JP2000206057A - Verification device of fine object accommodated in container - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a device for accurately and efficiently verifying a fine object being accommodated in a container. SOLUTION: A verification device is provided with a storage means for relating data on a fine object being sampled from a liquid to information on a micro plate where the fine object is accommodated for storage, a display monitor 85 for displaying the graphic image of the micro plate and the data of the fine object, a display processing means 96 for mutually relating the graphic image and the data of the fine object and displaying them on the same screen of above mentioned display device, and an input part 86 for giving an instruction to the display means, thus mutually relating the graphic image of the micro plate and the data on the fine object being accommodated and displaying on the same screen and hence accurately and efficiently verifying the fine object after sampling.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a device for checking a fine object contained in a container such as a microplate.

[0002]

2. Description of the Related Art In the process of various operations in the field of biochemistry and the like, an operation of searching for, identifying, and collecting a specific fine object from a large number of fine objects present in a liquid sample is performed. Conventionally, a manual method has been generally used in which a skilled expert observes and identifies a fine object within a visual field of a microscope, and collects the identified fine object by operating a micropipette. Such a method relying on manual work is extremely inefficient, and the identification result is inaccurate and unreliable. For this reason, in recent years, a sampling device that can more easily collect the identified fine object has been used. A large number of fine objects collected by this collection device are collectively stored in a container such as a microplate. Thereafter, these fine objects are subjected to various operations such as analysis and culturing, and in order to perform these operations systematically to obtain correct test results, strict separation management of the collected fine objects is required.

[0003]

However, since fine objects are invisible to the naked eye, there has been a great deal of difficulty in sorting and managing the fine objects after being housed in the microplate. In other words, a method for efficiently and accurately confirming what kind of fine object is contained in which well of which microplate has not been established. There is a problem that it takes a lot of time and effort to confirm the condition.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a fine object confirming apparatus capable of accurately and efficiently confirming a fine object contained in a container.

[0005]

SUMMARY OF THE INVENTION According to the present invention, a device for confirming a fine object contained in a container is provided by associating data on the fine object collected from the liquid with information on the container containing the fine object. A storage device for storing, a display device for displaying the graphic image of the container and the data of the fine object, and a display processing device for displaying the graphic image and the data of the fine object on the same screen on the same display device in association with each other. Instruction means for giving an instruction to the display processing means.

According to the present invention, the graphic image of the container and the data of the contained fine object are displayed on the same screen in association with each other, whereby the fine object can be confirmed accurately and efficiently.

[0007]

(Embodiment 1) FIG. 1 is a side view of an apparatus for automatically collecting a fine object in which a device for confirming a fine object according to a first embodiment of the present invention is incorporated, and FIG. FIG. 3 is a block diagram showing the configuration of the automatic sampling apparatus, and FIG. 3 is a functional block diagram showing the processing functions of the automatic sampling apparatus for fine objects.
(A), (b), FIG. 5 is a view showing a display screen of the apparatus for automatically collecting fine objects, and FIGS. 6 and 7 are flowcharts of a method for collecting the fine objects by the automatic collection apparatus.

First, the structure of an automatic fine object sampling apparatus will be described with reference to FIG. In FIG. 1, an automatic sampling apparatus 1 for a fine object is configured by arranging respective components described below inside a cover 2. The cover 2 is made of a heat-insulating member and includes an environment controller 3 that is an environmental condition holding unit that holds environmental conditions of the internal environment of the cover 2. By driving the environment controller 3, the temperature and temperature of the environment in which the minute object in the sample accommodated in the cover 2 is placed
Environmental conditions such as humidity and carbon dioxide concentration can be kept constant.

A base 5 is provided inside the cover 2. A side frame 5a is provided upright at an end of the base 5, and an upper frame 5b is provided above the side frame 5a.
Is fixed. A first XY table 6 is provided on a side surface of the side frame 5a, and a sample table 7 having a light-transmitting property is mounted on the first XY table 6. The sample table 7 supports a translucent sample container 8 containing a liquid sample containing a microscopic object such as a microorganism, animal or plant cell.

The upper frame 5b is provided with a transmitted light 9 (first illuminator).
The sample container 8 is irradiated with the illumination light reflected by. A camera 18 is provided at the bottom of the base 5 below the sample container 8. A fluorescent lamp 10 (second lighting unit) is provided on the side of the base 5, and the fluorescent lamp 10 is a half mirror 1.
Illumination light is irradiated from below the sample container 8 via the reference numeral 5. The camera 18 captures an image of the sample container 8 illuminated by the transmitted light 9 or the fluorescent light 10.

An infrared laser light source 11 is provided on the side of the base 5.
(First laser device) and an ultraviolet laser light source 12 (second laser device) are provided. The infrared laser light emitted from the infrared laser light source 11 is irradiated upward by the mirror 17 via the scanning optical system 13 and enters the sample container 8 from below. By irradiating a fine object in the sample liquid of the sample container 8 with infrared laser light, the fine object is trapped and captured by the optical pressure of the infrared laser light.
By causing the scanning optical system 13 to scan the infrared laser light, the fine object to be captured in the sample container 8 can be irradiated with the infrared laser light, trapped, and moved to an arbitrary position without contact. . Infrared laser light source 1
The scanning optical system 1 and the scanning optical system 13 are so-called laser tweezers for trapping a fine object by a laser beam, and serve as a capturing unit for capturing the fine object.

The ultraviolet laser light emitted from the ultraviolet laser light source 12 is reflected by the mirror 16 and is
From below. This ultraviolet laser light is suitable for a processing application in which a fine object in a sample is locally destroyed, and processing such as cutting of the fine object can be performed by irradiating the ultraviolet laser light to the fine object to be processed. The ultraviolet laser light source 12 is a processing means.

Above the sample container 8, a sample supply pipette 21 is guided, and the sample supply pipette 21 is XYZ.
It can be moved by the table 22. The sample supply pipette 21 is connected to a dispensing mechanism 25 to which a sample tank 29, a diluent tank 30 for cleaning, and a waste liquid tank 31 are connected via pipes 26, 27, 28. I have. By using the suction / discharge mechanism of the dispensing mechanism 25, the sample and the diluent sucked from the sample tank 29 and the diluent tank 30 are selectively discharged from the tip of the sample supply pipette 21 to the sample container 8. The extra sample remaining in the sample container 8 after the test and the diluent used for washing are sucked by the dispensing mechanism 25 and collected in the waste liquid tank 31.

An XYZ table 2 is provided near the sample container 8.
Micropipette for collection 23 movable by 4
However, it is arranged in such a manner that the suction port at the tip is immersed in the liquid of the sample container 8. The micropipette 23 is connected to the dispensing mechanism 25, and by using the suction / discharge function of the dispensing mechanism 25, a microscopic object contained in the sample in the sample container 8 and located near the suction port of the micropipette 23 is removed. It can be collected by inhalation and discharged from the tip of the dispensing nozzle 36. Therefore, the micropipette 23 is a sampling unit for sampling a fine object together with the dispensing mechanism 25.

Below the dispensing mechanism 25, a second XY table 32 is provided. A plate holder 33 is mounted on the second XY table 32, and a microplate 34 having a well 35, which is a container for a plurality of fine objects, is mounted on the upper surface of the plate holder 33. By driving the second XY table 32, the dispensing nozzle 3
By horizontally moving the microplate 34 located immediately below 6, the fine object discharged from the dispensing nozzle 36 can be injected into an arbitrary well 35.

Next, the configuration of the control system of the automatic sampling apparatus will be described with reference to FIG. The camera 18, the first illumination unit, and the second illumination unit 10 capture an image of a fine object in the sample container 8 according to an instruction from the CPU 43. The dispensing controller 41
The operation of the dispensing mechanism 25 is controlled. The scanning optical system controller 42 moves the trapping infrared laser light within the sample container 8 by controlling the scanning optical system 13. C
The PU 43 includes a storage unit such as a RAM 44, a ROM 45,
Various processes and calculations are executed based on programs and data stored in the file device 46. RAM 44 is CP
This is a memory for temporarily storing data during the processing operation of U43. The ROM 45 stores programs necessary for various processes and operations.

The file device 46 stores programs and various data, for example, images of fine objects captured by the camera 18 and classification results of these fine objects. The storage medium drive 47 writes and reads necessary programs and data to and from an external storage medium. The external storage medium 48 is a storage medium such as a floppy disk,
Used for data storage. The display monitor 49 is a display, and displays a screen at the time of operation and input, and displays an image of a captured fine object. The input unit 50 is a keyboard or a pointing device, and inputs operation commands and data. In addition, a touch panel, a joystick, or the like may be used as the instruction means.

Next, the processing function of the automatic sampling apparatus will be described with reference to FIGS. In FIG. 3, an image processing unit 51, a capturing operation control unit 52, a sampling operation control unit 5
3. The classification processing means 54 and the display processing means 55
3 is processing means (processing function). Further, each storage unit described below stores RA as storage means.
The content stored in the M44, the ROM 45, and the file device 46 is shown.

The image processing means 51 recognizes a target fine object from the image of the fine object in the sample container 8 captured by the camera 18 by image processing. Therefore, the image processing means 51 is a search means for searching for a target fine object in the sample. The reference image storage unit 56 stores a reference image used as a search condition. The search condition storage unit 57 stores search conditions for performing a search process for specifying a target fine object from an image of the fine object in the field of view obtained by the image processing.

The collected fine object image storage section 58 stores an image of the searched and collected fine object. The microplate information storage unit 59 stores information such as the arrangement of the wells of the microplate that stores the collected fine objects. The collection result storage unit 60 stores data relating to the image, type, and the like of the collected fine object in association with information on a well in which the fine object is stored. As data relating to the fine object, there is data (color, size, shape, etc.) indicating the characteristics of the fine object obtained by the image processing means 51.

The display processing means 55 displays on the display monitor 49 a device operation from the input unit 50, an operation screen at the time of data input, and an input screen. Information on the microplate containing the object is displayed on the display monitor 49.

Next, the contents of the above processing functions will be described. Image data of the sample imaged by the camera 18 is subjected to image processing by the image processing means 51. Then, it is determined whether or not the image of each fine object in the field of view obtained by this image processing matches the search condition for specifying the target fine object, based on the data in the reference image storage unit 56 and the search condition storage unit 57. They are compared and judged. The search conditions used here include those based on the degree of coincidence with the image stored as the reference image, those based on the size, shape, and color,
Further, various search conditions that can be set based on image information, such as conditions set based on a movement pattern such as a moving speed and a trajectory of a fine object in a visual field, are set.

By irradiating an infrared laser beam of the first laser device 11 to a fine object searched for and specified as a target fine object by the above processing, the fine object is trapped by light pressure. The capturing operation by this trap is performed by the capturing operation control means 52 by the scanning optical system controller 42, the first laser device 11, and the second laser device 1.
2. It is performed by controlling the first XY table 6.

The reference image of the fine object used in the search processing is stored in a reference image storage unit 56 as storage means. Data such as a size, shape, color, a movement pattern such as a trajectory and a moving speed used as a search condition is input from the input unit 50 and stored in the search condition storage unit 57. In addition, the image data of the fine object that has been captured and collected as a result of the search is stored in the collected fine object image storage unit 58.

When a fine object searched for by image processing is collected, if there are a plurality of types of fine objects, a classification process is performed assuming classification management after the collection.
That is, for a plurality of microscopic objects to be searched and collected from the same sample based on a plurality of search conditions, a determination is made as to which microplate, which well accommodates the microscopic object corresponding to the search condition in which well of the microplate. . This decision
Based on the information on the microplate to be used stored in the microplate information storage unit 59, the classification processing unit 5
4 done.

Then, based on this determination, the searched and collected fine object is transferred from the sample container 8 to the micropipette 2.
3, a process of dispensing the collected fine object to the microplate 34, that is, a dispensing process of storing the collected fine object in the designated well 35 of the microplate 34 is performed. These processes are performed by controlling the second XY table 32, the XYZ table 24, and the dispensing controller 42 by the sampling operation control means 53. That is, the microplate 34 is a classification unit that classifies the fine object based on the search condition.
The well 35 of the microplate 34 is a container for storing a fine object.

After the search / collection and the above-described dispensing process, the information on the collected fine object and the well of the microplate in which the fine object is stored is stored in the collection result storage unit 60. Based on the information, a data screen as shown in FIGS.
5 is created.

The screen shown in FIG. 4A displays what kind of minute object is contained in which well of which microplate. In the example shown in FIG. 4A, by inputting the plate number and the well position into the display boxes 70 and 71 on the screen, the position of the well is illustrated on the graphic image 72 of the microplate and stored in the well. The name of the fine object, the characteristic data obtained by the image processing, and the image 73 are displayed. Instead of inputting a number as a method of designating a well, the cursor 77 on the screen can be operated from the input unit 50 to designate the well represented by the graphic image 72. In this case, when the well is designated on the image, the plate number and the well position are displayed in the display box 7.
0, 71, the name and characteristic data of the fine object contained in the well and the image 73 are displayed.
When data for specifying the fine object is input to the display boxes 73a, 73b, and 73c such as the name, feature, and image number of the stored fine object from the input unit 50, the position of the well containing the corresponding fine object is displayed as a graphic image. 7
2 shown above.

As described above, by designating the position of the well of the microplate and any one of the fine objects contained in the well, the corresponding relationship mutually associated is visually and clearly displayed on the same screen. You. This display method includes displaying the well in a different color from the other wells as shown in FIG. 4A, blinking the display of the well, or as shown in FIG. 4B. For example, a method of displaying a cross cursor 78 on the well can be used. FIG. 5 shows a collection result list, in which the microplate number 74, the coordinate number 75 of the well position, and the registration number 76 added to the image are displayed for each serial number given to the collection operation.

In the work of confirming a fine object after collection performed using the screen, all necessary instructions can be performed using the input unit 50 on the screen of the display monitor 49. That is, necessary instructions are performed while visually confirming the screen by a keyboard input, a cursor instruction by a mouse, or the like. Therefore, the input unit 50 is an instruction unit for giving an instruction to the display processing unit 55.

As described above, information such as the name, characteristic data, and image of the fine object is stored in the collection result storage unit 60 as storage means in association with information on the well in which the fine object is stored. . Then, in the work of confirming the fine object after collection, necessary data is read from the information and displayed by the display processing means 55, and the graphic image of the microplate and the information of the fine object are associated with each other and displayed on the display device. Are displayed on the same screen on the display monitor 49.

Next, a method of collecting a fine object by the automatic collecting apparatus will be described with reference to the drawings and along the flow charts of FIGS. In FIG. 1, a diluting liquid is first sucked from a diluting liquid tank 30 by a dispensing mechanism 25 and discharged from a sample supply pipette 21 to wash the sample container 8 on the sample table 7 (ST1). Next, the sample including the fine object is sucked from the sample tank 29 by the dispensing mechanism 25, discharged from the sample supply pipette 21, and supplied into the sample container 8 (ST2).

Thereafter, the camera 18 captures an image of the sample containing the liquid in the sample container 8 (ST3), and performs a search process of recognizing and detecting a target fine object from the obtained image by image processing. (ST4). That is, a target fine object is identified from the sample. Here, if the target fine object is not found at all in the sample to be searched, the process returns to the first ST1, and the same processing is repeated for a new sample.

When the target fine object is found (ST5), the image data of the found fine object, that is, the fine object to be searched is collected by the sampled fine object image storage unit 58.
(ST6), and by irradiating an infrared laser beam, the identified target fine object is trapped and captured with optical pressure by laser tweezers, and transferred to the sampling position of the micropipette 23 (ST7).

Then, the position of the well accommodating the fine object is determined by the classification processing means 54 (ST8), and the second XY table 32 is driven so that the determined well is located immediately below the dispensing nozzle 36. Microplate 3
4 is positioned (ST9). Then XYZ table 2
4 is driven to move the tip of the micropipette 23 to the collection position (ST10), and the dispensing mechanism 25 is driven to aspirate and collect the captured target fine object with the micropipette 23, and then to a predetermined well. 35 and accommodated (ST
11).

At this time, the position of the housed well and the number of the image stored in ST6 are stored in the collection result storage unit 60 as the collection result data (ST12). That is, information on the collected fine object is stored in association with the well of the accommodation destination. Thereafter, it is determined whether or not a necessary number of fine objects have been collected (ST13). If not, the process returns to ST3 and the subsequent steps are repeated. Then, the sample table 7 is washed again (ST14), and the collection process is terminated.

The data obtained during the collection process and stored in the collected fine object image storage unit 58, the microplate information storage unit 59, and the collection result storage unit 60 are stored in an external storage such as a floppy disk 48 by the storage medium drive unit 47. Stored on the medium. These data are used at the time of a checking operation by a checking device provided separately from the sampling device as described in the second embodiment.

(Embodiment 2) FIG. 8 is a perspective view of a fine object confirmation apparatus according to a second embodiment of the present invention, FIG. 9 is a block diagram showing a configuration of the fine object confirmation apparatus, and FIG. FIG. 3 is a functional block diagram illustrating a processing function of the object confirmation device.
In the second embodiment, only the confirmation function for displaying the data of the collected fine object and the information on the microplate in which the fine object is accommodated is extracted.
This is a device for checking a single fine object.

Referring to FIG. 8, a device 80 for checking a fine object includes a personal computer 81. Data on the micro object and the micro plate containing the micro object separately acquired by the micro object collection device are stored in an external storage medium such as a floppy disk 82 and supplied to the present micro object confirmation device. The operator operates the mouse 83 and the keyboard 84 to display these data on the display monitor 85, and performs necessary instructions on this screen to confirm the collected fine object.

Next, the configuration of a device for checking a fine object will be described with reference to FIG. The display monitor 85 and the input unit 8 shown in FIG.
6, the CPU 87, the RAM 88, the ROM 89, the file device 90, and the storage medium drive device 91 are the display monitor 49, the input unit 50, and the CP in the first embodiment (see FIG. 2).
U43, RAM 44, ROM 45, file device 46,
It has basically the same function as the storage medium drive 47. However, in the first embodiment, the above-described units are targeted for processing including search and collection of a fine object, whereas in the second embodiment, only a fine object confirmation operation is performed based on acquired data. Embodiment 2 is different from Embodiment 1 in that it is limited to

Next, with reference to FIG. 10, the processing function of the confirmation difference of the fine object will be described. 10, a display processing means 96 is a processing function executed by the CPU 7, and a sampling result storage unit 93, a microplate information storage unit 94, and a collected fine object image storage unit 95 are a RAM 88, a ROM 89, and a file device as storage units. 90 shows the contents stored. The contents of the data stored in each storage unit are the same as the storage contents of the collection result storage unit 60, the microplate information storage unit 59, and the collected fine object image storage unit 58 in the first embodiment (see FIG. 3). .

As described above, the data stored in these storage units is read by the storage medium drive unit 91 using the floppy disk 82 as a medium and stored in each of the storage units. The information may be stored in the file device 90 in advance, or may be directly read from the floppy disk 82 during operation. Further, a configuration may be adopted in which information necessary for each storage unit is obtained by communication means without using a storage medium.

The display processing means 96 creates the data screens shown in FIGS. 4 and 5 in the first embodiment based on the data stored in the storage means, similarly to the display processing means 55 in the first embodiment, and It is displayed on the display monitor 85 according to the instruction input to the unit 86. As described above, in the second embodiment, data obtained when a fine object is collected by the fine object collecting device described in the first embodiment, that is, data on a micro object and a microplate containing the fine object. Is stored in a storage medium, and when a fine object as a result of collection is confirmed at a remote place separately from the collection device, a confirmation operation is efficiently performed based on the data.

As described in the first and second embodiments, the data such as the image of the collected fine object and the graphic image of the microplate containing the fine object are displayed on the same screen on the display monitor. By performing the checking operation while visually recognizing the mutual relationship, the checking operation of a fine object that cannot be visually recognized can be accurately and efficiently performed.

[0045]

According to the present invention, the graphic image of the container and the data of the fine object contained in the container are displayed on the same screen in association with each other.
It is possible to accurately and efficiently specify a fine object contained in a container and confirm a specified result after collection.

[Brief description of the drawings]

FIG. 1 is a side view of an automatic fine object sampling apparatus incorporating a fine object confirmation apparatus according to a first embodiment of the present invention;

FIG. 2 is a block diagram showing a configuration of an automatic fine object sampling apparatus according to the first embodiment of the present invention.

FIG. 3 is a functional block diagram showing processing functions of the automatic fine object sampling apparatus according to the first embodiment of the present invention;

FIG. 4A is a diagram showing a display screen of the apparatus for automatically collecting fine objects according to the first embodiment of the present invention; and FIG. 4B is a diagram showing a display screen of the apparatus for automatically collecting fine objects according to the first embodiment of the present invention.

FIG. 5 is a diagram showing a display screen of the apparatus for automatically collecting a fine object according to the first embodiment of the present invention.

FIG. 6 is a flowchart of a collection method using the automatic collection apparatus for a fine object according to the first embodiment of the present invention.

FIG. 7 is a flowchart of a collection method using the automatic collection apparatus for a fine object according to the first embodiment of the present invention;

FIG. 8 is a perspective view of a device for checking a fine object according to the second embodiment of the present invention;

FIG. 9 is a block diagram showing a configuration of a device for checking a fine object according to the second embodiment of the present invention;

FIG. 10 is a functional block diagram showing processing functions of the fine object checking device according to the second embodiment of the present invention;

[Explanation of symbols]

 8 Sample container 18 Camera 23 Micropipette 25 Dispensing mechanism 34 Microplate 35 Well 43 CPU 49, 85 Display monitor 50, 86 Input unit 58, 95 Collection fine object image storage unit 59, 94 Microplate information storage unit 60, 93 Collection Result storage unit

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G051 AA90 AB20 AC12 AC21 BA01 BA05 BA06 BA10 BB11 BC05 CA04 CB02 CC11 DA07 EA11 EA12 EA14 EB09 ED21 FA01

Claims (2)

[Claims] 1. A storage means for storing data on a fine object collected from a liquid and information on a container in which the fine object is stored, and displaying a graphic image of the container and data on the fine object. And a display processing means for associating the graphic image and the data of the fine object with each other and displaying the data on the same screen on the display device, and an instruction means for giving an instruction to the display processing means. Confirmation device for fine objects contained in a container. 2. The apparatus according to claim 1, wherein the data of the fine object includes at least an image of the fine object.