WO2021054103A1 - Dispositif de mesure de résistance électrique, procédé de mesure de résistance électrique, et dispositif de calcul de résistance électrique - Google Patents

Dispositif de mesure de résistance électrique, procédé de mesure de résistance électrique, et dispositif de calcul de résistance électrique Download PDF

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WO2021054103A1
WO2021054103A1 PCT/JP2020/032922 JP2020032922W WO2021054103A1 WO 2021054103 A1 WO2021054103 A1 WO 2021054103A1 JP 2020032922 W JP2020032922 W JP 2020032922W WO 2021054103 A1 WO2021054103 A1 WO 2021054103A1
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electrical resistance
electrode
electric resistance
measuring device
reference electrode
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PCT/JP2020/032922
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English (en)
Japanese (ja)
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正瑛 山▲崎▼
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株式会社Ｓｃｒｅｅｎホールディングス
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Publication of WO2021054103A1 publication Critical patent/WO2021054103A1/fr

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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
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/04—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01R—MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES
    • G01R27/00—Arrangements for measuring resistance, reactance, impedance, or electric characteristics derived therefrom
    • G01R27/02—Measuring real or complex resistance, reactance, impedance, or other two-pole characteristics derived therefrom, e.g. time constant

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  • the present invention relates to a technique for measuring the electrical resistance of a biological sample.
  • Transepithelial electrical resistance (TEER) measurement is known as a method for evaluating the barrier function of the cell layer that forms the membrane structure.
  • a bottomed tubular insert having a bottom surface made of a porous membrane is arranged in the recess of the culture plate, and cells are cultured on the porous membrane.
  • a working electrode for applying a current is arranged inside and outside the insert, and a reference electrode for measuring a potential difference is arranged. Then, the electrical resistance of the cell layer can be obtained by measuring the potential difference generated between the reference electrodes while applying a current between the working electrodes.
  • an AC impedance method in which an AC signal is applied to cells may be adopted. According to the AC impedance method, less damage is given to cells and the measurement time can be shortened as compared with the DC polarization measurement in which a DC waveform is applied. In addition, there is a lot of information obtained from measurements such as frequency response and phase difference.
  • An object of the present invention is to provide a technique for effectively correcting an offset generated in electrical resistance measurement.
  • the first aspect is an electric resistance measuring device for measuring the electric resistance of a biological sample, wherein the first working electrode and the second working electrode, the first reference electrode and the second reference electrode are used.
  • An AC power supply connected to the first acting electrode and the second acting electrode to apply an AC signal having a fixed period, and a voltmeter connected to the first reference electrode and the second reference electrode to measure a potential difference.
  • It includes an electric resistance calculation unit that calculates the electric resistance of the biological sample based on the voltage value obtained by the voltmeter, and the electric resistance calculation unit is a unit from the difference value of two voltage values in the same phase. It is provided with an offset calculation unit that executes a process of calculating the amount of change in the voltage value per hour.
  • the second aspect is the electric resistance measuring device of the first aspect, in which the electric resistance calculation unit subtracts the offset based on the change amount obtained by the offset calculation unit from the voltage value obtained by the voltmeter. To execute.
  • the third aspect is the electric resistance measuring device of the first aspect or the second aspect, and the electric resistance calculation unit sets the time when each of the two voltage values was measured as the first time and the second time.
  • the electric resistance is calculated from the voltage value measured within the period from the first hour to the second hour.
  • the fourth aspect is the electric resistance measuring device according to any one of the first to third aspects, and at least one of the first reference electrode and the second reference electrode is a thin film electrode.
  • the fifth aspect is the electrical resistivity measuring device of the fourth aspect, in which the thin film electrode is composed of either gold (Au), platinum (Pt), or indium tin oxide (ITO).
  • the sixth aspect is an electric resistance measuring method for measuring the electric resistance of a biological sample, which is constant between (a) a first working electrode and a second working electrode arranged with the biological sample sandwiched between them. Obtained by the potential difference measuring step of measuring the potential difference between the first reference electrode and the second reference electrode arranged across the biological sample while applying a periodic AC signal, and (b) the potential difference measuring step. Includes a step of calculating the difference value of two voltage values in the same phase, and (c) a step of calculating the amount of change of the voltage value per unit time from the difference value.
  • an AC signal having a fixed period is applied between the first reference electrode located sandwiching the biological sample and the second acting electrode located on the other side of the biological sample
  • one of the biological samples is applied.
  • An electrical resistance calculation device that calculates the electrical resistance of the biological sample from the potential difference generated between the first reference electrode located on the side and the second reference electrode located on the other side of the biological sample, and has the same phase. It is provided with an offset calculation unit that calculates the amount of change of the voltage value per unit time from the difference value of the two voltage values in.
  • the amount of change in the voltage value per unit time can be obtained from the difference value of the two voltage values in the same phase. This makes it possible to obtain an offset that changes significantly over time. Therefore, the offset generated in the electric resistance measurement can be effectively corrected.
  • the electric resistance of the biological sample can be accurately obtained by subtracting the offset from the measured voltage value.
  • the electric resistance measuring device of the third aspect when the amount of change in the voltage value per unit time is obtained from the amount of change in the voltage value from the first time to the second time, the period from the first time to the second hour.
  • the offset inside can be calculated appropriately.
  • the voltage value during the period from the first hour to the second hour can be corrected with high accuracy, so that the electrical resistance of the biological sample can be accurately obtained from the voltage value during the period.
  • the electric resistance measuring device of the fourth aspect by using the first reference electrode or the second reference electrode as the thin film electrode, it is possible to prevent the first reference electrode or the second reference electrode from being hindered in the electric resistance measurement. ..
  • a printing method can be adopted when producing thin film electrodes of gold, platinum, and ITO. Further, since the influence of the offset due to the electric double layer caused by using the thin film electrode of gold, platinum or ITO can be removed by the correction process, the electric resistance of the biological sample can be accurately performed.
  • the amount of change in the voltage value per unit time can be obtained from the difference value between the two voltage values in the same phase. This makes it possible to obtain an offset that changes significantly over time.
  • the amount of change in the voltage value per unit time can be obtained from the difference value of the two voltage values in the same phase. This makes it possible to obtain an offset that changes significantly over time.
  • FIG. 3 is a cross-sectional view showing a part of the electrode unit 3 at a position along the IV-IV line of FIG. It is sectional drawing which shows a part of the electrode unit 3 at the position along the VV line of FIG. It is a figure which shows typically the electrical connection in the electric resistance measuring apparatus 1 of embodiment. It is a flow chart of the electric resistance measurement using the electric resistance measuring apparatus 1. It is a figure which shows the waveform 91 (rectangular wave) of the voltage value obtained by the measurement. It is a figure which shows the waveform 93 (sine wave) of the voltage value obtained by the measurement.
  • the plane parallel to the substrate of the electrode holding member is referred to as a horizontal plane
  • the direction parallel to the horizontal plane is referred to as “horizontal direction”
  • the direction perpendicular to the horizontal plane is referred to as “vertical direction”.
  • FIG. 1 is a diagram schematically showing the electrical resistance measuring device 1 and the cell culture vessel 2 of the embodiment.
  • FIG. 2 is a top view of the cell culture vessel 2.
  • FIG. 3 is a top view of the electrode unit 3 of the embodiment.
  • FIG. 4 is a cross-sectional view showing a part of the electrode unit 3 at a position along the IV-IV line of FIG.
  • FIG. 5 is a cross-sectional view showing a part of the electrode unit 3 at a position along the VV line of FIG.
  • the electrical resistance measuring device 1 is a device that measures the electrical resistance (impedance) of cells cultured in the cell culture vessel 2.
  • the electric resistance measuring device 1 includes an electrode unit 3 and a measuring device 4 connected to the electrode unit 3.
  • the electrode unit 3 is placed on the cell culture vessel 2.
  • the cell culture container 2 is a container used for culturing cells.
  • the cell culture vessel 2 is a so-called well plate having a plurality of wells 21.
  • the well 21 is a recess recessed downward from the upper surface of the cell culture vessel 2.
  • a total of 6 wells 21 in 2 rows vertically and 3 columns horizontally are arranged in a grid pattern.
  • the number of wells 21 in the cell culture vessel 2 is not limited to 6, but may be 1 to 5, or 7 or more.
  • An insert cup 23 is installed in the cell culture container 2.
  • one insert cup 23 is arranged inside each of the six wells 21.
  • the insert cup 23 has a tubular portion 25, a support portion 27, and a cell culture portion 29.
  • the tubular portion 25 is an insulating portion formed in a truncated cone shape and a tubular shape.
  • the support portion 27 extends outward from the upper end of the tubular portion 25. When the support portion 27 is placed on the upper surface of the cell culture container 2, the tubular portion 25 and the cell culture portion 29 are arranged inside the well 21 at a position where the cell culture portion 29 does not contact the bottom surface of the well 21. Can be done.
  • the support portions 27 are radially arranged at three locations in the circumferential direction of the tubular portion 25.
  • the support portion 27 may have another shape as long as it can support the tubular portion 25 and the cell culture portion 29 at predetermined positions.
  • the support portions 27 may be arranged at only two locations in the circumferential direction of the insert cup 23, or may be arranged at four or more locations.
  • the support portion 27 may have a flange shape extending outward from a part of the insert cup 23 in the circumferential direction.
  • the cell culture section 29 is a membrane that covers the lower opening of the tube section 25.
  • a membrane having cell adhesion is used for the cell culture unit 29.
  • a plate-shaped member provided with a large number of fine through holes may be used for the cell culture unit 29.
  • the electric resistance measuring device 1 includes an electrode unit 3 and a measuring device 4.
  • the electrode unit 3 has a substrate 31, six first pillars 35, six second pillars 37, and six electrode sets.
  • the electrode set includes working electrodes 41 and 43 and reference electrodes 51 and 53 as a set.
  • the electrode unit 3 is a member for arranging one electrode set in each of the six wells 21 of the cell culture vessel 2.
  • the substrate 31 is a member having a flat plate shape extending in the horizontal direction and having an insulating property.
  • the substrate 31 is made of, for example, a glass epoxy resin.
  • the substrate 31 has a plurality of observation openings 311 arranged in a grid pattern. Each observation opening 311 is provided at a position overlapping each well 21 of the cell culture vessel 2 in the vertical direction.
  • a plurality of lead wires 33 are arranged on the substrate 31.
  • One end of each lead wire 33 is an electrode connection terminal 331.
  • Each lead wire 33 is connected to one of the working electrodes 41 and 43 provided on the first pillar portion 35 and the reference electrodes 51 and 53 provided on the second pillar portion 37 via the electrode connection terminal 331.
  • the other end of each lead wire 33 is an external connection terminal 333.
  • Each lead wire 33 is connected to the measuring instrument 4 via the external connection terminal 333.
  • At least a part of the lead wire 33 including the electrode connection terminal 331 is printed on the upper surface of the substrate 31.
  • the other portion of the lead wire 33 may be printed on the inner layer of the substrate 31 formed in a plurality of layers, or may be embedded in the inner layer at the time of manufacturing the substrate 31.
  • the first and second pillar portions 35 and 37 are provided in the vicinity of each observation opening 311. In this embodiment, one first and one second pillar portion 35, 37 are provided for each of the six observation openings 311. The first and second pillar portions 35, 37 may be provided only for a part of the six observation openings 311.
  • the first and second pillar portions 35 and 37 have a square columnar shape extending in the vertical direction.
  • the end portion of the first pillar portion 35 and the end portion of the second pillar portion 37 are connected to the substrate 31, and the first pillar portion 35 and the second pillar portion 37 extend downward from the portion connected to the substrate 31. ..
  • the first and second pillar portions 35, 37 are members having an insulating property, and are made of, for example, glass.
  • the first and second pillar portions 35, 37 may be made of a glass epoxy resin, similarly to the substrate 31.
  • a working electrode 41 (first working electrode) and a reference electrode 51 (first reference electrode) are provided on the outer surface of the first pillar portion 35. As shown in FIG. 4, of the four side surfaces of the first pillar portion 35, the working electrode 41 is provided on one first side surface 351 and the reference electrode 51 is provided on the second side surface 353 facing the first side surface 351. Provided. The working electrode 41 and the reference electrode 51 are not adjacent to each other and are arranged at intervals.
  • a working electrode 43 (second working electrode) and a reference electrode 53 (second reference electrode) are provided on the outer surface of the second pillar 37. As shown in FIG. 5, of the four side surfaces of the second pillar portion 37, the working electrode 43 is provided on one first side surface 371, and the reference electrode 53 is provided on the second side surface 373 facing the first side surface 371. Provided. The working electrode 43 and the reference electrode 53 are not adjacent to each other and are arranged at intervals.
  • the working electrodes 41, 43 and the reference electrodes 51, 53 are conductive members, preferably metals such as silver-silver chloride, gold (Au), platinum (Pt), or indium tin oxide (ITO). It is composed.
  • the working electrodes 41 and 43 and the reference electrodes 51 and 53 are preferably metal thin films.
  • the metal thin film can be formed, for example, by print formation or vapor deposition.
  • any one or all of the working electrodes 41 and 43 and the reference electrodes 51 and 53 may be used as a plate-shaped or linear conductive member, and the first pillar portion 35 or the second pillar may be formed via an adhesive or the like. It may be attached to the portion 37.
  • the substrate 31 is provided with a plurality of through holes 313.
  • the through hole 313 is provided around the observation opening 311.
  • the first pillar portion 35 and the second pillar portion 37 are inserted into each through hole 313.
  • the upper end portions of the first pillar portion 35 and the second pillar portion 37 project upward from the upper surface of the substrate 31.
  • the electrode connection terminals 331 of the two lead wires 33 are arranged at positions adjacent to the through holes 313. As shown in FIG. 4, the working electrode 41 and the reference electrode 51 are each electrically connected to the electrode connection terminal 331 of the conducting wire 33 via the conductive connecting member 315.
  • Each conductive connecting member 315 has a substantially rectangular parallelepiped shape. As shown in the enlarged portions, FIGS. 4 and 5, in FIG. 3, the conductive connecting member 315 is arranged on the electrode connecting terminal 331 of each conducting wire 33. Further, as shown in FIG. 4, the vicinity of the upper end portion of the working electrode 41 is adhered to the side surface of one conductive connecting member 315 by the conductive adhesive 317. Further, the lower surface of the conductive connecting member 315 and the upper surface of the electrode connecting terminal 331 of one conducting wire 33 are adhered to each other by the conductive adhesive 317. As a result, the working electrode 41 and one conducting wire 33 are electrically connected via the conductive connecting member 315 and the conductive adhesive 317.
  • the vicinity of the upper end of the reference electrode 51 is adhered to the side surface of the other conductive connecting member 315 by the conductive adhesive 317. Further, the lower surface of the conductive connecting member 315 and the upper surface of the electrode connecting terminal 331 of the other conducting wire 33 are adhered to each other by the conductive adhesive 317. As a result, the reference electrode 51 and the other conductor 33 are electrically connected via the conductive connecting member 315 and the conductive adhesive 317.
  • the working electrode 41 and the reference electrode 51 are adhered to the substrate 31 via the conductive connecting member 315, so that the upper end portion of the first pillar portion 35 is fixed to the substrate 31.
  • the working electrode 43 and the reference electrode 53 are also similarly connected to the conductor 33 via the conductive connecting member 315.
  • the insert cup 23 is moved to one side in the horizontal direction (right side in FIGS. 1 to 3) in each well 21. Be placed. Then, by placing the electrode unit 3 on the cell culture vessel 2, the lower end portion of the first pillar portion 35 and the lower end portion of the second pillar portion 37 are arranged inside the well 21.
  • each observation opening 311 is arranged substantially in the center of each well 21.
  • the first pillar portion 35 is arranged on one side of the observation opening 311 (on the right side in FIG. 3), and the second pillar portion 37 is on the other side of the observation opening 311 (in FIG. 3). Then it is placed on the left side). Since the insert cup 23 is unevenly arranged on one side of the well 21, the first pillar portion 35 is inserted inside the insert cup 23, and the second pillar portion 37 is inserted outside the insert cup 23. ..
  • the second pillar portion 37 projects downward from the first pillar portion 35.
  • the lower end of the first column 35 is arranged above the cell culture 29 of the insert cup 23, and the lower end of the second column 37 is the cell culture. It is arranged below the portion 29. That is, the lower ends of the working electrode 41 and the reference electrode 51 are arranged above the cell culture section 29 of the well 21, and the lower ends of the working electrode 43 and the reference electrode 53 are located below the cell culture section 29. Will be done.
  • FIG. 6 is a diagram schematically showing an electrical connection in the electrical resistance measuring device 1 of the embodiment.
  • the measuring instrument 4 includes an AC power supply 61, a voltmeter 63, and a control unit 7.
  • the AC power supply 61 is connected to the working electrodes 41 and 43 via the lead wire 33, and applies an AC signal having a fixed cycle.
  • the voltmeter 63 is connected to the reference electrodes 51 and 53 via the lead wire 33, and measures the potential difference between the reference electrodes 51 and 53.
  • the measuring instrument 4 may include a switching circuit (not shown) for switching the electrode set connected to the AC power supply 61 and the voltmeter 63 among the six electrode sets. The operation of the switching circuit may be controlled by the control unit 7.
  • FIG. 6 shows a state in which one of the six electrode sets is connected to the AC power supply 61 and the voltmeter 63.
  • the control unit 7 is configured as a general computer equipped with a CPU, RAM, and an auxiliary storage device (for example, a hard disk) (not shown).
  • the CPU executes various processes for measuring electrical resistance by operating according to a program installed in the auxiliary storage device.
  • a dedicated circuit such as an integrated circuit (ASIC) for a specific application, various functions of the control unit 7 may be realized in terms of hardware.
  • ASIC integrated circuit
  • the control unit 7 is electrically controlled with the AC power supply 61 and controls the operation of the AC power supply 61.
  • the control unit 7 is electrically connected to the voltmeter 63, and receives an input signal representing a potential difference (voltage value) from the voltmeter 63.
  • the electrical resistance calculation unit 71 shown in FIG. 6 is a function realized by operating the CPU according to a program.
  • the electric resistance calculation unit 71 has an offset calculation unit 73.
  • the offset calculation unit 73 performs a process of calculating an offset included in the voltage value obtained by the voltmeter 63. The process executed by the electric resistance calculation unit 71 will be described later.
  • the electric resistance calculation unit 71 performs a correction process for removing the offset from the voltage value obtained by the voltmeter 63, and also executes a process for obtaining the electric resistance of the cell layer 9 from the corrected voltage value.
  • the control unit 7 including the electric resistance calculation unit 71 is an example of the electric resistance calculation device.
  • the insert cup 23 in which the cell layer 9 to be measured is formed on the cell culture unit 29 is used for cell culture. It is set in the well 21 of the container 2. Then, the electrode unit 3 is placed on the cell culture container 2. Each well 21 is provided with a conductive solution (culture solution, etc.) to the extent that the tips of the working electrodes 41, 43 and the reference electrodes 51, 53 provided on the first pillar 35 and the second pillar 37 are immersed. Infused. In this state, the measuring instrument 4 is connected to each lead wire 33 of the electrode unit 3. As a result, the circuit shown in FIG. 6 is formed, and the electrical resistance of the cell layer 9 can be measured.
  • a conductive solution culture solution, etc.
  • the electrode unit 3 is provided with an observation opening 311 corresponding to each well 21. This makes it easy to observe the cells cultured on the cell culture unit 29. Further, by arranging the first and second pillars 35 and 37 at positions that do not overlap the observation opening 311 in the vertical direction, the measurement of the electrical resistance of the cell layer 9 and the observation of the cell layer 9 can be performed at the same time. Can be done.
  • the resistance Rm is the electricity of the cell culture unit 29 and the cell layer 9 cultured on the cell culture unit 29 (hereinafter, the cell culture unit 29 and the cell layer 9 are collectively referred to as a “cell unit”).
  • the resistance Rw1 corresponds to the electrical resistance of the culture solution between the working electrode 41 and the cell portion.
  • the resistance Rw2 corresponds to the electrical resistance of the culture medium between the working electrode 43 and the cell portion.
  • the resistance Rr1 corresponds to the electrical resistance of the culture medium between the reference electrode 51 and the cell portion.
  • the resistance Rr2 corresponds to the electrical resistance of the culture medium between the reference electrode 53 and the cell portion.
  • the electrical resistance values of the resistances Rw1, Rr1, Rw2, Rr2 and the resistance Rm of only the cell culture unit 29 in the absence of the cell layer 9 are measured in advance as controls. Will be done.
  • FIG. 7 is a flow chart of electrical resistance measurement using the electrical resistance measuring device 1. Unless otherwise specified, each process shown in FIG. 7 shall be executed under the control of the control unit 7. In the electric resistance measurement, the potential difference measurement step S1, the difference value calculation step S2, the change amount calculation step S3, the correction step S4, and the electric resistance calculation step S5 are executed in this order.
  • the potentiometric titration measurement step S1 is executed in a state where the electrical resistance of the cell layer 9 can be measured.
  • the control unit 7 drives the AC power supply 61 to apply an AC signal having a constant cycle T between the working electrodes 41 and 43. Further, the control unit 7 acquires the voltage value measured by the voltmeter 63.
  • the voltage value for at least one cycle is measured by applying the AC signal for one cycle or more continuously in time.
  • FIG. 8 is a diagram showing a waveform 91 (rectangular wave) of a voltage value obtained by measurement.
  • the horizontal axis represents time (phase) and the vertical axis represents voltage.
  • the voltage value measured by the voltmeter 63 becomes a square wave waveform 91.
  • a linear increase (offset) of the voltage value occurs in the waveform 91. This offset increases linearly over time. This offset is considered to be due to an increase in the apparent voltage value due to the accumulation of charges in the electric double layer at the reference electrodes 51 and 53. It is difficult to accurately determine the electrical resistance of the cell layer 9 with the voltage value including this offset. Further, since the application of the AC signal may damage the cell layer 9, it is desirable to obtain the electric resistance from the voltage value obtained as soon as possible after the application of the AC signal.
  • the electrical resistance measuring device 1 executes a process of removing the offset from the measured voltage value. Specifically, first, the difference value calculation step S2 shown in FIG. 7 is performed. In the difference value calculation step S2, the offset calculation unit 73 executes a process of calculating the difference value of two voltage values in the same phase from the voltage value obtained in the potential difference measurement step S1.
  • the waveform 91 shown in FIG. 8 is obtained by the potentiometric titration measurement step S1.
  • the waveform 91 is a waveform for two cycles from time 0 to time 2T.
  • the time 0 to the time T is referred to as the first cycle T1
  • the time T to the time 2T is referred to as the second cycle T2.
  • the offset calculation unit 73 selects, for example, the voltage value V1 (point P1) of the time T / 4 in the first period T1 as the first voltage value.
  • two voltage values V1 and V2 having a time difference T for one cycle are selected as voltage values having the same phase, but the time difference is two or more cycles.
  • Two voltage values may be selected. That is, in the difference value calculation step S2, the difference value between the two voltage values, which is the time difference for n cycles, is obtained, where n is a natural number of 1 or more. Then, in the change amount calculation step S3, the difference value obtained in the difference value calculation step S2 is divided by the time difference between the measurement times of the two voltage values (that is, for n cycles) to obtain the offset change amount per unit time. K is required.
  • the offset change amount K may be obtained for each different phase, and the representative value (mean value, median value, etc.) of the obtained plurality of offset change amounts K may be used as the final offset change amount K.
  • the offset change amount K in the phase corresponding to T / 4 is obtained, but the phase is different from T / 4 (for example, T / 2 or 3T / 4).
  • the offset change amount K at the corresponding phase) may also be obtained.
  • the representative value of the obtained plurality of offset change amounts K may be used as the final offset change amount K.
  • the offset calculation unit 73 executes a process for obtaining the offset, and the electric resistance calculation unit 71 performs a correction process for removing the obtained offset from the voltage value obtained by the voltmeter 63.
  • the electric resistance calculation unit 71 subtracts the offset obtained by the linear equation f (t) from the voltage value at each time t obtained in the potentiometric titration measurement step S1. As a result, a voltage value corrected for the offset is obtained.
  • the electric resistance calculation unit 71 calculates the electric resistance by dividing the corrected voltage value by the current value of the AC signal.
  • the waveform 91r shown in FIG. 8 is a waveform obtained by a correction process excluding the offset in the waveform 91. In the waveform 91r, the offset component that increases linearly with time is removed. Therefore, by calculating the electric resistance from the voltage value indicated by the waveform 91r, the electric resistance of the cell layer 9 can be obtained with high accuracy.
  • the measurement times of the two voltage values selected in the difference value calculation step S2 are set to the first time t1 and the second time t2, respectively. Let t1 ⁇ t2). Then, according to f (t) based on the offset change amount K, the offset during the period from the first time t1 to the second time t2 can be accurately obtained, so that the voltage value during the period is accurately corrected. It is thought that it can be done. Therefore, in the electric resistance calculation step S5, it is desirable that the electric resistance calculation unit 71 obtains the electric resistance from the voltage value within the period from the first time t1 to the second time t2. For example, in the example shown in FIG.
  • the electric resistance calculation unit 71 calculates the electric resistance from the voltage value within the period from T / 4 to 5T / 4 in the corrected waveform 91r, thereby accurately determining the electric resistance of the cell layer 9. It is thought that it can be sought. However, it is not prevented that the electric resistance calculation unit 71 obtains the electric resistance from the voltage value measured in the period other than the period from the time T / 4 to the time 5 T / 4.
  • FIG. 9 is a diagram showing a waveform 93 (sine wave) of a voltage value obtained by measurement.
  • the horizontal axis represents time (phase) and the vertical axis represents voltage.
  • the measured voltage value becomes a sinusoidal waveform 93.
  • the offset of the voltage value generated at the time of measuring the electric resistance can be appropriately corrected, so that the electric resistance of the cell layer 9 can be obtained with high accuracy.
  • the electrode When the electrode is composed of silver-silver chloride, it is necessary to first prepare the silver electrode and then carry out the chlorination treatment on the silver electrode. Therefore, the manufacturing cost of the electrode increases.
  • the electrode when the electrode is made of gold, platinum, or indium tin oxide, an additional treatment corresponding to the chlorination treatment is not particularly required, so that the manufacturing cost of the electrode can be suppressed.
  • gold, platinum, or indium tin oxide metal electrodes are used for the reference electrodes 51 and 53, the influence of the electric double layer capacitance becomes larger than when silver-silver chloride is used.
  • the electric resistance measuring device 1 the offset can be appropriately corrected, so that even when a metal electrode of gold, platinum, or indium tin oxide is adopted, the electricity of the cell layer 9 is obtained. The resistance can be calculated properly.
  • the target for measuring the electrical resistance by the electrical resistance measuring device 1 is the cell layer 9, but a living tissue may be used.
  • each lead wire 33 is not limited to the one printed on the substrate 31. Further, the arrangement of the lead wire 33 shown in FIG. 3 can be changed as appropriate.
  • the detailed configuration of the electrode unit 3 may differ from each drawing of the present application. Further, the elements appearing in the above-described embodiments and modifications may be appropriately combined as long as there is no contradiction.

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Abstract

La présente invention concerne un dispositif de mesure de résistance électrique (1) comprenant des électrodes de travail (41, 43), des électrodes de référence (51, 53), une alimentation électrique CA (61) qui est connectée aux électrodes de travail (41, 43) et applique un signal CA ayant une fréquence fixe, un voltmètre (63) qui est connecté aux électrodes de référence (51, 53) et mesure une différence de potentiel, et une unité de calcul de résistance électrique (71) qui calcule la résistance électrique d'une couche de cellules (9) sur la base d'une valeur de tension obtenue par le voltmètre (63). L'unité de calcul de résistance électrique (71) comprend une unité de calcul de décalage (73) qui exécute un processus pour calculer une quantité de changement de la valeur de tension par unité de temps à partir d'une valeur de différence entre deux valeurs de tension dans la même phase.
PCT/JP2020/032922 2019-09-20 2020-08-31 Dispositif de mesure de résistance électrique, procédé de mesure de résistance électrique, et dispositif de calcul de résistance électrique WO2021054103A1 (fr)

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JP2003531373A (ja) * 2000-04-14 2003-10-21 ナノテック ソリューション バイオマスの特性を決定する装置および方法
US20050221274A1 (en) * 2004-03-31 2005-10-06 Negulescu Paul A Multiwell plate assembly for use in high throughput assays
JP2011115097A (ja) * 2009-12-03 2011-06-16 Hokkaido Univ 高分子化合物またはその複合体の細胞透過性評価装置およびその細胞透過性評価方法
US20120267260A1 (en) * 2011-04-25 2012-10-25 Sameera Dharia Monitoring membrane-bound proteins
WO2012147463A1 (fr) * 2011-04-28 2012-11-01 株式会社日立製作所 Récipient pour culture cellulaire et appareil associé
US20150068926A1 (en) * 2012-04-13 2015-03-12 Smartcare Technologies Ltd Electrical impedance hematocrit and hba1c biosensor comprising sample plate and sample apparatus

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Publication number Priority date Publication date Assignee Title
JP2003531373A (ja) * 2000-04-14 2003-10-21 ナノテック ソリューション バイオマスの特性を決定する装置および方法
US20050221274A1 (en) * 2004-03-31 2005-10-06 Negulescu Paul A Multiwell plate assembly for use in high throughput assays
JP2011115097A (ja) * 2009-12-03 2011-06-16 Hokkaido Univ 高分子化合物またはその複合体の細胞透過性評価装置およびその細胞透過性評価方法
US20120267260A1 (en) * 2011-04-25 2012-10-25 Sameera Dharia Monitoring membrane-bound proteins
WO2012147463A1 (fr) * 2011-04-28 2012-11-01 株式会社日立製作所 Récipient pour culture cellulaire et appareil associé
US20150068926A1 (en) * 2012-04-13 2015-03-12 Smartcare Technologies Ltd Electrical impedance hematocrit and hba1c biosensor comprising sample plate and sample apparatus

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