JP3465613B2 - Operating device for fine objects - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [0001] The present invention relates to the field of biochemistry and the like.
Microscopic objects such as microorganisms and animal and plant cells in the sample solution
Device for moving, capturing, and collecting fine objects
It is about. [0002] 2. Description of the Related Art Various tests and analyzes in the biochemical field, etc.
Is a microscopic object such as a large number of microorganisms present in a sample.
For searching and identifying specific fine objects from within
Is performed. Traditionally, these operations have been performed by skilled
Observe and identify minute objects within the microscope field of view,
Collect the identified fine object using a micropipette, etc.
The manual method of taking was commonly used. [0003] SUMMARY OF THE INVENTION
In operation, workers observe and identify fine objects,
Aside from the original work of identifying the target of collection
It was in a situation where a great deal of labor was required for the extra work. You
In other words, the identified fine object is
To capture and move the captured fine object to the microphone.
Workers who use the same pipette for sampling
Had to be performed, the work of collecting fine objects
Is extremely inefficient and time consuming.
There was a problem. Accordingly, the present invention makes it possible to efficiently collect fine objects.
To provide a device for manipulating fine objects
With the goal. [0005] [0006] [0007][Means for Solving the Problems] The present invention Manipulation of fine objects
The device captures an image of a fine object in the liquid stored in the container.
Camera and the image taken by this cameraAnd micro
Rate graphic images on the same screen at the same timedisplay
Display device and a micro-
A microplate support that supports the rate,The container
Irradiates laser light into the
Capture means for capturing fine objects in the body and transferring them to the sampling position
And transferred to the sampling locationSuction fine objects
Collection means for discharging into wells of a microplate;
A fine object on the image displayed on the display device and theGra
Displayed in a flick imageOf the wells of the microplate
Instructing means for instructing the position by an operation of an operator;
Based on the instruction from the instruction meansControl the capturing means
By capturing the minute object indicated by the
Move toBy controlling the sampling meansThe sampling
Transfer to take-off positionFine objectsCollectInstructed way
Control means for discharging the ink to the nozzle. [0008] [0009] [0010] [0011] [0012]BookAccording to the invention, the designated fine object is
By collecting in the wells of the indicated microplate.
This enables accurate and efficient separation and management of fine objects.
Wear. [0013] [0014] [0015] BRIEF DESCRIPTION OF THE DRAWINGS FIG.
It will be described with reference to FIG. FIG. 1 shows a fine structure according to an embodiment of the present invention.
FIG. 2 is a perspective view of a device for operating the object, and FIG.
FIG. 3 is a block diagram showing the configuration of the device for operating the fine object.
Lock diagram, FIG. 4 shows the processing function of the operating device for the fine object.
5 and 6 are functional block diagrams of the fine object.
FIG. 7 and FIG. 8 show the display screens of the fine objects.
Fig. 9 is a flow chart of the method of collection by ropes,
Flowchart of the sampling method without searching, FIGS. 10, 11,
FIG. 12 is a diagram showing a display screen of the operating device for the fine object.
You. First, operation of a fine object will be described with reference to FIGS.
The structure of the device will be described. In FIG. 1, the fine object
The operating device includes a main body 2 provided on a base 1 and a control unit.
And a personal computer 3. The body 2 is a cover
Each part described below is arranged in the case 2a.
ing. A base 5 is provided inside the cover case 2a.
Have been. Side frame 5a stands on the end of base 5
The upper frame is located above the side frame 5a.
The arm 5b is fixed. Side of side frame 5a
Is provided with a first XY table 6, and the first
The XY table 6 is provided with a translucent sample table 7.
Is being worn. The sample table 7 contains microorganisms, animal and plant cells.
Has a light-transmitting property containing a liquid sample containing fine objects such as
The sample container 8 is supported. The upper frame 5b has a transmission illumination 9 (first
An illumination unit) is provided.
The sample container 8 is irradiated with the illumination light reflected by. Sample volume
A camera 18 is provided at the bottom of the base 5 below the container 8.
I have. A fluorescent light 10 (second light) is provided on the side of the base 5.
Section) is provided, and the fluorescent light 10 is a half mirror 1
Illumination light is irradiated from below the sample container 8 via the reference numeral 5. Mosquito
The camera 18 is illuminated by the transmitted light 9 or the fluorescent light 10.
An image of the illuminated sample container 8 is taken. On the side of the base 5, an infrared laser light source 11 is provided.
(First laser device) and ultraviolet laser light source 12 (first
2 laser devices). Infrared laser light source
The infrared laser light emitted from 11 passes through the scanning optical system 13
Then, the light is irradiated upward by the mirror 17 to the sample container 8.
Light enters from below. By driving the scanning optical system 13,
The irradiation position of the infrared laser light is horizontal in the sample container 8.
Move in the direction. Therefore, the scanning optical system 13 uses the laser light
It is a means of transportation. Note that the scanning optical system 13 is used.
Instead, by driving the first XY table 6,
Infrared laser light is applied to the sample container 8 in the horizontal direction.
You may make it move oppositely. In this case,
One XY table 6 serves as a laser beam moving unit. The fine objects in the sample liquid in the sample container 8
By irradiating X-ray laser light, fine objects can be
It is trapped and captured by the light pressure of the user light. Scanning light
Drives the first XY table (or drives the first XY table)
By scanning with infrared laser light
The minute object to be captured in the sample container 8 with an infrared laser.
Irradiate the light and trap it, then contact it at any position
Can be moved. Infrared laser light source 11 and
The scanning optical system 13 scans a fine object with laser light.
This is so-called laser tweezers that
It is a capturing means for capturing. The ultraviolet light emitted from the ultraviolet laser light source 12
The line laser light is reflected by the mirror 16 and the sample container 8
From below. This ultraviolet laser light
Suitable for processing applications where small objects are locally destroyed.
By irradiating a fine object to be processed with X-ray laser light
Processing such as cutting of a fine object can be performed. UV light
The laser light source 12 is a processing means. Above the sample container 8, a sample supply pipette is provided.
The sample supply pipette 21 is XYZ
It can be moved by the table 22. Sample supply
The pipette 21 is connected to the dispensing mechanism 25,
The sample tank is connected to the mechanism 25 via pipes 26, 27 and 28.
29, the diluent tank 30 for washing and the waste tank 31
Has been continued. Using the suction / discharge mechanism of the dispensing mechanism 25
This allows the sample tank 29 and the diluent tank 30
The sample and diluent aspirated from the sample supply pipette 21
It is selectively discharged to the sample container 8 from the end. Sample volume after test
Extra sample remaining in the vessel 8 or dilution after being used for washing
The liquid is sucked by the dispensing mechanism 25 and stored in the waste liquid tank 31.
Collected. The XYZ table 2 is located near the sample container 8.
Micropipette for collection 23 movable by 4
But the suction port at the tip penetrates into the liquid in the sample container 8
It is arranged in force. Micropipette 23 is a dispensing mechanism
25 is connected to the suction / discharge function of the dispensing mechanism 25
Is used, the sample contained in the sample
A fine object located in the vicinity of the suction port of the
Inject / transfer and discharge from the tip of dispensing nozzle 36
Can be collected. Therefore, micropipette
G, dispensing mechanism 25 and dispensing nozzle 36 are fine objects
It is a collection means for collecting Below the dispensing mechanism 25, a second XY table is provided.
A bull 32 is provided. On the second XY table 32
Is a plate holder 33 which is a microplate supporting portion.
Is mounted on the upper surface of the plate holder 33.
Micro with well 35, a container for a number of fine objects
A plate 34 is placed. Second XY table 3
2 is driven and the microphone located immediately below the dispensing nozzle 36
By moving the plate 34 horizontally, an arbitrary c
A fine object discharged from the dispensing nozzle 36 is collected in the well 35.
Can be tolerated. Next, referring to FIG. 3, a device for operating a fine object
The configuration of the control system will be described. Camera 18, first illumination unit
9 and the second illumination unit 10 according to the instruction of the CPU 43.
To image a fine object in the sample container 8. The dispensing controller 41 has a dispensing mechanism 25
Control behavior. The scanning optical system controller 42 scans light
By controlling the science system 13, the infrared ray
The user light is moved in the sample container 8. CPU 43 controls
Means, storage means RAM44, ROM45,
Based on programs and data stored in the file device 46,
Then, various processes and calculations are executed. RAM 44 is CP
For temporarily storing data during U43 processing operation
Memory. ROM 45 is necessary for various processes and operations
Memorize various programs. The file device 46 stores programs and various
Data, for example, an image of a fine object captured by the camera 18
And the classification results of these fine objects. Memory
The medium drive device 47 stores necessary programs and data.
Writes / reads to / from an external storage medium. External memory
The medium 48 is a storage medium such as a floppy disk,
Used for data storage. The display monitor 49 displays
It is a device that displays screens during operation and input,
Displays an image of a fine object. The input unit 50 includes a keyboard and a mouse.
It is an inting device that can store operation commands and data.
In addition to input, fine objects displayed on the display monitor 49
The operator specifies the target of collection from within
Next, the microplate wafer in which this fine object is stored
Is instructed on the screen by the operator. sand
That is, the input unit 50 is an instruction unit. Instructor
As a step, besides this, touch panel, joystick
Etc. can be used. Next, referring to FIG. 4, a device for operating a fine object
Will be described. In FIG. 4, image processing
Means 51, capture operation control means 52, collection operation control means 5
3. The classification processing means 54 and the display processing means 55
3 is processing means (processing function). Ma
In addition, each storage unit described below stores RA as storage means.
M44, ROM 45, and file device 46
It shows the contents. The image processing means 51 is
From the image of the fine object in the sample container 8 taken,
A fine object is recognized by image processing. So the image
Processing means 51 searches for a target fine object in the sample
It is a search means. The reference image storage unit 56 stores the search condition.
The reference image used as the case is stored. Search condition memory
The unit 57 is a fine object in the field of view obtained by the image processing.
Performs search processing to identify the target minute object from the
The search conditions at the time of retrieval are stored. Sampling fine object image storage unit 5
8 stores an image of the searched and collected fine object
You. The microplate information storage unit 59 stores the collected fine
Array of wells in a microplate to accommodate small objects
Is stored. The collection result storage unit 60 stores the collected
The data on the type of the fine object
It is stored in association with the information of the well. Next, the contents of the above processing functions will be described.
You. The image data of the sample imaged by the camera 18 is
Image processing is performed by the image processing unit 51. And this
Image of each fine object in the field of view obtained by image processing
Matches the search conditions for identifying the target microscopic object.
The reference image storage unit 56 and the search condition storage unit
The data is compared with the data of No. 57 and determined. Used here
As a search condition, an image stored as a reference image
Based on agreement, based on dimensions, shape, color,
In addition, the movement speed and trajectory of fine objects in the field of view
Image information such as conditions set based on motion patterns
Various search conditions that can be set based on the search conditions are set. turtle
18 and the image processing means 51
Search means to search for the target micro object based on
ing. The target fine object searched by the above process
To the fine object identified as
By irradiating infrared laser light, fine objects
Trapped by Capture operation by this trap
Is controlled by the scanning optical system
Roller 42, first laser device 11, second laser device 1
2, performed by controlling the first XY table 6
It is. Reference of a fine object used in this search processing
The image is stored in a reference image storage unit 56 as storage means.
You. Dimensions, shapes, colors, gauges used as search conditions
Data such as traces and movement patterns such as moving speed are input
Input from the unit 50 and stored in the search condition storage unit 57
You. Also, based on the search results,
The fine object image data is stored in the collected fine object image storage unit 58.
It is memorized. The fine object searched by the image processing is
When collecting multiple kinds of fine objects,
Is subjected to classification processing assuming classification management after collection.
That is, the same sample is searched based on a plurality of search conditions.
Which micro-objects are being searched and collected
Which well of the plate corresponds to the search condition
A decision is made whether to accommodate the fine object. This decision
Use schedule stored in the microplate information storage unit 59
Classification processing means 5 based on the information of the microplate
4 done. Then, based on this decision, a search and a search are performed.
The microscopic pipette 2 removes the taken fine object from the sample container 8.
3 and collect the microscopic object
Process of dispensing to the plate 34, that is,
Store in designated well 35 of microplate 34
Dispensing process. These processes are performed by the collection operation control unit.
53, the second XY table 32, the XYZ table
Control the dispensing controller 42
Will be In the above processing, the image picked up by the camera 18 is
The displayed image is subjected to display processing by display processing means 55, and is displayed on a display monitor.
49 is displayed on the screen. On this screen, the input unit 50
By pointing the mouse at a specific fine object
Information that specifies the position of the specified minute object
It is sent to the operation control means 52. And based on this information
Operating optical system controller 42, first laser device 1
1, the second laser device 12 and the first XY table 6 are controlled.
Control the light pressure on the minute object indicated on the screen.
Can be trapped and captured. Also, a micro-computer displayed graphically
Use the mouse to locate a specific well on the rate screen
To specify the position of the indicated well.
A signal to be specified is sent to the sampling operation control means 53. This information
Based on the information, the second XY table 32, the XYZ table
By controlling the dispensing controller 42 and the dispensing controller 42
The specified well is positioned immediately below the dispensing nozzle 36.
And discharge a specific fine object into this well to collect
You. That is, from the input unit 50 as the instruction means
Capture means, laser light moving means,
And sampling means are controlled by the CPU 43 as control means.
The specified micro object to the specified web
It can be collected by discharging into the container. Next, referring to FIG. 5, a device for operating a fine object
Will be described. Figure 5 is the main menu
Is a display screen showing five types of operation buttons on the screen.
61 to 65 are displayed. Automatic operation ON, automatic operation OF
The operation buttons 61 and 62 of F perform an operation of collecting a fine object.
This is the operation button to be selected. Of these, automatic operation ON
When the operation button 61 is selected, when collecting a fine object,
Automatically search for fine objects in a sample based on set search conditions
The mode to search is selected. In addition, automatic search OFF operation
When the operation button 62 is selected, the automatic search function is stopped,
An operator recognizes a specific minute object on the screen of the display monitor 49.
Separately, the mode specified by the instruction means is selected. Select the operation button 63 for manual operation.
To search, capture, move, and collect minute objects
A mode in which all operations are performed manually is selected, and file management is performed.
And the operation buttons 64 and 65 for setting
Update of various data stored in each file device 46
File management mode and search conditions
Which setting mode is to be selected for each setting
You. Next, an automatic search will be described with reference to FIGS.
A mode for performing object sampling will be described. Screen of Figure 5
When the automatic search ON operation button 61 is selected above,
The screen shown in FIG. 6 is displayed on the display monitor. On the screen
Means that the image captured by the camera 18 is
The various fine objects 67 are displayed. Also display
The frame 68 has the object name, color, size, and characteristics of the movement pattern.
Along with the recognition result such as data,
Is displayed.
You. Further, a microplate graphic is displayed on the screen.
Image 69 is displayed.
It is possible to know the sample storage status of the plate. Ie already
The stored well 70 containing the sample is displayed in color.
The empty well 71 is displayed as blank. Stop
When the operation buttons 72 and 73 indicating interruption are selected,
The collection process can be stopped or interrupted. Next, sampling of a fine object by the automatic search mode is performed.
7 and 8 with reference to the drawings.
It will be explained along the line. In FIG. 2, first, the dispensing mechanism 25
The diluent is sucked from the diluent tank 30 by the
The sample on the sample table 7 discharged from the supply pipette 21
The container 8 is washed (ST1). Next, the dispensing mechanism 25
Therefore, a sample including a fine object is sucked from the sample tank 29.
And discharged from the sample supply pipette 21 to
(ST2). After that, the camera 18
The sample containing the liquid is imaged (ST3), and the obtained image is
Recognize and detect target minute objects by image processing from inside
A search process is performed (ST4). That is, from inside the sample
A target fine object is identified. Here, the search target
If you do not find any target fine objects in the sample
If so, the process returns to the first ST1 and the same processing is performed for a new sample.
Repeat the process. When a target fine object is found
(ST5), the found fine object, that is, the search target
Sampling of fine object image data of fine object image storage unit 58
(ST6), and the infrared laser beam is
By irradiating the identified fine object of interest
Trapped and captured by light pressure with laser tweezers,
Transfer to the sampling position of the micropipette 23 (ST
7). Then, the well containing the fine object
The position is determined by the classification processing means 54 (ST8).
So that the prepared well is located immediately below the dispensing nozzle 36.
By driving the second XY table 32, the micro plate 3
4 is positioned (ST9). Then XYZ table 2
4 to move the tip of the micropipette 23 to the sampling position
It moves (ST10) and drives the dispensing mechanism 25 to catch
The microscopic pipette 23
It is collected, discharged and stored in a predetermined well 35 (ST
11). At this time, the collected
The well position and the number of the image stored in ST6.
The result is stored in the result storage unit 60 (ST12). That is, collection
Information of the microscopic object associated with the containing well
I do. After this, it was determined whether the required number of fine objects had been collected.
It is determined (ST13), and if not collected, returns to ST3.
Repeat the following steps and make sure that
If the sample has been collected, clean the sample table 7 again.
(ST14), the collection process ends. Next, collection of fine objects without automatic search
The method will be described with reference to FIGS. First
Select automatic search OFF (see Fig. 5) in the main menu
By doing so, the screen shown in FIG. 10 is displayed (ST
21). This screen displays the same information as when the automatic search is ON.
In the image frame 66, an image of the minute object 67 and at the same time
Graphic image 69 showing black plate on the same screen
Will be displayed. At this time, the display frame 74
Request to specify the fine object to be collected and the well to contain
Is displayed, and a cursor 75 is displayed.
The state of the minute object to be collected and the state of waiting for the instruction of the well to contain
(ST22). The selection buttons 76, 77, 78
Indicates the cleaning of the sample container 8 and a new sample
Selection button to select the end of feed and collection operations
Tan. Thereafter, as shown in FIG.
Cursor to a specific fine object 67a from among the fine objects
Fines collected by clicking on
The pointing of the object is performed, and then the cursor 75 is moved to a specific place.
To the location of the container
Well instructions are made. Then confirm that there was an instruction.
(ST23), and subsequently instructed by image processing
A target fine object is recognized (ST24). This allows
The image of the recognized fine object and the recognition result are collected fine objects
It is stored in the image storage unit 58 (ST25). In addition, fine
For fine objects such as object name, color, size, feature data, etc.
The recognition result is, as shown in FIG.
Is displayed in the display frame 79 together with what kind of fine object is collected.
You can check if it is taken. Next, the target fine object is
And transfer it to the sampling position by the micropipette 23
(ST26). Next, the second XY table 32 is driven.
To position the microplate 34 (ST2).
7) Driving the XYZ table 24 to make a micropipette
23 is moved to the sampling position (ST28). this
Then, the desired fine object is sucked with the micropipette 23.
Discharge is performed to the designated predetermined well 35 (ST29).
Then, it is stored in the position of the accommodated well 35 and ST25.
The number of the captured image is stored in the collection result storage unit 60 (ST
30). Thereafter, returning to ST21, the same sampling step is performed.
Repeat steps. During the sampling process, the automatic sampling is displayed on the display frame 80.
A message will be displayed notifying you that you are busy
Next, the position of the receiving well is shown in the microplate graph.
Displayed on the display screen 69 by different colors, etc.
(See well number 2-1 indicated by an arrow in FIG. 12),
Information on the progress of sampling work, information on fine objects to be collected, and
The containment location can be checked visually at all times, and the
It is possible to perform a series of operations up to
Wear. According to the invention of this embodiment, fine particles in the liquid
Camera that captures small objects and images of captured fine objects
Display means for displaying, and a fine object in the liquid by a laser beam.
And a laser for moving a laser beam
Light moving means, and the displayed fine object is
And move it to locate the identified minute object.
It can be moved to a fixed position without contact. Preferably, the captured fine object is collected.
By providing a sampling means, a fine object can be combined with a liquid.
Can also be sampled and pointed at the indicated fine objects.
By collecting in the wells of the microplate
Separation management of fine objects can be performed accurately and efficiently
You. More preferably, the microplate
When indicating the position of the file,
The operability is improved by indicating the position of the black plate.
Image of a fine object captured by a camera
And graphic image of microplate at the same time
Make it easier to classify fine objects
Can be. In the present embodiment, the searched purpose
Laser tweezers when collecting microscopic objects
An example of capturing and moving is shown.
Without using the tweezers, the first XY table 6 or
Drives the XYZ table 24 to make the sample container 8 micro
By moving the pipette 23 relative to the pipette 23,
Target fine object directly to the tip of the micropipette 23
You may make it move and collect it. [0057] According to the present invention, fine objects in a liquid can be captured.
Image with the camera, display the image, and
By capturing and moving fine objects with optical tweezers
Can efficiently collect fine objects without contact
You.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of a fine object operating device according to one embodiment of the present invention; FIG. 2 is a side view of a fine object operating device according to one embodiment of the present invention; FIG. 4 is a block diagram showing a configuration of a fine object operating device according to one embodiment of the present invention. FIG. 4 is a functional block diagram showing processing functions of the fine object operating device according to one embodiment of the present invention. FIG. 6 is a diagram showing a display screen of a device for manipulating a fine object according to one embodiment of the present invention. FIG. 6 is a diagram showing a display screen of a device for manipulating a fine object according to one embodiment of the present invention. FIG. 8 is a flowchart of a sampling method by an automatic search for a fine object according to an embodiment of the present invention. FIG. 8 is a flowchart of a sampling method by an automatic search for a minute object according to an embodiment of the present invention. FIG. 10 is a flowchart of a sampling method that does not rely on automatic search for an object. FIG. 11 is a view showing a display screen of a fine object operation device. FIG. 11 is a view showing a display screen of a fine object operation device according to one embodiment of the present invention. FIG. 12 is an operation of a fine object according to one embodiment of the present invention. 6 shows a display screen of the apparatus [Description of References] 6 First XY table 7 Sample table 8 Sample container 11 Infrared laser light source (first laser device) 12 Ultraviolet laser light source (second laser device) 13 Scanning optical system 18 Camera 23 Micropipette 25 Dispensing mechanism 33 Plate holder 34 Microplate 35 Well 36 Dispensing nozzle 43 CPU 49 Display monitor (display device) 51 Image processing means

────────────────────────────────────────────────── ─── Continuation of front page (56) References JP-A-9-280375 (JP, A) JP-A-4-354532 (JP, A) JP-A-2000-202788 (JP, A) JP-A-2000-206011 ( JP, A) JP-A-2000-206057 (JP, A) JP-A-5-88107 (JP, A) JP-A-10-507518 (JP, A) JP-A-7-505710 (JP, A) Patent 2544520 (JP, A) JP, B2) WO 97/29355 (WO, A1) (58) Fields investigated (Int. Cl. ⁷ , DB name) G01N 1/00-1/44 B25J 7/00 G01N 35/00-35/10 B01J 19/00-19/32 C12M 1/00-1/42 G06T 1/00 G02B 21/32 JICST file (JOIS)

Claims (1)

(57) [Claims] [Claim 1] A camera for imaging a fine object in a liquid stored in a container, an image captured by the camera, and a microphone
A display device for simultaneously <br/> displayed on the same screen the graphic image of Ropureto, the microplate support portion for supporting the microplate in which a plurality of wells are formed, the
The container is irradiated with laser light and the light pressure of the laser light
To capture fine objects in liquid and transfer them to the sampling position
Means, and a sampling means for sucking the fine object transferred to the collection position and discharging it to the wells of the microplate,
The fine object on the image displayed on the display device and the
Instruction means for instructing the position of the well of the microplate displayed by the graphic image by an operation of an operator,
Controlling the capturing means based on an instruction from the instruction means
By capturing the fine object indicated by performing
It is moved to a position, before by controlling the sampling means
A device for operating a fine object, comprising a control means for collecting the fine object transferred to the recording position and discharging the fine object to a designated well.