JP2014095786A - Culture device for microscopic observation and attachment for container - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a culture device for microscopic observation capable of performing microscopic observation of a sample of a cell and the like in a culture solution at a high magnification and an attachment for a container used for the same.SOLUTION: Heat is transmitted to a culture solution downward from the top. An attachment 45 for a container is configured to include: a guide base part 47 fitted into a lower opening 49 of a container unit and having a flat bottom face; the opening 49 formed therein; a step part provided along a mouth edge of the opening 49 and made one-step higher than the surplus bottom face; and a pair of pressure welding parts 69, 79 facing to each other by nipping the opening 49 on the guide base part 47. A slide glass plate S of the culture container is made to enter from a lower side, and the step part of the slide glass plate S is made to abut against the step part, and the bottom face of the plate and the bottom face of the guide base part 47 are made approximately flat and pressure-welded by the pressuring welding parts 69, 79. An object lens can be brought fully closer even at a corner of the plate so that the observation can be made at high magnification.

Description

  The present invention relates to a microscopic observation culture apparatus used for microscopic observation of a sample such as a cell, and a container attachment used when a culture container is attached to the apparatus.

A typical example of this type of microscopic observation culture apparatus is shown in Patent Document 1.
As shown in FIG. 21, the microscope observation culture apparatus 201 is installed on a microscope stage and used, and the main body is composed of a storage unit 203 and a lid 205 that closes the upper opening. Has been. Usually, a water tank portion 20 7 is provided along the inner edge side in the storage unit 203, and the inside is a storage space for the culture vessel. The dish D, which is a culture container, is accommodated in the accommodation space in a state of being fitted into the container attachment 209 from above.
The culture solution placed in the dish D needs to be maintained at a constant level, for example, about 37 ° C., in order to maintain the sample cells and the like under conditions suitable for observation. Therefore, conventionally, the container attachment 209 has a heat generation or heat transfer action, and heat is transmitted from the dish D to the dish D to heat the culture medium therein. Further, the temperature sensor 211 is disposed on the lid 205 side, and the temperature is controlled based on the temperature information.

JP 2008-259430 A

Thus, recently, in order to perform microscopic observation at a high magnification, when an inverted microscope is used, the objective lens tends to be closer to the lower surface of the housing unit 203 than before, but as described above, When the attachment 209 is interposed, in the vicinity of the edge of the container attachment 209, the edge becomes an obstacle and the approach does not remain.
In addition, when the culture solution is to be heated from the container attachment 209 via the wall of the dish D, a difference in the distance from the container attachment 209 occurs in the dish D and a temperature gradient is generated. In particular, when the culture vessel is divided into a plurality of wells like a slide glass plate, convection of the culture solution cannot be expected, and the temperature gradient becomes larger.

  Furthermore, in order to detect the temperature of the culture solution, the most accurate detection is possible if the temperature detection part can be directly detected by immersing it in the culture solution. As described above, when observing with high accuracy that cares about temperature gradients, place the temperature detector at the center. Such detection is not realistic because the observation location is narrowed.

Therefore, an object of the present invention is to provide a culture apparatus for microscopic observation that solves the above-described various problems and enables efficient observation of a culture container at a high magnification.
Another object of the present invention is to provide a container attachment suitable for use as an accessory for the above-described culture apparatus for microscopic observation.

  The present invention solves the above-mentioned problems, and the invention according to claim 1 is an attachment for a container used for accommodating a container in a storage unit of a culture apparatus for microscopic observation, and is provided under the storage unit. A guide base portion having a flat bottom surface to be fitted into the opening portion, an opening portion formed in the guide base portion, and a step provided along the lip of the opening portion, which is one step higher than the remaining bottom surface. And a pair of pressure contact portions opposed to each other across the opening on the guide base portion, and at least one of the pressure contact portions is supported so as to be able to reciprocate in the proximity and separation direction with respect to the other pressure contact portion. A pressure contact portion supporting means; and a pressure contact portion fixing means for fixing the pressure contact portion capable of moving close to and away from at an arbitrary position. The container is inserted into the opening of the guide base portion from the bottom side, and the outside of the container Hula The upper surface of the di-section is brought into contact with the step surface of the step portion, a container attachment, characterized in that for mounting the container be nipped from the side in the container the pair of pressure contact portion.

  According to a second aspect of the present invention, in the attachment for a container according to the first aspect, the pressure contact portion supporting means is provided on the side of the guide base portion that protrudes downward from the lower surface of the pressure contact portion, and is long in the approaching / separating direction. The container attachment includes: an extended through hole, wherein the protrusion slides in the through hole when the lower surface of the pressure contact portion slides on the guide base portion.

  According to a third aspect of the present invention, in the container attachment according to the second aspect, the projection is provided with a large-diameter portion, and the guide base portion extends inwardly at a lip of the through-hole of the through hole of the guide base portion The container attachment is characterized in that an inner flange portion is provided, and the inner flange portion covers and is prevented from coming off on a large diameter portion of the projection portion.

  According to a fourth aspect of the invention, in the container attachment according to any one of the first to third aspects, the press contact portion fixing means is configured to elastically urge the press contact portion in the proximity direction. It is the attachment for containers.

  According to a fifth aspect of the present invention, in the attachment for a container according to the fourth aspect, the press contact portion fixing means includes a plunger whose tip is in contact with the press contact portion, and a plunger holding that holds the plunger so as to reciprocate in the approaching and separating directions. A container attachment comprising: a portion and a spring that biases the plunger toward the opposite pressure contact portion.

  According to a sixth aspect of the present invention, in the container attachment according to any one of the first to fifth aspects, the pressure contact surface of the pressure contact portion has a width along the opening, and the pair of pressure contact portion support means spaced apart from each other. It is the attachment for containers characterized by being supported.

  A seventh aspect of the present invention is the container attachment according to any one of the first to sixth aspects, wherein only one of the press contact portions is movable.

  According to an eighth aspect of the present invention, in the container attachment according to any one of the first to seventh aspects, the guide base portion further includes a penetrating liquid supply port and a lid for closing the liquid supply port from above. This is an attachment for containers.

  According to a ninth aspect of the present invention, in the container attachment according to the eighth aspect, the liquid supply port is positioned so that the lens portion of the objective lens used for observation is within the observable range of the opening when viewed from above. Sometimes, the container attachment is designed to be correlated with the opening so as to overlap a rotation locus drawn by rotation of a revolver of the lens unit.

  According to a tenth aspect of the present invention, in the container attachment according to the eighth or ninth aspect, only one of the press contact portions is movable, and the liquid supply port is provided outside the press contact portion on the fixed side. This is an attachment for containers.

  The invention of claim 11 is the container attachment according to any one of claims 1 to 10, wherein the bottom surface of the guide base portion is dimensioned so as to be substantially flush with the bottom surface of the container to be used. It is the attachment for containers characterized by the above.

  A twelfth aspect of the present invention is the container attachment according to any one of the first to eleventh aspects, wherein a space is provided on the guide base portion so as to surround the press-contact portion and a water tank portion is provided, and a water tank that detachably fixes the water tank portion. It is an attachment for containers provided with a part fixing means.

  The invention according to claim 13 is based on temperature information from a temperature detection means in which a transparent surface top heater is provided in a portion corresponding to the observation range in the lid of the storage unit, and the detection unit is inserted into the container. The container attachment according to any one of claims 1 to 12 is detachably provided in a temperature control device of a type that controls the temperature of the top heater to maintain the space in the storage unit at a predetermined temperature or temperature range. A culture apparatus for microscopic observation characterized by the above.

  The invention according to claim 14 is the culture apparatus for microscope observation according to claim 13, further comprising a container holding mechanism for arbitrarily attaching and detaching the container attachment and the container. is there.

  According to the culture apparatus for microscopic observation of the present invention, it is possible to efficiently perform microscopic observation at a high magnification.

It is a disassembled perspective view of the culture apparatus for microscope observation which concerns on embodiment of this invention. It is the perspective view seen from the upper direction of the culture apparatus for microscope observation of FIG. It is the perspective view seen from the downward direction of the culture apparatus for microscope observation of FIG. It is a disassembled perspective view of the attachment for containers and the slide glass plate of the culture apparatus for microscope observation of FIG. It is the perspective view which looked at the state which attached the slide glass plate to the attachment for containers of FIG. 4 from upper direction. It is the perspective view which looked at the state which attached the slide glass plate to the attachment for containers of FIG. 4 from the downward direction. It is a top view which shows the state which mounted | wore the slide glass plate with the attachment for containers of FIG. It is AA sectional drawing of FIG. It is BB sectional drawing of FIG. It is GG sectional drawing of FIG. It is a perspective view which shows the state which mounted | wore the water tank part with the attachment for containers of FIG. It is explanatory drawing of the dimensional relationship of the attachment for containers of FIG. 4, and a slide glass plate. It is explanatory drawing of the dimensional relationship of the attachment for containers of the modification of the attachment for containers of FIG. 4, and a dish. It is a perspective view which shows the state which isolate | separated the cover body from the attachment for containers different from FIG. It is a perspective view which shows the state which closed the attachment for containers of FIG. 14 with the cover body from upper direction. It is a top view which shows the state which mounted | wore the slide glass plate with the attachment for containers of FIG. It is AA sectional drawing of FIG. It is BB sectional drawing of FIG. It is explanatory drawing of the usage method of the attachment for containers of FIG. It is a figure explaining the correlation with the opening part of the attachment for containers of FIG. 14, and a liquid supply port. It is a disassembled perspective view of the conventional container attachment.

A microscopic observation culture apparatus 1 according to an embodiment of the present invention will be described with reference to FIGS.
The microscope observing culture apparatus 1 includes a housing unit 3, a lid 37, a container attachment 45, and the like.
First, the structure of the storage unit 3 will be described.
In FIG. 1, reference numeral 5 denotes a main body portion formed in a substantially rectangular frame shape, and an upper end side opening 7 and a lower surface side opening 9 are formed in the main body portion 5, respectively. As shown in FIG. 1, an inner flange 11 is formed on the lower surface side opening 9 side, and the lower surface side opening 9 is smaller than the upper end side opening 7.

  The side surface of the adapter 13 is fitted and fixed to the edge of the lower surface side opening 9 of the main body 5. The adapter 13 is also formed in a substantially rectangular frame shape, and an inner flange 17 is formed on the lower end side opening 15 side. Reference numeral 19 denotes a presser spring, and the base end of the presser spring 19 is connected to each of four locations on the upper end surface of the adapter 13 so as to be rotatable within the adapter 13. The adapter 13 is a container holding mechanism for mounting a container attachment 45 or a well plate (not shown), and either one can be mounted. According to this mechanism, the container attachment 45 and the like are detachably fitted, and when fitted, they are stably fixed by the elastic force of the presser spring 19 described above.

A concave space defined between the inner surface of the main body 5, the outer surface of the adapter 13, and the upper surface of the inner flange 11 of the main body 5 serves as a water tank 21.
A water supply hose 23 and a gas supply hose 25 are attached to the main body 5, and the tips of the water supply hose 25 enter the water tank 21, and the tip of the gas supply hose 25 extends to the vicinity of its bottom surface.

  Between these, a detection unit holder 31 is provided as an in-container introduction unit for the thin-line thermocouple detection unit 29 of the temperature detection unit 27. The detection unit holder 31 is sandwiched and fixed between the presser screws 33 and 33, and the detection unit 29 is introduced into the main body unit 5, so that the introduction length into the main body unit 5 can be adjusted. It is like that. The detection unit 29 can be bent and is immersed in a culture solution placed in a culture vessel during detection. The main body 5 is also provided with a holder 35 for attaching a pair of perfusion hoses so that perfusion culture is possible.

A lid 37 is provided so as to close the upper end opening 7 of the housing unit 3 having the above structure.
The lid 37 has a substantially rectangular shape, and the frame 39 holds a transparent transparent portion 41 having a rectangular shape. Two transparent glass plates are disposed in the light transmitting portion 41 with a gap therebetween, and a transparent conductive film is formed on the upper surface of the lower transparent glass plate. The transparent conductive film is caused to generate heat by being energized through the electric cord 43, and has a function as a transparent planar top heater.

  Both the electric cord 43 on the lid 37 side and the temperature detection means 27 are connected to a controller (not shown), and the controller uses a feedback method based on temperature information from the temperature detection means 27 and a target temperature. The amount of current supplied to the transparent conductive film is adjusted.

In the accommodation unit 3 having the above-described configuration, the slide glass plate S as a culture container is accommodated using the container attachment 45. The configuration of the container attachment 45 will be described in detail with reference to FIGS.
For convenience of explanation, the vertical direction is defined with the open side of the slide glass plate S as the upper side in FIG.
As shown in FIG. 4, the guide base portion 47 of the container attachment 45 has a rectangular thin plate shape, and a rectangular opening portion 49 is formed in the center portion so as to be substantially similar to the outer rectangular edge. As shown in FIG. 8, a step 51 is formed along the entire circumference of the lip of the opening 49, which is one step higher than the remaining bottom surface. The stepped portion 51 is formed with a shallow concave portion 53 only on one short side.

On the upper surface of the opening 49, mounting recesses 55 and 59 are formed on the opposing long edges. Two screw holes 57, 57 are formed at a distance from each other on the mounting recess 55 side.
Two through holes 61 are formed at a distance from each other on the mounting recess 59 side. The through hole 61 is long in a direction perpendicular to the long side of the guide base portion 47. As shown in FIG. 9, an inner flange 63 is formed on the upper end side of the through hole 61, and the opening is narrowed by the inner flange 63 from the remaining portion. The inner flange 63 extends inwardly from the through hole 61, and the extending edge of the inner flange 63 is similar to the hole edge of the through hole 61.
Fixing holes 65 and 65 are formed at two corners on the long side.
At the four corners of the guide base portion 47, holes are respectively opened, and magnets 67 are fitted and bonded thereto through a silicone adhesive.

A pair of pressure contact portions are provided on the guide base portion 47.
Reference numeral 69 denotes a receiving side pressure contact portion, which has a plate bar shape. Two through holes 71, 71 are formed at a distance from each other in the receiving side pressure contact portion 69. The receiving-side pressure contact portion 69 is positioned by being fitted into the mounting recess 55, and as shown in FIG. 4, the male screw 73 is passed through the through hole 71 with the head facing upward, and the mounting recess By inserting and screwing into the 55 screw holes 57, the receiving side pressure contact portion 69 is fixed in a state where the lower surface thereof is in contact with the upper surface of the guide base portion 47. In the fixed state, a horizontally long plate surface (pressure contact surface) 75 is on the edge of the opening 49. A silicone sheet 77 is affixed to the pressure contact surface 75 via a silicone adhesive.

Reference numeral 79 denotes a presser-side press-contact portion, and in this press-side press-contact portion 79, a thin plate-like rectangular presser portion 83 is integrally formed on one long side of the rectangular main body portion 81. Both end sides of the presser portion 83 extend uniformly from both lateral sides of the main body portion 81. The presser portion 83 is provided with a step, and the lower side protrudes. The protruding side is a plate surface (pressure contact surface) 85. A silicone sheet 77 is also affixed to the pressure contact surface via a silicone adhesive. A knob 87 is provided on the main body 81.
The presser-side press-contact portion 79 is positioned by being brought into and fitted into the opening 49 side of the mounting recess 59, and its lower surface is in contact with the upper surface of the mounting recess 59. The pressure contact surface 85 is on the edge of the opening 49.
The press-contact surface 85 of the presser-side press-contact portion 79 and the press-contact surface 75 of the receiving-side press-contact portion 69 are set to have substantially the same size, and are arranged on the guide base portion 47 so as to sandwich the opening 49 therebetween.

As shown in FIG. 9, two screw holes 89, 89 are formed at a distance from each other on the lower surface side of the presser-side pressure contact portion 79. In the arrangement state described above, the screw holes 89 are recessed for mounting. 59 is in communication with the through hole 61.
As shown in FIG. 6, the male screw 91 as the protruding portion is passed through the long through hole 61 of the mounting recess 59 with the large-diameter head on the lower side, and into the screw hole 89 of the presser-side pressure contact portion 79. Inserted and screwed. At this time, the head 93 of the male screw 91 enters the through hole 61, and the step surface 93 </ b> A of the step formed in the head 93 is slightly below the lower surface of the inner flange 63 of the through hole 61.

The gap between the inner flanges 63 is larger than the stepped portion of the head portion 93 of the male screw 91, and as shown in FIG. Can move in the direction shown. The direction indicated by this arrow is the approaching / separating direction to the receiving side pressure contact portion 69.
The presser-side pressure contact portion 79 moves integrally with the male screw 91. When the presser-side pressure contact portion 79 is gripped by the finger 87 and pulled in the proximity and separation direction, the male screw 91 moves within the movable range. While the lower surface slides on the upper surface of the guide base portion 47, the lower surface moves closer to and away from the receiving side pressure contact portion 69.

The step surface 93A of the head portion 93 of the male screw 91 is always covered with the direction side perpendicular to the approaching and separating direction of the inner flange 63 from above, and is prevented from coming off.
The press contact portion supporting means is configured as described above, and the presser side press contact portion 79 is supported so as to be able to reciprocate in the approaching and separating direction with respect to the receiving side press contact portion 69. The press contact surface 85 has a width in the lateral direction along the opening 49, but is supported by a pair of press contact portion support means spaced apart from each other, and the presser side press contact portion 79 can smoothly move. .
The male screw 91 is accommodated in the through hole 61 and is not an intrusive protrusion.

A fixing portion 95 is provided on the presser side pressure contact portion 79 side. The fixing portion 95 has a “U” shape, and a large surrounding portion 97 is provided at the center. The fixing portion 95 is disposed so as to surround the rectangular main body portion 81 of the presser side pressure contact portion 79 from the lateral side.
As shown in FIG. 10, holes are formed on both sides of the surrounding portion 97 so that a cylindrical plunger holding portion 99 is fitted therein. Each plunger holding part 99 opens to the presser part 83 side and extends long in the approaching / separating direction.

Each plunger holding part 99 houses a coil spring (compression spring) 101, and one end on the bottom side of the plunger holding part 99 is a free end. Reference numeral 103 denotes a plunger, and the base end of the plunger 103 is connected to the other end of the coil spring 101 described above. The distal end of each plunger 103 is bonded to the back surface of the extended portion of the pressing portion 83 with a silicone adhesive.
In the movable range of the presser side pressure contact portion 79, the coil spring 101 is always urged in a direction in which the presser side pressure contact portion 79 is caused to approach the side pressure contact portion 69. The base end side of the plunger 103 is always accommodated and guided in the plunger holding portion 99, and the straight running posture is held.

As shown in FIG. 10, the fixing portion 95 is provided with two screw holes 105, 105 spaced apart from each other, and the fixing portion 95 is moved closer to the outer edge side of the mounting recess 59 and positioned. Each screw hole 105 communicates with the fixing hole 65 of the guide base portion 47.
The male screw 107 is passed through the fixing hole 65 with the head facing down, and is inserted into and screwed into the screw hole 105 of the fixing portion 95 so that the lower surface of the fixing portion 95 is brought into contact with the upper surface of the guide base portion 47. Fixed in contact.
The male screw 107 is accommodated in the fixing hole 65 and is not an obstructive projection.

The container attachment 45 has the above-described configuration, and a method for mounting the slide glass plate S will be described below.
The slide glass plate S is a common one that is commercially available, and has a base portion a that is rectangular and has a shape that is substantially similar to the opening 49 of the guide base portion 47. Four side parts b rise from the inner side of the base part a, although slightly from the edge of the upper surface, and a step part c is formed on the edge side of the base part a. Further, a flap d is continuously provided on one short side. On the upper surface of the flap d, a character for product identification is formed integrally.

First, the knob portion 87 is sandwiched between fingers and pushed in the separating direction, and the presser side pressure contact portion 79 is moved against the spring force of the coil spring 101 to fully open the opening portion 49 of the guide base portion 47. In this state, when the guide base portion 47 is lowered with respect to the slide glass plate S from above and the slide glass plate S is inserted into the opening 49, the upper surface of the stepped portion c of the slide glass plate S becomes the guide base portion. The bottom surface of the slide glass plate S and the bottom surface of the guide base portion 47 are substantially flush with each other in contact with the lower surface of the 47 stepped portion 51. At this stage, there is a slight gap between the side portion b of the slide glass plate S and the side surface of the stepped portion 51, so that it is in a loosely fitted state. Note that the concave portion 53 is opposed to the flap d, and the guide base portion 47 is not lifted even if the character is lifted as described above.
Next, when the force applied to the knob 87 is weakened, the presser-side pressure contact portion 79 moves in the proximity direction, which is the urging direction, so that the slide glass plate S and the pressure-contact surface 85 of the presser portion 83 and the receiving side It is press-clamped between the press-contact surfaces 75 of the press-contact part 69. Thereby, the slide glass plate S is attached to the container attachment 45.
Note that a silicone sheet 77 is attached to each of the pressure contact surfaces 75 and 85 for buffering, and since it is this silicone sheet 77 that directly touches the slide glass plate S, it is difficult to be damaged.

  When the slide glass plate S is to be removed, the knob 87 is sandwiched between fingers and pushed in the separating direction, and the presser side pressure contact portion 79 is moved against the spring force of the coil spring 101 to open the opening 49 of the guide base portion 47. Fully open. In this state, the guide base portion 47 is lifted upward and separated from the slide glass plate S.

FIGS. 2 and 3 show a state in which the slide glass plate S is mounted on the container attachment 45 and then accommodated in the accommodation unit 3 and closed with the lid 37.
Usually, the container attachment 45 is fitted into the storage unit 3, and the slide glass plate S is attached to the container attachment 45.
After closing the lid 37 to define the culture space, carbon dioxide (CO ₂ ) is supplied from the gas supply hose 25, and water is supplied from the water supply hose 23 to the water tank 21. In addition, the culture solution contained in the slide glass plate S is heated from above by energizing the transparent conductive film of the translucent part 41 through the electric cord 43 to generate heat. In addition, although illustration is abbreviate | omitted, the heater is installed also in the bottom part side of the water tank 21, The water in the water tank 21 is evaporated by heating of this heater, and the inside of culture | cultivation space is made into desired humidity. The gas supply hose 25, the water supply hose 23, and the heater described above are also under the control of the controller, and the culture space is maintained constant.

In this microscope observing culture apparatus 1, since the culture solution is heated from above through air, the distance from the heat source is the same regardless of the well w of the slide glass plate S, and the center of the storage unit 3. Whether it is located at the corner or the corner, it is heated evenly. Therefore, the temperature of the culture solution put in all the wells w can be accurately detected by immersing and detecting the detection unit 29 of the temperature detection means 27 in the culture solution put in the wells w in the corners. It can be used for microscopic observation.
The edge of the opening 49 of the container attachment 45 approaches the well w at the corner of the slide glass plate S. The bottom surface of the slide glass plate S and the bottom surface of the guide base portion 47 of the container attachment 45 are as described above. Thus, the objective lens can be sufficiently brought close to the well w at the corner.
Therefore, if the culture apparatus 1 for microscope observation is used, the efficient observation of the microscope observation by high magnification is realizable.

A water tank portion 109 can be attached to the container attachment 45. As shown in FIG. 11, the water tank portion 109 has a “U” shape and is mounted on the guide base portion 47 so as to surround the receiving side press contact portion 69, the presser side press contact portion 79, and the fixing portion 95. . Recesses are respectively formed on the lower surfaces of the four corners of the water tank portion 109, and magnetic pieces (not shown) are inserted therein and bonded via a silicone adhesive. This magnetic piece is flush with the lower surface of the water tank portion 109.
As shown in FIG. 11, when the water tank portion 109 is placed on the guide base portion 47, the magnetic pieces are opposed to the magnets 67 fitted into the four corners of the guide base portion 47. Is adsorbed and the water tank part 109 is fixed on the guide base part 47.

In order to make the slide glass plate S easily enter the guide base portion 47, the slide glass plate S is loosely fitted with a slight gap as described above. If the slide glass plate S satisfies the following relationship with respect to the guide base portion 47 shown in FIG. Therefore, it is possible to cope with a certain degree of difference with respect to standard differences and dimensional errors of commercially available products.

The container attachment 45 is assumed to be a rectangular culture container, but if the shape of the opening portion 49 of the guide base portion 47 is changed to a circular opening portion 113 and the pressure contact surface is formed in an arc shape. A round dish can also be used as an attachment.
Even in this case, as long as the dish D satisfies the following relationship with respect to the guide base portion 47 shown in FIG. Therefore, it is possible to cope with a certain degree of difference with respect to standard differences and dimensional errors of commercially available products.

Next, another example of the container attachment 111 will be described with reference to FIGS.
In addition, the main part is common with the attachment 45 for containers, and description is abbreviate | omitted by attaching | subjecting the same code | symbol to a common part.
As shown in FIGS. 14 and 15, the container attachment 111 is also provided with a receiving-side press contact portion 69 and a presser-side press contact portion 79 with the opening 49 interposed therebetween. And the liquid supply port 113 is formed in the outer side so that the receiving side press-contact part 69 may be enclosed. The liquid supply port 113 penetrates the guide base portion 47 in the vertical direction. A step portion 115 is formed at the mouth edge of the liquid supply port 113, and the upper surface side of the guide base portion 47 is lowered by one step from the upper surface.
Reference numeral 117 denotes a lid, and a knob 119 is attached to the upper surface of the lid 117. The lid body 117 is fitted into the above-described step portion 115 and closes the liquid supply port 113.
The range of the liquid supply port 113 differs depending on the direction as shown in FIGS.

When using a high-magnification objective lens (about 60 to 100 times) during microscopic observation, oil immersion observation or water immersion observation is often performed.
When performing oil immersion observation or water immersion observation, first bring a low-magnification (about 10 to 29 times) objective lens to the observation position, and place a microscope so that the object to be observed is on the optical axis of the objective lens. The observation culture apparatus 1 is positioned. After that, the revolver is rotated to bring the high-magnification objective lens where the low-magnification objective lens is located, but before bringing the high-magnification objective lens to the observation position, A few drops of special oil or water is supplied to the lens unit using a special dropper, and when the objective lens has moved to the observation position, the supplied oil or water is used to move the slide glass plate S. The space between the bottom surface and the lens portion is filled.
In this case, when the container attachment 45 is used, the container attachment 45 is detached from the apparatus main body, and the opening 15 is opened to supply oil or the like. However, the container attachment 45 is removed and reattached. Therefore, there is a possibility that even if a sample such as a cell placed in the well w of the slide glass plate S is small in the well w, the sample may move. Even a slight misalignment becomes a significant problem in high-magnification observation.

In contrast, the container attachment 111 is provided with a liquid supply port 113, through which oil or the like can be dropped and supplied onto the lens portion L of the high-power objective lens, as shown in FIG. When the revolver R is rotated, the objective lens is held up to the observation position while retaining the oil supplied on the lens portion L. If the supply is released for a short time and then closed with the lid 117, the culture space is maintained in a predetermined atmosphere, so that the culture atmosphere is not broken.
In order to make this possible, specifically, as shown in FIG. 20, the entire surface opening portion excluding the step portion 115 of the liquid supply port 113 is the lens portion of the objective lens used for observation as viewed from above. When L is positioned so as to be within the observable range (center, upper left, lower left, upper right, lower right) of the opening 49, a circular rotation locus (dotted line) drawn by the rotation of the revolver R of the lens portion L Is designed to correlate with the opening 49.

Therefore, even if the observation position is changed within the observable range, the trajectory of the lens portion L of the objective lens always crosses the liquid supply port 113. It can be brought to the observation position. In the figure, the thick line portion is a portion that crosses the circular rotation locus.
In this embodiment, since one of the press contact portions is fixed and a space remains on the receiving base press contact portion 69 side on the guide base portion 47, the liquid supply port 113 is designed using that portion. .

As described above, the embodiments of the present invention have been described in detail. However, the specific configuration is not limited to these embodiments, and the invention can be changed even if there is a design change without departing from the gist of the present invention. include.
For example, as long as the culture vessel has a function of allowing the detection part of the temperature detection means to pass through the lid or the container side, a culture vessel closed with a lid can also be used.
In addition, the culture vessel can be adapted to various types by changing the shape of the opening 49 of the guide base portion 47. In addition to the above, for example, two or more dishes are mounted in parallel. You can also.
The container attachment 45 does not always have to be used, and a large one such as a well plate is directly attached to the accommodation unit 3 of the microscope observation apparatus 1.

  The culture apparatus for microscopic observation of the present invention can be used as an accessory of a microscope.

DESCRIPTION OF SYMBOLS 1 ... Culture apparatus for microscope observation 3 ... Housing unit 5 ... Main-body part 7 ... Upper end side opening 9 ... Lower surface side opening 11 ... Inner flange (of lower surface side opening) 13 ... Adapter 15 ... Lower end side opening 17 (of adapter) ... Inner flange (lower end opening) 19 ... Presser spring 21 ... Water tank 23 ... Water supply hose 25 ... Gas supply hose 27 ... Temperature detection means 29 ... (Temperature detection means) Thermocouple type detection part 31 ... Detection part Holder 33 ... Holding screw 35 ... Holder for attaching the hose for perfusion 37 ... Lid body 39 ... Frame body (of lid body) 41 ... Translucent part 43 (of lid body) 43 ... Electric cord 45 ... Attachment for container 47 ... Guide base Portion 49 ... Opening portion (of guide base portion) 51 ... Step portion 53 (of opening portion) ... Recessed portion (of step portion) 55 ... Recessing portion for mounting 57 ... Screw hole 59 ... Recessing portion for mounting 61 ... Through hole 63 ... (Through hole) Inside of the hole Lung 65 ... fixing hole 67 ... magnet 69 ... receiving side pressure contact portion 71 ... through hole 73 ... male screw 75 ... pressure contact surface 77 ... silicone sheet 79 ... presser side pressure contact portion 81 ... main body portion 83 ... presser portion 85 ... pressure contact surface 87 ... Knob 89 ... Screw hole 91 ... Male screw 93 ... (Male screw) head portion 93A, B ... (Head) step surface 95 ... Fixing portion 97 ... (of fixing portion) Enclosing portion 99 ... Plunger holding portion 101 ... Coil spring 103 ... Plunger 105 ... Screw hole 107 ... Male screw 109 ... Water tank part 111 ... Container attachment 113 ... Liquid supply port 115 ... Step part 117 ... Lid 119 ... Knob part S ... Slide glass plate a ... Base part b ... Side Part c ... Step part d ... Flap w ... Well

Claims (14)

A container attachment used to store a container in a storage unit of a culture apparatus for microscopic observation,
A guide base portion having a flat bottom surface, which is fitted into the lower opening portion of the housing unit, an opening portion formed in the guide base portion, and an extra bottom surface provided along the mouth edge of the opening portion. A stepped portion that is one step higher, a pair of pressure contact portions facing each other with the opening interposed therebetween on the guide base portion, and the other pressure contact portion facing at least one of the pressure contact portions in the proximity and separation direction. A pressure contact portion supporting means for reciprocally supporting the pressure contact portion, and a pressure contact portion fixing means for fixing the pressure contact portion capable of moving toward and away from each other at an arbitrary position;
The container is inserted into the opening of the guide base portion from the bottom surface side, the upper surface of the outer flange portion of the container is brought into contact with the step surface of the step portion, and the container is laterally moved by the pair of pressure contact portions. An attachment for a container, wherein the container is attached by being pressed and clamped from above. In the container attachment according to claim 1,
The press contact portion support means includes a protrusion protruding downward from the lower surface of the press contact portion and a through hole provided on the guide base portion side and extending long in the approaching / separating direction, and the lower surface of the press contact portion is the guide base portion The container attachment, wherein the protrusion slides in the through hole when sliding on the top. In the container attachment according to claim 2,
The protruding portion is provided with a large-diameter portion, and an inward flange portion extending inward is provided at a rim of the guide base portion on the side of the approaching and separating direction, the large-diameter portion of the protruding portion. An attachment for a container, wherein the inner flange portion covers and is prevented from coming off. The container attachment according to any one of claims 1 to 3,
The container attachment characterized in that the pressure contact portion fixing means is configured to elastically urge the pressure contact portion in the proximity direction. In the attachment for containers according to claim 4,
The pressure contact portion fixing means includes a plunger whose tip is in contact with the pressure contact portion, a plunger holding portion that holds the plunger so as to be capable of reciprocating in a proximity and separation direction, and a spring that biases the plunger toward the opposite pressure contact portion. An attachment for a container, characterized by comprising: In the container attachment according to any one of claims 1 to 5,
An attachment for a container, wherein the pressure contact surface of the pressure contact portion has a width along the opening and is supported by a pair of pressure contact portion support means spaced apart from each other. The container attachment according to any one of claims 1 to 6,
An attachment for a container, wherein only one of the press contact parts is movable. In the container attachment according to any one of claims 1 to 7,
A container attachment comprising a guide base portion provided with a liquid supply port further penetrating and a lid for closing the liquid supply port from above. The container attachment according to claim 8,
When viewed from above, the liquid supply port overlaps the rotation locus drawn by the rotation of the revolver of the lens unit when the lens unit of the objective lens used for observation is positioned within the observable range of the opening. The container attachment is designed to correlate with the opening. The container attachment according to claim 8 or 9,
Only one of the press contact parts is movable, and the liquid supply port is provided outside the press contact part on the fixed side. In the container attachment according to any one of claims 1 to 10,
A container attachment, characterized in that the bottom surface of the guide base portion is dimensioned so as to be substantially flush with the bottom surface of the container to be used. The container attachment according to any one of claims 1 to 11,
A container attachment comprising: a space for providing a water tank portion on the guide base portion so as to surround the pressure contact portion; and a water tank portion fixing means for removably fixing the water tank portion.   A transparent planar top heater is provided in the portion of the lid of the storage unit corresponding to the observation range, and the top heater is temperature-controlled based on temperature information from temperature detection means that has inserted the detection unit into the container. A microscope characterized in that a container attachment according to any one of claims 1 to 12 is detachably provided in a temperature control device of a type that maintains a space in a storage unit at a predetermined temperature or temperature range. Observation culture device. The culture apparatus for microscopic observation according to claim 13,
A culture apparatus for microscopic observation, comprising a container holding mechanism for arbitrarily attaching and detaching a container attachment and a container.

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