WO2023122894A1 - 生物传感芯片及其电极活化方法和装置 - Google Patents

生物传感芯片及其电极活化方法和装置 Download PDF

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WO2023122894A1
WO2023122894A1 PCT/CN2021/141710 CN2021141710W WO2023122894A1 WO 2023122894 A1 WO2023122894 A1 WO 2023122894A1 CN 2021141710 W CN2021141710 W CN 2021141710W WO 2023122894 A1 WO2023122894 A1 WO 2023122894A1
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electrode
activation
working electrode
activation method
voltage signal
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PCT/CN2021/141710
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French (fr)
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陈玮
张宇宁
黎宇翔
章朦
朱叶宁
云全新
董宇亮
章文蔚
徐讯
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深圳华大生命科学研究院
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Priority to PCT/CN2021/141710 priority Critical patent/WO2023122894A1/zh
Publication of WO2023122894A1 publication Critical patent/WO2023122894A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells

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  • the present application relates to the field of electrochemical technology, in particular to a biosensing chip and its electrode activation method and device.
  • the branch of electrochemical discipline is divided into batteries (such as lithium batteries), supercapacitors and biosensing, among which electrochemical biosensing plays an important role in medical devices.
  • batteries such as lithium batteries
  • supercapacitors and biosensing
  • electrochemical biosensing plays an important role in medical devices.
  • various methodologies compete for market share.
  • Mainstream platforms include chemiluminescence (electrochemiluminescence), specific proteins, biochemistry, immunity, blood coagulation, blood cells, blood gas, colloidal gold, molecular diagnosis, POCT (point of care testing, instant inspection), etc.
  • Electrochemical biosensing involves The projects include fixed frequency AC impedance blood coagulation chip, current/voltage/conductivity dry blood gas biochemical chip/wet electrode, first, second and third generation biosensor blood glucose test strips/three types of medical device implantable continuous monitoring blood glucose electrode and high-end gene sequencing -Nanopore electrochemical single-molecule gene sequencing chips, etc.
  • the substrate chips used in electrochemical biosensing generally include screen-printed electrodes (SPE), printed circuit boards (PCB), flexible circuit boards (FPCB) and semiconductor (Semi-conductor) chips; Screen printing process, PCB process and MEMS (Micro Electro Mechanical System) process.
  • SPE screen-printed electrodes
  • PCB printed circuit boards
  • FPCB flexible circuit boards
  • semiconductor Semi-conductor
  • the electrochemical biosensing chip is most important in addition to the electrode active area-reactive area, it is also particularly important to maintain the hydrophilicity and hydrophobicity of the inert area of the protective lead or reaction tank.
  • the plasma such as oxygen, argon, helium and nitrogen, etc.
  • the use of silanized hydrophobic property refolding will also lead to the same modification of the active area of the chip electrode, directly causing poor conduction.
  • the matrix electrode or high-end biosensing chip manufacturing process achieves higher electrode integration and smaller area, such as micro-nano processing technology, which is applied to the development stage of electrochemical biosensing product projects to a great extent.
  • There will be fatal defects such as electrical disconnection or poor performance.
  • Simple physical polishing, cutting, cleaning and scouring or chemical immersion cannot effectively solve the problem.
  • the main purpose of the present application is to provide a biosensor chip and its electrode activation method and device, so as to improve the above-mentioned defects in the prior art.
  • the biosensor chip includes a carrier plate, a working electrode and a counter electrode, and an inert region and the working electrode are respectively formed on the carrier plate. an electrode, the working electrode having an electrocatalytic substance;
  • the electrode activation method comprises the following steps:
  • controlling the signal generator to apply an activation electrical signal to the biosensing chip to activate the working electrode and/or the counter electrode through the electrolyte, wherein the potential of the activation electrical signal is lower than that of the electrocatalytic substance oxidation potential.
  • the working electrode includes at least one working electrode unit.
  • the working electrode includes at least two working electrode units distributed in a periodic array.
  • the number ratio of the counter electrode to the working electrode unit ranges from 1:16 to 1:4096.
  • each of the working electrode units has an electrocatalytic substance.
  • the electrocatalytic substance includes any one or more of graphene, noble metal, colloidal gold or a composite of the aforementioned substances.
  • the activation electrical signal includes a DC voltage signal, an AC voltage signal or a DC voltage signal superimposed with an AC voltage signal.
  • the DC voltage signal when the activation electrical signal includes a DC voltage signal, the DC voltage signal includes a stepped DC voltage signal.
  • the scanning speed range of the stepped DC voltage signal is 0.001V/s ⁇ 1.0V/s.
  • the activation electrical signal when the activation electrical signal includes an AC voltage signal or a DC voltage signal superimposed on an AC voltage signal, the AC voltage signal includes a sine wave, a triangle wave, a sawtooth wave, a square wave, and a preset wave AC voltage signal any one or more of them.
  • the frequency of the AC voltage signal is 0.1 Hz ⁇ 1 GHz.
  • the frequency of the AC voltage signal is 1 Hz ⁇ 1 KHz.
  • the direction of the activation electrical signal includes any one or more of unidirectional oxidation, unidirectional reduction, and bidirectional symmetric redox.
  • the potential range of the activation electrical signal is -1.2V ⁇ 1.8V.
  • the total time period for applying the activation electrical signal ranges from 1 to 100 activation reaction cycles.
  • the total time period for applying the activation electrical signal ranges from 10 to 50 activation reaction cycles.
  • the electrolyte contains sacrificial reactants, and the sacrificial reactants include one or more of acids, bases, oxides or reducing substances.
  • the aqueous solution of the sacrificial reactant has a molar concentration ranging from 0.1 mM to 500 mM.
  • the inert material in the inert area includes any one of SU-8 (photoresist), epoxy resin, polyimide, FR4 (code designation for flame-resistant material grade) board and glass fiber layer one or more species.
  • SU-8 photoresist
  • epoxy resin epoxy resin
  • polyimide polyimide
  • FR4 code designation for flame-resistant material grade
  • a biosensing chip which uses the electrode activation method of the biosensing chip as described above to perform electrode activation.
  • an electrode activation device for a biosensor chip includes a carrier plate, a working electrode and a counter electrode, and an inert region and the working electrode are respectively formed on the carrier plate. an electrode, the working electrode having an electrocatalytic substance;
  • the electrode activation device includes:
  • a container for containing an electrolyte solution When the electrode is activated, the biosensing chip is placed in the container containing the electrolyte solution, so that the working electrode and the counter electrode are respectively immersed in the in the electrolyte; and,
  • a signal generator configured to apply an activation electrical signal to the biosensing chip to activate the working electrode and/or the counter electrode through the electrolyte, wherein the activation electrical signal has a potential lower than that of the electrical activation signal The oxidation potential of the catalytic species.
  • the biosensing chip and its electrode activation method and device provided by the application enhance the electrochemical performance of the micro-nano biosensing chip, greatly reduce the charge transfer resistance and increase the active effective area, and can control the inter-batch/intra-batch performance of the chip. Electrochemical difference, reducing the coefficient of variation of performance, can effectively ensure the same electrochemical quality of the chip before use, and can partially treat the reaction area, without affecting the hydrophilic and hydrophobic properties of the inert area, especially for the pretreatment of high-end chips of Class II and Class III medical devices
  • the usage specification provides a reference, which is beneficial to the research and development of biosensing chips.
  • FIG. 1 is a schematic flowchart of an electrode activation method of a biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 2 is a partial structural schematic diagram of a biosensing chip and an electrode activation device thereof according to an exemplary embodiment of the present application.
  • FIG. 3A is a schematic diagram of a first type of activation electrical signal applied by a signal generator according to an exemplary embodiment of the present application.
  • FIG. 3B is a schematic diagram of a second type of activation electrical signal applied by a signal generator according to an exemplary embodiment of the present application.
  • FIG. 3C is a schematic diagram of a third type of activation electrical signal applied by a signal generator according to an exemplary embodiment of the present application.
  • FIG. 3D is a schematic diagram of a fourth type of activation electrical signal applied by the signal generator according to an exemplary embodiment of the present application.
  • FIG. 3E is a schematic diagram of a fifth type of activation electrical signal applied by the signal generator according to an exemplary embodiment of the present application.
  • FIG. 3F is a schematic diagram of a sixth type of activation electrical signal applied by a signal generator according to an exemplary embodiment of the present application.
  • FIG. 4 is a schematic diagram showing a comparison of contact angles of inert regions before and after activation by an electrode activation method of a biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 5 is a schematic diagram of scanning speed image comparison before and after the activation method of the electrode of the biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 6 is a schematic diagram of comparison of multi-sweep cyclic voltammetry before and after activation by the electrode activation method of the biosensing chip according to an exemplary embodiment of the present application.
  • Fig. 7 is a schematic diagram of comparing open circuit potentials before and after activation by the electrode activation method of the biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 8 is a schematic diagram of the comparison of AC impedance before and after activation by the electrode activation method of the biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 9 is a three-dimensional schematic diagram of impedance before and after activation by the electrode activation method of the biosensing chip according to an exemplary embodiment of the present application.
  • FIG. 10 is a schematic diagram of equivalent circuit fitting of an electrode activation method of a biosensing chip according to an exemplary embodiment of the present application.
  • references in the specification to "an embodiment,” “an alternative embodiment,” “another embodiment,” etc. indicate that the described embodiments may include a particular feature, structure, or characteristic, but each embodiment The specific feature, structure or characteristic may not necessarily be included. Moreover, such phrases are not necessarily referring to the same embodiment. Furthermore, when a particular feature, structure or characteristic is described in conjunction with an embodiment, it is within the purview of those skilled in the relevant arts to implement such feature, structure or characteristic in conjunction with other embodiments, whether or not explicitly described.
  • first and second are used for descriptive purposes only, and cannot be interpreted as indicating or implying relative importance or implicitly specifying the quantity of indicated technical features. Thus, a feature defined as “first” and “second” may explicitly or implicitly include one or more of these features.
  • plural means two or more.
  • the term “comprise” and any variations thereof, are intended to cover a non-exclusive inclusion.
  • connection should be understood in a broad sense, for example, it can be a fixed connection or a flexible connection.
  • Detachable connection, or integral connection it can be mechanical connection or electrical connection; it can be direct connection or indirect connection through an intermediary, and it can be the internal communication of two components.
  • the present embodiment provides a method for activating electrodes of a biosensing chip.
  • the biosensing chip includes a carrier plate, a working electrode and a counter electrode, and an inert region and a working electrode are respectively formed on the carrier plate.
  • the electrode has an electrocatalytic substance; the electrode activation method includes the following steps: placing the biosensing chip in a container containing an electrolyte, so that the working electrode and the counter electrode are respectively immersed in the electrolyte; Applying an activation electrical signal to activate the working electrode and/or the counter electrode through the electrolyte, wherein the activation electrical signal has a potential less than the oxidation potential of the electrocatalytic species.
  • the electrode activation method in addition to being preferably applied in the field of biosensor chips, can also be applied to other fields such as semiconductors, high-end manufacturing, and other medical devices. This embodiment does not specifically limit the electrode activation method.
  • the application fields can be selected and adjusted according to actual needs or possible needs.
  • the electrodes of the biosensing chip adopt a current loop two-electrode system of 1 pair n (n is a natural number greater than or equal to 1), and no external reference electrode is needed, wherein the counter electrode can be a macro electrode, and the working electrode It can be a micro-matrix electrode, but the type of electrode is not specifically limited, and can be selected and adjusted according to actual needs or possible needs.
  • the electrocatalytic substance is not consumed or stripped during the electroactivation process of the working electrode and/or the counter electrode, and at the same time, the active area of the counter electrode and the working electrode can be greatly increased;
  • the electrode activation method Apply an activation electrical signal to the biosensing chip through a signal generator using an electromotive force (EMF), and the application of a potential range will not passivate the active reaction layer of the macro/micro matrix electrode, but will oxidize or reduce the inert substance on the electrode surface, Accelerates cleaning while activating.
  • EMF electromotive force
  • the electrode activation method of the biosensor chip mainly includes the following steps:
  • Step 101 providing a biosensor chip to be activated.
  • biosensing chip provided in this step is described below, but the biosensing chip should not be limited to the structure described below, and can be selected and adjusted according to actual needs or possible needs.
  • the biosensor chip mainly includes a carrier plate (not shown), a counter electrode 21 and a working electrode 22, an inert region 24 and a working electrode 22 are respectively formed on the carrier plate, and the working electrode 22 has an electrocatalytic function.
  • Substance 23 Specifically, the surface of the working electrode 22 supports electrocatalytic substances.
  • the counter electrode 21 may be a macro electrode of a biosensor chip
  • the working electrode 22 may include at least one working electrode unit 221 .
  • the working electrode 22 can be formed by at least two working electrode units 221 distributed in a periodic array, that is, the working electrode 22 can be a micro-matrix electrode, but the number and number of working electrode units 221 are not specifically limited.
  • the shape of the distribution can be selected and adjusted according to actual needs or possible needs.
  • the number ratio of the counter electrode 21 to the working electrode unit 221 ranges from 1:16 to 1:4096, that is, when the biosensor chip includes one counter electrode 21, the biosensor chip can include 16 to 4096
  • the number of working electrode units 221 is of course not limited to this number ratio range.
  • the number ratio of the counter electrode 21 to the working electrode unit 221 is 1:256.
  • each working electrode unit 221 has a corresponding electrocatalytic substance 23 .
  • the electrocatalytic substance 23 may include any one or more of graphene, noble metal, colloidal gold, or a composite of the aforementioned substances, and of course it is not limited thereto. Select and adjust accordingly.
  • the graphene may be three-dimensional graphene
  • the gold may be roughened gold
  • the noble metal may be a noble metal layer or particles
  • the noble metal may specifically be platinum or palladium.
  • the electrocatalytic substance 23 can be combined to the working electrode unit 221 by any one or more methods of electrochemical deposition, chemical deposition, valve dispensing and physical vapor deposition, but it is certainly not limited thereto.
  • physical vapor deposition may be magnetron sputtering.
  • the electrocatalytic substance can reduce the theoretically required voltage value of the electrolysis reaction and reduce the overpotential.
  • the inert material in the inert area 24 can include any one or more of SU-8, epoxy resin, polyimide, FR4 board and glass fiber layer, and of course it is not limited thereto. Make corresponding selections and adjustments based on actual needs or possible needs.
  • the inert substance may be SU-8.
  • Step 102 placing the biosensing chip in the container containing the electrolyte, so that the working electrode and the counter electrode are respectively immersed in the electrolyte.
  • the biosensor chip provided in step 101 is placed in the container 31 containing the electrolyte 32 , so that each working electrode unit 221 and the counter electrode 21 are soaked in the electrolyte 32 respectively.
  • the electrolyte 32 contains a sacrificial reaction substance 321, and the electrode activation method provided in this embodiment requires the participation of the sacrificial reactant 321 to react (this reaction can be a pure electrochemical reaction or doped with other chemical reactions, preferably pure Electrochemical reaction), this reaction can partially activate the working electrode and the counter electrode, and does not change the hydrophilic and hydrophobic properties of the inert region 24, does not produce chemical reactions with the inert region 24, and at the same time, the sacrificial reactant 321 Contamination of the inert area 24 due to production problems can be removed to a certain extent.
  • the sacrificial reactants 321 include one or more of acids, bases, oxides or reducing substances, but are certainly not limited thereto, and can be selected and adjusted according to actual needs or possible needs.
  • the molar concentration of the aqueous solution of the sacrificial reactant 321 ranges from 0.1 mM to 500 mM.
  • the molar concentration of the aqueous solution of the sacrificial reactant 321 may range from 100 mM to 500 mM.
  • the sacrificial reactant 321 includes potassium salts, such as potassium ferricyanide and potassium chloride.
  • Step 103 controlling the signal generator to apply an activation electrical signal to the biosensor chip, so as to activate the working electrode and/or the counter electrode through the electrolyte.
  • control signal generator 41 applies an activation electrical signal to the biosensor chip to activate the working electrode 22 and/or the counter electrode 21 through the electrolyte 32, wherein the potential of the activation electrical signal is lower than the electric potential of the electrical activation signal. Oxidation potential of catalytic species 23.
  • the activation electrical signal may include a DC voltage signal, an AC voltage signal, or a DC voltage signal superimposed with an AC voltage signal.
  • the DC voltage signal when the activation electrical signal is a DC voltage signal, the DC voltage signal includes a stepped DC voltage signal.
  • the scanning rate (scan rate) of the stepped DC voltage signal ranges from 0.001V/s to 1.0V/s, and of course it is not limited thereto, and can be selected and adjusted according to actual or possible needs.
  • the activation electrical signal when the activation electrical signal is an AC voltage signal or a DC voltage signal superimposed with an AC voltage signal, the AC voltage signal may include any of the sine wave, triangular wave, sawtooth wave, square wave, and preset wave AC voltage signal.
  • One or more types are not limited thereto, and can be selected and adjusted according to actual needs or possible needs.
  • the frequency range of the AC voltage signal may be 0.1 Hz-1 GHz, preferably, 1 Hz-1 KHz.
  • the direction of the activation electrical signal may include any one or more of unidirectional oxidation, unidirectional reduction, and bidirectional symmetrical redox.
  • the potential range of the activation electrical signal is -1.2V to 1.8V, preferably, it can be -0.8V to 1.6V. Select and adjust accordingly.
  • the total time period for applying the activation electrical signal ranges from 1 to 100 activation reaction cycles, preferably, it can be 10 to 50 activation reaction cycles, and of course it is not limited thereto. Make corresponding selections and adjustments based on actual needs or possible needs.
  • the increase range of the active area after activation is 1-1000 times.
  • FIG. 3A to FIG. 3F respectively show six types of signal schematic diagrams of activation electrical signals applied by the signal generator, that is, the following six types of signals respectively.
  • the signal generator is used to apply a stepped DC voltage signal
  • the scanning speed of the electrical signal is 0.1V/s
  • the scanning potential window is 0-1.6V
  • the total time period of applying the activation electrical signal is 10 activation reaction cycles
  • the scanning direction is bidirectional symmetrical redox.
  • the scanning speed of the electrical signal is 0.1V/s
  • the scanning potential window is 0-1.6V
  • the total time period for applying the activation electrical signal is 10 activation reaction cycles
  • the scanning direction is bidirectional symmetrical redox.
  • the signal generator is used to apply a stepped DC voltage signal, the scanning speed of the electrical signal is 0.1V/s, the scanning potential window is 0-1.6V, and the total time period of applying the activation electrical signal It is 10 activation reaction cycles, and the scanning direction is unidirectional oxidation.
  • the fourth type of signal as shown in Figure 3D, use a signal generator to apply a triangular wave AC voltage signal, the scanning speed of the electrical signal is 0.1V/s, the scanning potential window is 0-1.6V, and the total time period for applying the activation electrical signal It is 10 activation reaction cycles, and the scanning direction is unidirectional oxidation.
  • a signal generator is used to apply a stepped DC voltage signal, the scanning speed of the electrical signal is 0.1V/s, the scanning potential window is 0-1.6V, and the total time period of applying the activation electrical signal It is 10 activation reaction cycles, and the scanning direction is unidirectional reduction.
  • the sixth type of signal as shown in Figure 3F, use a signal generator to apply a triangular wave AC voltage signal, the scanning speed of the electrical signal is 0.1V/s, the scanning potential window is 0-1.6V, and the total time period for applying the activation electrical signal It is 10 activation reaction cycles, and the scanning direction is unidirectional reduction.
  • the preparation of the sacrificial reactant activation solution is 100 millimoles per liter (mM) of NaOH, 1% H 2 O 2 and 150mM NaCl, uniformly mixed before use; the working electrode is Ti, and the electrocatalytic substance is a noble metal platinum layer, passed Physical vapor deposition electron beam evaporation is bonded to the working electrode with an inert area of SU-8.
  • Fig. 4 is a schematic diagram showing a comparison of the contact angles of the inert regions before and after activation by the electrode activation method of the biosensing chip. Referring to FIG. 4 , the contact angles of water in the inert region before and after activation did not change significantly, being 121.413° and 121.096° respectively, both of which are hydrophobic.
  • FIG. 5 is a schematic diagram showing the comparison of scan rate images before and after activation by the electrode activation method of the biosensing chip.
  • I p (2.69 ⁇ 10 5 )n 3/2 AD 1/2 C* ⁇ 1/2
  • y 3.4625 ⁇ 10 -7 x+7 ⁇ 10 - 8
  • R 2 0.9986, where y represents Ip and x represents ⁇ 1/2 . It can be calculated that the effective area after activation is 312% of the effective area before activation, greatly increasing the effective area.
  • Table 1 shows the parameter statistics of the influence of the scan rate on the cyclic voltammetry curve after electroactivation.
  • Fig. 6 is a schematic diagram showing the comparison of multi-sweep cyclic voltammetry before and after activation by the electrode activation method of the biosensing chip.
  • the multi-sweep cyclic voltammetry after activation is excellent, the peak potential difference is less than 120mV, the peak current ratio is close to 1, and the reversibility is strong.
  • the peak current and peak potential change range is almost No change, strong corrosion resistance; before activation, the cyclic voltammetry is not good, the peak potential difference is close to 300mV, the redox current becomes lower and lower with the increase of the number of scans, and gradually becomes 0, poor reversibility, corrosion resistance Sex is poor.
  • Fig. 7 is a schematic diagram showing the comparison of open circuit potentials before and after activation by the electrode activation method of the biosensing chip.
  • the open circuit potential time curve technique was used to characterize the electrode activation effect, and the characterization solution used 5mM potassium ferricyanide, 5mM potassium ferrocyanide plus 0.1M KCl.
  • the open circuit potentials before and after activation were 0.234V and 0.231V respectively, and there was no significant difference in the internal potential of the electrodes before and after electroactivation treatment.
  • Fig. 8 shows the AC impedance Nyquist (Nyquist frequency) figure before and after activation of the electrode activation method of the biosensor chip, and the potassium ferrocyanide of 5mM, the potassium ferrocyanide of 5mM adds 0.1M KCl.
  • R s represents the solution resistance
  • R represents the internal resistance of the biosensing chip
  • R ct represents the charge transfer resistance
  • the corresponding value can be obtained by importing the original data of the measured AC impedance into the Zview software. Calculation shows that the charge transfer resistance after activation drops to 2.9% of that before activation.
  • FIG. 9 is a three-dimensional schematic diagram showing the impedance before and after fitting by the electrode activation method of the biosensing chip
  • FIG. 10 is a schematic diagram showing the equivalent circuit fitting of the electrode activation method of the biosensing chip.
  • the fitted circuit after activation is a standard Randles circuit.
  • the electrode activation method of the biosensing chip provided in this embodiment plays a key role in improving the performance of the counter electrode and the working electrode, especially for the manufacturing process of the high-end chip of the second and third types of medical equipment or the pretreatment process before use.
  • Reference function: The electrode activation method of the biosensor chip provided in this embodiment has a simple process flow, and the activation solution of the sacrificial reactant is easy to prepare, clean and harmless, and the type and potential range of the activation electrical signal applied by the signal generator can be artificially Controlling, and simultaneously activating the counter electrode and the working electrode, reduces the time cost caused by separate operations, locally activates the sensing area while ensuring the hydrophilic and hydrophobic properties of the inert area, and ensures the stability of the modified substrate for other processes.
  • This embodiment also provides a biosensing chip, which uses the method for activating electrodes of the biosensing chip as in the above embodiment to activate the electrodes.
  • This embodiment also provides an electrode activation device for a biosensing chip, which is used to implement the electrode activation method of the biosensing chip as in the above embodiment, and is used for the biosensing chip as in the above embodiment Electrode activation is performed, so the structure of the biosensing chip can refer to the structure of the biosensing chip in the above embodiment.
  • the electrode activation device mainly includes a container and a signal generator.
  • the container is used to hold the electrolyte, and when the electrode is activated, the biosensing chip is placed in the container containing the electrolyte, so that the working electrode and the counter electrode are respectively immersed in the electrolyte;
  • the signal generator is configured as An activation electrical signal is applied to the biosensing chip to activate the working electrode and/or the counter electrode through the electrolyte, wherein the potential of the activation electrical signal is lower than the oxidation potential of the electrocatalytic substance.
  • the biosensing chip and its electrode activation method and device provided in this embodiment enhance the electrochemical performance of the micro-nano biosensing chip, greatly reduce the charge transfer resistance and increase the active effective area, and can control the chip between batches/batches. Internal electrochemical differences can reduce the coefficient of variation of performance, which can effectively ensure the consistent electrochemical quality of the chip before use, and can also partially treat the reaction area without affecting the hydrophilic and hydrophobic properties of the inert area, especially for the pre-treatment of high-end chips for Class II and Class III medical devices.
  • the processing usage specification provides a reference, which is beneficial to the research and development of biosensing chips.

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Abstract

一种生物传感芯片及其电极活化方法和装置，生物传感芯片包括载板、工作电极（22）及对电极（21），载板上分别形成有惰性区域（24）及工作电极（22），工作电极（22）具有电催化物质；电极活化方法包括：将生物传感芯片放置于容纳有电解液（32）的容器（31）中，以使工作电极（22）及对电极（21）分别浸泡在电解液（32）中；控制信号发生器向生物传感芯片施加活化电信号，以通过电解液（32）活化工作电极（22）和/或对电极（21），其中活化电信号的电位小于电催化物质的氧化电位。本申请增强了微纳生物传感芯片的电化学性能，大幅降低了电荷转移电阻并提高了活性有效面积，能控制芯片批间/批内电化学差异，减小性能变异系数，使用前能有效地保证芯片电化学性能一致同时能局部处理反应区域，不影响惰性区域的亲疏水性质。

Description

生物传感芯片及其电极活化方法和装置 技术领域
本申请涉及电化学技术领域，特别涉及一种生物传感芯片及其电极活化方法和装置。
背景技术
电化学学科分支分为电池(如锂电池)、超级电容器和生物传感，其中电化学生物传感在医疗器械中充当着重要角色。现如今医疗器械，尤其是体外诊断(IVD)领域，根据检测项目的不同，多种方法学共同竞争市场占有率。主流的平台有化学发光(电化学发光)、特定蛋白、生化、免疫、血凝、血球、血气、胶体金、分子诊断、POCT(point of care testing，即时检验)等，电化学生物传感涉及的项目包含定频交流阻抗血凝芯片、电流/电压/电导法干式血气生化芯片/湿式电极、一二三代生物传感器血糖试纸/三类医疗器械植入性持续监测血糖电极和高端基因测序-纳米孔道电化学单分子基因测序芯片等。
电化学生物传感所采用的基材芯片一般有丝网印刷电极(SPE)、印刷电路板(PCB)、柔性电路板(FPCB)和半导体(Semi-conductor)芯片；所采用的技术工艺有丝网印刷工艺、PCB工艺和MEMS(微机电系统)工艺。由于生物传感芯片对芯片生产工艺敏感度较高，生产过程中残留的浆料、光刻胶、阻焊漆、镀层药水结晶沉淀等其他杂质，因工艺中对传感区域的物理/化学清洗不到位，曝光、显影、蚀刻欠佳或储存过程中的金属钝化氧化失效会对整个电化学反应乃至电信号采集产生巨大影响，直接影响到电化学性能的电荷转移电阻和活性有效面积，甚至是生物传感芯片使用寿命。所以无论是采用任何一种基材芯片，都需要对芯片电极传感区域进行清洁预处理和活化处理，提高电化学反应信号采集有效性和准确性。
由于电化学生物传感芯片除了电极活性区域-反应区域最为重要外，维持保护引线或反应槽的惰性区域的亲疏水性也尤为重要。量产工艺的等离子体(如氧气、氩气、氦气和氮气等)对芯片全方位羟基、羧基或氨基活化处理后，会导致芯片惰性区域同样改性。同样地，使用硅烷化疏水性质复性，也会导致芯片电极活性区域同样改性，直接引起导通不良。
综上所述，目前，矩阵电极或高端生物传感芯片制造工艺，做到电极集成度越高、面积越小如微纳加工技术，在应用到电化学生物传感产品项目开发阶段极大程度会出现电学不导通或性能不佳等产品致命缺陷，简单的物理抛光、切割、清洗冲刷或者化学浸泡均不能有效地解决问题，过度的物理化学轰击，如等离子体清洗，并不能理想地局部处 理金属部位，干扰产品开发的其他部位修饰步骤；同时，溯源到生产工艺上并改进工艺确实能一定程度解决上述电学惰性物质残留问题，但不能有效保证生产批间/批内差、运输不确定度、存储不确定度和稳定性不确定度等的存在不影响产品电化学性能的一致性。
发明内容
本申请的主要目的在于，提供一种生物传感芯片及其电极活化方法和装置，以改善现有技术中存在的上述缺陷。
本申请是通过下述技术方案来解决上述技术问题：
作为本申请的第一方面，提供一种生物传感芯片的电极活化方法，所述生物传感芯片包括载板、工作电极及对电极，所述载板上分别形成有惰性区域及所述工作电极，所述工作电极具有电催化物质；
所述电极活化方法包括以下步骤：
将所述生物传感芯片放置于容纳有电解液的容器，以使所述工作电极及所述对电极分别浸泡在所述电解液中；
控制信号发生器向所述生物传感芯片施加活化电信号，以通过所述电解液活化所述工作电极和/或所述对电极，其中所述活化电信号的电位小于所述电催化物质的氧化电位。
作为可选实施方式，所述工作电极包括至少一个工作电极单元。
作为可选实施方式，所述工作电极包括至少两个呈周期性阵列分布的所述工作电极单元。
作为可选实施方式，所述对电极与所述工作电极单元的数量比范围为1:16～1:4096。
作为可选实施方式，每一个所述工作电极单元分别具有电催化物质。
作为可选实施方式，所述电催化物质包括石墨烯、贵金属、胶体金或前述物质的复合物中的任意一种或多种。
作为可选实施方式，在控制信号发生器向所述生物传感芯片施加活化电信号的步骤中，
所述活化电信号包括直流电压信号、交流电压信号或叠有交流电压信号的直流电压信号。
作为可选实施方式，当所述活化电信号包括直流电压信号时，所述直流电压信号包括阶梯直流电压信号。
作为可选实施方式，所述阶梯直流电压信号的扫描速度范围为0.001V/s～1.0V/s。
作为可选实施方式，当所述活化电信号包括交流电压信号或叠有交流电压信号的直 流电压信号时，所述交流电压信号包括正弦波、三角波、锯齿波、方波及预设波交流电压信号中的任意一种或多种。
作为可选实施方式，当所述活化电信号包括交流电压信号或叠有交流电压信号的直流电压信号时，所述交流电压信号的频率为0.1Hz～1GHz。
作为可选实施方式，所述交流电压信号的频率为1Hz～1KHz。
作为可选实施方式，所述活化电信号的方向包括单方向氧化、单方向还原及双方向对称氧化还原中的任意一种或多种。
作为可选实施方式，所述活化电信号的电位范围为-1.2V～1.8V。
作为可选实施方式，在控制信号发生器向所述生物传感芯片施加活化电信号的步骤中，施加活化电信号的总时间段范围为1～100个活化反应周期。
作为可选实施方式，施加活化电信号的总时间段范围为10～50个活化反应周期。
作为可选实施方式，所述电解液包含牺牲反应物质，所述牺牲反应物包括酸、碱、氧化物或还原性物质中的一种或多种。
作为可选实施方式，所述牺牲反应物的水溶液的摩尔浓度范围为0.1mM～500mM。
作为可选实施方式，所述惰性区域中的惰性物质包括SU-8(光刻胶)、环氧树脂、聚酰亚胺、FR4(耐燃材料等级的代号)板及玻璃纤维层中的任意一种或多种。
作为本申请的第二方面，提供一种生物传感芯片，利用如上述的生物传感芯片的电极活化方法来进行电极活化。
作为本申请的第三方面，提供一种生物传感芯片的电极活化装置，所述生物传感芯片包括载板、工作电极及对电极，所述载板上分别形成有惰性区域及所述工作电极，所述工作电极具有电催化物质；
所述电极活化装置包括：
容器，用于容纳电解液，在进行电极活化时，将所述生物传感芯片放置于容纳有所述电解液的所述容器中，以使所述工作电极及所述对电极分别浸泡在所述电解液中；以及，
信号发生器，被配置为向所述生物传感芯片施加活化电信号，以通过所述电解液活化所述工作电极和/或所述对电极，其中所述活化电信号的电位小于所述电催化物质的氧化电位。
根据本申请内容，本领域技术人员可以理解本申请内容的其它方面。
本申请的积极进步效果在于：
本申请提供的生物传感芯片及其电极活化方法和装置，增强了微纳生物传感芯片的 电化学性能，大幅降低了电荷转移电阻并提高了活性有效面积，能控制芯片批间/批内电化学差异，减小性能变异系数，使用前能有效地保证芯片电化学素质一致同时能局部处理反应区域，不影响惰性区域的亲疏水性质，尤其为二类三类医疗器械高端芯片的预处理使用规范提供了参考，有利于生物传感芯片的研发。
附图说明
在结合以下附图阅读本申请的实施例的详细描述之后，能够更好地理解本申请的所述特征和优点。在附图中，各组件不一定是按比例绘制，并且具有类似的相关特性或特征的组件可能具有相同或相近的附图标记。
图1为根据本申请的示例性实施例的生物传感芯片的电极活化方法的流程示意图。
图2为根据本申请的示例性实施例的生物传感芯片及其电极活化装置的部分结构示意图。
图3A为根据本申请的示例性实施例的信号发生器施加的活化电信号的第一类型信号示意图。
图3B为根据本申请的示例性实施例的信号发生器施加的活化电信号的第二类型信号示意图。
图3C为根据本申请的示例性实施例的信号发生器施加的活化电信号的第三类型信号示意图。
图3D为根据本申请的示例性实施例的信号发生器施加的活化电信号的第四类型信号示意图。
图3E为根据本申请的示例性实施例的信号发生器施加的活化电信号的第五类型信号示意图。
图3F为根据本申请的示例性实施例的信号发生器施加的活化电信号的第六类型信号示意图。
图4为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的惰性区域接触角对比示意图。
图5为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的扫描速度影像对比示意图。
图6为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的多扫循环伏安对比示意图。
图7为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的 开路电位对比示意图。
图8为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的交流阻抗对比示意图。
图9为根据本申请的示例性实施例的生物传感芯片的电极活化方法进行活化前后的阻抗三维示意图。
图10为根据本申请的示例性实施例的生物传感芯片的电极活化方法的等效电路拟合示意图。
具体实施方式
下面通过实施例的方式进一步说明本申请，但并不因此将本申请限制在所述的实施例范围之中。
应当注意，在说明书中对“一实施例”、“可选实施例”、“另一实施例”等的引用指示所描述的实施例可以包括特定的特征、结构或特性，但是每个实施例可能不一定包括该特定的特征、结构或特性。而且，这样的短语不一定指代相同的实施例。此外，当结合实施例描述特定特征、结构或特性时，无论是否被明确描述，结合其它实施例来实现这样的特征、结构或特性都在相关领域的技术人员的知识范围内。
在本申请内容的描述中，需要理解的是，术语“中心”、“横向”、“上”、“下”、“左”、“右”、“竖直”、“水平”、“顶”、“底”、“内”、“外”等指示的方位或位置关系为基于附图所示的方位或位置关系，仅是为了便于描述本申请内容和简化描述，而不是指示或暗示所指的装置或元件必须具有特定的方位、以特定的方位构造和操作，因此不能理解为对本申请内容的限制。此外，术语“第一”、“第二”仅用于描述目的，而不能理解为指示或暗示相对重要性或者隐含指明所指示的技术特征的数量。由此，限定有“第一”、“第二”的特征可以明示或者隐含地包括一个或者更多个该特征。在本申请内容的描述中，除非另有说明，“多个”的含义是两个或两个以上。另外，术语“包括”及其任何变形，意图在于覆盖不排他的包含。
在本申请内容的描述中，需要说明的是，除非另有明确的规定和限定，术语“安装”、“相连”、“连接”应做广义理解，例如，可以是固定连接，也可以是可拆卸连接，或一体地连接；可以是机械连接，也可以是电连接；可以是直接相连，也可以通过中间媒介间接相连，可以是两个元件内部的连通。对于本领域的普通技术人员而言，可以具体情况理解上述术语在本申请内容中的具体含义。
这里所使用的术语仅仅是为了描述具体实施例而不意图限制示例性实施例。除非上 下文明确地另有所指，否则这里所使用的单数形式“一个”、“一项”还意图包括复数。还应当理解的是，这里所使用的术语“包括”和/或“包含”规定所陈述的特征、整数、步骤、操作、单元和/或组件的存在，而不排除存在或添加一个或更多其他特征、整数、步骤、操作、单元、组件和/或其组合。
为了克服目前存在的上述缺陷，本实施例提供一种生物传感芯片的电极活化方法，生物传感芯片包括载板、工作电极及对电极，载板上分别形成有惰性区域及工作电极，工作电极具有电催化物质；电极活化方法包括以下步骤：将生物传感芯片放置于容纳有电解液的容器，以使工作电极及对电极分别浸泡在电解液中；控制信号发生器向生物传感芯片施加活化电信号，以通过电解液活化工作电极和/或对电极，其中活化电信号的电位小于电催化物质的氧化电位。
在本实施例中，该电极活化方法除了优选应用于生物传感芯片领域之外，还可应用于半导体、高端制造业、其他医疗器械等其他领域，本实施例并不具体限定该电极活化方法的应用领域，可根据实际需求或可能出现的需求进行相应的选择及调整。
在本实施例中，生物传感芯片的电极采用1对n(n为大于或等于1的自然数)的电流回路二电极体系，无需外接参比电极，其中，对电极可以为宏电极，工作电极可以为微矩阵电极，但并不具体限定电极类型，可根据实际需求或可能出现的需求进行相应的选择及调整。
在本实施例中，利用该电极活化方法，电催化物质在工作电极和/或对电极的电活化过程中不被消耗或剥离，同时能够大幅增加对电极与工作电极活性面积；该电极活化方法通过采用电动势驱动力(EMF)的信号发生器向生物传感芯片施加活化电信号，而且施加电位区间不会钝化宏/微矩阵电极的活性反应层，但会氧化或还原电极表面惰性物质，活化的同时起到加速清洁作用。
具体地，作为一较佳实施例，如图1所示，本实施例提供的生物传感芯片的电极活化方法主要包括以下步骤：
步骤101、提供待活化的生物传感芯片。
作为一具体实施方式，以下说明本步骤中提供的生物传感芯片，但该生物传感芯片应当并不仅限于如下说明的结构，可根据实际需求或可能出现的需求进行相应的选择及调整。
参考图2所示，生物传感芯片主要包括载板(图中未示出)、对电极21及工作电极22，载板上分别形成有惰性区域24及工作电极22，工作电极22具有电催化物质23。具体地，工作电极22表面负载电催化物质。
其中，对电极21可以为生物传感芯片的宏电极，工作电极22可以包括至少一个工作电极单元221。
作为一可选实施方式，工作电极22可以通过至少两个呈周期性阵列分布的工作电极单元221来形成，即工作电极22可以为微矩阵电极，但并不具体限定工作电极单元221的数量及分布形状，均可根据实际需求或可能出现的需求进行相应的选择及调整。
作为较佳实施方式，对电极21与工作电极单元221的数量比范围为1:16～1:4096，即当生物传感芯片包括一个对电极21时，生物传感芯片可以包括16个～4096个工作电极单元221，当然并不仅限于此数量比范围。优选地，对电极21与工作电极单元221的数量比为1:256。
具体地，每一个工作电极单元221分别具有对应的电催化物质23。
作为较佳实施方式，电催化物质23可以包括石墨烯、贵金属、胶体金或前述物质的复合物中的任意一种或多种，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
进一步地，石墨烯可以为三维石墨烯，金可以为粗糙化后的金，贵金属可以为贵金属层或颗粒，贵金属具体可以为铂或钯。
作为较佳实施方式，电催化物质23可通过电化学沉积、化学沉积、喷阀点胶及物理气相沉积中的任意一种或多种方式结合至工作电极单元221，当然并不仅限于此。优选地，物理气相沉积可以为磁控溅射。
在本实施例中，电催化物质能降低电解反应理论所需电压值，降低过电位。
作为具体实施方式，惰性区域24中的惰性物质可以包括SU-8、环氧树脂、聚酰亚胺、FR4板及玻璃纤维层中的任意一种或多种，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。优选地，惰性物质可以为SU-8。
步骤102、将生物传感芯片放置于容纳有电解液的容器，以使工作电极及对电极分别浸泡在电解液中。
参考图2所示，将步骤101中提供的生物传感芯片放置于容纳有电解液32的容器31，以使每一个工作电极单元221及对电极21分别浸泡在电解液32中。
作为具体实施方式，电解液32包含牺牲反应物质321，本实施例提供的电极活化方法需要牺牲反应物321的参与进而反应(该反应可以为纯电化学反应或掺杂其他化学反应，优选为纯电化学反应)，该反应能局部活化工作电极及对电极，而且不改变惰性区域24的亲疏水性质，不与惰性区域24产生化学反应，同时在不施加电极活化方法下，该牺牲反应物321能一定程度去除由于生产问题导致的惰性区域24的脏污。
牺牲反应物321包括酸、碱、氧化物或还原性物质中的一种或多种，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
作为较佳实施方式，牺牲反应物321的水溶液的摩尔浓度范围为0.1mM～500mM。优选地，牺牲反应物321的水溶液的摩尔浓度范围可以为100mM～500mM。
作为一可选实施方式，牺牲反应物321包括钾盐，例如铁氰化钾和氯化钾等。
步骤103、控制信号发生器向生物传感芯片施加活化电信号，以通过电解液活化工作电极和/或对电极。
在本步骤中，参考图2所示，控制信号发生器41向生物传感芯片施加活化电信号，以通过电解液32活化工作电极22和/或对电极21，其中活化电信号的电位小于电催化物质23的氧化电位。
具体地，在本步骤中，活化电信号可以包括直流电压信号、交流电压信号或叠有交流电压信号的直流电压信号。
作为可选实施方式，当活化电信号为直流电压信号时，直流电压信号包括阶梯直流电压信号。
具体地，阶梯直流电压信号的扫描速度(scan rate)范围为0.001V/s～1.0V/s，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
作为可选实施方式，当活化电信号为交流电压信号或叠有交流电压信号的直流电压信号时，交流电压信号可以包括正弦波、三角波、锯齿波、方波及预设波交流电压信号中的任意一种或多种，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
具体地，交流电压信号的频率范围可以为0.1Hz～1GHz，优选地，可以为1Hz～1KHz。
在本步骤中，活化电信号的方向可以包括单方向氧化、单方向还原及双方向对称氧化还原中的任意一种或多种。
作为可选实施方式，在本步骤中，活化电信号的电位范围为-1.2V～1.8V，优选地，可以为-0.8V～1.6V，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
作为可选实施方式，在本步骤中，施加活化电信号的总时间段范围为1～100个活化反应周期，优选地，可以为10～50个活化反应周期，当然并不仅限于此，可根据实际需求或可能出现的需求进行相应的选择及调整。
在本实施例中，进行活化后活性面积的增加幅度为1～1000倍。
以下作为具体较佳实施方式，结合附图所示提供如上述实施例的生物传感芯片的电 极活化方法的具体应用场景和活化前后的效果。
图3A至图3F分别示出信号发生器施加的活化电信号的六种类型信号示意图，即分别为以下六种类型信号。
作为第一类型信号，参考图3A所示，采用信号发生器施加阶梯直流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为双方向对称氧化还原。
作为第二类型信号，参考图3B所示，采用信号发生器施加三角波交流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为双方向对称氧化还原。
作为第三类型信号，参考图3C所示，采用信号发生器施加阶梯直流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为单方向氧化。
作为第四类型信号，参考图3D所示，采用信号发生器施加三角波交流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为单方向氧化。
作为第五类型信号，参考图3E所示，采用信号发生器施加阶梯直流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为单方向还原。
作为第六类型信号，参考图3F所示，采用信号发生器施加三角波交流电压信号，电信号的扫描速度为0.1V/s，扫描电位窗口在0～1.6V，施加活化电信号的总时间段为10个活化反应周期，扫描方向为单方向还原。
牺牲反应物活化液的配制为100毫摩尔每升(mM)的NaOH、1％的H 2O 2和150mM的NaCl，使用前均匀混合；工作电极为Ti，电催化物质为贵金属铂层，通过物理气相沉积电子束蒸发结合到工作电极上，惰性区域为SU-8。
图4示出生物传感芯片的电极活化方法进行活化前后的惰性区域接触角对比示意图。参考图4所示，进行活化前后的惰性区域的水的接触角无明显变化，分别为121.413°和121.096°，皆为疏水性质。
参考图5-6采用循环伏安法表征电极活化效果，表征液使用10mM的铁氰化钾加0.1M的KCl。
图5为示出了生物传感芯片的电极活化方法进行活化前后的扫描速率影像对比示意图。参考图5所示，进行活化前后不同扫描速率对循环伏安曲线的影响，根据Randles- Sevcik公式I p＝(2.69×10 5)n 3/2AD 1/2C*ν 1/2，得到电活化后线性方程为y＝3.4625×10 -7x+7×10 - 8,R 2＝0.9986，式中，y代表Ip，x代表ν 1/2。可计算得到活化后有效面积为活化前有效面积的312％，大幅增加了有效面积。
如下表1为电活化后的扫描速率对循环伏安曲线的影响的参数统计。
表1：
Figure PCTCN2021141710-appb-000001
Figure PCTCN2021141710-appb-000002
图6示出生物传感芯片的电极活化方法进行活化前后的多扫循环伏安对比示意图。参考图6所示，进行活化后多扫循环伏安表征优秀，峰电位差均小于120mV，峰电流比值都接近1，可逆性强，随着扫描次数的增加，峰电流及峰电位变化幅度几乎不变，耐腐蚀性强；进行活化前，循环伏安表征不佳，峰电位差均接近300mV，氧化还原电流随着扫描次数增加越来越低，并逐渐变为0，可逆性差，耐腐蚀性较差。
图7示出生物传感芯片的电极活化方法进行活化前后的开路电位对比示意图。采用开路电位时间曲线技术表征电极活化效果，表征液使用5mM的铁氰化钾、5mM的亚铁氰化钾加0.1M的KCl。参考图7所示，进行活化前后的开路电位分别为0.234V和0.231V，通过电活化处理前后电极的内电势并未出现明显差异。
图8为示出生物传感芯片的电极活化方法进行活化前后的交流阻抗Nyquist(奈奎斯特频率)图，表征液使用5mM的铁氰化钾、5mM的亚铁氰化钾加0.1M的KCl。参考图8所示，半圆弧直径对比，活化后阻抗明显降低；进行活化前R s+R 内＝7400Ω,R ct＝3492177Ω；进行活化后R s+R 内＝6636Ω,R ct＝100720Ω，其中R s代表溶液电阻，R 内代表生物传感芯片内阻；Rct为电荷转移电阻，相应值可通过将测得的交流阻抗原始数据导入到Zview软件得到。计算可知电荷转移电阻活化后下降至活化前的2.9％。
图9为示出生物传感芯片的电极活化方法进行活化后拟合前后的阻抗三维示意图，图10为示出生物传感芯片的电极活化方法的等效电路拟合示意图。参考图9及图10所示，等效电路拟合各参数分别为R s＝6636,CPE dl-T＝4.855×10 9,CPE dl-P＝0.81874,R ct＝100720, W o1-R＝333050,W o1-T＝0.12735,W o1-P＝0.5，其中CPE代表恒相位阻抗，W代表扩散阻抗，上述数值可通过Zview软件得到。活化后拟合电路为标准Randles(兰德尔斯)电路。
本实施例提供的生物传感芯片的电极活化方法，针对对电极和工作电极的性能提升起到关键作用，尤其对二类、三类医疗器械高端芯片的制造工艺或使用前预处理工艺具有明显借鉴作用；本实施例提供的生物传感芯片的电极活化方法，其工艺流程简单，牺牲反应物的活化液配制简易、清洁无害，信号发生器施加的活化电信号发生种类及电位区间可人为调控，而且将对电极及工作电极同时活化，减少了单独操作所带来的时间成本，局部活化传感区域的同时保证了惰性区域的亲疏水性质，为其他工艺保证了修饰基底的稳定性。
本实施例还提供一种生物传感芯片，利用如上述实施例的生物传感芯片的电极活化方法来进行电极活化。
本实施例还提供一种生物传感芯片的电极活化装置，该电极活化装置用于实现如上述实施例的生物传感芯片的电极活化方法，并且用于针对如上述实施例的生物传感芯片进行电极活化，因此生物传感芯片的结构可参考如上述实施例的生物传感芯片的结构。
该电极活化装置主要包括容器及信号发生器。其中，容器用于容纳电解液，在进行电极活化时，将生物传感芯片放置于容纳有电解液的容器中，以使工作电极及对电极分别浸泡在电解液中；信号发生器被配置为向生物传感芯片施加活化电信号，以通过电解液活化工作电极和/或对电极，其中活化电信号的电位小于电催化物质的氧化电位。
本实施例提供的生物传感芯片及其电极活化方法和装置，增强了微纳生物传感芯片的电化学性能，大幅降低了电荷转移电阻并提高了活性有效面积，能控制芯片批间/批内电化学差异，减小性能变异系数，使用前能有效地保证芯片电化学素质一致同时能局部处理反应区域，不影响惰性区域的亲疏水性质，尤其为二类三类医疗器械高端芯片的预处理使用规范提供了参考，有利于生物传感芯片的研发。
虽然以上描述了本申请的具体实施方式，但是本领域的技术人员应当理解，这仅是举例说明，本申请的保护范围是由所附权利要求书限定的。本领域的技术人员在不背离本申请的原理和实质的前提下，可以对这些实施方式做出多种变更或修改，但这些变更和修改均落入本申请的保护范围。

Claims (21)

  1. 一种生物传感芯片的电极活化方法，其特征在于，所述生物传感芯片包括载板、工作电极及对电极，所述载板上分别形成有惰性区域及所述工作电极，所述工作电极具有电催化物质；
    所述电极活化方法包括以下步骤：
    将所述生物传感芯片放置于容纳有电解液的容器，以使所述工作电极及所述对电极分别浸泡在所述电解液中；
    控制信号发生器向所述生物传感芯片施加活化电信号，以通过所述电解液活化所述工作电极和/或所述对电极，其中所述活化电信号的电位小于所述电催化物质的氧化电位。
  2. 如权利要求1所述的电极活化方法，其特征在于，所述工作电极包括至少一个工作电极单元。
  3. 如权利要求2所述的电极活化方法，其特征在于，所述工作电极包括至少两个呈周期性阵列分布的所述工作电极单元。
  4. 如权利要求2所述的电极活化方法，其特征在于，所述对电极与所述工作电极单元的数量比范围为1:16～1:4096。
  5. 如权利要求2所述的电极活化方法，其特征在于，每一个所述工作电极单元分别具有电催化物质。
  6. 如权利要求1所述的电极活化方法，其特征在于，所述电催化物质包括石墨烯、贵金属、胶体金或前述物质的复合物中的任意一种或多种。
  7. 如权利要求1所述的电极活化方法，其特征在于，在控制信号发生器向所述生物传感芯片施加活化电信号的步骤中，
    所述活化电信号包括直流电压信号、交流电压信号或叠有交流电压信号的直流电压信号。
  8. 如权利要求7所述的电极活化方法，其特征在于，当所述活化电信号包括直流电压信号时，所述直流电压信号包括阶梯直流电压信号。
  9. 如权利要求8所述的电极活化方法，其特征在于，所述阶梯直流电压信号的扫描速度范围为0.001V/s～1.0V/s。
  10. 如权利要求7所述的电极活化方法，其特征在于，当所述活化电信号包括交流电压信号或叠有交流电压信号的直流电压信号时，所述交流电压信号包括正弦波、三角波、锯齿波、方波及预设波交流电压信号中的任意一种或多种。
  11. 如权利要求7所述的电极活化方法，其特征在于，当所述活化电信号包括交流电压信号或叠有交流电压信号的直流电压信号时，所述交流电压信号的频率为0.1Hz～1GHz。
  12. 如权利要求11所述的电极活化方法，其特征在于，所述交流电压信号的频率为1Hz～1KHz。
  13. 如权利要求7所述的电极活化方法，其特征在于，所述活化电信号的方向包括单方向氧化、单方向还原及双方向对称氧化还原中的任意一种或多种。
  14. 如权利要求7所述的电极活化方法，其特征在于，所述活化电信号的电位范围为-1.2V～1.8V。
  15. 如权利要求1所述的电极活化方法，其特征在于，在控制信号发生器向所述生物传感芯片施加活化电信号的步骤中，施加活化电信号的总时间段范围为1～100个活化反应周期。
  16. 如权利要求15所述的电极活化方法，其特征在于，施加活化电信号的总时间段范围为10～50个活化反应周期。
  17. 如权利要求1所述的电极活化方法，其特征在于，所述电解液包含牺牲反应物质，所述牺牲反应物包括酸、碱、氧化物或还原性物质中的一种或多种。
  18. 如权利要求17所述的电极活化方法，其特征在于，所述牺牲反应物的水溶液的摩尔浓度范围为0.1mM～500mM。
  19. 如权利要求1所述的电极活化方法，其特征在于，所述惰性区域中的惰性物质包括SU-8、环氧树脂、聚酰亚胺、FR4板及玻璃纤维层中的任意一种或多种。
  20. 一种生物传感芯片，其特征在于，利用如权利要求1～19中任意一项所述的生物传感芯片的电极活化方法来进行电极活化。
  21. 一种生物传感芯片的电极活化装置，其特征在于，所述生物传感芯片包括载板、工作电极及对电极，所述载板上分别形成有惰性区域及所述工作电极，所述工作电极具有电催化物质；
    所述电极活化装置包括：
    容器，用于容纳电解液，在进行电极活化时，将所述生物传感芯片放置于容纳有所述电解液的所述容器中，以使所述工作电极及所述对电极分别浸泡在所述电解液中；以及，
    信号发生器，被配置为向所述生物传感芯片施加活化电信号，以通过所述电解液活化所述工作电极和/或所述对电极，其中所述活化电信号的电位小于所述电催化物质的氧化电位。
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CN113567527A (zh) * 2021-06-02 2021-10-29 安徽大学 一种纳米多孔金及其制备方法和电化学分析传感器

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