JP6201647B2 - Microscope observation culture equipment - Google Patents

Description

  The present invention relates to a culture apparatus for microscopic observation, and more particularly to a culture apparatus for microscopic observation suitable for observing living cells for a long period of time.

For example, in the field of cell research, the phenomenon is observed while culturing cells for a long time. In general, in order to culture cells for a long period of time, it is preferable to maintain a suitable environment for the cells. For example, as an environment for culturing common cell, temperature 37 degrees, it is that it is preferable to maintain humidity of 90%, the CO ₂ concentration of 5%.

  FIG. 5 is a configuration explanatory view showing an example of a conventional incubator for microscope observation (Patent Document 1). In FIG. 5, the petri dish 1 containing cells as observation materials is held inside a sealed incubator, a reservoir 2 is provided around the petri dish, and a heater 3 is disposed below the reservoir 2, thereby culturing. An example in which the humidity inside the container is kept in a high humidity state is disclosed.

JP 2003-107364 A

  However, as shown in FIG. 5, simply providing the reservoir 2 around the cell and heating from the lower surface cannot sufficiently promote evaporation, and it is difficult to maintain the humidity at a high humidity suitable for cell culture. is there.

  The present invention solves such a problem, and the object thereof is a culture for microscopic observation that can maintain a humidity environment suitable for cell culture in a microscopic observation culture apparatus that performs microscopic observation while culturing living cells. To implement the device.

In order to achieve such a problem, the invention according to claim 1 of the present invention is:
In a microscopic observation culture apparatus that performs microscopic observation while culturing living cells,
A top heater unit and a bottom heater unit are provided so as to sandwich the XY stage unit from above and below,
The XY stage unit is provided with a plate holder for holding a well plate containing a sample being cultured ,
The bottom heater unit is characterized in that a flow path including a heater holder and a metal plate is formed on one surface, and a heater is provided on the other surface .

The invention of claim 2 is the culture apparatus for microscopic observation according to claim 1,
The XY stage unit can be largely driven in at least one of XY directions so that the well plate can be attached and detached.

The invention of claim 3 is the culture apparatus for microscopic observation according to claim 1 or 2,
The plate holder is provided with at least one of a heating heater and a CO ₂ port for supplying CO ₂ gas from the outside.

  By these, the culture apparatus for microscope observation which can maintain the humidity environment suitable for cell culture is realizable.

It is a front view of one Example of this invention. It is AA sectional drawing of FIG. It is a perspective view which shows the internal structure of FIG. It is an exploded view of the bottom heater 6 of FIG. It is a configuration explanatory view showing an example of a conventional incubator for microscopic observation.

  Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. FIG. 1 is a front view of an embodiment of the present invention, and FIG. 2 is a cross-sectional view taken along line AA of FIG.

  A top heater unit 20 is provided at the top of the culture apparatus 10 according to the present invention, an XY stage unit 30 is provided at the bottom of the top heater unit 20, and a bottom heater unit 40 is provided at the bottom of the XY stage unit 30. ing.

  The top heater unit 20 includes a transparent heater panel 21, a holder 22 that holds the heater panel 21, and a fixing plate 23 that fixes these to a fixing portion (not shown). In the center of the heater panel 21, a hole 21a for adding a chemical solution is provided.

  The XY stage unit 30 is provided with a plate holder 52 for holding a well plate 51 in which a sample being cultured is accommodated. Further, the XY stage unit 30 has an X table 31 for driving the well plate 51 in the X direction and an Y table 32 for driving in the Y direction by an actuator (not shown), and the well plate 51 can be driven in the XY direction. .

  Further, the XY stage unit 30 is configured to be able to attach and detach the well plate 51 by largely driving in the Y direction to move the plate holder 52 to the outside from the position covered with the top heater unit 20. .

A heater (not shown) is affixed to the plate holder 52 so that it can be heated. The plate holder 52 is provided with a CO ₂ port (not shown) so that CO ₂ gas adjusted to 5%, for example, can be supplied from the outside.

  The plate holder 52 is provided with a seal rubber 53 so as to keep airtight with the heater panel 21.

  A seal rubber 33 is provided on the bottom surface of the Y table 32 and is configured to keep airtight with the bottom heater 40.

  FIG. 3 is an explanatory view in which the heater panel 21 and the well plate 51 are removed from FIGS. 1 and 2, and FIG. 4 is an exploded view of the bottom heater unit 40.

In FIG. 4, an opening hole h is provided at the center of the bottom heater unit 40, and an objective lens 60 is disposed. The objective lens 60 is configured to be driven in the optical axis direction by an actuator (not shown). Further, a bellows 61 is provided at the edge of the objective lens 60 and the opening of the bottom heater unit 40. When the objective lens 60 is raised, the environment such as the internal temperature, humidity, and CO ₂ is exposed to the outside. It is configured not to leak.

  Although details are omitted, an image of cells inside the well plate 51 photographed by the objective lens 60 is configured to be photographed by an optical system and a camera (not shown).

  The bottom heater unit 40 includes a heater holder 41 and heater frames 42 a and 42 b that fix and support both ends of the heater holder 41. A sheet-like heater 43 is affixed to the lower surface of the heater holder 41 so as to be heated. Further, the opening hole h is provided at the center of the heater holder 41 as described above, and the above-described bellows 61 is fixed.

  In the heater holder 41, the peripheral portion 41a of the opening hole h on the surface is processed to be slightly lower (for example, about 0.5 mm) than the outer peripheral portion 41b of the peripheral portion 41a so as to form a flow path.

  Further, a water supply hole 41c and a drain hole 41d are provided on the surface of the heater holder 41 so that water supply and drainage can be performed from a port on the bottom surface (not shown).

  A metal plate 45 is affixed to a peripheral portion 41 a formed on the surface of the heater holder 41 via a metal mesh 44 that is processed and formed so as to be fitted to the peripheral portion 41 a, and a flow path is formed by the heater holder 41 and the metal plate 45. Is formed. In the predetermined region 45a of the metal plate 45, for example, minute holes having a diameter of 0.5 mm are formed in a staggered manner with a pitch of 1 mm.

  In such a configuration, when the well plate 51 to which the observation sample is attached is set in the plate holder 52, the XY stage unit 30 is largely driven in the Y direction so that the plate holder 52 is covered with the top heater unit 20. The well plate 51 to which the observation sample is attached is set in the plate holder 52. Thereafter, the XY stage unit 30 is returned to the observation position.

  By moving the objective lens 60 in this state, the sample attached to the well plate 51 can be observed by an optical system and a camera (not shown). Here, illumination may be performed through the transparent heater panel 21 by a lamp (not shown) from above, or illumination may be performed through the inside of the objective lens 60 from the optical system side. Further, by driving the XY stage unit 30, a desired position inside the well plate 51 can be observed.

  Regarding the temperature, the heater is heated by energizing the heater 43 (not shown) attached to the transparent heater panel 21 and the plate holder 52 and the heater 43 of the bottom heater unit 40, and the inside of the well plate 51 is, for example, 37 by a thermocouple (not shown). Control to reach ℃.

As for CO ₂ , the CO ₂ concentration in the vicinity of the well plate 51 inside the culture apparatus 10 is adjusted by supplying a gas mixture whose CO ₂ concentration is adjusted to, for example, 5% from the outside through a CO ₂ port (not shown) of the plate holder 52. Can be 5%.

  As for humidity, pure water is supplied from the water supply hole 41 c of the bottom heater unit 40. This water enters a thin flow path formed by the heater holder 41 and the metal plate 45 by capillary action. Since the metal mesh 44 is disposed in this flow path, the flow path becomes narrower and the capillary phenomenon is enhanced, so that water is efficiently guided inside the flow path.

  In addition, the amount of water supplied from the water supply hole 41c and the amount of water discharged from the drain hole 41d are adjusted so that water does not overflow from a large number of minute holes provided in the metal plate 45. Here, since the diameter of the minute hole is as small as 0.5 mm, for example, surface tension works to prevent water from coming out of the minute hole, and water is more efficiently guided into the flow path.

  Further, since the water layer inside the flow path is very thin and has a very small volume, it is easily heated by the heater 43, and evaporation is actively promoted from a large number of minute holes provided in the metal plate 45. . For this reason, humidity improves efficiently. Further, the metal mesh 44 disposed inside the flow path also plays a role of transferring heat more efficiently.

  The heater 13 is set to 38.5 degrees, which is slightly higher than the internal temperature of 37 degrees. Evaporation of water is performed in a large number of minute holes provided in the metal plate 45. Since the distance between these minute holes and the heater 13 is short, the temperature of this part is heated to substantially the same temperature as the heater 13.

  Therefore, since the temperature of the evaporated saturated vapor is also higher than 37 ° C., convection occurs and the saturated vapor convects inside the culture apparatus, and an unsaturated gas convects in the vicinity of the metal plate 45. Evaporation is promoted. With these effects, it is possible to construct a humidity environment that is more efficient and higher than usual.

  Instead of the metal plate 45, a metal mesh that has been processed into a similar outer shape and subjected to water-repellent processing may be used.

  Moreover, not only a metal mesh but what has the same effect, such as steel wool and absorbent cotton, may be used.

  As described above, according to the present invention, it is possible to realize a culture apparatus for microscope observation that can maintain a humidity environment suitable for cell culture.

DESCRIPTION OF SYMBOLS 10 Culture apparatus 20 Top heater unit 30 XY stage unit 40 Bottom heater unit

Claims (3)

In a microscopic observation culture apparatus that performs microscopic observation while culturing living cells,
A top heater unit and a bottom heater unit are provided so as to sandwich the XY stage unit from above and below,
The XY stage unit is provided with a plate holder for holding a well plate containing a sample being cultured ,
A culture apparatus for microscopic observation, wherein the bottom heater unit has a flow path formed of a heater holder and a metal plate on one surface and a heater on the other surface .   The culture apparatus for microscope observation according to claim 1, wherein the XY stage unit can be largely driven in at least one of XY directions so that the well plate can be attached and detached. 3. The microscope holder according to claim 1, wherein the plate holder is provided with at least one of a heating heater and a CO ₂ port for supplying CO ₂ gas from the outside. Culture device.