JP2004309136A - Container and method of interaction - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cylindrical container for accommodating a liquid and immersing a solid in the liquid to cause an interaction between the liquid and the solid, and to reconcile stable rotation based on positioning accuracy with ease of container attachment/detachment while utilizing the container and its stable rotation to clean the solid, to cause the solid to react with the liquid, and to measure liquid components. <P>SOLUTION: A mesh part removably meshing with a mechanism for rotating the container from below is provided in at least a part of a hollow part constituting an inner surface of a projection part. Via the mesh part, the container is rotated together with the liquid. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a container for washing a solid surface, accelerating a reaction, and performing a measurement using the reaction.
[0002]
[Prior art]
In causing an interaction between a solid surface and a liquid in contact with the solid surface, it is widely practiced to promote the interaction between the solid surface and the liquid by giving a flow to the liquid. The interaction and the process involving the interaction include washing, adsorption, and chemical reaction. In addition, various methods can be considered as a method of giving a flow to the liquid, but in a wide-mouthed container, the contained liquid is stirred by some method to create a liquid flow, and the target solid is immersed here. That is common.
[0003]
As one of the stirring methods, the container is frequently rotated. That is, in this stirring method, a fluid is generated in the liquid near the inner wall of the container by the viscosity of the liquid in the container by applying a rotational driving force of a certain number of rotations from the outside of the container through a rotating mechanism coaxial with the center axis of the container. Then, the whole liquid is stirred based on this flow. Patent Document 1 discloses a method for dispersing fine particles in a liquid by this method.
[0004]
In addition, although different from so-called stirring, the target solid is allowed to stand still in contact with the liquid, such as by suspending it in the liquid in the container, and by rotating the container, the liquid is brought into contact with the solid. Can be relatively fluid. In the method described in Patent Document 2, a sensor is efficiently brought into contact with a sample solution by this method, and the sensor is used for measurement such as fluorescence immunoassay.
Since these methods do not require a stirring blade or a stirrer, they are advantageous in maintenance of the apparatus, prevention of contamination of the sample, and the like, and also have an advantage that the target liquid can be easily exchanged.
[0005]
[Patent Document 1]
JP 2002-196006 A [Patent Document 2]
JP 2002-257732 A
[Problems to be solved by the invention]
However, in the methods of Patent Literatures 1 and 2, a cup-shaped receiver is used for placing the container and transmitting the rotational driving force. If the receiver is designed to be large, the placement of the container in the receiver becomes unstable, which may hinder the rotation of the container or cause the container to fall. Such instability of mounting and rotation of the container is caused by anisotropic centrifugal force generated when the center axis of the container is slightly eccentric with respect to the rotation axis of the receiver, A mechanism is required that can simply and reliably align the center axis of the container with the rotation axis of the receiver.
[0007]
The present invention has been made in view of the above circumstances, and in particular, for a solid suspended and immersed in a liquid in a cylindrical container from above, the container is rotated to make the liquid relative to the solid. It is an object of the present invention to promote the interaction between the solid (surface) and the liquid by giving a proper flow, and to perform an effective interaction.
[0008]
[Means for Solving the Invention]
That is, the present invention is a container for containing a liquid, immersing a solid in the liquid, and allowing the components in the liquid to interact with the solid, the container comprising:
(1) a cylindrical outer peripheral wall having a bottom at the lower end and an open upper end;
(2) a projection extending from the bottom in the opening direction and having an outer shape as a rotating body concentric with a cylindrical central axis of the outer peripheral wall;
(3) a liquid storage portion formed as a space between the outer peripheral wall and the projection;
(4) a cavity having an opening on the outer surface of the bottom portion and extending into the projection;
(5) a meshing portion provided on at least a part of the hollow portion, a rotating shaft for rotating the container is detachably fitted, and the meshing portion can mesh with the projection. For containers.
[0009]
In the present invention, it is preferable that at least a part of the cavity has a tapered shape in the central axis direction.
In the present invention, it is preferable that the engagement portion has a rib.
In the present invention, it is preferable that the height of at least a part of the rib is continuously reduced downward in the direction of the central axis of the projection.
In the present invention, it is preferable that the number of the ribs is 2 to 4.
In the present invention, it is preferable that the container further contains a polypropylene resin composition.
[0010]
The present invention is a method of containing a liquid, immersing a solid in the liquid, and performing an interaction between components in the liquid and the solid,
The container is
(1) a cylindrical outer peripheral wall having a bottom at the lower end and an open upper end;
(2) a projection extending from the bottom in the opening direction and having an outer shape as a rotating body concentric with a cylindrical central axis of the outer peripheral wall;
(3) a liquid storage portion formed as a space between the outer peripheral wall and the projection;
(4) a cavity having an opening on the outer surface of the bottom portion and extending into the projection;
(5) a meshing portion provided on at least a part of the hollow portion, a rotating shaft for rotating the container is detachably fitted, and can mesh with the projection;
And a rotating shaft for rotating the container is joined to a meshing portion of the container, and the interaction is performed by rotating the container through the meshing portion. .
Preferably, the invention further comprises that the interaction comprises washing the solid surface with the liquid.
Preferably, in the present invention, the interaction includes a reaction between the solid surface and a component in the liquid.
[0011]
In the present invention, it is preferable that the amount of the component in the liquid is measured by the reaction when the interaction is performed.
In the present invention, it is further preferable that the reaction at the time of performing the interaction is a reaction of capturing an antigen in the liquid with at least one type of antibody previously immobilized on the surface of the solid.
In the present invention, it is preferable that the antigen used in the reaction is at least one of a bacterium and a virus.
[0012]
BEST MODE FOR CARRYING OUT THE INVENTION
The container of the present invention has a cylindrical outer shape, the solid suspended from above in the liquid in the container, for a solid immersed, to rotate the container to give the liquid a relative flow to the solid, The purpose is to promote the interaction between the solid (surface) and the liquid. Here, the interaction means a reaction between a component in a liquid and a solid surface, adsorption of a liquid component to the solid surface, washing of the solid surface, elution or extraction from the solid surface into the liquid, and the like.
[0013]
FIG. 1 is a perspective view showing an example of the container of the present invention, and FIG. 2 is a sectional view (cut along a section including the central axis of the container) and a bottom view of an example of the container of the present invention. The container 1 has a liquid container 21 provided inside a cylindrical outer peripheral wall, and a projection 2 is provided at the center of the liquid container 21. The protrusion 2 has an outer shape of a rotating body about the cylindrical central axis of the container 1, and extends in the opening direction from the bottom surface of the liquid container 21 along the central axis. Due to the presence of the projections 2, when the liquid container 21 is viewed from the upper surface of the container, the liquid container 21 has a ring shape, and when the liquid in the container is given a flow, the liquid flows through the ring-shaped flow path. The solid suspended and immersed from above can efficiently contact the liquid flowing with the rotation of the container in the narrowed flow channel. The projection 2 also functions as filling an extra space and minimizing the volume of the liquid container 21 when the amount of the liquid to be stored is set to a necessary minimum. Further, the projection 2 has a top surface 3 which is a measure of the liquid level when a predetermined volume of liquid is put into the container by adjusting the top surface 3 to the liquid level.
[0014]
The interior of the projection 2 forms a cavity 5, and the bottom surface of the cavity 5 is an opening 4. An engagement portion is formed in the hollow portion 5. The meshing portion may be formed in the entire cavity 5 or may constitute a part of the cavity 5. The main feature of the present invention is that a mechanism for rotating the container from below into the cavity 5 through the meshing portion in the cavity can be engaged. This meshing method makes it easy to attach and detach the rotation mechanism to and from the container, and at the same time improves the positioning accuracy of the center axis of the container.
[0015]
It is preferable that at least a part of the cavity 5 is provided with a tapered shape 23 such that the inner diameter of the projection 2 is reduced from below to above in the central axis direction of the container. The tapered portion preferably accounts for 50 to 100% by volume of the entire projection. The angle of the tapered wall surface is preferably 1 to 30 degrees. By providing this tapered shape 23, even if the difference between the inner diameter of the projection 2 and the outer diameter of the rotating mechanism inserted into the cavity 5 from below is reduced at the contact point, it will not be possible to attach or detach the container. Less. Further, the center axis of the container 1 can be accurately aligned with the center axis of the rotation mechanism, and stable rotation of the container can be obtained. The rotation may be performed continuously in only one direction, or the rotation direction may be intermittently reversed at predetermined intervals.
Since there is no shielding on the outer surface of the container, the container can be easily gripped regardless of whether it is manual or automatic.
[0016]
The hollow portion 5 is provided with a meshing portion for transmitting the rotational force from the rotation mechanism. A preferred form of the meshing portion is a rib 6 provided in the hollow portion 5. This one embodiment is shown in FIG. A pyramid-shaped part is prepared on the rotating mechanism as the container mounting part 7, and this part fits into the hollow part 5 of the container 1. At the start of rotation of the rotation mechanism, the side wall of the pyramid-shaped portion is fixed by the rotational driving force of the container while being in contact with the rib 6 of the container 1, and the rotational driving force is transmitted from the rotating mechanism to the container through this contact portion. . Other forms of the meshing portion include a prismatic shape and a ratchet mechanism, but a rib structure is preferable because it is easy and effective to manufacture.
Regarding the shape of the rib, at least the height of a part of the rib (the length in the direction perpendicular to the axial direction of the projection) is continuously reduced toward the lower part in the axial direction of the projection (the overhang is reduced). (Smaller) (25 in FIG. 3).
[0017]
This is because, when the container 1 is placed, when the pyramid-shaped rotating mechanism side portion as shown in FIG. 3 comes into contact with the end surface of the rib 6, the contact surfaces slide obliquely with each other. This is because it is possible to transit to an optimally engaged state of the rotation mechanism and the rib. The number of ribs may be one or more, but is preferably two to four in view of ease of container attachment and detachment and transmission of rotational driving force.
[0018]
The size of the container depends on the size and shape of the solid to be immersed, and is not particularly limited, but the width and depth of the flow path surrounded by the outer peripheral wall and the protrusion are different from those of the solid. There must be no collision and a gap that gives an effective relative flow velocity to the solid surface. In addition, even if the outer peripheral wall has the same inner diameter, by increasing the outer diameter of the projection, the amount of liquid that can be accommodated decreases, and the relative flow velocity of the liquid and the solid during rotation of the container increases. Therefore, it is necessary to set the outer diameter of the outer peripheral wall and the inner diameter of the projection in accordance with the amount of liquid to be handled, and to set the number of revolutions so as to obtain a desired relative flow velocity.
[0019]
In order to use a solid having a large surface area and to minimize the amount of liquid to be used, (outer diameter of projection) / (inner diameter of outer peripheral wall) is preferably 0.2 to 0.8, and more preferably 0.4 to 0.8. 6 is more preferred. (Protrusion height) / (Accommodation depth) is 0.5 to 1 in order to make the liquid level a guide, to stably engage the rotating mechanism and the meshing portion, and to perform stable rotational driving. 0.0 is preferable, and 0.7 to 0.9 is more preferable. Further, in order to obtain a desired liquid flow rate and to minimize the amount of liquid to be used, (depth of the housing portion) / (inner diameter of the outer peripheral wall) is preferably 0.5 to 3.0, and more preferably 1.0 to 3.0. -2.5 is more preferable.
[0020]
The material of the container should be selected according to the liquid used and the components present in the liquid.However, if the system involves proteins such as an antigen-antibody reaction, and it is desirable to be disposable for testing or the like, less adsorption is required. It is desirable that the material is a material containing an inexpensive polypropylene resin composition.
Further, the container of the present invention can be molded by a known method, but preferably, an injection molding method is used.
[0021]
The interaction in the present invention is performed by rotating the container in the manner described above, thereby giving a relative flow of the liquid to a solid suspended and immersed in the liquid in the container from above. Suitable modes of interaction include (1) washing the surface of the solid with the liquid, (2) reacting the surface of the solid with components in the liquid, and (3) from the solid surface to the liquid. Elution or extraction. According to the interaction method of the present invention, by flowing the liquid, the contact between the liquid and the solid is effectively performed, so that such an interaction can be effectively performed. These interactions may be performed alone or in combination.
[0022]
The term “reaction” used herein includes non-chemical reactions such as physical adsorption and extraction, and chemical reactions involving covalent bonds, coordination bonds, and electron transfer. Further, the solid surface used as a measurement probe can be washed with a liquid having a detergent such as a surfactant.
In addition, for example, adsorption of an organic compound contained in an aqueous solution on a solid surface made of activated carbon, a chain forming reaction between a chelating agent supported on the solid surface and metal ions in a liquid, formation of a dissolved portion of the solid surface from a resist film, Examples include elution into a liquid, and capture of an antigen in a liquid by an antibody carried on a solid surface.
[0023]
Using the reaction as a means for measuring the amount of a specific component in a liquid is useful as one embodiment of the present invention. That is, the above-described container structure and the stable rotation mechanism based on the meshing method are suitable for easily ensuring sensitivity, quantification, and reproducibility in measurement for performing various quantitative analyses. Further, the fact that the container can be easily attached and detached is highly convenient when a large number of samples are exchanged and handled. It is desirable to use a solid immersed in a liquid sample as a measurement probe, and to measure components in the liquid concentrated on the surface thereof by an appropriate means on the spot or after moving to another location. If necessary, a suitable capturing function such as an adsorbent or a receptor may be provided on the surface of the measurement probe.
[0024]
The components in the liquid, that is, the measurement target include solutes such as molecules and ions, and dispersed fine particles, and are particularly effective for fine particles having a small self-diffusion. Examples of such fine particles include heavy metal fine particles, bacteria, and viruses. In other words, for small molecules with high mobility due to thermal perturbation in liquids, access to the measurement probe surface is somewhat easy without the flow of liquids. In addition, the flow of the liquid facilitates the approach to the measurement probe, so that quantitative analysis excellent in sensitivity, quantitativeness, and reproducibility can be performed. Therefore, a preferred embodiment of the present invention includes an immunoassay in which an antigen in a liquid sample is captured by an antibody previously immobilized on the surface of a measurement probe, and in particular, the antigen is at least one of bacteria and viruses having low self-diffusion. One of the measurements is: Bacteria include Escherichia coli, Salmonella, Staphylococcus aureus and the like. Viruses include hepatitis virus, influenza virus and the like. It is preferable to use antibodies corresponding to the antigens or a part thereof, respectively. These bacteria and viruses may be measured alone, or two or more bacteria and viruses may be measured by combining these antibodies.
[0025]
【Example】
(Example)
By injection molding of a polypropylene resin composition, the inner volume of the shape shown in FIG. ) And a protrusion height of 35 mm). An example in which a fluorescent immunoassay measurement was performed using this will be described. This example includes, as examples of the interaction method of the present invention, washing of a measurement probe with a washing solution in a container, and capture reaction of an antigen in a sample solution.
As a measurement probe, a polystyrene resin probe 9 having a shape shown in FIG. 4 (length: 44 mm, fiber-shaped portion length: 41 mm, fiber-shaped portion diameter: 0.7 mm) was used. The probe comprises a flange portion 12 for suspending the probe when immersed, a lens-like portion 11 for condensing fluorescence, and a fiber-like portion 10. An anti-Escherichia coli O157: H7 polyclonal antibody is immobilized on the surface of the fiber-shaped portion 10 of the probe. The optical system shown in FIG. 5 was used for excitation and photometry. The light (635 nm) of the semiconductor laser 18 passes through the half mirror 17 and enters from above the probe 9. The return light from the probe is extracted in the horizontal direction by the half mirror 17 and introduced into the spectrophotometer 20 via the optical fiber 19.
[0026]
The measurement port 13 is filled with the cleaning liquid, and excitation and photometry are performed in a state where the probe is immersed in the liquid. As a washing solution, a 0.01 M phosphate buffer (pH 7.4) containing 0.5% by mass of polyoxyethylene (20) sorbitan monolaurate was used. The fluorescent-labeled antibody was prepared by allowing Cy5 bisfunctional reactive dye (manufactured by Amersham Biosciences) to act on the antibody, and was used by dissolving the fluorescent-labeled antibody in the washing solution at 2 μg / ml. As a specimen, a specimen containing Escherichia coli O157: H7 at 100 CFU / ml, 1000 CFU / ml and 10,000 CFU / ml in water was used. The containers A and B use the containers of the present invention, respectively.
[0027]
First, the following operation was performed on the probe as a blank measurement.
(1) Immersion in a labeled antibody solution for 5 minutes.
(2) The container A is immersed in the container A containing only the cleaning liquid, and the container A is continuously rotated in the same direction at 100 rpm for 10 minutes.
(3) Move to the measurement port for excitation and photometry.
[0028]
Next, the following operation was performed on the probe as a main measurement.
(1) The container B is immersed in the container B containing the sample, and the container B is continuously rotated in the same direction at 100 rpm for 10 minutes.
(2) Immersion in the labeled antibody solution for 5 minutes.
(3) The container A is immersed in the container A containing only the cleaning liquid, and the container A is continuously rotated in the same direction at 100 rpm for 10 minutes.
(4) Move to the measurement port for excitation and photometry.
[0029]
This measurement was continuously performed four times. The blank measurement signal was subtracted from the main measurement signal to obtain a net fluorescence signal.
For a sample containing E. coli O157: H7 at 100 CFU / ml, 1000 CFU / ml, and 10,000 CFU / ml, a signal increase was obtained in proportion to the number of times of measurement, and each signal change rate was determined by the number of bacteria (CFU / ml). Was proportional.
[0030]
(Comparative example)
The same washing liquid, sample, and measurement probe as in Example 1 were used, and the container used was a cylindrical one having no projection. Further, the measurement was carried out by the same fluorescence immunoassay as in Example 1 except that the container was not stirred during the washing of the measurement probe and the reaction for supplementing the antigen.
[0031]
This measurement was continuously performed four times. For samples containing 1000 CFU / ml and 10,000 CFU / ml Escherichia coli O157: H7, respectively, a signal increase was obtained in proportion to the number of times of measurement, but for samples containing 100 CFU / ml Escherichia coli O157: H7. No quantitative signal increase was obtained in proportion to the number of measurements.
[0032]
【The invention's effect】
In order to immerse the solid in the liquid contained in the container and allow the liquid to interact with the solid, the container is rotated to give the liquid a relative flow to the solid. The rotation and the ease of attaching and detaching the container can both be ensured. Furthermore, from the above effects, when measuring the amount of components in the liquid, the sensitivity, quantitativeness, reproducibility, and convenience of measurement can be improved.
[Brief description of the drawings]
FIG. 1 (a) is a perspective view from above of an example of the container of the present invention. FIG. 1B is a perspective view from below of an example of the container of the present invention.
FIG. 2A is a cross-sectional view of the container of the present invention cut along a plane including a central axis. FIG. 2B is a schematic diagram illustrating the bottom surface of the container of the present invention.
FIG. 3 is a schematic view showing an engagement state between a container and a rotation mechanism according to the present invention.
FIG. 4 is a schematic diagram showing a measurement probe in an example.
FIG. 5 is a schematic diagram showing an optical system in an example.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Container 2 Projection part 3 Projection top surface 4 Opening 5 Cavity part 6 Rib 7 Container mounting part 8 Pyramid part 9 Optical probe 10 Fiber part 11 Lens part 12 Flange part 13 Measurement port 14, 15, 16 Lens 17 Half mirror 18 Semiconductor laser 19 Optical fiber 20 Spectrophotometer 21 Liquid container 22 Part where height decreases in the central axis direction of rib 23 Tapered shape

Claims (12)

A container for containing a liquid, immersing a solid in the liquid, and allowing an interaction between components in the liquid and the solid,
The container is
(1) a cylindrical outer peripheral wall having a bottom at the lower end and an open upper end;
(2) a projection extending from the bottom in the opening direction and having an outer shape as a rotating body concentric with a cylindrical central axis of the outer peripheral wall;
(3) a liquid storage portion formed as a space between the outer peripheral wall and the projection;
(4) a cavity having an opening on the outer surface of the bottom portion and extending into the projection;
(5) a meshing portion provided on at least a part of the hollow portion, a rotating shaft for rotating the container is detachably fitted, and can mesh with the projection;
A container comprising: The container according to claim 1, wherein at least a part of the hollow portion has a tapered shape in the central axis direction. The container according to claim 1, wherein the engagement portion has a rib. 4. The container according to claim 3, wherein the height of at least a part of the rib is continuously reduced downward in a central axis direction of the protrusion. 5. The container according to claim 3 or 4, wherein the number of the ribs is 2 to 4. The container according to any one of claims 1 to 5, wherein the container includes a polypropylene resin composition. A method for containing a liquid, immersing a solid in the liquid, and performing an interaction between components in the liquid and the solid,
The container is
(1) a cylindrical outer peripheral wall having a bottom at the lower end and an open upper end;
(2) a projection extending from the bottom in the opening direction and having an outer shape as a rotating body concentric with a cylindrical central axis of the outer peripheral wall;
(3) a liquid storage portion formed as a space between the outer peripheral wall and the projection;
(4) a cavity having an opening on the outer surface of the bottom portion and extending into the projection;
(5) a meshing portion provided on at least a part of the hollow portion, a rotating shaft for rotating the container is detachably fitted, and can mesh with the projection;
An interaction method, characterized in that a rotating shaft for rotating the container is joined to a meshing portion of the container, and interaction is performed by rotating the container via the meshing portion. The method of claim 7, wherein the interaction comprises cleaning the solid surface with the liquid. 9. The interaction method according to claim 7, wherein the interaction includes a reaction between the solid surface and a component in the liquid. The interaction method according to claim 9, wherein the amount of the component in the liquid is measured by the reaction. The interaction method according to claim 9 or 10, wherein the reaction is a reaction of capturing an antigen in the liquid with at least one type of antibody previously immobilized on the solid surface. The interaction method according to claim 11, wherein the antigen is at least one of a bacterium and a virus.

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