WO2012008407A1 - Microinjection device - Google Patents

Abstract

Disclosed is a microinjection device that can improve processing efficiency and that can efficiently and reliably introduce an introduced substance to a transductant such as cells by means of a plurality of injection needles. The device is provided with: a plurality of conveyance means (4) that can move the plurality of injection needles (11) each individually; an imaging means (2) that captures an image of the inside of a vessel (12) through a microscope (3) at a microscope field of view (V); a position determination means (81) that determines the position of each transductant (T) in the microscope field of view (V) from the image; and an assigned range computation unit (82) that decides the assigned range (E) within the microscope field (V) to which each injection needle (11) is assigned, in a manner so that the load on the conveyance means (4) of each injection needle (11) is equalized according to prescribed rules (R).

Description

Microinjection device Related applications

This application claims the priority of Japanese Patent Application No. 2010-161319 filed on July 16, 2010, which is incorporated herein by reference in its entirety.

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 under a microscope.

Typical methods for introducing substances into cells include electroporation (electrical), lipofection (chemical), vector method (biological), microinjection method (mechanical), and laser injection (optical). . In the electrical method, the cell membrane is broken by a large current, so that the cell is greatly damaged. The chemical method is limited in the genes that can be introduced, and the introduction efficiency is low. Biological systems have limitations such as the types of genes that can be introduced and safety issues. The microinjection method has an advantage that a substance can be reliably injected into a cell by controlling the injection position with high accuracy. Patent Document 1 describes a method of performing microinjection using a capillary (microneedle).

Although the above-mentioned method can be cited as a method for introducing a gene into a cell, there is still no technology having both certainty and efficiency. The microinjection method, in which a gene is injected into each cell, is the most reliable method. However, the operation requires skill and time, so that the throughput is low. As a technique for increasing the throughput of the microinjection method, a microcapillary array in which a large number of injection needles are regularly arranged as described in Patent Document 2 has been developed, and a microchamber array having a chamber at a position corresponding to this is used. There is an example of trying batch injection.

FIG. 31 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

Since the shape, size, and elastic force of the cells are different, the microcapillary array system such as Patent Document 2 that collectively controls the position of the injection needle with respect to the cells does not pierce many cells, or many As a result, the cells are destroyed. In addition, since the method using a microcapillary array requires cells to be arranged at the same position as the microcapillary array, it can only handle floating cells that can move in the culture medium. Cannot be used.

In order to increase the throughput of the microinjection method, the injection speed of the injected substance is improved and the positioning speed of the manipulator (injection needle conveying means) is improved. There is one that has realized the injection process of one cell in less than 1 second. However, in medical use requirements, it is required that the type of introduced substance is not limited, that introduction efficiency is high, and that a large amount of substance-introduced cells can be supplied. It cannot be satisfied at present.

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 the manipulator equipped with the injection needles is large at present. In addition, in current microinjection devices, a petri dish stage containing cells is operated for cell positioning, and a manipulator equipped with an injection needle operates only in the direction in which the injection needle is inserted into the cell. I couldn't do it at the same time.

For example, in the current microinjection apparatus as described above with reference to FIG. 31, the container position adjusting means 94 that supports the petri dish 90 containing the cells is moved in order to position the cells, and the manipulator equipped with the injection needle 95 is moved to the cells. Since the injection needle 95 is moved only in the insertion direction, it was impossible to position a plurality of cells and insert the injection needle into each cell simultaneously. In addition, when the injection needle 95 moves in two degrees of freedom in the orthogonal X-axis direction and Y-axis direction in the conveying means of the injection needle 95, for example, the Y-axis movement mechanism is arranged to be stacked on the X-axis movement mechanism. It was. Therefore, since the X movement mechanism drives both the injection needle and the Y axis movement mechanism, high-speed movement is difficult.

The applicant previously proposed a reduction in the size of the manipulator, which was solved by using a piezoelectric element laminate as a driving source or using a mechanism for expanding the operation of the piezoelectric element laminate. (For example, Japanese Patent Application No. 2010-077677). In this proposed example, not only the direction of puncturing the injection needle, but also movement in the horizontal plane and movement in the puncturing direction are possible, and parallel injection processing to a plurality of cells is possible. In addition, there are other possible miniaturizations of manipulators, and the possibility of implementing a plurality of manipulators is increasing.

However, it has not been solved which injection target is to be injected using each of the manipulators arranged in plural. How to properly use a plurality of manipulators greatly affects the total processing time when injection processing is performed on all the introduced objects in the petri dish.

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 apparatus according to the present invention is a microinjection apparatus that performs an injection process for introducing an introduction substance into the introduction target T by inserting the injection needle 11 filled therein with the introduction substance into the introduction target T. The container position adjusting means 1 for adjusting the position of the container 12 containing the introduction target T, and a plurality of injections with respect to the introduction target T inside the container 12 adjusted by the container position adjusting means 1 A plurality of conveying means 4 for individually moving the needles 11 so as to be movable in at least two-dimensional directions, and images inside the container 12 adjusted by the container position adjusting means 1 are transmitted through the microscope 3 to the microscope visual field range V. Position determination for determining the position of each introducer T in the microscope visual field range V from the image obtained by the imaging means 2 and the image obtained by the imaging means 2 Each injection so as to equalize the load on the conveying means 4 of each injection needle 11 from the stage 8 and information on the position of each introduction target T determined by the position determination means 8 in accordance with a predetermined rule R. Position determination means 9 for determining the range E within the microscope visual field V that the needle 11 is in charge of, and for each introducer T in the range E determined by the charge range calculation means 9 And injection needle movement control means 88 for causing the respective conveying means 4 to move the injection needle 11 from the information on the position of each introduction target T determined by the means 8.

According to this configuration, since the plurality of transfer means 4 for individually moving the plurality of injection needles 11 so as to be movable in at least two-dimensional directions are provided, the plurality of injection needles 11 are operated in parallel to perform the injection process. be able to. The container position adjusting means 1 for adjusting the position of the container 12 containing the introduction target T is provided, and an image inside the container 12 adjusted by the container position adjusting means 1 is taken in the microscope visual field range V through the microscope 3. Imaging means 2 for performing the operation and position determining means 8 for determining the position of each introduced object T in the microscope visual field range V from the image obtained by the imaging means 2 are provided. By performing the injection process with the injection needle 11, it is possible to perform the injection process for all the introduced objects T in the container 12.

In this case, assigned range calculation means 9 for determining the range E in the microscope visual field V that each injection needle 11 is in charge of is provided, and this means 9 is provided in accordance with a predetermined rule R of the conveying means 4 of each injection needle 11. Since the assigned range is determined so that the load is equalized, the injection process can be performed on the entire introduced target T in the microscope visual field V without the load being unevenly applied to some of the injection needles 11. . For this reason, the time difference for completing the injection process for all the introduction bodies T in charge of each injection needle 11 to be operated in parallel within the microscope visual field V is reduced. Therefore, the introduction of the introduced substance into the introduction target T can be performed efficiently and reliably with the plurality of injection needles 11, and the processing efficiency can be improved.

The “load” is a physical quantity that is a factor that affects the operation time and the number of operations of the conveying means of the injection needle 11, and is a physical quantity that is calculated according to an arbitrarily defined rule. Or the number of times the injection needle 11 moves, or a value that takes into account both the distance and the number of times.

In this invention, the assigned range calculation means 9 may divide the region of the microscope visual field range V so that the area of the range E handled by each injection needle 11 is substantially equal. If the area of the range E in charge is substantially equal, the load of the conveying means 4 of each injection needle 11 is equalized, and the time of the injection process by each injection needle 11 is equalized. In addition, since the area of the range E in charge is divided almost evenly, the division calculation process is easy.

In the present invention, the assigned range calculation means 9 may divide the region of the microscope visual field range V so that the number of introduced bodies T handled by each injection needle 11 is substantially equal. If the number of introduced objects T in charge is equal, the number of operations for the injection process of the injection needles 11 becomes equal, and therefore the time of the injection process by each injection needle 11 is equalized.

In the present invention, the order of the injection process of the introduced body 11 is set so that the total movement distance of the injection needle 11 in the operation of the injection process to each introduced body T in charge is minimized for each injection needle 11. Insertion order determining means 10 for determining may be provided. In the operation of the injection process, the injection needle 11 is adjusted and moved so that the injection needle 11 comes to a position facing the introduction target T while the injection needle 11 is in the standby position in the insertion direction, and the injection needle 11 is inserted into the introduction target. The operation in the insertion direction and the operation of returning the introduction target T to the standby position are repeated. When performing injection processing on a plurality of introducers T within the assigned range E, the order of processing is less affected by the time of operation in the insertion direction, but the distance of the adjustment movement differs greatly. The sum of travel time changes greatly. Therefore, by determining the order so that the total movement distance of the injection needle 11 is the shortest, the total time for performing the injection processing of all the introduced objects in the assigned range E can be shortened.

When the insertion order determination means 10 is provided, the assigned range calculation means 9 may be configured as follows. For example, the assigned range calculation means 9 divides the region of the microscope visual field range V so that the total movement distances of the individual injection needles 11 are substantially equal. When the insertion order determining means 10 is provided, the total movement distance of the individual injection needles 11 is determined. By equalizing the total moving distances determined in this way, the time of the injection process that each injection needle 11 takes charge is equalized, and the processing efficiency can be improved.

In the case where the insertion order determining means 10 is provided, the assigned range calculating means 9 divides the region of the microscope visual field range V so that the product of the total moving distance and the processing number of the individual injection needles 11 is substantially equal. It may be what you do. The time required for the movement of the injection needle 11 for the injection process includes a time for adjusting and moving the injection needle 11 so as to come to a position facing the introduction target T and a time for reciprocating the injection needle 11 in the insertion direction. is there. The time for reciprocating the injection needle 11 in the insertion direction depends on the number of processing. The time for the adjustment movement depends on the total movement distance. Therefore, by dividing the region of the microscope field of view so that the product of the total moving distance and the number of treatments is almost equal, the injection processing time assigned to each injection needle is further equalized and the processing efficiency is further improved. Is possible. Further, as described above, the processing time is determined from both the processing number and the total moving distance, but by using the product of both, an appropriate area division reflecting the processing number and the total moving distance can be easily performed.

In the case where the insertion order determining means 10 is provided, the assigned range calculating means 9 is used for filling the introduction substance while the injection needle 11 is being moved and while the injection needle 11 is being inserted into the introducer T. The region of the microscope field-of-view range V may be divided so that the processing time, which is the sum of the injection time and the movement time, is substantially equal using the injection time to be stopped. By considering the injection time when the injection needles 11 are stopped due to the filling of the introduced substance in addition to the movement time, the time of the injection process in which each injection needle 11 is in charge is further equalized and the processing efficiency is improved. Further improvements are possible.

In the present invention, the assigned range calculation means 9 has a priority condition that no interference occurs between the injection needles 11 when the injection needles 11 are operated in parallel. Under this priority condition, the microscope visual field range V The area may be divided. When the injection needles 11 interfere with each other when the injection needles 11 are operated in parallel, the injection needles 11 cannot be operated in parallel. Therefore, it is a priority condition that interference does not occur, and within the range where this priority condition is satisfied, other conditions, for example, the area in charge is almost equal, or the number of introduced subjects in charge is almost equal. It is practical to divide the region of the range.

In the present invention, interference avoiding means 79 may be provided for determining the order of the injection processing of each injection needle 11 to the introduction target T so that the injection needles 11 do not interfere with each other. Thereby, avoidance of interference between the injection needles 11 is ensured.

A visual field range adjusting means 86 for adjusting the microscope visual field range V may be provided according to the size of the introduction target T. The microscope visual field range V is adjusted by changing the magnification of the objective lens of the microscope 3, for example. Depending on the size of the body to be introduced T, the accuracy requirement of the insertion position of the injection needle 11 into the body to be introduced T varies. Therefore, when the introducer T is large, the microscope visual field range V is widened to reduce the number of times of changing the container position, and when the introducer T is small, the microscope visual field range V is narrowed to improve accuracy and necessary for introduction. The efficiency of the injection process can be improved without causing a significant decrease in accuracy.

The container position adjusting means 1 and the container position adjusting means 1 sequentially move the position of the container 12 according to the microscope visual field range V so that the injection processing of the injection needle 11 is performed in the entire region of the container 12. An in-container full range processing means 100 for controlling the conveying means 4 of each injection needle 11 may be provided. Thereby, the introduction substance can be introduced into all the introduction bodies T in the container 12.

The in-container full range processing means 100 performs the injection processing on all the objects to be introduced T in one microscope visual field range V and then moves the container 12 by the container position adjusting means 1 according to the microscope visual field range V. You may make it move sequentially and to perform the injection process with respect to the to-be-introduced object T in the container 12 in the position after each movement. In this case, only one microscope field-of-view range V needs to be stored for the position of the introducer T, and the storage capacity can be reduced.

In the case where the in-container all-range processing means 100 is provided, the in-container all-range processing means 100 includes an insertion order determining means 10 that determines the order of the injection processing of each injection needle 11 according to a predetermined rule. The position of the container 12 is sequentially moved by the container position adjusting means 1 in accordance with the microscope visual field range V, and the position of the introduction target T up to each microscope visual field range V of a plurality of movement destinations is set before the injection process. The position determination means 8 may make the determination, and the assigned range calculation means 9 may determine the range E that each injection needle 11 is responsible for, and the insertion order determination means 10 may determine the order of the injection processing. . By acquiring the information on the position of the target T to be moved ahead in this way, the movement destination of the injection needle can be calculated in advance during the injection process of the microscope visual field range V. The processing time can be shortened.

When the in-container full-range processing means 100 is provided, the in-container full-range processing means 100 is provided with an insertion order determining means 10 that determines the injection processing order of the injection needles 11 according to a predetermined rule. The position of the container 12 is sequentially moved by the container position adjusting means 1 according to the visual field range V, and the positions of all the introduced objects T in the container 12 are determined by the position determining means 8 before the injection process. In addition, the range calculation means 9 may determine the range E that each injection needle 11 is responsible for, and the insertion order determination means 10 may determine the order of the injection processing. By determining the positions of all the introduced objects T in the container 12 by the position determining means 8 before the injection process, it is possible to perform more efficient region division, determination of the injection process order, and the like.

The in-container full range processing means 100 sequentially moves the position of the container 12 by the container position adjusting means 1 in accordance with the microscope visual field range V, and determines the position of the introducer T in the current microscope visual field range V as a position determination means. At the same time as the determination by step 8, the injection process to each introduced object T in the microscope visual field range V a plurality of times before may be performed by the movement of the transfer device 4 by the injection needle movement control means 88. In the case of this configuration, the position of the introducer T can be calculated during the injection process, and the entire injection process can be performed more efficiently.

The in-container full range processing means 100 may be moved so as to have a portion Va overlapping the microscope visual field range V when the container position adjusting means 1 sequentially moves the 12 positions of the container. By providing the overlapping portion Va, it is possible to prevent the unprocessed material T from being generated in the vicinity of the boundary of the microscope visual field range V.

When the container 12 is moved so as to have an overlapping portion Va in the microscope field-of-view range V, the entire range processing means 100 in the container may be configured so that the overlapping portions Va have the same area. By making the area of the overlapping portion Va the same, the process of dividing the range in the container into a large number of microscope visual field ranges V is facilitated.

When moving so as to have a portion Va overlapping the microscope visual field range V, the in-container all-range processing means 100 determines the rule when the container position adjusting means 1 sequentially moves the position of the container 12. Thus, it is also possible to determine a to-be-introduced object To in the microscope visual field range V and move it so that the regions including the to-be-introduced object To overlap. In the case of this configuration, the introduced object To at the overlapping position is the reference, and the position of the introduced object To becomes clear.

In the present invention, the transport means includes a movable body installed so as to be movable forward and backward, and an advance / retreat drive means for moving the movable body, and the advance / retreat drive means includes a plurality of piezoelectric elements stacked as a drive source. It is also possible to have a piezoelectric element laminate that expands and contracts in the lamination direction. When a laminated piezoelectric element is used as a driving source, the conveying means can be further reduced in size, a large number of conveying means can be arranged in a limited space around the container, and a large number of injection needles are used. Thus, the efficiency of the injection process can be improved.

When the piezoelectric element laminate is used as a drive source, the advancing / retreating drive unit using the piezoelectric element laminate as a drive source in the transport unit arranges the piezoelectric element laminate in parallel, and the plurality of piezoelectric element laminates May be connected in series in the expansion and contraction direction via a coupling member. 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 this configuration, the advancing / retracting drive means using the piezoelectric element laminate in the transport means as a drive source has an expansion mechanism that expands and contracts the piezoelectric element laminate to a displacement in a direction perpendicular to the expansion / contraction direction. Also good. The enlargement mechanism is, for example, a link mechanism. Having an enlargement mechanism is more effective in securing the amount of movement required for the injection needle. According to the link mechanism, it is possible to provide an expansion mechanism with a simple configuration.

In the case where the piezoelectric element laminate is used as a drive source, the advancing / retreating drive unit using the piezoelectric element laminate as the drive source in the transport unit is configured to extend and contract the piezoelectric element laminate in a direction parallel to the extension / contraction direction. It is good also as what has an expansion mechanism which expands to this displacement. The enlargement mechanism is, for example, a link mechanism. Enlarging the displacement in a direction parallel to the expansion / contraction direction is also more effective in securing the amount of movement necessary for the injection needle.

In any of the cases where the piezoelectric element laminate has an expansion mechanism that expands the expansion / contraction of the piezoelectric element laminate to a displacement in a direction perpendicular to the expansion / contraction direction and the expansion mechanism that expands the displacement in a direction parallel to the expansion / contraction direction The following configuration can be adopted. That is, the link mechanism has a crank / slider mechanism including one fixed joint, two movable joints, and two links, and the crank / slider mechanism is configured to change the displacement of the piezoelectric element in the extension direction. It may be possible to change the displacement in an arbitrary direction on the circumference of the fixed joint and further expand the displacement via two movable joints and two links. When the link mechanism has such a configuration, the number of parts can be reduced and the size can be reduced.

In the present invention, the conveying means has two degrees of freedom in which at least the first moving body and the second moving body can freely advance and retreat in directions orthogonal to each other, and the injection needle is attached to the second moving body. The first moving body is supported on the base so as to be movable in a linear direction via the first guide, and is movable on the first moving body via the second guide. A body is installed so as to be able to advance and retract in a direction orthogonal to the linear direction, and the first advancing / retreating driving means and the second advancing / retreating driving means for advancing and retracting the first moving body and the second moving body, respectively, The output member of the second advancing / retreating drive unit may be connected to or freely contact with a direction perpendicular to a direction in which the second moving body can advance / retreat. .

In the case of this configuration, the conveying means separates the moving body and the advancing / retreating driving means, and the advancing / retreating driving means for each degree of freedom are installed on the base, so that the conveying means can be reduced in size and the mass of the moving object is reduced. As a result, the conveying means can be driven at high speed. Although the advancing / retreating driving means for each degree of freedom are all installed on the base, the second advancing / retreating driving means of the second moving body installed on the base via the first moving body is provided. Since the output member is freely connected to or in contact with the direction orthogonal to the direction in which the advance / retreat movement is possible, the advance / retreat of the second advancing / retreating drive unit is performed regardless of the position of the first moving body. The drive can be transmitted to the second moving body. Further, since the conveying means can be reduced in size, it becomes easy to arrange a plurality of conveying means around a container such as a petri dish containing cells or other objects to be introduced. Therefore, the tip of the injection needle can be simultaneously positioned with respect to a plurality of introduced bodies, and injection processing to a large number of introduced bodies can be executed at a time, and high throughput can be achieved.

In the present invention, the introduced substance may be a cell, and the introduced substance may be a gene regulatory factor such as 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. .

Any combination of at least two configurations disclosed in the claims and / or the specification and / or drawings is included in the present invention. In particular, any combination of two or more of each claim in the claims is included in the present invention.

The present invention will be more clearly understood from the following description of preferred embodiments with reference to the accompanying drawings. However, the embodiments and drawings are for illustration and description only and should not be used to define the scope of the present invention. The scope of the invention is defined by the appended claims. In the accompanying drawings, the same reference numerals in a plurality of drawings indicate the same or corresponding parts.
(A) is explanatory drawing of the conceptual structure of the microinjection apparatus concerning one Embodiment of this invention, (B) is explanatory drawing of the division | segmentation by the microscope visual field range in the container, (C) is the charge range of the microscope visual field range. It is explanatory drawing of the division | segmentation into. It is a top view which shows the arrangement structure of the conveyance means in the microinjection apparatus. It is explanatory drawing which shows an example of the division | segmentation form of a microscope visual field range. It is explanatory drawing which shows the other example of the division | segmentation form of a microscope visual field range. It is explanatory drawing which shows the further another example of the division | segmentation form of a microscope visual field range. It is explanatory drawing which shows the further another example of the division | segmentation form of a microscope visual field range. (A) is explanatory drawing of the operation | movement which judges the position of all the to-be-introduced bodies in a container collectively, (B) is the operation | movement which judges the position of a to-be-introduced body collectively for every several divided | segmented range in a container. It is explanatory drawing. (A), (B) is explanatory drawing of the form which gives the overlap part of a microscope visual field range, respectively. It is a top view which shows the other arrangement structure of the conveyance means in the same microinjection apparatus. It is a side view of the conveyance means in the microinjection apparatus. (A) is a partially omitted plan view showing an X-axis moving mechanism and a Y-axis moving mechanism of the conveying means in the microinjection device, and (B) is a front view of the same. (A) is a top view which shows the X-axis movement mechanism and Y-axis movement mechanism of the conveyance means, (B) is the front view similarly. It is sectional drawing which shows an example of the advance / retreat drive means of the conveyance means. It is a top view which shows an example of the container position adjustment means in the microinjection apparatus. It is an enlarged plan view which shows an example of the contact part of the output member and moving body in the advance / retreat drive means of the conveyance means. It is an enlarged plan view which shows the other example of the contact part of the output member and moving body in the advance / retreat drive means of the conveyance means. (A) is a top view which shows the other example of the X-axis movement mechanism and Y-axis movement mechanism of the conveyance means in the microinjection apparatus, (B) is the front view similarly. (A) is a top view which shows the further another example of the X-axis movement mechanism of the conveyance means in the microinjection apparatus, and a Y-axis movement mechanism, (B) is the front view similarly. (A) is a top view which shows the further another example of the X-axis movement mechanism of the conveyance means in the microinjection apparatus, and a Y-axis movement mechanism, (B) is the front view similarly. (A) is a top view which shows the further another example of the X-axis movement mechanism of the conveyance means in the microinjection apparatus, and a Y-axis movement mechanism, (B) is the front view similarly. 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 of the other example shown combining the advance / retreat drive means in the conveyance means, and the block diagram of the conceptual structure of the control system. It is a figure which shows the further another example which combined the block diagram of the conceptual structure of the advancing / retreating drive means and its control system in the conveyance means. It is a figure which shows the further another example which combined the block diagram of the conceptual structure of the advancing / retreating drive means and its control system in the conveyance means. (A) is a plan view showing one configuration example of a link mechanism in the reciprocating drive means, (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 mechanism. It is a top view which shows the further another structural example. It is a figure which shows the further another example which combined the block diagram of the conceptual structure of the advancing / retreating drive means and its control system in the conveyance means. It is a top view which shows the further another structural example of the link mechanism in the advance / retreat drive means of the conveyance means. (A) is a diagram showing still another example in which the advance / retreat drive means (Y-axis movement mechanism) in the transport means and a block diagram of the conceptual configuration of the control system are combined, and (B) is the advance / retreat drive means in the transport means. It is a figure which shows the further another example which combined the block diagram of the conceptual structure of (X-axis movement mechanism) and its control system. It is a top view which shows one structural example of the link mechanism of advancing / retreating drive means. It is a top view which shows another structural example of the link mechanism. It is a conceptual diagram of a prior art example.

A first embodiment of the present invention will be described with reference to FIGS. 1A to 1C to FIG. 1A to 1C are conceptual diagrams of this microinjection apparatus. This microinjection apparatus is an apparatus for introducing an introduction substance into the introduction target T by inserting a minute injection needle 11 filled with the introduction substance into the introduction target T. The introducer T 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 each of these transfer units 4 between an injection preparation position and a retracted position. Are provided with a transporting means retracting mechanism 39 that moves forward and backward, an imaging means 2, a microscope 3, and a control device 5 that controls the operation of the entire apparatus.

The container position adjusting means 1 moves a container mounting table 13 on which a container 12 such as a petri dish that accommodates an introduction body is held in a horizontal state in two horizontal orthogonal directions (X-axis and Y-axis directions). 12 is a means for adjusting the position of the XY stage device 6. Note that the X axis, Y axis, and Z axis in this specification represent the respective axes of rectangular coordinates defined as a common coordinate system in the entire microinjection apparatus. May be represented as an axis for each degree of freedom.

The imaging means 2 is installed together with the microscope 3 at a fixed position above the container mounting table 13 so as to overlook the container 12. The imaging unit 2 is a camera or the like that captures an image such as a planar view image obtained by locally expanding the inside of the container 12 through the microscope 3, and performs imaging for each field of view range V (FIG. 1B) of the microscope 3. Do. The microscope visual field range V is set to the microscope visual field range V by moving the container 12 by the container position adjusting means 1. For example, each range obtained by dividing the inside of the container 12 as shown in FIG. The captured image is processed by the image processing means 7. The microscope 3 has a drive-type magnification adjusting means (not shown) that can adjust the field of view range by adjusting the magnification of an objective lens (not shown) by an input signal given from the outside.

The conveying means 4 is a means composed of an XYZ stage device or the like for moving each injection needle 11 and a plurality of conveying means 4 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 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, two) 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. As shown in FIG. 9, three conveying means 4 may be arranged such that three on the left and right are arranged at equal angular intervals, or more.

1A, the conveying means 4 is a mechanism having three degrees of freedom, and can move the injection needle 11 in a three-dimensional direction. Specifically, the transport unit 4 includes a mechanism that moves the injection needle 11 in two horizontal orthogonal directions and a mechanism that moves the injection needle 11 back and forth in a linear direction inclined with respect to the horizontal plane. A more specific configuration of the transport unit 4 will be described in detail later. In addition, as long as the conveyance means 4 can move in at least two degrees of freedom, that is, in the direction of two orthogonal axes, a plurality of conveying means 4 can be installed and subjected to the injection processing in parallel.

The control device 5 is constituted by a computer and a program executed by the computer. By these, the position determination means 8, the assigned range calculation means 9, the insertion order determination means 10, the target position determination means 80, the in-container full range processing command means 100, visual field range adjusting means 86, container movement interval determining means 87, injection needle movement control means 88, and container movement control means 89 are configured. Further, interference avoiding means 79 may be provided. In addition to this, the control device 5 includes input means by manual operation such as a keyboard and a mouse, input means for inputting from a communication line or a recording medium, and display means such as a liquid crystal display device capable of displaying an image (both shown (Not shown) is provided or connected.

The position determination means 8 is a means for determining the position of each introduced object T in the visual field range V of the microscope 3 from the image obtained by the imaging means 2. The image obtained by the imaging unit 2 is processed into a binary image or the like by the image processing unit 7, and the position determination unit 8 sets the black spot position of the binary image as the position of the introduction target T, for example. The position of the introducer T is determined as a coordinate position in the orthogonal biaxial direction within the microscope visual field range. From the relationship between the microscope visual field range V and the position of the container 12, the coordinate position of the introducer T with respect to the container 12 and the coordinate position of the introducer T with respect to the reference position of the container position adjusting means 1 are obtained. The position of each introduced object T determined by the position determination unit 8 is stored in the position storage unit 8 a in the position determination unit 8.

The assigned range calculation means 9 is a means for determining the range E (FIG. 1C) that each injection needle 11 takes charge in the microscope visual field V, and is determined by the position determination means 8 and stored in the position storage unit 8a. From the information on the position of each introduced object T, the assigned range E is determined according to a predetermined rule R. This rule R is to determine the assigned range E so that the load on the conveying means 4 of each injection needle 11 is equalized.

The determination of the assigned range E by the assigned range calculation means 9 is performed as shown in FIG. 3, FIG. 4, FIG. 5, or FIG. In any example, the visual field range V is injected by the radial dividing lines L around the radiation center O defined in the visual field range V so that the respective divided ranges are located close to the injection needles 11. It is assumed that the division range is divided into the number of needles 11 and the division range at the close position is set as the assigned range E of the injection needle 11. The radiation center O may be the center of the visual field range V, that is, the centroid, or may be an arbitrarily determined point. Under this assumption, it is determined as shown in each figure.

FIG. 3 is an example in which the region of the microscope visual field range V is divided into a plurality of assigned ranges E by a dividing line L around the radiation center O so that the areas handled by the injection needles 11 are almost equal. The radiation center O is the center of the visual field range V, but may be a position deviated from the center of the visual field range V.

In FIG. 4, the region of the microscope visual field range V is divided into a plurality of assigned ranges E by a dividing line L around the radiation center O so that the numbers of the introduction bodies T handled by each injection needle 11 are substantially equal. It is an example. The radiation center O is the center of the visual field range V in the above example, but may be a position deviated from the center of the visual field range V.

In FIG. 5, the region of the microscope visual field range V is divided into a plurality of assigned ranges E by a dividing line L around the radiation center O so that the total movement distances of the individual injection needles 11 by the conveying means 4 are substantially equal. It is an example. In this example as well, the radiation center O is the center of the visual field range V, but may be a position shifted from the center of the visual field range V. In addition, since the total movement distance changes if the order of performing the injection process on each introducer T within the assigned range E is different, the total movement when the order of the injection processes that minimizes the total movement distance is taken. Make the distances approximately equal. The order of the injection processing in which the total movement distance is the shortest is calculated by the insertion order determination means 10 (FIG. 1A). In this case, for example, it is possible to calculate the assigned range E in which the total moving distance is almost equal by determining the assigned range E variously and comparing the total moving distance in each case.

In addition to the above examples, the assigned range calculation means 9 can employ various methods for determining the assigned range E, which will be described later in the explanation section of the operation of the microinjection apparatus.

The insertion order determining means 10 introduces the introduction target in the assigned range E so that the total movement distance of the injection needle 11 in the operation of the injection process to each introduced target T for each injection needle 11 becomes the shortest. It is a means for determining the order of the injection processing of the body T.

The target position determining unit 80 is a unit that determines a target coordinate position into which the injection needle 11 is inserted with respect to each of the introduced objects T whose order has been determined by the insertion order determining unit 10. This coordinate position is determined from the coordinate position of the introducer T within the microscope visual field range V and the coordinate position of the microscope visual field range V with respect to the container position. The container position is a position moved by the container position adjusting means 1.

The injection needle movement control means 88 is means for controlling the transport means 4 so that the injection needle 11 moves to the target coordinate position determined by the target position determination means 80. The injection needle movement control means 88 is a general term for the individual injection needle control unit 88 a provided for each injection needle 11. These injection needle individual control units 88a are collectively referred to as injection needle movement control means 88. The injection needle individual control unit 88a controls each drive source in the X-axis direction, the Y-axis direction, and the Z-axis direction in the individual transport means 4.

The container movement control means 89 is a means for controlling the driving source in each axial direction (X direction, Y direction) which is the horizontal direction of the container position adjusting means 1 so as to be the coordinate position of the input container position in the XY direction. It is.

The in-container full range processing command unit 100 sequentially moves the position of the container 12 by the container position adjusting unit 1 according to the microscope visual field range V, and performs the injection processing of the injection needle 11 in the entire region in the container 12. It is a means to control the container position adjusting means 1 and the conveying means 4 of each injection needle 11 so that This control is performed via the injection needle movement control means 88 and the container movement control means 89 by giving a command to the injection needle movement control means 88 and the container movement control means 89. For example, as shown in FIG. 1B, the entire range of the container 12 is divided into regions in which the microscope visual field range V is arranged in a matrix in the vertical and horizontal directions, and sequentially positioned in each of the divided microscope visual field ranges V. The container 12 is moved and positioned by the container position adjusting means 1, and the conveying means 4 of each injection needle 11 is driven with respect to the microscope visual field range V at the stop position of each container 12. In addition, when arranging the microscope visual field range V in the range of the container 12, an overlapping range may occur as described later.

In this example, the in-container all-range processing command unit 100 moves the position of the container 12 every time the injection processing in one microscope field-of-view range V is completed, and introduces in the next one microscope field-of-view range V. The operation of determining the position of the body T and performing the injection process is repeated. In addition, as in the example described later, the determination of the position of the introduction target T in the entire range in the container 12 or in a plurality of microscope field-of-view ranges V may be performed collectively.

The visual field range adjusting means 86 is a means for adjusting the microscope visual field range V according to the size of the introduction target T. The size of the introduction target T may be, for example, the average size of the introduction target T determined from the image processed by the image processing unit 7. For example, it is assumed that it has a function of calculating an average size of the introduction target T. Further, the size of the introduction target T may be given to the control device 5 from an input means (not shown). The adjustment of the visual field range by the visual field range adjustment means 86 is performed by giving a command to a drive source of a magnification adjustment mechanism (not shown) of the microscope 3, for example. The magnification adjustment mechanism adjusts the magnification by automatic switching of an objective lens (not shown).

As shown in FIG. 1B, the container movement interval determining means 87 is divided into the microscope visual field range V, and when the container position is moved to the adjacent microscope visual field range V, the movement distance is changed to the visual field range adjusting means 86. It is a means which makes it the distance corresponding to the visual field range adjusted by.

Interference avoiding means 79 is means for determining the processing order so that the injection needles 11 do not interfere with each other when the insertion order determining means 10 determines the order of the injection processing of each injection needle into the body T to be introduced. In addition, when the conveyance device 4 of each injection needle 11 is arranged so that mutual interference between the injection needles 11 does not occur, the interference avoiding means 79 does not need to be provided. However, since the plurality of injection needles 11 are operated in parallel within a minute range, the interference avoiding means 79 is configured to perform the injection process on the introduced objects T at positions separated from each other during the parallel operation so as not to cause unexpected interference. Thus, the insertion order determining means 10 may determine the order of the injection processing.

Next, the operation of the microinjection apparatus having the above configuration will be described. The injection process is performed on the entire region in the container 12, but sequentially for each microscope visual field range V. First, the injection process in each microscope field-of-view range V will be described. Positioning of the injection needle 11 for introduction into the introduction target T in the horizontal direction (two orthogonal directions) and the injection direction are all performed by the conveying means 4 provided for each individual injection needle 11. Therefore, the injection process can be performed in parallel by the plurality of injection needles 11 arranged radially around the container 12.

At the time of processing, first, all the introduced objects T in the microscope visual field range V are imaged by the imaging means 2, image processing by the image processing means 7 is performed, the position is determined by the position determination means 8, and the position storage unit 8a. To remember. In addition, although this position determination may be performed collectively for all regions in the container 12 as described later, in this embodiment, after the injection processing to all the introduced objects T in the microscope visual field range V is completed. The container 12 is moved to another close position by the container position adjusting means 1, the position of the introduction target T is recognized by image processing at that position, and the operation of performing the injection process is repeated.

The assigned range E within the microscope visual field V that each injection needle 11 is in charge of is assigned by the assigned range calculation means 9 as determined. In this case, the assigned range calculation unit 9 is configured so that the load of the transfer unit 4 of each injection needle 11 is equalized, in other words, the operation amount of the transfer unit 3 of each injection needle 11 is equalized. The assigned range E is determined, and the injection processing is performed in parallel with each injection needle 11. Thereby, the total processing time is shortened.

Regarding the allocation of the assigned range E, for example, as shown in FIG. 3, the region of the microscope visual field range V is divided into a plurality of assigned ranges E so that the areas handled by the injection needles 11 are substantially equal. As shown in FIG. 4, the region of the microscope visual field range V may be divided into a plurality of assigned ranges E so that the number of introduction bodies T handled by each injection needle 11 becomes substantially equal. In FIG. 4, the dividing line L is slightly rotated with reference to the radiation center O in the microscope visual field range V. This is because when the area of the assigned range E becomes large, the moving distance of the transfer device 4 becomes long and a difference occurs in the processing time, so that there is no difference in the area of the assigned range E as much as possible. However, the example of FIG. 4 does not limit the method of setting the assigned range.

Further, as shown in FIG. 5, the microscope visual field range V may be divided so that the total movement distance in the region (the assigned range E) processed by each injection needle 11 becomes substantially uniform. As in FIG. 4, in the processing to the body to be introduced T, if a difference occurs in the moving distance of the conveying means 4 of each injection needle 11, a difference occurs in the processing time. The insertion order determining means 10 determines the processing order of the introduction target T for each injection needle 11, calculates the total movement distance in that order, and divides the microscope visual field range V so that the total movement distance is substantially uniform. To do.

Further, the assigned range calculation means 9 is not limited to the division method described above with reference to FIGS. 3 to 5, so that the product of the number of processes and the total movement distance in the region (the assigned range E) processed by each injection needle 11 becomes as small as possible. The microscope visual field range V may be divided. The processing time is determined from both the number of processing and the moving distance. In order to make a relative comparison of the processing times of the injection needles 11 simple, the product of both is used.

Further, the microscope visual field range V may be divided so that the processing time in the assigned range E processed by each injection needle 11 becomes substantially uniform. In this case, the assigned range calculation means 9 performs processing time from the waiting time until the injection needle 11 is stopped in the injection target T and the introduction of the introduced substance is completed, and the movement distance of the injection needle 11 in the injection process. And the charge of each injection needle 11 is determined so that the processing time is substantially uniform.

In addition, the assigned range calculation means 9 has a priority condition that no interference between the injection needles 11 occurs when the injection needles 11 are operated in parallel, and the region of the microscope visual field range V under this priority condition. It is good also as what divides. For example, when the interference part I occurs as shown in FIG. 6, the region of the microscope visual field range V is divided as shown in FIG. When the injection needles 11 interfere with each other when the injection needles 11 are operated in parallel, the injection needles 11 cannot be operated in parallel. Therefore, it is a priority condition that interference does not occur, and within the range where this priority condition is satisfied, other conditions, for example, the area in charge is almost equal, or the number of introduced subjects in charge is almost equal. It is practical to divide the region of the range. In that case, an overlapping area may be provided, but it is preferable to make the overlapping area as small as possible. When providing the overlapping portion, the insertion order determining means 10 determines the order of injection into the introduction target T so that the injection needles 11 do not interfere with each other.

Next, the injection process to the entire area in the container 12 will be described. Since the microscope visual field range V is a partial region in the container 12, it is necessary to sequentially move the container 12 and perform the injection process in the entire region in the container 12. In the above-described example, every time the injection process within one microscope field-of-view range V is completed, the container 12 is moved to determine the position of each introducer T within the next microscope field-of-view range V, and the injection process is performed. Although the operation is repeated, the introduced object position may be determined collectively as follows.

For example, when processing the entire region in the container 12, the in-container full range processing command unit 100 first moves the container 12 sequentially according to the microscope visual field range V as shown in FIG. The positions of all the introduced objects T in the container 12 are determined by the position determination means 8 and stored in the position storage unit 8a. The position storage unit 8a stores the position of the introduction target T in association with the microscope visual field range V. Using the information stored in this way, the division of the assigned range E as shown in FIGS. 3 to 6 by the assigned range calculating means 9 and the order of the injection processing by the insertion order determining means 10 are calculated.

Here, the positions of all the introduced objects T in the container 12 are determined in a lump, but depending on the storage capacity of the position storage unit 8a and the processing speed of the assigned range calculation means 9, etc. Up to the position of the object T to be introduced and stored, and during the injection process of the microscope visual field range V, the calculation of the assigned range E of the injection needle 11 of the microscope visual field range V a plurality of times in parallel is performed. The order of the injection processing may be calculated. FIG. 7B shows an example in which the inside of the container 12 is divided into four. A straight line M indicates a boundary line of the division. In this way, by acquiring information on the position of the transfer target T at the movement destination in advance, the calculation of the movement destination can be performed in advance during the injection process of the microscope visual field range V. Can be shortened.

When the container 12 is moved, as shown in FIGS. 8A and 8B, a portion Va overlapping the current microscope field of view V and the next microscope field of view V may be provided. Thereby, it can avoid that the unprocessed to the to-be-introduced body T near the boundary of the microscope visual field range V arises. Here, the overlapping portion may have a constant area as shown by the hatched area in FIG. 8A, and the introduced object T as a reference from the microscope visual field range V as shown in FIG. It may be determined by a predetermined appropriate method, and the movement distance of the introduced object To may be managed as the reference. This also provides the same effect as when the area of the overlapping part is constant. In addition, for the to-be-introduced object To, for example, the to-be-introduced object T at the upper right end in the microscope visual field range V is defined as To.

As described above, the information on the position of the introducer T in the container 12 is obtained, and simultaneously with the injection process, the microscope field-of-view range V is calculated a plurality of times in parallel. It became possible to shorten the operation.

Next, specific examples of the microinjection device main body, which is a portion excluding the control device 5 in the microinjection device, will be described with reference to FIGS. 10 to 14 show a first specific example of the microinjection apparatus main body. Prior to detailed description of the overall configuration of the first specific example, the characteristic configuration and operational effects will be described. In this specific example, the moving bodies 41 and 42 and the advancing / retreating drive means (power units) 45 and 46 are separated for the movement of the transporting means 4 in the two biaxial directions perpendicular to the X-axis direction and the Y-axis direction. By installing the power units 45 and 46 in the fixed part, the mass of the moving body can be reduced, and the conveying means can be driven at high speed. Conventionally, when the injection needle moves in two degrees of freedom in the orthogonal X-axis direction and Y-axis direction in the injection needle transport means, for example, the Y-axis movement mechanism is arranged to be stacked on the X-axis movement mechanism. It was. In this case, since the X moving mechanism drives both the injection needle and the Y-axis moving mechanism, it is difficult to move at high speed. In this embodiment, stacking of such moving mechanisms is avoided, and high-speed movement is realized by weight reduction.

Conventionally, a ball screw or an ultrasonic motor has been used to drive the injection needle, but in this example, the piezoelectric element laminates 19A and 19B are used to move the injection needle 11 in the injection direction (Z-axis direction). By combining a small actuator using (FIG. 13), the transport device 4 that is a manipulator for driving the injection needle 11 is miniaturized. Piezoelectric actuators have high conversion efficiency for converting electrical energy to mechanical energy, and can change the generated displacement relatively easily by changing the applied voltage, and have excellent controllability. In this example, since the crank slider mechanism is used, the displacement of the piezoelectric element laminates 19A and 19B can be enlarged. The transport device 2 has a drive mechanism having at least two degrees of freedom with respect to the introduction target T, so that the tip of the injection needle can be positioned as a single unit. The downsizing of the transfer device 4 makes it possible to arrange a plurality of transfer devices 4 in a limited space around the container 12 containing the introduced object T such as cells. The plurality of transfer devices 4 can simultaneously position the tip of each injection needle 11.

In addition, the positioning control is performed by the control device 5 (FIG. 1A) as described above, but the assigned range E for each injection needle 11 is determined, and the load on the transfer device 4 of each injection needle 11 is equalized. Therefore, the transport device 4 can be further downsized.

By combining a device for reducing the size of the transfer device 4 in the microinjection device main body and a control device for using the plurality of transfer devices 4 with equal loads, all the introduced objects T in the container 12 are provided. In performing the injection process, the processing time can be shortened. That is, according to this embodiment, the transporting means 4 that is a manipulator can be reduced in size, and a plurality of transporting means 4 are arranged around a container 12 such as a petri dish containing an introduced object T such as a cell. be able to. In addition, the tip of the injection needle 11 can be simultaneously positioned with respect to a plurality of introduced bodies T, and injection processing to a large number of introduced bodies T can be executed at a time, thereby enabling high throughput. It becomes. In addition, when performing the injection process, by calculating the assigned range E of each injection needle 11 and the injection order in advance, the waiting time by the calculation can be reduced and the time can be shortened.

The detailed configuration of each specific example of the microinjection apparatus main body will be described. In FIG. 1 (A), the container position adjusting means 1 moves the container mounting table 13 on which the container 12 such as a petri dish for storing the introduction body is held in the horizontal orthogonal biaxial direction as described above. It is means for adjusting the position of the container 12 by moving it, and is constituted by the 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 that is movable in the X-axis direction via the guide 184, and movable base drive mechanisms 186 and 187 for the respective axes that move 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 of FIG. 1A is installed on the upper movable table 185, or the upper movable table 185 itself becomes the container mounting table 13.

In FIG. 1 (A), the conveying means 4 has three degrees of freedom. The moving mechanism that bears one degree of freedom is the first shown in FIG. 10 in one direction (X-axis direction) of two directions (X-axis direction and Y-axis direction) orthogonal to the container mounting table 13 in the horizontal direction. The X-axis moving mechanism 14 moves the injection needle 11 together with one moving body 41. The moving mechanism that bears one other degree of freedom moves the injection needle 11 together with the second moving body 42 shown in FIG. 10 in another direction (Y-axis direction) of the two orthogonal directions in the horizontal direction. This is a Y-axis moving mechanism 15. Still another moving mechanism that bears one degree of freedom is the Z-axis moving mechanism 16 shown in FIG. 10 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.

As shown in FIGS. 11A and 11B, the first moving body 41 constituting the X-axis moving mechanism 14 moves in the linear direction (X through the first guide 43 to the base 40 of the transport device 4. It is installed so that it can move forward and backward in the axial direction. The second moving body 42 constituting the Y-axis moving mechanism 15 has a linear direction (Y-axis direction) orthogonal to the linear direction (X-axis direction) via the second guide 44 on the first moving body 41. ) Can be moved forward and backward. Each of the first moving body 41 and the second moving body 42 has a rectangular plate shape. As shown in FIG. 10, the Z-axis moving mechanism 16 (FIG. 10) is mounted on the second moving body 42. The Z-axis moving mechanism 16 is installed so that the injection angle of the injection needle 11 becomes a predetermined angle inclined downward. Further, the base 40 is advanced and retracted by a transport means retracting mechanism 39 (FIG. 1A).

11A and 11B, first advance / retreat drive means 45 and second advance / retreat drive means 46 for advancing and retreating the first moving body 41 and the second moving body 42 are provided on the base 40, respectively. is set up. The output member 45a of the first advancing / retreating drive means 45 is in a direction (Y-axis direction) orthogonal to the direction (X-axis direction) in which the first moving body 41 can move back and forth with respect to the first moving body 41. On the other hand, it can freely move. The output member 46a of the second advance / retreat drive means 46 is in a direction (X axis direction) orthogonal to the direction (Y axis direction) in which the second movable body 42 can move forward and backward with respect to the second movable body 42. On the other hand, it can freely move.

The first guide 43 includes a rail 43a installed on the base 40, and a guided body 43b such as a linear motion rolling bearing provided on the lower surface of the first moving body 41 and capable of moving forward and backward along the rail 43a. And two are provided in parallel to each other. The second guide 44 includes a rail 44a installed on the first moving body 41 and a linear motion rolling bearing provided on the lower surface of the second track body 42 and capable of moving forward and backward along the rail 44a. It consists of guided bodies 44b, and two are provided in parallel to each other. The rails 43a of the first and second guides 43 and 44 are provided with guide grooves (not shown) along the length direction, and the guided bodies 43b and 44b made of linear motion rolling bearings are provided with the guides. A rolling element (not shown) fitted into the groove so as not to fall off is provided.

The first advance / retreat driving means 45 is means for advancing / retreating the output member 45a in the linear direction by the drive means main body 45b. The drive means main body 45b may be any as long as it can advance and retract the output member 45a. For example, the piezoelectric element laminates 19A and 19B shown in FIG. 13, an ultrasonic motor, or a combination of a motor and a feed screw mechanism such as a ball screw. In this embodiment, the piezoelectric element laminates 19A and 19B shown in FIG. 13 are used. The configuration of FIG. 13 will be described later. The second advancing / retreating drive means 46 is a means for advancing / retreating the output member 46a in the linear direction by the drive means main body 46b, and has the same configuration as the first advancing / retreating drive means 45.

The output members 45a and 46a of the advancing / retreating drive means 45 and 46 are merely in contact with the moving parts 41 and 42 so as to apply a moving force, but from the opposite side as shown in FIGS. 12 (A) and 12 (B). By adopting a structure that is pressed by the elastic bodies 121 and 122, it can be freely driven in both forward and reverse directions. Specifically, the elastic body 121 is disposed between the elastic body support portion 40a provided on the base 40 and the first moving body 41 on the side opposite to the installation side of the first advance / retreat driving means 45. The first movable body 41 is pressed against the output member 45 a of the first advance / retreat driving means 45 by the elastic body 121. Also with respect to the second moving body 42, on the side opposite to the installation side of the second advancing / retreating drive means 46, the elastic body support portion 41a provided on the first moving body 41, the second moving body 42, The elastic body 122 is disposed between the second moving body 42 and the second moving body 42 against the output member 46 a of the second advancing / retreating drive unit 46. The elastic bodies 121 and 122 are, for example, coil springs. The elastic bodies 121 and 122 are preferably springs having a spring constant of 1 N / mm or less.

Here, measures are taken to reduce the coefficient of friction so that the movable bodies 41, 42 and the output members 45a, 45b of the advancing / retracting drive means 45, 46 can move in any direction on the surface perpendicular to the moving direction. It is desirable. As a measure for this, in this example, the tip of the output member 45a of the first advancing / retreating drive means 45 in contact with the first moving body 41 is hemispherical as shown in FIG. Although not shown, the output member 46 of the second advance / retreat drive means 46 also has a spherical end at the output member 46a, like the first advance / retreat drive means 45. Instead of making the front ends of the output members 45a and 46a spherical, the side surfaces of the first moving body 41 or the second moving body 42 with which the front ends of the output members 45a and 46a are in contact with each other slide. It is good also as a semi-cylinder shape extended in a direction, and the front-end | tip part of output member 45a, 46a is good also as a flat surface.

Further, as shown in FIG. 15, a coating 101 for reducing friction may be applied to one or both of the contact portions between the output members 45 a, 46 a of the advance / retreat driving means 45, 46 and the moving bodies 41, 42. good. In the illustrated example, the coating 101 is applied to the output members 45a and 46a. The coating 101 is preferably diamond-like carbon, molybdenum disulfide, fluororesin, or graphite.

In place of the hemispherical or semi-cylindrical shape as in the example of FIG. 15, as shown in FIG. 16, the output members 45a and 46a of the advance / retreat driving means 45 and 16 and the moving bodies 41 and 42 are balls or You may make it contact via rolling elements 49, such as a roller. In the example of FIG. 16, the rolling element 49 is a ball, and is rotatably accommodated in the holding members 45aa and 46aa provided at the tips of the output members 45a and 46a so as to partially protrude.

A specific configuration example of the Z-axis moving mechanism 16 in FIG. 10 will be described later together with FIG.

According to the microinjection apparatus having this configuration, the position of the container 12 containing the introduction target T is adjusted by the container position adjusting means 1, and a planar view image inside the position-adjusted container 12 is picked up by the imaging means 2, By determining the cell position from the image by the position determination means 3, the introduction target T such as a cell in charge of each injection needle 11 is determined. At this time, the position of the introducer T can be recognized with high accuracy by locally enlarging the container 12 with an enlargement lens and performing image processing on the position of the introducer T such as cells. Further, for example, the to-be-introduced body T in charge of each injection needle 11 can be determined from the positional relationship in the image of the to-be-introduced body T such as recognized cells. Thus, after determining the to-be-introduced body T which each injecting needle 11 takes charge of, each injecting needle 11 is moved by several separate conveyance means 4, and the injecting needle 11 is inserted in each to-be-introduced body T, respectively. Then, the introduction substance is introduced into each introduction target T.

Since the conveying means 4 has a drive mechanism with at least two degrees of freedom with respect to the introduction target T, the tip of the injection needle 11 can be positioned as a single unit. Each transport means 4 operates in parallel and repeats the injection operation. After the processing on the cells in the image is completed, the container 12 is moved by the container position adjusting means 1, the cell arrangement at another adjacent position is recognized by image processing, and the injection operation is repeated as described above.

Thus, the introduction substance can be simultaneously introduced into a plurality of recipients T such as cells, and the efficiency of the injection process can be improved. In addition, it is possible to simultaneously introduce a plurality of kinds of introduction substances into the introduction target T by filling at least two kinds of different injection substances for each injection needle and sequentially introducing them into the same introduction target T. It becomes. Therefore, a plurality of types of introduction substances can be introduced without replacing the injection substance in the injection needle 11, and the efficiency of the injection process can be improved.

In particular, as shown in FIGS. 11 to 12, the transport means 4 separates the moving bodies 41, 42 and the advance / retreat drive means 45, 46, and advances / retreats drive means 45, 45 for each degree of freedom of movement in the X axis and Y axis directions. Since both 46 are installed on the base 40 of the transport means 4, the transport means 4 can be reduced in size, and the mass of the moving bodies 41 and 42 can be reduced, so that the transport means 4 can be driven at high speed. Although the advancing / retreating drive means 45 and 46 for each degree of freedom are both installed on the base 40, the second moving body 42 installed on the base 40 via the first moving body 41 is Since the output member 46a of the second advance / retreat driving means 46 is in free contact with the direction orthogonal to the direction in which the advance / retreat movement is possible, the second member 41a regardless of the position where the first moving body 41 has moved. The forward / backward drive of the forward / backward drive means 46 can be transmitted to the second moving body 42.

Moreover, since the conveyance means 4 can be reduced in size, a plurality of conveyance means 4 can be disposed around a container 12 such as a petri dish containing cells to be introduced T as shown in FIGS. it can. Therefore, the tip of the injection needle 11 can be simultaneously positioned with respect to the introducer T, and the injection process to a large number of introducers T can be executed at a time, so that high throughput can be achieved.

Further, when the piezoelectric element laminates 19A and 19B are used for the advance / retreat driving means 45 and 46 of the moving bodies 41 and 42 of the transport means 4, the following effects can be obtained. That is, conventionally, a ball screw or an ultrasonic motor has been used to drive the injection needle. In this embodiment, however, the conveying means 4 for driving the injection needle 11 is combined with a small actuator using a piezoelectric element. We are trying to make it. Piezoelectric actuators have high conversion efficiency for converting electrical energy to mechanical energy, and can change the generated displacement relatively easily by changing the applied voltage, and have excellent controllability.

In the above embodiment, the elastic bodies 121 and 122 are provided as a configuration for maintaining the contact between the output members 45a and 46a of the advance / retreat driving means 45 and 46 and the moving bodies 41 and 42. However, FIGS. Permanent magnets 111 to 114 may be provided as shown in FIG. That is, the first and second permanent magnets 111 and 112 are arranged on the base 40 and the first moving body 41 so that the directions of the magnetic poles facing each other are the same, and the first moving body is generated by the magnetic repulsive force. 41 may be pressed against the output member 45a of the first advance / retreat driving means 45. In addition, the second and fourth permanent magnets 113 and 114 are arranged on the first moving body 41 and the second moving body 42 so that the directions of the magnetic poles facing each other are the same, and the second repulsive force causes the second movement. The moving body 42 may be pressed against the output member 46 a of the second advance / retreat driving means 46.

In the example of FIGS. 18A and 18B, the permanent magnets 111A to 114A are provided so that the magnetic attractive force acts, contrary to the example of FIGS. 17A and 17B. That is, the first and second permanent magnets 111A and 112A are arranged on the base 40 and the first moving body 41 so that the directions of the magnetic poles facing each other are reversed, and the first moving body is generated by the magnetic attraction force. 41 may be pressed against the output member 45a of the first advance / retreat driving means 45. In addition, the second and fourth permanent magnets 113A and 114A are arranged on the first moving body 41 and the second moving body 42 so that the directions of the magnetic poles facing each other are reversed, and the second moving magnet 41 is driven by the magnetic attractive force. The moving body 42 may be pressed against the output member 46 a of the second advance / retreat driving means 46.

In the example of FIGS. 19A and 19B, instead of simply contacting the output member 46a of the second advance / retreat drive means 46 with the second moving body 42, the output member of the second advance / retreat drive means 46 is used. 46a is connected to the second moving body 42 via a third guide 115 so as to be movable in the moving direction (X-axis direction) of the first moving body 41. The third guide 115 has a rolling element (not shown) fitted into a guide rail 115a provided on the second moving body 41 and a guide groove (not shown) provided on the guide rail 115a. And a guided body 115b such as a linear motion rolling bearing which cannot be removed. Other configurations are the same as those shown in FIGS. 12A and 12B. When the third guide 115 is connected in this way, the output member 46a and the moving body 41 can always be connected even during high-speed driving, and the operation is stabilized. The second elastic body 122 provided to face the output member 46a applies a preload to the third guide 115, and ensures further stable operation.

As shown in FIGS. 20A and 20B, a fourth guide 116 may be further provided. That is, the fourth guide 116 is configured so that the output member 46a of the second advance / retreat driving means 46 and the third guide 115 are movable with respect to the moving direction (X-axis direction) of the first moving body 41. Connect through. As with the third guide 115, the fourth guide 116 has a guide rail 116a with a guide groove (not shown) and a rolling element (not shown) fitted into the guide groove of the guide rail 116a. And a guided body 116b such as a linear motion rolling bearing which cannot be removed. The guide rail 116a is attached to the guided body 115b of the third guide 115, and the guided body 116b is attached to the output member 46a. As described above, when the third guide 115 and the fourth guide 116 are provided in two stages, the operation during high-speed driving or the like is further stabilized. In this example, the fifth and the fifth movable members that are movable between the first moving member 41 and the output member 45a of the first driving means 45 in the direction orthogonal to the moving direction of the first moving member 41 are also provided. Six guides 117 and 118 are provided. Similar to the third and fourth guides 115 and 116, the fifth and sixth guides 117 and 118 include a guide rail and a guided body.

FIG. 21 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 end of the pair of piezoelectric element laminates 19 </ b> A is connected to a protrusion 48 b on the one end side of the fixing base 17 in the fastening member 48, and the other end is 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 injection needle individual control unit 88a in the injection needle movement control means 88 of FIG. Further, the position command unit 52 uses the target position determined by the assigned introducer determining unit 87 as the position command based on the position information obtained by the position determination unit 3 in 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.

21 can be used as the first advance / retreat drive means 45 and the second advance / retreat drive means 46 in the X-axis movement mechanism 14 and the Y-axis movement mechanism 15 shown in FIG. In that case, instead of the needle support member 18, an output member 45a is provided as shown in FIG. FIG. 13 shows a specific example of the first advance / retreat driving means 45 in the example of FIGS. 11 (A) and 11 (B). The example shown in FIG. 13 can also be used for the second advance / retreat driving means 46.

Next, each configuration example using piezoelectric elements that can be used as the first advance / retreat drive means 45 and the second advance / retreat drive means 46 in the example of FIGS. 11A and 11B will be described with reference to FIGS. To do.

22, the advance / retreat driving means 45 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 tip 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 portion 26a of the movable piece 26 is bent at the thin portion 26d at the intermediate portion in the longitudinal direction, and the intermediate portion is formed in the direction perpendicular to the longitudinal direction (Y-axis direction) by increasing or decreasing 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 second movement 42 (FIG. 10) that supports the Z-axis moving mechanism 16, whereby the injection needle 11 can move in the Y-axis direction.

2 is applied to the second advancing / retreating drive means 46, the two piezoelectric element laminates 19C and 19D are displaced by a positioning signal which 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 injection needle individual control unit 88a in the injection needle movement control means 88 of FIG. Further, the position command unit 60 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 in FIG. The position command unit 60 may be provided as a part of the assigned introducer determining unit 87.

When applied to the second advance / retreat driving means 46, 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. 23 shows another configuration example that can be used as the first advance / retreat drive means 45 and the second advance / retreat drive means 46. The case of applying to the first advance / retreat drive means 45 will be described. The advance / retreat drive means 45 includes a fixed base 35, a movable piece 36, and two sets of piezoelectric element laminates 19E and 19F serving as drive 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 in the example of 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 example of FIG. 22, the two sets of piezoelectric element laminates 19E and 19F are both laminated piezoelectric elements, and are arranged back and forth so as to be parallel to the main frame portion 35a of the fixed base 35 along the lamination direction. Arranged in parallel. 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 thin frame portion 37b becomes the output member 45a in the example of FIGS. 11 (A) and 11 (B).

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 injection needle individual control unit 88a in the injection needle movement control means 88 of FIG. Further, the position command unit 64 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 in FIG. The position command unit 64 may be provided as a part of the assigned introducer determining unit 87.

FIG. 24 shows another configuration example that can be used as the first advance / retreat drive means 45 and the second advance / retreat drive means 46. Here, a case where the present invention is applied to the second advance / retreat driving means 46 will be described. Similarly to the configuration example of FIG. 22, 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 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 this configuration example, the link mechanism 75 is provided as an expansion mechanism that expands and contracts the piezoelectric element laminates 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.

25A to 25C show various configuration examples of the link mechanism 75. FIG. In the configuration example of FIG. 25A, 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. 25 (B) and 25 (C).

The first and second links 81A and 81B have one end connected to the fixed joint 76 and the first movable joint 77A and the other end connected to each other via the 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, and 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 is 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.

25B, 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. 25A, the movable joint that connects the base ends of the third links 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. In the example of FIG. 25B, the movable portion 82 at the tip of the third link 81C is the 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. 25A and 25B, the angle of the third link 81C can be varied independently even if a dimensional error or thermal deformation of the links 81C, 81B, 81C 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. 25A, 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. 25B, 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.

25C, the 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. 23A and 23B, 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 expansion / contraction direction 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, in the movable joint 77B, it is only necessary to provide one bearing in the axial direction, and the thickness dimension can be reduced to ½ compared to the case of the configuration example in FIG.

24, the link mechanism 75 of the example of FIG. 25C is provided as an enlargement mechanism. Other configurations are the same as the example shown in FIG.

FIG. 26 shows another configuration example that can be used as the first advance / retreat drive means 45 and the second advance / retreat drive means 46. Here, the case where it applies to the 1st advance / retreat drive means 45 is demonstrated. This configuration example also includes a fixed base 83 and two sets of piezoelectric element laminates 19I and 19J serving as a drive source, as in the example of FIG. 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 this example, the first link mechanism 101 is provided as a first expansion mechanism that expands the expansion and contraction of the piezoelectric element laminates 19I and 19J to a displacement in the direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction). In addition, a second link mechanism 102 is provided as a second expansion mechanism that 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 laminates 19I and 19J. ing. As these both link mechanisms 101 and 102, the thing of the same structure as the link mechanism 75 shown to FIG.25 (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 composed of 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 the example shown in FIG.

In the X-axis moving mechanism 14 of FIG. 26, 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. 25C. However, the present invention is not limited to this, and the configuration example shown in FIG. 25B 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.

27, 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 FIGS. 28A and 28B, the link mechanism 75 includes one fixed joint 76, two movable joints 77A and 77B, and two links 81A and 81B. The crank / slider mechanism comprises a fixed joint 76 that moves the displacement of the piezoelectric element in the extending direction via the two movable joints 77A and 77B and the two links 81A and 81B. It is also possible to change in an arbitrary direction on the circumference of the circle and further expand the displacement.

FIG. 28A shows another configuration example of the Y-axis moving mechanism 15. Similarly to the configuration example of FIG. 24, the Y-axis moving mechanism 15 according to this configuration example 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 Side plate 73e which fastens one frame part. 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 this example, a link mechanism 75 is provided as an enlarging mechanism that expands and contracts the piezoelectric element laminates 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 advance / retreat drive means 45 and 46 can be reduced and the size can be reduced.

FIG. 28B shows another configuration example of the X-axis moving mechanism 14. The X-axis moving mechanism 14 in FIG. 28B also has the same configuration as that in FIG. 28A, and the position of the movable joint 77B and the movable portion 82 fixed to the link 81A is changed, so that the X-axis moving mechanism 14 in the X-axis direction is changed. 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. 29 shows an enlarged configuration example of the link mechanism 75 shown in FIGS. 28 (A) and 28 (B). In the configuration example shown in FIGS. 28A and 28B, 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 fixed joints 76 and the movable joints 77A and 77B are joints that respectively constitute rotatable nodes, and the center of rotation is the expansion / contraction direction (X-axis direction) of the piezoelectric element laminates 19G and 19H and It is perpendicular to any direction (Y-axis direction) orthogonal to the expansion / contraction direction (X-axis direction).

The first and second links 81A and 81B have one end connected to the fixed joint 76 and the first movable joint 77A and the other end connected to each other via the 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. 30 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.

As described above, the preferred embodiments have been described with reference to the drawings. However, those skilled in the art will readily assume various changes and modifications within the obvious scope by looking at the present specification. Accordingly, such changes and modifications are to be construed as within the scope of the invention as defined by the appended claims.

DESCRIPTION OF SYMBOLS 1 ... Container position adjustment means 2 ... Imaging means 3 ... Microscope 4 ... Conveyance means 5 ... Control apparatus 7 ... Image processing means 8 ... Position determination means 8a ... Position storage part 9 ... Assigned range calculation means 10 ... Insertion order determination means 11 ... Injection needle 12 ... container 14 ... X-axis moving mechanism 15 ... Y-axis moving mechanism 16 ... Z-axis moving mechanism 19A-19J ... Piezoelectric element laminate 26 ... movable piece (enlargement mechanism)
36 ... movable piece (first enlargement mechanism)
37 ... Movable frame (second enlargement mechanism)
40 ... Base 41 ... First moving body 42 ... Second moving body 43 ... First guide 44 ... Second guide 45 ... First forward / backward driving means 45a ... Output member 46 ... Second forward / backward driving means 46a ... output member 54 ... vibration driving means 75 ... link mechanism (enlargement mechanism)
76 ... fixed joints 77A, 77B ... movable joints 81A-81C ... link 79 ... interference avoiding means 86 ... visual field range adjusting means 88 ... injection needle movement control means 89 ... container movement control means 100 ... in-container full range processing means 101 ... first 1 link mechanism (first expansion mechanism)
102 ... Second link mechanism (second enlargement mechanism)
106 ... fixed joints 107A, 107B ... movable joints 108A, 108B ... link 115 ... third guide 114 ... fourth guide 121 ... first elastic body 122 ... second elastic body E ... responsible range V ... microscope field of view range Va ... overlapping portion T ... introduced object To ... standard introduced object M ... division boundary L in container ... dividing line O in microscope field of view range ... center in microscope field of view

Claims (28)

1. A microinjection apparatus for performing an injection process for introducing an introduction substance into an introduction body by inserting an injection needle filled with the introduction substance into the introduction body,
   Container position adjusting means for adjusting the position of the container containing the introduced body;
   A plurality of conveying means capable of individually moving a plurality of injection needles in at least a two-dimensional direction with respect to the introduction body inside the container whose position is adjusted by the container position adjusting means;
   Imaging means for imaging the inside of the container whose position is adjusted by the container position adjusting means through a microscope in a microscope visual field range;
   Position determining means for determining the position of each introduced object in the microscope visual field range from the image obtained by the imaging means;
   From the information on the position of each introducer determined by the position determination means, the load on the conveying means of each injection needle is equalized in accordance with a predetermined rule within the microscope field of view that each injection needle is responsible for. Responsible range calculation means for determining the range of
   For each introducer in the assigned range determined by the assigned range calculating means, an injection for causing the respective conveying means to move the injection needle from the information on the position of each introduced object determined by the position determining means A microinjection device comprising needle movement control means.
2. 2. The microinjection apparatus according to claim 1, wherein the assigned range calculation means divides the region of the microscope visual field range so that the areas handled by the injection needles are substantially equal.
3. 2. The microinjection apparatus according to claim 1, wherein the assigned range calculation means divides the region of the microscope visual field range so that the number of introduced objects handled by each injection needle is substantially equal.
4. The injection needle movement control means according to claim 1, wherein the injection needle movement control means is configured so that the total movement distance of the injection needle in the operation of the injection processing to each of the introduction bodies in charge is minimized for each injection needle. A microinjection apparatus provided with an insertion order determining means for determining the order of injection processing.
5. In Claim 1, for each individual injection needle, the insertion for determining the order of the injection process of the introduction body so that the total movement distance of the injection needle in the operation of the injection process to each introduction body in charge is the shortest A microinjection apparatus provided with order determination means, wherein the assigned range calculation means divides the region of the microscope visual field range so that the total movement distances of the individual injection needles are substantially equal.
6. The injection needle movement control means according to claim 1, wherein the injection needle movement control means is configured so that the total movement distance of the injection needle in the operation of the injection processing to each of the introduction bodies in charge is minimized for each injection needle. An insertion order determining means for determining the order of the injection processing is provided, and the assigned range calculating means divides the region of the microscope visual field range so that the product of the total moving distance and the number of processed individual injection needles is substantially equal. Microinjection device.
7. The injection needle movement control means according to claim 1, wherein the injection needle movement control means is configured so that the total movement distance of the injection needle in the operation of the injection processing to each of the introduction bodies in charge is minimized for each injection needle. An insertion order determining means for determining the order of the injection processing is provided, and the assigned range calculating means stops the introduction time of the injection needle and introduction of the introduction substance while the injection needle is inserted into the introducer. A microinjection apparatus that calculates an injection time and divides the region of the microscope visual field range so that a processing time that is the sum of the injection time and the movement time is substantially equal.
8. In Claim 1, the said charge range calculating means makes it a priority condition that interference between injection needles does not occur when each injection needle is operated in parallel, and the region of the microscope visual field range under this priority condition. Microinjection device to divide.
9. 2. The microinjection apparatus according to claim 1, further comprising interference avoiding means for determining an order of injection processing of each injection needle to the introduction target so that the injection needles do not interfere with each other.
10. 2. The microinjection apparatus according to claim 1, further comprising a field-of-view range adjusting unit that adjusts the microscope field-of-view range in accordance with the size of the introduction target.
11. 2. The container position adjusting means according to claim 1, wherein the container position adjusting means sequentially moves the position of the container in accordance with a microscope visual field range, and performs the injection processing of the injection needle in the entire region in the container. A microinjection apparatus provided with a full range processing means in a container for controlling a transport means of an injection needle.
12. 12. The in-container full range processing means according to claim 11, wherein after the injection processing is performed on all the introduction objects in the microscope visual field range, the container is sequentially moved by the container position adjusting means in accordance with the microscope visual field range. A microinjection device that performs an injection process on an introduced object in a container at a position after each movement.
13. In Claim 11, when providing the in-container full range processing means, it comprises an insertion order determining means for determining the order of the injection processing of each injection needle from a predetermined rule, the in-container full range processing means, The position of the container is sequentially moved by the container position adjusting means according to the microscope visual field range, and the position of the introduced object in each microscope visual field range of the plurality of movement destinations is determined by the position determining means before the injection process. A microinjection apparatus that makes a determination, causes the assigned range calculation means to determine the range assigned to each injection needle, and causes the insertion order determination means to determine the order of injection processing.
14. 12. The method according to claim 11, further comprising insertion order determining means for determining the order of injection processing of each injection needle from a predetermined rule, wherein the entire range processing means in the container is controlled by the container position adjusting means according to the microscope visual field range. The positions of all the introduced objects in the container are sequentially moved by the position determination means before the injection process, and the range for each injection needle is assigned to the assigned range calculation means. A microinjection apparatus that determines the order of injection processing by the insertion order determination means.
15. In Claim 11, the said whole range processing means in a container moves the position of a container sequentially by the said container position adjustment means according to a microscope visual field range, and the position of the introduce | transduced body of the present microscope visual field range by a position determination means. A microinjection apparatus that causes the injection process to the injection target movement control means to perform the injection process to each introduced object in the microscope visual field range a plurality of times before the determination.
16. 12. The microinjection apparatus according to claim 11, wherein when the container position adjusting means sequentially moves the position of the container, the entire range processing means in the container moves so as to have an overlapping portion in the microscope visual field range.
17. 17. The microinjection apparatus according to claim 16, wherein the in-container full range processing means makes the overlapping portions have the same area.
18. 17. The entire range processing means in a container according to claim 16, wherein when the position of the container is sequentially moved by the container position adjusting means, an object to be introduced is determined as a reference within the microscope visual field range according to a predetermined rule. And a microinjection apparatus for moving the region including the introduced body so as to overlap each other.
19. 2. The conveying means according to claim 1, further comprising: a moving body that is installed so as to be movable forward and backward; and an advancing / retreating driving means that moves the moving body, and the advancing / retreating driving means includes a plurality of piezoelectric elements stacked as a drive source. And a microinjection device having a piezoelectric element laminate that expands and contracts in the lamination direction.
20. 20. The advancing / retreating drive means using the piezoelectric element laminate in the transport means as a drive source according to claim 19, wherein the piezoelectric element laminates are arranged in parallel, and the plurality of piezoelectric element laminates are stretched in the extension direction via a coupling member. Microinjection device connected in series.
21. 21. The microinjection apparatus according to claim 20, wherein the advancing / retreating drive unit using the piezoelectric element laminate in the transport unit as a drive source has an expansion mechanism that expands and contracts the piezoelectric element laminate to a displacement in a direction perpendicular to the expansion / contraction direction. .
22. The microinjection device according to claim 21, wherein the expansion mechanism is a link mechanism.
23. 21. The microinjection apparatus according to claim 20, wherein the advancing / retreating drive unit using the piezoelectric element laminate in the transport unit as a drive source has an expansion mechanism that expands and contracts the piezoelectric element laminate to displacement in a direction parallel to the expansion / contraction direction. .
24. 24. The microinjection device according to claim 23, wherein the expansion mechanism is a link mechanism.
25. 23. The link mechanism according to claim 22, wherein the link mechanism includes a crank / slider mechanism including one fixed joint, two movable joints, and two links. A microinjection device that can be converted into an arbitrary direction on the circumference of the fixed joint and further expand the displacement through the two movable joints and the two links.
26. 25. The link mechanism according to claim 24, wherein the link mechanism has a crank-slider mechanism including one fixed joint, two movable joints, and two links, and the crank-slider mechanism is configured to reduce displacement in the extension direction of the piezoelectric element. A microinjection device that can be converted into an arbitrary direction on the circumference of the fixed joint and further expand the displacement through the two movable joints and the two links.
27. 2. The conveying means according to claim 1, having at least two degrees of freedom in which at least the first moving body and the second moving body can advance and retreat in directions orthogonal to each other, and the injection needle is attached to the second moving body. The first moving body is installed on the base through the first guide so as to be movable forward and backward in a linear direction, and the second moving body is disposed on the first moving body via the second guide. A moving body is installed so as to be able to advance and retreat in a direction orthogonal to the linear direction, and the first advancing / retreating driving means and the second advancing / retreating driving means for advancing and retracting the first moving body and the second moving body, respectively, A microinjection device installed on a table and in which the output member of the second advancing / retreating drive unit is movably connected to or in contact with a direction perpendicular to the direction in which the second moving body can move back and forth. .
28. 2. The microinjection apparatus according to claim 1, wherein the introducer is a cell and the introduced substance is a gene regulatory factor.

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