: an imaging device for imaging an object and outputting image data; and an information processing device for generating an unspotted image being an image which is imaged before being spotted on a material to be spotted by a dispensation pipette to capture at least a tip of a chip provided in the dispensation pipette and liquid suspending from the tip, and a spotted image being an image picked up after being spotted on the material to be spotted by the dispensation pipette to capture at least the tip of the chip on the basis of image data outputted from the imaging device, and calculating a spot deposition amount being the capacity of the liquid spotted on the material to be spotted on the basis of the unspotted image and the spotted image.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a spot amount measuring system, a spot amount measuring method, and a program for measuring the volume of a liquid spotted by a dispensing pipette.

Conventionally, in the medical field, biochemical field, and the like, examinations and experiments in which a small amount of liquid such as a reagent or a sample is reacted in a microplate or the like have been performed. The liquid used for the reaction is usually
A spotting tip for sucking and discharging liquid (also simply referred to as a tip) is spotted on the spotted material using a dispensing pipette attached to the tip.

  As a technique related to spotting, Patent Document 1 discloses a detection signal of temperature and humidity related to the vapor pressure of the atmosphere, and evaporation of the liquid in the spotting chip after the liquid is sucked into the spotting chip. Disclosed is a liquid spotting method that corrects the discharge pressure in the initial spotting to be low so that the spotting amount at each spotting is equal in continuous spotting based on the characteristics of the change in vapor pressure related to the accompanying detection signal. Has been.

Japanese Patent No. 3390297

  The present inventors have observed the state of the tip of the tip at the time of spotting. Even when continuous spotting is performed by inhaling more liquid than the amount of spotting once into the tip, It has been found that liquid may adhere to the tip of a later chip. In such a case, even if the amount of liquid discharged from the chip is accurately controlled, the prescribed amount of liquid is not in the spotted material. Therefore, in order to ensure the accuracy of inspections and experiments, it is necessary not only to control the amount of liquid discharged from the chip, but also to accurately grasp the amount of liquid actually spotted on the spotted material. It is.

  The present invention has been made in view of the above, and provides a spotting amount measuring system, a spotting amount measuring method, and a program capable of accurately grasping the amount of liquid spotted on a spotted material. For the purpose.

  In order to solve the above-described problem, a spotted amount measurement system according to one aspect of the present invention is based on an imaging device that images a subject and outputs image data, and dispensing based on the image data output from the imaging device. An image before spotting that is imaged before spotting a spotted material with a pipette and at least a tip portion of a tip provided in the pipetting pipette and a liquid that hangs down from the tip portion; and An image after being spotted on the spotted material by a pipetting pipette, and an image after spotting that is an image in which at least the tip of the tip is shown, is generated, and the image before spotting and the image after spotting are generated. And an information processing device that calculates a spotting amount that is a volume of the liquid spotted on the spotted material.

  In the spotting amount measurement system, the information processing device generates the pre-spotting image and the post-spotting image, and the liquid is not drooping based on the image data output from the imaging device. A reference image, which is an image of the tip of the chip, is generated, a first difference image between the image before spotting and the reference image is generated, and the liquid is reflected from the first difference image. An image processing unit that extracts one region, generates a second difference image between the spotted image and the reference image, and extracts the second region in which the liquid is reflected from the second difference image And a spotting amount calculation unit that calculates the spotting amount based on the extracted first and second regions.

  In the spotting amount measurement system, the image processing unit further includes an image of the chip between the pre-spotting image and the reference image and between the post-spotting image and the reference image. Alignment may be performed based on this.

  In the spotting amount measurement system, the information processing device generates the pre-spotting image and the post-spotting image, and generates a difference image between the pre-spotting image and the post-spotting image, You may have an image processing part which extracts the area | region where the said liquid was reflected from the difference image, and a spotting amount calculation part which calculates the said spotting amount based on the said extracted area | region.

  In the top spotting amount measurement system, the image processing unit may further perform alignment between the pre-dotting image and the post-spotting image based on the chip image.

  In the spotting amount measurement system, the chip includes a chip body that discharges a liquid, and a marker provided on an outer peripheral surface of the chip body, and the imaging device at least places the marker in the field of view. The image processing unit may perform the alignment based on the marker image.

  In the spotting amount measurement system, the image processing unit may perform inclination correction of the pre-dotting image and the post-spotting image based on the marker image.

In the spotting amount measurement system, the imaging device may include a telecentric lens.
The spotting amount measurement system may further include a light source that illuminates at least the field of view of the imaging device.

  A spotting amount measuring method according to another aspect of the present invention is provided at least in the pipetting pipette based on image data acquired by performing imaging before spotting a spotted material with a pipetting pipette. A step of generating an image before spotting which is an image in which the tip of the tip and the liquid dripping from the tip are reflected, and imaging after spotting the spotted material with the pipetting pipette Based on the acquired image data, at least a step of generating an image after spotting that is an image in which the tip portion of the chip is reflected, and the spotted material based on the image before spotting and the image after spotting Calculating a spotting amount which is a volume of the spotted liquid.

  A program according to still another aspect of the present invention is a spotting comprising: an imaging device that images a subject and outputs image data; and an information processing device that performs information processing based on the image data output from the imaging device. In a quantity measurement system, a program to be executed by the information processing apparatus, wherein the imaging apparatus executes imaging before spotting a spotted material with a dispensing pipette, and image data output from the imaging apparatus is generated. A step of generating an image before spotting which is an image in which at least a tip portion of a tip provided in the pipetting pipette and a liquid hanging from the tip portion are reflected; and the spotted material by the pipetting pipette The image is an image in which at least the tip of the chip is captured based on image data output from the imaging device by causing the imaging device to perform imaging after spotting A step of generating a post-image, and a step of calculating a spotting amount, which is a volume of liquid spotted on the spotted material, based on the pre-spotting image and the post-spotting image. It is.

  According to the present invention, an image before spotting in which the tip portion of the chip and the liquid dripping from the tip portion are captured, and an image after spotting in which the tip portion of the chip after spotting is captured, are obtained. It is possible to measure the amount of change in the liquid before and after the spotting, that is, the amount of the liquid substantially spotted on the spotted material, by performing an operation using the image before spotting and the image after spotting. it can. Therefore, it is possible to accurately grasp the amount of liquid spotted on the spotted material.

It is a figure which shows schematic structure of the spotting amount measurement system which concerns on embodiment of this invention. It is a schematic diagram which expands and shows the front-end | tip part of the chip | tip shown in FIG. It is a block diagram which shows schematic structure of the information processing apparatus shown in FIG. It is a flowchart which shows operation | movement of the spotting amount measurement system shown in FIG. It is a schematic diagram for demonstrating operation | movement of a spot amount measurement system. It is a schematic diagram for demonstrating operation | movement of a spot amount measurement system. It is a schematic diagram for demonstrating operation | movement of a spot amount measurement system. It is a schematic diagram for demonstrating operation | movement of a spot amount measurement system. It is a schematic diagram for demonstrating operation | movement of a spot amount measurement system. It is a schematic diagram for demonstrating the modification of the spotting amount measuring method which concerns on embodiment of this invention.

  Hereinafter, a spotting amount measurement system, a spotting amount measurement method, and a program according to an embodiment of the present invention will be described with reference to the drawings. Note that the present invention is not limited to these embodiments. Moreover, in description of each drawing, the same code | symbol is attached | subjected and shown to the same part.

  The drawings referred to in the following description only schematically show the shape, size, and positional relationship so that the content of the present invention can be understood. That is, the present invention is not limited only to the shape, size, and positional relationship illustrated in each drawing. Also, there are cases in which parts having different dimensional relationships and ratios are included between the drawings.

  FIG. 1 is a diagram showing a schematic configuration of a spotted amount measuring system according to an embodiment of the present invention. A spotting amount measuring system 1 shown in FIG. 1 is a system that measures the amount of liquid spotted (spotted quantity) in a spotting operation in which a liquid is spotted on a microplate 2 using a dispensing pipette 3. is there. As shown in the figure, the spotted amount measuring system 1 includes a camera 4 that images a subject and outputs image data, an illumination device 5 that illuminates the field of view of the camera 4, and image data output from the camera 4. And an information processing device 6 for generating an image and calculating a spotting amount based on the image. Note that the lighting device 5 may be omitted when the installation location of the spotted amount measurement system 1 is bright enough to clearly obtain various images described later.

  The microplate 2 is an instrument in which a plurality of wells 21 for arranging reagents and specimens are arranged in a matrix. In this embodiment, the case where the microplate 2 is used as the spotted material will be described. However, the type of the spotted material is not limited to this, and the present invention is applicable to all general-purpose spotted materials. This can be applied when spotting.

  The pipetting pipette 3 has a pipette body 31 and a tip 32 attached to the tip of the pipette body 31. The pipette body 31 includes a syringe and a piston provided in the syringe, and liquid can be sucked or discharged from the tip of the tip 32 by moving the piston along the axial direction of the syringe.

  FIG. 2 is a schematic diagram showing the tip portion of the chip 32 in an enlarged manner. As shown in FIG. 2, in the chip 32, it is preferable to provide a marker 34 for alignment in image processing to be described later with respect to the chip main body 33 that forms a liquid flow path. In the present embodiment, a ring-shaped marker 34 protruding from the chip body 33 toward the outer peripheral side is provided. More preferably, the lower surface (or upper surface) of the marker 34 may be orthogonal to the central axis of the chip body 33 in the longitudinal direction. Thereby, it is also possible to perform inclination correction based on the lower surface (or upper surface) of the marker 34 in image processing to be described later. More preferably, the distance between the tip surface of the chip 32 and the lower surface (or upper surface) of the marker 34 is defined to be a predetermined length H. This makes it possible to accurately grasp the position of the tip surface of the chip 32 in the image showing the chip 32.

  Referring again to FIG. 1, the dispensing pipette 3 has a moving mechanism 34 that holds the dispensing pipette 3 and operates under the control of the information processing device 6 to move the dispensing pipette 3 three-dimensionally. Is provided. The moving mechanism 34 moves the dispensing pipette 3 two-dimensionally along a plane (horizontal plane) parallel to the arrangement surface of the microplate 2, thereby positioning the dispensing pipette 3 with respect to the well 21 to be spotted. At the same time, the tip of the tip 32 is inserted into the well 21 by moving the dispensing pipette 3 in the vertical direction.

  In addition, the dispensing pipette 3 is provided with a drive mechanism 35 such as an electric motor that operates under the control of the information processing device 6 and drives the piston of the dispensing pipette 3. By controlling the driving amount of the piston by the driving mechanism 35, the amount of liquid sucked into the dispensing pipette 3 or the amount of liquid discharged from the dispensing pipette 3 can be automatically adjusted.

  The communication method between the moving mechanism 34 and the drive mechanism 35 and the information processing device 6 may be a wired method, or wireless using a wireless LAN, Wi-Fi (Wireless Fidelity), Bluetooth (registered trademark), or the like. It may be a method.

  Although only one dispensing pipette 3 is shown in FIG. 1, a single channel pipette may be used as the dispensing pipette 3, or a multichannel pipette in which a plurality of dispensing pipettes are arranged. It may be used. In this embodiment, a case where a single channel pipette is used will be described as an example. However, when a multichannel pipette is used, the same processing as that for a single channel pipette is performed on each pipetting pipette simultaneously. Just do it.

  The moving mechanism 34 described above is not essential, and the operator may move the dispensing pipette 3 manually. In this case, the operation of the drive mechanism 35 may be started with an operation on an operation switch provided in the dispensing pipette 3 as a trigger. In this case, a detection signal for detecting the operation of the drive mechanism 35 is wirelessly transmitted from the pipetting pipette 3 to the information processing device 6 so that it is possible to grasp that the spotting has been performed in the information processing device 6. May be.

  The camera 4 includes an image sensor 4a such as a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS), and can capture a still image or a moving image at a predetermined imaging frame rate (for example, 15 fps or more). Device. The image sensor 4a receives light (subject image) incident on the camera 4 and imaged by the optical system on the light receiving surface, and generates an electrical signal by performing photoelectric conversion. The camera 4 generates and outputs image data by performing predetermined signal processing such as amplification and A / D conversion on the electrical signal.

  The camera 4 may be a monochrome camera or a camera capable of generating a color image in which a color filter is disposed on the light receiving surface of the image sensor 4a. More preferably, a small camera such as a USB camera that can be connected to the information processing apparatus 6 via a USB (Universal Serial Bus) cable may be used so that the entire system does not become too large.

  As shown in FIG. 2, the camera 4 is installed so that the area above the microplate 2 is in the visual field V. More specifically, when the dispensing pipette 3 moves above the well 21 to be spotted and the liquid 100 is discharged, at least the tip portion of the tip 32 and the liquid 100 depending from the tip portion are stored in the visual field V. Thus, the camera 4 is installed. In the case where a marker 34 is provided on the chip 32, it is preferable that this marker 34 is also included in the field of view V.

  Regarding the visual field range of the camera 4 in the horizontal direction, the camera 4 may be installed so that the entire upper part of the microplate 2 is always in the visual field, or only the upper part of the well 21 to be spotted and its peripheral region are viewed. You may install the camera 4 so that it may house. In the latter case, a drive mechanism that changes the orientation of the camera 4 under the control of the information processing device 6 may be provided to change the field of view of the camera 4 in conjunction with the position of the dispensing pipette 3.

  Here, when the movement mechanism 34 is provided in the dispensing pipette 3 and the operations of the movement mechanism 34 and the drive mechanism 35 are controlled in conjunction with the camera 4 by the information processing device 6 as will be described later, the camera 4 is a still image or a comparison. A moving image may be captured at a relatively low frame rate (for example, 15 fps). On the other hand, when the operator manually operates the dispensing pipette 3, it is preferable to capture a moving image at a relatively high frame rate (for example, 60 fps or more).

  Preferably, the camera 4 is provided with a telecentric lens. Here, when the dispensing pipette 3 moves in the depth direction of the camera 4 above the microplate 2, when light is incident on the image sensor 4a through a normal condenser lens, the light receiving surface of the image sensor 4a The size of the image of the chip 32 in FIG. On the other hand, when light is incident on the image pickup device 4a via the telecentric lens, the size of the image of the chip 32 on the light receiving surface is kept within a certain range regardless of the distance between the chip 32 and the camera 4. be able to.

  The illumination device 5 is a light source that illuminates the field of view V of the camera 4, that is, the region including the tip of the chip 32. As the illumination device 5, it is preferable to use a surface light source having a planar light emitting surface so that the entire region above the microplate 2 can be illuminated with substantially uniform illuminance. More preferably, a light control film or the like that reduces the illumination light distribution angle is provided on the illumination light emitting surface in order to clarify the liquid contour. As an example, the illumination device 5 includes a flat illumination (for example, TH2-27X27BL manufactured by CCS Co., Ltd.) and a control film (for example, LC-TH-27X27-HO, LC-TH-27X27-HO manufactured by CCS Co., Ltd.). Combinations can be used.

  FIG. 3 is a block diagram illustrating a schematic configuration of the information processing apparatus 6. The information processing apparatus 6 is configured by, for example, a general-purpose device such as a personal computer (PC) or a notebook PC or a dedicated computer, and by spotting the microplate 2 by the dispensing pipette 3 by executing a predetermined program. While controlling the operation, in cooperation with the camera 4, various functions for measuring the amount of spotting deposited on each well 21 are realized.

  As illustrated in FIG. 3, the information processing apparatus 6 includes an image acquisition unit 61, a display unit 62, an input unit 63, a communication unit 64, a storage unit 65, and a calculation unit 66. The image acquisition unit 61 is an external interface that inputs and outputs image data and various signals to and from various external devices such as the camera 4. Specifically, a USB interface and the like are included.

The display unit 62 is, for example, a liquid crystal display or an organic EL display, and displays a predetermined screen under the control of the calculation unit 66.
The input unit 63 includes an input device such as a keyboard, a mouse, and a touch panel, and inputs information corresponding to an operation performed by the user to the calculation unit 66.

  The communication unit 64 is a communication interface that connects the information processing apparatus 6 to a communication network and transmits / receives information to / from other devices via the communication network. Specifically, an optical modem, a soft modem, a cable modem, etc. are included. The communication network may be, for example, WiFi, Bluetooth (registered trademark), the Internet, a LAN, a telephone line, or the like.

  The storage unit 65 is configured using a computer-readable storage medium such as a semiconductor memory such as a disk drive, ROM, or RAM. In addition to the operating system program and the driver program, the storage unit 65 stores a program for causing the information processing apparatus 6 to execute a predetermined operation, various data used during the execution of the program, setting information, and the like. Specifically, the storage unit 65 includes a program storage unit 651, a setting information storage unit 652, an image storage unit 653, and a spotting amount storage unit 654.

  The program storage unit 651 moves the dispensing pipette 3 by the moving mechanism 34 and drops the dispensing pipette 3 by the driving mechanism 35 in order to spot a predetermined amount of liquid in a predetermined order on the plurality of wells 21 of the microplate 2. The arithmetic unit 66 executes a spotting control program for causing the arithmetic unit 66 to execute control for discharging the liquid from the liquid and an arithmetic unit for measuring the amount of spotting to each well 21 based on the image acquired by the camera 4. A spot landing amount measurement program to be executed is stored.

The setting information storage unit 652 stores various setting information when dispensing. The setting information includes information such as position information of each well 21, spotting order, and the amount of spotting to each well 21.
The image storage unit 653 stores an image generated based on the image data output from the camera 4.
The spotting amount storage unit 654 stores the spotting amount to each well 21 calculated by the calculation unit 66.

  The calculation unit 66 is configured by using hardware such as a CPU (Central Processing Unit) and a GPU (Graphics Processing Unit), and reads and executes a program stored in the program storage unit 651, whereby the information processing apparatus 6 The data transfer to each unit and instructions are performed, and the operation of the information processing apparatus 6 is comprehensively controlled. Further, the calculation unit 66 controls the moving mechanism 34 and the driving mechanism 35 in order to spot a predetermined amount of liquid from the dispensing pipette 3 on the well 21 in a predetermined order by executing a spotting control program. Generate and output a control signal. Further, the calculation unit 66 generates an image based on the image data acquired from the camera 4 by executing the spotting amount measurement program, and points from the dispensing pipette 3 to each well 21 based on these images. An arithmetic process for calculating the amount of liquid applied is executed. Specifically, the calculation unit 66 includes an imaging control unit 661, a position control unit 662, a drive control unit 663, an image processing unit 664, and a spotting amount calculation unit 665.

The imaging control unit 661 generates a control signal for causing the camera 4 to perform imaging at a predetermined timing in accordance with various setting information stored in the setting information storage unit 652.
The position control unit 662 generates a control signal for controlling the operation of the moving mechanism 34 in accordance with the various setting information stored in the setting information storage unit 652.
The drive control unit 663 generates a control signal for controlling the operation of the drive mechanism 35 in accordance with various setting information stored in the setting information storage unit 652.

The image processing unit 664 generates an image by performing predetermined image processing such as demosaicing, white balance processing, and gamma correction on the image data output from the camera 4 and acquired through the image acquisition unit 61. At the same time, predetermined image processing such as difference image generation and binarization processing is executed on the generated image.
The spotting amount calculation unit 665 executes a spotting amount calculation process based on the image generated by the image processing unit 664.

  In the present embodiment, the control for the moving mechanism 34 and the driving mechanism 35 and the image processing for the image data output from the camera 4 and the calculation processing of the spotting amount are performed in one device (information processing apparatus 6). However, the control of the moving mechanism 34 and the driving mechanism 35, the image processing, and the calculation of the amount of spotting may be performed in separate devices. Also in the latter, by performing communication between these devices, the spotting operation of the dispensing pipette 3 and the imaging operation for spotting amount measurement can be executed in cooperation with each other.

Next, the operation of the spotting amount measurement system 1 will be described with reference to FIGS. FIG. 4 is a flowchart showing the operation of the spotting amount measurement system 1.
Prior to the start of spotting, in step S10, the information processing apparatus 6 acquires a reference image that is an image in which only the tip portion of the chip 32 is shown. That is, as shown in FIG. 5, imaging is executed by aligning the field of view V of the camera 4 with the tip of the chip 32, and an image is generated based on the image data output from the camera 4. At this time, if the chip 34 is provided with the marker 34, the marker 34 is also stored in the field of view V. After that, spotting starts.

In subsequent step S <b> 11, the position controller 662 moves the dispensing pipette 3 onto the well 21 to be spotted by operating the moving mechanism 34.
In subsequent step S <b> 12, the drive control unit 663 operates the drive mechanism 35 to discharge the liquid from the dispensing pipette 3 and hang down the tip of the tip 32.

  In subsequent step S <b> 13, the information processing apparatus 6 acquires an image before spotting that is an image showing a state before spotting on the well 21 by the dispensing pipette 3. That is, as illustrated in FIG. 6, the imaging control unit 661 causes the camera 4 to image the tip of the chip 32 in the field of view V and the liquid 100 depending on the tip 32, and the image processing unit 664 is output from the camera 4. An image before spotting is generated based on the image data.

  In subsequent step S14, the position controller 662 operates the moving mechanism 34 to move the dispensing pipette 3 downward, and inserts the tip end of the tip 32 into the well 21 as shown in FIG. Let it be spotted. Thereafter, the position controller 662 operates the moving mechanism 34 again, and pulls up the dispensing pipette 3 to the same position as before the spotting (see step S12).

  In subsequent step S <b> 15, the information processing apparatus 6 acquires a post-dotting image that is an image showing a state after spotting on the well 21 by the dispensing pipette 3. That is, as illustrated in FIG. 8, the imaging control unit 661 causes the camera 4 to image the tip of the chip 32 in the field of view V, and the image processing unit 664 displays the image after spotting based on the output image data. Generate.

  In subsequent step S16, the image processing unit 664 performs predetermined image processing on the acquired image. Specifically, the image processing unit 664 aligns and corrects the inclination between the pre-potting image in which the subject in the visual field V shown in FIG. 6 is captured and the reference image in which the subject in the visual field V shown in FIG. Then, a difference image M1 between the two images (see FIG. 9) is generated. Further, the binarization process is performed on the difference image M1, thereby extracting the liquid region m10 that appears in the image before spotting. Further, the image processing unit 664 performs alignment and inclination correction between the image after spotting in which the subject in the field of view V shown in FIG. 8 is captured and the reference image, and then the difference image M2 between the two images. (See FIG. 9). Further, the binarization process is performed on the difference image M2, thereby extracting the liquid region m11 shown in the post-dotting image.

  In subsequent step S17, the spotting amount calculation unit 665, based on the area m10 extracted from the difference image M1 from the image before spotting and the area m11 extracted from the difference image M2 from the image after spotting, The amount of spotting is calculated. Specifically, first, the area m10 is regarded as a projected image of the liquid depending from the tip of the chip 32, and the volume of the original liquid is calculated. Further, the area m11 is regarded as a projected image of the liquid remaining on the chip 32, and the volume of the original liquid is calculated. The method for calculating the volume of these liquids is not particularly limited. For example, the volume of the rotating body obtained by rotating each of the areas m10 and m11 around the vertical axis may be obtained by integration calculation. Or you may use the method of slicing each of the area | regions m10 and m11 by the line orthogonal to a vertical axis, and integrating | accumulating the volume of the disk which makes the width | variety of each fragment | piece the diameter. In calculating the volume, the sizes (pixels) of the regions m10 and m11 in the image are converted to the corresponding sizes in the actual liquid based on the width D (see FIG. 2) of the tip of the chip 32 and the like.

  Then, the spotting amount calculation unit 665 subtracts the volume of the liquid remaining at the tip portion of the chip 32 from the volume of the liquid dripping from the tip portion of the chip 32. As shown in FIG. 9, this corresponds to the volume of the object m12 obtained by hollowing out a part of the base end portion from the liquid hanging from the tip end portion of the chip 32. The spotting amount calculation unit 665 outputs the volume of the object m12 calculated in this way as the volume of the liquid that is substantially spotted in the well 21.

  Thereafter, when an unattached well 21 remains (step S18: No), the operation of the spotted amount measuring system 1 returns to step S11. On the other hand, when the spotting to all the set wells 21 is completed (step S18: Yes), the operation of the spotting amount measurement system 1 is terminated.

  As described above, according to the present embodiment, the amount of liquid remaining at the tip of the chip 32 in the image after spotting is determined based on the volume of the liquid hanging from the tip of the chip 32 in the image before spotting. Since the amount of spotting is calculated by subtracting the volume, it is possible to accurately grasp the amount of liquid that is substantially spotted on the well 21. Therefore, it is possible to improve the accuracy of the inspection or experiment by performing the inspection or experiment based on the spotting amount calculated in this way.

  Further, according to the present embodiment, by providing the marker 34 on the chip 32, it is possible to accurately perform image inclination correction and alignment between images based on the image of the marker 34 shown in each image. It becomes possible. Here, since the chip is generally formed of a translucent member, it is difficult to identify the boundary between the chip and the liquid dripping from the tip thereof (that is, the end face of the chip), and the end face of the chip is used as a reference. In addition, it is difficult to perform image tilt correction and alignment. In particular, in the present embodiment, as shown in FIG. 1, since the chip 32 is illuminated from behind by the illumination device 5, the image of the chip 32 is dark in the image captured by the camera 4. Therefore, it is very difficult to detect the end face of the chip. Therefore, by providing a marker 34 that protrudes toward the outer peripheral side of the chip 32, it becomes possible to clearly identify a reference for tilt correction and alignment (the lower surface of the marker 34).

  Of course, the marker provided on the chip 32 is not limited to the marker 34 protruding from the chip body 33 toward the outer peripheral side as shown in FIG. For example, a color line or band as a marker may be drawn continuously or intermittently along the circumference on the outer peripheral surface of the chip body 33. Alternatively, a line may be imprinted on the outer peripheral surface of the chip body 33 as a marker, or a step may be provided. In these cases, the marker is clearly identified by arranging the illumination device so that the front side of the chip 32 (the side facing the camera 4) is also illuminated and using the camera 4 capable of acquiring a color image. Can do.

  In this way, by using the tip 32 provided with the marker 34 in the spotting amount measurement system 1, it is possible to improve the extraction accuracy of the liquid areas m10 and m11 in the image before spotting and the image after spotting. It is possible to ensure the measurement accuracy of the amount of wearing, and hence the accuracy of inspections and experiments. That is, the marker 34 provided on the chip 32 can be used as a precision guarantee.

  In addition, according to the present embodiment, the imaging control unit 661, the position control unit 662, and the drive control unit 663 control the imaging operation of the camera 4 in conjunction with the movement of the dispensing pipette 3 and the operation of discharging the liquid. Therefore, it is possible to generate an image before spotting and an image after spotting in which the tip end portion of the chip 32 is securely stored in the field of view V of the camera 4.

Next, a modified example of the spotting amount measurement method according to the present embodiment will be described. FIG. 10 is a schematic diagram for explaining a modification of the spotting amount measuring method.
The calculation of the spotting amount may be performed using only the pre-spotting image and the post-spotting image without using the reference image. That is, as shown in FIG. 10, a difference image is generated by subtracting the image M4 after spotting from the image M3 before spotting, and a binarization process is performed on the difference image to extract the region m13. To do. Prior to the generation of the difference image, it is preferable to perform alignment between the image before spotting and the image after spotting based on the image of the marker 34. Then, the volume of the rotating body obtained by rotating the region m13 about the vertical axis is set as a substantial amount of spotting spotted in the well 21. The volume calculation method is the same as in the above-described embodiment.

  According to this modification, after the start of spotting, unnecessary liquid or the like adheres to the chip 32 or the like, and only the liquid region is extracted from the difference image between the image before spotting or the image after spotting and the reference image. Even when it becomes difficult, it is possible to accurately calculate the amount of spotting.

  The present invention is not limited to the embodiments described above, and can be implemented in various other forms without departing from the gist of the present invention.

DESCRIPTION OF SYMBOLS 1 Spot amount measurement system 2 Microplate 3 Dispensing pipette 4 Camera 4a Image pick-up element 5 Illumination device 6 Information processing apparatus 21 Well 31 Pipette main body 32 Tip 33 Tip main body 34 Marker 34 Movement mechanism 35 Drive mechanism 61 Image acquisition part 62 Display part 63 Input unit 64 Communication unit 65 Storage unit 66 Calculation unit 100 Liquid 651 Program storage unit 652 Setting information storage unit 653 Image storage unit 654 Spot amount storage unit 661 Imaging control unit 662 Position control unit 663 Drive control unit 664 Image processing unit 665 Spot amount calculation unit

Claims (11)

An imaging device that images a subject and outputs image data;
Based on the image data output from the imaging device, the liquid is imaged before being spotted on the spotted material by the dispensing pipette, and at least the tip portion of the tip provided on the dispensing pipette and the liquid depending from the tip portion And an image before spotting that is an image in which the tip is spotted by the pipette pipette and at least the tip portion of the tip is captured. An information processing device that calculates a spotting amount that is a volume of liquid spotted on the spotted material based on the pre-spotting image and the post-spotting image;
A spot amount measuring system comprising: The information processing apparatus includes:
A reference image that is an image in which the tip portion of the chip in a state in which no liquid is drooping is captured based on the image data output from the imaging device while generating the pre-drop image and the post-drop image. Generating a first difference image between the pre-spotting image and the reference image, extracting a first region in which the liquid appears from the first difference image, and the post-spotting image; An image processing unit that generates a second difference image from the reference image and extracts a second region in which the liquid is reflected from the second difference image;
A spotting amount calculation unit that calculates the spotting amount based on the extracted first and second regions;
The spot amount measuring system according to claim 1, comprising:   The image processing unit further performs alignment based on the image of the chip between the image before spotting and the reference image and between the image after spotting and the reference image. Item 3. The spot amount measuring system according to Item 2. The information processing apparatus includes:
Image processing for generating the image before spotting and the image after spotting, generating a difference image between the image before spotting and the image after spotting, and extracting an area in which the liquid appears from the difference image And
A spotting amount calculation unit that calculates the spotting amount based on the extracted region;
The spot amount measuring system according to claim 1, comprising:   The spotting amount measurement system according to claim 4, wherein the image processing unit further performs positioning based on the image of the chip between the pre-spotting image and the post-spotting image. The chip is
A chip body for discharging liquid;
A marker provided on the outer peripheral surface of the chip body;
Have
The imaging device is arranged so that at least the marker is in the field of view,
The spotting amount measurement system according to claim 3 or 5, wherein the image processing unit performs the alignment based on an image of the marker.   7. The spotting amount measurement system according to claim 6, wherein the image processing unit further performs tilt correction of the pre-dotting image and the post-spotting image based on the marker image.   The spotting amount measurement system according to claim 1, wherein the imaging device includes a telecentric lens.   The spotting amount measurement system according to claim 1, further comprising a light source that illuminates at least a field of view of the imaging device. Based on image data acquired by imaging before spotting a spotted material with a dispensing pipette, at least the tip of the tip provided in the dispensing pipette and the liquid dripping from the tip Generating a pre-dotted image that is a captured image;
Based on image data acquired by performing imaging after spotting on the spotted material with the pipetting pipette, generating a post-spotting image that is an image showing at least the tip of the tip;
Calculating a spotting amount that is a volume of liquid spotted on the spotted material based on the pre-spotting image and the post-spotting image;
A method for measuring the amount of spotting including In a spotting amount measurement system comprising: an imaging device that captures an image of a subject and outputs image data; and an information processing device that performs information processing based on image data output from the imaging device. A program,
Before the spotting material is spotted by the dispensing pipette, the imaging device executes imaging, and based on the image data output from the imaging device, at least the tip of the tip provided in the dispensing pipette and the tip Generating a pre-dotted image that is an image of liquid dripping from the tip;
After spotting the spotted material with the pipetting pipette, the imaging apparatus causes the imaging apparatus to perform imaging, and based on image data output from the imaging apparatus, the spotting is an image in which at least the tip of the tip is reflected Generating a post-image;
Calculating a spotting amount that is a volume of liquid spotted on the spotted material based on the pre-spotting image and the post-spotting image;
A program that executes

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