JP2023044158A - Culture vessel lid and culture vessel - Google Patents

Abstract

To effectively suppress dew condensation that hinders observation, in a lid applied to a culture vessel used for culturing cells and the like.SOLUTION: The culture vessel lid according to the present invention is a culture vessel lid that covers the upper part of a culture vessel body that has an upper opening and a storage space that can store a liquid, comprising: two flat plate portions each made of transparent resin and having a planar size corresponding to the planar size of the culture vessel body; and a connecting portion that connects the peripheral edges of the two flat plate portions to face each other with a predetermined gap between the two flat plate portions and hermetically seals the gap space between the two flat plate portions against the external space. Since the conditions to be satisfied by thicknesses t1 and t2 of the two flat plate portions and a gap g therebetween are defined, it is possible to construct a lid that can suppress the occurrence of dew condensation for a certain period of time.SELECTED DRAWING: Figure 4

Description

The present invention relates to a lid that can be attached to a culture vessel for holding and culturing a biological sample such as cells together with a liquid.

In the technical fields of medicine and biochemistry, cells, microorganisms, tissues, etc. (hereinafter referred to as "cells, etc.") are cultured in a liquid injected into a container for the purpose of various experiments, such as microscopic observation of cells. done. For example, a culture vessel called a microplate, a well plate, a microtiter plate, or the like has a plurality of recesses called wells, each of which opens upward and is capable of holding a liquid in an internal storage space, formed on a plate-like substrate. are arranged in a matrix on the upper surface of the In addition, as a culture vessel used for the same purpose, there is, for example, a so-called flat-bottomed shallow dish.

A cycle of culture and observation (including imaging) using such a culture vessel is generally as follows. In other words, cells to be cultured are seeded in a culture vessel filled with a medium liquid to prepare a raw sample, and the culture vessel is held in an incubator maintained at a constant temperature and humidity environment. Cultivation is performed. Then, the culture vessel is taken out from the incubator at the required timing and used for observation and imaging. After that, the culture vessel is returned to the incubator. Such storage and observation in the incubator are repeated as necessary.

When the culture vessel is taken in and out of such an incubator, it is required to suppress changes in the temperature and humidity environment of the biological sample, contamination by contamination sources, and the like. For example, Patent Document 1 discloses a technique for solving this problem. In this technique, an incubator and an imaging device are integrated. Specifically, in the technique described in Patent Literature 1, both an imaging device and an incubator box are installed in a light-shielding box, and a culture vessel is housed in the incubator box for culturing. Illumination light is applied to the culture vessel through a transparent window provided in the upper part of the incubator box, and imaging is performed using an imaging optical system arranged below the culture vessel. Patent Document 1 also describes dew condensation prevention means for a transparent window.

JP 2007-166982 A

In the conventional technology described above, one set of incubator and imaging device is required for one culture vessel. On the other hand, at research sites where this type of culture is performed, it is common to accommodate a plurality of culture vessels in an incubator and culture them simultaneously. The above prior art does not correspond to such a situation, and the scale of the entire apparatus becomes large. In addition to this, devices that have a temperature control function in the culture vessel itself and devices for warming the culture vessel have been commercialized, but they require large-scale equipment and consume a lot of power. It becomes a problem.

As a more realistic and simple method in terms of housing in an incubator, it is conceivable to cover the main body of the culture vessel with a lid body provided with anti-condensation measures, for example. In this case, there is a problem that dew condensation occurs on the inside of the lid and interferes with observation. Especially when the lid is taken out from an incubator whose temperature is higher than room temperature, dew condensation occurs on the inside of the lid that has been cooled by the outside air. Cheap. It is conceivable to apply a hydrophilic chemical substance to the inside of the lid to prevent water droplets from forming due to dew condensation. This is not preferable from the viewpoint of contamination prevention.

For these reasons, it is desired to establish a technique capable of imparting an effective anti-condensation effect to the lid that covers the main body of the culture vessel, but such a technique has not yet been put to practical use.

SUMMARY OF THE INVENTION The present invention has been made in view of the above problems, and provides a technique capable of effectively suppressing dew condensation, which hinders observation, in a lid applied to a culture vessel used for culturing cells and the like. With the goal.

One aspect of the present invention is a culture vessel cover covering an upper portion of a culture vessel main body having an upper opening and a storage space capable of storing a liquid, wherein each cover is made of a transparent resin to achieve the above object. Two flat plate portions having a planar size corresponding to the planar size of the culture vessel main body and peripheral edge portions of the two flat plate portions are connected to each other, and the two flat plate portions are opposed to each other with a predetermined gap therebetween. and a connecting portion for air-tightly sealing the gap space between the two flat plate portions from the external space.

Here, of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm] is the thickness of the second flat plate portion, t2 [mm] is the thickness of the second flat plate portion, and g [mm] is the size of the gap (where t1, t2, and g are all 0.5 or more, and t1+t2+g≦6). At this time, the first flat plate portion and the second flat plate portion are made of polystyrene resin or polypropylene resin, and a predetermined dimensional relationship described later is established between t1, t2 and g.

The lid body for a culture vessel (hereinafter simply referred to as the "lid body") has a multi-layer structure having first and second flat plate portions and a gap provided therebetween, thereby improving the heat insulating performance of the lid body. can be improved. This is considered to contribute to enhancing the effect of preventing dew condensation on the lid removed from the culture environment at a temperature higher than room temperature. However, it is not enough to simply have a multi-layered structure, and although details will be described later, the range of dimensions that each part can take in order to prevent the occurrence of dew condensation is limited. Specifically, the thickness of the first flat plate portion, the thickness of the second flat plate portion, and the size of the gap must each satisfy predetermined conditions in order to suppress the occurrence of dew condensation for a certain period of time.

The inventors of the present application conducted a detailed study of the dew condensation prevention effect of such a lid body with a multilayer structure, and found that dew condensation was prevented for a certain period of time that can be said to be sufficient for observing the observation target in the culture vessel. I have found the conditions under which it is possible. As will be described later, by combining a culture vessel main body that holds an observation object such as cells together with a liquid and a lid made under the conditions proposed in this specification, dew condensation that hinders observation can be effectively eliminated. can be effectively suppressed.

As described above, according to the present invention, the cover applied to a culture vessel used for culturing cells or the like has a multi-layer structure in which two flat plate portions face each other with a gap provided therebetween. By specifying the dimensions, it is possible to construct a culture vessel that can effectively suppress dew condensation that hinders observation for a certain period of time.

1 is a diagram showing the overall structure of an example of a culture vessel according to the present invention; FIG. FIG. 4 is a diagram showing the overall structure of another example of the culture vessel according to the present invention; FIG. 4 is a cross-sectional view showing the structure of a lid; FIG. 4 is a cross-sectional view showing a configuration example of a lid; It is a figure explaining simulation conditions. FIG. 10 is a first diagram showing the size range of the lid obtained from the simulation results; FIG. 11 is a second diagram showing the size range of the lid obtained from the simulation results; FIG. 11 is a third diagram showing the dimensional range of the lid obtained from the simulation results; FIG. 10 is a fourth diagram showing the dimensional range of the lid obtained from the simulation results;

FIG. 1 is a diagram showing the overall structure of an example of a culture vessel according to the present invention. This culture vessel 10 includes a well plate 11 as a vessel body and a lid 12 . The well plate 11 has a substantially cylindrical shape (or a tapered truncated cone shape whose horizontal cross-sectional area gradually decreases toward the bottom surface) with an open top surface and a closed bottom surface on a plate-shaped base material made of, for example, a transparent resin. It has a structure in which wells W are regularly arranged in a two-dimensional matrix at a constant pitch. Here, an example of the well plate 11 in which 12×8 (=96) wells W are provided on the plate is shown, but the number of wells provided in one plate is arbitrary.

The well plate 11 may be one in which the base material and the wells W are integrally formed from a resin material, or the wells W formed as separate members are fitted in through-holes provided in the base material. There may be. Alternatively, the structure may be such that the well W is formed by closing the bottom of a cylindrical through-hole provided in the substrate with a separate sheet-like member.

The well plate 11 can store liquid in an internal space (storage space) surrounded by the side wall surfaces and the bottom surface of each well W in such a posture that the upper surface 11a thereof is horizontal. For example, in the fields of medicine and biochemistry, the well plate 11 can be suitably applied for the purpose of culturing cells or the like in the culture solution injected into the wells W to prepare a sample. FIG. 1 shows a state in which the well plate 11 is held in a horizontal posture, with the XY plane representing the horizontal plane and the +Z direction representing the vertically upward direction.

The lid 12 is slightly larger than the well plate 11 and covers the well plate 11 so as to collectively cover the wells W provided in the well plate 11 from above. The lid 12 is provided for the purpose of preventing foreign matter from entering the well W from the outside, temperature change due to evaporation of the culture medium, contact with the outside air, and the like. At least the upper surface of the lid is made of a transparent material such as transparent resin so as not to interfere with optical observation of the sample carried in the well W. The structure of the lid body 12 will be described later in detail.

FIG. 2 is a diagram showing the overall structure of another example of the culture vessel according to the present invention. This culture container 20 includes a dish 21 as a container body and a lid 22 . The dish 21 is, for example, a flat-bottom shallow dish-shaped container made of transparent resin, and is capable of storing liquid in an internal space (storage space) surrounded by side walls and a bottom. The lid 22 is made of a transparent resin and is slightly larger than the dish 21, and has substantially the same structure and function as the lid 12 described above.

FIG. 3 is a cross-sectional view showing the structure of the lid. More specifically, FIG. 3(a) is a diagram showing a cross-sectional structure of the lid body 12 applied to the well plate 11, and FIG. 3(b) is a diagram showing a modification thereof. 3(c) is a cross-sectional view showing the cross-sectional structure of the lid body 22 applied to the dish 21. As shown in FIG. As shown in FIG. 3A, the lid body 12 includes an upper surface portion 120 formed slightly larger than the external size of the well plate 11, and a peripheral portion of the upper surface portion 120 extending downward to the outer peripheral portion of the well plate 11. As shown in FIG. It has an engaging side wall portion 125 .

One of the major features of the lid 12 is that the upper surface portion 120 has a multi-layer structure. Specifically, the upper surface portion 120 has two flat plates 121 and 122 opposed to each other with a predetermined gap therebetween, and a gap space 123 formed between them forms the peripheral edges of the two flat plates 121 and 122 . It has a structure that is airtightly sealed against the external space by a connecting portion 126 that connects them together. The gap space 123 is filled with an inert gas such as dry air or nitrogen gas, and preferably at a pressure lower than atmospheric pressure.

In the following description, in order to distinguish between the two flat plates 121 and 122 forming the upper surface portion 120 of the lid 12, the lid 12 is placed in the inner space of the well plate 11 when placed on the well plate 11. The facing surface, that is, the flat plate 121 on the lower surface side is called a "lower plate". On the other hand, the flat plate 122 on the opposite side, that is, the side facing the external space when placed on the well plate 11 is called the "upper plate".

The reason why the cover 12 has a multi-layer structure is to enhance heat insulation. That is, the culture of cells and the like performed using a culture vessel such as this well plate 11 is performed in an environment of high temperature and high humidity compared to the environment of normal temperature and humidity. For example, culture is performed by placing the well plate 11 in a device called an incubator that realizes such a high-temperature, high-humidity culture environment. In a general culture environment, the temperature is, for example, 37° C. and the humidity is, for example, about 90%.

When the well plate 11 placed in a high-temperature/high-humidity environment as described above is temporarily taken out for the purpose of observing or imaging a sample, for example, in order to prevent foreign matter from entering from the outside and a sudden drop in temperature, , and a lid 12 is attached so as to cover the upper surface thereof. At this time, if water droplets due to dew condensation adhere to the inner surface of the lid 12, it interferes with optical observation. Therefore, it is required to suppress adhesion of water droplets to the lid 12 at least for a certain period of time during which the sample is subjected to observation. In this embodiment, the cover 12 has a multi-layered structure to improve heat insulation, thereby suppressing the occurrence of dew condensation.

As a method for observing or imaging the sample in the culture container with the lid attached, there are a method of observing from above the container and a method of observing from below the container. Here, in the case of observing the sample from above the container, water droplets adhering to the lid can be a direct obstacle to observation. On the other hand, in the case of observation from below, condensation on the lid does not interfere between the observation point and the sample, but it indirectly interferes with observation in that it scatters the illumination light incident on the sample from above. It will be. Therefore, in any case, it is required to prevent dew condensation on the lid in order to perform good observation and imaging.

Making the gap space 123 a reduced-pressure environment also contributes to the improvement of heat insulation. However, if the flat plates 121 and 122 forming the upper surface portion 120 are bent and the gap between them is no longer maintained, the heat insulating effect is reduced. In order to prevent this, a spacer 124 may be provided between the lower plate 121 and the upper plate 122 to maintain a gap, as shown in FIG. 3B, for example. The spacer 124 may be a member separate from the lower plate 121 and the upper plate 122 , or may be a protrusion integrally formed with the lower plate 121 or the upper plate 122 .

The spacer 124 has the function of maintaining the gap between the lower plate 121 and the upper plate 122 even when the gap space is not decompressed. For example, it has the effect of preventing the gap from decreasing due to an external force and the heat insulating performance from deteriorating. In order to suppress heat dissipation via the spacers 124, the number of spacers 124 and the cross-sectional area of each spacer 124 are preferably small, and the spacers 124 are preferably made of a material with low thermal conductivity.

As shown in FIG. 3B, the spacers 124 are desirably provided so as to avoid positions corresponding to the openings of the wells W provided in the well plate 11 . This prevents the spacer 124 from shielding the sample when the sample in the well W is observed.

The lid 22 applied to the sample container 20 can also have the same configuration. That is, as shown in FIG. 3(c), the lid 22 also has two flat plates 221 and 222 facing each other across a predetermined gap, and a gap space 223 formed between them is defined with respect to the external space. The structure can be hermetically sealed. Since the well plate 11 has a substantially rectangular outer shape, the lid body 12 also has a substantially rectangular planar shape, whereas the lid body 22 corresponding to the substantially circular dish 21 has a substantially circular planar shape. .

FIG. 4 is a cross-sectional view showing a configuration example of the lid. The basic structure of the lid 12 covering the upper surface of the well plate 11 is, like the lid 12a shown in FIG. and a connecting portion 126 that keeps the gap constant and seals the gap space 123 . As indicated by the dotted line, by further providing a side wall portion 125 extending downward from the peripheral edge portion of the first flat plate portion 121 , it is possible to define the horizontal position with respect to the well plate 11 .

The lower plate 121, the upper plate 122, and the connecting portion 126 may be integrally formed of the same material, but as a more realistic structure, the lower plate 121 and the upper plate 122 are formed as separate members, and these are assembled together. can be considered to be physically connected. For example, as shown in FIG. 4(b), an upper plate 122 whose periphery extends downward to form a connecting portion 126 may be bonded to the upper portion of a lower plate 121 integrally formed with a side wall portion 125, for example, by adhesion. Thus, the lid body 12b having a multilayer structure can be formed. As the lower plate 121 in this case, it is possible to apply a conventional lid of a single layer structure. Also, the connecting portion 126 may be provided on the upper surface of the lower plate 121 .

Alternatively, for example, as shown in FIG. 4(c), a multilayer structure can be obtained by attaching a lower plate 121 via an appropriate sealing member 127 inside an upper plate 122 that can function as a single-layer lid. lid 12c can be formed. The sealing member 127 has a function of sealing the gap space 123 and a function of a spacer defining the gap.

In addition, this type of culture vessel and the lid used therein may be provided with upwardly protruding ribs on the upper surface of the lid in order to suppress lateral displacement when stacked in multiple stages. . When this rib is provided so as to annularly surround the peripheral edge of the upper surface of the lid, it is possible for this rib to function as a connecting portion that defines the gap. That is, like the lid body 12d shown in FIG. 4D, by attaching the upper plate 122 so as to abut on the annular rib 128 provided on the upper peripheral edge of the lower plate 121, the rib 128 is used as the connecting portion. It becomes possible to seal the gap space 123 while functioning. In this case, if similar ribs 129 are provided on the upper plate 122 like the cover 12e shown in FIG.

As described above, various methods are conceivable for constructing the cover body 12 having a multilayer structure, and any of them may be adopted. Also, the lid 22 corresponding to the dish 21 can have the same structure.

Next, a more detailed structure of the lids 12 and 22, more specifically, a dimensional relationship of each part will be described. Although the lid 12 corresponding to the well plate 11 and the lid 22 corresponding to the dish 21 have different outer shapes in plan view, their structures can be basically the same. Therefore, although the lid 12 corresponding to the well plate 11 will be described in detail below as an example, the lid 22 corresponding to the dish 21 can also be considered in the same way.

BACKGROUND ART Conventionally, a so-called single layer structure in which the upper surface portion is composed of a single flat plate has been widely used as a cover for covering a culture vessel body such as a well plate or dish. When the culture container is covered with such a cover, the culture container may be removed from the culture environment due to the temperature difference between the high-temperature, high-humidity environment of the culture environment and the lower-temperature, normal-temperature environment. Condensation will occur soon after. Water droplets adhering to the upper surface of the lid can be wiped off, but in order to wipe off water droplets adhering to the lower surface, the lid must be removed. For this reason, it is necessary to devise ways to prevent dew condensation from forming on the lower surface of the lid.

By forming the cover into a multi-layered structure as described above to enhance heat insulation, it is possible to delay the occurrence of dew condensation. However, according to the findings of the inventors of the present application, it is not sufficient to simply have a multilayer structure, but the dimensions of each part, specifically, the thickness of the two flat plates 121 and 122 and the size of the gap between them, , depending on the setting, there is a large difference in the dew condensation prevention effect. In the following, the experiments conducted by the inventors of the present application that led to such findings and the preferred dimensions of each part specified based on the experiments will be described.

In the experiment, an appropriate amount of water was injected, and the culture vessel main body (well plate or dish) was warmed by standing for a predetermined time under the same temperature environment as the culture temperature (37°C) in the incubator, and then taken out to the room temperature environment. After that, the cover was attached and the progress was observed to determine the presence or absence of dew condensation on the lower surface of the cover. If the culture container is heated with the lid attached, water droplets may already adhere during the heating process. Therefore, at the timing when the heated container with the lid attached is moved to the room temperature environment, the lid is replaced with a dry lid.

First, an experiment was conducted using an existing single-layer lid. Immediately after placing the normal-temperature lid on the heated culture vessel main body, dew condensation occurred on the lower surface of the lid. If the lid was previously warmed to the same temperature as the culture environment, the time until dew condensation occurred was lengthened, and the dew condensation could be suppressed, for example, by several minutes.

Separately from this, by attaching a resin flat plate to the upper surface of the lid of the multilayer structure shown in FIG. A similar experiment was conducted by preparing a lid having a structure in which the was sealed. Even with such a multi-layered cover, dew condensation occurred immediately after it was placed on the main body of the culture vessel at room temperature. A possible reason for this is that the lower surface of the lid is at a lower temperature than the container body and the liquid inside.

On the other hand, when a multi-layer lid that had been preheated to the culture temperature was attached, the time until dew condensation occurred could be significantly increased. For example, it is possible to suppress the occurrence of dew condensation for about 30 minutes. Considering that the work time is about several minutes for the purpose of observing and imaging the sample in the container, a sufficient dew condensation suppression effect is obtained. It can be said.

From the viewpoint of preventing dew condensation, it is desirable that the two flat plates 121 and 122 be as thick as possible and that the gap between them be as large as possible. However, considering the convenience of accommodating the culture vessel with the lid attached in the incubator or transferring it to various observation devices, the thickness of the lid should not exceed a certain level. desirable. For example, in an observation/imaging apparatus in which one of the illumination means and the light receiving means is arranged above the container and the other is arranged below the container, there is a limit to the height of the culture container that can be handled.

Therefore, the inventors of the present application have studied a lid design rule that can suppress the occurrence of dew condensation for a time necessary and sufficient for observation of a sample under limited dimensional constraints. Specifically, temperature simulations were performed with various combinations of the thickness of the flat plate portion constituting the lid and the size of the gap. We searched for conditions to prevent dew condensation from occurring.

FIG. 5 is a diagram for explaining simulation conditions. As shown in FIG. 5(a), the lid C is modeled as a structure in which two flat plates P1 and P2 having a sufficiently large planar size are arranged facing each other in parallel with a gap G therebetween. was used as a partition wall between the inner space and the outer space of the container, and the change in temperature of each part with time was investigated by simulation.

Here, the thickness of the flat plate (that is, the lower plate) P1 on the side facing the inner space of the container is t1, the thickness of the flat plate (that is, the upper plate) P2 on the side that faces the outer space is t2, and the size of the gap between them is Let g. Further, the surface temperature of the main surface of the lower plate P1 facing the inner space of the container is T1, and the surface temperature of the main surface facing the gap G is T2. Further, let T3 be the surface temperature of the main surface of the upper plate P2 facing the gap G, and T4 be the surface temperature of the side facing the external space.

The temperature Ta1 inside the container was set at 37° C. following the culture environment in the incubator, and the humidity inside the container was set at 90%. Therefore, the initial values of the surface temperatures T1 to T4 are also 37°C. From this temperature and humidity environment, the initial value of the amount of water vapor in the container is estimated to be 39.51 g/cm ³ and the dew point temperature Td is 34.96°C. Also, the temperature Ta2 of the external space was set to 25° C. as a representative numerical value for a room temperature environment.

When temperature simulation is performed under these conditions, as shown in FIG. It will decrease over time by being placed in a normal temperature environment. Compared to the surface temperatures T1 and T2 of the lower P1 inside the container, the surface temperatures T3 and T4 of the upper plate P2 in contact with the external space are significantly lower. Since the temperature differences (T1-T2, T3-T4) between both main surfaces of the flat plates P1 and P2 are small, the temperatures of both main surfaces are represented by one curve in FIG. ing.

When the surface temperature T1 of the lower plate P1 facing the inner space of the container drops to the dew point temperature Td, condensation occurs on the surface. The length D1 of the period until this happens represents the time until dew condensation occurs on the lid.

The following conditions were set as design rules. In view of the current experimental environment using the culture vessel, the thickness of the upper surface of the lid, that is, the sum of the thickness t1 of the lower plate P1, the thickness t2 of the upper plate P2, and the size g of the gap should not exceed 6 mm. do. Moreover, the minimum value of these dimensions was set to 0.5 mm in order to secure the heat insulation performance in each layer. Moreover, the length D1 of the period during which dew condensation does not occur was set to 10 minutes or longer.

When the dimensions of each part satisfying these conditions are obtained, there is a preferable range for the combination of the thicknesses t1 and t2 of the two flat plates, as shown hatched in FIG. 5(c). As the size g of the gap increases, it becomes narrower and shifts toward the lower left in the figure. The preferred range and the area above the dashed line represent exceeding the constraint on the total thickness of the lid. Also, dew condensation occurs under conditions corresponding to the preferable range and the lower left area of the dashed line.

In a more specific temperature simulation, the thermal conductivity of each layer forming the lid affects the results. Here, polystyrene resin and polypropylene resin, which have high light transmittance and can be suitably used for this type of culture vessel, were examined as materials for the upper and lower plates. It was also assumed that the gap was filled with dry air at atmospheric pressure. Some of the calculation results regarding the "preferred range" of the dimensions of each part obtained in this way are shown below.

6 to 9 are diagrams showing the preferable size range of the lid determined from the simulation results. FIG. 6 is a graph showing the results when both the lower plate P1 and the upper plate P2 are made of polystyrene resin, with the thickness t1 of the lower plate P1, the thickness t2 of the upper plate P2, and the size of the gap g as coordinate axes. It is represented by In order to make the drawing easier to see, the coordinate axis representing the size g of the gap is stretched more than the other coordinate axes.

Of these areas, the darkened area is an area that satisfies the conditions that dew condensation does not occur and the thickness of the lid does not exceed the upper limit. It can be said that when a lid made with the relevant dimensional relationships within this area is used, condensation will not occur for at least 10 minutes after being removed from the culture environment. In other words, a lid that does not satisfy these conditions cannot obtain a sufficient anti-condensation effect even if it has a multi-layer structure, or the size exceeds the practically allowable size.

As described above, when viewed on a plane where the gap size g is fixed, the preferable range of the thicknesses (t1, t2) of the two flat plates is from the upper left to the lower right on the plane. As the size g of the gap increases, the range narrows and shifts to the lower left on the plane.

FIG. 7 shows the results when polypropylene resin is used for both the lower plate P1 and the upper plate P2, according to the same rule as in FIG. A preferable range on one plane where the size g of the gap is fixed is wider than that of the polystyrene resin shown in FIG. . It is common to the case of polystyrene resin that the preferable range shifts as the size g of the gap increases.

If the two flat plates forming the lid are formed as different members, it is possible to use a combination of these resin materials. As one of such cases, FIG. 8 shows the results when polypropylene resin is used for the lower plate P1 and polystyrene resin is used for the upper plate P2, and FIG. Similar to the above, the results when polypropylene resin is used for the upper plate P2 are shown.

Although there is some variation in the preferred range of dimensions for each part, the overall trend is common to all materials. By adopting the dimensional relationships based on these calculation results in accordance with the materials used, it is possible to avoid the occurrence of dew condensation on the lid for the time necessary and sufficient to observe the sample. It can be said that this result indicates that it is necessary to appropriately select and apply design rules according to materials, or that it is necessary to appropriately select materials according to dimensional specifications to be satisfied.

The design rules obtained by organizing a series of simulation results are as follows. First, general conditions that do not depend on the materials used are as follows.
- t1, t2 and g are all 0.5 or more;
・ t1 + t2 + g ≤ 6,
must be satisfied. Here, the units of the thickness t1 of the lower plate, the thickness t2 of the upper plate, and the size g of the gap are all in millimeters (mm).

When polystyrene resin is used as the material for both the lower plate and the upper plate, in addition to the above general conditions, the following conditions (1A) to (1C):
(1A) t1+t2≧4.5
(1B) t1+t2≧4.0 and g≧1.5
(1C) t1+t2≧3.5, t1≧1.0, t2≧1.0, and g≧2.5
It is necessary that at least one of

Further, when polypropylene resin is used for both the lower plate and the upper plate, in addition to the above general conditions, the following conditions (2A) to (2G):
(2A) t1+t2≧4.0
(2B) t1+t2≧3.5 and g≧1.0
(2C) t1+t2≧3.5 and t1≧1.5
(2D) t1+t2≧3.5 and t2≧3.0
(2E) t1+t2≧3.0 and g≧2.0
(2F) t1+t2≧3.0, t1≧1.0, t2≧1.0, and g≧1.5
(2G) t1+t2≧2.5 and g≧3.5
It is necessary that at least one of

Further, when a polypropylene resin is used as the lower plate and a polystyrene resin is used as the upper plate, in addition to the above general conditions, the following conditions (3A) to (3J):
(3A) t1+t2≧4.5
(3B) t1+t2≧4.0 and t1≧1.5
(3C) t1+t2≧3.5, t1≧2.0, and g≧1.0
(3D) t1+t2≧3.5 and g≧2.5
(3E) t1+t2≧3.0, t1≧1.5, and g≧2.5
(3F) t1+t2≧3.0, t1≧1.0, and g≧3.0
(3G) t1≧1.5, t2≧2.0, and g≧1.5
(3H) t1≧1.0, t2≧3.0, and g≧1.0
(3I) t1≧1.0, t2≧2.5, and g≧2.0
(3J) t2≧3.5 and g≧1.5
It is necessary that at least one of

Further, when a polystyrene resin is used as the lower plate and a polypropylene resin is used as the upper plate, in addition to the above general conditions, the following conditions (4A) to (4J):
(4A) t1+t2≧4.5
(4B) t1+t2≧4.0 and t2≧1.5
(4C) t1+t2≧4.0 and g≧1.5
(4D) t1+t2≧3.5 and t2≧2.0,
(4E) t1+t2≧3.5, t2≧1.5, and g≧1.5
(4F) t1+t2≧3.5, t2≧1.0, and g≧2.0
(4G) t1+t2≧3.5 and g≧2.5
(4H) t1+t2≧3.0, t2≧1.5, and g≧2.5
(4I) t1+t2≧3.0, t2≧1.0, and g≧3.0
(4J) t1≧3.0 and t2≧1.0
It is necessary that at least one of

In addition to the above general conditions, the following conditions are applicable as design rules that can be established regardless of whether polystyrene resin or polypropylene resin is used for the upper and lower plates:
・t1+t2≧4.5
must be established.

According to these conditions, it is possible to manufacture a lid that has an upper surface portion thickness of 6 mm or less and that can prevent dew condensation for at least 10 minutes after removal from the culture environment. When the culture vessel is removed from the culture environment, by attaching the cover thus manufactured, it is possible to observe and image the sample in the culture vessel without being affected by condensation. becomes.

In particular, it is effective to increase the values of t1, t2, and g as much as possible within the range corresponding to the above conditions, in order to further improve the heat insulating properties. As can be seen from the graph in FIG. 5(b), it is the gap space that exhibits a particularly high heat insulation effect, so it is desirable to make the size g of the gap as large as possible. For example, when the temperature of the external space is lower than 25° C., the period during which dew condensation does not occur may be shorter than the above. Enlarging the dimensions of each part as much as possible is also effective in avoiding unexpected dew condensation under such conditions.

For example, in the lid body 12 used in combination with the well plate 11, the above conditions are applied with the thickness of the lower plate 121 as t1, the thickness of the upper plate 122 as t2, and the size of the gap between them as g. be able to. Further, for example, in the lid body 22 used in combination with the dish 21, the above conditions are applied with the thickness of the lower plate 221 as t1, the thickness of the upper plate 222 as t2, and the size of the gap between them as g. can do.

Note that even with a lid that meets the above conditions, dew condensation may occur at an earlier stage depending on how it is used. That is, when a lid having a temperature lower than that of the culture vessel main body and its contents is attached, water vapor cooled near the surface of the lid causes dew condensation. To prevent this, the temperature of the lid should be approximately the same as the culture temperature when it is removed from the culture environment. For example, by storing the lid together with the main body of the culture vessel in the incubator, or by using a separate heating device, the lid is preheated to about the culture temperature, and is warmed when the main body of the vessel is taken out. By installing the lid body, the dew condensation suppression effect described above can be stably obtained.

The culture container body may be stored in the incubator with the lid attached, but since water droplets may adhere during storage, a new dry lid should be attached when the culture vessel is taken out. is more preferred. In this case, for example, the lid may be replaced in the culture environment before taking out, or the lid may be replaced immediately after taking out. Further, for example, a single-layer lid may be attached to the container body in the incubator, and replaced with a preheated multi-layer lid when it is taken out.

As described above, in the above embodiment, the well plate 11 and the dish 21 correspond to the "culture vessel main body" of the present invention, and the lids 12 and 22 correspond to the "culture vessel lid" of the present invention. ing. The set of well plate 11 and lid 12 and the set of dish 21 and lid 22 each correspond to the "culture vessel" of the present invention.

The lower plate 121 of the lid 12 and the lower plate 221 of the lid 22 respectively correspond to the "first flat plate portion" of the present invention. correspond to the "second flat plate portion" of the present invention, and the structure connecting them corresponds to the "connecting portion" of the present invention.

The present invention is not limited to the above-described embodiments, and various modifications other than those described above can be made without departing from the spirit of the present invention. For example, the lids 12 and 22 of the above-described embodiments have side wall portions extending downward from the periphery of the upper surface portion, and are attached to the culture vessel main body so as to cover the upper surface and side surfaces thereof. However, the design according to the present invention can be applied even to a lid body which is simply placed on the upper surface of the container body as conceptually shown in FIG. A rule-based multilayer structure is effective in preventing dew condensation. Also in this case, it is preferable that either the container main body or the lid is provided with a protrusion or a groove or the like for restricting mutual positional deviation.

In addition, in the description of the above embodiments, a well plate and a dish are used as examples of the culture vessel main body. However, the culture vessel body used in combination with the lid according to the present invention is not limited to these, and the present invention is applied to lids that are combined with various shaped vessel bodies used as culture vessels. Is possible.

In addition, although the simulation conditions and lid constituent materials used in the above embodiments are in line with the actual conditions of research sites where such culture vessels are used, these conditions may be partially changed as necessary. It is also possible to change and perform simulations to formulate new design rules.

As described above by exemplifying specific embodiments, in the lid for a culture vessel according to the present invention, one of the first flat plate portion and the second flat plate portion and the connecting portion are integrally molded. It can be anything. According to such a configuration, it is possible to configure the lid for a culture vessel essentially with two parts, and it is possible to provide a lid for a culture vessel with a simple manufacturing process and at a low cost. .

Further, for example, the gap space may be filled with dry gas. According to such a configuration, it is possible to prevent condensation from occurring on the surfaces of the first flat plate portion and the second flat plate portion facing the inside of the gap space.

Further, for example, the air pressure in the gap space may be configured to be lower than the atmospheric pressure. According to such a configuration, heat conduction through the gas in the gap space is suppressed, and heat insulation can be further improved.

Further, for example, a spacer may be provided in the gap space between the two flat plate portions to define the minimum value of the gap. For example, a protrusion that is provided on at least one of the two flat plate portions and protrudes toward the other flat plate portion may function as a spacer that defines the minimum value of the gap by contacting the other flat plate portion. can be done. According to such a configuration, it is possible to prevent the gap from being reduced due to deformation of one of the two flat plate portions due to flexure or the like, thereby reducing the heat insulating property. In particular, when one of the two flat plate portions is thin, or when the gap space is a pressure-reduced space, it is effective to take measures to prevent such reduction of the gap.

Here, when the culture vessel main body combined with the culture vessel lid according to the present invention is a well plate having a plurality of wells each capable of holding a liquid, the culture vessel lid covers the well plate. It is preferable that the protrusion is provided at a position that does not overlap with the well in plan view when placed. With such a configuration, it is possible to prevent the protrusion from becoming an obstacle when observing the sample in the well.

INDUSTRIAL APPLICABILITY The present invention can be applied to general sample containers that hold culture objects such as cells together with liquid. In particular, it is suitable for the purpose of subjecting samples prepared by culturing to imaging. Such a sample container can be suitably applied in fields such as medicine, biochemistry and drug discovery.

10, 20 culture vessel 11 well plate (main body of culture vessel)
12, 22 lid body (lid body for culture vessel)
21 dish (culture vessel body)
121, 221 lower plate (first flat plate portion)
122, 222 upper plate (second flat plate portion)
124 Spacer 126 Connection g Gap size t1 Lower plate thickness t2 Upper plate thickness W Well (storage space)

Claims (11)

A culture vessel lid covering the upper part of a culture vessel body having an open top and a storage space capable of storing liquid,
two flat plate portions each made of a transparent resin and having a planar size corresponding to the planar size of the culture vessel main body;
By connecting the peripheral edges of the two flat plate portions to each other, the two flat plate portions are opposed to each other with a predetermined gap therebetween, and the gap space between the two flat plate portions is airtight with respect to the external space. and a connection that seals to
Of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, and the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm], the thickness of the second flat plate portion is t2 [mm], and the size of the gap is g [mm] (where t1, t2, and g are all 0.5 or more, and t1+t2+g≦6),
Both the first flat plate portion and the second flat plate portion are made of polystyrene resin,
Conditions below:
(1A) t1+t2≧4.5
(1B) t1+t2≧4.0 and g≧1.5
(1C) t1+t2≧3.5, t1≧1.0, t2≧1.0, and g≧2.5
At least one of the culture container lids is established. A culture vessel lid covering the upper part of a culture vessel body having an open top and a storage space capable of storing liquid,
two flat plate portions each made of a transparent resin and having a planar size corresponding to the planar size of the culture vessel main body;
By connecting the peripheral edges of the two flat plate portions to each other, the two flat plate portions are opposed to each other with a predetermined gap therebetween, and the gap space between the two flat plate portions is airtight with respect to the external space. and a connection that seals to
Of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, and the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm], the thickness of the second flat plate portion is t2 [mm], and the size of the gap is g [mm] (where t1, t2, and g are all 0.5 or more, and t1+t2+g≦6),
Both the first flat plate portion and the second flat plate portion are made of polypropylene resin,
Conditions below:
(2A) t1+t2≧4.0
(2B) t1+t2≧3.5 and g≧1.0
(2C) t1+t2≧3.5 and t1≧1.5
(2D) t1+t2≧3.5 and t2≧3.0
(2E) t1+t2≧3.0 and g≧2.0
(2F) t1+t2≧3.0, t1≧1.0, t2≧1.0, and g≧1.5
(2G) t1+t2≧2.5 and g≧3.5
At least one of the culture container lids is established. A culture vessel lid covering the upper part of a culture vessel body having an open top and a storage space capable of storing liquid,
two flat plate portions each made of a transparent resin and having a planar size corresponding to the planar size of the culture vessel main body;
By connecting the peripheral edges of the two flat plate portions to each other, the two flat plate portions are opposed to each other with a predetermined gap therebetween, and the gap space between the two flat plate portions is airtight with respect to the external space. and a connection that seals to
Of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, and the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm], the thickness of the second flat plate portion is t2 [mm], and the size of the gap is g [mm] (where t1, t2, and g are all 0.5 or more, and t1+t2+g≦6),
The first flat plate portion is made of polypropylene resin, the second flat plate portion is made of polystyrene resin,
Conditions below:
(3A) t1+t2≧4.5
(3B) t1+t2≧4.0 and t1≧1.5
(3C) t1+t2≧3.5, t1≧2.0, and g≧1.0
(3D) t1+t2≧3.5 and g≧2.5
(3E) t1+t2≧3.0, t1≧1.5, and g≧2.5
(3F) t1+t2≧3.0, t1≧1.0, and g≧3.0
(3G) t1≧1.5, t2≧2.0, and g≧1.5
(3H) t1≧1.0, t2≧3.0, and g≧1.0
(3I) t1≧1.0, t2≧2.5, and g≧2.0
(3J) t2≧3.5 and g≧1.5
At least one of the culture container lids is established. A culture vessel lid covering the upper part of a culture vessel body having an open top and a storage space capable of storing liquid,
two flat plate portions each made of a transparent resin and having a planar size corresponding to the planar size of the culture vessel main body;
By connecting the peripheral edges of the two flat plate portions to each other, the two flat plate portions are opposed to each other with a predetermined gap therebetween, and the gap space between the two flat plate portions is airtight with respect to the external space. and a connection that seals to
Of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, and the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm], the thickness of the second flat plate portion is t2 [mm], and the size of the gap is g [mm] (where t1, t2, and g are all 0.5 or more, and t1+t2+g≦6),
The first flat plate portion is made of polystyrene resin, the second flat plate portion is made of polypropylene resin,
Conditions below:
(4A) t1+t2≧4.5
(4B) t1+t2≧4.0 and t2≧1.5
(4C) t1+t2≧4.0 and g≧1.5
(4D) t1+t2≧3.5 and t2≧2.0,
(4E) t1+t2≧3.5, t2≧1.5, and g≧1.5
(4F) t1+t2≧3.5, t2≧1.0, and g≧2.0
(4G) t1+t2≧3.5 and g≧2.5
(4H) t1+t2≧3.0, t2≧1.5, and g≧2.5
(4I) t1+t2≧3.0, t2≧1.0, and g≧3.0
(4J) t1≧3.0 and t2≧1.0
At least one of the culture container lids is established. A culture vessel lid covering the upper part of a culture vessel body having an open top and a storage space capable of storing liquid,
two flat plate portions each made of a transparent resin and having a planar size corresponding to the planar size of the culture vessel main body;
By connecting the peripheral edges of the two flat plate portions to each other, the two flat plate portions are opposed to each other with a predetermined gap therebetween, and the gap space between the two flat plate portions is airtight with respect to the external space. and a connection that seals to
Of the two flat plate portions, one of the flat plate portions facing the storage space is a first flat plate portion, and the other flat plate portion is a second flat plate portion, and the thickness of the first flat plate portion is t1 [mm], the thickness of the second flat plate portion is t2 [mm], and the size of the gap is g [mm], the following conditions:
(5A) t1, t2 and g are all 0.5 or more;
(5B) t1+t2+g≦6,
(5C) t1+t2≧4.5
A cover for a culture vessel that satisfies all of the above. 6. The lid for culture vessel according to claim 1, 2 or 5, wherein one of said first flat plate portion and said second flat plate portion and said connecting portion are integrally molded. 7. The culture vessel lid according to any one of claims 1 to 6, wherein said gap space is filled with dry gas. 8. The culture vessel lid according to any one of claims 1 to 7, wherein the air pressure in said gap space is lower than atmospheric pressure. 9. The culture vessel lid according to any one of claims 1 to 8, wherein a spacer is provided between the two flat plate portions in the gap space to define the minimum value of the gap. 10. The culture vessel cover according to any one of claims 1 to 9, which covers the upper part of the culture vessel body, which is a well plate having a plurality of wells each capable of holding a liquid,
At least one of the two flat plate portions is provided with a protrusion that protrudes toward the other flat plate portion and defines the minimum value of the gap,
A lid for a culture vessel, wherein the protrusion is provided at a position that does not overlap the well in plan view when the lid for the culture vessel is placed covering the well plate. a culture vessel main body having an upper opening and a storage space capable of storing liquid;
A culture vessel comprising the lid for a culture vessel according to any one of claims 1 to 10.

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