JP4434997B2 - Electrochemical measurement electrode, electrochemical measurement device, and electrochemical measurement method - Google Patents

Electrochemical measurement electrode, electrochemical measurement device, and electrochemical measurement method Download PDF

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JP4434997B2
JP4434997B2 JP2005063875A JP2005063875A JP4434997B2 JP 4434997 B2 JP4434997 B2 JP 4434997B2 JP 2005063875 A JP2005063875 A JP 2005063875A JP 2005063875 A JP2005063875 A JP 2005063875A JP 4434997 B2 JP4434997 B2 JP 4434997B2
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龍人 有村
和夫 橋口
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本発明は、電気化学測定用電極、電気化学測定用装置および電気化学測定方法に関する。   The present invention relates to an electrochemical measurement electrode, an electrochemical measurement device, and an electrochemical measurement method.

電気化学測定法は、液体試料中に電気化学測定用電極を浸漬し、液体試料中の物質が電気化学測定用電極に接触したときに起こる酸化反応または還元反応の際に電気化学測定用電極に流れる電流をモニタすることによって、液体試料中の物質を電気的に検出する方法である。   The electrochemical measurement method involves immersing an electrochemical measurement electrode in a liquid sample and applying it to the electrochemical measurement electrode during an oxidation reaction or reduction reaction that occurs when a substance in the liquid sample comes into contact with the electrochemical measurement electrode. This is a method of electrically detecting a substance in a liquid sample by monitoring a flowing current.

従来から、電気化学測定法は、水中、有機溶媒中または生体中などに含まれるイオンや分子の定性分析などに用いられている。このような電気化学測定法に用いられる電気化学測定用電極が、たとえば特許文献１に開示されている。この電気化学測定用電極は、絶縁性基板上に形成された複数の帯状の作用電極、対向電極および参照電極を有している。   Conventionally, electrochemical measurement methods have been used for qualitative analysis of ions and molecules contained in water, organic solvents, or living organisms. An electrode for electrochemical measurement used in such an electrochemical measurement method is disclosed in Patent Document 1, for example. This electrode for electrochemical measurements has a plurality of strip-like working electrodes, counter electrodes and reference electrodes formed on an insulating substrate.

図６の模式的な拡大平面図に、従来の電気化学測定用電極の作用電極部分の一例を示す。この電気化学測定用電極においては、互いに平行な一対のリード部９、１０と、各リード部から他方のリード部に向かって延び、それぞれ互い平行に１本ずつ交互に形成されている複数の帯状の作用電極１１、１２とが絶縁性基板７上に形成されている。このような電気化学測定用電極は、フォトリソグラフィ技術を利用した微細加工技術によって形成することができる。   An example of a working electrode portion of a conventional electrochemical measurement electrode is shown in the schematic enlarged plan view of FIG. In this electrochemical measurement electrode, a pair of lead portions 9 and 10 parallel to each other and a plurality of strips extending from each lead portion toward the other lead portion and alternately formed one by one in parallel with each other The working electrodes 11 and 12 are formed on the insulating substrate 7. Such an electrode for electrochemical measurement can be formed by a fine processing technique using a photolithography technique.

この電気化学測定用電極を液体試料中に浸し、作用電極１１と作用電極１２とにそれぞれ異なる電位を与えると、隣接する作用電極間で酸化還元サイクル（レドックスサイクル）が起こり、電流応答を電気的に増幅することができる。すなわち、液体試料中の物質が、たとえば作用電極１１で酸化された後に、その作用電極１１に隣接する作用電極１２で還元されるというサイクルが繰り返され、その結果、見かけの電流量を増大させることができる。したがって、電気化学測定用電極を用いた電気化学測定方法は、液体試料中の微量な物質の検出に好適に用いられる。   When this electrochemical measurement electrode is immersed in a liquid sample and different potentials are applied to the working electrode 11 and the working electrode 12, an oxidation-reduction cycle (redox cycle) occurs between adjacent working electrodes, and the current response is electrically Can be amplified. That is, a cycle in which a substance in a liquid sample is oxidized at, for example, the working electrode 11 and then reduced at the working electrode 12 adjacent to the working electrode 11 is repeated, and as a result, the apparent current amount is increased. Can do. Therefore, the electrochemical measurement method using the electrochemical measurement electrode is suitably used for detecting a very small amount of substance in a liquid sample.

また、電気化学測定用電極を用いた電気化学測定方法には、液体試料を強制対流させることなく電気化学測定が可能、ＩＲドロップが小さいおよび反応速度の速い反応の分析が可能といった特徴がある。   In addition, the electrochemical measurement method using the electrode for electrochemical measurement has characteristics such that electrochemical measurement can be performed without forced convection of a liquid sample, and a reaction with a small IR drop and a high reaction rate can be analyzed.

さらに、電気化学測定用電極を用いた電気化学測定方法は、抗原や抗体といったタンパク質の測定にも応用されている。たとえば、微量の抗原を検出する際は、抗原をマイクロビーズや基板に固定化された一次抗体と反応させて固定化し、その固定化した抗原に酵素標識された二次抗体を反応させ固定化する。その際、固定化される二次抗体は抗原の量に比例する。さらに、二次抗体の標識酵素に基質を反応させレドックス種を生成し、その生成したレドックス種を、レドックスサイクルを利用して測定することで、抗原の濃度を間接的に測定する競合法が用いられている。   Furthermore, the electrochemical measurement method using the electrochemical measurement electrode is also applied to the measurement of proteins such as antigens and antibodies. For example, when detecting a small amount of antigen, the antigen is reacted and immobilized with a primary antibody immobilized on microbeads or a substrate, and then the enzyme-labeled secondary antibody is reacted with and immobilized on the immobilized antigen. . At that time, the immobilized secondary antibody is proportional to the amount of the antigen. Furthermore, a competitive method that indirectly measures the concentration of the antigen is used by reacting the substrate with the labeling enzyme of the secondary antibody to generate a redox species, and measuring the generated redox species using a redox cycle. It has been.

また、図８に、特許文献２に記載されている電気化学測定用電極の模式的な斜視図を示す。この電気化学測定用電極においては、シリコン基板１４上に形成された、酸化膜１３、第２作用電極１５、二酸化シリコン膜１７および第１作用電極１８がこの順序で形成されており、第１作用電極１８の表面には複数の円形状の孔２０が配列されている。これらの孔２０は第１作用電極１８および二酸化シリコン膜１７を貫通しており、孔２０からは第２作用電極１５の表面が露出している。
特開平１−２７２９５８号公報 特許第２５６６１７３号公報 特開平３−２２１８５７号公報 FIG. 8 is a schematic perspective view of the electrode for electrochemical measurement described in Patent Document 2. In this electrochemical measurement electrode, the oxide film 13, the second working electrode 15, the silicon dioxide film 17, and the first working electrode 18 formed on the silicon substrate 14 are formed in this order, and the first working electrode is formed. A plurality of circular holes 20 are arranged on the surface of the electrode 18. These holes 20 penetrate the first working electrode 18 and the silicon dioxide film 17, and the surface of the second working electrode 15 is exposed from the hole 20.
JP-A-1-272958 Japanese Patent No. 2566173 JP-A-3-221857

しかしながら、上記の特許文献１および特許文献２に開示されている電気化学測定用電極においては感度が十分でないという問題があった。そこで、特許文献３においては、同一基板上に作用電極と作用電極電流増幅用のオペレーションアンプを形成することによって感度を向上させた電気化学測定用電極が開示されている。しかし、この電気化学測定用電極は、製造工程数が多いため、製造コストが高くなるという問題があった。   However, the electrochemical measurement electrodes disclosed in Patent Document 1 and Patent Document 2 have a problem that the sensitivity is not sufficient. Therefore, Patent Document 3 discloses an electrochemical measurement electrode in which sensitivity is improved by forming a working electrode and an operational amplifier for amplifying the working electrode current on the same substrate. However, this electrochemical measurement electrode has a problem that the manufacturing cost is high due to the large number of manufacturing steps.

本発明の目的は、感度を向上することができる電気化学測定用電極と、この電気化学測定用電極を含む電気化学測定用装置と、この電気化学測定用装置を用いた電気化学測定方法とを提供することにある。   An object of the present invention is to provide an electrochemical measurement electrode capable of improving sensitivity, an electrochemical measurement device including the electrochemical measurement electrode, and an electrochemical measurement method using the electrochemical measurement device. It is to provide.

本発明は、液体試料中の物質を電気的に検出する電気化学測定に用いられる電気化学測定用電極であって、絶縁性基板と、絶縁性基板上に設置された作用電極と、を含み、作用電極の周縁の少なくとも一部に凹凸が形成されており、前記作用電極は第１作用電極と第２作用電極とを含み、前記第１作用電極と前記第２作用電極とにそれぞれ異なる電位を与えることによって、前記第１作用電極で前記物質を酸化または還元することができ、前記第１作用電極で酸化または還元された前記物質を前記第２作用電極で還元または酸化することができることを特徴とする電気化学測定用電極である。 The present invention is an electrode for electrochemical measurement used for electrochemical measurement for electrically detecting a substance in a liquid sample, and includes an insulating substrate, and a working electrode placed on the insulating substrate, Concavities and convexities are formed on at least a part of the periphery of the working electrode, and the working electrode includes a first working electrode and a second working electrode, and different potentials are applied to the first working electrode and the second working electrode, respectively. by providing the first can be oxidized or reduced the material at the working electrode, the material that is oxidized or reduced at the first working electrode can be reduced or oxidized at the second working electrode Rukoto An electrode for electrochemical measurement characterized by the following.

また、本発明の電気化学測定用電極においては、第１作用電極と第２作用電極との間隔を２０ｎｍ以上１０μｍ以下とすることができる。   In the electrode for electrochemical measurements of the present invention, the distance between the first working electrode and the second working electrode can be set to 20 nm or more and 10 μm or less.

また、本発明は、液体試料中の物質を電気的に検出する電気化学測定に用いられる電気化学測定用電極であって、絶縁性基板と、絶縁性基板上において互いに向かい合うようにして設置されている第１作用電極と第２作用電極とを含み、第１作用電極には第１作用電極を貫通して第２作用電極の表面を露出する孔が形成され、孔の周縁の少なくとも一部に凹凸が形成されていることを特徴とする電気化学測定用電極である。   The present invention also relates to an electrode for electrochemical measurement used for electrochemical measurement for electrically detecting a substance in a liquid sample, and is installed so as to face each other on the insulating substrate and the insulating substrate. A first working electrode and a second working electrode, wherein the first working electrode is formed with a hole that penetrates the first working electrode and exposes a surface of the second working electrode, and is formed on at least a part of the periphery of the hole. It is an electrode for electrochemical measurements, characterized in that irregularities are formed.

ここで、本発明の電気化学測定用電極においては、第１作用電極と第２作用電極とにそれぞれ異なる電位を与えることによって、第１作用電極で物質を酸化または還元することができ、第１作用電極で酸化または還元された物質を第２作用電極で還元または酸化することができる。   Here, in the electrochemical measurement electrode of the present invention, the first working electrode can be oxidized or reduced by applying different potentials to the first working electrode and the second working electrode. Substances oxidized or reduced at the working electrode can be reduced or oxidized at the second working electrode.

また、本発明の電気化学測定用電極においては、絶縁性基板上に参照電極および対向電極の少なくとも一方が設置されていてもよい。   In the electrochemical measurement electrode of the present invention, at least one of a reference electrode and a counter electrode may be provided on the insulating substrate.

また、本発明の電気化学測定用電極において、凹凸は、山形状、波形状および方形状からなる群のうち少なくとも１種の形状からなることができる。   In the electrochemical measurement electrode of the present invention, the unevenness can be made of at least one shape selected from the group consisting of a mountain shape, a wave shape, and a square shape.

また、本発明は、上記のいずれかの電気化学測定用電極と、液体試料を収容するための容器と、を含む、電気化学測定用装置である。   In addition, the present invention is an electrochemical measurement apparatus including any one of the above electrochemical measurement electrodes and a container for storing a liquid sample.

さらに、本発明は、上記の電気化学測定用装置を用いて液体試料中の物質を電気的に検出する工程を含む電気化学測定方法である。   Furthermore, the present invention is an electrochemical measurement method including a step of electrically detecting a substance in a liquid sample using the above electrochemical measurement apparatus.

本発明によれば、感度を向上することができる電気化学測定用電極と、この電気化学測定用電極を含む電気化学測定用装置と、この電気化学測定用装置を用いた電気化学測定方法とを提供することができる。   According to the present invention, an electrochemical measurement electrode capable of improving sensitivity, an electrochemical measurement device including the electrochemical measurement electrode, and an electrochemical measurement method using the electrochemical measurement device are provided. Can be provided.

以下、本発明の実施の形態について説明する。なお、本発明の図面において、同一の参照符号は、同一部分または相当部分を表わすものとする。   Embodiments of the present invention will be described below. In the drawings of the present invention, the same reference numerals represent the same or corresponding parts.

図１に、本発明の電気化学測定用電極の好ましい一例の模式的な平面図を示す。本発明の電気化学測定用電極１は、絶縁性基板２上に形成された３本の帯状の第１作用電極５と３本の帯状の第２作用電極６とを含み、３本の第１作用電極５は導電性のリード部４によって電気的に接続され、３本の第２作用電極６は導電性のリード部３によって電気的に接続されている。そして、第１作用電極５の周縁の一部および第２作用電極６の周縁の一部にはそれぞれ凹凸が形成されている。第１作用電極５の長さＬ１と第２作用電極６の長さＬ１は、それぞれたとえば２０００μｍ以下とすることができる。また、第１作用電極５および第２作用電極６の形成本数は特に限定されず、第１作用電極５と第２作用電極６を１対としてたとえば５０対から１００対程度設置することもできる。また、第１作用電極５と第２作用電極６が形成された絶縁性基板と同一の絶縁性基板上に参照電極および対向電極の少なくとも一方を設置することもできる。   In FIG. 1, the typical top view of a preferable example of the electrode for electrochemical measurements of this invention is shown. The electrode for electrochemical measurement 1 of the present invention includes three strip-shaped first working electrodes 5 and three strip-shaped second working electrodes 6 formed on an insulating substrate 2, and includes three first electrodes. The working electrode 5 is electrically connected by the conductive lead 4, and the three second working electrodes 6 are electrically connected by the conductive lead 3. Unevenness is formed on a part of the periphery of the first working electrode 5 and a part of the periphery of the second working electrode 6. The length L1 of the first working electrode 5 and the length L1 of the second working electrode 6 can be set to 2000 μm or less, for example. The number of the first working electrode 5 and the second working electrode 6 formed is not particularly limited, and the first working electrode 5 and the second working electrode 6 can be installed as a pair, for example, about 50 to 100 pairs. Also, at least one of the reference electrode and the counter electrode can be provided on the same insulating substrate as the insulating substrate on which the first working electrode 5 and the second working electrode 6 are formed.

ここで、絶縁性基板２としては、たとえば表面に酸化膜などの絶縁性材料が形成されたシリコン基板、石英基板、酸化アルミニウム基板、ガラス基板またはプラスチック基板などを用いることができる。また、第１作用電極５および第２作用電極６は導電性材料から構成することができ、第１作用電極５および第２作用電極６を構成する導電性材料としては、たとえば金、白金、銀、クロム、チタンまたはステンレスなどを用いることができる。また、参照電極としては、たとえばＡｇ／ＡｇＣｌ電極、飽和カロメル電極または水素電極などを用いることができる。さらに、対向電極としては、たとえば白金電極またはカーボン電極などを用いることができる。   Here, as the insulating substrate 2, for example, a silicon substrate, a quartz substrate, an aluminum oxide substrate, a glass substrate, a plastic substrate, or the like on which an insulating material such as an oxide film is formed can be used. Moreover, the 1st working electrode 5 and the 2nd working electrode 6 can be comprised from an electroconductive material, As an electroconductive material which comprises the 1st working electrode 5 and the 2nd working electrode 6, it is gold, platinum, silver, for example Chrome, titanium, stainless steel, or the like can be used. As the reference electrode, for example, an Ag / AgCl electrode, a saturated calomel electrode, a hydrogen electrode, or the like can be used. Further, as the counter electrode, for example, a platinum electrode or a carbon electrode can be used.

図３に、図１に示す本発明の電気化学測定用電極１を用いた電気化学測定用装置の模式的な構成図を示す。図３に示す電気化学測定用装置において、容器３０には液体試料３１が収容されており、液体試料３１中には参照電極２１、電気化学測定用電極１および対向電極２５がそれぞれ浸漬させられている。そして、電気化学測定用電極１の第１作用電極５はリード部４を介してデュアル用ポテンショスタット２２に接続され、第２作用電極６はリード部３を介してポテンショスタット２３に接続されている。また、参照電極２１はデュアル用ポテンショスタット２２およびポテンショスタット２３に接続されており、対向電極２５はポテンショスタット２３に接続されている。さらに、ポテンショスタット２３はファンクションジェネレータ２４に接続されている。   FIG. 3 shows a schematic configuration diagram of an electrochemical measurement apparatus using the electrochemical measurement electrode 1 of the present invention shown in FIG. In the electrochemical measurement apparatus shown in FIG. 3, a liquid sample 31 is accommodated in the container 30, and the reference electrode 21, the electrochemical measurement electrode 1, and the counter electrode 25 are immersed in the liquid sample 31. Yes. The first working electrode 5 of the electrochemical measurement electrode 1 is connected to the dual potentiostat 22 via the lead 4, and the second working electrode 6 is connected to the potentiostat 23 via the lead 3. . The reference electrode 21 is connected to the dual potentiostat 22 and the potentiostat 23, and the counter electrode 25 is connected to the potentiostat 23. Further, the potentiostat 23 is connected to the function generator 24.

ここで、第１作用電極５と第２作用電極６とにそれぞれ異なる電位を与えることによって、電気化学測定用電極１の第１作用電極５と第２作用電極６との間で行なわれるレドックスサイクルを利用して液体試料３１中の物質を電気的に検出することができる。図３に示す電気化学測定装置においては、デュアル用ポテンショスタット２２によって第１作用電極５に与えられる電位を一定とし、ファンクションジェネレータ２４によって第２作用電極６に与えられる電位を変化させることによって、第１作用電極５と第２作用電極６とにそれぞれ異なる電位を与えることができる。   Here, a redox cycle performed between the first working electrode 5 and the second working electrode 6 of the electrochemical measurement electrode 1 by applying different potentials to the first working electrode 5 and the second working electrode 6, respectively. Can be used to electrically detect substances in the liquid sample 31. In the electrochemical measuring apparatus shown in FIG. 3, the potential applied to the first working electrode 5 by the dual potentiostat 22 is kept constant, and the potential applied to the second working electrode 6 is changed by the function generator 24 to change the first potential electrode 6. Different potentials can be applied to the first working electrode 5 and the second working electrode 6, respectively.

図７に、本発明の電気化学測定用電極におけるレドックスサイクルの一例を図解する模式的な断面図を示す。ここで、図７は図１のＶＩＩ−ＶＩＩに沿った模式的な断面図である。図７に示すように、たとえば液体試料中の物質であるＲｅｄは第１作用電極５によって酸化されてＯｘとなった後に第１作用電極５に隣接する第２作用電極６によって再度還元されて元のＲｅｄに戻る。このとき、第１作用電極５および第２作用電極６にそれぞれ電流が流れ、この電流によって液体試料中の物質が電気的に検知される。これは、液体試料中の物質が第１作用電極５によって還元された後に第１作用電極５に隣接する第２作用電極６によって酸化される場合も同様である。そして、単位時間当たりのレドックスサイクル数が多いほど、単位時間当たりに第１作用電極５および第２作用電極６に流れる電流量が増加するため感度が高くなる。   FIG. 7 is a schematic cross-sectional view illustrating an example of the redox cycle in the electrochemical measurement electrode of the present invention. Here, FIG. 7 is a schematic cross-sectional view taken along the line VII-VII in FIG. As shown in FIG. 7, for example, Red, which is a substance in the liquid sample, is oxidized by the first working electrode 5 to become Ox and then reduced again by the second working electrode 6 adjacent to the first working electrode 5. Return to Red. At this time, currents flow through the first working electrode 5 and the second working electrode 6, respectively, and substances in the liquid sample are electrically detected by the currents. The same applies to the case where the substance in the liquid sample is oxidized by the second working electrode 6 adjacent to the first working electrode 5 after being reduced by the first working electrode 5. As the number of redox cycles per unit time increases, the amount of current flowing through the first working electrode 5 and the second working electrode 6 per unit time increases, and thus the sensitivity increases.

本発明においては、第１作用電極５および第２作用電極６のそれぞれの周縁に凹凸が形成されていることから、単位時間当たりのレドックスサイクル数を増加することができるため感度をより向上することができる。   In the present invention, since the unevenness is formed on the periphery of each of the first working electrode 5 and the second working electrode 6, the number of redox cycles per unit time can be increased, so that the sensitivity is further improved. Can do.

すなわち、本発明においては、作用電極の周縁の少なくとも一部に凹凸を形成することによって、周縁に凹凸が形成されていない従来の作用電極と比べて液体試料中の物質の接触領域を広げることができるため、作用電極の周縁部でより多くの酸化物質または還元物質（レドックス種）を生成することができる。これにより、隣接する作用電極との間におけるレドックス種の濃度差をより大きなものとすることができるため、隣接する作用電極へのレドックス種の拡散をより促進することができる。さらに、本発明において、上記凹凸は隣接する作用電極への到達時間の最も短い作用電極の周縁に形成されている。したがって、本発明においては、作用電極の周縁の少なくとも一部に凹凸を形成することによって、隣接する作用電極へのレドックス種の拡散効率が向上し、単位時間当たりのレドックスサイクル数を従来よりも大幅に増加することができるため、感度を向上することができるのである。   That is, in the present invention, by forming irregularities on at least a part of the periphery of the working electrode, it is possible to widen the contact area of the substance in the liquid sample as compared with the conventional working electrode having no irregularities on the peripheral edge. Therefore, more oxidizing substance or reducing substance (redox species) can be generated at the peripheral edge of the working electrode. Thereby, since the density | concentration difference of the redox seed | species between adjacent working electrodes can be made larger, the spreading | diffusion of the redox seed | species to the adjacent working electrode can be promoted more. Furthermore, in the present invention, the unevenness is formed on the periphery of the working electrode having the shortest arrival time to the adjacent working electrode. Therefore, in the present invention, by forming irregularities on at least a part of the peripheral edge of the working electrode, the diffusion efficiency of the redox species to the adjacent working electrode is improved, and the number of redox cycles per unit time is significantly larger than before. Therefore, the sensitivity can be improved.

図２に、図１の破線Ａで囲まれた部分の模式的な拡大平面図を示す。図２に示すように、第１作用電極５の周縁および第２作用電極６の周縁にはそれぞれ山形状の凹凸が形成されている。ここで、第１作用電極５と第２作用電極６との間隔Ｇ１が２０ｎｍ以上１０μｍ以下であることが好ましい。これは、本発明者が鋭意検討した結果、間隔Ｇ１が２０ｎｍ以上１０μｍ以下である場合には、第１作用電極５および第２作用電極６のそれぞれに流れる電流量が増加し、感度をより向上することができる傾向にあることを見出したことによるものである。ここで、図２に示す間隔Ｇ１は、第１作用電極５と第２作用電極６の隣接方向における、第１作用電極５の電極外側への最突出部と電極内側への最突出部との中点と、第２作用電極６の電極外側への最突出部と電極内側への最突出部との中点との間の距離を示している。また、感度をさらに高めることができ、第１作用電極５および第２作用電極６の形状を大きく変化させないという観点からは、第１作用電極５および第２作用電極６の凹凸の高さＷ２は第１作用電極５および第２作用電極６の幅Ｗ１の１／２以下とすることが好ましい。第１作用電極５および第２作用電極６の幅Ｗ１はたとえば２０ｎｍ以上１０μｍ以下とすることができる。また、第１作用電極５および第２作用電極６の厚さはたとえば１０ｎｍ以上１００ｎｍ以下とすることができる。   FIG. 2 is a schematic enlarged plan view of a portion surrounded by a broken line A in FIG. As shown in FIG. 2, mountain-shaped irregularities are formed on the periphery of the first working electrode 5 and the periphery of the second working electrode 6, respectively. Here, the gap G1 between the first working electrode 5 and the second working electrode 6 is preferably 20 nm or more and 10 μm or less. As a result of intensive studies by the present inventors, when the gap G1 is 20 nm or more and 10 μm or less, the amount of current flowing through each of the first working electrode 5 and the second working electrode 6 is increased, and the sensitivity is further improved. This is because they found out that they tend to be able to. Here, the gap G1 shown in FIG. 2 is the distance between the most projecting portion on the outer side of the first working electrode 5 and the most projecting portion on the inner side of the electrode in the adjacent direction of the first working electrode 5 and the second working electrode 6. The distance between the midpoint and the midpoint between the outermost protruding portion of the second working electrode 6 and the outermost protruding portion of the second working electrode 6 is shown. Further, from the viewpoint that the sensitivity can be further increased and the shapes of the first working electrode 5 and the second working electrode 6 are not greatly changed, the unevenness height W2 of the first working electrode 5 and the second working electrode 6 is It is preferable that the width W1 of the first working electrode 5 and the second working electrode 6 is ½ or less. The width W1 of the first working electrode 5 and the second working electrode 6 can be set to, for example, 20 nm or more and 10 μm or less. Moreover, the thickness of the 1st working electrode 5 and the 2nd working electrode 6 can be 10 nm or more and 100 nm or less, for example.

また、第１作用電極５および第２作用電極６の凹凸の形状は、たとえば図４の模式的な拡大平面図を示すような波形状とすることもでき、たとえば図５の模式的な拡大平面図を示すような方形状とすることもできる。   Moreover, the uneven | corrugated shape of the 1st working electrode 5 and the 2nd working electrode 6 can also be made into a wave shape which shows the typical enlarged plan view of FIG. 4, for example, for example, the typical enlarged plane of FIG. It can also be a square shape as shown in the figure.

図１に示す本発明の電気化学測定用電極１は、たとえばフォトリソグラフィ技術を利用して作製することができる。すなわち、たとえば、まず、絶縁性基板２上に、周縁に凹凸を有する第１作用電極５および第２作用電極６の形状に開口部が形成されたレジスト膜を設置する。次に、このレジスト膜の開口部に第１作用電極５および第２作用電極６を構成する導電性材料を堆積する。最後に、レジスト膜を除去することによって、図１に示す電気化学測定用電極１を作製することができる。   The electrochemical measurement electrode 1 of the present invention shown in FIG. 1 can be produced by using, for example, a photolithography technique. That is, for example, first, a resist film in which openings are formed in the shape of the first working electrode 5 and the second working electrode 6 having irregularities on the periphery is placed on the insulating substrate 2. Next, a conductive material constituting the first working electrode 5 and the second working electrode 6 is deposited in the opening of the resist film. Finally, the electrochemical measurement electrode 1 shown in FIG. 1 can be produced by removing the resist film.

図１０に、本発明の電気化学測定用電極の好ましい一例の模式的な斜視図を示す。この電気化学測定用電極１においては、絶縁性基板２上には絶縁膜２６、第２作用電極６、絶縁膜２７および第１作用電極５がこの順序で形成されている。ここで、絶縁性基板２上において第１作用電極５と第２作用電極６とは互いに向かい合うようにして設置され、第１作用電極５には第１作用電極５を貫通して第２作用電極６の表面を露出する孔２０が形成されており、孔２０の周縁の少なくとも一部に凹凸が形成されていることに特徴がある。この場合にも、孔２０の周縁に形成されている凹凸よって液体試料中の物質の接触領域を広くすることができるため、孔２０の周縁部でより多くのレドックス種を生成することができ、第２作用電極６との間におけるレドックス種の濃度差をより大きなものとして隣接する第２作用電極６へのレドックス種の拡散をより促進することができる。したがって、この場合にも、レドックス種の拡散効率が向上し、単位時間当たりのレドックスサイクル数を従来よりも大幅に増加することができるため、感度を向上することができる。   In FIG. 10, the typical perspective view of a preferable example of the electrode for electrochemical measurements of this invention is shown. In this electrochemical measurement electrode 1, an insulating film 26, a second working electrode 6, an insulating film 27 and a first working electrode 5 are formed in this order on the insulating substrate 2. Here, the first working electrode 5 and the second working electrode 6 are installed on the insulating substrate 2 so as to face each other, and the first working electrode 5 penetrates the first working electrode 5 and the second working electrode. A hole 20 exposing the surface of the hole 6 is formed, and unevenness is formed on at least a part of the periphery of the hole 20. Also in this case, since the contact area of the substance in the liquid sample can be widened by the unevenness formed at the periphery of the hole 20, more redox species can be generated at the periphery of the hole 20, It is possible to further promote the diffusion of the redox species to the adjacent second working electrode 6 by setting the concentration difference of the redox species between the second working electrode 6 to be larger. Accordingly, also in this case, the redox species diffusion efficiency is improved, and the number of redox cycles per unit time can be significantly increased as compared with the conventional case, so that the sensitivity can be improved.

図９に、図１０の破線Ｂで囲まれた部分の模式的な拡大平面図を示す。図９に示すように、孔２０の周縁にはそれぞれ山形状の凹凸が形成されている。ここで、孔２０の幅Ｗ１０は１０ｎｍ以上１０μｍ以下であることが好ましい。また、感度をさらに高めることができ、孔２０の形状を大きく変化させない観点から、凹凸の高さＷ１１および幅Ｗ１２は孔２０の幅Ｗ１０の１／１０以下とすることが好ましい。   FIG. 9 shows a schematic enlarged plan view of a portion surrounded by a broken line B in FIG. As shown in FIG. 9, mountain-shaped irregularities are formed on the periphery of the hole 20. Here, the width W10 of the hole 20 is preferably 10 nm or more and 10 μm or less. Moreover, it is preferable that the height W11 and the width W12 of the unevenness be 1/10 or less of the width W10 of the hole 20 from the viewpoint that sensitivity can be further increased and the shape of the hole 20 is not greatly changed.

図１０に示す本発明の電気化学測定用電極１も、たとえばフォトリソグラフィ技術を利用して作製することができる。すなわち、たとえば、まず、絶縁性基板２上に、絶縁膜２６、第２作用電極６、絶縁膜２７および第１作用電極５を構成する導電性材料をこの順序で形成する。次に、周縁に凹凸を有する孔２０の形状に開口部が形成されたレジスト膜を設置する。続いて、第１作用電極５を構成する導電性材料をレジスト膜の開口部の形状にエッチングして除去することによって、第２作用電極６の表面を露出させる。最後に、レジスト膜を除去することによって、図１０に示す電気化学測定用電極１を作製することができる。   The electrochemical measurement electrode 1 of the present invention shown in FIG. 10 can also be produced using, for example, photolithography. That is, for example, first, the conductive material constituting the insulating film 26, the second working electrode 6, the insulating film 27, and the first working electrode 5 is formed on the insulating substrate 2 in this order. Next, a resist film in which openings are formed in the shape of the holes 20 having irregularities on the periphery is provided. Subsequently, the surface of the second working electrode 6 is exposed by etching away the conductive material constituting the first working electrode 5 into the shape of the opening of the resist film. Finally, the electrochemical measurement electrode 1 shown in FIG. 10 can be produced by removing the resist film.

今回開示された実施の形態はすべての点で例示であって制限的なものではないと考えられるべきである。本発明の範囲は上記した説明ではなくて特許請求の範囲によって示され、特許請求の範囲と均等の意味および範囲内でのすべての変更が含まれることが意図される。   The embodiment disclosed this time should be considered as illustrative in all points and not restrictive. The scope of the present invention is defined by the terms of the claims, rather than the description above, and is intended to include any modifications within the scope and meaning equivalent to the terms of the claims.

本発明の電気化学測定用電極は、たとえばバイオセンサ、アレルゲンセンサ、ＤＮＡチップまたはμＴＡＳ（マイクロ総合分析システム）などに好適に利用することができる。   The electrochemical measurement electrode of the present invention can be suitably used for, for example, a biosensor, an allergen sensor, a DNA chip, or μTAS (micro total analysis system).

本発明の電気化学測定用電極の好ましい一例の模式的な平面図である。1 is a schematic plan view of a preferred example of an electrode for electrochemical measurement according to the present invention. 図１の破線Ａで囲まれた部分の模式的な拡大平面図である。FIG. 2 is a schematic enlarged plan view of a portion surrounded by a broken line A in FIG. 1. 図１に示す本発明の電気化学測定用電極を用いた電気化学測定用装置の模式的な構成図である。It is a typical block diagram of the apparatus for electrochemical measurements using the electrode for electrochemical measurements of this invention shown in FIG. 本発明の電気化学測定用電極の作用電極の凹凸の形状の他の好ましい一例を示す模式的な拡大平面図である。It is a typical enlarged plan view which shows another preferable example of the uneven | corrugated shape of the working electrode of the electrode for electrochemical measurements of this invention. 本発明の電気化学測定用電極の作用電極の凹凸の形状のさらに他の好ましい一例を示す模式的な拡大平面図である。It is a typical enlarged plan view which shows another preferable example of the uneven | corrugated shape of the working electrode of the electrode for electrochemical measurements of this invention. 従来の電気化学測定用電極の作用電極部分の一例を示す模式的な拡大平面図である。It is a typical enlarged plan view which shows an example of the working electrode part of the conventional electrode for electrochemical measurements. 本発明の電気化学測定用電極におけるレドックスサイクルの一例を図解する模式的な断面図である。It is typical sectional drawing illustrating an example of the redox cycle in the electrode for electrochemical measurements of this invention. 従来の電気化学測定用電極の模式的な斜視図である。It is a typical perspective view of the conventional electrode for electrochemical measurements. 図１０の破線Ｂで囲まれた部分の模式的な拡大平面図である。FIG. 11 is a schematic enlarged plan view of a portion surrounded by a broken line B in FIG. 10. 本発明の電気化学測定用電極の好ましい一例の模式的な斜視図である。It is a typical perspective view of a preferable example of the electrode for electrochemical measurements of the present invention.

符号の説明Explanation of symbols

１ 電気化学測定用電極、２，７ 絶縁性基板、３，４，９，１０ リード部、５，１８ 第１作用電極、６，１５ 第２作用電極、１１，１２ 作用電極、１３ 酸化膜、１４ シリコン基板、１７ 二酸化シリコン膜、２０ 孔、２１ 参照電極、２２ デュアル用ポテンショスタット、２３ ポテンショスタット、２４ ファンクションジェネレータ、２５ 対向電極、２６，２７ 絶縁膜、３０ 容器、３１ 液体試料。   1 Electrochemical measurement electrode, 2, 7 Insulating substrate, 3, 4, 9, 10 Lead part, 5, 18 First working electrode, 6, 15 Second working electrode, 11, 12 Working electrode, 13 Oxide film, 14 silicon substrate, 17 silicon dioxide film, 20 holes, 21 reference electrode, 22 potentiostat for dual, 23 potentiostat, 24 function generator, 25 counter electrode, 26, 27 insulating film, 30 container, 31 liquid sample.

Claims (8)

液体試料中の物質を電気的に検出する電気化学測定に用いられる電気化学測定用電極であって、絶縁性基板と、前記絶縁性基板上に設置された作用電極と、を含み、前記作用電極の周縁の少なくとも一部に凹凸が形成されており、前記作用電極は第１作用電極と第２作用電極とを含み、前記第１作用電極と前記第２作用電極とにそれぞれ異なる電位を与えることによって、前記第１作用電極で前記物質を酸化または還元することができ、前記第１作用電極で酸化または還元された前記物質を前記第２作用電極で還元または酸化することができることを特徴とする、電気化学測定用電極。 An electrode for electrochemical measurement used for electrochemical measurement for electrically detecting a substance in a liquid sample, comprising: an insulating substrate; and a working electrode placed on the insulating substrate, the working electrode Concavities and convexities are formed on at least a part of the peripheral edge, and the working electrode includes a first working electrode and a second working electrode, and applies different potentials to the first working electrode and the second working electrode, respectively. Accordingly, the first can be oxidized or reduced the material at the working electrode, the said Rukoto can oxidized or reduced the material in the first working electrode is reduced or oxidized at the second working electrode An electrode for electrochemical measurement. 前記第１作用電極と前記第２作用電極との間隔が２０ｎｍ以上１０μｍ以下であることを特徴とする、請求項１に記載の電気化学測定用電極。 The electrode for electrochemical measurements according to claim 1 , wherein a distance between the first working electrode and the second working electrode is 20 nm or more and 10 µm or less. 液体試料中の物質を電気的に検出する電気化学測定に用いられる電気化学測定用電極であって、絶縁性基板と、前記絶縁性基板上において互いに向かい合うようにして設置されている第１作用電極と第２作用電極とを含み、前記第１作用電極には前記第１作用電極を貫通して前記第２作用電極の表面を露出する孔が形成され、前記孔の周縁の少なくとも一部に凹凸が形成されていることを特徴とする、電気化学測定用電極。   An electrode for electrochemical measurement used for electrochemical measurement for electrically detecting a substance in a liquid sample, the insulating substrate and a first working electrode installed on the insulating substrate so as to face each other And a second working electrode, wherein the first working electrode is formed with a hole that penetrates the first working electrode and exposes a surface of the second working electrode, and at least part of a peripheral edge of the hole is uneven. An electrode for electrochemical measurement, characterized in that is formed. 前記第１作用電極と前記第２作用電極とにそれぞれ異なる電位を与えることによって、前記第１作用電極で前記物質を酸化または還元することができ、前記第１作用電極で酸化または還元された前記物質を前記第２作用電極で還元または酸化することができることを特徴とする、請求項３に記載の電気化学測定用電極。 By applying different potentials to the first working electrode and the second working electrode, the substance can be oxidized or reduced by the first working electrode, and the substance oxidized or reduced by the first working electrode. The electrode for electrochemical measurement according to claim 3 , wherein a substance can be reduced or oxidized by the second working electrode. 前記絶縁性基板上に参照電極および対向電極の少なくとも一方が設置されていることを特徴とする、請求項１から４のいずれかに記載の電気化学測定用電極。 The electrode for electrochemical measurement according to any one of claims 1 to 4 , wherein at least one of a reference electrode and a counter electrode is provided on the insulating substrate. 前記凹凸は、山形状、波形状および方形状からなる群のうち少なくとも１種の形状からなることを特徴とする、請求項１から５のいずれかに記載の電気化学測定用電極。 The electrode for electrochemical measurements according to any one of claims 1 to 5 , wherein the unevenness comprises at least one shape selected from the group consisting of a mountain shape, a wave shape, and a square shape. 請求項１から６のいずれかに記載の電気化学測定用電極と、前記液体試料を収容するための容器と、を含む、電気化学測定用装置。 An electrochemical measuring electrode according to any one of claims 1 to 6, including a container for containing the liquid sample, the electrochemical measurement device. 請求項７に記載の電気化学測定用装置を用いて前記液体試料中の前記物質を電気的に検出する工程を含む、電気化学測定方法。 An electrochemical measurement method comprising a step of electrically detecting the substance in the liquid sample using the electrochemical measurement device according to claim 7 .
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