WO2020129452A1 - Dispositif et procédé de mesure - Google Patents

Dispositif et procédé de mesure Download PDF

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Publication number
WO2020129452A1
WO2020129452A1 PCT/JP2019/043730 JP2019043730W WO2020129452A1 WO 2020129452 A1 WO2020129452 A1 WO 2020129452A1 JP 2019043730 W JP2019043730 W JP 2019043730W WO 2020129452 A1 WO2020129452 A1 WO 2020129452A1
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WO
WIPO (PCT)
Prior art keywords
working
current
group
working electrode
electrode
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PCT/JP2019/043730
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English (en)
Japanese (ja)
Inventor
宏太 宮川
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株式会社ヨコオ
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Priority to JP2020561209A priority Critical patent/JPWO2020129452A1/ja
Publication of WO2020129452A1 publication Critical patent/WO2020129452A1/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
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems

Definitions

  • the present invention relates to a measuring device and a measuring method for electrochemical measurement, and particularly to a measuring device and a measuring method for measuring a redox current.
  • Patent Document 1 describes an example of a measuring device using electrochemical measurement.
  • This measuring device includes a single reference electrode, a single counter electrode, and a plurality of working electrodes.
  • the reference electrode and the counter electrode are electrically connected to the potentiostat.
  • the plurality of working electrodes are electrically connected to the multiplexer.
  • the multiplexer sequentially switches the working electrode electrically connected to the potentiostat from the plurality of working electrodes.
  • One example of the object of the present invention is to reduce the number of current-voltage conversion circuits while reducing the number of times of switching the electrical connection of the working electrode-current-voltage conversion circuit.
  • Other objects of the present invention will be apparent from the description herein.
  • a measuring device for measuring a redox current A first switching circuit, A second switching circuit, A first group of working electrodes electrically connected to the first switching circuit; A second group of working electrodes electrically connected to the second switching circuit; Equipped with The first switching circuit sequentially switches the working electrodes electrically connected to the first current-voltage conversion circuit from the working electrodes of the first group, The second switching circuit sequentially switches the working electrodes electrically connected to the second current-voltage conversion circuit from the working electrodes of the second group,
  • the first group of working electrodes and the second group of working electrodes are a measuring device arranged in a common well.
  • Another aspect of the present invention is A measuring method for measuring a redox current,
  • the working electrodes electrically connected to the first current-voltage conversion circuit among the working electrodes of the first group located in the well and electrically connected to the first switching circuit are sequentially switched by the first switching circuit
  • the working electrodes electrically connected to the second current-voltage conversion circuit among the working electrodes of the second group located in the well and electrically connected to the second switching circuit are sequentially switched by the second switching circuit.
  • FIG. 3 is a sectional view taken along the line AA′ of FIG. 2. It is a figure for demonstrating the measuring method by a measuring device. It is a figure for demonstrating the detail of the block shown in FIG.
  • FIG. 6 is a sectional view taken along the line PP′ of FIG. 5. It is a figure for demonstrating the connection between a working electrode and a counter electrode in a well.
  • FIG. 1 is a circuit diagram of a measuring device 20 according to the embodiment.
  • FIG. 2 is a plan view of the element 10 according to the embodiment.
  • FIG. 3 is a sectional view taken along the line AA′ of FIG.
  • FIG. 4 is a diagram for explaining a measuring method by the measuring device 20.
  • the measuring device 20 measures a redox current.
  • the measuring device 20 includes a plurality of working electrode groups G (first working electrode Ga and second working electrode Gb) and a plurality of switching circuits 400 (first switching circuit 400a and second switching circuit 400b).
  • the first group of working electrodes Ga and the second group of working electrodes Gb are located in a common well (well 300) and are electrically connected to the first switching circuit 400a and the second switching circuit 400b, respectively.
  • the first switching circuit 400a electrically connects the working electrode Ga of the first group to the first current-voltage converting circuit 510a (one of the plurality of current-voltage converting circuits 510). 112 are sequentially switched.
  • the second switching circuit 400b is electrically connected to the second current-voltage conversion circuit 510b (another current-voltage conversion circuit of the plurality of current-voltage conversion circuits 510) from the second group of working electrodes Gb.
  • the working electrode 112 is sequentially switched.
  • the number of current-voltage conversion circuits 510 while reducing the number of times of switching the electrical connection between the working electrode 112 and the current-voltage conversion circuit 510.
  • the number of times the electrical connection between the working electrode 112 and the current-voltage conversion circuit 510 is switched is such that the working electrode Ga of the first group and the working electrode Gb of the second group are common. The number can be reduced as compared with the case where 400 is connected.
  • the number of current-voltage conversion circuits 510 is the same as the number of current-voltage conversion circuits 510 for each of the plurality of working electrodes 112 in the first group working electrode Ga and the second group working electrode Gb. It is possible to reduce the number as compared with the case where the switching circuit 400 is not provided, that is, the case where the switching circuit 400 is not provided. In particular, the greater the number of working electrodes 112 located in the common well (well 300), the more meaningful the above-mentioned configuration is. The number of working electrodes 112 located in the common well (well 300) may be four or more, for example.
  • the first switching circuit 400a may electrically connect the first working electrode 112a in the working electrode Ga of the first group to the first current-voltage conversion circuit 510a at the first timing, and after the first timing, At two timings, the second working electrode 112b in the first group of working electrodes Ga may be electrically connected to the first current-voltage conversion circuit 510a.
  • the second switching circuit 400b may electrically connect the third working electrode 112c in the second group of working electrodes Gb to the second current-voltage conversion circuit 510b at substantially the same timing as the first timing.
  • the fourth working electrode 112d in the second group of working electrodes Gb may be electrically connected to the second current-voltage conversion circuit 510b at substantially the same timing as the second timing.
  • the redox current of the first working electrode 112a and the redox current of the third working electrode 112c can be measured at substantially the same timing, and the redox current of the second working electrode 112b can be measured.
  • the redox current and the redox current of the fourth working electrode 112d can be measured at substantially the same timing.
  • the timing at which the third working electrode 112c is electrically connected to the second current-voltage conversion circuit 510b may not be exactly the same as the timing at which the first working electrode 112a is electrically connected to the first current-voltage conversion circuit 510a. Good.
  • the voltage applied by the controller 610 (which will be described later with reference to FIG. 1) from the timing when the first working electrode 112a is electrically connected to the first current-voltage conversion circuit 510a.
  • the third working electrode 112c may be electrically connected to the second current-voltage conversion circuit 510b before the change. The same applies to the timing when the fourth working electrode 112d is electrically connected to the second current-voltage conversion circuit 510b and the timing when the second working electrode 112b is electrically connected to the second current-voltage conversion circuit 510b.
  • the first working electrode 112a, the second working electrode 112b, the third working electrode 112c, and the fourth working electrode 112d are respectively a first distance D1, a second distance D2, a third distance D3, and a fourth distance from the counter electrode 212. D4 away.
  • the third distance D3 may be equal to or close to the first distance D1, and may be, for example, 80% or more and 120% or less, preferably 90% or more and 110% or less of the first distance D1.
  • the fourth distance D4 may be equal to or close to the second distance D2, and may be, for example, 80% or more and 120% or less, preferably 90% or more and 110% or less of the second distance D2.
  • the solution resistance in the well 300 depends on the distance between the working electrode 112 and the counter electrode 212.
  • the first working electrode 112a and the third working electrode 112c which are measured at substantially the same timing, are in the range of substantially the same distance from the counter electrode 212. Therefore, it is possible to suppress an error caused by the solution resistance between the measurement results of the first working electrode 112a and the third working electrode 112c. The same applies to the second working electrode 112b and the fourth working electrode 112d.
  • the counter electrode 212 may be located at the center of the first surface 102 of the substrate 100, as described later with reference to FIG.
  • the first working electrode 112a, the second working electrode 112b, the third working electrode 112c, and the fourth working electrode 112d are respectively a first distance D1 and a second distance D2 from the center of the first surface 102 of the substrate 100.
  • the measuring device 20 includes a plurality of working electrodes 112, a counter electrode 212, a reference electrode 222, a plurality of switching circuits 400, a potentiostat 500, a controller 610, and a plurality of measuring devices 620.
  • the redox current is measured by the plurality of working electrodes 112, the counter electrode 212, and the reference electrode 222.
  • the plurality of working electrodes 112, the counter electrode 212, and the reference electrode 222 are located in a common well (well 300) and constitute an electrochemical cell.
  • the measuring device 20 includes a plurality of working electrode groups G.
  • Each of the plurality of working electrode groups G includes a plurality of working electrodes 112.
  • Each of the plurality of working electrode groups G is electrically connected to each of the plurality of switching circuits 400.
  • the counter electrode 212, the reference electrode 222, and the plurality of switching circuits 400 are electrically connected to the potentiostat 500.
  • the potentiostat 500 includes a plurality of current-voltage conversion circuits 510, an operational amplifier 522, an operational amplifier 524, a resistor 526, and a resistor 528.
  • Each of the plurality of switching circuits 400 is electrically connected to each of the plurality of current-voltage conversion circuits 510.
  • Each of the plurality of switching circuits 400 sequentially switches the working electrode 112 electrically connected to the corresponding current-voltage conversion circuit 510 from the corresponding working electrode group G.
  • the current-voltage conversion circuit 510 converts a current (oxidation-reduction current) flowing from the working electrode 112 via the switching circuit 400 into a voltage.
  • the switching circuit 400 is a semiconductor switch, more specifically, an analog multiplexer.
  • the switching by the switching circuit 400 may be performed by, for example, a microcomputer mounted on the measuring device 20, or by a computer provided outside the measuring device 20 (for example, a desktop computer or a laptop computer). Good.
  • the switching circuit 400 is not limited to the example shown in FIG. 1 and may be, for example, a relay circuit or a mechanical switch.
  • the current-voltage conversion circuit 510 includes an operational amplifier 512 and a resistor 514.
  • the resistor 514 is electrically connected between the output terminal and the inverting input terminal of the operational amplifier 512, and functions as a feedback resistor.
  • the non-inverting input terminal of the operational amplifier 512 is grounded.
  • the circuit structure of the current-voltage conversion circuit 510 is not limited to the example shown in FIG.
  • Each of the plurality of current-voltage conversion circuits 510 is connected to each of the plurality of measuring instruments 620.
  • the measuring device 620 measures the voltage output from the output terminal of the corresponding current-voltage conversion circuit 510.
  • the measuring device 620 is, for example, a voltmeter.
  • the counter electrode 212 is electrically connected to the output terminal of the operational amplifier 522.
  • the non-inverting input terminal of the operational amplifier 522 is grounded.
  • a voltage is input to the inverting input terminal of the operational amplifier 522 from the controller 610 (for example, a function generator) via the resistor 526.
  • the reference electrode 222 is connected to the inverting input terminal of the operational amplifier 522 via the operational amplifier 524 and the resistor 528.
  • the operational amplifier 524 functions as a voltage follower, the non-inverting input terminal of the operational amplifier 524 is electrically connected to the reference electrode 222, and the inverting input terminal of the operational amplifier 524 is electrically connected to the output terminal of the operational amplifier 524. It is connected.
  • the circuit structure of the potentiostat 500 is not limited to the example shown in FIG.
  • the element 10 is a chip.
  • the element 10 includes a substrate 100 and a wiring layer 120 (resist 122).
  • the substrate 100 has a first surface 102 and a second surface 104.
  • the wiring layer 120 is located on the first surface 102 of the substrate 100.
  • the conductive layer 110 (working electrode 112, terminal 114, and wiring 116) is located in the wiring layer 120.
  • the second surface 104 is on the opposite side of the first surface 102.
  • the well 300 is defined by the partition wall 302 on the first surface 102 side of the substrate 100.
  • a sample (solution) can be placed in the well 300.
  • the counter electrode 212 is held above the element 10. In this way, the counter electrode 212 can contact the sample in the well 300. The same applies to the reference electrode 222.
  • the substrate 100 (first surface 102) has a substantially quadrangular shape.
  • This quadrangle can be, for example, a rectangle.
  • the substrate 100 has a first side 106a, a second side 106b, a third side 106c, and a fourth side 106d.
  • the second side 106b is on the opposite side of the first side 106a.
  • the third side 106c is between the first side 106a and the second side 106b.
  • the fourth side 106d is on the opposite side of the third side 106c.
  • the quadrangle of the substrate 100 does not have to be a strict quadrangle, and may have, for example, sides with cuts or have rounded corners.
  • the substrate 100 may have a shape other than a quadrangle.
  • the element 10 includes a plurality of blocks B.
  • the element 10 includes eight blocks B.
  • the two blocks B are arranged along the first side 106a
  • the two blocks B are arranged along the second side 106b
  • the two blocks B are arranged along the third side 106c.
  • the two blocks B are arranged along the fourth side 106d.
  • the layout of the plurality of blocks B is not limited to the example shown in FIG.
  • Each of the plurality of blocks B includes a plurality of conductive layers 110.
  • each of the plurality of blocks B includes 15 conductive layers 110.
  • Each of the plurality of conductive layers 110 has a working electrode 112, a terminal 114, and a wiring 116.
  • the plurality of working electrodes 112 overlap the well 300.
  • the plurality of terminals 114 do not overlap the well 300 and are located outside the well 300.
  • Each of the plurality of wirings 116 connects each of the plurality of working electrodes 112 to each of the plurality of terminals 114.
  • the plurality of terminals 114 can be electrically connected to the switching circuit 400 (FIG. 1).
  • the switching circuit 400 may be arranged outside the substrate 100, for example.
  • the working electrode 112 can be electrically connected to the switching circuit 400 via the wiring 116 and the terminal 114.
  • the layout of the conductive layer 110 is not limited to the example shown in FIG.
  • the plurality of working electrodes 112 are arranged in a matrix of m rows and n columns (each of m and n is independently an integer). In the example shown in FIG. 2, 15 working electrodes 112 are arranged in a matrix of 3 rows and 5 columns. Further, the plurality of working electrodes 112 in each block B are arranged in a substantially same rule among the plurality of blocks B (for example, the number of rows and columns of the matrix or the spacing between the adjacent working electrodes 112). .. However, the layout of the plurality of working electrodes 112 is not limited to the example shown in FIG.
  • each of the plurality of blocks B includes two working electrode groups G.
  • One working electrode group G includes eight working electrodes 112, and another working electrode group G includes seven working electrodes 112.
  • One working electrode group G is electrically connected to one switching circuit 400 (FIG. 1), and the other one working electrode group G is electrically connected to another one switching circuit 400 (FIG. 1). It is connected to the.
  • the number of working electrode groups G in each block B is not limited to the example shown in FIG.
  • each block B may include only one working electrode group G, or may include three or more working electrode groups G.
  • the counter electrode 212 is located at the center of the first surface 102 of the substrate 100, and the reference electrode 222 is located off the center of the first surface 102 of the substrate 100.
  • the layout of the counter electrode 212 and the reference electrode 222 is not limited to the example shown in FIG.
  • Each of the plurality of switching circuits 400 sequentially switches the working electrodes 112 electrically connected to the corresponding current-voltage conversion circuit 510 from the corresponding working electrode group G at substantially the same timing.
  • the working electrodes 112 numbered 1 to 8 in the working electrode Ga of the first group are sequentially electrically connected to the first current-voltage conversion circuit 510a.
  • the second switching circuit 400b sequentially electrically connects the working electrodes 112 numbered 1 to 8 in the second group of working electrodes Gb to the second current-voltage conversion circuit 510b.
  • the working electrodes 112 numbered 1 to 8 in the working electrode Gb of the second group are respectively the working electrodes 112 numbered 1 to 8 in the working electrode Ga of the first group. Is electrically connected to the current-voltage conversion circuit 510 at substantially the same timing.
  • switching by the switching circuit 400 may be performed in the order of numbers 1 to 8 shown in FIG. 4, or may be performed in an order different from the order of numbers 1 to 8 shown in FIG.
  • the plurality of working electrodes 112 in each working electrode group G are arranged by substantially the same rule among the plurality of working electrode groups G (for example, the number of rows and columns of the matrix or the spacing between adjacent working electrodes 112). You can leave.
  • the working electrodes 112 numbered 1 to 8 in the working electrode Gb of the second group are the working electrodes 112 numbered 1 to 8 in the working electrode Ga of the first group. It is lined up with the substantially same rule.
  • the plurality of switching circuits 400 may electrically connect the working electrodes 112 located at substantially the same position in the working electrode group G to the current-voltage conversion circuit 510 at substantially the same timing.
  • the working electrodes 112 numbered 1 to 8 in the working electrode Gb of the second group are respectively the actions numbered 1 to 8 in the working electrode Ga of the first group.
  • the electrode 112 and the working electrode group G are relatively at substantially the same position in each working electrode group G.
  • FIG. 5 is a diagram for explaining the details of the block B shown in FIG.
  • FIG. 6 is a sectional view taken along line PP′ of FIG.
  • the element 10 includes a substrate 100, a conductive layer 110, and a wiring layer 120 (resist 122).
  • the substrate 100 is, for example, a glass substrate, a semiconductor substrate (for example, a silicon substrate), or a resin substrate.
  • the substrate 100 has a first surface 102 and a second surface 104.
  • the conductive layer 110 and the wiring layer 120 are located on the first surface 102 of the substrate 100.
  • the second surface 104 is on the opposite side of the first surface 102.
  • the conductive layer 110 is made of metal, for example.
  • the conductive layer 110 has a working electrode 112, a terminal 114, and a wiring 116.
  • the working electrode 112 and the terminal 114 are connected to each other via the wiring 116.
  • the structures of the working electrode 112 and the terminal 114 are not limited to the examples shown in FIGS. 5 and 6.
  • the terminal 114 may be located on the second surface 104 side of the substrate 100, and the first surface 102 of the substrate 100 may be provided through a through hole that penetrates the substrate 100 from the first surface 102 to the second surface 104. It may be electrically connected to the working electrode 112 on the side.
  • the resist 122 is made of, for example, an insulating material (for example, resin).
  • the resist 122 has an opening exposing at least a part of the working electrode 112 and an opening exposing at least a part of the terminal 114, and covers at least a part of the wiring 116.
  • the area of the portion of the working electrode 112 exposed from the resist 122 can be reduced, and can be, for example, 200,000 ⁇ m 2 or less.
  • the shape of the portion of the working electrode 112 exposed from the resist 122 may be a circle, and the diameter of the circle may be, for example, 500 ⁇ m or less.
  • FIG. 7 is a diagram for explaining the connection between the working electrode 112 and the counter electrode 212 in the well 300.
  • the connection between the working electrode 112 and the counter electrode 212 can be shown by an equivalent circuit shown in FIG.
  • the equivalent circuit shown in FIG. 7 includes a resistor 312, a resistor 314, and a capacitor 316.
  • the resistor 314 and the capacitor 316 are connected in parallel.
  • the resistor 312 is connected in series with a parallel circuit of the resistor 314 and the capacitor 316.
  • the resistor 312 corresponds to a solution resistance
  • the resistor 314 corresponds to a charge transfer resistor
  • the capacitor 316 corresponds to an electric double layer.
  • the resistance 312 (solution resistance) depends on the distance between the working electrode 112 and the counter electrode 212. Therefore, as described above, when the plurality of working electrodes 112 measured at substantially the same timing are in the range of the substantially same distance from the counter electrode 212, the solution between the measurement results of the plurality of working electrodes 112 is measured. The error due to the resistance can be suppressed.
  • the measuring device 20 can be used for various electrochemical measurements, particularly reversible electrochemical measurements.
  • the measuring device 20 may be used in a device that detects the degree of hybridization of a nucleic acid (for example, micro RNA (miRNA)) based on the measured redox current.
  • a nucleic acid for example, micro RNA (miRNA)
  • the measuring device 20 detects cyclic voltammetry (CV) before nucleic acid hybridization and CV after nucleic acid hybridization.
  • CV cyclic voltammetry

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Abstract

La présente invention concerne un premier groupe d'électrodes de travail (Ga) et un second groupe d'électrodes de travail (Gb) qui sont positionnés dans le même puits (puits (300)) et sont respectivement raccordés électriquement à un premier circuit de commutation (400a) et à un second circuit de commutation (400b). Le premier circuit de commutation (400a) commute séquentiellement l'électrode de travail (112), parmi le premier groupe d'électrodes de travail (Ga), qui est raccordée électriquement à un premier circuit de conversion courant-tension (510a) (un circuit de conversion courant-tension unique parmi une pluralité de circuits de conversion courant-tension (510)). Le second circuit de commutation (400b) commute séquentiellement l'électrode de travail (112), parmi le second groupe d'électrodes de travail (Gb), qui est raccordée électriquement à un second circuit de conversion courant-tension (510b) (un autre circuit de conversion courant-tension unique parmi la pluralité de circuits de conversion courant-tension (510)).
PCT/JP2019/043730 2018-12-18 2019-11-07 Dispositif et procédé de mesure WO2020129452A1 (fr)

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JP2020561209A JPWO2020129452A1 (ja) 2018-12-18 2019-11-07 計測装置及び計測方法

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JP2018-236307 2018-12-18
JP2018236307 2018-12-18

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WO2020129452A1 true WO2020129452A1 (fr) 2020-06-25

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007514175A (ja) * 2003-12-15 2007-05-31 ジーンオーム サイエンシーズ、インク． 多重型電気化学的検出システムと方法
JP2009510446A (ja) * 2005-09-29 2009-03-12 コンビマトリックス・コーポレイション 電極マイクロアレイ上の結合事象の測定方法および装置
JP2010529007A (ja) * 2007-05-25 2010-08-26 ウニベルシダッド アウトノマ デ マドリッド 核酸配列の電気化学的検出法
JP2011099867A (ja) * 2002-07-31 2011-05-19 Toshiba Corp 塩基配列自動解析装置
JP2012047536A (ja) * 2010-08-25 2012-03-08 Nagoya Univ 電流検出装置
WO2017154801A1 (fr) * 2016-03-11 2017-09-14 パナソニックＩｐマネジメント株式会社 Système de mesure électrochimique, dispositif de mesure électrochimique, et procédé de mesure électrochimique

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011099867A (ja) * 2002-07-31 2011-05-19 Toshiba Corp 塩基配列自動解析装置
JP2007514175A (ja) * 2003-12-15 2007-05-31 ジーンオーム サイエンシーズ、インク． 多重型電気化学的検出システムと方法
JP2009510446A (ja) * 2005-09-29 2009-03-12 コンビマトリックス・コーポレイション 電極マイクロアレイ上の結合事象の測定方法および装置
JP2010529007A (ja) * 2007-05-25 2010-08-26 ウニベルシダッド アウトノマ デ マドリッド 核酸配列の電気化学的検出法
JP2012047536A (ja) * 2010-08-25 2012-03-08 Nagoya Univ 電流検出装置
WO2017154801A1 (fr) * 2016-03-11 2017-09-14 パナソニックＩｐマネジメント株式会社 Système de mesure électrochimique, dispositif de mesure électrochimique, et procédé de mesure électrochimique

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