JP2010261929A - Microinjection apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microinjection apparatus, capable of improving processing efficiency, capable of simultaneously introducing a substance to be introduced to a plurality of cells. <P>SOLUTION: The microinjection apparatus introduces the substance to be introduced into a body to be introduced by injection needles 11. The microinjection apparatus includes a vessel position adjustment means 1, an imaging means 2, a position determination means 3, and a plurality of transport means 4. The vessel position adjustment means 1 adjusts the position of a vessel 12 stored in the body to be introduced. The imaging means 2 captures an internal image of the vessel 12, of which the position has been adjusted. The position determination means 3 determines the position of the body to be introduced from the image by the imaging means 2. The plurality of transport means 4 individually makes the plurality of injection needles 11 move, according to the positional information by the position determination means 3 for the respective injection needles 11. The transport means 4 has a moving mechanism, of at least not less than two degrees of freedom and uses a laminated-type piezoelectric element as a drive source of a moving mechanism having at least one or more degrees of freedom. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a microinjection apparatus for introducing an introduction substance such as a gene regulatory factor into a body to be introduced such as a cell with a minute injection needle.

  Typical methods for introducing a desired substance such as a gene regulatory factor into a cell include an electroporation method that is an electrical method, a lipofection method that is a chemical method, a vector method that is a biological method, and a machine. There are a microinjection method that is a typical method, a laser injection method that is an optical method, and the like.

  Among these, the electroporation method, which is an electrical method, damages the cell because the cell membrane is broken by a large current. In the case of the lipofection method, which is a chemical method, there are limitations on gene regulatory factors that can be introduced as introduction substances, and the introduction efficiency is low. In the case of the vector method, which is a biological method, there are limitations on the gene regulatory factors that can be introduced, and there are problems with safety. On the other hand, the microinjection method, which is a mechanical method, has a feature that an introduced substance can be reliably introduced into cells by controlling the injection position with high accuracy. As an example of this microinjection method, a method of introducing an introduced substance into cells using a capillary (a minute injection needle) has been proposed (Patent Document 1).

  As a technique for increasing the processing efficiency of the microinjection method, a microcapillary array in which a large number of injection needles are regularly arranged and a plurality of microchambers for fixing cells at positions corresponding to the injection needles in this array are arranged. There has also been proposed a method of injecting gene regulatory factors into a plurality of cells in a lump while facing a microchamber array (Patent Document 2).

  FIG. 19 shows a conventional example of an apparatus to which the microinjection method is applied. In this microinjection apparatus, a petri dish 90 containing cells to be introduced is gripped by a container mounting table 91, a local planar image inside the petri dish 90 is captured by an imaging unit 92, and the planar image is image processing unit 93. The position information of the cells is obtained by processing in step (1). By moving the container mounting table 91 by the horizontal biaxial direction container position adjusting means 94 made of an XY stage device, the cells are positioned so that the cells are positioned in the insertion direction of the injection needle 95. Next, the manipulator holding the injection needle 95 is moved in the insertion direction of the injection needle 95 by the injection needle transport means 97 having the Z stage device 96. In this way, the injection needle 95 is pierced into the positioned cell, and the introduction substance filled in the injection needle 95 is introduced into the cell. A series of these operations are automatically performed under the control of the control device 98. An ultrasonic motor or a ball screw is used for the injection needle conveying means 97.

Japanese Patent No. 2624719 Japanese Patent No. 3035608

  Examples of the method for introducing a gene regulatory factor such as DNA or protein into a cell include the above-described methods. However, a technique that achieves both reliability and efficiency has not yet been established. Among these methods, the microinjection method in which a gene regulatory factor is introduced into each cell is the most reliable method, but the operation requires skill and time, so that the processing efficiency is low. There is.

  In the method of collectively controlling the position of the injection needle for a plurality of cells as in the microcapillary array system disclosed in Patent Document 2, the shape, size, and elasticity of each cell are different. As a result, the injection needle is not pierced or many cells are destroyed. In addition, the microcapillary array method requires cells to be arranged at the same position as each injection needle of the microcapillary array, so it can only be applied to floating cells that can move in the culture solution, and adheres to the bottom of the medium or petri dish. It cannot be applied to cells that have been damaged.

  Therefore, in the apparatus to which the microinjection method is applied, improvement of the injection speed for injecting the introduction substance into the cell and improvement of the positioning speed of the manipulator have been attempted so far in order to improve the processing efficiency. Some of the current microinjection apparatuses realize the injection process of one cell in one second or less.

However, medical use requirements require that the type of substance introduced into the cell is not limited, that introduction efficiency is high, and that a large amount of substance-introduced cells can be supplied, and that particularly important substance-introduced cells can be supplied in large quantities. The request cannot be met at present.
Further, even if it is desired to use a plurality of injection needles, there is a problem in that a plurality of manipulators cannot be arranged because the size of a manipulator equipped with injection needles is large at present.

  For example, in the current microinjection apparatus as described above with reference to FIG. 19, for example, a manipulator equipped with an injection needle 95 is moved by moving the container position adjusting means 94 that supports the petri dish 90 containing the cells in order to position the cells. Since the cells are moved only in the direction in which the injection needle 95 is inserted, it is impossible to position a plurality of cells and simultaneously insert the injection needles into the cells.

  An object of the present invention is to provide a microinjection apparatus that can efficiently and surely introduce a substance to be introduced into an introduced body such as a cell with a plurality of injection needles and can improve the processing efficiency. is there.

  The microinjection device of the present invention is a microinjection device for introducing an introduced substance into a body to be introduced by inserting a minute injection needle filled with the introduced substance into the body to be introduced. A plurality of container position adjusting means for adjusting the position of the container accommodated therein, and a plurality of injection needles for individually moving the plurality of injection needles with respect to the introduction body inside the container whose position is adjusted by the container position adjusting means. A transfer means, an image pickup means for picking up an image inside the container whose position is adjusted by the container position adjustment means through an enlargement lens, and a position of the introducer is determined from the image obtained by the image pickup means. A position determination unit; and a control unit that causes each of the transfer units to move the injection needle in accordance with position information obtained by the position determination unit. The stage has at least two degrees of freedom, and has a movement mechanism for each degree of freedom, and among the movement mechanisms for each degree of freedom, a drive source of the movement mechanism for at least one degree of freedom includes a plurality of piezoelectric elements. A piezoelectric element laminate that is laminated and expands and contracts in the lamination direction is used.

According to this configuration, the position of the container containing the introducer is adjusted by the container position adjusting means, the planar view image inside the position-adjusted container is picked up by the imaging means, and the cell position is determined from the image by the position determining means. To determine the introduction body such as the cells in charge of each injection needle. At this time, the position of the introduced body can be recognized with high accuracy by locally enlarging the container with the magnifying lens and performing image processing on the position of the introduced body such as cells. In addition, for example, from the positional relationship in the image of the introducer such as the recognized cells, the introducer in charge of each transfer device can be determined. After determining the introduction body to be in charge of each injection needle in this way, each injection needle is moved by a plurality of individual conveying means, and the injection needle is inserted into each introduction body, so that each introduction body is inserted into each introduction body. Introduce introduced material. For this reason, the introduction substance can be simultaneously introduced into a plurality of introduction bodies such as cells, and the efficiency of the injection process can be improved. In addition, by filling at least two different kinds of different injection substances for each injection needle and sequentially introducing them into the same substance to be introduced, it is possible to introduce multiple kinds of introduction substances into the substance to be introduced at the same time. . Therefore, a plurality of kinds of introduction substances can be introduced without replacing the injection substance in the injection needle, and the efficiency of the injection process can be improved.
In particular, the conveying means has a moving mechanism having at least two degrees of freedom, and uses a stacked piezoelectric element as a drive source for the moving mechanism having at least one degree of freedom among the moving mechanisms having each degree of freedom. The size can be reduced, and a large number of conveying means can be arranged in a limited space around the container, so that a large number of injection needles can be used, and the efficiency of the injection process can be improved.

In the present invention, the moving mechanism using the piezoelectric element laminate in the transporting unit as a drive source arranges the plurality of piezoelectric element laminates in parallel, and the plurality of piezoelectric element laminates are stretched in the extending direction via the fastening member. May be connected in series.
Thus, when a plurality of piezoelectric element laminates are arranged in parallel and connected in series, a larger displacement can be obtained with a compact configuration. In order to enable the tip of the injection needle to move within the entire field of view in the image obtained by the imaging means, the movement distance of the injection needle by the conveying means is preferably 1 mm or more. By adopting the arrangement and connection configuration of the body, it is possible to secure a sufficient amount of movement necessary for the injection needle without requiring a large space.

  In the present invention, the moving mechanism using the piezoelectric element laminate in the transport unit as a drive source may include an expansion mechanism that expands and contracts the piezoelectric element laminate to a displacement in a direction perpendicular to the expansion and contraction direction. In the case of this configuration, it is more effective in securing the amount of movement necessary for the injection needle.

The enlargement mechanism may be a link mechanism. This link mechanism is composed of, for example, a plurality of links and a joint that constitutes a rotatable node that connects these links. Each joint is a fixed joint with a fixed position or a movable joint with a movable position.
With the link mechanism, the direction of displacement can be changed with a simple configuration, and the displacement can be greatly enlarged. Each joint may be a joint that rotatably couples components connected to each other via a rolling bearing or a sliding bearing. When a rolling bearing is used, it is possible to reduce the frictional resistance and to suppress the backlash by applying an appropriate preload to the rolling bearing, thereby realizing precise positioning. In addition, since there is no elastic deformation part, the design of the enlargement mechanism is easy.

The link mechanism serving as the enlarging mechanism has three links, one fixed joint that is fixed in position, and three movable joints that are movable in position, and converts the displacement in the expansion / contraction direction of the piezoelectric element laminate into an orthogonal direction. In addition, the displacement may be enlarged.
In this configuration, each of the joints is a joint that forms a rotatable node, and the center of rotation of the joint is relative to either the expansion / contraction direction of the piezoelectric element laminate or the direction orthogonal to the expansion / contraction direction. Even vertical. The other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via a second movable joint, A third link is connected to the tip of the second link via a third movable joint, and the tip of the third link is a movable part that is constrained to be movable only in a direction orthogonal to the expansion / contraction direction. The fixed joint is fixed in position with respect to the fixed end of the piezoelectric element laminate, and the first movable joint is movable integrally with the extensible end of the piezoelectric element laminate. The movable part at the tip of the third link becomes the displacement expansion output part of the link mechanism.
In the case of this configuration, it is not necessary to connect the three links at the same position, it is not necessary to arrange two joints side by side in the axial direction, and the thickness dimension of the link mechanism can be reduced.

The link mechanism serving as the enlarging mechanism has two links, one fixed joint that is fixed in position, and two movable joints that are movable in position, and converts the displacement in the expansion / contraction direction of the piezoelectric element laminate into an orthogonal direction. In addition, the displacement may be enlarged.
In this configuration, each of the joints is a joint that forms a rotatable node, and the center of rotation of the joint is relative to either the expansion / contraction direction of the piezoelectric element laminate or the direction orthogonal to the expansion / contraction direction. Even vertical. The other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via the second movable joint, and the fixed The joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is movable integrally with the extensible side end of the piezoelectric element laminate. The tip of the second link becomes the displacement expansion output unit of the link mechanism.
Also in this configuration, it is not necessary to connect the three links at the same position, it is not necessary to arrange two joints side by side in the axial direction, and the thickness dimension of the link mechanism can be reduced.

  In the present invention, the moving mechanism using the piezoelectric element laminate in the conveying means as a drive source includes a first expansion mechanism that expands the expansion and contraction of the piezoelectric element to a displacement perpendicular to the expansion and contraction direction, and the first expansion mechanism. It is good also as what has the 2nd expansion mechanism which expands the displacement expanded by the mechanism to the displacement of the expansion-contraction direction of a piezoelectric element. In the case of this configuration, it becomes more effective in securing the amount of movement necessary for the injection needle.

The first and second enlargement mechanisms may be link mechanisms. The displacement in the expansion / contraction direction of the piezoelectric element laminate is converted into an orthogonal direction by a link mechanism serving as a first expansion mechanism, the displacement is expanded, and the expanded displacement in the orthogonal direction is further converted into a second expansion mechanism. The link mechanism is converted into a displacement in the expansion / contraction direction of the piezoelectric element laminate, and the displacement is enlarged. Each link mechanism is composed of, for example, a plurality of links and joints that constitute rotatable nodes that connect the links. Each joint is a fixed joint with a fixed position or a movable joint with a movable position. It is assumed that the two link mechanisms constituting the first and second enlargement mechanisms are combined in two stages via a movable joint so that displacement is sequentially transmitted, for example.
Also in this configuration, each joint may be a joint that rotatably couples components connected to each other via a rolling bearing or a sliding bearing. When a rolling bearing is used, it is possible to reduce the frictional resistance and to suppress the backlash by applying an appropriate preload to the rolling bearing, thereby realizing precise positioning. Moreover, since there is no elastic deformation part, the design of the 1st and 2nd expansion mechanism is easy.

Each of the link mechanisms constituting the first and second expansion mechanisms includes three links, one fixed joint that is fixed in position, and three movable joints that are movable in position. The link mechanisms constituting the second expansion mechanism may be combined in two stages so that the displacement is sequentially transmitted via the movable joint.
Each of the joints is a joint that constitutes a rotatable node, and the center of rotation is perpendicular to both the expansion / contraction direction of the piezoelectric element laminate and the direction orthogonal to the expansion / contraction direction. is there. The other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via a second movable joint, The third link is connected to the tip of the second link via a third movable joint. The tip of the third link is a movable part that is constrained to move only in a direction orthogonal to the expansion and contraction direction.
In the link mechanism constituting the first expansion mechanism, the fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided on the expansion / contraction side of the piezoelectric element laminate. It can move together with the end.
The link mechanism constituting the second enlargement mechanism is provided in a posture in which the direction of 90 ° is changed around the central axis parallel to the rotation center of each joint with respect to the link mechanism constituting the first enlargement mechanism, The fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided in the movable portion in the link mechanism constituting the first expansion mechanism. The movable portion of the third movable joint of the link mechanism constituting the second expansion mechanism serves as a displacement expansion output portion in the expansion / contraction direction of the piezoelectric element laminate.

Each of the link mechanisms constituting the first and second enlargement mechanisms includes two links, a fixed position-fixing joint, and two position-movable joints. The link mechanisms constituting the second expansion mechanism may be combined in two stages so that the displacement is sequentially transmitted via the movable joint.
Each of the joints is a joint that constitutes a rotatable node, and the center of rotation is perpendicular to both the expansion / contraction direction of the piezoelectric element laminate and the direction orthogonal to the expansion / contraction direction. is there. Each link mechanism is configured such that the other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via the second movable joint. Link.
In the link mechanism constituting the first expansion mechanism, the fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided on the expansion / contraction side of the piezoelectric element laminate. It can move together with the end.
The link mechanism constituting the second enlargement mechanism is provided in a posture in which the direction of 90 ° is changed around the central axis parallel to the rotation center of each joint with respect to the link mechanism constituting the first enlargement mechanism, The fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided at the tip of the second link in the link mechanism constituting the first expansion mechanism. The tip of the second link of the link mechanism that constitutes the second expansion mechanism serves as a displacement expansion output section in the expansion / contraction direction of the piezoelectric element laminate.

The link mechanism includes a crank / slider mechanism including one fixed joint, two movable joints, and two links. The crank / slider mechanism is configured to change the displacement of the piezoelectric element in the extension direction. It is also possible to convert the displacement to an arbitrary direction on the circumference of the fixed joint through the joint and the two links and further increase the displacement.
In this configuration, for example, the fixed joint is arranged on the same line in the moving direction of the first movable joint. In this configuration, each of the joints is a joint that forms a rotatable node, and the center of rotation of the joint is relative to either the expansion / contraction direction of the piezoelectric element laminate or the direction orthogonal to the expansion / contraction direction. Even vertical. By setting the output unit at an arbitrary position on the circumference of the fixed joint, the displacement can be enlarged in an arbitrary direction. Therefore, the number of parts can be reduced and the size can be reduced.

  The crank / slider mechanism has a configuration in which a fixed joint is arranged on the same line in the moving direction of the movable joint, but may be an offset crank mechanism in which the fixed joint is not on the same line in the moving direction of the movable joint. In this case, the rotation angle of the fixed joint differs depending on the reciprocation with respect to the reciprocation of the first movable joint. Therefore, by selecting a point where the rotation angle of the fixed joint is large with respect to the movement amount of the first movable joint, the displacement enlargement ratio can be increased without increasing the size of the link.

  In this invention, the moving mechanism that moves the needle support member that supports the injection needle in the insertion direction of the injection needle is a movement mechanism that uses the piezoelectric element laminate as a drive source, and a part of the movement mechanism It is good also as what has a vibration drive means to give the vibration of an insertion direction to the said needle support member by carrying out the vibration drive of this piezoelectric element. In the case of this configuration, the injection needle can be smoothly pierced into the derivative and the injection needle can be positioned at high speed.

  The vibration driving means may superimpose a positioning signal for positioning the needle support member and a vibration drive signal for vibrating the needle support member in the insertion direction and apply the superimposed signal to the partial piezoelectric elements. In the case of this configuration, a part of the piezoelectric element laminated body can be shared for vibration application and positioning.

  The piezoelectric element to be vibrated by the vibration driving means may be a piezoelectric element located at a connection side end with respect to the needle support member in the piezoelectric element laminate. In the case of this configuration, displacement due to vibration generated in the piezoelectric element can be efficiently transmitted to the injection needle.

  The vibration drive means may include frequency variable means for switching the frequency of vibration. In the case of this configuration, the frequency of the vibration can be switched according to the type of the body to be introduced into which the injection needle is inserted, and the insertion needle can be smoothly inserted accordingly.

  In the case where the vibration driving means is provided, a connection body in which one piezoelectric element laminated body, which is a part of the piezoelectric elements driven by vibration, and another piezoelectric element laminated body are linearly connected is connected to another piezoelectric element. You may arrange | position in parallel with an element laminated body, and may connect the connection body arrange | positioned in parallel with each other, and another piezoelectric element laminated body in series in the expansion-contraction direction via a fastening member. Thereby, it is possible to secure a sufficient amount of movement necessary for the injection needle with a compact configuration, and it is also possible to impart vibration.

  In the present invention, the introduced substance may be a cell, and the introduced substance may be a gene regulatory factor. The gene regulatory factor is, for example, DNA or protein. In the case of injecting a gene regulatory factor into such a cell, the advantage of being able to efficiently and surely perform with a plurality of injection needles in this invention and improving the processing efficiency is effectively exhibited. .

  The microinjection device of the present invention is a microinjection device for introducing an introduced substance into a body to be introduced by inserting a minute injection needle filled with the introduced substance into the body to be introduced. A plurality of container position adjusting means for adjusting the position of the container accommodated therein, and a plurality of injection needles for individually moving the plurality of injection needles with respect to the introduction body inside the container whose position is adjusted by the container position adjusting means. A transfer means, an image pickup means for picking up an image inside the container whose position is adjusted by the container position adjustment means through an enlargement lens, and a position of the introducer is determined from the image obtained by the image pickup means. A position determination unit; and a control unit that causes each of the transfer units to move the injection needle in accordance with position information obtained by the position determination unit. The stage has at least two degrees of freedom, and has a movement mechanism for each degree of freedom, and among the movement mechanisms for each degree of freedom, a drive source of the movement mechanism for at least one degree of freedom includes a plurality of piezoelectric elements. Since the piezoelectric element laminate that is stacked and expands and contracts in the stacking direction, the introduction of the introduced substance into the introduced body such as cells can be efficiently and reliably performed with a plurality of injection needles, and the processing efficiency is improved. Is possible.

It is a conceptual diagram of the microinjection apparatus concerning one Embodiment of this invention. It is a top view which shows the arrangement structure of the conveyance means in the microinjection apparatus. It is a partially broken perspective view of one conveyance means in the microinjection apparatus. It is a figure which combines and shows the longitudinal cross-sectional view of an example of the Z-axis moving mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a figure which combines and shows the longitudinal cross-sectional view of the other example of the Z-axis movement mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a figure which combines and shows the horizontal sectional view of the Y-axis moving mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a figure which combines and shows the horizontal sectional view of the X-axis moving mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a figure which combines and shows the horizontal sectional view of the other structural example of the Y-axis moving mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. (A) is a plan view showing a configuration example of a link mechanism in the Y-axis moving mechanism, (B) is a plan view showing another configuration example of the link mechanism and a partially enlarged sectional view thereof, and (C) is the link. It is a top view which shows the other structural example of a mechanism. It is a figure which combines and shows the horizontal sectional view of the X-axis moving mechanism in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a top view which shows the further another structural example of the link mechanism in the X-axis moving mechanism of the conveyance means. It is a perspective view of the petri dish stage in a microinjection apparatus. It is sectional drawing of an example of the cell adsorption substrate unit used for fixation of a floating cell. It is a top view of the XY stage apparatus in a container position adjustment means. (A) is a diagram showing a combination of a horizontal sectional view of another configuration example of the Y-axis moving mechanism in the conveying means and a block diagram of a conceptual configuration of the control system, and (B) is an X-axis moving mechanism in the conveying means. It is a figure which combines and shows the horizontal sectional view of the other structural example, and the block diagram of the conceptual structure of the control system. It is a top view which shows one structural example of the link mechanism of the Y-axis moving mechanism and the X-axis moving mechanism. It is a top view which shows another structural example of the link mechanism. It is a graph showing the effect of another example of composition, and showing the relation between a crankshaft rotation angle and slider movement amount. It is a conceptual diagram of a prior art example.

An embodiment of the present invention will be described with reference to FIGS. FIG. 1 shows a conceptual diagram of this microinjection apparatus. This microinjection apparatus is an apparatus that introduces a substance to be introduced into a body to be introduced by inserting a minute injection needle 11 filled with the substance to be introduced into the body to be introduced. The introducer is, for example, a cell. This cell may be a human body cell or a cell of an organism such as any other animal or plant.
The microinjection apparatus includes a container position adjusting unit 1, a plurality of transfer units 4 which are manipulators individually provided for a plurality of injection needles 11, and the transfer unit 4 between an injection preparation position and a retracted position. It comprises a transporting means retracting mechanism 39 that moves forward and backward, an imaging means 2, a position determining means 3, and a control device 5 that controls the operation of the entire apparatus.
As will be described in detail later, the control device 5 includes a position determination unit 3, a responsible introducer determining unit 87, and an injection operation control unit 88, and the responsible introducer determining unit 87 and the injection operation A control unit 80 is configured by the control unit 88. The injection operation control unit 88 includes a transport unit individual control unit 88 a that controls each transport unit 4.

The container position adjusting means 1 is a means for adjusting the position of the container 12 by moving a container mounting table 13 in which a container 12 such as a petri dish for accommodating an introduction target is held in a horizontal state in two horizontal orthogonal directions. Yes, it is composed of an XY stage device 6. For example, as shown in a plan view in FIG. 14, the XY stage device 6 includes a lower movable table 183 that is installed on a base 181 so as to be movable in the Y-axis direction via a guide 182, and the lower movable table 183. An upper movable base 185 which is movably installed in the X-axis direction via the guide 184, and movable base drive mechanisms 186, 187 for the respective axes for moving the lower movable base 183 and the upper movable base 185 in the movable direction. Consists of. The movable table driving mechanisms 186 and 187 may be ultrasonic motors or linear motors, or may be composed of a motor and a rotation / linear motion conversion mechanism such as a ball screw. The container mounting table 13 shown in FIG. 1 is installed on the upper movable table 185, or the upper movable table 185 itself becomes the container mounting table 13.
Note that the X axis, Y axis, and Z axis in this specification do not represent each axis of rectangular coordinates defined as a common coordinate system in the entire microinjection apparatus, but each freedom in the description of the individual apparatus. Expressed as an axis for degrees.

  Only one imaging means 2 (FIG. 1) is installed at a predetermined position above the container mounting table 13 so as to overlook the container 12 made of a petri dish or the like. The imaging unit 2 is a camera or the like that captures an image such as a planar view image obtained by locally enlarging the inside of the container 12 through the magnifying lens 2 a, and the image is processed by the image processing unit 7.

  The position determination means 3 is a means for determining the position of the introducer from the image obtained by the imaging means 2, and is included as a part of the control device 5, for example. The position determination means 3 determines the position of the introducer by, for example, checking the image processing result by the image processing means 7 with an appropriately set setting reference. For example, the position determination unit 3 determines the position of each introduced object from the positional relationship of each introduced object in the entire image.

  The transport means 4 is a means composed of an XYZ stage device or the like that moves each injection needle 11, and a plurality of such means are provided. As shown in a plan view in FIG. 2, the plurality of transport means 4 are positioned above the container mounting table 13 and are arranged substantially radially with respect to the center of the container 12 so as to surround the container 12. In the example of the figure, a plurality of (for example, three) conveying means 4 are arranged at equal angular intervals on the left and right sides of the container 12, and the left and right conveying means 4 are collinear with respect to the radiation center. Located opposite to each other.

  FIG. 3 is a perspective view of the overall configuration of a transport unit with a retracting mechanism including one transport unit 4 and a transport unit retracting mechanism 39 that supports the transport unit 4. The conveying means 4 has 3 degrees of freedom. The moving mechanism responsible for one degree of freedom moves the injection needle 11 in one direction (X-axis direction) of two directions (X-axis direction and Y-axis direction) perpendicular to the horizontal direction with respect to the container mounting table 13. This is the X-axis moving mechanism 14. Another moving mechanism that bears one degree of freedom is the Y-axis moving mechanism 15 that moves the injection needle 11 in one other direction (Y-axis direction) of two directions orthogonal to each other in the horizontal direction. Still another moving mechanism that bears one degree of freedom is a Z-axis moving mechanism 16 that moves the injection needle 11 in the direction of the injection needle central axis, which is a direction inclined toward the inside of the container 12.

  FIG. 4 shows a configuration example of the Z-axis moving mechanism 16. The Z-axis moving mechanism 16 includes a hollow box-shaped fixing base 17 and a needle support member that is provided so as to protrude upward from the fixing base 17 at one end of the fixing base 17 and that removably supports the injection needle 11. 18 and piezoelectric element laminates 19A, 19B1, and 19B2 disposed in the fixed base 17 and serving as a driving source. The needle support member 18 is configured to detachably support the injection needle 11 and can easily replace the injection needle 11 in order to damage the injection needle 11 or replace the introduced substance. The needle support member 18 has a general T shape having a standing piece portion 18a and a horizontal piece portion 18b, and the horizontal piece portion 18b moves on the fixed base 17 in the protruding direction of the injection needle 11 via the guide mechanism 21. The stand piece 18a is supported on the inwardly facing surface of one end of the fixed base 17 via a spring member 20A made of a leaf spring or the like.

  Each piezoelectric element laminate 19A, 19B1, 19B2 is a laminated piezoelectric element in which a plurality of piezoelectric elements 19a are laminated in the displacement direction to form a rod-like body. Of these piezoelectric element laminates 19A, 19B1 and 19B2, the two piezoelectric element laminates 19B1 and 19B2 are arranged in a straight line and connected to each other in series by a fastening member 47. The laminated body 19B is obtained. The piezoelectric element laminate 19B and the remaining one set of piezoelectric element laminates 19A are arranged in parallel vertically so as to be parallel to each other along the lamination direction, and these two sets of piezoelectric element laminates 19A and 19B are arranged in parallel. They are connected in series via the fastening member 48. The fastening member 48 includes a longitudinal direction portion 48a disposed between the upper and lower piezoelectric element laminates 19A and 19B in parallel with the piezoelectric element laminates 19A and 19B, and the vertical direction at both ends of the longitudinal direction portion 48a. Are generally Z-shaped with protrusions 48b and 48c projecting in opposite directions. One set of piezoelectric element laminates 19 </ b> A has one end connected to a protrusion 48 b on one end side of the fixing base 17 of the fastening member 48, and the other end supported by the other end of the fixing base 17. Between the protrusion 48b of the fastening member 48 and one end of the fixed base 17, a spring member 20B such as a leaf spring that preloads the piezoelectric element laminate 19A is interposed. One end of the piezoelectric element laminate 19B, that is, the end opposite to the connecting portion by the fastening member 47 in one piezoelectric element laminate 19B1 constituting the piezoelectric element laminate 19B is a standing piece portion of the needle support member 18. 18a. Further, the other end of the piezoelectric element laminate 19B, that is, the end of the other piezoelectric element laminate 19B2 constituting the piezoelectric element laminate 19B opposite to the connecting portion by the fastening member 47 is the fixed base 17 The other end side is connected to the protrusion 48c of the fastening member 48. The piezoelectric element laminate 19 </ b> B is preloaded by a spring member 20 </ b> A such as a leaf spring interposed between the standing piece 18 a of the needle support member 18 and the other end of the fixed base 17. Thereby, the needle support member 18 can advance and retract in the protruding direction of the injection needle 11 by the displacement in the stacking direction of the two sets of piezoelectric element stacks 19A and 19B.

Of the piezoelectric element laminates 19A and 19B, the piezoelectric element laminate 19A and one piezoelectric element laminate 19B2 in the piezoelectric element laminate 19B are used for positioning the needle support member 18, that is, for positioning the injection needle 11. Used as a driving source. That is, these piezoelectric element laminates 19A and 19B2 are displaced by a positioning signal that is a voltage applied from the Z-axis (insertion direction) position control unit 51.
Of the piezoelectric element laminates 19A and 19B, the piezoelectric element laminate 19B1 directly connected to the needle support member 18 in the piezoelectric element laminate 19B is a drive source for causing the needle support member 18 to vibrate in the insertion direction of the injection needle 11. Used as. The displacement of the piezoelectric element laminate 19 </ b> B <b> 1 for applying vibration is repeatedly changed by a vibration driving signal that is a voltage applied from the vibration driving means 54.

The Z-axis position control unit 51 receives a position command from the position command unit 52 to the voltage generator 53, and applies a voltage corresponding to the piezoelectric element laminates 19A and 19B2 from the voltage generator 53 based on the position command. . The Z-axis position control unit 51 is provided in the Z-axis transport means individual control unit 88a in the injection operation control unit 88 of FIG. Further, the position command unit 52 uses the target position determined by the assigned body determining unit 87 as the position command based on the position information obtained by the position determination unit 3 of FIG. The position command unit 52 may be provided as a part of the assigned introducer determining unit 87.
The vibration drive means 54 applies an alternating voltage having a predetermined frequency to the piezoelectric element laminate 19B1 as the vibration drive signal from the voltage generator 56. The frequency of the alternating voltage, that is, the frequency of vibration applied to the needle support member 18 can be switched by a command from the frequency variable means 55 to the voltage generator 56.

  FIG. 5 shows another configuration example of the Z-axis moving mechanism 16. In this configuration example, in the configuration example of FIG. 4, a position command unit 57 that gives a position command to the voltage generator 56 based on the position information obtained by the position determination unit 3 in FIG. ing. As a result, the vibration driving means 54 applies a voltage obtained by superimposing the positioning signal for positioning the needle support member 18 and the vibration driving signal to the piezoelectric element laminate 19B1. The position command unit 57 may be provided as a part of the assigned introducer determination unit 87 in FIG. Other configurations are the same as those in the configuration example of FIG.

  This Z-axis moving mechanism 16 incorporates a sensor (not shown) for measuring the relative displacement between the fixed base 17 and the needle support member 18. Examples of the sensor include a strain sensor that detects the strain of the spring members 20A and 20B such as a leaf spring that applies a preload to the piezoelectric element laminates 19A and 19B, and a static that measures the gap between the fixing base 17 and the needle support member 18. A capacitance sensor, a magnetic sensor, an optical sensor, or the like can be used.

  As shown in FIG. 3, the Z-axis moving mechanism 16 is fixed to the inclined mounting base 22a at one end of the upper movable support 22 so that the injection angle of the injection needle 11 becomes a predetermined angle inclined downward. Yes. Note that the Z-axis moving mechanism 16 may include an angle variable mechanism (not shown) that freely changes the injection angle of the injection needle 11 with respect to the upper movable support 22. The upper movable support 22 has a rectangular frame shape with one side open. The upper movable support 22 is supported on a flat lower movable support 23 via a pair of left and right guide mechanisms 24 so as to be movable in one horizontal direction (Y-axis direction). On the lower movable support 23, the Y-axis moving mechanism 15 is installed. The lower movable support body 23 is supported on a flat plate-like transport means base 28 disposed therebelow via a guide mechanism 29 so as to be movable in the X-axis direction. The X-axis moving mechanism 14 is installed below the conveying means base 28.

  The Y-axis moving mechanism 15 is shown in a horizontal sectional view in FIG. The Y-axis moving mechanism 15 includes a fixed base 25, a movable piece 26, and two sets of piezoelectric element laminates 19C and 19D serving as a drive source. The fixing base 25 includes a main frame portion 25a extending in the X-axis direction, a pair of side frame portions 25b and 25d extending in the width direction (Y-axis direction) from both ends of the main frame portion 25a, and a front end of the side frame portion 25b. A horizontal cross section extending in the X-axis direction in parallel with the main frame portion 25a has a hollow box shape having an L-shaped sub frame portion 25c. The movable piece 26 extends from the front end of the side frame portion 25b at one end of the fixed base 25 toward the side frame portion 25d at the other end, and has a horizontal section as an L-shaped movable piece 26. The movable piece 26 serves as an enlargement mechanism that enlarges the displacement of the piezoelectric element laminates 19C and 19D, and is formed of an elastic material such as metal or synthetic resin together with the fixed base 25. The movable piece 26 includes a main frame parallel piece portion 26a extending substantially parallel to the main frame portion 25a from the side frame portion 25b at one end of the fixed stand 25, and the other end of the fixed stand 25 from the other end of the main frame parallel piece portion 26a. The side frame parallel piece portion 26b extends in parallel to the side frame portion 25d on the inner side of the side frame portion 25d. The connecting portion between the side frame portion 25b of the fixed base 25 and the main frame parallel piece portion 26a of the movable piece 26 and the connecting portion between the main frame parallel piece portion 26a and the side frame parallel piece portion 26b of the movable piece 26 are thin portions 26c. It is said that. Further, the middle portion in the longitudinal direction of the main frame parallel piece portion 26a of the movable piece 26 is also a thin portion 26d. As a result, the side frame parallel piece portion 26b of the movable piece 26 can be swung so as to be bent with the thin-walled portion 26c at the base end as a swing center. Further, the main frame parallel piece 26a of the movable piece 26 is bent at the thin-walled portion 26d at the middle portion in the longitudinal direction, and the middle portion in the direction perpendicular to the longitudinal direction (Y-axis direction) due to increase or decrease in the bending angle. Can be moved forward and backward.

  The two sets of piezoelectric element laminates 19C and 19D are both laminated piezoelectric elements, and are arranged in parallel in the front-rear direction so as to be parallel to the main frame portion 25a of the fixed base 25 along the lamination direction. These two sets of piezoelectric element laminates 19 </ b> C and 19 </ b> D are connected in series via a fastening member 58. The fastening member 58 includes a longitudinal portion 58a disposed in parallel with both the piezoelectric element laminates 19C and 19D between the front and rear piezoelectric element laminates 19C and 19D, and a longitudinal direction at both ends of the longitudinal direction portion 58a. Are generally Z-shaped having protrusions 58b and 58c protruding in opposite directions. One set of the piezoelectric element laminate 19C is supported at one end by the side frame 25b of the fixed base 25, and the other end is connected to the protrusion 58b of the fastening member 58 facing the fixed base side frame 25d. . The other set of piezoelectric element laminates 19D has one end connected to the projection 58c of the fastening member 58 facing the fixed base side frame 25b and the other end connected to the side frame parallel piece of the movable piece 26. 26b. A spring member 27A such as a leaf spring for applying a preload to the piezoelectric element laminate 19C is provided between the side portion 25ca extending in the width direction at the front end of the sub-frame portion 25c of the fixing base 25 and the protruding portion 58b of the fastening member 58. Intervene. Between the side frame portion 25d of the fixed base 25 and the side frame parallel piece portion 26b of the movable piece 26, a spring member 27B such as a leaf spring for applying a preload to the piezoelectric element laminate 19D is interposed. Further, a preload is applied to the side frame parallel piece portion 26b of the movable piece 26 between the side portion 25ca extending in the width direction of the tip of the sub frame portion 25c of the fixed base 25 and the side frame parallel piece portion 26b of the movable piece 26. A spring member 27C such as a leaf spring is provided. As a result, the side frame parallel piece 26b of the movable piece 26 swings due to the displacement caused by expansion and contraction in the stacking direction of the two sets of piezoelectric element laminates 19C and 19D, and the swing displacement is expanded to expand the main frame parallel piece. The displacement of the portion 26a in the Y-axis direction is obtained. This displacement is transmitted to the upper movable support 22 (FIG. 3) that supports the Z-axis moving mechanism 16, whereby the injection needle 11 can move in the Y-axis direction.

  The two sets of piezoelectric element laminates 19 </ b> C and 19 </ b> D are displaced by a positioning signal that is a voltage applied from the Y-axis position control unit 59. The Y-axis position control unit 59 receives a position command from the position command unit 60 to the voltage generator 61, and applies a voltage corresponding to the piezoelectric element stacks 19C and 19D from the voltage generator 61 based on the position command. To do. The Y-axis position control unit 59 is provided in the Y-axis transport means individual control unit 88a in the injection operation control unit 88 of FIG. Further, the position command unit 61 uses the target position determined by the assigned introducer determination unit 87 as the position command based on the position information obtained by the position determination unit 3 of FIG. The position command unit 61 may be provided as a part of the assigned introducer determining unit 87.

  In the case of the Y-axis moving mechanism 15, a sensor (not shown) for measuring the relative displacement between the fixed base 25 and the movable piece 26 is incorporated. Examples of the sensor include a strain sensor that detects a strain of the leaf springs 27A and 27B that applies a preload to the piezoelectric element laminates 19C and 19D, a capacitance sensor that measures a gap between the fixed base 25 and the movable piece 26, and a magnetic sensor. A sensor, an optical sensor, or the like can be used.

  FIG. 7 shows the X-axis moving mechanism 14 in a horizontal sectional view. The X-axis moving mechanism 14 includes a fixed base 35, a movable piece 36, and two sets of piezoelectric element laminates 19E and 19F serving as driving sources. The fixed base 35 includes a main frame portion 35a extending in the X-axis direction, a pair of side frame portions 35b and 35d extending in the width direction (Y-axis direction) from both ends of the main frame portion 35a, and a front end of the side frame portion 35b. A horizontal box extending in the X-axis direction in parallel with the main frame portion 35a has a hollow box shape having an L-shaped sub-frame portion 35c. The movable piece 36 extends from the tip of the side frame portion 35b at one end of the fixed base 35 toward the side frame portion 35d at the other end, and has a horizontal cross section in an L shape. The movable piece 36 serves as a first enlargement mechanism that enlarges the displacement of the piezoelectric element laminates 19E and 19F, and is integrally formed of an elastic material such as metal or synthetic resin with the fixed base 35. The movable piece 36 includes a main frame parallel piece portion 36a extending substantially parallel to the main frame portion 35a from the side frame portion 35b at one end of the fixed table 35, and the other end of the fixed frame 35 from the other end of the main frame parallel piece portion 36a. A side frame parallel piece portion 36b extending in parallel with the side frame portion 35d is formed inside the side frame portion 35d. The connecting portion between the side frame portion 35b of the fixed base 35 and the main frame parallel piece portion 36a of the movable piece 36 and the connecting portion between the main frame parallel piece portion 36a and the side frame parallel piece portion 36b of the movable piece 36 are thin portions 36c. It is said that. Further, the intermediate portion in the longitudinal direction of the main frame parallel piece portion 36a of the movable piece 36 is also a thin portion 36d. Thereby, the side frame parallel piece portion 36b of the movable piece 36 can be swung with the thin-walled portion 36c at the base end as a swing center. Further, the main frame parallel piece portion 36a of the movable piece 36 is bent at a thin portion 36d in the middle portion in the longitudinal direction and can swing in a direction perpendicular to the longitudinal direction (Y-axis direction). The configuration so far is the same as that of the Y-axis moving mechanism 15 in FIG.

  Further, a portion which is a half portion closer to the side frame parallel piece portion 36b than the thin portion 36d in the longitudinal direction intermediate portion of the main frame parallel piece portion 36a of the movable piece 36 extends toward the side frame portion 35d side of the fixed base 35. A movable frame portion 37 having an inverted L-shaped horizontal cross section connected to the vicinity of the base end of the side frame portion 35d is integrally formed. The movable frame portion 37 serves as a second expansion mechanism that expands the displacement of the piezoelectric element laminates 19E and 19FC, and includes a thick frame portion 37a substantially parallel to the main frame parallel piece portion 36a of the movable piece 36. The thin frame portion 37b extends from the thick frame portion 37a to the outside of the side frame portion 35d of the fixed base 35 in parallel with the side frame portion 35d. The connecting portion between the thick frame portion 37a and the thin frame portion 37b of the movable frame portion 37 and the connecting portion between the thin frame portion 37b and the vicinity of the base end of the side frame portion 35b of the fixed base 35 are more than the thin frame portion 37b. Furthermore, it is set as the thin thin part 37c. Further, the intermediate portion in the longitudinal direction of the thin frame portion 37b is also a thinner thin portion 37d. Thereby, the thin frame portion 37b of the movable frame portion 37 is bent at the thin portion 37d of the middle portion in the longitudinal direction so that the intermediate portion can advance and retreat in a direction (X-axis direction) perpendicular to the longitudinal direction.

  As in the case of the Y-axis moving mechanism 15 in FIG. 6, the two sets of piezoelectric element laminates 19E and 19F are both laminated piezoelectric elements, and are parallel to the main frame portion 35a of the fixed base 35 along the lamination direction. It is arranged in parallel so as to become. The two sets of piezoelectric element laminates 19E and 19F are connected in series via a fastening member 62. The fastening member 62 includes a longitudinal direction portion 62a disposed between the front and rear piezoelectric element laminates 19E and 19F in parallel with the piezoelectric element laminates 19E and 19F, and a longitudinal direction at both ends of the longitudinal direction portion 62a. And the projections 62b and 62c projecting so as to be opposite to each other. One set of the piezoelectric element laminates 19E is supported by the side frame portion 35b of the fixed base 35, and the other end is connected to the protrusion 62b of the fastening member 62 facing the fixed base side frame portion 35d. . The other set of piezoelectric element laminates 19F has one end connected to the protrusion 62c of the fastening member 62 facing the fixed base side frame 35b and the other end connected to the side frame parallel piece of the movable piece 36. 36b. A spring member 38A such as a leaf spring for applying a preload to the piezoelectric element laminate 19E is provided between the side portion 35ca extending in the width direction at the tip of the sub-frame portion 35c of the fixing base 35 and the protrusion 62b of the fastening member 62. Intervene. Between the side frame portion 35d of the fixed base 35 and the side frame parallel piece portion 36b of the movable piece 36, a spring member 38B such as a leaf spring for applying a preload to the piezoelectric element laminate 19F is interposed. Further, a preload is applied to the side frame parallel piece portion 36b of the movable piece 36 between the side portion 35ca extending in the width direction of the tip of the sub frame portion 35c of the fixed base 35 and the side frame parallel piece portion 36b of the movable piece 36. A spring member 38C such as a leaf spring is provided. As a result, the side frame parallel piece portion 36b of the movable piece 36 is swung by the displacement due to expansion and contraction in the stacking direction of the two sets of piezoelectric element laminates 19E and 19F, and the swing displacement is enlarged to enlarge the main frame parallel piece. This is the displacement of the portion 36a in the Y-axis direction. This displacement is further enlarged and becomes a displacement in the X-axis direction of the thin frame portion 37 b in the movable frame portion 37. This displacement in the X-axis direction is transmitted to the lower movable support body 23 in FIG. 3, whereby the injection needle 11 can move in the X-axis direction.

  The two sets of piezoelectric element laminates 19E and 19F are displaced by a positioning signal that is a voltage applied from the X-axis position control unit 63. The X-axis position control unit 63 receives a position command from the position command unit 64 to the voltage generator 65, and applies a voltage corresponding to the piezoelectric element stacks 19E and 19F from the voltage generator 65 based on the position command. Is done. The X-axis position control unit 63 is provided in the X-axis transport means individual control unit 88a in the injection operation control unit 88 of FIG. Further, the position command unit 64 uses the target position determined by the assigned introducer determining unit 87 based on the position information obtained by the position determination unit 3 of FIG. 1 as the position command. The position command unit 64 may be provided as a part of the assigned introducer determining unit 87.

  In the case of the X-axis moving mechanism 14, a sensor (not shown) for measuring the relative displacement between the fixed base 35 and the movable piece 36 is incorporated. Examples of the sensor include a strain sensor that detects a strain of the leaf springs 38A and 38B that applies a preload to the piezoelectric element laminates 19E and 19F, a capacitance sensor that measures a gap between the fixed base 35 and the movable piece 36, and a magnetic sensor. A sensor, an optical sensor, or the like can be used.

  FIG. 8 shows another configuration example of the Y-axis moving mechanism 15. As in the configuration example of FIG. 6, this configuration example also includes a fixed base 73 and two sets of piezoelectric element laminates 19G and 19H serving as a drive source. The fixed base 73 includes a main frame portion 73a extending in the X-axis direction and a pair of side frame portions 73b and 73c extending in the width direction (Y-axis direction) from both ends of the main frame portion 73a. On the side frame portion 73b at one end of the fixed base 73, an expansion / contraction direction moving body 74 facing the side frame portion 73c at the other end is supported via a spring member 27D so as to be movable in the X-axis direction.

  The two sets of piezoelectric element laminates 19G and 19H are both laminated piezoelectric elements, and are arranged in parallel in the front-rear direction so as to be parallel to the main frame portion 73a of the fixed base 73 along the lamination direction. These two sets of piezoelectric element laminates 19G and 19H are connected in series via a fastening member 78. The fastening member 78 includes a longitudinal direction portion 78a arranged in parallel with both the piezoelectric element laminates 19G and 19H between the front and rear piezoelectric element laminates 19G and 19H, and a longitudinal direction at both ends of the longitudinal direction portion 78a. And the projections 78b and 78c projecting so as to be opposite to each other. One set of the piezoelectric element laminates 19G is supported at one end by the side frame portion 73c of the fixed base 73, and the other end is connected to the protrusion 78b of the fastening member 78 facing the movable body 74 in the telescopic direction. The other set of piezoelectric element laminates 19H has one end connected to the protrusion 78c of the fastening member 78 facing the fixed base side frame 73c and the other end connected to the telescopic moving body 74. Yes. The spring member 27D applies a preload to the piezoelectric element laminate 19H. Thereby, the expansion-contraction direction moving body 74 can be displaced in the X-axis direction in accordance with the expansion / contraction displacement of the piezoelectric element laminates 19G, 19H.

  In the Y-axis moving mechanism 15, a link mechanism 75 is provided as an expansion mechanism that expands and contracts the piezoelectric element stacks 19G and 19H to displacement in a direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction). . The link mechanism 75 is connected to the fixed base side frame 73c by one fixed joint 76, and is connected to the telescopic direction moving body 74 by one movable joint 77A.

FIG. 9 shows various configuration examples of the link mechanism 75. In the configuration example of FIG. 9A, a link mechanism 75 is configured by one fixed joint 76, two movable joints 77A and 77B, and three links 81A, 81B, and 81C.
Each of the fixed joints 76 and the movable joints 77A and 77B is a joint that constitutes a rotatable node, and the center of rotation is the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19G and 19H and the expansion / contraction thereof. It is perpendicular to any direction (Y-axis direction) orthogonal to the direction (X-axis direction). The same applies to the joints 76, 77A, 77B, and 77C in FIGS. 9B and 9C.
One end of each of the first and second links 81A and 81B is connected to the fixed joint 76 and the first movable joint 77A, and the other end is connected to each other by a second movable joint 77B. The third link 81C has one end connected to the second movable joint 77B, and the other end movable only in the orthogonal direction (Y-axis direction), such as a linear guide, a linear motion bearing, or the like (see FIG. It becomes the movable part 82 restrained by (not shown). The fixed joint 76 is fixed in position with respect to the side frame portion 73c of the fixed base 73 to which the fixed ends of the piezoelectric element laminates 19G and 19H are fixed. The guide mechanism for guiding the movable portion 82 is provided on the fixed base 73. The first movable joint 77A is provided in an expansion / contraction direction moving body 74 that can move integrally with the expansion / contraction side ends of the piezoelectric element laminates 19G and 19H, and is movable together with the expansion / contraction direction moving body 74. The movable portion 82 of the third link 81 </ b> C serves as a displacement expansion output portion of the link mechanism 75.

According to this configuration, the first movable joint 77A is displaced in the X-axis direction due to the expansion and contraction of the piezoelectric element laminates 19G and 19H, and this displacement is enlarged so that the second movable joint 77B is displaced in the Y-axis direction. Accordingly, the third link 81C changes the rotation angle, so that the movable portion 82 at the other end is guided in the Y-axis direction by being guided by the guide mechanism such as the linear guide.
In this case, by using a rolling bearing for the fixed joint 76 and the movable joints 77A and 77B, the frictional resistance can be reduced, and by providing an appropriate preload to the rolling bearing, rattling can be suppressed and precise positioning can be realized. Moreover, since there is no elastic deformation part, design is easy.

In the configuration example of FIG. 9B, a link mechanism 75 is configured by one fixed joint 76, three movable joints 77A, 77B, and 77C, and three links 81A, 81B, and 81C. In this example, in the configuration example of FIG. 9A, the movable joint that connects the base end of the third link 81C is the third movable joint 77C that is located at a different position from the second movable joint 77B. It is.
That is, the other end of the first link 81A having the base end connected to the fixed joint 76 is connected to the second movable joint 77B in the middle of the second link 81B having the base end connected to the first movable joint 77A. Connect through. The third link 81C is connected to the tip of the second link 81B via the third movable joint 77C. A movable portion 82 constrained by a guide mechanism (not shown) similar to the above so that the tip of the third link 81 is movable only in a direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction); To do. The fixed joint 76 and the first movable joint 77A are provided on the fixed base 73 and the expansion / contraction direction moving body 74, respectively, as in the example of FIG. 9A. In the example of FIG. 9B, the movable portion 82 at the tip of the third link 81 </ b> C serves as a displacement expansion output portion of the link mechanism 75.
The second movable joint 77B has, for example, a bearing 122 attached to a connecting pin 121 fixed to one of the first or second links 81A and 81B, as shown in an enlarged cross-sectional view in FIG. Via the other of the first or second link 81A, 81B. The bearing 122 is fitted and attached to a hole provided in one of the links 81A and 81B. The bearing 122 may be either a rolling bearing such as a ball bearing or a sliding bearing. For example, the bearing 122 is a preloadable rolling bearing.

According to this configuration, the movable joint 77A is displaced in the X-axis direction due to the expansion and contraction of the piezoelectric element laminates 19G and 19H, and this displacement is enlarged so that another movable joint 77C is displaced in the Y-axis direction. When the link 81C changes the rotation angle, the movable portion 82 at the tip thereof is guided by the guide mechanism and displaced in the Y-axis direction.
In the configuration examples of FIGS. 9A and 9B, the angle of the third link 81C can be varied independently even if the size error of each link 81C, 81B, 81C or the influence of thermal deformation occurs. The movable part 82 which is absorbed by being present and becomes a displacement expansion output part can move in the linear direction in the Y-axis direction along the guide mechanism such as the linear guide.
In the configuration example of FIG. 9A, since the three links 81A, 81B, 81C are connected at the position of the second movable joint 77B, it is necessary to provide two bearings side by side in the axial direction thereof. The dimension in the thickness direction increases. However, in the case of the configuration example in FIG. 9B, the movable joints 77B and 77C only need to be provided with one bearing 122 in the axial direction, and the thickness dimension is larger than that in the configuration example in FIG. It can be reduced to 1/2.

In the configuration example of FIG. 9C, a link mechanism 75 is configured by one fixed joint 76, two movable joints 77A and 77B, and two links 81A and 81B.
That is, the other end of the first link 81A having the base end connected to the fixed joint 76 is connected to the second movable joint 77B in the middle of the second link 81B having the base end connected to the first movable joint 77A. Connect through. 7A and 7B, the fixed joint 76 is connected to the side frame portion 73c of the fixed base 73, and the first movable joint 77A is connected to the telescopic moving body 74. In this configuration example, the movable portion 82 at the tip of the second link serves as a displacement expansion output portion of the link mechanism 75.
In the case of this configuration, the length of the second link 81B is twice that of the first link 81A, and the second movable joint 77B is disposed at the center position of the link 81B. The moving direction is constrained without providing a guide mechanism such as a linear guide. The moving direction is a direction perpendicular to the straight line connecting the centers of the fixed joint 76 and the first movable joint 77A, that is, a direction orthogonal to the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19G and 19H (Y It can be displaced in the axial direction).

  According to this configuration, the movable joint 77A is displaced in the X-axis direction due to the expansion and contraction of the piezoelectric element laminates 19G and 19H, and this displacement is expanded to displace the movable part 82 that is the tip of the second link 81B in the Y-axis direction. To do. This configuration example is the most compact. Also in this example, the movable joint 77B only needs to be provided with one bearing in the axial direction, and the thickness dimension can be reduced to ½ compared to the configuration example of FIG. 9A.

  In the Y-axis moving mechanism 15 of FIG. 8, the link mechanism 75 of the example of FIG. 9C is provided as an enlargement mechanism. Other configurations are the same as those of the Y-axis moving mechanism 15 shown in FIG.

  FIG. 10 shows another configuration example of the X-axis moving mechanism 14. Also in this configuration example, as in the case of the Y-axis moving mechanism 15 in FIG. 8, the fixed base 83 and two sets of piezoelectric element laminates 19I and 19J serving as a drive source are provided. The fixed base 83 includes a main frame portion 83a extending in the X-axis direction and a pair of side frame portions 83b and 83c extending in the width direction (Y-axis direction) from both ends of the main frame portion 83a. A movable body 84 facing the side frame 83c at the other end is supported by the side frame 83b at one end of the fixed base 83 so as to be movable in the X-axis direction via a spring member 27E.

  The two sets of piezoelectric element laminates 19I and 19J are both laminated piezoelectric elements, and are arranged in parallel in the front-rear direction so as to be parallel to the main frame portion 83a of the fixed base 83 along the lamination direction. These two sets of piezoelectric element laminates 19 </ b> I and 19 </ b> J are connected in series via a fastening member 85. The fastening member 85 includes a longitudinal direction portion 85a disposed in parallel with both the piezoelectric element laminates 19I and 19J between the front and rear piezoelectric element laminates 19I and 19J, and a longitudinal direction at both ends of the longitudinal direction portion 85a. Are generally Z-shaped having protrusions 85b and 85c protruding in opposite directions. One set of the piezoelectric element laminates 19I is supported at one end by the side frame 83c of the fixed base 83, and the other end is connected to the protrusion 85b of the fastening member 85 facing the movable body 84 side. The other set of piezoelectric element laminates 19 </ b> J has one end connected to the protrusion 85 c of the fastening member 85 facing the fixed base frame 83 c and the other end connected to the movable body 84. The spring member 27E applies a preload to the piezoelectric element laminate 19J. Thereby, the movable body 84 can be displaced in the X-axis direction in accordance with the expansion and contraction of the piezoelectric element laminates 19I and 19J.

  In the X-axis moving mechanism 14, the first link is a first expansion mechanism that expands the expansion and contraction of the piezoelectric element stacks 19 </ b> I and 19 </ b> J to a displacement (Y-axis direction) perpendicular to the expansion / contraction direction (X-axis direction). A second link is provided as a second expansion mechanism that provides the mechanism 101 and expands the displacement expanded by the first expansion mechanism (first link mechanism 101) to the displacement in the expansion / contraction direction of the piezoelectric element stacks 19I and 19J. A mechanism 102 is provided. As these both link mechanisms 101 and 102, the thing of the same structure as the link mechanism 75 shown in FIG.9 (C) is used here. Note that the second link mechanism 102 is in a posture that is 90-degree changed with respect to the first link mechanism 101.

  That is, the first link mechanism 101 includes one fixed joint 106, two movable joints 107A and 107B, and two links 108A and 108B. Specifically, one end is displaced in the expansion / contraction direction according to expansion / contraction of the first link 108A connected to the side frame portion 83c of the fixing base 83 by one fixed joint 106 and the piezoelectric element laminates 19I and 19J. One end is connected to the movable body 84 by one movable joint 107A, and the middle part is constituted by the second link 108B connected to the other end of the first link 108A by another one movable joint 107B. The other end of the second link 108B can be displaced in a direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction) of the piezoelectric element stacks 19I and 19J.

  The second link mechanism 102 is also configured by one fixed joint 116, two movable joints 117A and 117B, and two links 118A and 118B. Specifically, the first link 118A, one end of which is connected to the side frame 83c of the fixed base 83 by one fixed joint 116, and the expansion / contraction direction of the piezoelectric element laminates 19I and 19J are orthogonal to each other. One end is connected to the other end of the second link 108B of the first link mechanism 101 that is displaced in the Y-axis direction by one movable joint 117A, and the middle portion is connected to the other end of the first link 118A by another one The second link 118B is connected to the movable joint 117B. Thus, the first and second link mechanisms 101 and 102 are combined in two stages via the movable joint 117A on the same plane. In this case, the distal end of the second link 118B in the second link mechanism 102 becomes a movable portion 119 that is a displacement expansion output portion, and can be displaced in the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19I and 19J. The

  According to this configuration, in the first link mechanism 101, the movable joint 107A is displaced in the X-axis direction by the expansion and contraction of the piezoelectric element laminates 19I and 19J, and this displacement is enlarged to expand the movable joint 117A that is the other end of the link 10B. Is displaced in the Y-axis direction. In the second link mechanism 102, the displacement of the movable joint 117A in the Y-axis direction is enlarged, and the movable portion 119, which is the other end of the link 118B, expands and contracts in the piezoelectric element stacks 19I and 19J (X-axis direction). It is displaced to. Other configurations are the same as those of the X-axis moving mechanism 14 shown in FIG.

  In the X-axis moving mechanism 14 in FIG. 10, the first and second link mechanisms 101 and 102 are combined in two stages via the movable joint 117A with the configuration example shown in FIG. 9C. However, the present invention is not limited to this, and the configuration example shown in FIG. 9B may be combined in two stages via a movable joint. An example is shown in FIG. In this example, the second link mechanism 102 is turned 90 degrees with respect to the first link mechanism 101.

  In the example of FIG. 11, the first link mechanism 101 includes one fixed joint 106, three movable joints 107A, 107B, and 107C, and two links 108A and 108B. The second link mechanism 102 has one fixed joint 116, three movable joints 117A, 117B, and 117C, and two links 118A and 118B. By connecting the movable portion 82 in the first link mechanism 101 and the base end of the second link 118B in the second link mechanism 102 with a movable joint 117A, the first and second link mechanisms on the same plane. 101 and 102 are combined in two stages. The tip of the second link 118B of the second link mechanism 102 becomes a movable part 119 which is a displacement expansion output part, and can be freely displaced in the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19I and 19J.

  As another configuration example of the link mechanism 75, as shown in FIG. 15, the link mechanism 75 is a crank-slider mechanism comprising one fixed joint 76, two movable joints 77A and 77B, and two links 81A and 81B. This crank-slider mechanism is capable of causing displacement in the extending direction of the piezoelectric element to be arbitrary on the circumference of the fixed joint 76 via the two movable joints 77A and 77B and the two links 81A and 81B. It is also possible to change the direction and further expand the displacement.

  FIG. 15A shows another configuration example of the Y-axis moving mechanism 15. Similarly to the configuration example of FIG. 6, the Y-axis moving mechanism 15 according to the other configuration example also includes a fixed base 73 and two sets of piezoelectric element laminates 19G and 19H serving as a drive source. The fixed base 73 includes a pair of main frame portions 73a and 73b extending in the X-axis direction, a pair of side frame portions 73c and 73d extending in the width direction (Y-axis direction) from both ends of the main frame portions 73a and 73b, and 4 And a side plate 73e for fastening one frame portion. On the side frame portion 73c at one end of the fixed base 73, an expansion / contraction direction moving body 74 facing the side frame portion 73d at the other end is supported via a spring member 27D so as to be movable in the X-axis direction.

  The two sets of piezoelectric element laminates 19G and 19H are both laminated piezoelectric elements, and are arranged in parallel in the front-rear direction so as to be parallel to the main frame portions 73a and 73b of the fixed base 73 along the lamination direction. . These two sets of piezoelectric element laminates 19G and 19H are connected in series via a fastening member 78. The fastening member 78 includes a longitudinal direction portion 78a arranged in parallel with both the piezoelectric element laminates 19G and 19H between the front and rear piezoelectric element laminates 19G and 19H, and a longitudinal direction at both ends of the longitudinal direction portion 78a. And the projections 78b and 78c projecting so as to be opposite to each other. One set of the piezoelectric element laminates 19G is supported at one end by the side frame portion 73c of the fixed base 73, and the other end is connected to the protrusion 78b of the fastening member 78 facing the movable body 74 in the telescopic direction. The other set of piezoelectric element laminates 19H has one end connected to the protrusion 78c of the fastening member 78 facing the fixed base side frame 73d side and the other end connected to the telescopic moving body 74. Yes. The spring member 27D applies a preload to the piezoelectric element laminate 19H. Thereby, the expansion-contraction direction moving body 74 can be displaced in the X-axis direction in accordance with the expansion / contraction displacement of the piezoelectric element laminates 19G, 19H.

In the Y-axis moving mechanism 15, a link mechanism 75 is provided as an expansion mechanism that expands and contracts the piezoelectric element stacks 19G and 19H to displacement in a direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction). . The link mechanism 75 is connected to the fixed base side frame 73d by one fixed joint 76, and is connected to the telescopic direction moving body 74 by one movable joint 77A.
According to this configuration, the movable joint 77A is displaced in the X-axis direction due to the expansion and contraction of the piezoelectric element laminates 19G and 19H, and the movable joint 77B rotates about the fixed joint 76. Therefore, the movable part 82 fixed to the first link 81A is displaced in the Y-axis direction. Therefore, by setting the output portion at an arbitrary position on the circumference of the fixed joint 76, the displacement can be expanded in an arbitrary direction. Therefore, the number of parts of the Y-axis moving mechanism 15 can be reduced and the size can be reduced.

  FIG. 15B shows another configuration example of the X-axis moving mechanism 14. The X-axis moving mechanism 14 in FIG. 15B also has the same configuration as that in FIG. 15A. By changing the positions of the movable joint 77B and the movable portion 82 fixed to the link 81A, the X-axis moving mechanism 14 is changed in the X-axis direction. Displace. Therefore, by setting the output portion at an arbitrary position on the circumference of the fixed joint 76, the displacement can be increased in an arbitrary direction. Therefore, the number of parts of the Y-axis moving mechanism 15 can be reduced and the size can be reduced. It becomes possible.

  FIG. 16 shows an enlarged configuration example of the link mechanism 75 shown in FIGS. 15 (A) and 15 (B). In the configuration example of FIG. 15, one fixed joint 76, two movable joints 77A and 77B, and two links 81A and 81B constitute a link mechanism 75 serving as a crank / slider mechanism. The joints 77A and 77B constitute joints that can rotate freely, and the center of rotation is the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19G and 19H and the expansion / contraction direction (X-axis direction). It is perpendicular to any of the directions orthogonal to the Y axis direction (Y-axis direction).

  One end of each of the first and second links 81A and 81B is connected to the fixed joint 76 and the first movable joint 77A, and the other end is connected to each other by a second movable joint 77B. The first movable joint 77A is provided in an expansion / contraction direction moving body 74 that can move integrally with the expansion / contraction side ends of the piezoelectric element laminates 19G and 19H, and is movable together with the expansion / contraction direction moving body 74.

  FIG. 17 shows a configuration of an offset crank mechanism in which the fixed joint 76 is not on the extension line in the sliding direction of the first movable joint 77A and has an offset. The effect of the offset will be described with reference to FIG. FIG. 18 shows the amount of movement of the first movable joint 77A when the link 81A is rotated once around the fixed joint 76. FIG. The solid line in FIG. 18 shows the case where there is no offset. When the movable joint 77A reciprocates, the link 81A rotates around the fixed joint 76, and the rotation angle of the movable joint 77A moves to the left in the drawing as the maximum. The rotation angle is 180 degrees, and the relationship between the rotation angle of the link 81A and the amount of movement of the movable joint 77A is about 180 degrees. On the other hand, the broken line in FIG. 18 shows the case where the offset δ shown in FIG. 17 is present, and the rotational angle of the link 81A with respect to the reciprocating motion of the movable joint 77A is the case where the movable joint 77B is on the offset side (upper side in the figure). Will be bigger.

  When the displacement of the moving amount of several hundred μm of the movable joint 77A is expanded to several mm by the movable portion 82 as in the present invention, the rotation angle of the link 81A is about several degrees. In general, the displacement enlargement ratio is determined by the lengths of the first and second links 81A and 81B, the initial position of the movable joint 77A, and the lengths of the movable portion 82 and the fixed joint 76. If the offset δ is provided as in the present invention and the rotation angle of the link 81A is increased with respect to the movement amount of the movable joint 77A without changing the length of the link, The enlargement ratio can be improved.

  The control device 5 will be described with reference to FIG. The control device 5 is a means for controlling the entire microinjection device, and is composed of a computer such as a microcomputer or a personal computer, a program executed on the computer, an electronic circuit, and the like. In addition to the position determination unit 3, the control device 5 includes a responsible body determination unit 87 serving as a control unit for the transport unit 4 and an injection operation control unit 88.

  The assigned introducer determining unit 87 determines the position of each introducer by the position determination means 3, for example, based on the positional relationship of each introducer in the entire image, based on the setting rule or the like. It is a means for determining a target in charge of injection into which to-be-introduced body and a target position for injecting the injection needle 11 into which position of the determined to-be-introduced body. The person in charge of the introducer need not be defined as “person in charge”, and the target position where the tip of each injection needle 11 moves is determined, and as a result, the person in charge of the introducer is determined. good. Further, the target position may be determined so that the same introduced object is injected by a plurality of injection needles 11.

  The injection operation control unit 88 is a unit that controls each transport unit 4 such that the tip of the injection needle 11 moves to the target position determined by the assigned introducer determination unit 87. The injection operation control unit 88 includes a conveyance unit individual control unit 88 a that performs drive control of the moving mechanisms 14 to 16 for each axis for each conveyance unit 4. When the injection operation control unit 88 has injection drive means (not shown) for discharging the introduction substance to the injection needle 11, in addition to the above control, control means (not shown) of the injection drive means. Is provided. In addition, the injection operation control unit 88 includes a retraction mechanism control unit 89 that controls each transport unit retraction mechanism 39 and a container position adjustment control unit 79 that controls each axis of the container position adjustment unit 1. .

Next, the injection operation by the microinjection apparatus having the above configuration will be described.
First, the container mounting table 13 is moved horizontally by driving the container position adjusting unit 1, and the container 12 on the container mounting table 13 is positioned at a predetermined position below the imaging unit 2. Next, the imaging unit 2 captures a partially enlarged plan view image in the container 12 through the lens 2a. The image is processed by the image processing means 7, and the position determination means 3 of the control device 5 determines the position of the cell that is the introducer in the container 12 from the image.

Next, according to the position information of each cell obtained by the position determination means 3, the injected body determination unit 87 causes the cells to be injected by each injection needle 11 and the tip of the injection needle 11 as a cell. A target position to be inserted is determined. Along with this determination, each transport means 4 that supports the injection needle 11 is controlled by each transport means individual control section 88 a of the injection operation control section 88. In this case, the injection operation control unit 88 operates so that the injection needles 11 of the transfer means 4 do not interfere with each other.
Thereby, for example, the following injection operations by the injection needles 11 are performed in parallel. That is, as the injection operation, each injection needle 11 is positioned in the horizontal direction and the insertion direction, and the injection needle 11 is inserted into the cell determined by the movement of the injection needle 11 in the insertion direction. The filled introduction substance is introduced into the cells. This introduction may be performed by discharging the introduction substance from the injection needle 11 by driving, or may be introduced by permeation by osmotic pressure or the like. The entire series of injection operations by these injection needles 11 is controlled by the control device 5. The movement of the injection needle 11 in the insertion direction can be vibrated as a reciprocating movement in order to enable smooth insertion into the cell. Further, the frequency of the vibration can be changed according to the type of the cell, or depending on the puncture to the cell membrane or the nuclear membrane. The introduced substance is a gene regulatory factor such as DNA or protein.

  After the injection process to the cells in the image is completed, the container mounting table 13 is horizontally moved by a predetermined amount by the operation of the container position adjusting means 1, so that another position close to the previous image capturing position in the container 12 is obtained. The position determination unit 3 recognizes the cell position at 1 from the image obtained by the imaging unit 2, and the above-described injection operation is repeated.

  In particular, in this microinjection apparatus, the conveying means 4 has a three-degree-of-freedom moving mechanism (X-axis moving mechanism 14, Y-axis moving mechanism 15, Z-axis moving mechanism 16), and drives these moving mechanisms 14-16. Since the stacked piezoelectric elements (piezoelectric element stacks 19A to 19J) are used as the source, the transport means 4 can be reduced in size, and a plurality of transport means 4 are provided around the container 12 of the introduced body such as a petri dish. Can be arranged, and the effect can be further improved when performing the injection operation to a plurality of cells at the same time. The transport unit 4 has a moving mechanism with at least two degrees of freedom. If a stacked piezoelectric element is used as a driving source with at least one degree of freedom among the moving mechanisms with each degree of freedom, the transport unit 4 can be downsized. Become. Piezoelectric elements have a high conversion efficiency for converting electrical energy into mechanical energy, and can change the generated displacement relatively easily by changing the applied voltage, and have excellent controllability.

  Moreover, this microinjection apparatus can be set as the structure with high injection process efficiency only by adding the some conveyance means 4 corresponding to the some injection needle 11 in the prior art example shown in FIG.

  As described above, in order for the tip of the injection needle 11 to move within the predetermined visual field in the image obtained by the imaging means 2, the movement distance of the injection needle 11 by the transport means 4 is 1 mm or more. Is desirable. In this embodiment, two sets of stacked piezoelectric elements (piezoelectric element stacks 19E, 19F or 19I, 19J, Y axis in the X-axis moving mechanism 14) are used as driving sources for the moving mechanisms 14, 15, 16 in the transport means 4. Piezoelectric element laminates 19C, 19D or 19G, 19H in the moving mechanism 15 and piezoelectric element laminates 19A, 19B) in the Z-axis moving mechanism 16 are arranged in parallel along the lamination direction of the piezoelectric elements, and these are connected to the fastening member 48. , 58, 62, 78, and 85, it is possible to ensure a sufficient amount of movement for the injection needle 11 without requiring a large space.

  Further, in this embodiment, in the X-axis moving mechanism 14 and the Y-axis moving mechanism 15 in the transport unit 4, the piezoelectric element stack (piezoelectric element stacks 19E and 19F or 19I, 19J and Y-axis moves in the X-axis moving mechanism 14). The mechanism 15 has an expansion mechanism that expands the displacement of the piezoelectric element laminate 19C, 19D or 19G, 19H to a displacement in the direction (Y-axis direction) orthogonal to the displacement direction (X-axis direction). This is more effective in securing the amount of movement required for the injection needle 11.

  Furthermore, in this embodiment, in the X-axis moving mechanism 14 and the Y-axis moving mechanism 15 in the transport unit 4, the piezoelectric element stack (piezoelectric element stacks 19E, 19F or 19I, 19J, Y-axis shift in the X-axis moving mechanism 14) is used. In the mechanism 15, the displacement of the piezoelectric element laminate 19C, 19D or 19G, 19H) is expanded to a displacement in a direction orthogonal to the displacement direction, and the displacement expanded by the first expansion mechanism is Since it has the 2nd expansion mechanism which expands to the displacement of the displacement direction (X-axis direction) of a piezoelectric element laminated body, it becomes further effective in securing the movement amount required for the injection needle 11.

  In this embodiment, as shown in FIG. 4, the Z-axis moving mechanism 16 that moves the needle support member 18 that supports the injection needle 11 in the insertion direction (Z-axis direction) of the injection needle 11 serves as a drive source. Vibration driving means 54 is provided to repeatedly change the displacement of a part of the piezoelectric element laminates 19B1 in the piezoelectric element laminates 19A and 19B to vibrate the needle support member 18 in the insertion direction, and the piezoelectric element laminates 19A and 19B. Are used separately for vibration and positioning applications. For this reason, the injection needle 11 can be smoothly stabbed into a cell to be a derivative, and the positioning of the injection needle 11 can be driven at high speed.

  In the configuration example of the Z-axis moving mechanism 16 shown in FIG. 5, the vibration driving means 54 superimposes a positioning signal for positioning the needle support member 18 and a vibration drive signal for vibrating the needle support member 18 in the insertion direction. And since it applies to some piezoelectric element laminated bodies 19B1, some piezoelectric element laminated bodies 19B1 can be shared by vibration provision and positioning.

  Further, in this embodiment, since the part of the piezoelectric element laminate 19B1 contributing to the vibration is directly connected to the needle support member 18, the displacement due to the vibration generated in the piezoelectric element laminate 19B1 is injected. 11 can be efficiently transmitted.

  Further, in this embodiment, the vibration driving means 54 of the Z-axis moving mechanism 16 includes the frequency variable means 55 for switching the frequency of vibration, so that the cell that is the introducer into which the injection needle 11 is inserted is inserted. The frequency of the vibration can be switched according to the type, cell membrane, and nuclear membrane, and the injection needle 11 can be smoothly inserted accordingly.

  By the way, the cells to be introduced include adhering cells attached to a medium and the like and floating cells floating in the culture solution. In the case of adherent cells, the injection operation can be performed according to the procedure described above. FIG. 12 and FIG. 13 show an example of the configuration of the cell adsorption substrate unit used when the introducer is a floating cell. The cell adsorption substrate unit 66 is formed by closing the opening of a box 67 having an open upper surface with a cell adsorption substrate 68 having a plurality of fine holes 69, and a part of the box 67 includes a box 67. A suction port 70 is provided for sucking the culture solution from the inside. As the cell adsorption substrate 66, those having different diameters of the micropores 69 are prepared depending on the size of the cells to be injected. In the microinjection apparatus shown in FIG. 1, an example is shown in which a pump 72 is provided when the cell adsorption substrate unit 66 is used. By sucking the culture solution from the suction port 70 of the cell adsorption substrate unit 66 with this pump 72, the floating cells 71 can be fixed in the micropores 69 of the cell adsorption substrate 68. Means (not shown) for controlling the pump 72 is also provided in the control device 5.

  In addition, an injection method that does not use the above-described cell adsorption substrate unit 66 when the introducer is a floating cell will be described. By combining at least two or more transport means similar to the transport means 4 of the injection needle 11, one side supports the injection needle 11, the other holds the cell fixing tube, and the floating cells are sucked from the tube. The injection operation is performed with the injection needle 11. In this case, the cell can be moved to an arbitrary position in the container 12 by an operation on the cell fixing tube side. That is, it becomes possible to sort the cells by operating the conveying means 4 holding the cell fixing tube.

  According to this microinjection apparatus, when the introduced substances filled in the plurality of injection needles 11 are the same substance in all of the injection needles 11, the speed of the injection operation is compared with the number of injection needles 11 compared to the conventional one. Can be doubled. In addition, when the introduction substances filled in the plurality of injection needles 11 are different substances for each injection needle 11, a plurality of types of introduction substances are introduced into one cell by repeatedly injecting the cells in order. It becomes possible to inject. The injection operation control unit 88 shown in FIG. 1 may be controlled to sequentially operate the plurality of transport means 4 in this way.

DESCRIPTION OF SYMBOLS 1 ... Container position adjustment means 2 ... Imaging means 3 ... Position determination means 4 ... Conveyance means 11 ... Injection needle 12 ... Container 14 ... X-axis movement mechanism 15 ... Y-axis movement mechanism 16 ... Z-axis movement mechanism 19A-19J ... Piezoelectric element Laminate 26 ... movable piece (enlargement mechanism)
36 ... movable piece (first enlargement mechanism)
37 ... Movable frame (second enlargement mechanism)
48, 58, 62 ... fastening member 54 ... vibration drive means 75 ... link mechanism (enlargement mechanism)
76 ... fixed joints 77A and 77B ... movable joints 81A to 81C ... link 101 ... first link mechanism (first expansion mechanism)
102 ... Second link mechanism (second enlargement mechanism)
106 ... fixed joints 107A and 107B ... movable joints 108A and 108B ... links

Claims (19)

A microinjection device that introduces a substance to be introduced into a body to be introduced by inserting a minute injection needle filled with the substance to be introduced into the body,
Container position adjusting means for adjusting the position of the container in which the introduced body is accommodated, and a plurality of injection needles can be individually moved with respect to the introduced body in the container whose position is adjusted by the container position adjusting means. A plurality of conveying means to be moved to, an imaging means for taking an image of the inside of the container whose position is adjusted by the container position adjusting means through an enlargement lens, and an image of the body to be introduced from the image obtained by the imaging means. Position determining means for determining the position, and control means for causing the respective conveying means to move the injection needle in accordance with position information obtained by the position determining means, wherein the conveying means has at least two degrees of freedom. And having a moving mechanism for each degree of freedom, and among these moving mechanisms for each degree of freedom, a plurality of piezoelectric elements are stacked on the driving source of the moving mechanism for at least one degree of freedom. A microinjection apparatus using a piezoelectric element laminate that expands and contracts.   2. The moving mechanism using the piezoelectric element laminate in the transport unit as a drive source according to claim 1, wherein a plurality of piezoelectric element laminates are arranged in parallel, and the plurality of piezoelectric element laminates are expanded and contracted via a fastening member. Microinjection device connected in series in the direction.   3. The micro mechanism according to claim 1, wherein the moving mechanism using the piezoelectric element stack in the transport unit as a drive source includes an expansion mechanism that expands the expansion and contraction of the piezoelectric element stack to a displacement perpendicular to the expansion and contraction direction. Injection device.   4. The microinjection device according to claim 3, wherein the expansion mechanism is a link mechanism.   5. The link mechanism according to claim 4, wherein the link mechanism includes three links, one fixed joint that is fixed in position, and three movable joints that are movable in position, and each joint constitutes a rotatable node. A first joint whose rotation center is perpendicular to both the expansion direction and the direction orthogonal to the expansion direction of the piezoelectric element laminate, and a base end of which is connected to the fixed joint; The other end of the second link is connected to the middle of the second link whose base end is connected to the first movable joint via the second movable joint, and the third movable joint is connected to the tip of the second link. The third link is connected, and the tip of the third link is a movable part that is constrained to be movable only in the direction orthogonal to the expansion and contraction direction, and the fixed joint is on the fixed side of the piezoelectric element laminate. end The first movable joint is movable integrally with the expansion / contraction side end of the piezoelectric element laminate, and the movable portion at the tip of the third link is connected to the displacement expansion output portion of the link mechanism. Microinjection device.   5. The link mechanism according to claim 4, wherein the link mechanism includes two links, one fixed joint that is fixed in position, and two movable joints that are movable in position, and each joint constitutes a rotatable node. A joint, the rotation center of which is perpendicular to both the expansion direction and the direction orthogonal to the expansion direction of the piezoelectric element laminate, and a first end whose base end is connected to a fixed joint The other end of the link is connected to the middle of the second link whose base end is connected to the first movable joint via a second movable joint, and the fixed joint is an end on the fixed side of the piezoelectric element laminate. The first movable joint can be moved integrally with the expansion / contraction end of the piezoelectric element laminate, and the tip of the second link serves as a displacement expansion output portion of the link mechanism. Njekushon apparatus.   In Claim 1 or Claim 2, the moving mechanism that uses the piezoelectric element laminate in the transport unit as a drive source includes a first expansion mechanism that expands and contracts the piezoelectric element to a displacement in a direction perpendicular to the expansion and contraction direction; A microinjection apparatus comprising: a second expansion mechanism that expands the displacement expanded by the first expansion mechanism to a displacement in the expansion / contraction direction of the piezoelectric element.   8. The microinjection device according to claim 7, wherein the first and second enlargement mechanisms are link mechanisms. 9. The link mechanism constituting the first and second magnifying mechanisms according to claim 8, wherein each of the link mechanisms includes three links, one fixed joint for position fixing, and three movable joints for position movement. Each of the joints is a joint that constitutes a rotatable node, and the center of rotation is perpendicular to both the expansion / contraction direction of the piezoelectric element laminate and the direction orthogonal to the expansion / contraction direction. The other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via the second movable joint. The third link is connected to the tip of the second link via a third movable joint, and the tip of the third link is restrained so as to be movable only in the direction orthogonal to the expansion / contraction direction. age,
In the link mechanism constituting the first expansion mechanism, the fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided on the expansion / contraction side of the piezoelectric element laminate. Can move together with the end,
The link mechanism constituting the second enlargement mechanism is provided in a posture in which the direction of 90 ° is changed around the central axis parallel to the rotation center of each joint with respect to the link mechanism constituting the first enlargement mechanism, The fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided in the movable portion of the link mechanism constituting the first expansion mechanism;
The microinjection apparatus in which the movable portion of the third movable joint of the link mechanism constituting the second expansion mechanism is a displacement expansion output portion in the expansion / contraction direction of the piezoelectric element laminate. In Claim 8, each of the link mechanisms constituting the first and second expansion mechanisms includes two links, one fixed joint for position fixing, and two movable joints for position movement. Each of the joints is a joint that constitutes a rotatable node, and the center of rotation is perpendicular to both the expansion / contraction direction of the piezoelectric element laminate and the direction orthogonal to the expansion / contraction direction. Yes, the other end of the first link whose base end is connected to the fixed joint is connected to the middle of the second link whose base end is connected to the first movable joint via the second movable joint,
In the link mechanism constituting the first expansion mechanism, the fixed joint is fixed in position with respect to the fixed side end of the piezoelectric element laminate, and the first movable joint is provided on the expansion / contraction side of the piezoelectric element laminate. Can move together with the end,
The link mechanism constituting the second enlargement mechanism is provided in a posture in which the direction of 90 ° is changed around the central axis parallel to the rotation center of each joint with respect to the link mechanism constituting the first enlargement mechanism, The fixed joint is fixed in position relative to the fixed-side end of the piezoelectric element laminate, and the first movable joint is provided at the tip of the second link in the link mechanism constituting the first expansion mechanism;
A microinjection device in which a tip of a second link of a link mechanism constituting a second enlarging mechanism serves as a displacement enlarging output unit in the expansion / contraction direction of the piezoelectric element laminate. In Claim 4, the said link mechanism has a crank slider mechanism which consists of one fixed joint, two movable joints, and two links,
This crank-slider mechanism converts the displacement in the extension direction of the piezoelectric element into an arbitrary direction on the circumference of the fixed joint via the two movable joints and the two links, thereby further expanding the displacement. Microinjection device.   12. The microinjection device according to claim 11, wherein the crank / slider mechanism is an offset crank mechanism.   3. The moving mechanism according to claim 1 or 2, wherein the moving mechanism that moves the needle support member that supports the injection needle in the insertion direction of the injection needle is a movement mechanism that uses the piezoelectric element laminate as a drive source. A microinjection device having vibration drive means for applying vibration in the insertion direction to the needle support member by driving a part of the piezoelectric elements constituting the vibration.   14. The vibration drive unit according to claim 13, wherein the vibration drive means superimposes a positioning signal for positioning the needle support member and a vibration drive signal for vibrating the needle support member in the insertion direction, and applies them to the partial piezoelectric elements. Microinjection device.   15. The microinjection device according to claim 13, wherein the piezoelectric element that is vibrated by the vibration driving unit is a piezoelectric element that is located at a connection side end with respect to the needle support member in the piezoelectric element laminate.   16. The microinjection device according to any one of claims 13 to 15, wherein the vibration driving means includes frequency variable means for switching a frequency of vibration.   17. The piezoelectric element according to claim 1, wherein a part of the piezoelectric elements that are driven by vibration includes a single piezoelectric element laminate, and is linearly connected to the other piezoelectric element laminate. Microinjection device.   18. The connection body according to claim 17, wherein one piezoelectric element multilayer body, which is a part of the piezoelectric elements driven by vibration, and another piezoelectric element multilayer body are linearly coupled, and further parallel to another piezoelectric element multilayer body. A micro-injection apparatus in which the coupling body and the other piezoelectric element laminated body that are arranged in parallel with each other and the other piezoelectric element laminated body are connected in series in the expansion / contraction direction via a fastening member.   The microinjection device according to any one of claims 1 to 18, wherein the introducer is a cell and the introduced substance is a gene regulatory factor.

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