Suction and discharge device and suction and discharge method

Abstract

To provide a suction and discharge device and method capable of performing both an agitation process and a suction and discharge process stably.SOLUTION: A suction and discharge device 1 capable of moving a fluid from a first container 31 to a second container 32, includes: a first pipette 21a capable of agitating the fluid stored in the first container 31 by suctioning and discharging it; a second pipette 21b capable of suctioning the agitated fluid from the first container 31 and discharging the suctioned fluid into the second container 32 to be arranged; a transport unit 20 capable of changing a relative position of the second pipette 21b with respect to the first container 31 and the second container 32; and a control unit 50 capable of controlling the first pipette 21a, the second pipette 21b, and the transport unit 20.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a suction and discharge device and a suction and discharge method. [Background technology]

[0002] Traditionally, research and development in the field of regenerative medicine has typically involved experiments being performed manually, and there is a demand for automation of each experimental device to make experiments more efficient and reduce the labor required.

[0003] In cell culture experiments, one of the experimental tasks that is generally performed manually is "cell seeding," which involves moving cells into a culture vessel for cell culture. Pipetting is one of the tasks required for cell seeding. In a cell culture method described in Patent Document 1, pipetting is performed to aspirate cell clusters from cell culture carriers.

[0004] To seed cells, a pipette is immersed in a cell suspension, and then agitation (pipetting) is performed by repeatedly suctioning and discharging the precipitated cells to make a uniform suspension, followed by an aspiration and discharge process in which the cells are sucked up using the pipette and discharged into a culture vessel at the destination. Since the amount of cells discharged in the aspiration and discharge process is very small, a high-precision pipette capable of sucking up and discharging a few μL is often used. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent No. 5252411 Summary of the Invention [Problem to be solved by the invention]

[0006] However, if a pipette with an accuracy of about several μL used in the suction and discharge process to move cells is used in the stirring process, the capacity of the pipette is small and there is a risk that the cells contained in the cell suspension will not disperse properly. If the cells do not disperse properly and settle to the bottom, the number of cells in the aspirated solution will be sparse, causing the problem that the cell concentration will not be constant.

[0007] In consideration of the above circumstances, an object of the present invention is to provide a suction and discharge device and a suction and discharge method that can stably carry out a stirring step and a suction and discharge step. [Means for solving the problem]

[0008] In order to solve the above problems, the present invention proposes the following means. The suction and discharge device of the present invention is an suction and discharge device capable of moving a fluid from a first container to a second container, and includes a first pipette capable of stirring the fluid contained in the first container by aspirating and discharging it, a second pipette capable of aspirating the stirred fluid from the first container and discharging the aspirated fluid into the second container, a transport unit capable of changing the relative position of the second pipette with respect to the first container and the second container, and a control unit capable of controlling the first pipette, the second pipette, and the transport unit.

[0009] The aspirating and discharging method of the present invention is an aspirating and discharging method for moving a fluid contained in a first container to a second container, and includes a stirring step of aspirating and discharging the fluid contained in the first container with a first pipette to stir it, an aspirating step of aspirating the stirred fluid from the first container with a second pipette, and a discharging step of discharging the fluid aspirated by the second pipette into the second container. Effect of the Invention

[0010] According to the suction and discharge device of the present invention, it is possible to provide a suction and discharge device and a suction and discharge method that are capable of stably carrying out the stirring step and the suction and discharge step. [Brief description of the drawings]

[0011] [Figure 1] 1 is a perspective view showing a suction and discharge device according to an embodiment of the present invention; [Diagram 2] FIG. 2 is a front view showing the suction and discharge device. [Diagram 3] FIG. 2 is a side view showing the suction and discharge device. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. Fig. 1 is a perspective view showing the suction and discharge device 1 according to this embodiment, Fig. 2 is a front view showing the suction and discharge device 1, Fig. 3 is a side view showing the suction and discharge device 1.

[0013] The suction and discharge device 1 includes a base unit 10, a transport unit 20, a pipette 21, a container unit 30, an exchange unit 40, and a control unit 50. The suction and discharge device 1 is a device capable of transferring a fluid from a first container 31 included in the container unit 30 to a second container 32. The container unit 30 will be described in detail later.

[0014] 1, in this embodiment, the vertical direction in the suction and discharge device 1 is defined as the "up-down direction V," the vertically upward direction is defined as the "upward UP" in the vertical direction V, and the vertically downward direction is defined as the "downward LO" in the vertical direction V. In addition, among the horizontal directions perpendicular to the vertical direction V, the direction in which the first container 31 and the second container 32 are arranged side by side is defined as the "left-right direction H," one side in the left-right direction H is defined as the "leftward LT," and the other side is defined as the "rightward RT" in the left-right direction H. In addition, the direction perpendicular to the vertical direction V and the left-right direction H is defined as the "front-rear direction D," one side in the front-rear direction D is defined as the "forward FR," and the other side is defined as the "rearward RR" in the front-rear direction D.

[0015] The base 10 is, for example, a plate-shaped member, and is a base portion on which each component of the suction and discharge device 1 is installed. The transport unit 20 is a transport device capable of transporting the pipette 21. The transport unit 20 will be described in detail later.

[0016] The pipette 21 is an electric pipette having a tip 211 at its tip, and is capable of aspirating a fluid from the tip of the tip 211. In addition, the pipette 21 is capable of aspirating a fluid and then discharging the aspirated fluid from the tip of the tip 211.

[0017] The pipette 21 includes, for example, a pump device such as a syringe pump inside, and operates the pump to aspirate or discharge a fluid from the tip 211. The pump device included in the pipette 21 adjusts the internal pressure of the pipette 21, for example, to create a positive or negative pressure inside the pipette 21, thereby enabling aspirating or discharging a fluid. The pipette 21 may be connected to an external pump device, and configured so that the internal pressure can be adjusted by the external pump device.

[0018] The pipette 21 includes a first pipette 21a and a second pipette 21b. The first pipette 21a is an electric pipette having a first tip 211a at its tip. The second pipette 21b is an electric pipette having a second tip 211b at its tip. As shown in Figures 1 and 2, the first pipette 21a and the second pipette 21b are arranged side by side in the left-right direction H. The first pipette 21a is arranged to the left LT of the second pipette 21b.

[0019] The container part 30 includes a first container 31, a second container 32, a container part main body 33, and a cooling part .

[0020] The first container 31 is a container that contains a fluid, for example, a tube used for cell culture. The first container 31 contains a cell suspension containing cells and a culture medium. The cells contained in the cell suspension contained in the first container 31 are, for example, mouse skull-derived fibroblasts (MC3T3-E1) and human hepatic cancer-derived cell lines (HuH-7 and HepG2). The first container 31 has an opening on the upper side UP.

[0021] The second container 32 is a dish or petri dish used for cell culture. A methylcellulose liquid (3% methylcellulose medium) is contained in the second container 32. The second container 32 has an opening on the upper side UP.

[0022] The container body 33 is a base of the container 30, and is a box placed on the upper UP surface of the base 10. The first container 31 and the second container 32 are provided in the container body 33.

[0023] A recessed portion recessed downward LO is provided on the upper surface UP of the container body 33. The first container 31 and the second container 32 are provided in the recessed portion of the container body 33. Therefore, the relative positions of the first container 31 and the second container 32 with respect to the container body 33 in the horizontal direction (left-right direction H and front-back direction D) hardly shift.

[0024] The cooling unit 34 is accommodated inside the container body 33. The cooling unit 34 is, for example, a cooling agent containing liquid or ice whose temperature is lower than the outside air temperature. Here, the outside air temperature refers to the air temperature of the space in which the suction and discharge device 1 is provided. The container body 33 is formed of a metal or the like having high thermal conductivity. Therefore, by providing the cooling unit 34 inside the container body 33, the container body 33 is cooled by the cooling unit 34.

[0025] Furthermore, by cooling the container body 33 by the cooling unit 34, the first container 31 and the second container 32 provided on the upper UP surface of the container body 33 are also cooled. In particular, since the second container 32 contains a methylcellulose liquid, by cooling the second container 32 by the cooling unit 34, the fluidity of the methylcellulose liquid contained in the second container 32 can be improved.

[0026] Next, the configuration of the transport unit 20 will be described. The conveying section 20 includes a vertical conveying section 22, a first horizontal conveying section 23, and a second horizontal conveying section 24.

[0027] The vertical transport unit 22 is a moving mechanism capable of moving the pipette 21 in the vertical direction V. The vertical transport unit 22 has a first vertical transport unit 22a and a second vertical transport unit 22b.

[0028] The first vertical transport part 22a is capable of moving the first pipette 21a in the vertical direction V. The second vertical transport unit 22b is capable of moving the second pipette 21b in the vertical direction V.

[0029] The vertical conveying unit 22 is a linear motion mechanism constituted by, for example, a ball screw and an electric motor, etc. The vertical conveying unit 22 can move the pipette 21 connected to the vertical conveying unit 22 in the vertical direction V by being driven by the electric motor. In addition, the first vertical conveying unit 22a and the second vertical conveying unit 22b can be driven independently of each other.

[0030] The first horizontal transport unit 23 is a movement mechanism capable of moving the vertical transport unit 22 in the front-rear direction D. The first horizontal transport unit 23 is capable of moving the vertical transport unit 22 and the pipette 21 in the front-rear direction D.

[0031] The first horizontal transport unit 23 is, for example, a linear motion mechanism constituted by a ball screw, an electric motor, etc. The first horizontal transport unit 23 can move the vertical transport unit 22 and the pipette 21 in the front-rear direction D by being driven by the electric motor.

[0032] The second horizontal transport unit 24 is a moving mechanism capable of moving the first horizontal transport unit 23 in the left-right direction H. The second horizontal transport unit 24 is capable of moving the first horizontal transport unit 23, the vertical transport units 22, and the pipette 21 in the left-right direction H.

[0033] The second horizontal conveying unit 24 is, for example, a linear motion mechanism including a ball screw and an electric motor, etc. The second horizontal conveying unit 24 can move the first horizontal conveying unit 23, the vertical conveying unit 22, and the pipette 21 in the left-right direction H by being driven by the electric motor.

[0034] The second horizontal transport unit 24 is fixed to the upper UP surface of the base 10. Similarly, the container body 33 is fixed to the upper UP surface of the base 10. Therefore, when the pipette 21 moves relative to the base 10 by the vertical transport units 22, the first horizontal transport unit 23, and the second horizontal transport unit 24, the pipette 21 moves relative to the container body 33.

[0035] Since the first container 31 and the second container 32 are provided on the upper UP surface of the container body 33, when the pipette 21 moves relative to the container body 33, the pipette 21 moves relative to the first container 31 and the second container 32.

[0036] The replacement unit 40 is a tip replacement device capable of replacing the tip 211 of the pipette 21. The replacement unit 40 includes a tip removal unit 41 and a tip attachment unit .

[0037] 1, the tip removing part 41 includes a tip removing plate 41a that is provided at an opening portion that opens upward UP of the tip removing part 41 and has a notch portion that opens forward FR. A method for removing the tip 211 by the tip removing part 41 will be described later.

[0038] The tip mounting part 42 is provided near the tip removal part 41 and holds tips having the same shape as the first tip 211a and the second tip 211b. The method of mounting the tip 211 by the tip mounting part 42 will be described later. The tip removal part 41 and the tip mounting part 42 are fixed to the upper UP surface of the base part 10.

[0039] The control unit 50 is a control device capable of controlling a part or the whole of the aspirating and discharging device 1. The control unit 50 is connected to at least the transport unit 20 and the pipette 21 by wire or wirelessly, and is capable of controlling the transport unit 20 and the pipette 21. The control unit 50 may be included in the transport unit 20 or the pipette 21. The control unit 50 is omitted in FIG. 3.

[0040] The control unit 50 is, for example, a program executable device (computer) equipped with a processor, a memory, a storage unit, and the like. Each function of the control unit 50 is realized by, for example, one or more processors such as a CPU (Central Processing Unit) or a GPU (Graphics Processing Unit) executing a program stored in a program memory. However, all or part of these functions may be realized by hardware (e.g., a circuit unit; circuitry) such as an LSI (Large Scale Integration), an ASIC (Application Specific Integrated Circuit), an FPGA (Field-Programmable Gate Array), or a PLD (Programmable Logic Device). In addition, all or part of the above functions may be realized by a combination of software and hardware. The storage unit is realized by a flash memory, an EEPROM (Electrically Erasable Programmable Read-Only Memory), a ROM (Read-Only Memory), a RAM (Random Access Memory), or the like.

[0041] Next, a method for sucking and discharging a fluid by the sucking and discharging device 1 will be described.

[0042] The aspirating and discharging device 1 performs an agitation step of agitating the fluid contained in the first container 31. Here, the fluid contained in the first container 31 is a cell suspension containing cells and a culture fluid. In the agitation step, first, the control unit 50 controls the transport unit 20 to move the first pipette 21a above the first container 31 UP.

[0043] The control unit 50 moves the first pipette 21a to the upper UP of the first container 31, and then controls the transport unit 20 to move the first pipette 21a downward LO and immerse the tip of the first tip 211a into the fluid in the first container 31 through the opening at the upper UP of the first container 31.

[0044] After immersing the first tip 211a in the fluid, the control unit 50 controls the first pipette 21a to repeatedly perform the operation of aspirating the fluid from the tip of the first tip 211a and discharging the aspirated fluid from the tip of the first tip 211a into the first container 31.

[0045] The first pipette 21a agitates the fluid by repeatedly aspirating and discharging the fluid in the first container 31. Specifically, when the fluid is a cell suspension containing cells and a culture solution, the first pipette 21a repeatedly aspirates and discharges the cell suspension, performing pipetting so that the cells are uniformly dispersed in the culture solution.

[0046] Next, the aspirating and discharging device 1 performs an aspirating and discharging step using the second pipette 21b. In the aspirating and discharging step, first, the aspirating and discharging device 1 performs an aspirating step using the second pipette 21b.

[0047] In the aspiration step, first, the control unit 50 controls the transport unit 20 to move the second pipette 21b above the first container 31 UP.

[0048] The control unit 50 moves the second pipette 21b to the upper UP of the first container 31, and then controls the transport unit 20 to move the second pipette 21b to the lower LO and immerse the tip of the second tip 211b into the fluid in the first container 31 through the opening at the upper UP of the first container 31.

[0049] After immersing the second tip 211b in the fluid, the control unit 50 controls the second pipette 21b to aspirate the fluid from the tip of the second tip 211b. At this time, since the fluid in the first container 31 has been stirred by the stirring process, the second pipette 21b aspirates a cell suspension in which the cells are uniformly dispersed in the culture solution. The fluid aspirated by the second pipette 21b remains inside the second tip 211b.

[0050] In the aspirating and discharging step, the aspirating and discharging device 1 then performs a discharging step using the second pipette 21b. In the discharging step, the control unit 50 controls the transport unit 20 to move the second pipette 21b to the upper side UP of the second container 32. At this time, the fluid aspirated in the aspirating step is contained inside the second tip 211b.

[0051] Next, the control unit 50 controls the transport unit 20 and the second pipette 21b to bring the tip of the second tip 211b close to the methylcellulose liquid contained in the second container 32 and eject the fluid aspirated in the suction process onto the surface (upward UP surface) of the methylcellulose liquid.

[0052] In the discharging step, the second tip 211b discharges the fluid multiple times into the second container 32. When discharging multiple times, the control unit 50 controls the transport unit 20 to change the relative position of the second tip 211b with respect to the second container 32, and causes the second pipette 21b to discharge the fluid so as not to overlap with the discharged fluid. The aspirating and discharging device 1 repeats the discharge operation of the second tip 211b and the operation of changing the position of the second tip 211b by the transport unit 20, thereby seeding the fluid into the second container 32.

[0053] Here, for example, the amount of fluid discharged by the second pipette 21b in the discharge step is 0.5 μL, and the amount of fluid aspirated by the second pipette 21b in the aspirating step is 10.0 μL.

[0054] The aspirating and discharging device 1 repeats the discharging operation of the second tip 211b and the operation of changing the position relative to the second container 32, and deposits 0.5 μL of the 10.0 μL of fluid aspirated in the suction step into the second container 32. That is, the aspirating and discharging device 1 divides the cell suspension into 20 clumps (cell clumps) and deposits them on the surface of the methylcellulose liquid in the second container 32.

[0055] The cell masses that the suction and discharge device 1 creates on the surface of the methylcellulose liquid in the second container 32 are, for example, spheroids. The diameter of the spheroids created by the suction and discharge device 1 is, for example, 100 μm or more and 700 μm or less, and preferably 100 μm or more and 200 μm or less. In the above-mentioned stirring step, the suction and discharge device 1 adjusts the concentration of the cell suspension contained in the first container 31 so that, for example, the number of cells per spheroid is 2000.

[0056] For example, the volume of fluid that the second pipette 21b can aspirate is 0.5 μL or more and 20.0 μL or less. Also, the volume of fluid that the first pipette 21a can aspirate is 50.0 μL or more and 1000.0 μL or less. In other words, the volume of fluid that the first pipette 21a can aspirate is greater than the volume of fluid that the second pipette 21b can aspirate.

[0057] The second pipette 21b is a pipette with a precision capable of aspirating and discharging about several μL in order to aspirate and discharge cells in the aspirating and discharging step. The first pipette 21a aspirates and discharges the cell suspension in the stirring step, and stirs the cell suspension. If a pipette with a capacity of about several μL like the second pipette 21b is used in the stirring step, the capacity that can be aspirated and discharged is small, and there is a risk that the cell suspension will not be sufficiently stirred.

[0058] By performing the stirring step using the first pipette 21a, which can aspirate a larger volume of fluid than the second pipette 21b, the first pipette 21a can sufficiently stir the fluid even if the volume of the second pipette 21b is only a few μL. In addition, since the second pipette 21b has a volume suitable for the cells contained in the cell suspension, the aspirating and discharging step using the second pipette 21b can be stably performed.

[0059] The volume of fluid that the pipette 21 can aspirate is determined, for example, by the volume of the tip 211. For this reason, the first tip 211a of the first pipette 21a has a larger volume than the second tip 211b of the second pipette 21b.

[0060] The suction and discharge device 1 may repeat the above-mentioned stirring step and suction and discharge step (suction step and discharge step) to seed all of the fluid contained in the first container 31 into the second container 32. Alternatively, the fluid may be seeded into a plurality of second containers 32.

[0061] The aspirating and discharging device 1 may perform a step of replacing the tip 211 after performing the aspirating and discharging step using the second pipette 21b.

[0062] In the replacement process, the control unit 50 controls the transport unit 20 to move the pipette 21 to the upper UP of the opening of the tip removal unit 41. Next, the control unit 50 controls the transport unit 20 to move the pipette 21 to the lower LO and insert the tip 211 into the opening of the tip removal unit 41.

[0063] Here, a horizontally concave groove is formed at the base (upward end UP) of tip 211. Control unit 50 controls transport unit 20 to move pipette 21 backward RR and engage the groove at the base of tip 211 with the notch of tip removal plate 41a. Control unit 50 controls transport unit 20 to move pipette 21 upward UP with the notch of tip removal plate 41a engaged with the groove at the base of tip 211, and removes tip 211 from pipette 21.

[0064] At this time, the tip removal unit 41 may have a tip collection box capable of storing the tip 211 removed from the pipette 21. For example, a tip collection box is provided below the tip removal plate 41a (LO), and the tip 211 removed by the tip removal plate 41a falls into the tip collection box and is stored therein.

[0065] Next, the control unit 50 controls the transport unit 20 to move the pipette 21 to the upper side UP of the tip mounting unit 42. At this time, the tip 211 is not attached to the tip of the pipette 21. In addition, an unused tip 211 is held in the tip mounting unit 42.

[0066] The control unit 50 controls the transport unit 20 to move the pipette 21 downward LO and connect the tip of the pipette 21 from upward UP to an unused tip 211 held in the tip attachment unit 42, thereby attaching a new tip 211 to the pipette 21. For example, even when the stirring step and the aspirating and discharging step are performed consecutively on a plurality of first containers 31 containing different fluids, the aspirating and discharging device 1 can suppress the fluid attached to the tip 211 from mixing with a different fluid by performing the tip 211 replacement step.

[0067] The first tip 211a and the second tip 211b have different capacities and therefore different sizes. Therefore, the tip removal plate 41a is provided with a plurality of cutouts corresponding to the sizes of the first tip 211a and the second tip 211b. The tip mounting section 42 holds the unused first tip 211a and the unused second tip 211b.

[0068] The suction and discharge device 1 of this embodiment is equipped with a first pipette 21a capable of stirring the fluid contained in the first container 31 by aspirating and discharging it, a second pipette 21b capable of aspirating the stirred fluid from the first container 31 and discharging the aspirated fluid into the second container 32, a transport unit 20 capable of changing the relative position of the second pipette 21b with respect to the first container 31 and the second container 32, and a control unit 50 capable of controlling the first pipette 21a, the second pipette 21b, and the transport unit 20.

[0069] Therefore, by using a pipette (here, first pipette 21a) suitable for the stirring process of the fluid contained in the first container 31 and a pipette (here, second pipette 21b) suitable for the suction and discharge processes for moving the fluid from the first container 31 to the second container 32, it is possible to provide an suction and discharge device 1 and an suction and discharge method using the suction and discharge device 1 that can stably perform the stirring process and suction and discharge processes.

[0070] Although one embodiment of the present invention has been described in detail above with reference to the drawings, the specific configuration is not limited to this embodiment, and design modifications and the like within the scope of the present invention are also included. In addition, the components shown in the above-mentioned embodiment and the modified examples shown below can be appropriately combined to form a configuration.

[0071] (Variation 1) In the above embodiment, the transport unit 20 can move the pipette 21 relative to the container unit 30 fixed to the base unit 10, but the mode of the transport unit is not limited to this. The transport unit may change the position of the container unit. For example, the transport unit may be a movable stage provided on the upper UP surface of the base unit and capable of moving the container unit. The transport unit may also include a movable stage capable of moving the container unit horizontally and a transport mechanism capable of moving the pipette vertically. [Explanation of symbols]

[0072] 1 Suction and discharge device 10 Base 20 Conveyor section 21 Pipettes 21a First pipette 21b Second pipette 211 Chip 211a First Chip 211b Second Chip 22 Upper and lower conveying section 22a First vertical conveying section 22b Second upper and lower conveying section 23 First horizontal conveyor section 24 Second horizontal conveyor section 30 Container section 31 First container 32 Second container 33 Container body 34 Cooling section 40 Replacement Part 41 Chip removal section 42 Tip mounting part 50 Control section V vertical direction UP upward LO Downward H Horizontal direction (left - right direction) LT Left side RT Right side D Front - back direction (horizontal direction) FR Front side RR Rear side

Claims

1. A suction and discharge device capable of transferring a fluid from a first container to a second container, a first pipette capable of stirring the fluid contained in the first container by aspirating and discharging the fluid; a second pipette capable of aspirating the stirred fluid from the first container and dispensing the aspirated fluid into the second container; a conveying unit capable of changing a relative position of the second pipette with respect to the first container and the second container; a control unit capable of controlling the first pipette, the second pipette, and the transport unit; Equipped with Suction and discharge device.

2. The volume of the fluid that the first pipette can aspirate is greater than the volume of the fluid that the second pipette can aspirate. The suction and discharge device according to claim 1 .

3. The volume of the fluid that the first pipette can aspirate is 50.0 μL or more and 1000.0 μL or less, The volume of the fluid that the second pipette can aspirate is 0.1 μL or more and 20.0 μL or less. The suction and discharge device according to claim 2 .

4. The second pipette dispenses the fluid into the second container containing methylcellulose liquid. The suction and discharge device according to claim 1 .

5. A cooling unit that cools the second container is provided. The suction and discharge device according to claim 4.

6. The conveying unit is When the second pipette aspirates the fluid, the second pipette is moved above the first container; moving the second pipette above the second container when the second pipette dispenses the fluid; The suction and discharge device according to any one of claims 1 to 5.

7. The second pipette deposits the fluid aspirated from the first container into the second container in a plurality of portions; The transport unit moves the second pipette so that the fluids placed by the second pipette in the second container do not overlap. The suction and discharge device according to claim 6.

8. A method for aspirating and discharging a fluid contained in a first container to a second container, comprising the steps of: a stirring step of aspirating and discharging the fluid contained in the first container with a first pipette; an aspirating step of aspirating the stirred fluid from the first container with a second pipette; a discharge step of discharging the fluid aspirated by the second pipette into the second container; Equipped with Suction and discharge method.

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