JP2008152044A - Manipulator system, manipulator control program, and suction method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a manipulator system capable of surely, easily and rapidly replacing a minute sample, a manipulator control program and a suction method. <P>SOLUTION: The manipulator system includes a sample table 7, an automatic pipet 6, an XYZ stage 51, a microcamera 4 and a PC3 (personal computer). The PC 3 controls the automatic pipet 6 to suck a cell into the hollow part of a micro pipet 61 and discharge the sucked cell. Furthermore, the PC 3 controls the XYZ stage 51 to drive the micro pipet 61. Thus, the PC 3 controls the automatic pipet 6 and the XYZ stage 51 to alternately suck a cell culture solution in a Petri dish 8 and air, whereby a plurality of cells is consecutively sucked in such a state that they are separated one by one and discharged in such a state that they are separated one by one. Therefore, operation to mount the cell is rapidly and surely performed. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a manipulator system suitable for inhaling cells and the like, a manipulator control program, and an inhalation method.

Conventionally, in the field of medicine and the like, the operation of introducing cells one by one from a culture device such as a petri dish into a detector such as a chip is performed by using a device such as a pipette that manually sucks and discharges a cell solution. The operator was doing it manually while watching.
By the way, in recent years, as shown in Patent Document 1, a manipulator is used when a target sample is very minute and requires high-precision positioning, such as when a substance is injected into a cell.
JP 2005-118905 A

However, in order to introduce cells one by one into a detector or the like as described above, it is difficult to reliably inhale cells one by one, and skill is required for handling pipettes and the like. Further, if a manipulator as shown in Patent Document 1 is used, the operation of bringing the pipette closer to the cells is ensured, but the aspiration operation is still difficult, and further, for example, one cell at a time such as a 16-well plate. When it was necessary to introduce a large number, the above work had to be repeated, which required labor and time.
The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a manipulator system, a manipulator control program, and an inhalation method that can reliably, easily and quickly transfer a micro sample or the like.

  In order to solve the above-mentioned problem, a manipulator system according to claim 1 of the present invention has a pipe part having an open tip, suction means for sucking liquid into the hollow part from the tip, and a gas at the tip of the pipe part. Driving means for relatively moving in, and sucking the liquid by the suction means, forming a liquid layer in the hollow portion of the tube portion, and then relatively moving the tip of the tube portion into the gas by the driving means, Inhalation control means for forming an air layer on the distal end side of the hollow portion following the liquid layer by inhaling gas by the inhalation means.

The relative movement means that either the liquid side (a container or the like containing the liquid) or the tip of the pipe part may be moved.
A manipulator system according to a second aspect of the present invention is the manipulator system according to the first aspect, wherein the inhaling means is configured to discharge the inhaled material sucked into the hollow portion, and the driving means has the tip of the pipe portion at the inhaled material. The discharge means discharges the inhaled substance by the discharge means and the relative movement of the tip of the tube portion by the drive means alternately. Discharge control means to perform is provided.

  The manipulator system according to a third aspect of the present invention is the manipulator system according to the second aspect, wherein the inhaling means is configured to discharge the inhaled material sucked into the hollow portion, and the driving means has the tip of the pipe portion at the inhaled material. The control means is configured to be able to move relative to a container serving as a discharge destination of the liquid, and the control means discharges the inhaled substance by the discharge means and the relative movement of the tip of the tube portion by the drive means And are alternately performed.

A manipulator system according to a fourth aspect of the present invention is the manipulator system according to any one of the first to third aspects, wherein the liquid contains an insoluble substance, and the inhalation control means causes the inhalation means to inhale the insoluble substance, The liquid layer containing the insoluble substance is formed.
A manipulator system according to a fifth aspect of the present invention is the manipulator system according to the fourth aspect, wherein the drive means is configured to be able to relatively move the tip of the tube portion to a designated position in the liquid. The means is characterized in that the tip of the tube portion is moved relative to the vicinity of the insoluble substance by the driving means, and the insoluble substance is sucked by the suction means.

The manipulator system according to claim 6 of the present invention is the manipulator system according to claim 5, wherein the imaging means for imaging the liquid and the recognition means for recognizing insoluble substances in the liquid based on the image signal output from the imaging means. Position calculating means for calculating the position of the insoluble substance recognized by the recognition means, the suction control means based on the position information of the insoluble substance output from the position calculating means. The tip of the tube part is moved relative to the vicinity of the insoluble substance.
The manipulator system according to claim 7 of the present invention is characterized in that in any one of claims 4 to 6, the insoluble substance is a cell.

  According to an eighth aspect of the present invention, there is provided a manipulator control program having a pipe portion having an open end, suction means for sucking liquid into the hollow portion from the tip, and drive means for relatively moving the tip of the pipe portion into the gas. And a computer for controlling a manipulator comprising: a liquid suction step for sucking liquid by the suction means and forming a liquid layer in the hollow portion of the pipe portion; and the pipe portion by the driving means. A gas moving step for relatively moving the tip of the gas into the gas, and a gas suction step for forming a gas layer on the tip side of the hollow portion following the liquid layer by sucking the gas by the suction means. A program for causing the computer to execute processing is included.

According to a ninth aspect of the present invention, the manipulator control program according to the ninth aspect of the present invention includes, in the eighth aspect, after the gas suction step, a liquid movement step that relatively moves the tip of the tube portion into the liquid again. The liquid moving step is repeated.
An inhalation method according to claim 10 of the present invention is an inhalation method in which a liquid is sucked by an inhalation device having a pipe portion having an open tip and sucking liquid from the tip into a hollow portion, wherein the liquid and the gas are alternated. By inhaling, a plurality of liquid layers and gas layers are alternately formed in the hollow portion.

  According to the present invention, a plurality of cells or a plurality of types of liquids can be continuously inhaled in a separated state, and the inhalation operation can be speeded up. Similarly, the discharge operation can be performed quickly.

Next, embodiments of the present invention will be described with reference to the drawings.
FIG. 1 is a schematic view showing a manipulator system according to the present embodiment, FIG. 2 is a view showing an example of a channel chip 1 ((a) is a plan view, and (b) is a view for explaining a laminated structure. 3 is a diagram showing an outline of the automatic pipette 6, FIG. 4 is a diagram showing a configuration of each part of the automatic pipette 6, ((a) is a diagram showing the pump 63, (b). FIG. 5 is a flowchart showing processing by the PC 3 at the time of cell inhalation, FIG. 6 is a flowchart showing processing by the PC 3 at the time of cell discharge, and FIG. 7 is a cell culture. It is a schematic diagram which shows a mode that a solution layer and an air layer are formed alternately.

The manipulator system shown in FIG. 1 includes a sample table 7, an automatic pipette 6, an XYZ stage 51, a microscope camera 4, and a PC 3 (personal computer).
The sample table 7 is connected to an X stage 52 having a driving mechanism in the X direction (horizontal direction) (integrated with the movable part), and the table surface is movable in the X direction. In FIG. 1, a petri dish 8 containing a flow path chip 1 and a cell culture solution containing cells after cell culture is placed on the sample table 7. The X stage 52 that drives the sample table 7 moves the flow path chip 1 and the petri dish 8 under the field of view of the microscope camera 4 in response to a drive signal from the PC 3. The sample table 7 may use an XY stage.

  The channel chip 1 is a container that is a discharge destination of cells sucked from the petri dish 8 in the present embodiment, and specifically, the one shown in FIG. 2 is used. The flow path chip 1 in FIG. 2 is obtained by laminating a film electrode 14 (formed by laminating Pt and Ti) and a photosensitive resin 12 in a predetermined pattern on a substrate 11 by a lithography technique. By removing a part of the photosensitive resin 12, four recesses 2 are provided side by side, and each recess 2 forms a pair of solution tanks 21 and 22 and a fine channel 23 that communicates with this. . A pair of membrane electrodes 14, 14 are formed on the bottom surfaces of the solution vessels 21, 22, and are introduced into one of the solution vessels 21, 22 by applying a voltage between the membrane electrodes 14, 14. The solution thus obtained is driven toward the other solution tank through the fine channel 23. A narrow portion 24 having a narrower width than other portions is formed in the center of the fine channel 23 in the flow direction, and cells contained in the solution are sandwiched between the narrow portions 24 and crushed. In FIG. 1, reference numeral 9 denotes a connector. The end of the channel chip 1 where the membrane electrode 14 is exposed is attached to the connector 9 and is electrically connected to a power source (not shown).

Returning to the description of FIG. 1, the automatic pipette 6 receives an inhalation signal or an ejection signal from the PC 3 and inhales or ejects the cell solution, and corresponds to the inhalation unit and the ejection unit of the present invention. In the present embodiment, as shown in FIG. 3, a pump drive device 65, a coupling 64, a pump 63, a tube 62, and a micropipette 61 are provided.
The pump driving device 65 is composed of a stepping motor and a driver for driving the stepping motor, and the drive shaft of the stepping motor rotates based on the suction command or the discharge command input from the PC 3. The rotational driving force is transmitted to the pump 63 shown in FIG.

  In the pump 63 of FIG. 4, a connecting shaft 63 a coupled to the coupling 64 is rotatably supported inside the housing 63 b (not shown), and rotates according to the rotation of the pump driving device 65. The other end of the connecting shaft 63a is coaxially connected to the piston 63c via a screw mechanism inside the housing 63b. Then, the piston 63c is moved in the axial direction by the screw mechanism according to the rotation angle of the connection shaft 63a, whereby the hollow portion of the cylinder 63d is depressurized or increased in pressure. A tubular micropipette 61 is connected to the tip of the cylinder 63d via a tube 62, and the cell solution is sucked into the hollow part of the micropipette 61 by decreasing the internal pressure of the cylinder 63d, and is sucked into the hollow part by increasing the internal pressure of the cylinder 63d. The detailed solution is discharged. In FIG. 4, reference numeral 61a denotes a glass capillary.

The micropipette 61 constituting the automatic pipette corresponds to the tube portion of the present invention.
In the automatic pipette 6, the micropipette 61 is supported on the XYZ stage 51 via a pipette holder (not shown). The XYZ stage 51 includes an X-axis drive unit that drives in the X direction (horizontal direction), a Y-axis drive unit that drives in the Y direction (horizontal direction perpendicular to the X direction), and a Z-axis drive that drives in the Z direction (vertical direction). The micropipette 61 is driven in accordance with a drive signal that is configured from a part and designates a position from the PC 3. This XYZ stage 51 corresponds to the driving means of the present invention. The configuration of the XYZ stage 51 is not particularly limited in the present invention. For example, the driving method may be any of a pneumatic driving method, a hydraulic (water pressure) driving method, an electromagnetic force driving method, an electric motor driving method, and a piezoelectric driving method. Good. Further, separate driving methods may be used for fine movement and coarse movement, and these may be combined.

  The microscope camera 4 is composed of an optical microscope and a camera, converts an optical image magnified by the optical microscope into an electrical image signal by the camera, and outputs it to the PC 3, which corresponds to the photographing means of the present invention. . In FIG. 1, the microscope camera 4 is installed above the sample table 7 so that the petri dish 8 or the channel chip 1 enters the field of view, but the configuration (arrangement) of the microscope is not limited.

  The PC 3 includes a CPU (central processing unit), a RAM (random-access memory), a ROM (read-only memory), various drivers, and the like. The PC 3 performs image processing based on the image signal input from the microscope camera 4, and outputs drive signals to the automatic pipette 6, the XYZ stage 51, and the X stage 52 based on the result of the image processing. Thus, in this embodiment, the cells are sucked one by one from the cell culture solution in the petri dish 8 until the sucked cells are discharged one by one into the solution tanks 22 of the four recesses 2 of the flow channel chip 1. Perform a series of transfer operations.

Next, the processing by the PC 3 and the accompanying cell inhaling operation of the entire apparatus will be described using the flowchart shown in FIG. The suction operation is realized by a subroutine program that is called and executed by the main program.
First, it is assumed that the petri dish 8 containing the channel chip 1 and the cell culture solution is placed at a predetermined position on the sample table 7, and the petri dish 8 is under the field of view of the microscope camera 4. It is assumed that the user has already set the number of cells to be inhaled. In this state, first, in step S101, an image signal is acquired from the microscope camera 4. In subsequent step S102, the image signal acquired from the microscope camera 4 in step S101 is subjected to image processing, a cell is recognized, and any one of the recognized plurality of cells is selected. Then, the coordinates of the selected cell are calculated, and the coordinates of the inhalation position are calculated based on this. The processing for recognizing the cells corresponds to the recognition means of the present invention, and the processing for calculating the cell coordinates corresponds to the position calculation means of the present invention. In the subsequent step S103, a command value to the XYZ stage 51 is calculated based on the coordinates calculated in step S102. Based on the command value calculated in step S103, first, a drive command in the X and Y directions is output to the XYZ stage 51 in step S104, and a drive end signal is input from the XYZ stage 51. In S105, a drive command in the Z direction is output so that the tip of the micropipette 61 can inhale the cell culture solution. Steps S104 and S105 correspond to the liquid movement step of the present invention.

  Next, when a drive end signal is input from the XYZ stage 51 in step S106, a cell inhalation command is output to the pump drive device 65. Thereby, the internal pressure of the cylinder 63d of the automatic pipette 6 is reduced, and the designated cells are sucked into the hollow portion of the micropipette 61. The inhalation command is a command value such that the inhalation amount of the cell culture solution containing cells is an amount greater than or equal to the volume of the cell. For example, predetermined inhalation amounts for various cell culture solutions according to the viscosity, specific gravity, etc. of the cells And Further, when the inhalation of the cell culture solution is repeated as described later, this may be changed for each inhalation. After the inhalation, the decompression is stopped, and the internal pressure of the cylinder 63d is maintained so that the position and shape of the cell culture solution layer containing the inhaled cells formed in the micropipette 61 are maintained. This step S106 corresponds to the liquid suction step of the present invention.

  In step S107, when a signal indicating completion of inhalation is input from the pump drive device 65, the drive command to the air inhalation position, that is, the tip of the micropipette 61 is separated upward from the liquid level of the cell culture solution. A drive command is output to the XYZ stage 51. As a result, the Z-axis drive unit of the XYZ stage 51 is operated to drive the micropipette 61 upward, so that the tip of the micropipette 61 is separated from the liquid surface. The upward driving amount is set to a predetermined value, for example. This step S107 corresponds to the gas moving step of the present invention.

  Next, when a driving end signal is input from the XYZ stage 51 in step S108, an air suction command is output to the pump driving device 65. As a result, the internal pressure of the cylinder 63d of the automatic pipette 6 is reduced, air is sucked into the micropipette 61, and an air layer is formed on the tip side of the cell culture solution layer. When the cell culture solution layer is formed before and after the air layer by inhaling a plurality of cells, the air inhalation amount is an inhalation amount that can clearly separate the cell culture solution layer. Use the defined value. This step S108 corresponds to the air suction step of the present invention.

  Thereafter, in step S109, it is determined whether or not the set number of inhalations have been inhaled. If the inhaled number has already been set (step S109: Yes), the inhalation process is terminated and the process returns to the main program. If the set number of inhalations has not yet been reached (step S109: No), the process returns to step S101 again, and the processes shown in steps S101 to S109 are repeated until the set number is reached. Thereby, as shown in the schematic diagram of FIG. 7, the cell culture solution layer and the air layer are alternately formed in the hollow portion of the micropipette by the set number (four in the figure). Become. The cell culture solution layer containing the cells corresponds to the liquid layer of the present invention, and the air layer corresponds to the gas layer of the present invention.

  The above processing by the PC 3 corresponds to the inhalation control means of the present invention. As described above, in order to inhale a plurality of cells, if a cell culture solution containing inhaled cells and air are alternately inhaled, and a plurality of cell culture solution layers and air layers are alternately formed, one by one Rather than inhaling cells and mounting them one by one on the channel chip 1 and repeating this for the target number, the inhalation operation can be performed continuously, which is quicker and more efficient. In addition, it is conceivable to inhale a plurality at a time without forming an air layer in the micropipette 61. In this case, however, cells stick together in the micropipette 61, and each one is successfully discharged at the time of discharge. Individual cells cannot be separated. In this regard, if an air layer is formed, each cell culture solution layer can be reliably separated in the micropipette 61 by the air layer, so that each cell can be discharged in a separated state. Further, if the layer to be separated is an air layer, it is not necessary to prepare a material for the separation layer in particular, and it can be easily formed.

Next, the processing by the PC 3 and the cell discharge operation of the entire apparatus accompanying this will be described using the flowchart shown in FIG. The discharge operation is realized by a subroutine program that is called and executed by the main program.
In a state where a plurality of cells are inhaled into the micropipette 61 and the automatic pipette 6 is lifted by a predetermined distance away from the liquid surface as described above, first, in step S201, the flow channel chip 1 is placed under the field of view of the microscope camera 4. The drive command for the sample table 7 is output to the X stage 52 so that the In the present embodiment, driving is performed by a predetermined amount.

  In the subsequent step S202, when the driving end signal of the sample table 7 is input, the image signal of the channel chip 1 is acquired from the microscope camera 4. Then, in subsequent step S203, image processing is performed to recognize a cell discharge position (mounting position) and calculate its coordinates. In the present embodiment, since the cells are mounted one by one in the four solution tanks 22 of the flow path chip 1 described above, the coordinates of each solution tank 22 are calculated and stored. At this time, the shape of the solution tank 22 or the pattern of the film electrode 14 is stored in advance, the pattern is extracted from the image signal, the solution tank or the electrode is recognized, and its coordinates are calculated. In subsequent step S204, command values to the XYZ stage 51 are calculated based on the coordinates of the four positions for mounting the cells calculated in step S203.

  Then, based on the command value calculated in step S204, first, in step S205, a drive command in the XY direction is output to the XYZ stage 51. After a drive end signal is input from the XYZ stage 51, in the subsequent step S206, the Z direction To the state where the cells can be discharged from the tip of the micropipette 61.

  When a drive end signal is input from the XYZ stage 51, the PC 3 outputs a discharge command for one cell to the automatic pipette 6 in the subsequent step S207. At this time, a discharge command for discharging a predetermined amount is output, but an image signal is acquired from the microscope camera 4 (step S208), and the image signal is subjected to image processing, whereby cells are discharged to a position to be mounted. It is determined whether or not. When it is determined that the cells have been ejected (step S208: Yes), the process proceeds to step S208. On the other hand, when it is determined that the cells are not discharged (step S208: No), the process returns to step S206, and the internal pressure of the cylinder 63d of the automatic pipette 6 is gradually increased until the discharge of the cells is confirmed.

In step S209, it is determined whether or not the number of ejections has become the number of inhalations, that is, whether or not all the inhaled cells have been ejected. If all of the inhaled cells have been ejected (step S209: Yes), the ejection operation is terminated and the main program Return to. On the other hand, if all the inhaled cells have not yet been discharged (step S209: No), steps S205 to S209 are repeated until all the cells have been discharged.
The above processing by the PC 3 corresponds to the discharge control means of the present invention. Thus, by continuously inhaling the cell culture solution layer with the air layer sandwiched in the micropipette 61, the cells can be continuously discharged in a state of being separated one by one. The discharge operation can be performed reliably.

Although the present invention has been described above, in the flow of FIG. 5, it may be determined from an image photographed by the microscope camera 4 whether or not cells are inhaled in step S106. In step S102, the cells to be inhaled are selected on the PC3 side after the cell recognition. For example, the recognition result is displayed to the user on the monitor, and the user wants to select the cells to be inhaled. The inhalation position may be calculated.
Furthermore, control such as driving of the sample table may be driven in a sequence.

In addition, the application of the present invention is not limited to the above-described embodiment. For example, the present invention is applied not only when inhaling a cell culture solution but also when inhaling a different type of liquid and transferring it to another container. The invention can be applied.
Further, as shown in FIG. 1, not only when the micropipette 61 is moved, the sample table 7 on which the petri dish 8 is placed is configured to move the sample table 7 in the X, Y and Z directions, A configuration may be adopted in which the micropipette 61 is moved in the Z direction by moving in the X and Y directions. When cells are inhaled from a container or the like having a certain depth, a camera may be further provided to confirm the position in the Z direction.

It is the schematic which shows the manipulator system of this embodiment. (A) is a top view of a flow-path chip | tip, (b) is a figure for demonstrating the laminated structure of a flow-path chip | tip. It is a figure which shows the outline of an automatic pipette. (A) is a figure which shows a pump, (b) is a figure which shows a micropipette. It is a flowchart which shows the process by PC at the time of cell inhalation. It is a flowchart which shows the process by PC at the time of cell discharge. It is a schematic diagram which shows a mode that a cell culture solution layer and an air layer are formed alternately.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Channel tip 2 Recessed part 4 Microscope camera 6 Automatic pipette 7 Sample table 8 Petri dish 9 Connector 11 Board | substrate 12 Photosensitive resin 14 Film-like electrode 21, 22 Solution tank 23 Fine channel 24 Narrow part 61 Micropipette 62 Tube 63 Pump 63a Connection shaft 63b Housing 63c Piston 63d Cylinder 64 Coupling 65 Pump drive device

Claims (10)

An inhalation means having a tube portion with an open tip, and sucking liquid from the tip into the hollow portion;
Drive means for relatively moving the tip of the tube portion into the gas;
The liquid is sucked by the suction means, a liquid layer is formed in the hollow portion of the pipe portion, the tip of the pipe portion is relatively moved into the gas by the driving means, and the gas is sucked by the suction means. Inhalation control means for forming an air layer on the distal end side of the hollow portion following the liquid layer,
A manipulator system comprising: The driving means is configured to be able to relatively move the tip of the tube portion into the liquid,
2. The suction control unit according to claim 1, wherein a plurality of liquid layers and gas layers are alternately formed in the hollow portion of the tube portion by alternately sucking the liquid and gas by the suction unit. Manipulator system characterized by The inhalation means is configured to be able to discharge the inhaled material inhaled into the hollow portion,
The drive means is configured to be able to move the tip of the tube portion relative to a container that is a discharge destination of the inhaled matter,
The manipulator system according to claim 2, further comprising: a discharge control unit that alternately discharges the inhaled material by the discharge unit and the relative movement of the distal end of the pipe portion by the driving unit. The liquid contains an insoluble substance,
The manipulator system according to any one of claims 1 to 3, wherein the inhalation control means causes the inhalation means to inhale the insoluble substance and forms the liquid layer containing the insoluble substance. The driving means is configured to be able to relatively move the tip of the tube portion to a designated position in the liquid,
5. The manipulator system according to claim 4, wherein the inhalation control unit causes the distal end of the pipe portion to move relative to the vicinity of the insoluble substance by the driving unit and inhales the insoluble substance by the inhalation unit. Photographing means for photographing the liquid;
Recognition means for recognizing an insoluble substance in the liquid based on an image signal output from the photographing means;
Position calculating means for calculating the position of the insoluble substance recognized by the recognition means,
The inhalation control means moves the tip of the tube portion relative to the vicinity of the insoluble substance by the driving means based on the position information of the insoluble substance output from the position calculating means. 5. The manipulator system according to 5.   The manipulator system according to claim 4, wherein the insoluble substance is a cell. Controlling a manipulator having a pipe portion having an open tip, suction means for sucking liquid from the tip into a hollow portion, drive means for relatively moving the tip of the pipe portion into gas, and a computer A program,
A liquid suction step for sucking liquid by the suction means and forming a liquid layer in the hollow portion of the tube portion; a gas movement step for relatively moving the tip of the tube portion into the gas by the driving means; and the suction means A program for causing the computer to execute a process comprising: a gas suction step of forming a gas layer on the distal end side of the hollow portion following the liquid layer by sucking the gas by Manipulator control program. After the gas suction step, the liquid moving step of relatively moving the tip of the tube portion into the liquid again,
The manipulator control program according to claim 8, wherein the steps from the liquid suction step to the liquid movement step are repeated. An inhalation method for inhaling a liquid by an inhalation device for inhaling a liquid from the distal end to a hollow portion having a tube portion with an open tip,
An inhalation method, wherein a plurality of liquid layers and gas layers are alternately formed in the hollow portion by alternately inhaling liquid and gas.

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