JP3525303B2 - Microbial culture observation container - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microbial culture observation container, which is capable of accurately measuring a microscopic growth process in a state in which a microorganism to be observed is cultured, or observing and measuring a drug effect on the observation target. Used for.

[0002]

2. Description of the Related Art Conventionally, a closed type microorganism culture observation container of this kind has been shown in FIGS. 13 and 14, for example.

The container shown in FIG. 13 is a slide glass 10.
1 and the cover glass 103 are opposed to each other so as to form a predetermined culture space 104, and are fixed with an adhesive 105 or the like to form a closed structure, and an injection needle 107, 1 is provided in the flow direction of the culture solution.
09 is inserted to supply and discharge the culture solution. Reference numeral 111 is a microorganism to be observed, and 113 is a microscope.

The container 115 shown in FIG. 14 is made of a transparent transparent body, and a culture space 104 for placing the microorganism 111 to be observed is provided between the upper surface 117 and the lower surface 119. The supply path 121 and the discharge path 123 are provided.

The culture space 104 of each of the vessels is very narrow in order to facilitate observation. That is, in the case of a microorganism that grows so that its roots grow like molds, if the culture space 104 is widened as shown in FIG. 12b, the focus of the microscope may shift gradually. In such a case, for example, it becomes inconvenient when the automatic setting is performed, and since it is difficult to measure the three-dimensional growth, the numerical value such as the growth rate is also erroneously measured.
In order to prevent this, the culture space 104 was made narrow (FIG. 12a).

[0006]

However, such a narrow culture space has the following drawbacks. That is, air bubbles may enter and flow into the culture solution. In such a case, the air bubbles do not easily flow, and sometimes the air bubbles accumulate and become large, which is an obstacle to observation.

If air bubbles are accumulated, the culture solution may not come into contact with the inoculated microorganism to be observed. In this case, enzymes and nutrients are not supplied, or the drug solution does not come into contact with the microorganism. Sometimes it was impossible to measure.

By the way, the flow of the liquid flowing in the space forms a thin layer called a boundary layer near the surface of the object due to the viscosity of the object, but when the space becomes minimal, the entire space becomes the boundary layer, and as a result, flows there. Due to the high frictional resistance of the liquid, the flow is greatly deteriorated. If air bubbles are present in the culture medium in such a state, the flow of the culture medium is further hindered and it becomes difficult to transfer the liquid.

For these reasons, there has been a certain limit in forming a narrow culture space, and as a result, it is still not possible to observe extremely small microorganisms such as molds and yeasts while flowing a culture solution or a drug. It was insufficient.

The present invention solves the above-mentioned disadvantages, prevents the obstruction of culture observation and measurement due to air bubbles, and improves the flow of the culture solution, thereby making it possible to form a narrow culture space. It is an object of the present invention to provide a microorganism culture observation container capable of observing even minute microorganisms such as mold and yeast while flowing a culture solution or a drug.

[0011]

To achieve the above object, the present invention provides a closed vessel having a culture / observation section having a space for culturing microorganisms and having an observation window made of a transparent body,
In the microorganism culture observation container configured to supply and discharge the culture solution to the culture / observation section, a bypass is provided outside the supply path and the discharge path of the culture solution connected to the culture / observation section, There is provided a microorganism culture observation container having an upper surface set higher than the upper surface of the culture / observation section. In this case, it is conceivable that a bypass is provided in the container body. Further, it is considered that the bypass has a narrower discharge side than the supply side of the culture solution. In addition, it is conceivable to provide a rectification section in a narrowly formed bypass (narrow path) and communicate with the discharge path via the rectification section. At this time, it is conceivable that the rectifying section is provided so as to be inclined at an acute angle with respect to the flow direction of the culture solution. In addition, it is conceivable to provide a rectifying unit in a widely formed bypass (broad path) and communicate with the supply path via the rectifying unit. At this time, it is conceivable that the rectifying section is provided with an obtuse angle with respect to the flow direction of the culture solution. Further, it is possible to attach a road width adjusting member to the narrow road. Further, it is conceivable that the culture / observation section, the supply passage, and the discharge passage are formed by a spacer separate from the container body and the container body. It is also conceivable that the spacer is a transparent plate-shaped body having a predetermined thickness, and the ink is applied to the portion other than the portion corresponding to the culture / observation portion and the portion forming the supply passage and the discharge passage. Also,
It is conceivable to provide commutators for narrowing the path width at the inlet side end and the outlet side end of the culture solution of the printing spacer.
At this time, it is conceivable that a dot-shaped portion was printed on the portion corresponding to the culture / observation portion to such an extent that observation was not hindered. At this time, it is conceivable that the number of dot portions is one or two or more. Further, it is conceivable that the spacer is a molded spacer made of a plate-shaped body having a predetermined thickness in which a portion corresponding to the culture / observation part is cut out. At this time, it is conceivable to provide commutators at the inlet side end and the outlet side end of the culture medium of the shaped spacer, respectively, to narrow the path width. It is also possible that the commutator is a curved one. Further, it is conceivable to provide the container body with a notch corresponding to the culture / observation section, and a supply path and a discharge path. At this time, it is conceivable to provide commutators for narrowing the path width at the inlet side end and the outlet side end of the culture solution of the container body. At this time, it is possible that the commutator is curved. In addition, it is conceivable to make the culture / observation part a very small space.

[0012]

In the microorganism culture observation container according to the present invention, even if bubbles are mixed in the culture solution, these bubbles pass through the bypass before the culture solution flows into the supply passage, and are therefore removed here. That is, since the upper surface of the bypass is set higher than the upper surface of the culture / observation section which is at the same height as the supply passage, the bubbles float above the culture solution and are discharged from the outlet through the bypass. . As a result, there are no obstacles for culturing / observation and measurement due to air bubbles, and the flow of the culture solution is not disturbed by the air bubbles and is good.

[0013]

Embodiments of the microorganism culture observation container according to the present invention will now be described in detail with reference to the embodiments shown in the drawings. For convenience of explanation, the same reference numerals are given to the parts having the same functions.

In FIGS. 1 to 4, reference numeral 1 denotes a container body made of an appropriate material, a central portion of which is cut out, and a culture / observation portion 3 is formed therein. That is, a watertight packing 5, a transparent plate 7 made of glass, a spacer 9 to be described later, and a transparent plate 11 made of glass are sequentially laminated in this order from the top in the central notch, and the transparent plate 7 and the transparent plate 11 are combined. Culture space 12 between
Are formed, and this serves as the culture / observation section 3. Reference numeral 3a is an observation window of the culture / observation section 3, and the transparent plate 7 and the transparent plate 11 are provided.
Formed in a circle on top. Outside the culture / observation section 3,
The culture / observation section 3 is provided with a culture solution supply path 13 and discharge path 15 of the same height, and is connected / communicated to the culture / observation section 3.

Outside the supply passage 13 and the discharge passage 15, there are provided bypasses 17 and 19 whose upper surface is set higher than the upper surface of the culture / observation section 3, and through the discharge passage 15 and the supply passage 13. It communicates with the culture / observation section 3.

The bypass is on the discharge side (narrow path) 17
Is narrower than that on the supply side (broad path) 19, a pressure difference occurs hydrodynamically, so that the flow of the culture solution is further improved, which is desirable.

The bypass (narrow path) 17 is provided with a rectifying section 21 composed of a large number of projections inclined at an acute angle with respect to the flow direction of the culture solution, and the bypass (narrow path) is provided via the rectifying section 21. 17 and the discharge path 15 are communicated with each other. By doing this, the bypass (narrow road) 17 and the bypass (wide road) 19
Since the pressure difference between and becomes higher, the suction of the discharged culture solution is further improved, which is desirable.

Further, a path width adjusting member 23 made of a rod-like member is installed in a portion of the bypass (narrow path) 17 adjacent to the bypass (wide path) 19, and culture / observation can be performed by adjusting the width of this member. The flow rate of the culture solution is adjusted by adjusting the flow rate ratio of the culture solution flowing through the section 3 and the bypasses 19 and 17.

Reference numeral 25 is a hose fitting serving as an inlet of the culture solution, and 27 is a hose fitting serving as an outlet of the culture solution, which are connected to a bypass (broad path) 19 and a bypass (narrow path) 17, respectively.

On the back surface of the container body 1 having the above-mentioned structure, a back lid 31 similarly having a notched central portion is fastened and fixed by a screw 33 via a watertight packing 29 having a notched central portion.

Next, the operation and effect of the microorganism culture observation container having the above structure will be described. First, the transparent plate 11
Inoculate with the microorganism to be observed, or insert it between the transparent plate 7 and the transparent plate 11 and then install in a container,
The culture solution is fed by press fitting or suction. The culture solution fed from the hose fitting 25 is supplied from the supply path 13 to the culture space 12 of the culture / observation section 3, and then discharged from the hose fitting 27 via the discharge path 15. In this case, even if the culture solution contains air bubbles,
These bubbles pass through the bypass (broad path) 19 before the culture solution flows into the supply path 13, and are removed here.
That is, since the upper surface of the bypass (broad path) 19 is higher than the upper surface of the supply path 13, the bubbles float above the upper surface of the bypass (broad path) 19 from the culture solution, and the bypass (broad path) 19 bypasses the upper surface. It is discharged from the hose fitting 27 via the narrow path 17.

As a result, the bubbles do not flow into the culture space 12 of the culture / observation section 3 from the supply passage 13, so that the flow of the culture solution is good and the observation obstacles due to the bubbles are prevented. Further, since the flow of the culture solution becomes good, it becomes possible to form a narrower culture space, and even microbes such as mold and yeast can be observed while flowing the culture solution and the drug. is there. Furthermore, since the contact of the culture medium with the microorganisms to be observed is prevented due to the presence of air bubbles, nutrients and enzymes are smoothly supplied to the microorganisms to be observed, and the microscopic growth process in the cultured state can be prevented. It can be measured accurately. Further, in the case of drug efficacy measurement, the drug solution contained in the culture solution is brought into sufficient contact with the microorganism to be observed, so that it is possible to prevent the measurement failure due to poor contact.

Since all the above-mentioned parts can be disassembled, disinfection and cleaning can be easily performed.

The microorganism culture observation container according to the present invention is not limited to the above embodiment. For example, the bypass need not have a difference in width.

Further, even when the bypass has a difference in width, it is possible to adjust the flow rate ratio between the culture space and the bypass only by adjusting the size of the narrow path with respect to the wide path without providing the width adjustment member 23. Is.

Further, the culture / observation section, the supply passage and the discharge passage may be formed by a spacer separate from the container body and the container body, or the container body may be integrally provided with the spacer function.

5 to 7 show examples of spacers. FIG. 5 shows a printing spacer 9a in which ink is applied by printing to form a frame 35 on a transparent plate 11 made of glass having a predetermined thickness. That is, since ink is present in a portion other than the portion corresponding to the culture / observation portion 3 on the transparent plate 11 and the portion where the supply passage 13 and the discharge passage 15 are formed, the ink is raised by the thickness of the ink from other portions. The culture / observation section 3, the supply path 13 and the discharge path 15 are thereby formed.

The printing spacer 9a is also provided with a commutator 37 formed by printing a large number of bar codes. This is to prevent the watertightness from being lowered due to the pressure of the packing 5 and the packing 29 when the thin glass is used as a base, and to impart a rectifying function. The thinner the glass, the smaller the distance between barcodes.

The printing spacer 9a is further printed with a dot-like portion 39 consisting of small dots that do not interfere with the observation at the portion corresponding to the culture / observation portion 3. This is because, when a thin glass is used as a base, when the culture solution is pressed into the culture space 12, the upper and lower transparent plates 7 and 11 bulge in a protruding arc shape and bubbles 41.
Are likely to remain (FIG. 11b), and conversely, when the culture solution is sucked and discharged from the culture space 12, the upper and lower transparent plates 7 and 11 are dented in a concave arc shape to further narrow the space (FIG. 11c). When a pump with a pulsating flow such as a peristal or a plunger is used during such press-fitting and inhaling, the glass plate surface vibrates, making it difficult to observe while the culture solution is flowing. In order to prevent such a defect, the dot-like portions 39 are appropriately arranged. In this case, the transparent plate 7,
The deformation of 11 is small (Fig. 11a), and the flow of the culture solution is good. Further, it is possible to cope with pulsating flow, and the appearance of the culture / observation section 3 is improved.

The above-mentioned parts 35 of the printing spacer 9a,
Instead of printing, 37 and 39 may be provided by film sticking. In this way, if printing or film sticking is used, it is possible to deal with the case where the culture space 12 is a micro space of a micron level and mass production is possible.

FIG. 6 shows a molded spacer 9b made of a plate-like body 43 with a predetermined pressure in which a portion corresponding to the culture / observation section 3 is cut out.
Is. The molding spacer 9b is used when the height of the culture space 12 and the flow paths 13 and 15 is increased. The molded spacer 9b is also provided with a commutator 45 composed of a large number of projecting pieces for narrowing the road width as in the above-mentioned embodiment, and when the plate-like body 43 is thin, the watertightness caused by bending due to crimping is provided. It is possible to further improve the flow of the culture solution by preventing the decrease of the amount of water and imparting a rectifying action.

FIG. 7 shows a modified example in which the commutator is bent. When the commutator 37a is configured in this way, when the transparent plate 11 serving as the base is thin, the contact area between the packing and the transparent plate is large, so that the glass plate is less likely to be deformed, which contributes to improvement of watertightness. Is. The configuration in which the commutator is bent in this way can also be applied to the commutator 45 of the molded spacer 9b in FIG.

Next, in FIGS. 8 to 10, the notch 47 corresponding to the culture / observation section 3 and the supply path 13 are provided in the container body 1a.
It is an embodiment in which the discharge passage 15 and the discharge passage 15 are provided, which is, so to speak, a case where the spacer function is integrally formed in the container body. This embodiment also shows a case where the bypass 20 is not provided with a wide difference and a case where neither a rectifying unit nor a commutator is provided. 49 is an air reservoir and 51 is a front cover.

Even if the container body is integrally formed with the spacer function as in this embodiment, a rectifying section or a rectifier may be provided. In this case, the rectifier is as shown in FIG. It is also possible to bend it.

The microorganism culture observation container according to the present invention is particularly effective when the culture space 12 is an extremely small space at the micron level. That is, when air bubbles enter the culture space 12, the air bubbles often do not flow with a small amount of liquid transfer (described above). Especially, when the spacer is thinned and the culture space 12 is formed in the above-mentioned micron-level extremely small space, the culture is performed once. The bubbles mixed in the space 12 will not move. Therefore, conventionally, it was not possible to transfer the solution to the culture space formed in the microscopic micro space. However, according to the microorganism culture observation container according to the present invention, even if the culture space 12 is formed in such an extremely small space, the culture space 1
If air bubbles are not mixed in 2 and the bypass is widened or narrowed, the flow of the culture solution is promoted, so that the above obstacles are eliminated. In addition, in the above-described embodiment, the minimum space of the culture space 12 is, for example, about 30 μm.

Furthermore, if the bypass is set higher than the supply passage and the discharge passage, the supply passage and the discharge passage do not necessarily have to be set at the same height as the culture / observation section.

The rectifying section 21 may be provided not only on the discharge path 15 side but also on the supply path 13 side.

For the constituent members such as the container body and the spacer, an appropriate material such as resin or metal can be selected according to the purpose.

[0039]

As described above, according to the microorganism culture observation container of the present invention, bubbles do not enter the culture / observation section, so that the obstacles for culture / observation due to bubbles or the measurement defects caused by bubbles can be prevented. can do. In addition, since it is possible to prevent liquid feeding inhibition due to air bubbles in a narrow culture space, the flow of the culture liquid becomes good, and as a result, it becomes possible to form a narrower culture space, and it is possible to minimize mold and yeast. Even such microorganisms can be observed while flowing a culture solution or a drug.

[Brief description of drawings]

FIG. 1 is a plan view showing an embodiment of a microorganism culture observation container according to the present invention.

2 is a cross-sectional view of the EFGH of FIG.

FIG. 3 is an ABCD cross-sectional view of FIG.

FIG. 4 is an enlarged view of part X in FIG.

FIG. 5 is a perspective view showing an example of a spacer.

FIG. 6 is a perspective view showing another example of a spacer.

FIG. 7 is a perspective view showing a modified example of the spacer when the rectifying unit is bent.

FIG. 8 is a plan view showing another embodiment of the microorganism culture observation container according to the present invention.

9 is a JK sectional view of FIG.

10 is a sectional view taken along line LM in FIG.

FIG. 11 is a diagram used for explaining the operation of one embodiment of the microorganism culture observation container according to the present invention.

FIG. 12 is a diagram used for explaining a conventional example.

FIG. 13 is a diagram used for explaining a conventional example.

FIG. 14 is a diagram used for explaining a conventional example.

[Explanation of sign]

1 container body 1a Container body 3 Culture and observation department 3a Observation window 5 packing 7 transparent plate 9 spacers 9a printing spacer 9b Molded spacer 11 transparent plate 12 culture space 13 supply path 15 discharge path 17 Bypass (narrow) 19 Bypass (wide road) 20 Bypass 21 Rectifier 23 Road width adjustment member 25 Hose fitting 27 Hose fitting 29 packing 31 case back 33 screws 35 frames 37 Commutator 37a commutator 39 dots 41 air bubbles 43 Plate 45 commutator 47 notch 49 Air reservoir 51 front cover 101 slide glass 103 cover glass 104 culture space 105 adhesive 107 injection needle 109 injection needle 111 microorganisms 113 microscope 115 containers 117 upper surface 119 lower surface 121 Supply path 123 discharge path

─────────────────────────────────────────────────── ─── Continuation of the front page (56) References Tokuhyo 7-506256 (JP, A) (58) Fields investigated (Int.Cl. ⁷ , DB name) C12M 1/00-3/10 C12M 1 / 00-7/08 PubMed BIOSIS / WPI (DIALOG)

Claims (21)

(57) [Claims] 1. A closed vessel is provided with a culture / observation section having a space for culturing microorganisms and having an observation window made of a transparent body so that a culture solution is supplied to and discharged from the culture / observation section. In the constructed microbial culture observation container,
A microorganism culture observation container characterized in that a bypass is provided outside a supply path and a discharge path of a culture solution connected to an observation section, and an upper surface of the bypass is set higher than an upper surface of the culture / observation section. 2. The microorganism culture observation container according to claim 1, wherein a bypass is provided in the container body. 3. The microorganism culture observation container according to claim 1 or 2, wherein the bypass has a narrower discharge side than the supply side of the culture solution. 4. The microorganism culture observation container according to claim 2 or 3, wherein a narrowed bypass (narrow passage) is provided with a rectifying portion, and the rectification portion communicates with the discharge passage. . 5. The microorganism culture observation container according to claim 4, wherein the rectifying section is provided with an inclination at an acute angle with respect to the flow direction of the culture solution. 6. The microorganism culture observation container according to any one of claims 2 to 5, wherein a rectifying section is provided in a widely formed bypass (broad path), and a microorganism communicating with the supply channel through the rectifying section. Culture observation container. 7. The microorganism culture observation container according to claim 6, wherein the rectifying section is provided with an obtuse angle with respect to the flow direction of the culture solution. 8. The microorganism culture observation container according to claim 4 or 5, wherein a path width adjusting member is attached to the narrow path. 9. The microorganism culture observation container according to any one of claims 1 to 8, wherein the culture / observation section, the supply passage and the discharge passage are formed by a spacer separate from the container body and the container body. Microorganism culture observation container. 10. The microorganism culture observation container according to claim 9, wherein the spacer is made of a transparent plate having a predetermined thickness, and is a portion other than the portion corresponding to the culture / observation portion and the portion forming the supply passage and the discharge passage. Microorganism culture observation container that is a printing spacer with ink applied to the surface. 11. The microorganism culture observation container according to claim 10, wherein commutators for narrowing the path width are respectively provided at the inlet side end and the outlet side end of the culture solution of the printing spacer. 12. The microorganism culture observation container according to claim 10 or 11, wherein a dot-like portion that does not interfere with observation is printed on a portion corresponding to the culture / observation portion. 13. The microorganism culture observation container according to claim 12, wherein the microorganism culture observation container has one dot. 14. The microorganism culture observation container according to claim 12, wherein the microorganism culture observation container has two or more point portions. 15. The microorganism culture observation container according to claim 9, wherein the spacer is a molded spacer made of a plate-shaped body having a predetermined thickness with a portion corresponding to the culture / observation portion cut out. 16. The microorganism culture observation container according to claim 15, wherein a commutator for narrowing the path width is provided at each of an inlet side end and an outlet side end of the culture solution of the molding spacer. 17. The microorganism culture observation container according to any one of claims 10 to 16, wherein the commutator is a bent one. 18. The microorganism culture observation container according to any one of claims 1 to 8, wherein the container body is provided with a notch corresponding to a culture / observation portion and a supply passage and a discharge passage. 19. The microorganism culture observation container according to claim 18, wherein commutators for narrowing the width of the passage are provided at the inlet side end and the outlet side end of the culture solution of the container body. 20. The microorganism culture observation container according to claim 18 or 19, wherein the commutator is a bent one. 21. The microorganism culture observation container according to any one of claims 1 to 20, wherein the culture / observation part has an extremely small space.