Sampling device and program

Abstract

A controller for this sampling device operates an agitation element at a basic speed during a period in which a liquid circulates in a container by means of a circulation flow path (step S10). In this way, the liquid in the container is agitated by the agitation element. The controller causes a flow path switching unit to switch the flow path after a given amount of time has passed since an operation speed of the agitation element was reduced from the basic speed (step S18). In this way, the liquid circulating in the circulation flow path flows into a branch flow path.

Description

Technical Field

[0001] The present disclosure relates to a sampling device and a program for sampling a liquid in a container.

Background Art

[0002] As disclosed in Patent Document 1, there is known a device for culturing cells such as microorganisms by adjusting the dissolved oxygen concentration while stirring a culture solution in a container.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] In the device disclosed in Patent Document 1, there was a variation in the proportion of gas in the liquid (such as a culture solution) sampled by suction, and there was a possibility that accurate suction and fractionation of the solution could not be performed.

[0005] The present disclosure has been made to solve such problems, and an object thereof is to provide a technique for keeping the amount of dissolved gas in the sampled liquid constant and realizing accurate sampling of the liquid.

Means for Solving the Problems

[0006] A first aspect of this disclosure relates to a sampling device for sampling liquid in a container, comprising: a circulation mechanism that circulates the liquid in the container through a circulation channel by leading the liquid out of the container into a circulation channel and introducing the liquid from the circulation channel into the container; a stirring bar provided in the container for stirring the liquid in the container; a flow path switching unit provided in the middle of the circulation channel that can switch the flow path so that the liquid circulating in the circulation channel flows out to a branch channel for sampling; and a controller that controls the operation of the stirring bar and the flow path switching unit, wherein the controller operates the stirring bar at a basic speed during the period in which the liquid in the container is circulating through the circulation channel, thereby stirring the liquid in the container with the stirring bar, and after a given time has elapsed since the operating speed of the stirring bar was reduced from the basic speed, the controller switches the flow path to the flow path switching unit so that the liquid circulating in the circulation channel flows out to a branch channel for sampling.

[0007] A second aspect of this disclosure relates to a program executed by a computer that controls a sampling device for sampling liquid in a container. The sampling device includes a circulation mechanism that circulates the liquid in a container by leading the liquid out of the container into a circulation channel and introducing the liquid from the circulation channel into the container; a stirring bar provided inside the container for stirring the liquid in the container; and a channel switching unit provided in the middle of the circulation channel that can switch the channel so that the liquid circulating in the circulation channel flows out to a branch channel for sampling. The program is executed by a computer to cause the computer to perform the following steps: to cause the stirring bar to stir the liquid in the container by operating it at a basic speed during the period in which the liquid in the container is circulating through the circulation channel; and to cause the channel switching unit to switch the channel so that the liquid circulating in the circulation channel flows out to a branch channel for sampling after a given time has elapsed since the operating speed of the stirring bar was reduced from the basic speed. [Effects of the Invention]

[0008] According to this disclosure, variations in the amount of dissolved gas in the sampled liquid are suppressed between multiple samplings, enabling accurate sampling of the liquid. [Brief explanation of the drawing]

[0009] [Figure 1] This is a diagram showing the schematic configuration of the automated preprocessing system. [Figure 2] This is a flow path diagram showing the flow path configuration of the sampling device. [Figure 3] This is a block diagram illustrating the schematic configuration of the control device. [Figure 4] This is a perspective view of a cell culture apparatus. [Figure 5] This is a plan view of a cell culture apparatus with some parts removed. [Figure 6] This is a section cross-sectional view taken from VI-VI in Figure 5. [Figure 7] This is a section cross-sectional view taken along line VII-VII in Figure 5. [Figure 8] This diagram shows the internal structure of a cell culture device. [Figure 9] This is a diagram showing a stirring bar. [Figure 10] This diagram shows the stirring bar removed from the shaft. [Figure 11] This is a flowchart of the process performed by the sampling device 1 to sample the culture medium from the cell culture device 100 into a test tube 14. [Figure 12] This figure shows an example of the result of the liquid introduction amount following the procedure in Figure 11. [Figure 13] This figure shows an example of the results for the amount of liquid introduced, following the comparative example. [Modes for carrying out the invention]

[0010] This embodiment will be described in detail with reference to the drawings. Note that identical or corresponding parts in the drawings are denoted by the same reference numerals, and their descriptions will not be repeated in principle.

[0011] <Schematic Configuration of Automatic Pretreatment System> FIG. 1 is a block diagram showing a schematic configuration of an automatic pretreatment system 10. The automatic pretreatment system 10 is a device for automatically performing pretreatment on an analysis target. In the present embodiment, the analysis target is, for example, cultured cells, and more specifically, bacterial cells.

[0012] The automatic pretreatment system 10 includes a sampling device 1 and a pretreatment device 2. After pretreatment is performed by the automatic pretreatment system 10, metabolites of the cells are extracted from the cells. The extracted metabolites are supplied to a liquid chromatograph mass spectrometer 3. The liquid chromatograph mass spectrometer 3 is merely an example of an analysis device for analyzing an analysis target. It is also possible to analyze the analysis target using other analysis devices.

[0013] The sampling device 1 is a device for sampling liquid from a container (culture container). For example, microorganisms and plant cells are cultured in a culture solution containing a culture medium in a container called a bioreactor. In the bioreactor, for example, a stirring member rotated using a magnetic force, an oxygen concentration sensor for detecting the concentration of dissolved oxygen, and the like are provided. By adjusting the dissolved oxygen concentration while stirring the culture solution containing the culture medium and cells in the bioreactor, cells are cultured in the sampling device 1. A detailed description of the bioreactor that functions as a cell culture device will be described later.

[0014] The pretreatment device 2 performs pretreatment on the cells contained in the culture solution (culture sample) sampled from the bioreactor. In the sampling device 1, the culture solution containing cells is accommodated in a test tube as a container (sampling container). The pretreatment device 2 includes a centrifugation mechanism 4, a liquid removal mechanism 5, a reagent supply mechanism 6, a stirring mechanism 7, an extraction mechanism 8, and the like. These mechanisms sequentially perform pretreatment on the cells contained in the culture solution in the test tube.

[0015] The centrifugal separation mechanism 4 applies a centrifugal force to the culture solution in the test tube. As a result, the culture solution in the test tube is separated into a solid component that sinks to the bottom of the test tube with the solid-liquid interface as the boundary and a liquid component that floats on top of the solid component. The solid component is a culture, for example, cultured cells. The liquid component that floats on top of the solid component is the supernatant separated from the culture solution.

[0016] The liquid removal mechanism 5 sucks out the supernatant from the test tube. As a result, the liquid in the test tube is removed, and cells remain in the test tube. The reagent supply mechanism 6 supplies a reagent for extracting metabolites in the cells to the cells in the test tube. As a result, a mixed solution of cells and the reagent is generated in the test tube. The stirring mechanism 7 stirs the mixed solution. By stirring the mixed solution, a suspension in which metabolites are extracted from the cells is obtained.

[0017] The extraction mechanism 8 extracts a part of the suspension as an extract. The extract is supplied to the liquid chromatography mass spectrometer 3.

[0018] <Schematic configuration of the sampling device> FIG. 2 is a flow path diagram showing the flow path configuration of the sampling device 1. In the sampling device 1, a culture solution containing cells in a cell culture device 100 called a bioreactor is sampled. Inside the cell culture device 100, a magnetic stirrer 111 is provided as a stirring member rotated using magnetic force.

[0019] The cell culture device 100 is held by a holding portion 12 provided inside the sampling device 1. In the present embodiment, three cell culture devices 100 can be held by one holding portion 12, and a plurality (for example, four) of such holding portions 12 are provided. The holding portion 12 may be provided in a configuration where only one is provided. The holding portion 12 may also be configured to hold two or less or four or more cell culture devices 100.

[0020] The cell culture apparatus 100 can perform cell culture while heated by a heater (not shown) provided in the holding section 12. A motor 13 for rotating magnets (not shown) is connected to the holding section 12. By rotating this motor 13, the magnets are rotated, and the resulting magnetic force can rotate the stirring bars 111 inside each cell culture apparatus 100.

[0021] The sampling device 1 can perform cell culture by controlling the temperature of the culture medium in the cell culture apparatus 100 and stirring the culture medium with the stirring bar 111. The sampling device 1 can sample the culture medium containing the cultured cells into a test tube 14 at any desired timing.

[0022] The sampling device 1 is equipped with a culture medium sampling mechanism 20 for sampling the culture medium into a test tube 14, and a reagent sampling mechanism 30 for sampling the reagent into a test tube 14. The test tube 14 contains a mixture of the culture medium and the reagent, is sealed with a cap (not shown), and then transported to the pretreatment device 2.

[0023] The culture medium sampling mechanism 20 is equipped with a pump 21 and a number of valves 22 and 23. Each valve 23 has, for example, one pair of common ports and five pairs (10 in total) of selective ports. The flow path can be switched by arbitrarily selecting one pair of selective ports and connecting it to one pair of common ports.

[0024] The pump 21 and valve 22 are installed in a flow path 41 that connects a pair of common ports. The valve 22 constitutes a flow path switching unit (first flow path switching unit) for switching whether or not to guide the liquid in the flow path 41 to a branch flow path 42 that branches off from the flow path 41. In other words, the valve 22 can switch between a state in which liquid flows between the pair of common ports via the flow path 41, or a state in which the liquid in the flow path 41 is guided to the branch flow path 42.

[0025] Of the five pairs of selection ports, one pair is connected to an outlet 43 and an inlet 44, respectively, which communicate with one cell culture device 100. The outlet 43 is a channel for draining the culture medium from within the cell culture device 100. The inlet 44, on the other hand, is a channel for introducing the culture medium, which is drained from the cell culture device 100 via the outlet 43 and circulated via the channel 41, back into the cell culture device 100. Another pair of selection ports is connected to an outlet 45 and an inlet 46, respectively, which communicate with another cell culture device 100. Yet another pair of selection ports is connected to an outlet 47 and an inlet 48, respectively, which communicate with yet another cell culture device 100.

[0026] In the sampling device 1, any of the outlet passages 43, 45, or 47 and their corresponding inlet passages 44, 46, or 48 are connected via the flow path 41, and the pump 21 is driven in this state, thereby circulating the culture medium within each cell culture device 100. In other words, the flow path 41, each outlet passage 43, 45, or 47, and each inlet passage 44, 46, or 48 constitute a circulation flow path (first circulation flow path) for circulating the culture medium within each cell culture device 100.

[0027] The pump 21 discharges the culture medium from each cell culture device 100 into the first circulation channel and introduces the culture medium into each cell culture device 100 from the first circulation channel, thereby constituting a circulation mechanism (first circulation mechanism) that circulates the culture medium within each cell culture device 100 via the first circulation channel.

[0028] Each outlet 43, 45, and 47 has its tip immersed in the culture medium within the corresponding cell culture device 100. On the other hand, each inlet 44, 46, and 48 has its tip positioned above and spaced apart from the culture medium within the corresponding cell culture device 100. The culture medium discharged from the cell culture device 100 via each outlet 43, 45, and 47 and circulating through the channel 41 is introduced into the cell culture device 100 by falling from the tips of each inlet 44, 46, and 48.

[0029] In the sampling device 1, at least the portion of the flow path 41 connecting a pair of common ports that is equipped with a pump 21 is made of a flexible tube. The pump 21 is, for example, a tubing pump, which can pump the liquid inside the tube by deforming (compressing and relaxing) the flexible tube.

[0030] By switching the valve 22, which serves as a first channel switching unit located in the middle of channel 41, the culture medium circulating into each cell culture device 100 via channel 41 can be discharged into the branched channel 42. At this time, the tip of the branched channel 42 is located inside the test tube 14, and the culture medium is sampled into the test tube 14 via the branched channel 42.

[0031] Of the two pairs of selection ports other than the three pairs to which each of the outlets 43, 45, 47 and each of the inlets 44, 46, 48 are connected, one pair of selection ports is connected to the cleaning solution tank 26 and the waste liquid tank 27, respectively. The remaining pair of selection ports is connected to the filter 25 and the waste liquid tank 27, respectively. The cleaning solution tank 26 contains a cleaning solution for cleaning the flow path of the culture medium.

[0032] After sampling the culture medium from one of the cell culture devices 100 into a test tube 14, the valve 23 is switched to connect the washing solution tank 26 and the waste liquid tank 27 to the flow path 41. With the pump 21 then driven, the washing solution in the washing solution tank 26 is discharged into the waste liquid tank 27 via the flow path 41. This allows the flow path 41 and the valve 22, etc., to be cleaned with the washing solution.

[0033] After cleaning with the cleaning solution, the valve 23 is switched to connect the filter 25 and the waste liquid tank 27 to the flow path 41. When the pump 21 is then driven, air is introduced into the flow path 41 via the filter 25 and discharged into the waste liquid tank 27 along with any remaining moisture in the flow path 41. This removes moisture from the flow path 41 and the valve 22, etc.

[0034] The reagent sampling mechanism 30 is equipped with a pump 31 and a number of valves 32, 33. Each valve 33 has, for example, one common port and multiple selective ports, and the flow path can be switched by arbitrarily selecting one of the selective ports and connecting it to the common port.

[0035] Pump 31 and valve 32 are installed in a flow path 49 that communicates with the reagent tank 34 at both ends. The reagent tank 34 contains reagents to be mixed with the culture medium sampled in the test tube 14. Flow path 49 constitutes a circulation flow path (second circulation flow path) for circulating the reagents in the reagent tank 34. Pump 31 discharges reagents from the reagent tank 34 into the second circulation flow path and introduces reagents into the reagent tank 34 from the second circulation flow path, thereby constituting a circulation mechanism (second circulation mechanism) for circulating the reagents in the reagent tank 34 via the second circulation flow path.

[0036] In the sampling device 1, at least the portion of the flow path 49, which is connected at both ends to the reagent tank 34, that contains the pump 31 is made of a flexible tube. The pump 31 is, for example, a tubing pump, and by deforming (compressing and relaxing) the flexible tube, it can deliver the reagent inside the tube.

[0037] Valve 32 constitutes a flow path switching unit (second flow path switching unit) for switching whether or not to direct the liquid in flow path 49 to branch flow path 50 which branches off from flow path 49. In other words, valve 32 can switch between a state in which the reagent in reagent tank 34 is circulated via flow path 49, or a state in which the reagent in flow path 49 is directed to branch flow path 50.

[0038] In this way, by switching the valve 32, which acts as a second flow path switching unit located in the middle of the flow path 49, the reagent circulating into the reagent tank 34 via the flow path 49 can be made to flow out into the branched flow path 50. The branched flow path 50 is connected to the common port of the valve 33, and one of the selectable ports of the valve 33 is connected to the inside of the test tube 14. Therefore, by connecting the selectable port connected to the inside of the test tube 14 to the common port, the reagent flowing out from the flow path 49 into the branched flow path 50 can be sampled into the test tube 14.

[0039] <Outline configuration of the control device> Figure 3 is a block diagram showing the schematic configuration of the control device 60. The sampling device 1 is equipped with the control device 60. The control device 60 includes, for example, a CPU (Central Processing Unit) 61 and a memory 62. The memory 62 is composed of, for example, ROM (Read Only Memory) and RAM (Random Access Memory), and can store various data in addition to control programs. The CPU 61 can control the operation of the motor 13, pumps 21, 31 and valves 22, 23, 32, 33, etc., by executing the control programs stored in the memory 62.

[0040] The control device 60 can circulate the culture medium in any of the cell culture devices 100 by driving the pump 21 at a constant liquid delivery rate while any of the outlet passages 43, 45, 47 and the corresponding inlet passages 44, 46, 48 are connected via the flow path 41. Based on the control program, the control device 60 switches the valve 22 for a predetermined time, connecting the flow path 41 and the branched flow path 42, thereby allowing the culture medium in the flow path 41 to be sampled into a test tube 14.

[0041] The control device 60 can control the amount of culture medium sampled by controlling the time it takes to switch the flow path with the valve 22. In other words, if the pumping speed of the pump 21 is known in advance, the desired amount of culture medium can be accurately sampled into the test tube 14 by adjusting the time it takes to connect the flow path 41 and the branched flow path 42.

[0042] The control device 60 can circulate the reagents in the reagent tank 34 by driving the pump 31 at a constant liquid delivery rate while the flow path 49 is connected from one end to the other. Based on the control program, the control device 60 switches valve 32 for a predetermined time to connect the flow path 49 and the branch flow path 50, and also switches valve 33 to connect the branch flow path 50 to the test tube 14, thereby allowing the reagents in the flow path 49 to be sampled into the test tube 14.

[0043] The control device 60 can control the amount of reagent sampled by controlling the time it takes to switch the flow path with the valve 32. In other words, if the liquid delivery speed of the pump 31 is known in advance, the desired amount of reagent can be accurately sampled into the test tube 14 by adjusting the time it takes to connect the flow path 49 and the branched flow path 50.

[0044] <Configuration of cell culture apparatus> Figures 4 to 10 illustrate the structure of the cell culture apparatus 100. Figure 4 is a perspective view of the cell culture apparatus 100, Figure 5 is a plan view of the cell culture apparatus 100 with some parts removed, Figure 6 is a section cross-section of Figure 5 from VI-VI, Figure 7 is a section cross-section of Figure 5 from VII-VII, Figure 8 shows the internal structure of the cell culture apparatus 100, Figure 9 shows the stirring bar 111, and Figure 10 shows the state with the stirring bar 111 removed from the shaft 110.

[0045] As shown in Figure 4, the cell culture apparatus 100 includes a container 101, a lid 102, a DO (Dissolved Oxygen) sensor 103 connected to the lid 102, a pH sensor 104, a cap 105, and a shaft 110.

[0046] Container 101 is a transparent container into which a culture medium containing microorganisms, plant cells, etc. is placed. The lid 102 is for sealing container 101 and has various components attached to it. The DO sensor 103 is a sensor for measuring the dissolved oxygen concentration in the cell culture apparatus 100. The pH sensor 104 is a sensor for measuring the hydrogen ion concentration in the culture medium. The cap 105 is a lid with an opening that protrudes from the top of the cell culture apparatus 100. The shaft 110 is a component that serves as the shaft of the stirring bar 111, allowing the stirring bar 111 attached to its tip to rotate.

[0047] Figures 5 to 7 show the DO sensor 103 and pH sensor 104 with some components removed. In the plan view of Figure 5, five pipes connected to a flexible tube are provided between the DO sensor 103 and the pH sensor 104. The five pipes include an oxygen intake pipe 121, an oxygen exhaust pipe 122, a sample addition pipe 123, an intake pipe 124, and an exhaust pipe 125.

[0048] The oxygen intake pipe 121 is a pipe for supplying oxygen to the culture medium and extends to the stirring bar 111 located below the container 101, as shown in Figures 6 and 7. The oxygen exhaust pipe 122 is a pipe for exhausting excess oxygen from the cell culture apparatus 100. The sample addition pipe 123 is a pipe for adding samples as needed. The intake pipe 124 is connected to one of the introduction passages 44, 46, or 48 described above and is a pipe for introducing the culture medium into the cell culture apparatus 100. The discharge pipe 125 is connected to one of the outflow passages 43, 45, or 47 described above and is a pipe for discharging the culture medium from the cell culture apparatus 100 to the outside of the cell culture apparatus 100.

[0049] The lengths of the five pipes are described below. The oxygen exhaust pipe 122 is the shortest of the five pipes and extends to a position where it overlaps with the lid 102 in the vertical direction (downward direction in the plane of the paper). The sample addition pipe 123 and the suction pipe 124 are approximately the same length and are longer than the oxygen exhaust pipe 122, extending vertically to the center of the container 101.

[0050] The oxygen intake pipe 121 is longer than the sample addition pipe 123 and the suction pipe 124, and extends to a position where it overlaps with the stirring bar 111 in the vertical direction. The discharge pipe 125 is longer than the oxygen intake pipe 121, and extends to a position below the stirring bar 111 in the vertical direction.

[0051] The five pipes are fixed together with the shaft portion 110 to the base portion 110c on the upper surface of the cover portion 102. Three baffle plates 110b extend downward from the base portion 110c, as shown in Figure 8. The ends of the baffle plates 110b are fixed to the annular portion 110a. Of the five pipes, the oxygen intake pipe 121 and the exhaust pipe 125 are fixed to the annular portion 110a.

[0052] The baffle plate 110b is a component for creating turbulence, generating both vertical and horizontal flows in addition to the horizontal flow caused by the rotation of the agitator 111. The agitator 111 has a magnet placed inside the magnet section 111d. As shown in Figures 6 to 9, the end 121a of the oxygen intake pipe 121 is located in a position that overlaps with the agitator 111 in the vertical direction, and the end 125a of the discharge pipe 125 is located below the agitator 111 in the vertical direction.

[0053] Thus, since the end 121a of the oxygen intake pipe 121 is positioned to overlap with the stirring bar 111, and the end 125a of the discharge pipe 125 is positioned below the stirring bar 111, the culture medium can be discharged outside the container 101 at a position where it is less affected by oxygen bubbling caused by oxygen supply, even if the culture medium is stirred by the stirring bar 111. Therefore, the cell culture apparatus 100 is designed to allow the following: Cultivation This method allows for accurate aspiration and separation of samples while maintaining a constant amount of dissolved gas in the nutrient solution.

[0054] Next, the agitator 111 will be described in detail. As shown in Figure 10, the agitator 111 includes a main body 111c, a bearing portion 111a, and a locking portion 111b. The main body 111c is formed in a cylindrical shape with a hole 111f in the center, and rotating blade portions 111e are formed every 90°. The agitator 111 includes two magnetic portions 111d that protrude from the main body 111c. The magnetic portions 111d are covered with the same material as the main body 111c.

[0055] The main body 111c of the stirring bar 111 is made of polyetheretherketone. Polyetheretherketone is commonly referred to as PEEK, and will therefore be referred to as PEEK below. The material covering the rotor blade section 111e and the magnet section 111d, which are integrally formed with the main body 111c, is also made of PEEK. In contrast, the bearing section 111a and the locking section 111b are made of polyacetal (POM).

[0056] Polyacetal and PEAK are both resins, but they differ in their properties. Polyacetal, due to the mixture of amorphous and crystalline regions, is used as an engineering plastic with excellent strength, elastic modulus, and impact resistance. Because of its excellent sliding properties, polyacetal is also used in bearing components. In contrast, PEAK is classified as a super engineering plastic with the highest performance. Among super engineering plastics, PEAK is known as a highly reliable resin with particularly excellent heat resistance and chemical resistance.

[0057] The peaks constituting the rotor blade section 111e have higher strength and wear resistance than the polyacetal constituting the bearing section 111a. The polyacetal constituting the bearing section 111a is self-lubricating and has a particularly low coefficient of friction with metal. The interior of the shaft section 110 is formed of metal such as stainless steel (e.g., SUS316). The polyacetal constituting the bearing section 111a is suitable as a bearing member because it has higher sliding properties against metal than the peaks constituting the rotor blade section 111e.

[0058] As shown in Figures 9 and 10, the agitator 111 has a bearing portion 111a inserted into a hole 111f of the main body 111c, which is composed of peaks. The bearing portion 111a is rotatably positioned on a metal shaft inside the shaft portion 110, and its bottom surface is fixed by a locking portion 111b.

[0059] The stirring bar 111 has a bearing portion 111a made of polyacetal which has self-lubricating properties and a low coefficient of friction with metal. Therefore, even if the bearing portion 111a slides against the shaft portion 110 for a long time, no shavings are produced. The stirring bar 111 has a rotating blade portion 111e which is made of peaks, so even if it rotates for a long time , a stirring bar and No debris is generated between the oxygen intake pipe 121 and the exhaust pipe 125, which are in contact with each other. This allows for stable analysis of microbial metabolites.

[0060] In this embodiment in particular, as shown in Figures 6 to 8, the end of the baffle plate 110b is fixed to the annular portion 110a, and the agitator 111 rotates at a position below the annular portion 110a. As a result, the baffle plate 110b does not slide against the part of the agitator 111 that covers the rotating blade portion 111e or the magnetic portion 111d, thus preventing the generation of debris. This allows for stable analysis of microbial metabolites.

[0061] <Sampling process flow> Figure 11 is a flowchart of the process performed by the sampling device 1 to sample the culture medium from the cell culture apparatus 100 into a test tube 14. In one implementation example, the sampling device 1 performs the process shown in Figure 11 by the CPU 61 of the control device 60 executing a given program.

[0062] The control device 60 is an example of a controller that controls the operation of the agitator (agitator 111) and the flow path switching unit (valve 22). The control device 60 controls the operation of the agitator 111 by controlling the operation of the motor 13.

[0063] The program may be stored in memory 62. In this case, memory 62 is an example of a recording medium for non-temporarily storing the program. The program may also be stored in a recording medium accessible by the CPU 61 and detachable from the control device 60. In this case, the recording medium is an example of a recording medium for non-temporarily storing the program.

[0064] The process shown in Figure 11 is performed for each test tube 14. For the sake of clarity, the following description will focus on sampling from the leftmost of the three cell culture devices 100 shown in Figure 2. In this description, the channel 41, together with the outlet channel 43 and the inlet channel 44, constitutes the first circulation channel. The following description will refer to Figure 11 to explain the process flow.

[0065] In step S10, the control device 60 causes the cell culture apparatus 100 to perform basic operations. These basic operations include circulating the culture medium in the first circulation channel and circulating reagents in the second circulation channel. Circulating the culture medium in the first circulation channel includes rotating the motor 13 to rotate the stirring bar 111, thereby agitating the culture medium inside the cell culture apparatus 100.

[0066] In step S12, the control device 60 determines whether or not it is time to sample the culture medium into the test tube 14. The control device 60 repeats the determination in step S12 until it determines that the timing has arrived (NO in step S12). When the control device 60 determines that the timing has arrived (YES in step S12), it proceeds to step S14.

[0067] In step S14, the control device 60 stops the rotation of the motor 13, thereby stopping the rotation of the stirring bar 111. This stops the stirring of the culture medium inside the cell culture device 100.

[0068] In step S16, the control device 60 determines whether a given time has elapsed since the stirring was stopped in step S14. The control device 60 continues the control in step S16 until it determines that the given time has elapsed (NO in step S16), and when it determines that the given time has elapsed (YES in step S16), it proceeds to step S18.

[0069] In step S18, the control device 60 causes the valve 22 to switch the flow path so that the liquid in the flow path 41 is directed to the branch flow path 42.

[0070] In step S20, the control device 60, after a specific time has elapsed since the valve 22 was switched in step S18, instructs the valve 22 to switch the flow path so that the liquid in the flow path 41 is directed to the introduction path 44. The specific time in step S20 refers to the time corresponding to sampling a given amount.

[0071] In step S22, the control device 60 restarts the rotation of the motor 13, thereby restarting the agitation of the culture medium in the cell culture apparatus 100. After that, the control device 60 terminates the process shown in Figure 11.

[0072] In the process described above with reference to Figure 11, in step S18, the flow path is switched by valve 22 for sampling into test tube 14. Note that stirring in the cell culture apparatus 100 is stopped a given time before the flow path is switched in step S18 (step S14). That is, after stirring is stopped in step S14, and after a given time has elapsed, the flow path is switched in step S18 and sampling begins.

[0073] In the cell culture apparatus 100, sampling is performed after stirring is stopped for a "given time," which suppresses variations in the amount of dissolved gas in the sampled culture medium across multiple samplings. If the "given time" is too short, the variation in the amount of dissolved gas for each sample will not be sufficiently suppressed. On the other hand, if the "given time" is too long, it is conceivable that uneven culture conditions will occur in the sampled culture medium with each sample. In this sense, in one implementation example, the "given time" may be set between 2 and 10 minutes. In another implementation example, the "given time" may be set between 3 and 7 minutes.

[0074] Furthermore, if variations in the amount of dissolved gas for each sample are suppressed, stirring does not need to be stopped completely. That is, instead of stopping the rotation of the stirring bar 111 in step S14, the control device 60 may reduce the rotation speed of the stirring bar 111 from the rotation speed in the basic operation in step S10. The control device 60 reduces the rotation speed of the stirring bar 111 by reducing the rotation speed of the motor 13. The rotation speed of the stirring bar 111 in step S10 is also called the "basic speed" and is the speed set for the circulation of the culture medium.

[0075] In one implementation example, the control device 60 reduces the rotational speed of the agitator 111 to about 1 / 10 of the basic speed in step S14. In this case, after sampling is completed and the flow path is switched in step S20, the control device 60 returns the rotational speed of the agitator 111 to the rotational speed at the time in step S10.

[0076] <Uniformity of the amount introduced into the test tube> Figure 12 shows an example of the results of the amount of liquid introduced by introducing the liquid according to the procedure in Figure 11. Figure 13 shows an example of the results of the amount of liquid introduced by introducing the liquid according to the comparative example. The results in Figures 12 and 13 are for when pure water is used as the liquid introduced from the cell culture apparatus 100 to the test tube 14. In the graphs in Figures 12 and 13, the vertical axis represents the weight of the liquid introduced into the test tube 14.

[0077] The results shown in Figure 12 are those obtained when stirring by the stirring bar 111 is stopped before a given time has elapsed since the liquid was introduced from the cell culture apparatus 100 into the test tube 14, as explained with reference to Figure 11. On the other hand, the results shown in Figure 13 are those obtained when stirring by the stirring bar 111 is continued before and after the liquid was introduced from the cell culture apparatus 100 into the test tube 14.

[0078] In both Figure 12 and Figure 13, the range of liquid introduction volumes (maximum and minimum) for each of the 10 groups (A1 to E2) during the 6 liquid introductions is shown. Note that the target liquid introduction volume differs for each group. The target introduction volume for groups A1 and A2 is 2 mL, for groups B1 and B2 it is 1 mL, for groups C1 and C2 it is 0.5 mL, for groups D1 and D2 it is 0.2 mL, and for groups E1 and E2 it is 0.1 mL.

[0079] The results in Figure 13 show that in all groups, the difference between the maximum and minimum weights of the liquid introduced into test tube 14 is greater than the results in Figure 12.

[0080] For example, in Figure 13, in group A1, the maximum value was 1.65g, the minimum value was 1.50g, and the difference between the maximum and minimum values ​​was 0.15g. Since the median was 1.575g, the difference of 0.15g between the maximum and minimum values ​​was a relatively high value, approximately 10% of the median of 1.575g.

[0081] Furthermore, in Figure 13, in group B2, the maximum value was 0.90g, the minimum value was 0.60g, and the difference between the maximum and minimum values ​​was 0.30g. Since the median was 0.75g, the difference between the maximum and minimum values, 0.30g, was a relatively high value, representing approximately 43% of the median of 0.75g.

[0082] On the other hand, the results in Figure 12 show that in all groups, the difference between the maximum and minimum weights of the liquid introduced into test tube 14 is small. In other words, the results in Figure 12 show that in all groups, there is almost no variation in the weight of the liquid introduced into test tube 14. That is, as explained with reference to Figure 11, if stirring by the stirring bar 111 is stopped a given time before the liquid is introduced from the cell culture apparatus 100 into test tube 14, the variation in the proportion of gas in the aspirated solution is suppressed, and thereby the accuracy of the aspirated solution can be controlled. It should be noted that this effect can be expected not only when stirring by the stirring bar 111 is completely stopped, but also when the rotational speed for stirring by the stirring bar 111 is reduced.

[0083] [Aspect] Those skilled in the art will understand that the above-described exemplary embodiments are specific examples of the following embodiments.

[0084] (Clause 1) A sampling device according to one embodiment is a sampling device for sampling liquid in a container, comprising: a circulation mechanism that circulates the liquid in the container through a circulation channel by leading the liquid out of the container into a circulation channel and introducing the liquid from the circulation channel into the container; a stirring bar provided in the container for stirring the liquid in the container; a channel switching unit provided in the middle of the circulation channel that can switch the channel so as to allow the liquid circulating in the circulation channel to flow out to a branch channel for sampling; and a controller that controls the operation of the stirring bar and the channel switching unit, wherein the controller operates the stirring bar at a basic speed during the period in which the liquid in the container is circulating through the circulation channel, thereby causing the stirring bar to stir the liquid in the container, and after a given time has elapsed since the operating speed of the stirring bar was reduced from the basic speed, the controller switches the channel to the channel switching unit so as to allow the liquid circulating in the circulation channel to flow out to a branch channel for sampling.

[0085] According to the sampling apparatus described in paragraph 1, before the flow path is switched for sampling, the operating speed of the agitator that stirs the liquid in the container is reduced for a given time. This reduces variations in the amount of dissolved gas in the sampled liquid between multiple samplings, thereby enabling accurate sampling of the liquid.

[0086] (Clause 2) In the sampling apparatus described in paragraph 1, reducing the operating speed of the stirring bar from the basic speed may include stopping the stirring bar.

[0087] According to the sampling apparatus described in paragraph 2, variations in the amount of dissolved gas in the sampled liquid can be more reliably suppressed between multiple samplings.

[0088] (3) In the sampling apparatus described in paragraph 1 or 2, the length of the given time may be 2 minutes to 10 minutes.

[0089] According to the sampling device described in paragraph 3, the situation in which variations in the amount of dissolved gas for each sample are not sufficiently suppressed due to the "given time" being too short is avoided. Furthermore, the situation in which uneven culture occurs in the culture medium sampled for each sample due to the "given time" being too long is avoided.

[0090] (Clause 4) In the sampling apparatus described in paragraph 1 or 2, the length of the given time may be 3 to 7 minutes.

[0091] According to the sampling device described in Section 4, the situation in which variations in dissolved gas amounts for each sample are not sufficiently suppressed due to an excessively short "given time" can be more reliably avoided. Furthermore, the situation in which uneven culture conditions occur in the sampled culture medium for each sample due to an excessively long "given time" can be more reliably avoided.

[0092] (Clause 5) A program according to one embodiment may be a program executed by a computer that controls a sampling device for sampling liquid in a container. The sampling device may include a circulation mechanism that circulates the liquid in the container through the circulation channel by leading the liquid out of the container into a circulation channel and introducing the liquid from the circulation channel into the container; a stirrer provided in the container for stirring the liquid in the container; and a channel switching unit provided in the middle of the circulation channel that can switch the channel so that the liquid circulating in the circulation channel flows out to a branch channel for sampling. The program may be executed by the computer to cause the computer to perform the steps of: operating the stirrer at a basic speed during the period in which the liquid in the container is circulating through the circulation channel, thereby causing the stirrer to stir the liquid in the container; and, after a given time has elapsed since the operating speed of the stirrer was reduced from the basic speed, causing the channel switching unit to switch the channel so that the liquid circulating in the circulation channel flows out to a branch channel for sampling.

[0093] According to the program described in Section 5, before the flow path is switched for sampling, the operating speed of the agitator that stirs the liquid in the container is reduced for a given time. This reduces the variation in the amount of dissolved gas in the sampled liquid between multiple samplings, thereby enabling accurate sampling of the liquid.

[0094] (Clause 6) In the program described in paragraph 5, reducing the operating speed of the stirring bar from the basic speed may include stopping the stirring bar.

[0095] According to the program described in Section 6, variations in the amount of dissolved gas in the sampled liquid can be more reliably suppressed between multiple samplings.

[0096] (Clause 7) In the program described in paragraph 5 or 6, the length of the given time may be between 2 minutes and 10 minutes.

[0097] According to the program described in Section 7, the situation in which variations in dissolved gas amounts for each sample are not sufficiently suppressed due to a "given time" being too short is avoided. Furthermore, the situation in which uneven culture conditions occur in the sampled culture medium for each sample due to a "given time" being too long is avoided.

[0098] (Clause 8) In the program described in paragraph 5 or 6, the length of the given time may be between 3 and 7 minutes.

[0099] According to the program described in Section 8, the situation in which variations in dissolved gas amounts for each sample are not sufficiently suppressed due to a "given time" being too short is more reliably avoided. Furthermore, the situation in which uneven culture conditions occur in the sampled culture medium for each sample due to a "given time" being too long is more reliably avoided.

[0100] The embodiments disclosed herein should be considered in all respects to be illustrative and not restrictive. The scope of this disclosure is indicated by the claims rather than by the description of the embodiments above, and all modifications within the meaning and scope of the claims are intended to be included. [Explanation of symbols]

[0101] 1 Sampling device, 2 Pretreatment device, 3 Liquid chromatograph mass spectrometer, 4 Centrifugal separation mechanism, 5 Liquid removal mechanism, 6 Reagent supply mechanism, 7 Stirring mechanism, 8 Extraction mechanism, 10 Pretreatment system, 12 Holding part, 13 Motor, 14 Test tube, 20 Culture medium sampling mechanism, 21, 31 Pump, 22, 23, 32, 33 Valve, 25 Filter, 26 Washing solution tank, 27 Waste liquid tank, 30 Reagent sampling mechanism, 34 Reagent tank, 41, 42, 49, 50 Flow path, 43, 45, 47 Outlet path, 44, 46, 48 Inlet path, 60 Control device, 61 CPU, 62 Memory, 100 Cell culture device, 101 Container, 102 Lid, 103 DO sensor, 104 pH sensor, 105 Cap, 110 Shaft, 110a Ring part, 110b Baffle plate, 110c base section, 111 stirring bar, 111a bearing section, 111b locking section, 111c main body, 111d magnet section, 111e rotor section, 121 oxygen intake piping, 121a, 125a end sections, 122 oxygen exhaust piping, 123 sample addition piping, 124 intake piping, 125 discharge piping.

Claims

1. A sampling device for sampling liquid inside a container, A circulation mechanism that circulates the liquid inside the container by introducing liquid from the container into a circulation channel and introducing liquid from the circulation channel into the container, A stirring bar is provided inside the container for stirring the liquid inside the container, A flow path switching unit is provided in the middle of the aforementioned circulation channel and is capable of switching the flow path so as to allow the liquid circulating in the circulation channel to flow out to a branch channel for sampling. The system includes a controller that controls the operation of the stirring bar and the flow path switching unit, The aforementioned controller, During the period in which the liquid in the container is circulated through the aforementioned circulation channel, the agitator is operated at a basic speed to agitate the liquid in the container. In accordance with the timing of each of the multiple samplings of the liquid circulating in the aforementioned circulation channel, the operating speed of the agitator is reduced from the basic speed. After reducing the operating speed of the agitator from the basic speed and a predetermined time has elapsed, the flow path switching unit is switched so that the liquid circulating in the circulation path flows out to the branched flow path for sampling. A sampling device in which the length of the given time is between 2 and 10 minutes.

2. The sampling apparatus according to claim 1, wherein reducing the operating speed of the stirring bar from the basic speed includes stopping the stirring bar.

3. The sampling apparatus according to claim 1, wherein the length of the given time is between 3 and 7 minutes.

4. A program executed by a computer that controls a sampling device for sampling liquid in a container, The sampling device is A circulation mechanism that circulates the liquid inside the container by introducing liquid from the container into a circulation channel and introducing liquid from the circulation channel into the container, A stirring bar is provided inside the container for stirring the liquid inside the container, The circulation channel includes a channel switching unit provided in the middle of the circulation channel, which is capable of switching the channel so as to allow the liquid circulating in the circulation channel to flow out to a branch channel for sampling, The program is executed by the computer, thereby causing the computer to: During the period in which the liquid in the container is circulated through the circulation channel, the stirring bar is operated at a basic speed to stir the liquid in the container; The steps include reducing the operating speed of the agitator from the basic speed in accordance with the timing of each of the multiple samplings of the liquid circulating in the circulation channel, After reducing the operating speed of the agitator from the basic speed and a predetermined time has elapsed, the flow path is switched at the flow path switching unit so that the liquid circulating in the circulation flow path is discharged into the branch flow path for sampling. A program in which the length of time given above is between 2 minutes and 10 minutes.

5. The program according to claim 4, wherein reducing the operating speed of the stirring bar from the basic speed includes stopping the stirring bar.

6. The program according to claim 4, wherein the length of the given time is between 3 and 7 minutes.

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