WO2015152219A1 - 電極、複合電極、液体分析計 - Google Patents
電極、複合電極、液体分析計 Download PDFInfo
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- WO2015152219A1 WO2015152219A1 PCT/JP2015/060080 JP2015060080W WO2015152219A1 WO 2015152219 A1 WO2015152219 A1 WO 2015152219A1 JP 2015060080 W JP2015060080 W JP 2015060080W WO 2015152219 A1 WO2015152219 A1 WO 2015152219A1
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- insertion hole
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/333—Ion-selective electrodes or membranes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/283—Means for supporting or introducing electrochemical probes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/301—Reference electrodes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/302—Electrodes, e.g. test electrodes; Half-cells pH sensitive, e.g. quinhydron, antimony or hydrogen electrodes
Definitions
- the present invention relates to an electrode, a composite electrode, and a liquid analyzer used for pH measurement, ORP measurement, ion concentration measurement such as sodium and potassium, and the like.
- a pH electrode has been used to measure the pH of a sample solution used in, for example, experiments.
- This pH electrode is inserted into a container in which a sample solution is placed and immersed in the sample solution to measure pH and the like. May have adverse effects.
- the tip of the pH electrode protrudes into the container, it is necessary to clean it carefully so as not to damage the pH electrode.
- the pH electrode is liable to be adhered to the pH electrode, making the cleaning operation more complicated.
- Patent Document 1 describes a pH electrode provided in a culture medium container. This pH electrode is inserted from the side of the container to measure the pH of the culture solution, but since the tip of the pH electrode protrudes into the container, the pH electrode is used for cleaning the inside of the container. There is a problem that the pH electrode needs to be washed carefully so as not to be damaged, and that the pH electrode is liable to be adhered, and the pH electrode is complicated to clean. Further, when the culture solution is agitated, the cell hits the tip, causing a problem that stress is applied to the cell.
- the present invention provides an electrode that prevents the contents of the sample solution from adhering to the electrode and preventing the inside of the container from being easily washed when the pH is measured by immersing the electrode in the sample solution in the container. Is the main issue.
- the present invention provides an electrode that can be used very suitably for measurement of pH and the like of cell culture fluids.
- the electrode according to the present invention is an electrode that is inserted into an insertion hole provided in a container in which a sample liquid to be measured is placed, and includes an inner electrode, a container that accommodates the inner electrode, and the inner electrode.
- a gel-like or liquid internal liquid filled in the container so as to be in contact with the internal electrode, and the container is electrically connected to the internal electrode via the internal liquid and detects the potential of the sample liquid
- the container can be positioned by the positioning surface so that the distal end surface of the container and the inner wall surface of the container are flush with each other.
- the electrodes can be cleaned, and the inside of the container can be easily cleaned.
- this electrode can be used very suitably for pH measurement of a cell culture solution. That is, since the front end surface of the container and the inner wall surface of the container are in the same plane, the electrode may protrude into the container, or the step formed in the container may be exposed to provide the insertion hole. Absent. Therefore, when the cell culture solution placed in the container is stirred, the cell does not hit a part of the protruding electrode or the stepped portion, and the cell is not subjected to stress.
- the tip surface itself is formed in a flat shape, even when the amount of the sample solution is small, when the sensor unit is brought into contact with the sample solution, the sample solution spreads over the entire sensor unit. As a result, the pH of the sample solution can be measured.
- the container has an outward surface facing the direction in which the container is inserted into the insertion hole, and the positioning surface is A configuration in which the container is positioned at the mounting position by abutting on the outward surface is mentioned.
- the electrode according to the present invention is characterized in that the container includes a protruding portion protruding in a direction orthogonal to the insertion direction, and the positioning surface is formed on the protruding portion. .
- the positioning body is formed in the protrusion part which protrudes in the direction orthogonal to an insertion direction, the positioning surface is a container when an electrode is inserted in the insertion hole of a container.
- the electrode is brought into contact with the outward surface of the electrode and insertion of the electrode stops there.
- the front end surface of a container and the inner wall face of a container can be made into the same planar shape.
- the electrode according to the present invention is configured such that the container is fixed to the insertion hole when an annular member fitted around the protruding portion presses the protruding portion against the container. It is characterized by.
- the annular member can be provided separately from the container, the container can be formed smaller than when the container is provided with the annular member. Therefore, the electrode itself can be formed small, for example, even when the insertion hole of the container is small, this electrode can be suitably attached.
- the electrode according to the present invention is characterized in that the container is inserted in close contact with the insertion hole.
- the container is inserted into the insertion hole without any gaps, so that a clean state can be maintained.
- a sample solution such as a cell culture solution does not enter between the container and the insertion hole, or cells do not get caught in the portion and accumulate, it is possible to prevent adverse effects on the cell culture. it can.
- the internal electrode and the internal liquid are an internal electrode and an internal liquid for a comparative electrode for measuring a reference potential of the sample liquid, and the sensor unit is a liquid junction part.
- the internal electrode and the internal liquid may be an internal electrode and an internal liquid for an ion-selective electrode that measures the potential of the sample solution, and the sensor unit may be a response membrane.
- the composite electrode according to the present invention includes a support having a response membrane, a second internal liquid filled in the support, and a second internal electrode immersed in the second internal liquid.
- a composite electrode comprising an ion-selective electrode configured, and an electrode whose internal electrode and internal liquid are for a comparison electrode, wherein the response membrane is provided on the front end surface of the support, The response film and the liquid junction are provided on the same plane.
- the electrode is a comparative electrode, the electrode is an ion-selective electrode, or the composite electrode is provided with a reference electrode and an ion-selective electrode. it can. Since such a comparative electrode, an ion selective electrode, or a composite electrode has the above-described electrode configuration, it can be suitably used for pH measurement of a cell culture solution using these.
- the above-described composite electrode a conversion device that converts measurement information from the composite electrode into desired information, and a display device that displays information converted by the conversion device are provided.
- Liquid analyzers If such a liquid analyzer is used, since the front end surface of the composite electrode is flush with the inner wall surface of the container, the above-described effects can be obtained.
- the electrode is attached to the attachment position so that the tip surface of the electrode and the inner wall surface of the container are substantially flush with the container, so that a clean state can be maintained. it can. Also, for example, when the cell culture solution is agitated, there is no stress applied to the stepped part formed in the container in order to provide part of the electrode from which the cell protrudes and the insertion hole, so that the cell culture is adversely affected It is possible to measure pH while performing cell culture without affecting.
- the inner wall surface of the container is substantially flush, the inside of the container can be easily cleaned. Furthermore, since the cells are not caught by the protruding electrode or the exposed stepped portion, the pH can be measured while the culture medium is smoothly stirred and cell culture is performed.
- the tip surface itself is formed in a flat shape, even when the amount of the sample solution is small, when the sensor unit is brought into contact with the sample solution, the sample solution spreads over the entire sensor unit. As a result, the pH of the sample solution can be measured.
- the whole schematic diagram of the electrode in one embodiment of the present invention The whole schematic diagram of the measurement system in the embodiment.
- FIG. 1 shows an electrode 100 according to this embodiment.
- the electrode 100 is described as the composite electrode 6 including the comparison electrode 2 and the ion selective electrode 4, but the electrode 100 is not limited to the composite electrode, and the electrode 100 may be used as the comparison electrode 2 or the ion selective electrode 4, respectively. These electrodes may be used.
- 2 and 3 show a state in which the electrode 100 is inserted into an insertion hole 12 provided in a container 10 in which a sample solution 8 to be measured such as a cell culture solution is placed.
- the comparison electrode 2 includes an inner electrode R for the comparison electrode 2, a container 14 that houses the inner electrode R, and a comparison electrode that fills the container 14 so as to be in contact with the inner electrode R.
- 2 is a sensor that detects the reference potential of the sample liquid 8 and is provided on the tip surface 18 of the container 14 and electrically connected to the internal pole R via the internal liquid 16.
- a liquid junction part 22 as a part.
- the ion selective electrode 4 includes a support body 26 having a response membrane 24, a second internal liquid 28 for the ion selective electrode 4 filled in the support body 26, The second internal electrode M for the ion-selective electrode 4 immersed in the internal liquid 28 is provided.
- a response film 24 is provided on the front end surface 30 of the support 26, and the sample liquid 8 is provided by the response film 24. The electric potential according to the ion concentration of is measured.
- the composite electrode 6 includes the comparison electrode 2 and the ion selective electrode 4 as described above, and the liquid junction portion 22 of the comparison electrode 2 and the response membrane 24 of the ion selective electrode 4 are provided on the same plane. It has been. Specifically, the composite electrode 6 is provided with the container 14 so as to surround the support 26 and integrally includes the ion selective electrode 4 and the comparison electrode 2. Rotating body shape.
- the ion selective electrode 4 has a cylindrical shape, and a flat response glass is provided as a response membrane 24 on the tip surface 30 of a resin support 26.
- the support 26 is filled with a predetermined second internal liquid 28 for ion-selective electrodes, and an internal electrode M composed of a silver / silver chloride electrode is immersed in the second internal liquid 28. Yes.
- the comparison electrode 2 has a resin container 14 formed in a cylindrical shape like the support 26 so as to surround the outer periphery of the support 26, and between the support 26 and the container 14.
- the space provided in is provided so as to form a sealed space.
- the cylindrical container 14 is composed of a container body 15 having an opening formed at the rear end face facing the front end surfaces 18 and 30, and a lid 36 that seals the opening.
- the lid 36 has an outer diameter larger than the outer diameter of the opening of the container body 15, and when the lid 36 is attached to the container body 15, the outer peripheral edge of the lid 36 is the container body.
- the protrusions 38 protrude in the radial direction from the outer peripheral surface of the outer periphery 15.
- the radial direction is a direction orthogonal to the rotation axis of the container 14, and in this embodiment, the direction orthogonal to the direction in which the container 14 is inserted into the insertion hole 12 (hereinafter also referred to as the insertion direction). is there.
- the insertion hole 12 is an internal space of the attached portion X that is formed as a part of the container 10 and to which the composite electrode 600 is attached.
- the attached portion X of the present embodiment has a cylindrical shape protruding outward from the side wall of the container 10, and a space surrounded by the inner peripheral surface X ⁇ b> 1 of the attached portion X is the insertion hole 12. It is formed as.
- the inner peripheral surface X1 of the mounting portion X has a tapered shape that gradually decreases in diameter from the outside to the inside of the container 10.
- the step part is formed in the inner wall surface 32 of the container 10 by the internal peripheral surface X1 of the said to-be-attached part X by the structure mentioned above.
- the container 14 has a positioning surface Y for positioning the container 14 at an attachment position where the tip surfaces 18 and 30 and the inner wall surface 32 of the container 10 are flush. is doing.
- the positioning surface Y is a surface that comes into contact with an outward surface formed in the container 10 that faces away from the insertion direction, and the container 14 is inserted into the insertion hole 12 so that the outer surface faces the outer surface. It is a surface that receives a force opposite to the insertion direction.
- the inner peripheral surface X1 of the mounted portion X and the outer end surface X2 of the mounted portion X described above are formed as the outward surfaces, and the container body 15 that abuts each of these outward surfaces.
- the outer peripheral surface Y1 and the inward surface Y2 of the protrusion 38 are the positioning surface Y. More specifically, the entire outer peripheral surface Y1 is in close contact with the entire inner peripheral surface X1, and the inwardly facing surface Y2 is in contact with the outer end surface X2, whereby the container 14 is positioned at the mounting position. .
- the outer peripheral surface Y1 of the container body 15 has a shape corresponding to the inner peripheral surface X1 of the mounted portion X, and in this embodiment, has a tapered shape that gradually decreases in diameter from the rear end toward the front end. Eggplant.
- the front end surfaces 18 and 30 of the container 14 are flush with the inner wall surface 32 of the container 10, that is, substantially flush with each other.
- the term “substantially flush” as used herein is not necessarily limited to the state in which the tip surfaces 18 and 30 and the inner wall surface 32 are strictly flush with each other, but the contents of the sample solution 8 (for example, a sample solution as a cell culture solution). 8 includes a state in which the distal end surfaces 18 and 30 and the inner wall surface 32 are slightly shifted in the insertion direction to the extent that the cells and the like included in 8 are not caught by the above-described stepped portion or container 14.
- the displacement between the distal end surfaces 18 and 30 of the container 14 and the inner wall surface 32 of the container 10 is within 5 mm by attaching the container 14 to the attachment position.
- the contents of the sample solution 8 are not easily caught on the above-described stepped portion, and the tip surfaces 18 and 30 can be made difficult to get dirty.
- the deviation is more preferably within 2 mm. This is because if the deviation is larger than 2 mm, there arises a problem that cells, microorganisms, etc.
- the composite electrode 6 further includes an annular member 42 that is externally fitted to the protruding portion 38 in order to attach the container 14 to the attachment position described above, and the annular member 42 attaches the protruding portion 38 to the container 10.
- each positioning surface Y comes into contact with each outward surface, and the container 14 is positioned at the mounting position.
- the annular member 42 has a screw portion 46 that is screwed with a screw portion 44 provided at the distal end portion of the outer peripheral surface of the attached portion X, and when these screw portions 44 and 46 are screwed,
- the outer peripheral surface Y1 of the container body 15 is in close contact with the inner peripheral surface X1 of the mounted portion X, and the inwardly facing surface Y2 of the protruding portion 38 is in contact with the outer end surface X2 of the mounted portion X so as to be stored.
- the body 14 is fixed to the insertion hole 12.
- the container body 15 is provided with an opening (not shown) for injecting the internal liquid 16 first on the side wall thereof, and the internal liquid 16 is injected into the container 14 from the opening. After that, the sealed state is maintained by sealing the opening.
- the internal liquid 16 is a gel-like or liquid KCl solution having a predetermined concentration, and a gel-like or liquid one is selected depending on the measurement conditions and the like.
- a flat liquid junction portion 22 is provided on the distal end surface 18 of the container 14, and the liquid junction portion 22 is formed of a porous body such as polyethylene, ceramic, Teflon (registered trademark), or the like. It is. The internal liquid 16 flows out from such a liquid junction 22.
- an internal electrode R composed of a silver / silver chloride electrode is provided so as to be immersed in the internal liquid 16.
- the internal electrode R and the internal electrode M of the ion-selective electrode 4 are connected to an external potentiometer (not shown) via wiring, and the pH of the sample solution is measured by this potentiometer.
- the connection portion is filled with, for example, an adhesive or the like without any gap so that the hermeticity of the sealed space is ensured.
- the container 14 is inserted in close contact with the insertion hole 12.
- the shape of the insertion hole 12 is a columnar shape
- the shape of the container 14 fits in the columnar space without a gap. It is formed to have a columnar shape.
- the shape of the insertion hole 12 is not limited to a columnar shape, and may be a rectangular shape.
- the shape of the container 14 may be formed so as to be in close contact with such a shape.
- This measuring system 200 is inserted into the insertion hole 12 provided in the container 10 so that the container 10 into which the sample liquid 8 to be measured is put, the liquid junction 22 and the response film 24 are immersed in the sample liquid 8. And a composite electrode 6 having a reference electrode 2 and an ion selective electrode 4 attached thereto.
- the composite electrode 6 is connected to an external potentiometer, a recording device, etc., and the pH of the sample solution 8 is measured by this potentiometer, and the pH value is recorded. Recorded on the device.
- the measurement system 200 includes a container 10 having an opening 48 in the upper part, and the container 10 is particularly preferably used for cell culture, and may be a resinous material. It does not have to be a thing that deteriorates and impairs its function. Moreover, if it is a thing made from glass, the material deterioration by radiation sterilization, such as a gamma ray, can be suppressed.
- the container 10 can be used for a sample solution other than the cell culture solution.
- a stirring blade (not shown) for stirring the sample solution 8 is provided in the container 10 so as to extend upward from the bottom thereof.
- a rotating shaft such as a motor (not shown) is further inserted below the stirring blade, and the stirring blade is rotated by the rotation of the rotating shaft to stir the sample liquid 8.
- a lid member 52 is provided at the opening 48 of the container 10 so that the inside of the container 10 can be kept in a sealed state.
- the lid member 52 is provided with tubes 54 and 56 penetrating at the tip thereof.
- a sample solution 8 such as a cell culture solution is introduced from the tube 54 into the container 10, and cell culture is conducted from the tube 56.
- the buffer solution necessary for the flow is introduced.
- the gaps between the tubes 54 and 56 and the lid member 52 are not caused by, for example, an adhesive.
- the cell culture solution is heated to a temperature suitable for culture, for example, about 30 ° C. to 50 ° C. . Then, cell culture is performed while the stirring blade in the container 10 is stirring the cell culture solution, and the pH of the cell culture solution is measured by the composite electrode 6 attached to the side surface of the container 10.
- the liquid analyzer 400 includes a composite electrode 6, a conversion device 72 that converts measurement information from the composite electrode 6 into desired information, and a display device 74 that displays information converted by the conversion device 72. .
- the composite electrode 6 and the conversion device 72 are connected by a signal cable 76 having a predetermined length.
- the signal cable 76 transmits the potential of the sample solution 8 such as a cell culture solution detected by the composite electrode 6 to a conversion device 72 configured by an arithmetic processing device or the like, and the conversion device 72 converts the potential data to desired data such as a pH value. Converted.
- Data such as the pH value is displayed by a display device 74 such as a liquid crystal display, whereby the operator can check the pH value of the sample liquid 8.
- the liquid analyzer 400 When the liquid analyzer 400 is used, it is operated by a plurality of buttons 78 provided on the conversion device 72. Each function is assigned to each button 78, for example, there are functions such as calibration, measurement start, and measurement end. By pressing the measurement start button, the pH measurement of the sample solution 8 is started and the measurement is ended. Pressing the button ends the pH measurement. By setting the composite electrode 6 of the liquid analyzer 400 in the insertion hole 12 of the container 10, it is possible to measure the pH of the cell culture solution without applying stress to the cells while performing cell culture.
- the conversion device 72 and the display device 74 are provided separately from the composite electrode 6.
- the present invention is not limited thereto, and the conversion device 72 is downsized and provided inside the composite electrode 6. Only the display device 74 may be provided outside the composite electrode 6. Further, the display device 74 may also be provided integrally with the composite electrode 6. With such a configuration, the size of the liquid analyzer 400 itself can be reduced, and the configuration of the measurement system can be simplified.
- the composite electrode 6 can be used very suitably for pH measurement of the cell culture solution. That is, since the composite electrode 6 is substantially flush with the inner wall 32 of the composite electrode 6 and the tip surfaces 18 and 30 of the composite electrode 6 with respect to the container 10, the composite electrode 6 protrudes into the container 10, The step portion formed to provide the insertion hole 12 is not exposed. Therefore, when the cell culture solution put in the container 10 is stirred, the cells do not hit the composite electrode 6 and the cells are not stressed. When cells are stressed, the culture is adversely affected. However, since this is not the case, pH measurement can be performed while performing cell culture.
- the comparison electrode 2 and the ion selective electrode 4 are resinous, but can be sterilized so as not to adversely affect the cell culture.
- Other materials may be used as long as they do not exist. For example, when it is made of glass, material deterioration can be suppressed even by sterilization by radiation such as gamma rays, and the function as a composite electrode can be maintained.
- the pH can be measured while performing the cell culture by stirring the culture solution smoothly.
- the front end surfaces 18 and 30 and the inner wall surface 32 of the container 10 become substantially flush, the inside of the container 10 can be easily cleaned.
- the composite electrode 6 can also be used for pH measurement of sample solutions other than the cell culture solution.
- the front end surfaces 18 and 30 of the composite electrode 6 are formed in a flat shape, the liquid junction 22 and the response film which are sensor units even when the amount of the sample liquid 8 is small.
- the sample solution 8 spreads over the entire sensor unit, whereby the pH of a small amount of sample solution can be measured.
- the outer peripheral surface Y1 of the container body 15 is in close contact with the inner peripheral surface X1 of the mounted portion X, and the inward surface Y2 of the protruding portion 38 is mounted. Since it abuts against the outer end surface X2 of the portion X, the insertion of the composite electrode 6 stops there, whereby the front end surfaces 18 and 30 of the container 14 and the inner wall surface 32 of the container 10 become substantially flush.
- the protrusion part 38 should just protrude so that it may contact
- the container 14 of the above embodiment has the outer peripheral surface Y1 of the container body 15 and the inward surface Y2 of the protrusion 38 as the positioning surface Y, but only one of them. May be. Specifically, for example, when the lid portion 36 does not have the protruding portion 38, the container 14 has only the outer peripheral surface Y1 of the container body 15 as the positioning surface Y. On the other hand, when the inner peripheral surface X1 of the attached portion X and the outer peripheral surface Y1 of the container body 15 are not tapered, and the force opposite to the insertion direction does not act on the outer peripheral surface Y1, the container 14 has only the inward-facing surface Y2 of the protrusion 38 as the positioning surface Y.
- the container 14 may further include one or a plurality of positioning surfaces separately from the two positioning surfaces Y in the embodiment.
- the container 14 is fixed to the insertion hole 12 by the screw portion 44 of the attached portion X and the screw portion 46 of the annular member 42 being screwed together.
- the part or the whole may be fixed to the attached part X by, for example, heat welding.
- part or all of the positioning surface Y and the attached portion X are preferably formed from the same material, and specifically, for example, those formed from a resin such as polyethylene are preferable.
- the positioning surface Y is configured to abut against the container 10 so that a force opposite to the insertion direction is applied to the positioning surface Y.
- a frictional force is generated between the surface Y1 and the inner peripheral surface X1 of the mounted portion X, and this frictional force acts on the outer peripheral surface Y1 of the container body 15 as a force opposite to the insertion direction. You may do it.
- the outer peripheral surface Y1 of the container body 15 and the outer surface Y1 of the container body 15 are generated so that the frictional force described above is generated at the attachment position where the distal end surfaces 18 and 30 of the container 14 and the inner wall surface 32 of the container 10 are in the same plane.
- the friction surface should just be formed in the internal peripheral surface X1 of the to-be-attached part X. FIG. In this manner, with the container 14 attached to the attachment position, the container 14 is securely fixed to the attachment position by fitting, for example, a rubber cap member from the outside of the container 14. be able to.
- the cap member is fixed in a state where an adhesive or the like is applied to the back surface of the cap member described above, and the container 14 is pulled toward the cap member together with the signal cable 76, for example. You may make it fix to a cap member. In this case, it is not necessary to form the friction surface described above, and the container 14 can be easily fixed at the mounting position.
- the composite electrode 6 has the annular member 42 as described above, the annular member 42 can be provided separately from the housing body 14, and the housing body 14 itself includes the annular member 42. Compared to the case, the container 14 can be formed smaller. Therefore, the composite electrode 6 itself can be formed small. For example, even when the insertion hole 12 of the container 10 is small, the composite electrode 6 can be suitably attached.
- annular member 42 is provided with a flange portion protruding radially at the peripheral edge portion thereof, and the composite electrode 6 is attached to the attachment position by fixing the flange portion to the container 10 with a screw member or the like. It may be done.
- the container 14 since the container 14 is inserted so as to be in close contact with the insertion hole 12, in such a case, the container 14 is inserted into the insertion hole 12 without a gap. Since the sample solution such as the sample liquid does not enter between the container 14 and the insertion hole 12, or the cells are not caught in the portion and collected, the adverse effect on the cell culture can be prevented.
- the outer shape of the container 14 may be formed in a truncated cone shape, and may be various shapes such as a columnar shape and a rectangular shape, or any shape that can be inserted into the insertion hole 12 closely.
- the container 14 forms a sealed space, once the reference electrode 2 is sterilized, the sterilized state is maintained. For example, when cell culture is performed for several weeks to several months, the pH of the cell culture solution is measured. In the inside, there is no fear of invasion of germs and the like from the outside into the reference electrode 2, and it is possible to prevent the cell culture from being adversely affected.
- the container 14 has a closed closed space formed by the lid portion 36, once the composite electrode 6 is sterilized, the sterilized state is maintained, and the pH of the cell culture solution is measured. There is no risk of invasion of germs from the outside, and it is possible to prevent adverse effects on cell culture.
- the electrode 100 may be the comparison electrode 2 or the ion selective electrode 4.
- the electrode 100 is the comparative electrode 2
- the internal electrode and the internal liquid for the comparative electrode are used, and the sensor part is provided with a liquid junction part formed of a porous body such as polyethylene.
- the electrode 100 is the ion selective electrode 4
- the internal electrode and the internal liquid are used for the ion selective electrode, and a flat glass response film is provided as a response film in the sensor unit.
- the glass responsive film has a small amount of drift compared to an ion sensor such as ISFET even when irradiated with gamma rays, and can be measured with high accuracy, particularly in applications requiring sterilization. I understood.
- a container 62 into which a sample liquid 8 to be measured is placed is formed from a resinous bag-like one.
- the bag-like container 62 is formed so as to have durability so that the culture solution does not leak even if the cell culture solution is placed and the cell culture is performed for several weeks to several months.
- the container 62 can of course be used for a sample solution other than the cell culture solution.
- An insertion hole 64 for inserting the composite electrode 6 is provided on the side surface of the bag-like container 62. When the composite electrode 6 is inserted into the insertion hole 64 and fixed to the insertion hole 64 by the annular member 42.
- this composite electrode 6 is also connected to an external potentiometer, recording device, etc., and the pH of the cell culture solution is measured by this potentiometer, and the pH value is recorded. Recorded on the device
- a tube 60 is provided on the upper surface of the container 62 so that the tip of the tube 60 penetrates and communicates with the inside of the container 62.
- a sample solution 8 such as a cell culture solution or a buffer necessary for cell culture is provided. Liquid or the like flows into the container 62.
- the tube 60 is prevented from generating a gap with the container 62 by an adhesive or the like.
- the cell culture medium is stirred, for example, by shaking the container 62 itself. Also in such a measurement system 300, since the front end surfaces 18 and 30 of the composite electrode 6 are substantially flush with the inner wall surface of the container 62, the inside of the container 62 can be easily cleaned during cleaning. Can be kept clean. In addition, when the cell culture solution is stirred, it is possible to prevent the cells from being subjected to stress by hitting the composite electrode 6 and adversely affecting the cell culture.
- the insertion hole of the composite electrode 6 has the above-described configuration, even such a bag-like container 62 can be provided in any part.
- the location is not limited, and the composite electrode 6 can be installed at a desired location where pH measurement is desired.
- the electrode 100 may be the comparison electrode 2 or the ion selective electrode 4 described above.
- the electrode according to the present invention can be used in various fields. For example, if an insertion hole is provided in a pipe used in various processes and the electrode according to the present invention is inserted into the insertion hole, the pH or the like of drainage or clean water flowing through the pipe can be measured. When culturing a biological tissue such as a microorganism in a medium contained in a petri dish, an insertion hole is provided in the side wall of the petri dish, and the electrode according to the present invention is inserted into the insertion hole, thereby observing the microorganism or the like with a microscope. The pH of the medium can be measured without disturbing.
- an electrode that can be used extremely suitably for pH measurement of cell culture solutions.
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Abstract
Description
また、近年行われている人や動物の細胞や臓器の一部を培養して再生し、病気の治療を行う再生医療においても、細胞を培養する際には、培養状態の管理のため、細胞培養液のpH測定が行われる。この細胞培養におけるpH測定において、細胞培養液を攪拌する際に、細胞が電極に当たるとストレスが生じて細胞培養に悪影響を及ぼすことから、pH電極としては、細胞にストレスを与えない構造が望まれている。
2・・・比較電極
4・・・イオン選択性電極
6・・・複合電極
10・・・容器
12・・・挿入孔
14・・・収容体
15・・・収容体本体
18・・・先端面
36・・・蓋部
38・・・突出部
42・・・環状部材
72・・・変換装置
74・・・表示装置
200、300・・・測定システム
400・・・液体分析計
比較電極2は、図3に示すように、比較電極2用の内部極Rと、内部極Rを収容する収容体14と、内部極Rに接触するように収容体14に充填される比較電極2用のゲル状又は液状の内部液16と、収容体14の先端面18に設けられるとともに内部液16を介して内部極Rと電気的に接続され、試料液8の基準電位を検知するセンサ部としての液絡部22とを備えている。
イオン選択性電極4は、図3に示すように、応答膜24を有する支持体26と、支持体26内に充填されたイオン選択性電極4用の第2の内部液28と、第2の内部液28に浸漬されるイオン選択性電極4用の第2の内部極Mとから構成されており、応答膜24が支持体26の先端面30に設けられ、この応答膜24によって試料液8のイオン濃度に応じた電位が測定される。
具体的には、複合電極6は、前記支持体26を取り囲むように前記収容体14を設けてなり、イオン選択性電極4と比較電極2とを一体的に有するものであり、本実施形態では、回転体形状をなす。
前記挿入孔12は、容器10の一部として形成されるとともに複合電極600が取り付けられる被取付部Xの内部空間である。本実施形態の被取付部Xは、容器10の側壁から外側に向かって突出した円筒状をなすものであり、該被取付部Xの内周面X1に囲まれた空間が、前記挿入孔12として形成されている。ここでは、前記取付部Xの内周面X1は、容器10の外側から内側に向かって徐々に縮径するテーパ状をなす。
なお、上述した構成により、容器10の内壁面32には、前記被取付部Xの内周面X1によって段部が形成されている。
前記位置決め面Yは、容器10に形成された前記挿入方向と反対を向く外向き面に当接する面であり、収容体14が前記挿入孔12に挿入されることにより、前記外向き面から前記挿入方向と反対向きの力を受ける面である。
本実施形態では、上述した被取付部Xの内周面X1及び前記被取付部Xの外端面X2が前記外向き面として形成されており、これらの外向き面それぞれに当接する収容体本体15の外周面Y1及び突出部38の内向き面Y2が前記位置決め面Yである。
より詳細には、前記外周面Y1の全体が前記内周面X1の全体に密接するとともに、前記内向き面Y2が前記外端面X2に当接することにより、収容体14が取付位置に位置決めされる。
なお、前記収容体本体15の外周面Y1は、前記被取付部Xの内周面X1に対応した形状をなし、本実施形態では、後端から先端に向かって徐々に縮径するテーパ状をなす。
ここでいう略面一とは、必ずしも前記先端面18、30と前記内壁面32とが厳密に面一となっている状態に限られず、試料液8の内容物(例えば細胞培養液たる試料液8に含まれる細胞等)が上述した段部や収容体14に引っ掛からない程度に、前記先端面18、30と前記内壁面32とが挿入方向に僅かにずれている状態も含まれる。
より具体的には、前記収容体14が取付位置に取り付けられることにより、収容体14の先端面18、30と、容器10の内壁面32とのずれが5mm以内であることが好ましい。これならば、上述した段部に試料液8の内容物が引っ掛かかりにくくなり、前記先端面18、30を汚れにくくすることができる。また、本実施形態のように、試料液が例えば細胞や微生物等の生体組織を培養する場合、前記ずれが2mm以内であることがより好ましい。何故ならば、前記ずれが2mmより大きいと、細胞や微生物等の培養を開始してから数週間で上述した段部に細胞や微生物等が溜まってしまうという問題が生じるからである。そして、段部に細胞が溜まってしまうと、その溜まった細胞が核となって成長し、目的物質でないものに分化する可能性やコンタミネーションが発生する可能性があるうえ、収率が非常に低下するという問題も生じる。
このような液体分析計400の複合電極6を容器10の挿入孔12にセットすることによって、細胞培養を行いながら、細胞にストレスを与えずに細胞培養液のpH測定を行うことができる。
具体的には、例えば蓋部36が突出部38を有していない場合、収容体14は、位置決め面Yとして、前記収容体本体15の外周面Y1のみを有したものである。
一方、前記被取付部Xの内周面X1及び前記収容体本体15の外周面Y1にテーパが形成されておらず、前記外周面Y1に挿入方向と反対向きの力が作用しない場合、収容体14は、位置決め面Yとして、突出部38の内向き面Y2のみを有したものである。
もちろん、収容体14は、前記実施形態における2つの位置決め面Yとは別に、1又は複数の位置決め面をさらに有するものでも構わない。
上述した場合、位置決め面Yの一部又は全部と被取付部Xとは、同じ材料から形成されていることが好ましく、具体的には例えばポリエチレン等の樹脂から形成されたものが良い。
より具体的な実施態様としては、収容体本体15の外周面Y1及び被取付部Xの内周面X1と、突出部38の内向き面Y2及び被取付部Xの外端面X2との一方又は両方が、熱溶着によって固定されている構成が挙げられる。
具体的には、収容体14の先端面18、30と容器10の内壁面32とが同一面状となる取付位置において、上述した摩擦力が生じるように、収容体本体15の外周面Y1及び被取付部Xの内周面X1に摩擦面が形成されていれば良い。
このようにして、収容体14が取付位置に取り付けられた状態で、該収容体14のさらに外側から例えばゴム製のキャップ部材を嵌め込むことにより、収容体14を前記取付位置に確実に固定することができる。
これならば、上述した摩擦面を形成する必要がなく、収容体14を容易に取付位置に固定することができる。
このような測定システム300においても、複合電極6の先端面18、30が容器62の内壁面と略面一になるようにされているので、洗浄の際には、容器62内を洗浄しやすく、清潔に保つことができる。また、細胞培養液の攪拌の際に、細胞が複合電極6に当たってストレスを受け、細胞培養に悪影響を及ぼすことを未然に防ぐことができる。
なお、測定システム300においても、電極100が上述した比較電極2又はイオン選択性電極4であってもよい。
例えば、種々のプロセスに用いられる配管に挿入孔を設けて、該挿入孔に本発明にかかる電極を挿入すれば、前記配管を流れる排水や上水等のpH等を測定することができる。
また、シャーレに収容された培地で微生物等の生物組織を培養する場合、前記シャーレの側壁に挿入孔を設け、該挿入孔に本発明にかかる電極を挿入することで、顕微鏡による微生物等の観察を邪魔することなく、培地のpH等を測定することができる。
Claims (10)
- 測定対象となる試料液が入れられる容器に設けられた挿入孔に挿入される電極であって、
内部極と、
前記内部極を収容する収容体と、
前記内部極に接触するように前記収容体に充填されるゲル状又は液状の内部液と、
前記内部液を介して前記内部極と電気的に接続され、前記試料液の電位を検知するように前記収容体の先端面に設けられたセンサ部と、を具備し、
前記収容体が、前記容器の内壁面と前記先端面とが同一面状となる取付位置に前記収容体を位置決めするための位置決め面を有していることを特徴とする電極。 - 前記位置決め面が、前記収容体を前記挿入孔に挿入する挿入方向と反対を向きの力を受けることにより、前記収容体が前記取付位置に位置決めされることを特徴とする請求項1の電極。
- 前記容器が、前記挿入方向と反対を向く外向き面を有し、
前記位置決め面が、前記外向き面に当接することにより、前記収容体が前記取付位置に位置決めされることを特徴とする請求項2記載の電極。 - 前記収容体が、前記挿入方向と直交する方向に突出する突出部を具備したものであり、
前記突出部に前記位置決め面が形成されていることを特徴とする請求項2記載の電極。 - 前記突出部に外嵌する環状部材が、前記突出部を前記容器に押圧することによって、前記収容体が前記挿入孔に固定されるように構成してあることを特徴とする請求項4記載の電極。
- 前記収容体が、前記挿入孔に密接して挿入されていることを特徴とする請求項1記載の電極。
- 前記内部極及び前記内部液が、前記試料液の基準電位を測定する比較電極用の内部極及び内部液であり、前記センサ部が液絡部であることを特徴とする請求項1記載の電極。
- 前記内部極及び前記内部液が、前記試料液の電位を測定するイオン選択性電極用の内部極及び内部液であり、前記センサ部が応答膜であることを特徴とする請求項1記載の電極。
- 応答膜を有する支持体と、
前記支持体内に充填された第2の内部液と、
前記第2の内部液に浸漬される第2の内部極と、から構成されるイオン選択性電極と、
請求項7記載の電極と、を備えた複合電極であって、
前記応答膜が前記支持体の先端面に設けられ、
前記応答膜と前記液絡部とが同一平面状に設けられていることを特徴とする複合電極。 - 請求項9記載の複合電極と、
前記複合電極からの測定情報を所望の情報に変換する変換装置と、
前記変換装置によって変換された情報を表示する表示装置と、を備えた液体分析計。
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US15/129,272 US10036718B2 (en) | 2014-03-31 | 2015-03-31 | Electrode, composite electrode, and analyzer for analyzing liquid |
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US10036718B2 (en) | 2018-07-31 |
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US20170102351A1 (en) | 2017-04-13 |
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EP3128319A1 (en) | 2017-02-08 |
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