Multi-chamber gas concentration difference generation system

Abstract

We want to be able to generate a desired gas concentration difference between multiple containment chambers containing cell samples, etc., with relatively high precision using gas control in two separate spaces, and to enable long-term experiments. [Solution] In the internal gas circulation container 3, the containment chamber, which is partitioned by openings 23 and 7, is closed at the bottom by a chamber slide 13 and at the top by a glass plate 29, forming a sealed chamber 62. Adjacent sealed chambers 62, 62 are in ventilated communication via through holes 27 to gas adjustment chambers 32. In the gas adjustment chamber 32, the internal gas circulating inside the internal gas circulation container 3 and the external gas are separated by a porous membrane sheet 31, and gas exchange occurs if there is a difference in gas concentration. An arbitrary difference in the concentration of O2 gas in the internal gas is generated between adjacent sealed chambers 62, 62 separated by the gas adjustment chamber 32. The gas concentration of the internal gas in each sealed chamber 62 is maintained with higher precision than when adjusting using oxygen absorbers, etc. Moreover, it can accommodate long-term experiments.

Description

Technical Field

[0001] The present invention relates to a multi-chamber gas concentration difference generation system, and particularly to a multi-chamber gas concentration difference generation system suitable for generating a gas concentration difference in a plurality of storage chambers in a culture device provided with the plurality of storage chambers.

Background Art

[0002] When considering optimizing the culture environment for specimens such as cells, tests are conducted by changing the culture conditions for a plurality of specimens, and the gas environment is also a major factor in the culture conditions.

[0003] Normally, as described in Patent Document 1, since the gas inside the container and the gas outside the container are naturally and gradually exchanging gas, in order to maintain the gas concentration inside the container with high precision, the gas controlled from the gas supply device is supplied and constantly replaced, and the plurality of specimens placed inside the container are all in the same gas environment. If different gas environments are to be created for each specimen, a plurality of containers are provided and the gas concentration in each container is controlled. Since the gas concentration in each container is maintained at an arbitrary gas concentration with high precision, an arbitrary gas concentration difference can be generated with high precision between the plurality of containers, but as the number of containers increases, the control becomes more complicated and the cost also increases.

[0004] On the other hand, if a plurality of specimens are separately placed in inner containers configured using an oxygen absorber or the like and then collectively housed in a large container to which gas is supplied, a plurality of gas environments can be created in one large container, but it is difficult to accurately control the gas concentration difference between the inner containers. Moreover, since the oxygen absorber or the like is a consumable and deteriorates relatively early and cannot absorb gas, it cannot cope with long-term experiments.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

[0006] This invention addresses the above-mentioned conventional problems and aims to provide a novel and useful multi-chamber gas concentration difference generation system that can generate arbitrary gas concentration differences in multiple containment chambers with relatively high accuracy and can also be used for long-term experiments. [Means for solving the problem]

[0007] The present invention was made to solve the above problems, and is a multi-chamber gas concentration difference generation system characterized by a sealed containment chamber in which a specimen holder for cells or the like can be set with an open airflow above, a communication means for connecting a plurality of the containment chambers in a row in a breathable manner, a gas inlet and a gas outlet provided on one end and the other end of the plurality of containment chambers in the row direction, the communication means comprising a gas adjustment chamber that adjusts the gas concentration depending on the gas concentration difference inside and outside separated by an open-cell type porous membrane, and a ventilation passage connecting the gas adjustment chamber and the containment chamber, and the internal gas flowing out from the upstream containment chamber flows into the downstream containment chamber via the gas adjustment chamber.

[0008] Preferably, the internal gas separated by the porous membrane flows in through the gas inlet at a controlled gas concentration, and the external gas fills the outside of the porous membrane at a controlled gas concentration. Preferably, the porous membrane is replaceable.

[0009] Preferably, a gas adjustment chamber for a communication means is formed in a space partitioned by a longitudinal partition wall of the internal gas circulation container, a ventilation passage is formed by a through-hole penetrating the longitudinal partition wall, and the gas adjustment chamber is removably closed with a porous membrane sheet. Preferably, the internal gas circulation container is configured to accommodate the specimen holder so that it can be observed under a microscope. Preferably, once the specimen holder is set, the containment chamber is sealed, allowing ventilation only through the ventilation channel.

[0010] Preferably, an inner gas circulation container is housed in an outer container, and the gap between the outer container and the inner gas circulation container is filled with outside gas. In one example, the outer container is designed to be placed on the microscope stage. [Effects of the Invention]

[0011] According to the multi-chamber gas concentration difference generation system of the present invention, by managing the gas in two spaces, it is possible to generate any desired gas concentration difference between multiple containment chambers with relatively high accuracy, and it can also be used for long-term experiments. [Brief explanation of the drawing]

[0012] [Figure 1] This is a perspective view of an internal gas flow container that constitutes a multi-chamber gas concentration difference generation system according to an embodiment of the present invention. [Figure 2] Figure 1 is an exploded perspective view of the internal gas flow container. [Figure 3] Figure 2 is a perspective view of the flow channel section. [Figure 4] Figure 2 is a decomposed perspective view from a different direction. [Figure 5] This is a cross-sectional view of Figure 1. [Figure 6] Figure 1 is a perspective view of the outer container containing the inner gas flow container. [Figure 7] Figure 6 is an exploded perspective view. [Figure 8] This is a cross-sectional view of Figure 6. [Figure 9] Figure 1 is an explanatory diagram of the gas exchange operation using the porous membrane sheet provided. [Figure 10] Figure 6 shows a schematic diagram illustrating the configuration of a multi-chamber gas concentration difference generation system. [Figure 11] Figure 10 is an explanatory diagram illustrating the generation pattern of the gas concentration difference. [Modes for carrying out the invention]

[0013] The multi-chamber gas concentration difference generation system 1 according to the embodiment of the present invention will be described with reference to the drawings. In FIG. 1, the vertical, front-rear, left-right directions of the inner gas flow container 3 are defined by arrows, and the following description will be based on this direction. FIGS. 1 to 5 mainly show the inner gas flow container 3. This inner gas flow container 3 is in the shape of a rectangular box in plan view, and the bottom is composed of a base frame 5 in the shape of a rectangle in plan view. In this base frame 5, three rectangular openings 7, 7, 7 are arranged in parallel such that the long-side edges face each other, and the longitudinal direction of each opening 7 is formed in a direction orthogonal to the longitudinal direction of the base frame 5. Further, the openings 7, 7, 7 are closer to one side in the longitudinal direction of the base frame 5, that is, the left side. An inner flange 7a is formed on the lower edge side of this opening 7.

[0014] On the upper surface of the base frame 5, an inner portion along a shape similar to a rectangular contour and grooves are formed at the boundaries between adjacent openings 7 and 7 and are connected in a cross shape, and an attachment seal 9 is attached thereto. Further, holes are formed at the four corners of the base frame 5, and dish screws 11, 11, 11, 11 are inserted from the lower surface side. Note that a step is formed on the outer edge of the upper surface of the base frame 5, which is one step lower.

[0015] A chamber slide 13 is set in this opening 7. The edge of the slide 13a of the chamber slide 13 is placed on the inner flange 7a and set. The chamber slide 13 is a kind of specimen mounting tool, and the upper surface of the slide 13a is the mounting surface for specimens such as cells. The upper opening of this chamber slide 13 is closed by a covering lid. Since the covering lid is only placed by its own weight, it is in a ventilation state even when closed by the covering lid.

[0016] The two chamber slides 13, 13 are closed by transparent covers 15, 15. The remaining chamber slide 13 is closed by a cover 17. The cover 17 is almost the same overall shape as the cover 15, but the cover frame 17a forms part of the side and top surfaces, and a glass plate 17b is fitted into the remaining part of the top surface. An insertion opening 17c for a temperature sensor (more precisely, a tip detection part) 19 is formed in the top surface formed by this cover frame 17a. This insertion opening 17c consists of a groove extending inward from the edge of the top surface and a through hole that penetrates the top surface downward from the tip of the groove. The base end of the L-shaped temperature sensor 19 is fitted into the groove, and the curved end is inserted through the through hole and hangs downward. A cover 17 is placed over the leftmost chamber slide 13 of the three chamber slides 13, 13, 13 arranged in a row, and the insertion port 17c is located on the upper left side.

[0017] Reference numeral 21 indicates a integrally molded frame-shaped channel compartment. This channel compartment 21 has a rectangular outer contour in plan view, and the outer edge of its lower surface is stepped down. It is mounted on the base frame 5, and this step engages with a step on the base frame 5 side, so that the outer surface is almost flush with the base frame 5 side. The flow channel section 21 is partitioned by a longitudinal septum 21a, and inside, in plan view, there are large, wide rectangular openings 23, 23, 23 and small, narrow, oval openings 25, 25. In the engagement state described above, the edges of the openings 23 are almost flush with the edges of the openings 7 of the base frame 5, and the three openings 23, 23, 23 and the three openings 7, 7, 7 are connected vertically to each other separately. The chamber slide 13, which is set in the opening 7, is surrounded by the rim of this opening 23.

[0018] The openings 25, 25 have bottoms and are positioned on the front and rear sides of the flow channel section 21, respectively, with their longitudinal directions aligned with the longitudinal direction of the flow channel section 21. They are perpendicular to the longitudinal direction of the openings 23, 23, 23. Furthermore, the longitudinal dimension of the opening 25 is approximately the same as the sum of the short dimensions of the adjacent openings 23 and the boundary portion, with the rear opening 25 straddling the left opening 23 and the middle opening 23, with its center at the boundary portion, and the front opening 25 straddling the middle opening 23 and the right opening 23, with its center at the boundary portion.

[0019] Reference numeral 27 indicates a through hole. This through hole 27 penetrates the longitudinal septum 21a separating opening 23 and opening 25, connecting opening 23 and opening 25. The through hole 27 extends straight through the flesh portion of the longitudinal septum 21a, and its cross-section perpendicular to the direction of penetration is circular. On the side of opening 25, through holes 27, 27 are located at both ends in the longitudinal direction, and each of these through holes 27, 27 communicates with the adjacent openings 23, 23.

[0020] The eye-shaped section formed by the edges of the three openings 23, 23, 23 is lower than the rest of the structure, forming a fitted frame with a stepped bottom and stepped sides. By fitting the glass plate 29 onto this stepped bottom, the openings 23, 23, 23 are closed independently. In other words, adjacent openings 23 and 23 are in a state where ventilation is possible through the through-hole 27 and opening 25.

[0021] Furthermore, a similar fitting frame is formed around the opening 25, and a porous membrane sheet 31 is fitted onto the stepped bottom surface of this frame, separating the inside from the outside and creating a gas adjustment chamber 32. The porous membrane of the porous membrane sheet 31 is of the open-cell type and is a film that does not allow water droplets of about 100 to 2,000 μm in size to pass through, but allows water vapor (=moisture) and air of about 0.0004 μm in size to pass through. Conventionally, it has been used in filtration filters, heat insulation materials, thermal insulation materials, gas valves for ECUs and headlights, protective paper for cables, diaper back sheets, and breathable packaging materials for desiccants, etc. In this embodiment, as an example, one made of Teflon (registered trademark) is used.

[0022] While there are no particular limitations on pore size or porosity (%), if the air permeability (measured using the Oken-type air permeability tester compliant with JIS P8117, which is the time required for a specified volume of air to pass through per unit area and unit pressure difference, expressed in seconds per 100 mL) becomes too low, gas will easily escape from the porous membrane, leading to faster gas replacement and a rapid change in gas concentration. Therefore, although it is also influenced by other factors, a value of around 100 (sec / 100 mL) is considered necessary. If the air permeability is high, there is no problem even if the pore size or porosity is large. If there is a difference in gas concentration between the inside and outside of the gas adjustment chamber 32, gas will pass through the porous membrane from the area of ​​higher concentration to the area of ​​lower concentration, meaning that gas exchange occurs between the inside and outside, and the gas concentration inside is adjusted depending on the gas concentration outside. For example, as shown in Figure 9, if there is a difference in O2 concentration across the porous membrane, O2 gas will pass from the area of ​​higher concentration to the area of ​​lower concentration.

[0023] Parameters that affect the amount of gas exchange include the air permeability of the porous membrane sheet 31 and the concentration of the outside gas separated by the porous membrane sheet 31, as well as the length of the porous membrane sheet 31 that constitutes the lid of the gas adjustment chamber 32 extending in the direction of gas flow, and the cross-sectional size of the gas adjustment chamber 32 perpendicular to the direction of gas flow. The larger the gas adjustment chamber 32 is set to be, the longer the residence time of the internal gas in the gas adjustment chamber 32 will be, and the amount of gas exchange will increase. However, adjusting the amount of gas exchange would require changes to the hardware design, so it is not practical to use it as a parameter. Therefore, it is envisioned that the air permeability of the porous membrane sheet 31 and the concentration of the external gas will be used as parameters.

[0024] The flow channel section 21 is partitioned by a longitudinal septum 21a, and also has a wider, oval-shaped, bottomed opening 33 than the opening 25. This opening 33 is positioned to the right of the right-side opening 23, aligned longitudinally, and communicates with the right-side opening 23 via a through-hole 27. The opening 33 has a step formed in the same way as the opening 25, and a water receiving seal 35 is installed therein. The opening 33 is closed by a flat plate-shaped cover 37. This cover 37 has an insertion hole 37a for the pipe 39 that penetrates through the flesh portion. A ring seal 37b is fitted into this insertion hole 37a.

[0025] The L-shaped end of the pipe 39 is inserted through the insertion hole 37a, and the end extends below the lower surface of the lid 37. This end is airtightly fitted into the ring seal 37b that is attached to the insertion hole 37a. Furthermore, a pair of opposing through holes are formed in the lid 37, and when the lids 37 are stacked, a pair of screw holes are also formed on the upper surface of the flow channel section 21 opposite the pair of through holes, and knurled screws 41, 41 are inserted from above and screwed into the screw holes, and the space between the lid 37 and the opening 33 is airtightly sealed by the water receiving seal 35.

[0026] Through holes are formed coaxially in the flow channel compartment 21, communicating with the holes in the four corners of the base frame 5, and a spacer 43 with a threaded hole is fitted into these through holes. A countersunk screw 11 is screwed into the spacer 43, integrating the flow channel compartment 21 with the base frame 5, and the space between the opening 23 and the opening 7 is airtightly sealed by the attachment seal 9. Furthermore, the lower surfaces of the opposing short edges of the opening 23 are raised by one step, and a container fixing block 45 is attached to this stepped surface. By tightening, the edge of the slide 13a is clamped between the container fixing block 45 and the inner flange 7a, thereby fixing the chamber slide 13. Furthermore, a sensor fixing block 61 is fitted into the part of the temperature sensor 19 that is above the groove, and the temperature sensor 19 is pressed and fixed in place from above.

[0027] Reference numeral 47 indicates a flat cover. When viewed from above, with the lid 37 fitted into the recessed portion on the right side, the cover 47 has an outer contour that is almost identical to that of the flow channel compartment 21. The cover 47 has openings 49, 49, 49 and 51, 51 formed on its inside, with opening 49 being approximately the same size as opening 23 and opening 51 being approximately the same size as opening 25. When the plate surface of the cover 47 is superimposed on the flow channel section 21 with its outer contours aligned on three sides, the opening 49 faces the opening 23, and the opening 51 faces the opening 25, with their upper and lower edges aligned.

[0028] Through holes are formed in the four corners of the cover 47, and when the parts are stacked as described above, they are coaxially opposed to the screw holes of the spacer 43. A knurled screw 57 is inserted from above and screwed into the screw hole. In addition, through holes are formed in four places on the cover 47, and when the parts are stacked as described above, they are coaxially opposed to the screw holes formed in the flow path section 21. A bolt 59 is inserted from above and screwed into the screw hole. These fasteners create an airtight seal between the lower surface of the glass plate 29 and the upper edges of the openings 23, 23, and 23.

[0029] The internal gas circulation container 3 has translucent sections made of glass plates 17b, 29, etc., so that the specimen placed on the chamber slide 13 can be observed under a microscope. Furthermore, the drawing also shows through-holes penetrating the side surface of the flow channel section 21. These are a result of the manufacturing process and will be blocked off with pins or the like during testing.

[0030] In the internal gas circulation container 3 configured as described above, the containment chamber, which is partitioned by openings 23 and 7, is closed at the bottom by a set chamber slide 13 and at the top by a glass plate 29, forming a sealed containment chamber, i.e., a sealed chamber 62 that is connected to the outside only by through holes 27 and insertion openings 17c, which serve as ventilation passages. The sealed chambers 62A, 62B, and 62C are arranged in parallel from right to left, and adjacent sealed chambers 62, 62 are ventilated and communicate with each other via through holes 27, gas adjustment chambers 32, and through holes 27, which serve as communication means. Therefore, the sealed chambers 62A, 62B, and 62C are ventilated and communicate in a line via the communication means.

[0031] The installation method is simple: countersunk screws 11 and spacers 43 are attached to the base frame 5, and the chamber slides 13, 13, 13 on which the specimen is placed are set with the covers 15, 15, 17 placed over them. The porous membrane sheet 31 is then aligned with the openings 25, 25 on the flow channel compartment 21, and the glass plate 29 is aligned with the openings 23, 23, 23. The cover 47 is then placed on the flow channel compartment 21 and secured with bolts 59, and knurled screws 57 are inserted. The opening 33 is then closed with the lid 37. Finally, the flow channel compartment 21 is placed on the base frame 5, and the knurled screws 57 are screwed in and tightened with spacers 43 to complete the installation. Furthermore, if the disposable porous membrane sheet 31 becomes dirty, it can be easily replaced by removing the cover 47, which is bolted to the flow channel section 21.

[0032] As shown in Figures 6 to 8, this internal gas circulation container 3 is housed in an outer container 63. This outer container 63 is a type that can be observed under a microscope and is placed on a microscope stage, and is a rectangular box shape in plan view. The container body 65 consists of a bottom section and side sections, with a large rectangular opening 67 formed in the bottom section in a manner similar to its outer contour. A partition wall rises along the rim of the opening 67 on the inside, and a water tank 69 is formed in the space between the side section and the partition wall. A heater is installed on the bottom side of this water tank 69, and the water inside the water tank 69 evaporates due to the heating of the heater, thereby achieving the desired humidity level inside. The tip of the pipe 71 penetrates the side section and extends into the interior. The container body 65 is closed with a flat lid 73. The inside of this lid 73 is a translucent area and is coated with a transparent conductive film that primarily serves as a heater to warm the inside of the outer container 63. Reference numeral 75 indicates a temperature sensor (more precisely, a tip detection unit). This temperature sensor 75 is inserted into the outer container 63.

[0033] The internal gas circulation container 3 is set in the well attachment 77 and then housed in the container body 65. The well attachment 77 is a rectangular frame in plan view, with leaf springs 79 attached to the outer surfaces of each of its four side sections, and handles 81 attached to two corresponding side sections. When the well attachment 77 is lowered with its orientation aligned so that the bottom surface of the internal gas flow container 3 rests on the inner flange provided on the bottom side, the well attachment 77 becomes fitted onto the internal gas flow container 3 and is locked in that position by the leaf springs 79. Then, when the well attachment 77 is placed on the rim of the opening 67 of the container body 65, the internal gas circulation container 3 is housed inside the container body 65 together with the well attachment 77. The lid 73 is closed and locked to the container body 65 by being screwed in at multiple points.

[0034] As described above, the inner gas flow container 3 is housed in the outer container 63 in a manner that allows for microscopic observation, and concentration-controlled gases flow into the inner gas flow container 3 and the outer container 63, respectively. To distinguish between the two gases, the gas flowing into the inner gas flow container 3 is referred to as the inner gas, and the gas flowing into the outer container 63 is referred to as the outer gas.

[0035] As shown in Figure 10, a multi-chamber gas concentration difference generation system 1 is configured. The internal gas is pressurized, so as indicated by the arrow, the flow direction is unidirectional. The internal gas, whose concentration is controlled by the gas mixing device, first flows through pipe 39 into the right-hand sealed chamber 62A where the chamber slide 13 is housed. Since there is ventilation between the chamber slide 13 and the covers 15 and 17, the internal gas also flows into the chamber slide 13. The internal gas in this sealed chamber 62A flows through the through-hole 27 into the front gas adjustment chamber 32, moves longitudinally within the gas adjustment chamber 32, and then flows into the intermediate sealed chamber 62B, where it also flows into the ventilated chamber slide 13. Subsequently, the internal gas flows into the sealed chamber 62C and chamber slide 13 on the left side via the rear gas adjustment chamber 32, and finally exits through the gap between the inlet 17c and the temperature sensor 19 as the gas outlet.

[0036] Regarding the external gas, the external gas, whose concentration is controlled by the gas mixing device, is pressurized and flows into the outer container 63 through the pipe 71, which serves as the gas inlet. This external gas fills the outer container 63, and any excess flows out from the boundary between the container body 65 and the lid 73. The gas concentrations of the internal and external gases are independently controlled by a gas mixing device. In this state, as the internal gas moves within the gas adjustment chamber 32, it is separated from the external gas by the porous membrane sheet 31, allowing gas to pass through, i.e., enabling gas exchange. Furthermore, the porous membrane sheets 31, 31 that make up the two gas adjustment chambers 32, 32 are constructed with the same air permeability.

[0037] The internal gas is composed of a mixture of O2 and CO2 gases, and the external gas is also composed of a mixture of O2 and CO2 gases. However, as shown in Figure 9, for example, if the concentration of O2 gas in the internal gas is lower than the concentration of O2 gas in the external gas, the O2 gas in the external gas will pass through the porous membrane and enter the internal gas, increasing the concentration of O2 gas in the internal gas. Therefore, the concentration of O2 gas in the internal gas after it flows out is higher than the concentration of O2 gas in the internal gas before it flows into the gas adjustment chamber 32. As a result, the concentration of O2 gas in the internal gas within the sealed chamber 62 increases in the following order: right sealed chamber 62A (and chamber slide 13) < intermediate sealed chamber 62B (and chamber slide 13) < left sealed chamber 62C (and chamber slide 13).

[0038] Conversely, for example, if the concentration of O2 gas in the internal gas is higher than the concentration of O2 gas in the external gas, the O2 gas in the internal gas will pass through the porous membrane and enter the external gas, resulting in a decrease in the concentration of O2 gas in the internal gas. Therefore, the concentration of O2 gas in the internal gas after it has flowed out is lower than the concentration of O2 gas in the internal gas before it flows into the gas adjustment chamber 32. On the other hand, if the concentration of O2 gas in the internal gas and the concentration of O2 gas in the external gas are the same, no movement of O2 gas occurs across the porous membrane, and therefore the concentration of O2 gas in the internal gas does not change.

[0039] There are two gas adjustment chambers 32, and the higher the air permeability of each porous membrane sheet 31, the more gas exchange is promoted. Also, the higher the gas concentration difference between the outside gas and the inside gas, the more gas exchange is promoted. By adjusting the parameters based on these gas exchange trends, an arbitrary concentration difference in the O2 gas within the sealed chambers 62A, 62B, and 62C is generated through gas exchange. In other words, a concentration difference is generated inside the chamber slides 13, 13, and 13 housed in each of the sealed chambers 62A, 62B, and 62C.

[0040] Figure 11 shows the pattern of gas concentration difference generation. To distinguish the chamber slides 13, they are numbered No. 1, No. 2, and No. 3 from the upstream side. The numbers in the figure represent the O2 gas concentration, expressed as unitless integers for ease of comparison. The gas flows into the outer container 63 at a certain flow rate, and is immediately replaced by fresh outside gas within the outer container 63, so the gas concentration hardly changes. In pattern (A), the O2 gas concentration of the external gas is low at "2", while the O2 gas concentration of the internal gas is high at "8", but it decreases as you move from chamber slide No. 1 to No. 3. In pattern (B), the O2 gas concentration in the external gas is high at "8" and the O2 gas concentration in the internal gas is low at "2", but it increases as you move from chamber slide No. 1 to No. 3. In pattern (C), the O2 gas concentrations of the internal and external gases become the same midway through the process.

[0041] The multi-chamber gas concentration difference generation system 1 is configured in this way, and it is possible to generate a gas concentration difference between the chamber slides 13, 13, and 13, thereby simultaneously creating multiple culture conditions with different gas concentrations. Furthermore, the gas concentration within each chamber slide 13 can also be brought to a desired value by adjusting the gas concentration of the external gas and the permeability of the porous membrane sheet 31. The gas concentration can be adjusted with greater precision than when using oxygen absorbers, and the gas exchange life of the porous membrane sheet 31 is longer than that of oxygen absorbers, making it suitable for long-term experiments.

[0042] Although embodiments of the present invention have been described in detail above, the specific configuration is not limited to these embodiments, and any design changes or other modifications that do not depart from the spirit of the invention are also included in the invention. For example, if a coordinated relationship is ensured between the sealed chamber 62 and the gas adjustment chamber 32, the number of sealed chambers 62 can be increased. Furthermore, if the gas adjustment chamber 32 is sealed with a sealed lid instead of being closed with the porous membrane sheet 31, the gas concentration inside the sealed chambers 62, 62 before and after the chamber will remain unchanged and the same.

[0043] Chamber slide 13 is used for processing multiple samples and is commonly used, but the sample placement surface is not limited to this and may be made up of dishes or plates. The outer container 63 may also be made up of a CO2 incubator. [Explanation of Symbols]

[0044] 1…Multi-chamber gas concentration difference generation system 3...Internal gas circulation container 5...Base frame 7…Opening 7a…Inner flange 9... Attachment seal 11... Countersunk screw 13... Chamber slide 13a... Slide 15... Covering lid 17... Covering lid 17a...Lid frame 17b...Glass plate 17c…Socket 19…Temperature sensor 21...Flow channel compartment 21a...Longitudinal septum 23...Opening 25...Opening 27...Through hole 29...Glass plate 31…Porous membrane sheet 32…Gas adjustment chamber 33...Opening 35...Water receiving seal 37...Lid body 37a...Insertion hole 37b... Ring seal 39... Pipe 41...Knurled screw 43...Spacer 45...Container fixing block 47...Cover 49...Opening 51...Opening 57...Knurled screw 59...Bolt 61...Sensor fixing block 62...Sealed chamber 63...Outer container 65...Container body 67...Opening 69...Aquarium 71... Pipe 73... Lid 75...Temperature sensor 77...Well attachment 79...Leaf spring 81...Handle

Claims

1. A sealed containment chamber capable of holding specimens such as cells with an open top, a means for connecting multiple containment chambers in a row to allow ventilation, and a gas inlet and gas outlet provided at one end and the other end of the row of the multiple containment chambers. The aforementioned communication means consists of a gas adjustment chamber that adjusts the gas concentration depending on the difference in gas concentration between the inside and outside separated by a porous membrane of the open-cell type, and a ventilation passage connecting the gas adjustment chamber and the containment chamber. A multi-chamber gas concentration difference generation system characterized in that internal gas flowing out from the upstream containment chamber flows into the downstream containment chamber via the gas adjustment chamber.

2. In the multi-chamber gas concentration difference generation system described in claim 1, A multi-chamber gas concentration difference generation system characterized in that an inner gas separated by a porous membrane flows in from a gas inlet at a controlled gas concentration, and an outer gas fills the area outside the porous membrane at a controlled gas concentration.

3. In the multi-chamber gas concentration difference generation system described in claim 2, A multi-chamber gas concentration difference generation system characterized by a replaceable porous membrane.

4. In a multi-chamber gas concentration difference generation system according to any one of claims 1 to 3, A multi-chamber gas concentration difference generation system characterized in that a containment chamber and a gas adjustment chamber communicating with the internal gas circulation container are formed in a space partitioned by a longitudinal partition wall, a ventilation passage is formed by a through-hole penetrating the longitudinal partition wall, and the gas adjustment chamber is removably closed with a porous membrane sheet.

5. In the multi-chamber gas concentration difference generation system described in claim 4, A multi-chamber gas concentration difference generation system characterized by an internal gas flow container that is configured to accommodate a specimen holder in a way that allows for microscopic observation.

6. In the multi-chamber gas concentration difference generation system described in claim 5, A multi-chamber gas concentration difference generation system characterized in that, once the specimen placement device is set, the containment chamber is sealed, allowing ventilation only through the ventilation channel.

7. In the multi-chamber gas concentration difference generation system described in claim 6, A multi-chamber gas concentration difference generation system characterized in that an inner gas flow container is housed in an outer container, and the gap between the outer container and the inner gas flow container is filled with outer gas.

8. In the multi-chamber gas concentration difference generation system described in claim 7, A multi-chamber gas concentration difference generation system characterized by an outer container designed to be installed on the microscope stage.

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