JP2018140375A - Agitation device, agitation method, cell culturing method and reaction acceleration method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a stirring device and a stirring method capable of efficiently and quickly stirring a fluid in a small container. SOLUTION: A stirring device for stirring a fluid in a container covered with a lid includes a stirrer rotating around a rotating axis and a rotating magnetic field generating means for generating a rotating magnetic field. The stir bar has a mixture and a magnet or pedestal with a magnetic material that receives a rotating magnetic field. The outer surface of the mixture has a fluid suction port and a discharge port, and the inside of the mixture has a hole connecting the suction port and the discharge port. Configure. The mixing element has a plurality of first through holes, and a part or all of the first through holes in the mixture partially overlaps with the first through holes of adjacent mixing elements by shifting their positions. Further, the fluid is communicated with the first through hole of the adjacent mixing element so as to be able to flow, and is arranged so as to divide the mixing element into the stacking direction and the extending direction. [Selection diagram] Fig. 1

Description

  The present invention relates to an agitation device, an agitation method, a cell culture method, and a reaction promotion method for agitating a fluid accommodated in, for example, a small container.

  A small container called a dish is a flat dish-like container formed from a bottom plate and side walls arranged on the outer periphery thereof, and is covered with a lid and used for culture of microorganisms and cells, or widely for testing and research. In order to stir the fluid contained in the dish for culturing, there is a combination of shaking or a magnetic stirrer and a rod-shaped stirrer (Patent Documents 1 and 2).

JP 2007-136443 A JP 2006-150221 A

  Physiological shaking that mimics the interior of a living body is expected to promote cell differentiation and proliferation. However, normal unidirectional and monotonous shaking tends to cause the fluid in the container to overflow, and the rod-shaped stirrer does not mix well. Further, when culturing adherent cells, the rod-shaped stirrer damages the cells at the bottom of the container by the rotation of the stirrer.

  This invention is made | formed in view of the said situation, and it aims at providing the stirring apparatus and stirring method which make it possible to stir the fluid in a small container efficiently and rapidly especially. Another object of the present invention is to provide a cell culture method and a reaction promotion method that enable efficient cultivation of microorganisms and cells.

The stirring device according to the present invention comprises:
A stirring device for stirring fluid contained in a container covered with a lid,
A stirrer that stirs the fluid by rotating about a rotation axis, and a rotating magnetic field generation unit that generates a rotating magnetic field that rotates the stirrer;
The stirrer has a mixture that stirs a fluid, and a pedestal including a magnet or a magnetic body that receives a rotating magnetic field,
The outer surface of the mixture has a fluid inlet and outlet,
Inside the mixture, there are one or more holes connecting the suction port and the discharge port,
The mixture is constituted by a laminate in which a plurality of mixing elements are laminated in the rotation axis direction,
The mixing element has a plurality of first through holes serving as holes inside the mixture,
In the mixture, the mixing element is arranged so that a part or all of the first through hole partially overlaps the first through hole of the adjacent mixing element while shifting its position. It is set as the structure arrange | positioned so that a fluid may be distribute | circulated between the 1st through-holes of a mixing element so that a circulation is possible, and it divides | segments into the lamination direction and extension direction of a mixing element.

The stirring method according to the present invention comprises:
A stirring method for stirring fluid contained in a container covered with a lid using the stirring device,
The outer surface of the mixture constituting the stirrer has a fluid inlet and outlet connected by holes inside the mixture,
The stirrer is disposed on the inner flat surface of the upper part of the lid and rotated, and the stirrer sucks the fluid in the container from the suction port, causes the fluid to flow through the hole inside the stirrer, In this method, the fluid is stirred into the container.
In the stirring method, the fluid in the container is used as a cell culture solution and the cell culture solution is stirred with the stirring bar, or the fluid in the container is used as a reaction solution and the reaction solution is stirred with the stirring bar. Thus, a reaction promoting method for promoting the reaction can be obtained.

  According to the present invention, since the fluid flow in the container is disturbed by the rotation of the stirring bar, the fluid in the container can be mixed efficiently. Further, since the fluid is divided inside the mixture, the fluid can be highly mixed and the fluid inside the container can be mixed efficiently. In addition, by causing a physiologically similar agitation, it is possible to reproduce a living body and draw out a target reaction.

It is sectional drawing which shows the stirring apparatus by embodiment. It is a perspective view (Drawing 2 (a)) and a side view (Drawing 2 (b)) showing a stirring bar. It is a perspective view (Drawing 3 (a)) and a side view (Drawing 3 (b)) showing a base, and the figure (c) is a side view showing a modification of a support projection. It is a top view which shows the mixing element which comprises a mixture. It is a schematic diagram which shows the flow state of the fluid inside a mixture, The figure (a) is a plane schematic diagram which shows the flow of radial direction, The figure (b) is a cross-sectional schematic diagram which shows the flow of a lamination direction. It is a photograph which shows the stirring element of the integral thing produced with 3D printer.

Hereinafter, actual forms of the present invention will be described with reference to the accompanying drawings.
As an embodiment of the present invention, the agitation method using the agitation apparatus shown in FIG. The culture solution (fluid A) is applied to a cell culturing method or the like, or the container 6 is filled with a fluid A made of a reaction promoting solution and covered with a lid 61, and the reaction promoting solution (fluid A) is covered. The present invention can be applied to a reaction promoting method for promoting the reaction by stirring. In this case, in the cell culture method, stirring can be performed while controlling the temperature of the culture solution or the culture environment by a temperature control device (not shown), and in the reaction promoting method, the temperature control device (not shown) is used. It can stir, controlling the temperature of a reaction liquid or reaction environment. In addition, this invention is not restricted to these, It can apply to various stirring methods.
In addition, as a direction described in this specification, the “stacking direction” is synonymous with the rotation axis direction, the vertical direction, and the like of the stirrer 1 (including the mixture 2 and the pedestal 3). Is synonymous with the direction orthogonal to the stacking direction, the radial direction of the mixing element 21, the circumferential direction, and the like.

  The stirrer shown in FIG. 1 includes a stirrer 1 for stirring the fluid A and a magnetic stirrer (rotating magnetic field generating means) 4 that generates a rotating magnetic field for rotating the stirrer 1. The magnetic stirrer 4 is driven by an AC power source or the like.

  The magnetic stirrer 4 is placed on the outer flat surface 61 a of the lid 61 covering the container 6, and the stirrer 1 sucked by a magnet (not shown) attached to the motor in the magnetic stirrer 4 It arrange | positions so that the inner plane 61b may be touched. The stirring bar 1 is rotated by receiving a rotating magnetic field from the magnetic stirrer 4 and stirs a fluid A such as a sample accommodated in the container 6. 2 (a) and 2 (b), the stirrer 1 is a mixture 2 that stirs the fluid A by rotating about the rotation axis S, and a stirrer that supports the mixture 2 and receives a rotating magnetic field. And a pedestal 3 having built-in magnets 5a and 5b. The pedestal 3 may include a magnetic body instead of the stirring magnets 5a and 5b. The vertical positional relationship of the stirrer 1 will be described below with the mixture 2 at the top and the pedestal 3 at the bottom. In the usage mode of FIG. 1, the stirrer 1 is in the state of the stirrer 1 shown in FIG. The upper and lower positions are oppositely arranged, and the stirrer 1 in FIG. 1 corresponds to FIG. 2 with the lid 61 side as the lower part and the container 6 side as the upper part.

  As shown in FIGS. 3A and 3B, the pedestal 3 includes a prismatic rod-like body 31 that is placed horizontally. At both ends of the upper surface of the rod-shaped body 31, screw cylinders 33 for attaching and fixing the mixture 2 are provided. Further, stirring magnets 5 a and 5 b that receive a rotating magnetic field from the magnetic stirrer 4 are accommodated at both ends of the rod-shaped body 31, respectively. The stirring magnets 5a and 5b are arranged so that the magnetization direction is perpendicular to the longitudinal direction of the rod-shaped body 31, and the magnetic pole positions of the stirring magnets 5a and 5b are opposite to each other. Has been placed. That is, in the horizontal state of the rod-shaped body 31, for example, when the north pole of one stirring magnet 5a is disposed on the lower surface side, the south pole is disposed on the lower surface side of the other stirring magnet 5b. . As the stirring magnets 5a and 5b, cylindrical, ring, or square magnets can be used.

  The rod-shaped body 31 is provided with an annular portion 32 for stabilizing the rotation of the stirring bar 1. The annular portion 32 is connected to both ends of the rod-shaped body 31 so as to be flush with the lower surface of the rod-shaped body 31. At the center of the lower surface of the rod-shaped body 31, an arc-shaped support protrusion 34 that projects into contact with the inner flat surface 61b of the upper portion of the lid 61 is projected. Thereby, since the support protrusion 34 becomes a rotation center, the stirrer 1 is supported with little movement at the time of rotation. Therefore, the frictional resistance between the stirrer 1 and the inner flat surface 61b on the top of the lid 61 becomes very small, and the stirrer 1 can be smoothly rotated.

  Two or more arc-shaped stabilizing protrusions may be provided at equal intervals on the lower surface of the annular portion 32 (including the continuous portion of the rod-shaped body 31) at a protrusion height of the support protrusion 34 or less. In this case, even if the posture of the stirring bar 1 supported by the support protrusion 34 is inclined at the time of rotation, the stabilization protrusion can be kept in a stable posture by contacting the inner flat surface 61b of the upper portion of the lid 61. Further, as shown in FIG. 3C, the support protrusion 34 may be formed over the entire lower surface of the rod-shaped body 31.

  As shown in FIGS. 2 (a) and 2 (b), the mixture 2 is composed of a laminate in which a plurality of (in this case, four) mixing elements 21 each made of a disk are stacked. 21 is fastened to the screw cylinder portion 33 of the pedestal 3 by bolts 11 (fixing means) at two positions 180 degrees on the outer peripheral portion and fixed to the pedestal 3. Thus, the mixed body 2 in which the plurality of stacked mixing elements 21 are integrated so as to be disassembled can be easily configured and fixed to the base 3. In addition, since the plurality of mixing elements 21 are configured to be disassembled individually, the mixing elements 21 are disassembled into the respective mixing elements 21 and remain in the mixing elements 21 (a first through hole 22 and a second through hole 23 described later). It is possible to easily perform a cleaning operation such as removal of residues and foreign matters. The structure for integrating the plurality of mixing elements 21 and the structure for attaching the mixture 2 to the pedestal 3 are not limited to fixing with bolts 11, but may be a dismountable attachment method such as an uneven fitting structure. Good.

  As shown in FIG. 4, the mixture 2 is configured by alternately stacking two types of mixing elements 21 a and 21 b, and each of the two types of mixing elements 21 a and 21 b penetrates in the thickness direction. A plurality of first through holes 22 are provided. The plurality of first through holes 22 are provided along the extending surfaces extending in the extending direction of the substantially disc-shaped mixing elements 21a and 21b. A central portion of the mixing elements 21 a and 21 b has a second through hole 23 having an opening area larger than that of the first through hole 22. The 2nd through-hole 23 is formed in the substantially circular shape, and when the mixing elements 21a and 21b are laminated | stacked, it is the cylindrical hollow part to which the 2nd through-hole 23 was connected with the mixture 2. 24 is formed. The upper opening of the hollow portion 24 constitutes the fluid A suction port 20α. The central axis of the hollow portion 24 coincides with the rotation axis S of the stirring bar 1. Accordingly, the suction port 20α is disposed at a position on the rotation axis S. The mixing elements 21a and 21b may be different in the number of partition walls in the circumferential direction and the radial direction from those shown in FIG.

  The first through hole 22 is formed in a substantially rectangular shape in plan view, and is arranged concentrically with the center point of the second through hole 23 as the center. The first through holes 22 are arranged in a zigzag pattern, and the arrangement patterns of the first through holes 22 are different in the two types of mixing elements 21a and 21b. In one mixing element 21a, the first through hole 22 is closed on the inner peripheral surface of the second through hole 23 and is open on the outer peripheral surface, whereas the other mixing element 21b is in the first through hole 22. Is open on the inner peripheral surface of the second through-hole 23 and closed on the outer peripheral surface. Each of the first through holes 22 opened to the outer peripheral surface of the mixing element 21a constitutes a discharge port 20β for the fluid A. Therefore, the discharge port 20β is disposed at a position outside the rotation axis S with respect to the suction port 20α that is the upper end opening of the hollow portion 24 (for example, a radially outer position orthogonal to the rotation axis S).

  The size of the first through-hole 22 can be formed to be the same size in the same circumferential direction of the mixing elements 21a and 21b, and to increase as it goes outward in the radial direction of the mixing elements 21a and 21b. . Further, in the overlapping state of the two types of mixing elements 21a and 21b, the area of the portion where the first through holes 22 overlap each other is uniform in the circumferential direction, but may be formed non-uniformly. .

  The first through holes 22 of each of the adjacent mixing elements 21a and 21b in the mixture 2 are arranged so as to be partially displaced in the radial direction and the circumferential direction so as to overlap each other, and the stacking direction of the mixing elements 21a and 21b And it is connected in the extending direction. In other words, the partition walls between the first through holes 22 extending in the radial direction and the circumferential direction of the mixing elements 21a and 21b are arranged at different positions between the adjacent mixing elements 21a and 21b. Yes. Therefore, the inside of the mixture 2 is allowed to pass through the fluid A sequentially between the first through holes 22 of the adjacent mixing elements 21a and 21b, and is divided in each of the stacking direction and the extending direction of the mixing elements 21a and 21b. It is configured. As described above, the plurality of first through holes 22 in the mixture 2 are opened to the upper opening of the hollow portion 24 serving as the suction port 20α and the outer peripheral surface of the mixing element 21a serving as the discharge port 20β. A plurality of flow paths for connecting between the two are configured.

  The mixing elements 21a and 21b and the pedestal 3 constituting the mixture 2 are made of a resin such as polyethylene, polypropylene, and fluorine resin, but may be made of ceramic, metal, or the like. By making the mixing elements 21a and 21b and the pedestal 3 made of resin, the mixing elements 21a and 21b and the pedestal 3 can be easily and inexpensively manufactured by resin molding. When the mixing elements 21a and 21b and the pedestal 3 are made of a resin other than the fluorine resin, the surface layer may be formed of a fluorine resin by coating or the like. In this case, chemical resistance is improved.

  On the other hand, a motor having two driving magnets is disposed inside the magnetic stirrer 4 (not shown). The two drive magnets are in the same direction as the magnetization directions of the stirring magnets 5a and 5b of the base 3 of the stirrer 1, and the magnetic poles (N pole and S pole) are opposite to each other. Is arranged. The two driving magnets are rotated by the rotation of the motor to generate a rotating magnetic field.

  Next, a method for stirring the fluid A using the stirring bar 1 having the above configuration will be described. As shown in FIG. 1, the magnetic stirrer 4 is placed on the outer flat surface 61a on the top of the lid 61 that covers the container 6 containing the fluid A, and stirred so that the pedestal 3 side is in contact with the inner flat surface 61b on the top of the lid 61. Child 1 is placed. At this time, since the stirring magnets 5a and 5b accommodated in the stirrer 1 are adsorbed by the driving magnet inside the magnetic stirrer 4, the stirrer 1 is not in contact with the bottom of the container 6 and the stirrer 1 is 6 is in a floating state. Then, the motor of the magnetic stirrer 4 is driven, and the driving magnet attached to the motor is rotated. Then, the stirring magnets 5a and 5b of the pedestal 3 of the stirrer 1 are attracted to the rotating magnetic field generated by the rotating driving magnet inside the magnetic stirrer 4, and the stirrer 1 in contact with the inner flat surface 61b above the lid 61 is Rotated.

  The rotation of the stirrer 1 causes the fluid A held inside the mixture 2 to receive centrifugal force and flow toward the outer peripheral portion of the mixture 2, and the first of the mixing elements 21 a that open to the outer peripheral surface of the mixture 2. It flows out of the mixture 2 from the through hole 22. On the other hand, the fluid A in the container 6 flows into the hollow portion 24 from the suction port 20α of the mixture 2 toward the upper part of the container 6 in which the mixture 2 is disposed. The fluid A that has flowed into the hollow portion 24 receives centrifugal force due to the rotation of the stirrer 1, and flows into the mixture 2 from the first through hole 22 of the mixing element 21 b that opens on the inner peripheral surface of the hollow portion 24. The fluid A passes through the other first through hole 22 communicating with the first through hole 22 from the first through hole 22 at the inflow location, and further communicates with the other first through hole 22. It flows through the inside of the mixture 2 so as to pass through one through-hole 22, and flows out from the outlet 20β of the outer periphery of the mixture 2. The fluid A flowing out from the outlet 20β of the mixture 2 circulates in the container 6 and is taken into the mixture 2 from the suction port 20α of the mixture 2 again. In this way, the fluid A is agitated.

  In this case, as shown in FIG. 5A, the fluid A flowing through the mixture 2 passes through the plurality of first through holes 22 of the mixing elements 21a and 21b and travels from the inner peripheral portion toward the outer peripheral portion. At this time, the fluid A is divided in the extending direction of the mixing elements 21a and 21b and merges.

  In addition, since the mixture 2 includes a plurality of mixing elements 21a and 21b in the stacking direction, a plurality of fluids A are also circulated in the stacking direction of the mixing elements 21a and 21b through the first through holes 22 communicating in the stacking direction. A flow path is formed. Therefore, when the fluid A inside the mixture 2 passes through the first through hole 22, as shown in FIG. 5B, the fluid A also flows in the stacking direction of the mixing elements 21a and 21b. Are also divided in the stacking direction of the mixing elements 21a, 21b, merged and mixed.

  The mixture 2 may include one mixing element 21a, 21b in the stacking direction. Even if the mixing elements 21a and 21b are stacked one by one, the fluid A in the container 6 flows into the hollow portion 24 from the suction port 20α of the mixture 2 and receives the centrifugal force due to the rotation of the stirring bar 1, It flows into the inside of the mixture 2 from the hollow portion 24, passes through the communicating first through hole 22, and flows out from the outlet 20β of the outer periphery of the mixture 2. Even if it does in this way, it is because the fluid A is divided | segmented into the extension direction of mixing element 21a, 21b, and merges and is mixed.

  Thus, the flow of the fluid A inside the mixture 2 spreads not only in the planar or two-dimensional division and merging in the extending direction of the mixing elements 21a and 21b but also in the stacking direction of the mixing elements 21a and 21b. Dividing and merging are performed three-dimensionally. By such a three-dimensional flow, the fluid A is mixed by repeating division, merging, turbulent flow, and the like. When the mixing elements 21a and 21b are stacked one by one, planar and two-dimensional division and merging are performed, but even in this case, the fluid A is repeatedly mixed, divided, merged, and turbulent. The Further, the fluid A may only be divided inside the mixture 2. Even in this case, the fluid A that has flowed out of the mixture 2 joins and mixes inside the container 6.

  By the way, in the conventional stirrer made of a rotating body such as a rod or cloth, extremely high shear stress is generated around the stirrer, and cells and the like are cultured in a dish (flat dish 6). The cells are damaged. On the other hand, the central portion in the container 6 is a uniform and relatively slow flow, and sufficient stirring is often not obtained.

  In contrast, the stirrer 1 of the present embodiment includes a hollow portion 24 that penetrates in the center of the mixture 2 in the direction of the rotation axis S. Therefore, the fluid A at the center in the container 6 is sucked into the hollow portion 24 from the suction port 20α of the mixture 2 and rapidly mixed by being highly agitated inside the mixture 2.

  The hollow portion 24 of the mixture 2 is formed by laminating a second through hole 23 having a larger opening area than the first through hole 22, and opens sufficiently large with respect to the first through hole 22. Therefore, the inflow resistance of the fluid A to the hollow portion 24 is smaller than that of the first through hole 22 adjacent to the suction port 20α. Further, since the hollow portion 24 of the mixture 2 is located at the center of rotation of the stirrer 1, the suction port 1 does not stagnate the fluid A in the container 6 facing the hollow portion 24 due to the rotation of the stirrer 1. It can be sucked into the hollow part 24 through 20α. Then, the fluid A sucked into the hollow portion 24 receives a centrifugal force due to the rotation of the mixture 2 and passes through each first through-hole 22, thereby stacking the mixing elements 21 a and 21 b as described above. Dividing and joining are repeated in the direction and the extending direction, respectively, and the mixture is highly mixed and flows out of the mixture 2.

  As described above, according to the stirrer 1 provided with such a mixture 2, the fluid A inside the container 6 at the center of rotation can be rapidly stirred, and the entire fluid A can be efficiently stirred, and as a result, the container The time until the whole fluid A in 6 is uniformly stirred can be shortened. Moreover, since the stirrer 1 does not contact the bottom of the container 6, the cells attached to the bottom of the container 6 are not damaged. Furthermore, if an inverted microscope is used, it is also possible to take a picture while the stirring bar 1 is operating from the bottom of the container 6.

  In addition, this invention is not limited to the above embodiment, A various change is possible within the range of the summary of this invention, For example, it can deform | transform as follows.

  The mixed body 2 can be constituted by an integrated body provided with the first through hole 22 and the hollow portion 24 described above, not a laminate in which a plurality of mixing elements 21 are laminated so as to be disassembled. Such a unitary mixture 2 can be easily manufactured by, for example, a 3D printer apparatus. Further, the other mixture 2 may be a porous integrated body having continuous pores to be the first through holes 22 and the above-described hollow portion 24 provided therein. In the stirring bar 1, the pedestal 3 and the mixture 2 can be configured as a single body. Incidentally, the stirrer 1 shown in FIG. 6 is an acrylic UV curable resin stirrer 1 manufactured by a 3D printer device. The stirrer 3 is embedded with a stirring magnet 5a, 5b and bonded thereto. is there.

  Alternatively, the container 6 may be placed on the magnetic stirrer 4 and the stirring bar 1 may be disposed at the bottom of the container 6. This is because the fluid A in the container 6 is mixed by the stirrer 1 even in this way. Further, in the case of culture of floating cells, even if the stirring bar 1 is rotated at the bottom inside the container 6, it is difficult to damage the cells.

  Also with the mixture and the stirring device of the other forms described above, the fluid A inside the container 6 at the center of rotation can be rapidly stirred and the entire fluid A can be efficiently stirred as in the above-described embodiment. As a result, the time until the entire fluid A in the container 6 is uniformly stirred can be shortened.

1 Stirrer 2 Mixture 3 Pedestal 4 Magnetic Stirrer 6 Container

Claims (11)

A stirring device for stirring fluid contained in a container covered with a lid,
A stirrer that stirs the fluid by rotating about a rotation axis, and a rotating magnetic field generation unit that generates a rotating magnetic field that rotates the stirrer;
The stirrer has a mixture that stirs a fluid, and a pedestal including a magnet or a magnetic body that receives a rotating magnetic field,
The mixture is constituted by a laminate in which a plurality of mixing elements are laminated in the rotation axis direction,
The mixing element has a plurality of first through holes serving as holes inside the mixture,
In the mixture, the mixing element is arranged so that a part or all of the first through hole partially overlaps the first through hole of the adjacent mixing element while shifting its position. A stirrer configured to communicate with a first through hole of the mixing element so as to be able to flow and to be divided into a stacking direction and an extending direction of the mixing element. The rotating magnetic field generating means is disposed on the outer flat surface of the upper part of the lid,
The stirrer according to claim 1, wherein the stirrer is disposed on an inner flat surface of an upper portion of the lid so as to be rotated by the rotating magnetic field generating means. The mixing element in the mixture has a second through hole larger than the first through hole;
3. The stirring device according to claim 1, wherein the second through-hole communicates in the stacking direction in the mixture, and a hollow portion for allowing a fluid to flow into the mixture is formed.   The stirring device according to any one of claims 1 to 3, wherein the mixture in the stirrer is fixed to the pedestal while a plurality of mixing elements are fixed in a stacked state so as to be disassembled by fixing means. .   The stirrer according to any one of claims 1 to 4, wherein the mixture in the stirrer is formed of a single piece.   The stirring device according to any one of claims 1 to 5, wherein a surface layer of the mixture and the pedestal constituting the stirring bar is formed of a fluororesin. A stirring method for stirring fluid contained in a container covered with a lid using the stirring device according to any one of claims 1 to 6,
The outer surface of the mixture constituting the stirrer has a fluid inlet and outlet connected by holes inside the mixture,
The stirrer is disposed on the inner flat surface of the upper part of the lid and rotated, and the stirrer sucks the fluid in the container from the suction port, causes the fluid to flow through the hole inside the stirrer, A stirring method in which a fluid is stirred by flowing into the container. A cell culture method using the stirring method according to claim 7,
The fluid in the container is a cell culture solution,
A cell culture method in which a cell culture solution is stirred with the stirring bar. The cell culture method according to claim 8,
A cell culture method in which the cell culture solution is stirred while controlling the temperature of the cell culture solution or the culture environment. A reaction promoting method by the stirring method according to claim 7,
The fluid in the container is a reaction solution,
A reaction promoting method for promoting a reaction by stirring a reaction solution with the stirring bar. In the reaction promotion method of Claim 10,
A reaction promoting method of stirring the reaction solution while controlling the temperature of the reaction solution or the reaction environment.