JP2011188830A - Culture vessel - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a culture vessel reducing intrusion of sundry germs and impurities and/or the like into a well in such a situation as to be installed with necessary electrodes such as a paired electrode and reference electrode. <P>SOLUTION: The culture vessel includes: such a vessel body 2 in which an inner space A open atop thereof is formed and a well 4 is housed in such a way that a cylinder 32 is erected on the top surface of a plate 33 with an action electrode 31 formed atop thereof; a lid 3 fitted detachably on the top of the vessel body 2 and closing the vessel body 2; a columnar electrode 5 penetrating the lid 3, fitted onto the lid 3 in such a state that the top end is exposed onto the top surface of the lid 3, situated in the vessel body 2 in such a state that the lid 3 is fitted onto the vessel body 2, and situated outside the well 4, with the bottom end being in contact with the action electrode 31; and a reference electrode 6 and a paired electrode 7 penetrating the lid 3, each fitted onto the lid 3 in such a state that the respective top ends are exposed onto the top of the lid 3, with the respective bottom ends being intruded into the well 4 in such a state that the lid 3 is fitted onto the vessel body 2. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a culture container for culturing cells, tissues, etc., and more particularly to a culture container used by inserting an electrode for supplying electricity to cells, tissues, etc.

  Containers disclosed in the following patent documents are known as this type of culture container. This container includes a cloning ring composed of a cylindrical glass ring, and an ITO electrode (working electrode) composed of a glass plate having ITO (indium-tin oxide) deposited on the entire surface of the container. An impedance measuring container is constructed by adhering and fixing one end of the cloning ring with an epoxy adhesive at the center of the vapor deposition surface. In this container, a linear platinum wire is connected as an extraction electrode to the ITO deposition surface of the ITO electrode, and the counter electrode (counter electrode) and reference are placed in the container from the other end side (opening end side) of the cloning ring. An electrode is inserted.

Japanese Patent Laying-Open No. 2005-80522 (page 4-5, FIG. 2)

  However, this conventional culture vessel has the following problems to be improved. That is, in this culture vessel, since the counter electrode (counter electrode) and the reference electrode can be inserted, the upper end (the other end portion) of the cloning ring constituting the vessel is open, so that measurement is in progress. In addition, there is a problem that germs, dust, and the like enter the container from the outside of the container through the opening and the culture bodies such as cells and tissues may be contaminated.

  The present invention has been made in order to improve such a problem, and provides a culture vessel that can reduce invasion of germs and dust into the inside in a state where necessary electrodes such as a counter electrode and a reference electrode are mounted. The main purpose.

  In order to achieve the above object, the culture vessel according to the present invention is configured such that a concave portion having an upper opening is formed and a cylindrical body is erected on the upper surface of a plate body in which a working electrode is formed on the upper surface. A container body in which a well is housed in the recess; a lid body detachably attached to the upper portion of the container body; and closing the recess; and an upper end passes through the lid body and an upper end is an upper surface of the lid body Is attached to the lid body in an exposed state, and is located in the recess and outside the well when the lid body is attached to the container body, and the lower end is electrically connected to the working electrode. In a state where the columnar electrode is in contact with the lid body and attached to the lid body with the upper end exposed on the upper surface of the lid body and the lid body is mounted on the container body The lower end enters the well A columnar reference electrode is attached to the lid in a state of penetrating the lid and the upper end is exposed on the upper surface of the lid, and the lower end is attached to the container body in the state where the lid is attached to the container body. A columnar counter electrode that enters the well.

  The culture container according to claim 2 is a container main body formed with a cylindrical body that constitutes the peripheral wall of the well, a plate body that is attached to the lower end of the cylindrical body and constitutes the bottom wall of the well, A lid that is detachably attached to the container body and closes the upper end opening of the well; a working electrode formed on the upper surface of the plate; and the upper end of the lid that passes through the lid A columnar electrode that is attached to the lid in an exposed state on the upper surface, and is located outside the well in a state where the lid is attached to the container body, and a lower end thereof is in electrical contact with the working electrode; The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end enters the well when the lid is attached to the container body. Columnar reference electrode and the lid A columnar counter electrode that penetrates and is attached to the lid body with its upper end exposed on the upper surface of the lid body, and whose lower end enters the well when the lid body is mounted on the container body And.

  In addition, the culture container according to claim 3 has an upper wall in which a plurality of recesses having an upper opening are formed in an aligned state on the upper surface, and a cylindrical body is formed on the upper surface of the plate body on which the working electrode is formed. A container body in which a well constructed in a standing manner is accommodated in each recess, a lid that is detachably attached to the upper surface of the container body, and closes the recess, and penetrates the lid And being attached to the lid body with the upper end exposed on the upper surface of the lid body, and in the corresponding concave portion of the concave portions in a state where the lid body is mounted on the container body, and A plurality of columnar electrodes that are located outside the well housed in the recess and whose lower ends are in electrical contact with the working electrode formed on the plate body constituting the well, and penetrate the lid And the upper end is exposed on the upper surface of the lid. In the state where the lid is attached to the container main body, the lower end of the plurality of columnar pieces that enter into the wells housed in the corresponding recesses of the recesses. A reference electrode is attached to the lid in a state of penetrating the lid and having an upper end exposed on the upper surface of the lid, and the lower end is attached to the container body in the state where the lid is attached to the container body. And a plurality of columnar counter electrodes that enter the wells accommodated in the corresponding ones of the recesses.

  The culture container according to claim 4 is a container main body in which a cylindrical body constituting each peripheral wall of a plurality of wells penetrates the upper wall and is arranged in an aligned state, and the plurality of the plurality of wells. One plate body that is attached to the lower end of the cylindrical body and constitutes the bottom wall of each well, and a lid body that is detachably attached to the container body and closes the upper end openings of the plurality of cylindrical bodies. And a working electrode formed on the upper surface of the plate, and the lid is attached to the lid with the upper end exposed on the upper surface of the lid, and the lid is attached to the container body. 1 or two or more columnar electrodes that are located outside the well and are in electrical contact with the working electrode in a state of being attached to the well, and the upper end passes through the lid, and the upper end is the upper surface of the lid And is attached to the lid body in a state where the lid body is exposed. A plurality of columnar reference electrodes whose lower ends enter the corresponding wells of the respective wells in a state where they are mounted on the main body, and a state where the upper ends are exposed on the upper surface of the lids. And a columnar counter electrode having a lower end that enters the corresponding well of the wells when the lid is attached to the container body.

  The culture container according to claim 5 is the culture container according to any one of claims 1 to 4, wherein the columnar electrode is attached to the lid body so as to be movable back and forth with a predetermined stroke. A biasing member is provided on the body and constantly biases the columnar electrode from the lower end side toward the upper end side of the columnar electrode.

  According to the culture container of claim 1 and the culture container of claim 3, the columnar electrode that is in electrical contact with the working electrode and the lower end are respectively in the well with respect to the container body in which the well is accommodated in the recess. By mounting the lid body to which the reference electrode and the counter electrode to be attached are attached and closing the concave portion formed in the container body, the columnar shape attached to the lid body while closing the concave portion with the lid body An electrode, a reference electrode and a counter electrode can be used to measure the impedance of a culture cultured in a well. Therefore, according to this culture container, invasion of germs and garbage from the outside of the culture container into the culture container 1 is greatly reduced by attaching the lid, and contamination of the culture in the well due to the germs and the like is reduced. The impedance of the culture can be measured with sufficient avoidance.

  According to the culture container according to claim 2 and the culture container according to claim 4, in a state where the lid is attached to the container body, the well is attached from the outside of the culture container by the amount that the upper end of the well is in close contact with the lower surface of the lid. As a result of being able to further reduce the invasion of bacteria and dust into the well than the above-described culture container, it is possible to more reliably avoid the situation where the culture in the well is contaminated by bacteria and the like.

  According to the culture container of claim 5, the columnar electrode that needs to be brought into contact with the working electrode is attached to the lid body so as to be movable back and forth in a predetermined stroke, and the biasing member disposed on the lid body With the configuration in which the columnar electrode is constantly biased from the lower end side toward the upper end side, the container body is closed with a lid to reduce invasion of germs and garbage into the culture container, for example, a probe pin The columnar electrode can be brought into contact with the working electrode without fail by pushing the columnar electrode into the culture vessel.

It is sectional drawing for demonstrating the structure of the culture container 1 in the state before mounting | wearing with the cover body 3. FIG. It is sectional drawing for demonstrating the structure of the culture container 1 in the state with which the cover body 3 was mounted | worn. It is a block diagram of the impedance measuring apparatus 41 using the culture container 1. FIG. It is sectional drawing for demonstrating the structure of other culture container 1A in the state before mounting | wearing with the cover body 3. FIG. It is sectional drawing for demonstrating the structure of 1 A of culture containers in the state which attached the cover body 3 (The columnar electrode 5 and the working electrode 31 are a non-contact state). It is sectional drawing for demonstrating the structure of 1 A of culture containers in the state which attached the cover body (The columnar electrode 5 and the working electrode 31 are a contact state). It is a perspective view of culture container 1B. It is a disassembled perspective view of culture container 1C.

  Hereinafter, embodiments of a culture vessel and an impedance measuring apparatus including the culture vessel will be described with reference to the accompanying drawings.

  First, the configuration of the culture vessel 1 will be described with reference to FIGS.

  As shown in FIGS. 1 and 2, the culture container 1 includes a container body 2, a lid 3, a well 4, a columnar electrode 5, a reference electrode 6, a counter electrode 7, and a biasing member 8.

  The container body 2 is composed of a cylindrical body (cylindrical body or rectangular cylindrical body) whose lower portion is closed using an insulating synthetic resin material. With this configuration, in the container main body 2, the internal space A surrounded by the side wall 11 and the bottom wall 12 is formed as a concave portion whose top is open. For example, as shown in FIG. 1, the lid body 3 is formed in a thin dish shape with an upper wall 21 and a side wall 22 erected around the lid body 3 using an insulating synthetic resin material. As shown in FIG. 2, it is detachably attached to the upper part of the container body 2. Further, an insertion hole 23 for the columnar electrode 5, an insertion hole 24 for the reference electrode 6, and an insertion hole 25 for the counter electrode 7 are formed in the upper wall 21 of the lid 3. In addition, around the insertion hole 23 on the lower surface of the upper wall 21, a cylindrical protrusion that communicates with the insertion hole 23 and the internal space (in this example, a cylindrical protrusion is used as an example. 26 is formed in a state of standing upright to the upper wall 21.

  As shown in FIGS. 1 and 2, the cylindrical protrusion 26 is mounted with the columnar electrode 5 penetrating therethrough, and a coil spring (hereinafter also referred to as “spring 8”) as the biasing member 8 is mounted on the columnar electrode. 5 is housed in a state of being externally fitted. In this example, the inner diameter of the upper region B1 (region directly communicating with the insertion hole 23) B1 in the cylindrical protrusion 26 is the same as the inner diameter of the insertion hole 23 and is larger than the outer diameter of the spring 8. It has a large diameter and is slightly larger than the outer diameter of the medium-diameter cylindrical portion 5b so that the medium-diameter cylindrical portion 5b in the columnar electrode 5 described later can be slidably inserted. Further, the inner diameter of the lower region B2 of the cylindrical projection 26 is such that the columnar electrode 5 can be guided to move forward and backward with little backlash along its axial direction (vertical direction), that is, the small-diameter cylindrical portion 5a. Is formed to have a slightly larger diameter than the diameter of the small-diameter cylindrical portion 5a so as to move while sliding in the lower region B2.

  The well 4 is configured such that a cylindrical body 32 is erected on the surface (upper surface) of the working electrode 31. Specifically, the working electrode 31 functioning as the bottom wall of the well 4 is configured by, for example, an ITO (indium-tin oxide) film deposited on the entire surface (upper surface) of a plate 33 such as a glass plate. Yes. As an example, the cylindrical body 32 that functions as a side wall of the well 4 is configured by a cloning ring configured by a cylindrical glass ring.

  The columnar electrode 5 has a long small diameter cylindrical portion 5a with a lower end formed in a spire shape, a medium diameter cylindrical portion 5b formed at the upper end of the small diameter cylindrical portion 5a, and a large diameter formed at the upper end of the medium diameter cylindrical portion 5b. A stopper pin 5d is provided so as to protrude from the outer peripheral surface of the cylindrical portion 5c and the small-diameter cylindrical portion 5a, and each axis of the small-diameter cylindrical portion 5a, the medium-diameter cylindrical portion 5b, and the large-diameter cylindrical portion 5c is configured to coincide. Yes. In this case, the small-diameter column portion 5 a is formed so that its outer diameter is slightly smaller than the inner diameter of the lower region B <b> 2 in the cylindrical protrusion 26. The medium-diameter column portion 5b has an outer diameter larger than that of the small-diameter column portion 5a, and is slightly smaller than the inner diameter of the upper region B1 in the cylindrical protrusion 26. The length of the medium diameter cylindrical portion 5b is defined to be longer than the distance L described later. The large diameter cylindrical portion 5 c is formed in a flat shape whose outer diameter is larger than the outer diameter of the medium diameter cylindrical portion 5 b and the inner diameter of the insertion hole 23.

  As an example, the columnar electrode 5 is attached to the lid 3 by the following procedure. First, the stopper pin 5d is removed from the outer peripheral surface of the small diameter cylindrical portion 5a, and in this state, the spring 8 is externally fitted to the small diameter cylindrical portion 5a from the tip side. Next, the columnar electrode 5 is inserted into the insertion hole 23 from the distal end side of the small diameter cylindrical portion 5 a and pushed into (inserted into) the insertion hole 23, whereby the distal end side of the small diameter cylindrical portion 5 a is lowered from the cylindrical protrusion 26. To protrude. At this time, the spring 8 externally fitted to the small-diameter cylindrical portion 5 a is inserted into the upper region B 1 of the cylindrical protrusion 26, and the lower end side of the spring 8 is lower than the upper region B 1 on the inner peripheral surface of the cylindrical protrusion 26. It abuts on the step formed at the boundary with the side region B2. For this reason, when the columnar electrode 5 is further inserted into the insertion hole 23, the spring 8 is pushed and shrunk. As a result, an upward biasing force acts on the columnar electrode 5 from the spring 8, but against this biasing force. The lower end side of the medium diameter cylindrical part 5 b of the columnar electrode 5 slightly enters the insertion hole 23 and pushes the columnar electrode 5 to a position where the large diameter cylindrical part 5 c contacts the upper wall 21 of the lid 3. Finally, in this state, the stopper pin 5d is attached to the outer peripheral surface of the small diameter cylindrical portion 5a protruding from the lower end of the cylindrical protrusion 26 (for example, an attachment hole is formed on the outer peripheral surface of the small diameter cylindrical portion 5a, and this To be inserted into the hole).

  In this state, when the pushing to the columnar electrode 5 is released, the columnar electrode 5 is moved upward by the urging force from the spring 8, and when the stopper pin 5 d comes into contact with the lower end of the cylindrical protrusion 26, the spring 8 is moved. The movement is stopped with the urging force applied from. In this stopped state, as shown in FIG. 1, the columnar electrode 5 has a lower end side of the medium-diameter columnar portion 5 b slightly entering the insertion hole 23, and the large-diameter columnar portion 5 c extends from the upper wall 21 of the lid 3. It is in a position separated by a distance L. Therefore, the columnar electrode 5 is inserted through the insertion hole 23 and the cylindrical protrusion 26, thereby penetrating the lid body 3 and at a right angle with respect to the lid body 3. The upper end of the cylindrical protrusion 26 is the upper limit, and the lower end is the position where the large-diameter column portion 5c is in contact with the lid 3. The large-diameter column portion 5c (upper end) is the lid. Attached to the lid 3 in an exposed state on the upper surface of the body 3. In addition, the protrusion length from the lower end of the cylindrical protrusion 26 of the small-diameter columnar portion 5a in a state where the stopper pin 5d is in contact with the lower end of the cylindrical protrusion 26 is such that the lid 3 is placed on the container body 2 as shown in FIG. The tip (lower end) of the small diameter cylindrical portion 5a is not in contact with the working electrode 31, and is separated from the working electrode 31 within the stroke range (distance L) of the columnar electrode 5 (in other words, The length of the columnar electrode 5 is defined such that the tip of the small-diameter cylindrical portion 5a comes into contact with the working electrode 31).

  The reference electrode 6 includes a long small-diameter cylindrical portion 6a and a large-diameter cylindrical portion 6b formed at the upper end of the small-diameter cylindrical portion 6a. The reference electrode 6 is a columnar electrode in which the axes of the small-diameter cylindrical portion 6a and the large-diameter cylindrical portion 6b coincide. It is configured. In this case, the small-diameter cylindrical portion 6 a is formed so that its outer diameter is slightly larger than the inner diameter of the insertion hole 24. The large diameter cylindrical portion 6b is formed in a flat shape whose outer diameter is larger than the outer diameter of the small diameter cylindrical portion 6a. The reference electrode 6 is inserted into the insertion hole 24 from the distal end side of the small-diameter cylindrical portion 6a until the large-diameter cylindrical portion 6b contacts the upper wall 21 of the lid 3, thereby penetrating the lid 3. The large-diameter cylindrical portion 6b (upper end) is attached to the lid body 3 in a state of being fixed to the lid body 3 at a right angle and being exposed on the upper surface of the lid body 3.

  The counter electrode 7 includes a long small-diameter cylindrical portion 7a and a large-diameter cylindrical portion 7b formed at the upper end of the small-diameter cylindrical portion 7a. The counter-electrode 7 is a columnar electrode in which the axes of the small-diameter cylindrical portion 7a and the large-diameter cylindrical portion 7b coincide. It is configured. In this case, the small-diameter cylindrical portion 7 a is formed such that its outer diameter is slightly larger than the inner diameter of the insertion hole 25. The large diameter cylindrical portion 7b is formed in a flat shape whose outer diameter is larger than the outer diameter of the small diameter cylindrical portion 7a. The counter electrode 7 is inserted into the insertion hole 25 from the distal end side of the small diameter cylindrical portion 7 a until the large diameter cylindrical portion 7 b comes into contact with the upper wall 21 of the lid 3, thereby penetrating the lid 3. The large-diameter cylindrical portion 7b (upper end) is attached to the lid 3 in a state of being fixed to the lid 3 at a right angle and being exposed on the upper surface of the lid 3. Each protrusion length of the reference electrode 6 and the counter electrode 7 from the lower surface of the upper wall 21 in the lid 3 is such that the small-diameter cylindrical portion 6a in the state where the lid 3 is mounted on the container body 2 as shown in FIG. , 7a is defined such that the tip (lower end) enters the cylindrical body 32 of the well 4 and does not contact the working electrode 31. The reference electrode 6 and the counter electrode 7 may be attached to the insertion holes 24 and 25 by adopting a structure in which the reference electrode 6 and the counter electrode 7 are attached by bonding instead of press-fitting.

  Next, the structure of the impedance measuring apparatus 41 provided with the culture container 1 is demonstrated with reference to FIG.

  The impedance measuring device 41 includes a culture vessel 1, an incubator 42, a fixing jig 43, a probe head 44, a measuring device 45, and a computer 46, and the culture vessel 1 fixed to the fixing jig 43 disposed in the incubator 42. The impedance Z of the inner culture body (for example, the culture body (not shown) cultured in the liquid medium 101) was brought into contact with the columnar electrode 5, the reference electrode 6 and the counter electrode 7 of the culture vessel 1. Measurement is possible using a measuring device 45 and a computer 46 disposed outside the incubator 42 via the probe head 44.

  As an example, the fixing jig 43 includes a mounting table 43a installed on the bottom surface of the incubator 42 and a plurality of columns 43b standing on the upper surface of the mounting table 43a. In the fixing jig 43, the culture vessel 1 is placed at a predetermined position on the mounting table 43a, and probe pins 44, which will be described later, are provided on the upper ends of the respective columns 43b. And it attaches so that it may contact with respect to the counter electrode 7 from upper direction. Thereby, the culture container 1 is fixed to the fixing jig 43 while being sandwiched between the mounting table 43a and the probe head 44.

  The probe head 44 is a base 44a and a columnar electrode in the lid 3 of the culture vessel 1 placed at a predetermined position on the mounting base 43a on one surface (the lower surface in FIG. 3) side of the base 44a. 5, three probe pins 44b, 44c, and 44d are provided so as to project corresponding to the positions of the reference electrode 6 and the counter electrode 7, respectively. The probe pins 44b, 44c, and 44d are attached to the base 44a so as to be able to advance and retract (protrude into) by a spring mechanism (not shown) provided on the base 44a. Further, the urging force of the spring mechanism with respect to each probe pin 44b, 44c, 44d is defined to be stronger than the urging force of the spring 8 disposed on the lid 3.

  The measuring device 45 includes an AC constant voltage source and an ammeter (both not shown). In this case, the AC constant voltage source is connected to the probe pin 44b of the probe head 44 disposed in the incubator 42 via the cable 51a, and is connected to the probe pin 44d via the cable 51c. An AC constant voltage Vm is applied between the pins 44b and 44d. The AC constant voltage source outputs waveform data Dv of the AC constant voltage Vm to the computer 46. On the other hand, the ammeter is connected to the probe pin 44b via the cable 51a and is connected to the probe pin 44c via the cable 51b, and each probe is caused by the application of the AC constant voltage Vm by the AC constant voltage source. The alternating current Im flowing between the pins 44b and 44c is detected, and waveform data Di indicating the amplitude of the alternating current Im is output to the computer 46.

  The computer 46 captures the waveform data Dv and the waveform data Di for a predetermined time (for example, several cycles of the AC constant voltage Vm) and stores it in the internal memory, and stores the stored waveform data Dv and waveform data Di. Each of the AC constant voltage Vm and the AC current Im such as the phase difference between the AC constant voltage Vm and the AC current Im, and the amplitudes (instantaneous values at the same time) of the AC constant voltage Vm and the AC current Im. The characteristic value is calculated, and based on the calculated characteristic value, the impedance Z between the probe pins 44b and 44c is calculated, stored in the internal memory, and displayed on the internal display device. The impedance Z calculated by the computer 46 is a state in which the probe pins 44b, 44c and 44d are in contact with the columnar electrode 5, the reference electrode 6 and the counter electrode 7 of the culture vessel 1, respectively, as shown in FIG. Is the impedance of the culture cultured in the well 4 in the culture vessel 1.

  Then, the measurement operation | movement about the impedance Z of the culture body by the impedance measuring apparatus 41 is demonstrated.

  First, prior to measuring the impedance Z, the culture is cultured in the well 4 of the culture vessel 1. Specifically, the lid 3 is removed from the container body 2 and the medium 101 is placed in the well 4. Next, the culture medium (cells or tissue) is seeded on the medium 101, and subsequently, as shown in FIG. 2, the small diameter cylindrical portions 6a and 7a of the reference electrode 6 and the counter electrode 7 enter the well 4, The lid 3 is attached to the container body 2 so that the lower end side is immersed in the culture medium 101 and the columnar electrode 5 is located outside the well 4. Thereby, the upper end opening of the container body 2 is closed by the lid 3.

  In this culture vessel 1, although the insertion hole 23, the insertion hole 24 and the insertion hole 25 for attaching the columnar electrode 5, the reference electrode 6 and the counter electrode 7 to the lid body 3 are formed, the insertion hole 24 and the insertion hole 25. Is closed by press-fitting the reference electrode 6 and the counter electrode 7, and for the insertion hole 23, each inner diameter of the upper region B1 of the insertion hole 23 and the cylindrical projection 26 directly communicating with the insertion hole 23 is the medium-diameter column portion 5b. Is defined as an inner diameter that advances and retracts in the sliding state, and an inner diameter of the lower region B2 of the cylindrical protrusion 26 is defined as an inner diameter that allows the small-diameter column portion 5a to advance and retract in the sliding state. 5 is almost close to blocking. For this reason, in the culture container 1, in the state where the lid body 3 is attached to the container body 2, the invasion of germs and dust from the outside of the culture container 1 into the culture container 1 is greatly reduced, and the well 4 The situation where the inside culture body is contaminated by various bacteria is sufficiently avoided.

  Next, the culture vessel 1 is placed on a fixing jig 43 provided in the incubator 42, and the probe pins 44 b, 44 c and 44 d of the probe head 44 are brought into contact with the corresponding columnar electrode 5, reference electrode 6 and counter electrode 7. At the same time, the probe head 44 is fixed to the fixing jig 43. At this time, the probe pins 44c and 44d that are in contact with the reference electrode 6 and the counter electrode 7 fixedly attached to the lid 3 are brought into contact with the reference electrode 6 and the counter electrode 7 in the base 44a. It is in a slightly depressed state. For this reason, the reference electrode 6 and the probe pin 44c, and the counter electrode 7 and the probe pin 44d are maintained in a reliably contacted state. For the columnar electrode 5 attached to the lid 3 so as to be able to move forward and backward, the urging force of the spring mechanism in the probe head 44 is stronger than the urging force of the spring 8 in the lid 3. Due to the contact with 44b, the tip of the small-diameter cylindrical portion 5a is caused to enter the culture vessel 1 until it contacts the working electrode 31 constituting the well 4. For this reason, the columnar electrode 5 and the probe pin 44b are also maintained in a reliable contact state.

  Subsequently, the measurement device 45 starts to operate, starts application of the AC constant voltage Vm between the probe pins 44b and 44d via the cables 51a and 51c, and outputs the waveform data Dv of the AC constant voltage Vm to the computer 46. Output to. The measuring device 45 detects the alternating current Im flowing between the probe pins 44b and 44c via the cables 51a and 51b, and outputs waveform data Di indicating the amplitude of the alternating current Im to the computer 46.

  Next, the computer 46 calculates the impedance Z between the probe pins 44b and 44c, that is, the impedance Z of the culture cultured in the well 4 in the culture vessel 1, based on the waveform data Dv and the waveform data Di. And stored in the internal memory and displayed on the internal display device. Thereby, the measurement of the impedance Z of the culture by the impedance measuring device 41 is completed.

  Thus, according to the culture vessel 1, the columnar electrode 5 that makes electrical contact with the working electrode 31 that functions as the bottom wall of the well 4 and the lower end of the vessel body 2 in which the well 4 is housed are respectively provided. By mounting the lid body 3 to which the reference electrode 6 and the counter electrode 7 entering the well 4 are attached to close the container body 2, the lid body 3 closes the container body 2 while closing the lid. The impedance Z of the culture cultured in the well 4 can be measured using the columnar electrode 5, the reference electrode 6, and the counter electrode 7 attached to the body 3. Therefore, according to this culture container 1, the invasion of germs and garbage from the outside of the culture container 1 into the culture container 1 is greatly reduced by the attachment of the lid 3, so that the germs of the culture body in the well 4 can be reduced. The impedance Z can be measured while sufficiently avoiding contamination due to the like.

  Moreover, according to this culture container 1, the columnar electrode 5 that needs to be brought into contact with the working electrode 31 is attached to the lid 3 so as to be movable back and forth in a predetermined stroke, and the spring 8 disposed on the lid 3 is provided. The column body 5 is always biased from the lower end side toward the upper end side (toward the upper limit direction of the stroke), thereby closing the container body 2 with the lid 3 and culturing containers for germs and garbage. The columnar electrode 5 can be reliably brought into contact with the working electrode 31 by pushing the columnar electrode 5 into the culture vessel 1 by the probe pin 44b of the probe head 44 while reducing intrusion into the chamber 1.

  Further, according to the culture vessel 1, the inner diameter of the upper region B1 and the middle-diameter column portion of the columnar electrode 5 are slid so that the middle-diameter column portion 5b of the columnar electrode 5 slides in the upper region B1 of the cylindrical protrusion 26. The outer diameter of 5b and the inner diameter of the lower region B1 and the smaller diameter cylindrical portion of the columnar electrode 5 so that the smaller diameter cylindrical portion 5c of the columnar electrode 5 slides in the lower region B2 of the cylindrical protrusion 26. By defining the outer diameter of 5c, even in the configuration in which the columnar electrode 5 is attached to the lid 3 so as to be movable back and forth, the internal space of the insertion hole 23 and the cylindrical projection 26 is blocked by the columnar electrode 5, It is possible to reduce the intrusion of garbage into the culture container 1.

  In the culture container 1 described above, a configuration in which the upper end opening of the container body 2 containing the well 4 is closed by the lid 3 is employed. However, as in the culture container 1A shown in FIGS. A configuration in which the upper end opening of 4A is directly closed by the lid 3 can also be adopted. Hereinafter, the configuration of the culture vessel 1A will be described. In addition, about the structure same as the culture container 1, the same code | symbol is attached | subjected and the overlapping description is abbreviate | omitted.

  As shown in FIGS. 4, 5, and 6, the culture container 1 </ b> A includes a container body 2 </ b> A, a lid 3, a well 4 </ b> A, a columnar electrode 5, a reference electrode 6, a counter electrode 7, and a biasing member 8. The lid 3, and the columnar electrode 5, the reference electrode 6, the counter electrode 7 and the biasing member 8 (that is, the assembly on the lid 3 side) attached to the lid 3 are the same as the culture vessel 1, The configuration of the container body 2A and the well 4A will be mainly described.

  As shown in FIGS. 4 and 5, the container main body 2 </ b> A includes a cylindrical side wall 61, an upper wall 62, and a cylindrical body 63. Specifically, an upper wall 62 is formed at the upper portion of the side wall 61, and the upper wall 62 penetrates the upper wall 62 and the length direction (vertical direction in the figure) is the length direction of the side wall 61. A cylindrical body 63 is formed so as to be in parallel with each other. Moreover, the cylindrical body 63 is comprised by the cylinder (cylindrical body or a square cylinder) which an upper end and a lower end open. The cylindrical body 63 is formed so that the upper end is flush with the upper end of the side wall 61 and the lower end is positioned above the lower end of the side wall 61 (the lower end is positioned inside the side wall 61). Yes. The upper wall 62 has an insertion hole 64 through which the cylindrical protrusion 26 of the lid 3 is inserted.

  The well 4A includes a cylindrical body 63 of the container body 2A and a plate body 33 having a working electrode 31 formed on the upper surface, and the cylindrical body 32 is erected on the surface (upper surface) of the working electrode 31. ing. That is, the cylindrical body 63 constitutes the peripheral wall (side wall) of the well 4A, and the working electrode 31 constitutes the bottom wall of the well 4A. In this case, the working electrode 31 is attached to the lower end of the cylindrical body 63 in a state where the ITO 31 a is in close contact with the lower end of the cylindrical body 63. The size of the working electrode 31 is formed so as to extend up to just below the insertion hole 64 when the lid 3 is mounted on the container body 2A.

  In this culture vessel 1A, as shown in FIG. 5, the small-diameter cylindrical portions 6a and 7a of the reference electrode 6 and the counter electrode 7 enter the well 4A, the lower end thereof is immersed in the culture medium 101, and the cylindrical protrusion 26 is formed. When the lid 3 is attached to the container main body 2 so as to be inserted into the insertion hole 64, the upper end of the well 4A (that is, the upper end of the cylindrical body 63) is brought into close contact with the lower surface of the lid 3, so that the well The upper end opening of 4A (that is, the upper end opening of the cylindrical body 63) is closed by the lid 3. Moreover, in this culture container 1A, although the insertion hole 24 and the insertion hole 25 are formed in the site | part which faces the inside of the well 4A in the cover body 3, about the insertion hole 24 and the insertion hole 25, the reference electrode 6 and The counter electrode 7 is blocked by being press-fitted.

  Therefore, according to the culture container 1A, in the state where the lid 3 is attached to the container body 2, the bacteria and dust from the outside of the culture container 1 are as much as the upper end of the well 4A is in close contact with the lower surface of the lid 3. As a result, the invasion into the well 4A can be further reduced as compared with the culture container 1 described above. As a result, the situation where the culture in the well 4A is contaminated with various bacteria can be avoided more reliably.

  Also in the culture vessel 1A, the columnar electrode 5 that needs to be brought into contact with the working electrode 31 is attached to the lid 3 so as to be able to move forward and backward with a predetermined stroke in the same manner as in the culture vessel 1, and the lid 3 employs a configuration in which the column electrode 5 is constantly urged from the lower end side toward the upper end side by the spring 8 disposed in the cylinder 3, so that the well 4 is closed by the lid 3 and the inside of the well 4 such as germs and dust The columnar electrode 5 can be reliably brought into contact with the working electrode 31 by pushing the columnar electrode 5 into the culture vessel 1 by the probe pin 44b of the probe head 44 while reducing the penetration into the working electrode 31.

  In addition, the culture vessels 1 and 1A in which one well 4 and 4A are arranged in one vessel main body 2 and 2A have been described above, but a plurality of wells are arranged in one vessel main body, and a lid body is provided on the container main body. It is also possible to adopt a configuration that simultaneously reduces the invasion of germs and the like into a plurality of wells by mounting the. Hereinafter, culture vessels 1B and 1C employing this configuration will be described with reference to FIGS.

  First, the configuration of the culture vessel 1B will be described with reference to FIG. As will be described later, the culture vessel 1B employs a configuration in which the concave portion of the culture vessel containing the well is closed with a lid (a configuration in which the well is not directly closed with the lid). Since it is similar to the container 1, the same components as those of the culture container 1 are denoted by the same reference numerals and redundant description is omitted.

  The culture container 1B includes a single container body 2B in which a plurality of recesses 73 for accommodating the wells 4 are formed, and a columnar electrode 5 and a reference electrode 6 in each region corresponding to each recess 73 (region C indicated by a one-dot chain line). And one lid 3B in which a set of counter electrodes 7 is disposed.

  Specifically, the container body 2B is disposed as a cylinder (in this example, a square cylinder) as an example, and the upper end of the sidewall 71 covers the upper opening of the sidewall 71. The upper wall 72 is provided. The upper wall 72 is formed with a plurality of (in this example, four as an example) concave portions 73 whose upper portions are open on the upper surface of the upper wall 72 in an aligned state. In each recess 73, the well 4 is accommodated with the lower surface of the working electrode 31 in close contact with the bottom surface of the recess 73.

  As an example, the lid 3B includes a top wall 21B and a side wall 22B standing around the upper wall 21B, is formed in a thin dish shape, and is detachably mounted on the upper portion of the container body 2B. Further, the columnar electrode 5, the reference electrode 6 and the counter electrode 7 are disposed on the upper wall 21B of the lid 3B in a region C corresponding to the position of the upper opening of each recess 73 formed in the container body 2B. Has been. In addition, since the attachment structure to the cover body 3B of the columnar electrode 5, the reference electrode 6, and the counter electrode 7 is the same as the attachment structure of the culture vessel 1, description is abbreviate | omitted.

  With this configuration, in the culture vessel 1B, although not shown, the upper opening of each recess 73 is closed by the lid 3B in a state where the lid 3B is attached to the vessel body 2B. In each recess 73, the columnar electrode 5 attached in the region C corresponding to the recess 73 in the lid 3B is located outside the well 4, and the spring 8 is attached to the recess 73 by the probe pin 44b. When pushed against the force, the lower end of the columnar electrode 5 is in electrical contact with the working electrode 31. In addition, the reference electrode 6 and the counter electrode 7 attached in the region C both enter the well 4, and the lower end side thereof is immersed in the medium 101 accommodated in the well 4.

  Therefore, according to this culture vessel 1B, since the upper opening of each recess 73 is closed by the lid 3B in a state where the lid 3B is attached to the vessel body 2B, various germs and garbage from the outside of the culture vessel 1B. As a result of significantly reducing the intrusion into each well 4, it is possible to sufficiently avoid the situation where the culture in each well 4 is contaminated by various germs.

  Next, the configuration of the culture vessel 1C will be described with reference to FIG. As will be described later, this culture vessel 1C is similar to the culture vessel 1A described above in that it employs a configuration in which the well is directly closed with a lid. Therefore, the same reference numerals are used for the same components as the culture vessel 1A. A duplicate description will be omitted.

  The culture container 1C includes a container body 2C in which a plurality of cylindrical bodies 63 constituting the well 4A are formed, and a reference electrode 6 in each area (area D indicated by a one-dot chain line) corresponding to each cylindrical body 63. And a pair of counter electrodes 7, and one lid 3 </ b> C in which one or more (two in this example, two) columnar electrodes 5 are disposed.

  Specifically, the container body 2 </ b> C is disposed as a cylinder (in this example, a rectangular cylinder) as an example, and an upper portion of the sidewall 81 is disposed so as to cover the upper opening of the sidewall 81. The upper wall 82 is provided. A cylindrical body 63 constituting each peripheral wall of the plurality of wells 4A passes through the upper wall 82 and is arranged in an aligned state on the upper wall 82. The upper end of each cylindrical body 63 is defined to be flush with the upper end of the side wall 81. The upper wall 82 has two insertion holes 64 through which the cylindrical protrusions 26 of the lid 3 are inserted, the same number as the cylindrical protrusions 26. Further, in a state in plan view, the working electrode as a single plate formed in a size including all the cylindrical bodies 63 and all the insertion holes 64 (in this example, a quadrangle matching the planar shape of the side wall 81). 31 is attached and disposed at a position indicated by a two-dot chain line inside the container body 2 </ b> C in a state of being in close contact with the lower ends of all the cylindrical bodies 63. With this configuration, the container body 2 </ b> C has the same number of wells 4 </ b> A as the cylindrical body 63 having the cylindrical body 63 as a peripheral wall and the working electrode 31 as a common bottom wall.

  As an example, the lid 3C includes a top wall 21C and a side wall 22C erected around the top wall 21C, is formed in a thin dish shape, and is detachably attached to the upper portion of the container body 2C. Further, on the upper wall 21C of the lid 3C, a set of the reference electrode 6 and the counter electrode 7 is disposed in a region D corresponding to the position of the upper opening of each cylindrical body 63 formed in the container body 2C. Has been. Further, on the upper wall 21C of the lid 3C, as an example, the columnar electrodes 5 are disposed outside the region D at a ratio of two pairs of the reference electrode 6 and the counter electrode 7 (two in this example in total). Yes. In addition, since the attachment structure to the cover body 3B of the columnar electrode 5, the reference electrode 6, and the counter electrode 7 is the same as the attachment structure of the culture vessel 1, description is abbreviate | omitted.

  With this configuration, in this culture vessel 1C, although not shown, in the state where the lid 3C is mounted on the vessel body 2C, the upper opening of each cylindrical body 63, that is, the upper opening of each well 4A is the lid 3C. It is blocked by. The columnar electrode 5 attached outside the region D in the lid 3C is positioned outside the well 4A and is pushed into the container body 2C against the biasing force of the spring 8 by the probe pin 44b. The lower end of the columnar electrode 5 is in electrical contact with the working electrode 31. On the other hand, the reference electrode 6 and the counter electrode 7 attached in the region D both enter the well 4A, and the lower end side thereof is immersed in the medium 101 accommodated in the well 4A.

  Therefore, according to the culture container 1C, the upper end of each well 4A is in close contact with the lower surface of the lid 3 in the same manner as the culture container 1A in the state where the lid 3C is attached to the container main body 2C. As a result of being able to further reduce the invasion of germs and garbage from the outside of the culture vessel 1C into the well 4A as compared with the culture vessel 1 described above, the culture in each well 4A is contaminated by the germs and the like. Can be avoided more reliably.

  In the culture vessels 1, 1A, 1B, and 1C, a seal member (for example, an O-ring) is disposed between the outer peripheral surface of the columnar electrode 5 and the inner peripheral surfaces of the insertion hole 23 and the cylindrical protrusion 26. It can also be adopted. By adopting this configuration, in particular, in the culture vessels 1 and 1B, it is ensured by the seal member that a germ or the like enters through the insertion hole 23 into which the columnar electrode 5 is inserted and the inside of the cylindrical protrusion 26. Since it can prevent, the situation where the culture body in the well 4 is contaminated by various bacteria etc. can be avoided more reliably. Moreover, about the structure which attaches the columnar electrode 5 to the cover body 3 so that advance / retreat is possible, it can replace with an above-described structure, and can employ | adopt various structures.

  In addition, in the culture containers 1, 1A, 1B, 1C, the columnar electrode 5 is placed in the container body 2 in a state where the cover body 3 (3B, 3C) is mounted on the container body 2 (2A, 2B, 2C). Only after being pushed in, is attached to the lid 3 (3B, 3C) so as to be able to move forward and backward so that the lower end contacts the working electrode 31, and is biased toward the upper end by the spring 8 (biasing member). Although the configuration is adopted, by omitting the spring 8, the column electrode 5 has its own weight and the stroke range when the lid 3 (3B, 3C) is mounted on the container body 2 (2A, 2B, 2C). It is also possible to adopt a configuration in which the lower end thereof contacts the working electrode 31 by moving downward. Also in this configuration, the columnar electrode 5 can be reliably brought into contact with the working electrode 31 by being pushed further downward by the probe pin 44b.

  Although not shown, the columnar electrode may be configured such that at least the lower end side is formed in a leaf spring shape and is press-fitted into the insertion hole 23 formed in the lid and fixed. The columnar electrode in this configuration is not configured to be movable back and forth with respect to the lid, but when the lid is attached to the container body and the lower end abuts against the working electrode 31, the plate spring-like electrode Since the lower end side is elastically deformed, the contact between the lower end and the working electrode 31 is reliably maintained.

1, 1A, 1B, 1C Cultivation vessel 2, 2A, 2B, 2C Vessel body 3, 3B, 3C Lid 4, 4A Well 5 Column electrode 6 Reference electrode 7 Counter electrode 31 Working electrode 32 Tubular body 63 Tubular body 73 Recess

Claims (5)

A container body in which a well formed by a cylindrical body standing on the upper surface of a plate body having a working electrode formed on the upper surface is formed in a recess having an upper opening,
A lid that is detachably attached to the upper portion of the container body and closes the recess;
The lid body is attached to the lid body with the upper end exposed at the upper surface of the lid body, and the lid body is mounted in the container body and is in the recess, and A columnar electrode located outside the well and having a lower end in electrical contact with the working electrode;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end enters the well when the lid is attached to the container body. A columnar reference electrode;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end enters the well when the lid is attached to the container body. A culture vessel comprising a columnar counter electrode. A container body in which a cylindrical body constituting the peripheral wall of the well is formed;
A plate body attached to the lower end of the cylindrical body and constituting the bottom wall of the well;
A lid that is detachably attached to the container body and closes the upper end opening of the well;
A working electrode formed on the upper surface of the plate,
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end is located outside the well when the lid is attached to the container body. A columnar electrode in electrical contact with the working electrode;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end enters the well when the lid is attached to the container body. A columnar reference electrode;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end enters the well when the lid is attached to the container body. A culture vessel comprising a columnar counter electrode. Each of the wells, each of which has a top wall formed with a plurality of concave portions having an upper portion formed in an aligned state on the upper surface and a cylindrical body standing on the upper surface of the plate having a working electrode formed on the upper surface, A container body housed in the recess;
A lid that is detachably attached to the upper surface of the container body and closes the recesses;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the corresponding recess among the recesses when the lid is attached to the container body. A plurality of columnar electrodes that are located inside and outside of the well housed in the recess, and whose lower ends are in electrical contact with the working electrode formed on the plate constituting the well; ,
The lid body is attached to the lid body with the upper end exposed on the upper surface of the lid body, and the lower end of each of the recesses is attached to the container body. A plurality of columnar reference electrodes that enter the wells housed in corresponding recesses;
The lid body is attached to the lid body with the upper end exposed on the upper surface of the lid body, and the lower end of each of the recesses is attached to the container body. A culture vessel comprising a plurality of columnar counter electrodes entering the wells accommodated in corresponding recesses. A cylindrical body constituting each peripheral wall of a plurality of wells on the upper wall, penetrating the upper wall, and a container body disposed in an aligned state,
A single plate attached to the lower ends of the plurality of cylindrical bodies and constituting the bottom wall of each well;
A lid that is detachably attached to the container body and closes upper end openings of the plurality of cylindrical bodies;
A working electrode formed on the upper surface of the plate,
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end is located outside the well when the lid is attached to the container body. One or more columnar electrodes in electrical contact with the working electrode;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end of each well is attached to the container body. A plurality of columnar reference electrodes that enter the corresponding wells;
The lid is attached to the lid with the upper end exposed at the upper surface of the lid, and the lower end of each well is attached to the container body. A culture vessel comprising a columnar counter electrode that enters a corresponding well. The columnar electrode is attached to the lid so as to be movable back and forth with a predetermined stroke,
The culture according to any one of claims 1 to 4, further comprising an urging member that is disposed on the lid and constantly urges the columnar electrode from the lower end side toward the upper end side of the columnar electrode. container.

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