JP5084187B2 - Incubator and humidifier dish for incubator - Google Patents

Description

  The present invention relates to an incubator having a gas chamber, a temperature, and a humidity environment suitable for culturing a culture chamber, and a humidifying dish used in the incubator.

  With the recent development of fields related to biotechnology and regenerative medicine, the work of culturing cells using an incubator is increasing. In order to promote cell culture, it is necessary to prepare a culture space suitable for each cell. Conventionally, incubators that perform temperature control, humidity control, and atmosphere control of the culture space have been developed.

In particular, when culturing cells that require a strict concentration condition of CO ₂ (carbon dioxide) gas concentration as a culture condition, in addition to the temperature control and humidity control, the CO ₂ gas concentration in the culture space is set. A controllable CO ₂ incubator is used.

In addition, when culturing cells that require a strict concentration condition of O ₂ (oxygen) gas concentration as a culture condition, in addition to controlling temperature and humidity, the O ₂ gas concentration in the culture space is controlled. A possible incubator is used. Generally, when adjusting the concentration of O ₂ gas, when the set O ₂ gas concentration is less than 20%, O ₂ gas is supplied from the O ₂ gas cylinder to the culture space, and the set O ₂ gas concentration is 20 In the case of% or less, N ₂ gas is supplied from the N ₂ gas cylinder to the culture space.

In the culture space, in order to promote the culture of cells and the like, control is performed to maintain the humidity at a high level, for example, 90% or more (see, for example, Patent Document 1 and Patent Document 2). As humidification means for the culture space described in each of these patent documents, a humidification tray in which water is stored is installed on the bottom surface of the culture space, and the culture space is humidified by moisture naturally evaporated from the humidification tray. It was.
JP-A-10-234351 JP 2002-112762 A

  However, in the method of humidifying the culture space by natural evaporation from the humidifying tray, it depends on the amount of evaporation from the humidifying tray due to the relationship with the temperature of the culture space. Therefore, when the humidity in the culture space becomes 95% or more, a slight temperature difference (for example, 0.5 ° C.) between the inner wall surface and shelf surface constituting the culture space and the sample container in which the culture is accommodated. When this occurs, there is a problem that condensation occurs on the lower surface.

  In particular, in an incubator equipped with a microscope function, a temperature difference occurs between the table surface on which the culture is observed and the culture space, and the inner surface and outer surface of the sample container in which the lens constituting the microscope and the observation target are accommodated. When dew condensation occurs in the medium, there is a problem that the culture cannot be observed with a microscope.

Therefore, by letting out air in the culture space and introducing a new gas (CO ₂ gas, O ₂ gas or N ₂ gas, outside air), humidity (high humidity and humidity that does not easily cause condensation in the culture space ( For example, it is conceivable to maintain at 90%.

However, O ₂ gas and N ₂ gas are relatively expensive. Therefore, it is preferable to adjust the O ₂ gas concentration according to the minimum necessary gas introduction amount, and development of a mechanism for adjusting the humidity in the culture space to a predetermined high humidity by a method other than gas replacement is desired. Yes.

  Therefore, the present invention has been made to solve the conventional technical problems, makes it possible to control the amount of evaporation from a humidifying dish in which humidifying water is stored, and to prevent the occurrence of condensation in the culture space. It is an object of the present invention to provide an incubator that can be suppressed and a humidifying dish used in the incubator.

A humidifying dish for an incubator according to the present invention is a humidifying dish that is configured by a member having good heat conductivity and is provided in a culture chamber of an incubator to store water for humidification . A lid provided on the upper surface opening, the lid comprising a window for exposing humidification water in the culture chamber, and a movable blocking member for adjusting the area of the water surface exposed in the window, The movable closing member has a first closing portion that is formed at the lower edge of the window except the side positioned in the opening direction of the movable closing member and hangs down toward the bottom surface of the humidifying dish, and the movable closing member slides on the upper surface of the lid. The first and second closing portions have a top surface portion that moves and a second closing portion that hangs down from the end of the top surface portion toward the bottom surface of the humidifying dish, and slides the top surface portion of the movable closing member. It is possible to adjust the area of the water surface exposed to the culture chamber surrounded by And features.

According to a second aspect of the present invention, in the above invention, the lid has a gas introduction portion located outside the window.

An incubator according to a third aspect of the present invention is provided with the humidifying dish for an incubator according to the first or second aspect in a culture chamber, wherein the culture chamber has a gas, temperature and humidity environment suitable for culture of the culture. And

According to the present invention, the humidifying dish for an incubator is a humidifying dish that is configured by a member having good heat conduction and is provided in a culture chamber of an incubator to store water for humidification . A lid provided on the upper surface opening of the main body, and the lid is capable of adjusting the area of the water surface exposed in the culture chamber, so that the amount of evaporation from the water surface exposed in the culture chamber can be controlled by opening and closing the lid. It becomes possible to control by.

  Thereby, when the humidity in the culture chamber rises above a predetermined humidity and an environment in which condensation is likely to occur occurs, by limiting the area of the water surface exposed to the culture chamber of the humidifying dish by the lid, It becomes possible to limit the amount of evaporation from the humidifying tray. Therefore, it is possible to suppress the occurrence of condensation.

Further, since the lid can adjust the area of the water surface exposed in the culture chamber, the area of the water surface exposed in the culture chamber can be arbitrarily adjusted, and the area of the water surface exposed in the culture chamber Can be finely adjusted. This allows the culture chamber to be adjusted to the desired humidity compared to when the water surface exposed to the culture chamber is completely blocked by the lid or when the water in the humidifying dish is completely exposed to the culture chamber. It becomes easy to do.

  As a result, it is possible to easily adjust the humidity, and it is possible to maintain high humidity suitable for culturing the culture while suppressing the occurrence of condensation.

In particular, the lid is positioned in the opening direction of the window for exposing the humidifying water into the culture chamber, a movable blocking member for adjusting the area of the water surface exposed in the window, and the movable blocking member. A first closing portion formed on a lower edge of the window excluding the side and hanging down toward the bottom surface of the humidifying dish, and the movable closing member includes an upper surface portion that slides on the upper surface of the lid, and an upper surface portion of the upper surface portion. It has a second closing part that hangs down from the end toward the bottom of the humidifying dish, and the area of the water surface surrounded by the first and second closing parts can be adjusted by sliding the upper surface part of the movable closing member. Therefore, the area of the water surface exposed in the culture chamber can be finely adjusted with a simple structure.

  This also makes it possible to easily adjust the humidity and maintain high humidity suitable for culturing the culture while suppressing the occurrence of condensation.

According to the invention of claim 2, in addition to the above-mentioned invention, the lid has a gas introduction part located outside the window, so that gas can be introduced into the water for humidification stored in the humidification dish. It becomes. As a result, the gas introduced into the culture chamber can be once humidified with the water for humidification and then diffused into the culture chamber, so that the humidification efficiency can be improved.

  In particular, since the gas introduction part is located outside the window of the lid, it is possible to suppress the inconvenience that water splash caused by the gas introduced into the humidifying water leaks to the outside through the window. It becomes.

And according to invention of Claim 3, in the incubator which makes the culture room the gas suitable for culture | cultivation of culture, temperature, and humidity environment, since the humidification dish for incubators of said each invention is provided in the culture room, By having a lid for limiting the water surface exposed in the culture chamber, the evaporation amount from the water surface exposed in the culture chamber can be adjusted by opening and closing the lid.

  Thereby, when the humidity in the culture chamber rises above a predetermined humidity and an environment in which condensation is likely to occur occurs, by limiting the area of the water surface exposed to the culture chamber of the humidifying dish by the lid, It becomes possible to limit the amount of evaporation from the humidifying tray. Therefore, it is possible to suppress the occurrence of condensation.

  Next, embodiments of the present invention will be described in detail with reference to the drawings. 1 is a partially transparent front view of a culture observation apparatus 1 constituting a culture observation system S to which an incubator of the present invention is applied, FIG. 2 is a side view of the culture observation apparatus 1 of FIG. 1, and FIG. 4 is a longitudinal side view of the culture observation apparatus 1, FIG. 4 is a transverse plan view of the culture observation apparatus 1 in FIG. 1, and FIG. 5 is an electric block diagram of a control device constituting the culture observation system S.

  The culture observation system S of the present embodiment is, for example, an embryonic stem cell that is an undifferentiated stem cell (so-called ES cell) existing in a fertilized egg, or an undifferentiated existing in each organ in the body as a culture. System used for culturing not only cells used for regenerative medicine such as somatic stem cells such as hematopoietic stem cells and neural tube cells, but also cells such as fertilized eggs. It is a system that makes it possible to observe.

  The culture observation system S includes an incubator (culture means) 2 for configuring an environment suitable for culturing a cell as a culture (hereinafter, described by taking a cell as an example of the culture), and a microscopic image of the cell. An image pickup apparatus (image pickup means) 3 for taking a picture, a display (display means) 4 for displaying a microscope image taken by the image pickup apparatus 3, and a control for controlling the incubator 2, the image pickup apparatus 3 and the display 4 And a computer 5 as means. Among these, the culture observation apparatus 1 as shown in FIG. 1 is comprised by the incubator 2 and the imaging device 3, and the computer 5 is comprised from PC etc. which were comprised by the general purpose microcomputer.

  In the culture observation apparatus 1, a main body 12 is constituted by, for example, a heat-insulating box having an opening 10 on one surface (front surface in the present embodiment), and an incubator 2 is provided in an upper portion of the main body 2 (inside the warehouse). A culture chamber 13 is formed. An imaging chamber 6 in which the imaging device 3 is disposed is formed in the lower part of the main body 12 positioned below the culture chamber 13.

  The main body 12 is provided with an inner door 14 for closing the front opening 10 so as to be openable and closable, and further, a heat insulating door 15 is provided outside the inner door 14. The inner door 14 is made of a transparent glass plate or the like that allows the inside to be seen through, and the inside of the culture chamber 13 can be visually confirmed with the heat insulating door 15 opened and the inner door 14 closed. A control panel 16 is provided on the front surface of the heat insulating door 15, and a gasket 17 is attached to the outer peripheral edge of the back surface of the heat insulating door 15 so as to be located outside the end surface of the inner door 14. Thereby, the culture chamber 13 can be sealed from the outside.

  On the inner wall surface of the main body 12 constituting the culture chamber 13, a thermally conductive partition wall 20 is disposed at a predetermined interval. The partition wall 20 is disposed on the entire surface constituting the culture chamber 13, that is, the top surface, the bottom surface, the left and right side surfaces, and the back surface, and between the partition wall 20 and the inner wall surface of the main body 12, An in-compartment heater 19 (only FIG. 5 is shown) is provided. In addition, since the imaging device 3 is disposed below the culture chamber 13, the inside heater 19 is disposed on the bottom surface of the culture chamber 13 except for the portion where the imaging device 3 is provided. It shall be. In addition, a door heater 18 (shown only in FIG. 5) is also disposed on the back surface of the heat insulating door 15 constituting the front surface of the culture chamber 13 with the door 15 closed. Thereby, the culture chamber 13 can be heated from all six surfaces indirectly through the partition wall 20 and the inner door 14.

  Further, each partition wall 20 constituting the left and right side walls of the culture chamber 13 is formed with a plurality of stages of shelf racks (not shown) that project inward, and the shelf racks have thermal conductivity, and A plurality of shelves (shelves) 7 in which a plurality of communication holes are formed are detachably mounted in a plurality of stages (six stages in this embodiment). As will be described in detail later, since the light source 47 is disposed in the rear part of the culture chamber 13, in this embodiment, the rear end of the shelf plate 7 is installed so as to be in front of the front surface of the light source 47. However, the present invention is not limited to this, and extends to a position close to the back surface constituting the culture chamber 13, and the position corresponding to the light source 47 has a shape (cross section U) that avoids the light source 47. It may be a letter-shaped notch.

On the other hand, the main body 12 is provided with a CO ₂ gas supply port 23 and an O ₂ · N ₂ gas supply port 21 so as to communicate with the inside of the culture chamber 13. A gas supply pipe (not shown) is connected. The gas supply pipe connected to the CO ₂ gas supply port 23, via the CO ₂ gas electromagnetic valve 24 CO ₂ gas cylinder (CO ₂ gas supply means) is connected.

Further, the gas supply pipe connected to the O ₂ · N ₂ gas supply port 21, through the O ₂ gas electromagnetic valve 25 through the O ₂ gas cylinder (O ₂ gas supply means) and the N ₂ gas electromagnetic valve 22 An N ₂ gas cylinder (N ₂ gas supply means) is connected. In this embodiment, each O ₂ gas and N ₂ gas is supplied to the culture chamber 13 through the O ₂ · N ₂ gas supply port 21, but the present invention is not limited to this, and similarly O ₂ gas. and N ₂ gas for supplying to the N ₂ gas in the cultivation room 13 further provided one anymore O ₂ · N ₂ gas inlet, thereby, may be as smooth gas supply.

Each gas cylinder is assumed to contain a gas having a predetermined purity or higher. Each electromagnetic valve 22, 24, 25 is controlled to be opened and closed by an incubator-side control device C1 described later in detail. As a result, a gas having a predetermined CO ₂ concentration and O ₂ concentration can be supplied to the culture chamber 13. In the present embodiment, both O ₂ gas and N ₂ gas can be supplied, but only one of them may be supplied.

Further, the main body 12, the inside temperature sensor 26 for detecting the air temperature in the culturing room 13, the CO ₂ gas concentration sensor 27 for detecting the CO ₂ gas concentration in the culturing room 13, the culture chamber 13 O ₂ gas concentration sensor 28 for detecting the O ₂ gas concentration is provided. These sensors 26, 27, and 28 are all connected to the incubator side control device C1, and based on the detection of these sensors, the internal heater 19, the door heater 18, the CO ₂ gas electromagnetic valve 24, and the O ₂ gas electromagnetic valve. The valve 25 and the N ₂ gas electromagnetic valve 22 are controlled, and the temperature and gas concentration of the culture environment set as will be described in detail later are controlled.

  The main body 12 is provided with a sample collection port 29 for taking out the air in the culture chamber 13 as a sample. In addition, a blower (not shown) is provided in the main body 12 for stirring the air in the culture chamber 13 and making the air state uniform. This blower is operated by a blower motor 32, and the blower motor 32 is controlled by an incubator-side control device C1.

A gas introduction port 30 formed in communication with the O ₂ · N ₂ gas supply port 21 is formed on the culture chamber 13 side of the O ₂ · N ₂ gas supply port 21 as described above. The gas introduction port 30 is connected to the air introduction port 58 of the humidifying tray 55 via the gas pipe 54.

  Here, with reference to FIG. 6 thru | or FIG. 10, the structure of the humidification tray 55 is demonstrated. 6 is a vertical side view of the humidifying tray 55, FIG. 7 is a plan view of the humidifying tray 55 of FIG. 6, FIG. 8 is a vertical front view of the humidifying tray 55 of FIG. Fig. 9 shows a longitudinal front view of the lid 60 of Fig. 9.

  The humidifying dish 55 in the present embodiment is placed on the uppermost shelf 7 installed in the culture chamber 13 and includes a main body 56 that opens to the upper surface and a lid 60 that closes the upper surface opening. Is done. The main body 56 and the lid 60 are made of an antibacterial treatment and a member having good heat conduction, for example, a metal material.

  The main body 56 is configured in a container shape having a predetermined volume, and is capable of storing humidifying water, for example, distilled water. In addition, in the state where the lid 60 which will be described later is not attached, all water surfaces of the humidifying water stored in the main body 56 are exposed in the culture chamber 13.

  The lid 60 is formed with a window 61 for exposing the humidifying water stored in the main body 56 to the inside of the culture chamber 13, and the window 61 is a movable closing member made of the same material. 62 is slidably mounted. Further, the lid 60 is provided with an air introduction port (gas introduction part) 58 for connecting a gas pipe 54 whose one end is connected to the gas introduction port 30, located outside the window 61. In the present embodiment, the side on which the air introduction port 58 is provided is disposed on the back side of the culture chamber 13, and the movable closing member 62 closes the window 61, so that the air introduction port 58 is opposite to the direction opposite to the air introduction port 58. The window 61 can be opened by sliding forward.

  Here, for example, the lower edge portion (three surfaces in this embodiment) of the window 61 formed on the lid 60 excluding the side located in the opening direction of the movable closing member 62 is suspended toward the bottom surface of the main body 56. A blocking surface 57 (first blocking portion) is formed. The closing surface 57 in the present embodiment is formed to have a dimension that allows a water passage to be formed between the bottom surface (humidifying tray bottom surface) of the main body 56, the projection surface of the window 61, and The water surface in the main body 56 other than the surface opened by the movable closing member 62 is not exposed to the culture chamber 13.

  The movable closing member 62 is a plate-like member bent in a substantially L-shaped cross section, and has an upper surface 62A (upper surface portion) positioned substantially horizontally with respect to the upper surface of the lid 60, and an end portion of the upper surface 62A. 56 is composed of a blocking surface 62B (second blocking portion) formed by hanging down toward the bottom surface. Thereby, the surface on which the closing surface 57 is not formed is also the projection surface of the window 61 and the surface opened by the movable closing member 62 by the closing surface 62B formed on the movable closing member 62. It can be set as the structure which the water surface in the main body 56 located other than is not exposed in the culture chamber 13. FIG.

  Here, the upper surface 62A is wider than the width dimension of the window 61 formed in the lid 60 (dimension in the direction orthogonal to the sliding direction of the movable closing member 62), specifically, the opening edge of the window 61. The size is such that it can be stably placed on top. At least one side, that is, the side extending in the sliding direction, overlaps with the engagement hole 63 formed in advance on the edge of the window 61 of the lid 60 and can be screwed with a screw (not shown). A plurality of engaging holes 64 are formed at predetermined intervals.

  The closing surface 62B of the movable closing member 62 is formed to be slightly smaller than the width dimension of the window 61 to be inserted (that is, the dimension in the direction orthogonal to the sliding direction of the movable closing member 62). The lower end of the closing surface 62B is formed to be slightly shorter than the lower end of the closing surface 57 formed in the window 61 of the lid 60. Thus, even when the movable closing member 62 is slid back and forth with the movable closing member 62 attached to the window 61, the closing surfaces 62B of the movable closing member 62 come into contact with the closing surfaces 57 located at both ends. It is possible to suppress the generation of unpleasant contact sounds and the occurrence of scratches caused by contact.

  On the other hand, as shown in FIG. 10, the air introduction port 58 formed in the lid 60 is constituted by a tubular member having a substantially crank-shaped cross section, and one end is located above the lid 60 and the other end is in the container 56. To position. Both ends are formed so as to open, and the air introduced from one end can be discharged from the other end. In addition, the air introduction port 58 in this embodiment has a lower end, that is, an end located in the container 56, located at the lower part of the container 56, and a small amount of humidifying water stored in the container 56. Even in such a case, the air introduced from one end of the air introduction port 58 can be discharged into the water.

  Then, a predetermined amount of water for humidification is stored in the container 56 and installed in the culture chamber 13 with the lid 61 attached. When the humidity in the culture chamber 13 does not reach a value suitable for a predetermined culture condition, for example, when the humidity is at least 90% or less, the movable closing member 62 is pulled forward and the window 61 is fully opened. And

  On the other hand, in the culture chamber 13 formed in the main body 12, a table constituting the imaging device 3 together with a lens 35 such as a magnification changing lens, a CCD camera 36, etc., which are arranged in the imaging chamber 6 described later in detail. 37 is disposed. The table 37 is a plate-like member that is made of a heat-conducting material and has a substantially circular plane in this embodiment as shown in FIG. The table 37 is formed with a plurality of holding holes 43 formed so as to communicate with each other in order to install a plurality of sample containers 42 containing samples such as cells. In the present embodiment, six holding holes 43 are formed with a predetermined interval, and these holding holes 43 are all formed with a predetermined dimension from the center of the table.

  The sample container 42 is made of a light-transmitting container having at least a bottom surface and a top surface, for example, transparent glass or resin, and transmits the irradiation light from the light source 47 and observes the contents with the objective lens 44. It is supposed to be possible. Further, the opening of the sample container 42 can be closed with a lid member provided with a breathable filter, and the humidity and gas concentration in the container 42 can be made the same as in the culture chamber 13.

  A rotating shaft 38 made of a heat conductive material is attached to the center of the table 37. One end of the rotary shaft 38 is connected to a rotational movement stage motor 39 (only FIG. 5 is shown) disposed in the imaging chamber 6, and can be rotated in the horizontal direction by driving the stage motor 39. . The rotating shaft 38 can be moved in one direction, in this embodiment, in the front-rear direction by a stage motor 40 for linear movement. Thereby, the cell etc. in the sample container 42 installed in the table 37 can be moved by moving the table 37 by rotational movement and linear movement.

  A stage heater 41 is attached to the rotary shaft 38, and the heat conductive table 37 attached to one end of the rotary shaft 38 can be heated to a predetermined temperature. A stage temperature sensor 46 for detecting the temperature of the table 37 is attached to the table 37.

  The culture chamber 13 in which the table 37 is disposed and the imaging chamber 6 in which the imaging device 3 is disposed are partitioned by a partition wall 45, and the adjusted air in the culture chamber 13 is imaged. A sealed structure is adopted so as not to enter the chamber 6.

  An objective lens 44 is provided below the table 37 so as to protrude from the imaging chamber 6 side through the partition wall 45. The objective lens 44 enables observation of cells in the sample container 42 installed in the holding hole 43 of the table 37, and a position at a predetermined interval from the rotating shaft 38 (table center), that is, holding. It is provided at a position corresponding to the position where the hole 43 is formed. At a position facing the objective lens 44 (above the objective lens 44), a light source 47 constituting the imaging device 3 is provided on the main body 12 located in the culture chamber 13 via an arm 48.

  The light source 47 in the present embodiment uses LED illumination, extends vertically in the culture chamber 13, and is located between the objective lens 44 and the irradiation hole of the light source 47 from above. Illuminate etc. The light source 47 is not limited to LED illumination, and may be a mercury lamp or an optical fiber, for example.

  In the present embodiment, since a plurality of stages of shelves 7 can be installed in the culture chamber 13 above the table 37, the light source 47 is a culture (contained in the container) that is performed from the front opening 10. In the present embodiment, the cell is placed at the rear of the culture space so as not to interfere with the delivery operation of the accommodated cells. Similarly, the objective lens 44 provided to face the irradiation hole of the light source 47 is also disposed at the rear of the culture space.

  The objective lens 44, the light source 47, the table 37, the magnification changing lens 35, the CCD camera 36, and the like constituting the imaging apparatus 3 are disposed in the imaging chamber 6 together with the motors 39 and 40 as described above. The The imaging system including the objective lens 44, the magnification changing lens 35, the CCD camera 36, and the like is the same as the conventional structure, and thus detailed description thereof is omitted.

  Next, the control system (control means) of the culture observation system S in the present embodiment will be described with reference to the electric block diagram of FIG. As described above, the culture observation system S performs imaging of the incubator 2 that constitutes the culture environment in the culture chamber 13 and the cells (culture) that are cultured in the incubator 2 (in the culture chamber 13). The culture observation apparatus 1 provided with the apparatus 3 and the computer 5 connected to the culture observation apparatus 1 through a communication line. The culture observation apparatus 1 includes an incubator-side control device C1 and an imaging device-side control device C2.

An in-chamber temperature sensor 26, a CO ₂ gas concentration sensor 27, an O ₂ gas concentration sensor 28, a stage temperature sensor 46, and a control panel 16 are connected to the input side of the incubator-side control device C1, and the in-chamber temperature sensor 26 is connected to the output side. A heater 19, a door heater 18, a stage heater 41, a CO ₂ gas electromagnetic valve 24, an O ₂ gas electromagnetic valve 25, and a blower motor 32 are connected. Further, a communication line 50 for executing data communication with the computer 5 is connected to the incubator-side control device C1. Further, the incubator-side control device C1 has a built-in memory 53, and based on the output from the computer 5 and the output based on the operation of the control panel 16, each set value can be stored in the memory 53. Based on this, the temperature of the culture chamber 13, the gas concentration, and the temperature of the table 37 are controlled. Further, each temperature data and gas concentration data are output to the computer 5 by the incubator side control device C1.

  A lens 35, a CCD camera 36, stage motors 39 and 40, a light source 47, and the like constituting the imaging device 3 are connected to the output side of the imaging device side control device C2. Further, a communication line 51 (consisting of an image capturing communication line, a control communication line, etc.) for executing data communication with the computer 5 is connected to the image pickup apparatus side control apparatus C2. Thus, based on the output from the computer 5, control of coordinate movement to be observed, irradiation ON / OFF by the light source 27, brightness adjustment, image capture by the CCD camera 36, etc. is performed. Further, the coordinate position data to be observed and the luminance data of the light source are output to the computer 5 by the imaging device side control device C2.

  The computer 5 is provided with an application program (with a built-in memory 9 as a storage means) that enables the culture observation apparatus 1 to be controlled, to take in various data, and to reference and store the data. Various data and acquired images (microscope images) can be displayed on the display screen. Various settings can be executed by performing numerical input and determination operation according to the screen displayed on the display 4. A mouse 8 is connected to the computer 5 as an operation means.

Next, operation | movement of the culture observation system S of a present Example is demonstrated. By operating the control panel 16 or the computer 5, the culture conditions in the culture chamber 13, in this embodiment, the internal temperature, the internal CO ₂ gas concentration, and the O ₂ gas concentration are set. Here, the inside temperature is + 37.0 ° C., the CO ₂ gas concentration is 5.0%, and the O ₂ gas concentration is 5.0%.

  Based on this, the incubator-side control device C1 performs energization control of the in-compartment heater 19, the door heater 18, and the stage heater 41, and maintains the temperatures of the culture chamber 13 and the table 37 at the set temperature of + 37.0 ° C. Specifically, when the internal temperature sensor 26 decreases to + 36.5 ° C., the internal heater 19 and the door heater 18 are energized, and when the internal temperature sensor 26 increases to + 37.5 ° C., the internal heater 19 and the door heater 18 are energized. Is de-energized. Further, when the stage temperature sensor 46 is lowered to + 36.5 ° C., the stage heater 41 is energized, and when it is raised to + 37.5 ° C., the stage heater 41 is de-energized.

  Thus, the internal heater 19 and the door heater 18 are temperature-controlled independently of the stage heater 41. In this case, in this embodiment, the incubator-side control device C1 controls the temperature of the table 37 to be equal to or lower than the temperature in the culture chamber 13 and suitable for culturing cells as a culture. That is, the control device C1 sets the temperature of the table 37 to be the same as or lower than the temperature in the culture chamber 13 and is suitable for cell culture (in the present embodiment, +36. The temperature is controlled to be higher than 5 ° C.

  As a result, the air temperature in the culture chamber 13 can always be equal to or higher than the temperature of the table 37. Therefore, the temperature above the sample container 42 installed in the holding hole 43 of the table 37 (approximate to the air temperature in the culture chamber 13) is lower than the temperature below the sample container 42 contacting the table 37 (approximate to the temperature of the table 37). By lowering, it is possible to avoid the disadvantage that condensation occurs on the inner surface of the sample container 42.

Furthermore, the incubator side controller C1, based on the gas concentration in the cultivation room 13 detected by the CO ₂ gas concentration sensor 27 and the O ₂ gas concentration sensor 28 (CO ₂ gas concentration, O ₂ gas concentration), CO ₂ The gas solenoid valve 24, the O ₂ gas solenoid valve 25, and the exhaust valve 30 are controlled to open and close, and the inside of the culture chamber 13 is set to a set gas concentration (in this embodiment, the CO ₂ gas concentration is 5.0% and the O ₂ gas concentration is 5.0%).

On the other hand, since the humidification dish 55 as described in detail above is provided in the culture chamber 13, the humidifying water stored in the humidification dish 55 evaporates due to the air temperature in the culture chamber 13, and the culture is performed. Humidification of the chamber 13 is performed. In addition, since the gas introduction port 30 is connected to the air introduction port 58 of the humidifying tray 55 via the gas pipe 54, O ₂ gas or N ₂ gas introduced into the culture chamber 13 (in this embodiment) Since the set gas concentration of O ₂ is 5.0%, N ₂ gas) is once discharged into the water for humidification stored in the main body 56 by the air introduction port 58 of the humidifying dish 55 ( After being bubbled), it is introduced into the culture chamber 13 through a window 61 or the like formed in the lid 60 of the humidifying dish 55. Therefore, the inside of the culture chamber 13 is also efficiently humidified by the humidified N ₂ gas (or O ₂ gas) after being bubbled with the humidifying water in the humidifying dish 55.

  At this time, the humidity in the culture chamber 13 is preferably a humidity suitable for culture, for example, 90% or more. However, when the humidification proceeds, for example, 95% or more, dew condensation occurs in the culture chamber 13. Occurrence becomes remarkable.

  Here, the amount of evaporation from the humidifying water in the humidifying tray 55 varies depending on the area of the water surface exposed in the culture chamber 13. Therefore, when condensation on the inner wall surface of the culture chamber 13 or condensation on the sample container 42 occurs, the movable closing member 62 constituting the humidifying dish 55 is slid rearward to open the window 61 formed on the lid 60. Is adjusted to limit the open area of the water, that is, the exposed area of the humidifying water stored in the main body 56 to the culture chamber 13 (shown by a dotted line in FIG. 7).

  Thereby, the water surface of the humidifying water stored in the humidifying tray 55 is formed by the closing surface 57 formed at the lower edge of the window 61 and the upper surface 62A and the closing surface 62B of the movable closing member 62 that slides backward. The area of the water surface exposed in the culture chamber 13 is reduced.

  Therefore, the exposure area of the humidifying water into the culture chamber 13 is narrowed only to the projection surface of the window 61 and the area surrounded by the closed surfaces 57 and 62B. The amount of water used for evaporation is limited, and the degree of humidification is suppressed. Thereby, for example, the humidity in the culture chamber 13 is adjusted from 95% or more where condensation is likely to occur to a humidity suitable for cell culture, for example, about 91% or 92%. It becomes possible.

  Further, the water for humidification is evaporated from the water surface exposed in the enclosed culture chamber 13 at the closed surfaces 57 and 62B. , 62B and the bottom surface of the main body 56, water can be passed through, so that appropriate water supply is performed to the space portion.

  Furthermore, since the movable closing member 62 can be adjusted to open and close by arbitrarily sliding along the edge of the window 61, the movable closing member 62 is arbitrarily exposed in the culture chamber 13 with a simple structure. It is possible to finely adjust the area of the water surface. Thereby, it is aimed compared with the case where the water surface exposed in the culture chamber 13 is completely blocked by the lid 60 or the case where the water in the humidifying dish 55 is completely exposed in the culture chamber 13. It becomes easy to adjust the inside of the culture chamber 13 to the humidity.

  This makes it possible to easily adjust the humidity and maintain high humidity suitable for cell culture while suppressing the occurrence of condensation. Therefore, it is possible to avoid the dew condensation that occurs in the sample container 42 installed on the table 37. Therefore, the dew condensation avoids the inconvenience that the dew condensation interferes with the imaging of cells performed through the sample container 42. be able to. This makes it possible to appropriately observe the cells placed on the table 37.

  In addition, the movable closing member 62 overlaps any one of the plurality of engagement holes 64 formed in the upper surface 62A with the engagement hole 63 formed in the lid 60, and is screwed with a screw to obtain a predetermined It becomes possible to fix the exposed area. Thereby, by selecting the engagement hole 64, it becomes easy to adjust the exposed area of the water surface, it is possible to improve convenience, and avoid the inconvenience that the movable blocking member 62 easily moves. It becomes possible to do.

Furthermore, in this embodiment, the O ₂ gas or N ₂ gas introduced into the culture chamber 13 is introduced into the culture chamber 13 via the humidifying water in the humidifying dish 55, but the air introduction port 58 is provided outside the window 61, so that the water splash caused by the bubbling can be prevented from coming out of the humidifying pan 55 by the lid 60 and the closing surfaces 57 and 62B. It becomes.

Therefore, when the introduced O ₂ gas or N ₂ gas is humidified and introduced into the culture chamber 13, it is possible to avoid the inconvenience that the culture chamber 13 is immersed in water due to water splashing caused by bubbling. It becomes possible. This makes it possible to realize comfortable cell culture while enabling fine adjustment of humidity.

  The evaporation amount from the water surface of the humidifying water exposed in the culture chamber 13 can also be adjusted by opening and closing (detaching) the lid 60 itself constituting the humidifying dish 55. Therefore, when the humidity in the culture chamber 13 is increased at an early stage, the lid 60 is removed, the exposed area of the humidifying water stored in the main body 56 to the culture chamber 13 is increased, and the increase in evaporation amount is promoted. You can go.

  Moreover, although the water for humidification stored in the humidification dish 55 decreases by evaporation, it is preferable to add or replace in about one week to one month.

  With this configuration, the culture chamber 13 can be set to the temperature, gas concentration, and appropriate humidity set as described above. Therefore, by installing each sample container 42 containing cells or the like in each holding hole 43 of the table 37 provided in the culture chamber 13, the cells can be cultured according to the culture conditions. Note that the cells and the like placed on the table 37 can be observed by the imaging device 3. In the present embodiment, since a plurality of shelf plates 7 are installed in the culture chamber 13, the same culture conditions can be obtained by placing a container containing a culture (cells, etc.) on the shelf plate 7. This makes it possible to culture the cells.

It is a partial perspective front view of the culture observation apparatus which comprises the culture observation system to which this invention is applied. It is a side view of the culture observation apparatus of FIG. It is a vertical side view of the culture observation apparatus of FIG. It is a cross-sectional top view of the culture observation apparatus of FIG. It is an electric block diagram of the control apparatus which comprises a culture observation system. It is a vertical side view of a humidifying tray. It is a top view of the humidification dish of FIG. It is a vertical front view of the humidifying tray of FIG. It is a vertical side view of a lid. FIG. 10 is a longitudinal front view of the lid of FIG. 9.

S culture observation system 1 culture observation apparatus 2 incubator (culture means)
7 shelf (shelf)
10 opening 12 main body 13 culture chamber 15 insulating door 21 O ₂ · N ₂ gas inlet 22 N ₂ gas electromagnetic valve 23 CO ₂ gas supply port 24 CO ₂ gas electromagnetic valve 25 O ₂ gas electromagnetic valve 27 CO ₂ gas concentration sensor 28 O ₂ gas concentration sensor 37 Table 42 Sample container 54 Gas piping 55 Humidifying pan 56 Main body 57 Blocking surface 58 Air introduction port 60 Lid 61 Window 62 Movable closing member 62A Upper surface 62B Closing surface 63 Engagement hole

Claims (3)

In a humidifying dish for an incubator that is configured with a member having good heat conduction and is provided in a culture chamber of an incubator to store water for humidification,
A container-like main body, and a lid provided on the upper surface opening of the main body,
The lid includes a window for exposing the humidifying water to the culture chamber, a movable blocking member for adjusting the area of the water surface exposed in the window, and a position in the opening direction of the movable blocking member. A first closing part formed on the lower edge of the window side excluding the side to be hung and hanging down toward the bottom surface of the humidifying dish,
The movable closing member has an upper surface portion that slides on the upper surface of the lid, and a second closing portion that hangs from the end of the upper surface portion toward the bottom surface of the humidifying dish,
For the incubator, the area of the water surface exposed to the culture chamber surrounded by the first and second blocking portions can be adjusted by sliding the upper surface portion of the movable blocking member . Humidifying dish. The humidifying dish for incubators according to claim 1, wherein the lid has a gas introduction part located outside the window . An incubator comprising the humidifying dish for an incubator according to claim 1 or 2 in a culture chamber, wherein the culture chamber has a gas, temperature and humidity environment suitable for culture of the culture.

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