JP2020031545A - Culture device for microscopic observation - Google Patents

Abstract

To provide a culture device capable of suppressing as possible, stimulation to cells by works such as perfusion, exchange of a culture medium, administration of a liquid medicine, and mixing, especially, devising a configuration and shape of a feeding/discharge part for feeding and discharging a liquid to the culture container.SOLUTION: A perfusion IN pipe 25 and a liquid medicine IN pipe 31 as supply exclusive end parts are configured so that, tongue-shaped parts 25b, 31b where are formed by making lower end parts of the pipes into a half split shape, a supply liquid, for example, a liquid culture medium B or liquid medicine flows down through the tongue-shaped parts 25b, 31b, and is guided into a liquid level B1 of the liquid culture medium B in a dish 41. The liquid culture medium B or the like is supplied by discharge by a tube pump, however, a pulsation in discharging is eliminated in supplying. In addition, even an air is included in the tube, there is a sufficient distance between the base ends 25b, 31b and the liquid level B1, so that, the air is released to an upward before reaching a liquid level B1 in the dish 41. Therefore, bubbling is inhibited.SELECTED DRAWING: Figure 11

Description

  The present invention provides a microscopic observation including a culture vessel installed on a microscope stage, a supply / discharge unit for supplying and discharging a liquid to / from the culture vessel, and a controller capable of controlling the supply / discharge unit by a program operation. The present invention relates to a culturing device for use.

In the culture apparatus for microscopic observation disclosed in Patent Document 1, a culture vessel is set on a microscope stage for use.
Such devices are configured to maintain the culture environment, such as temperature and humidity, and perform operations such as perfusion, medium exchange, administration of chemicals, and mixing, either by the operator completely manually, Alternatively, manual operation is performed by pressing a button or the like.

Thus, it is difficult to standardize a completely manual operation due to a difference in the skill of the operator. The manual operation overcomes some of the disadvantages, but it also takes time to get the work done.
Recently, cells and the like have been continuously observed with a microscope at predetermined time intervals for a long time. Become.
On the other hand, if an operation command is issued from the controller by a program operation, the above-mentioned operation is completely separated from the operator's hand, but a skilled operator is required to perform delicate work to minimize damage to cells and the like. He is also careful and loses his attention.

JP 2008-259430A

  The present invention has been made in view of the above-described conventional problems, and in order to minimize the stimulation of cells, such as perfusion by a program operation, exchange of culture medium, administration of a drug solution, mixing, etc. It is an object of the present invention to provide a new and useful culture apparatus for microscopic observation, which devises a configuration of a supply / discharge unit for supplying and discharging a liquid to / from a culture container.

  The present invention has been made to solve the above problems, and the invention of claim 1 is a culture vessel installed on a microscope stage, and a supply and discharge unit for supplying and discharging a liquid to and from the culture vessel. In a culture apparatus for microscopic observation, comprising a controller capable of controlling the supply / discharge unit by a program operation, a supply-only end of the supply / discharge unit is suspended in the culture container, and a lower end side is half-split. Wherein the liquid is supplied along the tongue-shaped portion, flows down the tongue-shaped portion, and is guided to the liquid surface of the culture vessel when the liquid is supplied. Device.

  According to a second aspect of the present invention, in the culture apparatus for microscopic observation according to the first aspect, the discharge-dedicated end of the supply / discharge unit is formed of a pipe that is suspended in the culture vessel and has a horizontal hole, The lateral hole is disposed at an upper limit position of the liquid level, the supply / discharge end of the supply / discharge unit is suspended in the culture vessel, and is formed of a pipe having a lateral hole formed therein. The lateral hole is a culture device for microscopic observation, which is arranged at the lower limit position of the liquid level.

  According to a third aspect of the present invention, in the culture device for microscopic observation according to the second aspect, the discharge-dedicated end of the supply / discharge unit and the supply / discharge end are suspended close to each other, and the horizontal holes of each other are outward. A culture apparatus for microscopic observation, characterized in that the culture apparatus is arranged so as to face.

  According to a fourth aspect of the present invention, in the culture apparatus for microscopic observation according to the second or third aspect, the lateral hole of the supply / drain end is spirally formed along a side surface surrounding the liquid put in the culture container at the time of supplying the liquid. A culture apparatus for microscopic observation characterized in that it can be set at a position where the liquid flow is generated.

  The invention according to claim 5 is the culture device for microscopic observation according to claim 4, wherein the culture container is a dish having a circular cross section.

  According to a sixth aspect of the present invention, in the culture apparatus for microscopic observation according to the fourth or fifth aspect, the insert is detachable in the culture vessel, and the direction of the side hole of the supply / discharge end can be changed. This is a culture device for microscopic observation.

  According to a seventh aspect of the present invention, in the culture device for microscopic observation according to the sixth aspect, the holding portions of the supply / discharge end are formed at a plurality of positions, and the holding position of the supply / discharge end is changed. A culture apparatus for microscopic observation, characterized in that the direction of the horizontal hole can be changed between when an insert is used and when the insert is not used.

  According to an eighth aspect of the present invention, in the culture apparatus for microscopic observation according to any one of the first to seventh aspects, the supply / discharge unit supplies and discharges the liquid to / from the culture container by a tube pump. It is a culture device for use.

  According to the culture device for microscopic observation of the present invention, the configuration of the supply / discharge unit for supplying and discharging the liquid to / from the culture container is characterized by being operated by a program operation without the hand of a skilled worker. Even in this case, operations such as perfusion, medium exchange, administration of a drug solution, and mixing can be performed with high reliability.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an overall perspective view of a culture device for microscopic observation according to an embodiment of the present invention. FIG. 2 is an explanatory diagram of an operation of a pinch valve provided on a control side in FIG. 1. FIG. 2 is an exploded perspective view of an incubator including the incubation container of FIG. 1. FIG. 4 is a perspective view of the culture container of FIG. 3. FIG. 5 is a diagram schematically illustrating a positional relationship between a dish and various pipes without an insert in FIG. 4. FIG. 6 is an explanatory diagram of shapes of various pipes in FIG. 5. It is explanatory drawing of the arrangement | positioning state of the pipe in the dish of FIG. FIG. 5 is a diagram schematically illustrating a positional relationship between a dish and various pipes in a case where an insert shown in FIG. 4 is provided. It is explanatory drawing of the arrangement state of the pipe in the dish of FIG. FIG. 5 is an explanatory view of a pipe holding groove of the culture vessel of FIG. 4. It is an image figure of continuous perfusion. It is an image figure of exchange of a culture medium. It is an image figure of administration of a medical solution. FIG. 9 is an image diagram of mixing of a chemical solution (without insert). FIG. 7 is an image diagram of mixing of a chemical solution (in a case where an insert is provided).

A culture apparatus 1 for microscopic observation according to an embodiment of the present invention will be described below with reference to the drawings.
In addition, in order to maintain the culture environment, other maintenance mechanisms such as humidity and temperature are also provided.However, since the features of the present invention are in the supply / discharge unit, illustration and description of other parts are omitted as appropriate. .

In the culture apparatus 1 for microscopic observation, as shown in FIG. 1, a controller 3 is used. The controller 3 includes a microcomputer, and control means is realized by the program operation.
As shown in FIG. 1, tube pumps 7 and 9 are attached to the outer surface of a box-shaped housing 5 of the controller 3. In these tube pumps 7 and 9, a flexible tube is arranged between a pressing member and an outer member having a circular inner periphery, and the pressing member is eccentrically moved to sequentially press the tubes to perform a pumping operation. This is used for supply and discharge of a liquid medium because the structure is simple and liquid can be sent without contaminating the inside of the tube.

These tube pumps 7, 9 are driven independently of each other. These driving motors (not shown) are both installed in the housing 5. The motor of the tube pump 7 can rotate forward and backward.
One of the flexible tubes disposed in the tube pump 7 is branched on one side in the flow direction, and one is a perfusion OUT (upper limit) tube 11 and the other is a perfusion OUT (lower limit) tube 15. A pinch valve 19 is also attached to the same surface side of the housing 5, and a perfusion OUT (upper limit) tube 11 and a perfusion OUT (lower limit) tube 15 are attached to the pinch valve 19. As shown in FIG. 2, the shaft 21 of the pinch valve 19 is interposed between the tubes 11 and 15, and when not in operation, the perfusion OUT (upper limit) tube 11 is crushed to make it inconsistent while the perfusion OUT ( The lower limit) tube 15 is opened, and during operation, the perfusion OUT (upper limit) tube 11 is opened, while the perfusion OUT (lower limit) tube 15 is crushed so as to be disconnected.
The other side in the flow direction is connected to a waste liquid system (not shown).

The flexible tube arranged in the tube pump 9 is a perfusion IN tube 23. One side in the flow direction of the perfusion IN tube 23 is connected to a supply system (not shown) for the liquid medium B.
A dedicated air pump connection 26 is further provided on the outer surface of the housing 5 for administering a chemical solution. An air pump (not shown) is provided inside the housing 5, and a pumping function works through the air pump connection portion 26. To the air pump connecting portion 26, a tubular chemical solution setting portion 27 is detachably connected. FIG. 1 shows a state where one end side is connected to the air pump connection part 26 after the chemical liquid is set inside the chemical liquid setting part 27. One end of the flexible tube in the flowing direction of the flexible tube is detachably connected to the other end of the chemical solution setting section 27, and this is a chemical solution IN tube 29.

The control means realized by the program operation of the controller 3 controls the operations of the tube pumps 7 and 9, the pinch valve 19, and the air pump.
One end of the perfusion OUT (upper limit) tube 11 and the perfusion direction of the perfusion OUT (lower limit) tube 15 and the other end of the perfusion IN tube 23 and the chemical solution IN tube 29 in the flow direction are connected to rigid pipes, respectively. These are a perfusion OUT (upper limit) pipe 13, a perfusion OUT (lower limit) pipe 17, a perfusion IN pipe 25, and a chemical IN pipe 31.
These pipes 13, 17, 25, 31 constitute a supply / discharge unit.

As shown in FIG. 3, the main body of the incubator 33 includes a housing unit 35 and a lid 37 that closes an upper opening of the housing unit 35. Is a space for accommodating the culture container 39 for accommodating cells and tissues to be observed together with the liquid medium B.
The container body of the culture container 39 is a transparent dish 41. The dish 41 has a shallow bottomed cylindrical shape with a circular bottom surface. An insert 43 is inserted into the dish 41 as necessary to change the capacity. A diamond-shaped hole is formed at the center of the insert 43 so that cells or the like can be inserted into the center of the dish 41 with or without the use of the insert 43. When the insert 43 is used, the upper surface of the opening of the dish 41 is suppressed by the insert holder 45 and then closed by the fixing lid 47. When the insert 43 is not used, the upper surface of the opening of the dish 41 is directly closed by the fixing lid 47. The central part of the fixed lid 47 is opened in a circular shape to form an opening 49, which can be opened and closed by the liquid medicine administration lid 57. The medical solution administration lid 57 also serves as a light transmitting part for microscope observation.

As shown in the enlarged view of FIG. 4, the above-mentioned pipes 13, 17, 25, 31 are positioned and fixed to the fixing lid 47.
The fixed lid 47 is notched horizontally long along the edge of the opening, and a portion surrounded by the edge of the notch and the edge of the drug administration lid 57 is a pipe insertion hole 51. A pipe holding groove 53 extends linearly from the pipe insertion hole 51 to the outer peripheral edge of the fixed lid 47. The pipe holding groove 53 is concave and opens upward.
Two sets of the pipe insertion hole 51 and the two pipe holding grooves 53 are provided in total, and two sets are provided on each diametrically opposite side of one of the fixed lids 47. The two pipe holding grooves 53, 53 forming a pair on the same side, extend parallel to each other.
Further, in addition to the pipe holding groove 53, a pipe holding groove 55 is connected to the one pipe insertion hole 51 in a similar configuration. The pipe holding groove 55 and the pipe holding groove 53 are substantially orthogonal.

The dish 41 is attached to the attachment 59. The attachment 59 is provided with a fitting hole 61 and a holding piece 63. The bottom of the dish 41 is fitted into the fitting hole 61 and is elastically suppressed by the holding piece 63, so that the dish 41 is attached to the attachment 59. Fixed. The attachment 59 is attached to the bottom opening of the storage unit 35 of the incubator 33.
When using the culture apparatus 1 for microscopic observation, as shown in FIG. 4, a fixed lid 47 is attached to the dish 41 to form a culture vessel 39, and is stored in the incubator 33. Installed in

As shown in detail in FIGS. 5 to 7, each of the pipes 13, 17, 25, and 31 is bent vertically in the middle to form an L-shape, and the horizontal portions are pipe holding grooves 53, 53, .. Are placed and held, and the vertical part is suspended in the dish 41 through the pipe insertion hole 51, and forms the suspended parts 13 a, 17 a, 25 a, and 31 a, respectively.
The lower ends of the hanging portions 13a, 17a of the pipes 13, 17 are both closed.
As shown in FIG. 6, a horizontal hole 13 p having a circular cross section is formed in a hanging portion 13 a of the perfusion OUT (upper limit) pipe 13, and a similar lateral hole is formed in a hanging portion 17 a of the perfusion OUT (lower limit) pipe 17. Although the hole 17p is formed, the hole 17p is formed at a position different in distance from the lower end closing portion, and the height position H1 of the horizontal hole 13p is higher than the height position H2 of the horizontal hole 17p.
In this embodiment, the dish 41 is of a commercially available 35 mm size, and accordingly, H1 is set to 4 mm and H2 is set to 1.2 mm.

The lower end of the hanging part 25a of the perfusion IN pipe 25 is divided in half and connected in a tongue shape. A base end 25c of the tongue-like portion 25b is an opening of the pipe. The height of the base end 25c is higher than that of the horizontal hole 13p of the perfusion OUT (upper limit) pipe 13. The base end 25c is designed to be higher than the liquid level of the medium placed in the dish 41.
The chemical IN pipe 31 is formed in the same shape and the same size as the perfusion IN pipe 25.

FIGS. 5 to 7 show a state where the pipes 13, 17, 25 and 31 are arranged on the dish 41 without the insert.
From one of the pipe insertion holes 51, a perfusion OUT (upper limit) pipe 13 and a perfusion OUT (lower limit) pipe 17 are suspended. The hanging portions 13a and 17a are hung parallel to each other, and the lower ends of the hanging portions 13a and 17a are in contact with the bottom upper surface of the dish 41. The horizontal holes 13p and 17p face outward so as not to face each other. In FIG. 7, the horizontal holes 13p and 17p are oriented in the opposite direction (that is, 180 °), but are not limited to this, and may be about 90 °.
When the hanging portions 13a, 17a of the pipes 13, 17 are close to each other, they are susceptible to the effect of surface tension. In this way, the horizontal holes 13p, 17p are not opposed to each other, and in particular, in this embodiment, In the case of (1), since they face in the opposite direction, there is no adverse effect on the leveling accuracy of the liquid surface and the mixing operation.

  From the other pipe insertion hole 51, a perfusion IN pipe 25 and a chemical IN pipe 31 are suspended. The hanging portions 25a and 31a are hung in parallel with each other, and the lower ends thereof are in contact with the bottom upper surface of the dish 41. The concave surfaces of the tongue-like portions 25b and 31b are both facing the center of the dish 41. Therefore, as shown by the arrow in FIG. 7, the flowing liquid has a gentle flow toward the perfusion OUT (upper limit) pipe 13 and the perfusion OUT (lower limit) pipe 17.

8 and 9 show a state where the pipes 13, 17, 25 and 31 are arranged in the dish 41 in the case where the insert is present.
When the insert 43 is placed in the dish 41, two acute-angled portions 43a, 43a of the diamond-shaped hole edge of the insert 43 are approaching the inner surface of the dish 41, and the pipes 13, 17, 25, 31 is hung down. As described above, in consideration of the case where the insert 43 is used, the pipes 13 and 17 and the pipes 25 and 31 hang down in a state of being close to each other, but as described above, the pipes 13 and 17 Has been devised so as not to exert an adverse effect of surface tension.
Also, as shown in FIGS. 4 and 5, when there is no insert, all the pipes are held in the pipe holding groove 53. In this case, the perfusion OUT (lower limit) pipe 17 is held in the pipe holding groove 55. Thus, the position of the side hole 17p is changed.

  As shown in FIG. 10, the horizontal portion of the perfusion OUT (lower limit) pipe 17 is selected and held by any of the pipe holding grooves 53 and 55, so that the orientation of the horizontal hole 17p of the hanging portion 17a as described above. Can be changed.

  The supply / discharge unit is configured as described above, and the operation of the tube pumps 7 and 9, the pinch valve 19, and the air pump is performed by the operation command from the controller 3 with respect to the cells and the like placed in the dish 41. Under control, the operations of continuous perfusion, medium exchange, administration of a drug solution, and mixing of drug solutions proceed.

(A) Continuous perfusion (FIG. 11)
The liquid medium B is supplied into the dish 41 from the perfusion IN pipe 25 by the operation of the tube pump 9.
The tube pump 9 generates pulsation due to its structure, and it is difficult to make the flow rate constant. However, since the tongue-like portion 25b of the perfusion IN pipe 25 enters the liquid medium B, the tube pump 9 is sent by the tube pump 9. The liquid medium B that has flowed down the tongue-shaped portion 25b flows down the liquid surface B1 of the liquid medium B in the dish 41, and is supplied in a state where the pulsation at the time of discharge has disappeared. Since the tongue-like portion 25b enters below the liquid level B1, the liquid medium B to be supplied does not drip when the liquid medium B is supplied.
Even if air (air) A is entrained in the perfusion IN pipe 25, there is a sufficient distance between the base end 25c and the liquid level B1, so that the air A reaches the liquid level B1 in the dish 41. Escape upward as indicated by the arrow. Therefore, occurrence of bubbling is prevented.

At the same time, the liquid medium B that is going to exceed the liquid level B1 is sucked and discharged through the perfusion OUT (upper limit) pipe 13 by the operation of the tube pump 7. When the liquid surface B1 of the liquid culture medium B is lower than the horizontal hole 13p, air (air) is sucked. When the liquid surface B1 of the liquid culture medium B covers the horizontal hole 13p, the liquid culture medium B is suctioned and discharged. Is done.
Since the suction flow rate is set to be slightly larger than the supply flow rate, the leveling by the lateral holes 13p has high accuracy.
Further, since the pipe 25 on the supply side and the pipe 13 on the suction side are substantially diagonal and are implemented at a low flow rate, the flow of the liquid is gentle without disturbing the liquid level B1.

(B) Change of medium (FIG. 12)
By the operation of the tube pump 7, the liquid is sucked and discharged from the dish 41 through the perfusion OUT (lower limit) pipe 17 until the liquid surface B1 of the liquid medium B reaches the lower limit position. When continuous perfusion has been performed immediately before, the liquid surface B1 of the liquid medium B is at the height of the horizontal hole 13p, so the discharge is continued until the liquid surface B1 drops to the height of the horizontal hole 17p. I do.
Thereafter, the liquid medium B is discharged and supplied into the dish 41 from the perfusion IN pipe 25 by the operation of the tube pump 9. In parallel with the supply, the liquid is discharged from the perfusion OUT (upper limit) pipe 13 by the operation of the tube pump 7, so that the liquid level B1 does not exceed the upper limit position. The same operation as in the above-mentioned “(A) Continuous perfusion” is performed at the time of discharge supply by the perfusion IN pipe 25 and suction and discharge by the perfusion OUT (upper limit) pipe 13.

(C) Administration of drug solution (FIG. 13)
The chemical Y is discharged from the chemical IN pipe 31 into the dish 41 by the operation of the air pump. In parallel with the supply, the liquid is discharged from the perfusion OUT (upper limit) pipe 13 by the operation of the tube pump 7, so that the liquid level B1 does not exceed the upper limit position.
The chemical | medical solution IN pipe 31 acts similarly to the perfusion IN pipe 25 of "(A) Continuous perfusion". Since the air pump is used, air (air) is easily entrained. However, since the air (air) escapes as described above, bubbling is effectively prevented.

(D) Mixing of chemicals (FIGS. 14 and 15)
The operation direction of the tube pump 7 is alternately switched between suction and discharge.
By the suction operation of the tube pump 7, the liquid Y is sucked and discharged together with the liquid medium B into the perfusion OUT (lower limit) pipe 17 and stays in the tube, and then, by the discharge operation of the tube pump 7, the perfusion OUT (lower limit) is performed. The liquid Y is discharged from the pipe 17 and returned into the dish 41 together with the liquid medium B.
This suction / discharge and discharge / supply are repeated.

  Since the flow rate is set large according to the purpose of mixing, a liquid flow is generated in both supply and discharge. When the lower end of the pipe is opened and used as a lower hole separated from the upper surface of the dish 41, the liquid of the liquid culture medium B coming out of the lower hole is supplied to the lower upper surface of the adjacent dish 41 particularly at the time of supply. Although the flow collides and the momentum is rapidly scraped off, in this embodiment, since the lateral hole 17p is used, the momentum is not shaved off by the collision with the bottom upper surface of the dish 41. .

As shown in the left diagram of FIG. 14, when there is no insert, the direction of discharge from the lateral hole 17p is as indicated by the arrow, and the liquid flow of the liquid medium B collides with the opposing inner surface 41a of the dish 41. Thereafter, a vortex flows along the entire inner surface, so that the flow of the liquid is not reduced and the mixing is promoted. Further, damage to cells S and the like placed at the center of the dish 41 is minimized.
Similarly, when the liquid medium B is sucked, the liquid flow becomes a vortex.
Therefore, as shown in the right diagram of FIG. 14, the vortex in the opposite direction is generated alternately by the discharge and the suction, and the mixing is promoted by this interaction.

When the direction of the lateral hole 17p is arranged in the same manner as the case without the insert in the case with the insert, the liquid flow of the liquid culture medium B discharged from the lateral hole 17p collides with the inner surface of the very close insert 43. Therefore, while the liquid medium B spouts up, the momentum is rapidly shaved. However, by holding the perfusion OUT (lower limit) pipe 17 in the pipe holding groove 55, the horizontal hole 17p comes to the position shown in FIG. That is, as shown in FIG. 15, the discharge direction from the horizontal hole 17p can be changed as shown by the arrow. Although the flow rate is reduced in consideration of the mixing in the insert 43 having a limited capacity, a liquid flow having sufficient force for mixing the liquid medium B supplied thereto is secured in accordance with the capacity of the insert 43. .
Thus, it is possible to cope with the case where the insert 43 is used. Note that the horizontal hole 17p is not at the position directly opposite to the horizontal hole 13p, but is separated by 90 °, so that the influence of surface tension can be suppressed.

  As described above, in the operations (A), (B), (C), and (D), the shape features and arrangements of the pipes 13, 17, 25, and 31 forming the ends of the supply / discharge unit are utilized. Thus, each operation is performed with high reliability while minimizing damage to the cells S and the like.

Although the embodiment of the present invention has been described in detail above, the specific configuration is not limited to this embodiment, and even if there is a change in the design without departing from the gist of the present invention, the invention is not limited to the embodiment. included.
For example, the culture vessel is not limited to a dish, but includes a well plate and the like. However, when a dish having a circular cross section is used, it is a preferable culture vessel because the liquid flow can be easily guided so as to draw a beautiful vortex during the mixing operation, taking advantage of its shape. I have.

DESCRIPTION OF SYMBOLS 1 ... Microscope observation culture apparatus 3 ... Controller 5 ... Housing 7, 9 ... Tube pump 11 ... Perfusion OUT (upper limit) tube 13 ... Perfusion OUT (upper limit) pipe 13a ... Hanging part 13p ... Horizontal hole 15 ... Perfusion OUT (lower limit) ) Tube 17 ... Perfusion OUT (lower limit) pipe 17a ... Hanging part 17p ... Side hole 19 ... Pinch valve 21 ... Shaft 23 ... Perfusion IN tube 25 ... Perfusion IN pipe 25a ... Hanging part 25b ... Tongue piece 25c ... Base end 26 ... Air pump connection part 27 ... Chemical liquid setting part 29 ... Chemical liquid IN tube 31 ... Chemical liquid IN pulp 31a ... Hanging part 31b ... Tongue-shaped part 31c ... Basic end 33 ... Incubator 35 ... Container unit 37 ... Cover 39 ... Culture container 41 ... Dish 41a ... Inner surface (of the dish) 43 ... Insert 43a ... Acute angle part (of the insert) 45 Insert holder 47 ... Fixed lid 49 ... Opening 51 ... Pipe insertion hole 53 ... Pipe holding groove 55 ... Pipe holding groove 57 ... Chemical liquid administration lid 59 ... Attachment 61 ... Fit hole 63 ... Pressing piece B ... Liquid medium B1 ... (Liquid Liquid level of culture medium Y ... Drug solution S ... Cells etc. H1, H2 ... Hole height

Claims (8)

A culture vessel installed on a microscope stage, a supply / discharge unit for supplying and discharging a liquid to / from the culture vessel, and a culture apparatus for microscopic observation comprising a controller capable of controlling the supply / discharge unit by a program operation. ,
The supply-only end of the supply / drain section is suspended in the culture vessel, and is formed of a pipe connected in a tongue-like shape with a lower end half-split,
A culture apparatus for microscopic observation, wherein, at the time of supplying a liquid, the liquid flows down the tongue-shaped portion and flows down to the liquid surface of the culture vessel. The culture device for microscopic observation according to claim 1,
The discharge-only end of the supply / discharge unit is suspended in the culture vessel, and is formed of a pipe formed with a horizontal hole, and the horizontal hole is disposed at an upper limit position of a liquid level,
The supply / discharge end of the supply / drain section is suspended in the culture vessel, and is formed of a pipe having a horizontal hole formed therein, and the horizontal hole is disposed at a lower limit position of a liquid level. Characteristic culture equipment for microscopic observation. The culture device for microscopic observation according to claim 2,
A culture for microscopic observation, characterized in that the discharge-only end of the supply / discharge unit and the supply / discharge end are suspended in close proximity, and the lateral holes are arranged facing outward. apparatus. The culture device for microscopic observation according to claim 2 or 3,
The horizontal hole in the supply / discharge end can be set at a position where a vortex liquid flow is generated along the side surface surrounding the liquid put in the culture vessel when the liquid is supplied, for microscopic observation. Culture device. The culture apparatus for microscopic observation according to claim 4,
A culture vessel for microscopic observation, wherein the culture vessel is a dish having a circular cross section. The culture device for microscopic observation according to claim 4 or 5,
A culture apparatus for microscopic observation, wherein an insert is detachable in a culture vessel, and a direction of a side hole of a supply / discharge end is changeable. The culture device for microscopic observation according to claim 6,
The holding portion of the supply / discharge end is formed at a plurality of positions, and by changing the holding position of the supply / discharge end, the direction of the horizontal hole is changed depending on whether the insert is used or not. A culture device for microscopic observation characterized by being changeable. The culture device for microscopic observation according to any one of claims 1 to 7,
A culture apparatus for microscopic observation, wherein a supply / discharge unit supplies and discharges a liquid to / from a culture container by a tube pump.

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