Multi-layer Through-Type Microbattery, Water Quality Detection Method, and Microbial Electrochemical Sensor

Abstract

This application belongs to the technical field of biosensors, and specifically discloses a multi-layer through-type micro-battery, a water quality detection method, and a microbial electrochemical sensor. The multi-layer through-type micro-battery includes: a base, on which a first working electrode circuit interface, a first counter electrode circuit interface, a first reference electrode circuit interface, a water inlet pipe interface, and a first liquid channel extending in the vertical direction are provided; a first working electrode covering the first liquid channel; an intermediate cavity, on which a second working electrode circuit interface, a second counter electrode circuit interface, a second reference electrode circuit interface, and a second liquid channel vertically penetrating the intermediate cavity are provided; a second working electrode covering the second liquid channel; and a top piece provided with a water outlet pipe interface. This application improves the output signal of the detection device, increases the hydraulic retention time to promote the reaction process between the substance to be detected and the electrode, thereby reducing detection interference and improving sensitivity.

Description

Technical Field

[0001] This application belongs to the technical field of biosensors, and particularly relates to a multi-layer through-type micro-battery, a water quality detection method, and a microbial electrochemical sensor. Background Art

[0002] A microbial electrochemical sensor (MES) with electrochemically active bacteria (EAB) as the core can directly convert the substance to be measured in water into a bioelectric signal, and has the advantages of rapid detection, high sensitivity, low detection cost, strong anti-interference ability, etc., and has good application prospects in the fields of biomedicine and environmental monitoring. EAB exists widely in nature, and most of its daily applications are mainly based on the mixture of multi-strain EAB and the formation of a mature mixed bacterial biofilm through long-term laboratory domestication. Dozens of EAB are enriched in the mature mixed bacterial biofilm, with a relatively high electricity generation signal, easy to detect, and strong stress resistance, and can be quickly and automatically repaired after encountering a toxicity shock. It has been widely used in water toxicity detection. However, the various EAB enriched in the mixed bacterial biofilm are extremely prone to dynamic changes due to differences in the domestication environment and test environment (temperature, pH, oxygen, nutrient solution composition, and test water conditions), resulting in problems such as unstable electricity generation performance and a significant change in sensitivity to toxic substances. At the same time, the mature mixed bacterial biofilm also cannot detect low-concentration toxic substances due to its strong stress resistance. Screening a single pure strain of EAB from the mature mixed bacterial biofilm can solve this problem, but pure strain EAB often has weak electricity generation performance, the signal detection process is extremely vulnerable to interference, and pure strain EAB is extremely prone to death, further increasing the detection difficulty. Summary of the Invention

[0003] This application aims to at least solve one of the technical problems in the related art to some extent. For this reason, the purpose of this application is to propose a multi-layer through-type micro-battery, a water quality detection method, and a microbial electrochemical sensor.

[0004] In one aspect of this application, this application proposes a multi-layer through-type micro-battery. According to the embodiments of this application, the multi-layer through-type micro-battery includes:

[0005] A base, with a first groove provided on the upper part of the base, and a first working electrode line interface, a first pair of electrode line interfaces, a first reference electrode line interface and a water inlet interface provided on the side of the base; a first liquid channel extending in the vertical direction is further provided on the base, the first liquid channel penetrates through the first groove in the vertical direction and extends to the water inlet interface, and the first working electrode line interface, the first pair of electrode line interfaces and the first reference electrode line interface are respectively communicated with the first liquid channel;

[0006] A first working electrode, which is arranged in the first groove, covers the first liquid channel, and the orthographic projection of the first working electrode in the vertical direction covers the orthographic projection of the first liquid channel in the vertical direction;

[0007] At least one intermediate cavity, the lower side of the intermediate cavity is connected to the upper side of the base, a second groove is provided on the upper part of the intermediate cavity, and a second working electrode line interface, a second pair of electrode line interfaces and a second reference electrode line interface are provided on the side of the intermediate cavity; a second liquid channel penetrating through the intermediate cavity in the vertical direction is further provided on the intermediate cavity, the second liquid channel penetrates through the second groove in the vertical direction, and the second working electrode line interface, the second pair of electrode line interfaces and the second reference electrode line interface are respectively communicated with the second liquid channel;

[0008] A second working electrode, which is arranged in the second groove, covers the second liquid channel, and the orthographic projection of the second working electrode in the vertical direction covers the orthographic projection of the second liquid channel in the vertical direction;

[0009] A top piece, the lower side of the top piece is connected to the upper side of the intermediate cavity, and a water outlet interface is provided on the top piece, and the water outlet interface is communicated with the second liquid channel.

[0010] For the multi-layer through-type micro-battery according to the embodiment of the present application, by arranging a plurality of working electrodes that can work independently or in series in the same reaction system, the total amount of EAB attached is increased, so that the electrical signal can be effectively improved and external interference can be reduced, solving the problems of low electrical signal and difficult detection of pure bacterial EAB. At the same time, by arranging the first liquid channel and the second liquid channel to form an elongated reaction cavity, the working electrode is clamped between the reaction cavities, and a flow-through detection is adopted to force all the test solution to flow through the working electrode, so as to improve the sensitivity of detecting toxic substances by increasing the contact time between the fixed pure bacterial EAB in the working electrode and the test substance and the fixed liquid path.

[0011] In addition, the multi-layer through-type micro-battery according to the above embodiments of the present application may further have the following additional technical features:

[0012] In some embodiments of the present application, a first working electrode card slot is provided at the bottom of the first groove, and the first working electrode is disposed in the first working electrode card slot; a second working electrode card slot is provided at the bottom of the second groove, and the second working electrode is disposed in the second working electrode card slot.

[0013] In some embodiments of the present application, the multi-layer through-type micro-battery further includes: a first sealing ring and a second sealing ring; a first sealing ring card slot is further provided at the bottom of the first groove, the first sealing ring card slot is disposed outside the first working electrode card slot, and the first sealing ring is disposed in the first sealing ring card slot; a second sealing ring card slot is further provided at the bottom of the second groove, the second sealing ring card slot is disposed outside the second working electrode card slot, and the second sealing ring is disposed in the second sealing ring card slot.

[0014] In some embodiments of the present application, the multi-layer through-type micro-battery further includes: a first working electrode pressing block, the first working electrode pressing block is disposed in the first groove, and the first working electrode pressing block is disposed on the first working electrode and the first sealing ring; a first through hole penetrating the first working electrode pressing block in the vertical direction is provided on the first working electrode pressing block, and the first through hole is directly opposite to the first liquid channel in the vertical direction; a second working electrode pressing block, the second working electrode pressing block is disposed in the second groove, and the second working electrode pressing block is disposed on the second working electrode and the second sealing ring; a second through hole penetrating the second working electrode pressing block in the vertical direction is provided on the second working electrode pressing block, and the second through hole is directly opposite to the second liquid channel in the vertical direction.

[0015] In some embodiments of the present application, the upper part of the first groove is a first internal thread groove, and the first liquid channel penetrates the first internal thread groove in the vertical direction; a first external thread portion is provided at the lower part of the intermediate cavity, the second liquid channel penetrates the first external thread portion in the vertical direction, and the first external thread portion is adapted to the first internal thread groove to connect the base and the intermediate cavity.

[0016] In some embodiments of the present application, the upper part of the second groove is a second internal thread groove, and the second liquid channel penetrates the second internal thread groove in the vertical direction; a second external thread portion is provided at the lower part of the top member, the second liquid channel penetrates the second external thread portion in the vertical direction, and the second external thread portion is adapted to the second internal thread groove to connect the top member and the intermediate cavity.

[0017] In some embodiments of the present application, the first working electrode is a carbon cloth or a carbon felt; the second working electrode is a carbon cloth or a carbon felt.

[0018] In some embodiments of the present application, the diameter of the first liquid channel is (5 mm - 15 mm) / 20 mL of the solution to be measured; the diameter of the second liquid channel is (5 mm - 15 mm) / 20 mL of the solution to be measured.

[0019] In a second aspect of the present application, the present application proposes a method for water quality detection using the multi-layer through-type micro-battery described in the above embodiments. According to the embodiments of the present application, the method includes:

[0020] Introduce the bacterial suspension into the multi-layer through-type micro-battery through the water inlet interface, so that the pure electrochemically active microorganisms in the bacterial suspension are intercepted and attached to the first working electrode and the second working electrode;

[0021] Introduce the basic solution into the multi-layer through-type micro-battery through the water inlet interface, and use a multi-channel potentiostat to collect the initial signal I0 of the multi-layer through-type micro-battery;

[0022] Introduce the solution to be measured into the multi-layer through-type micro-battery through the water inlet interface, and use the multi-channel potentiostat to collect the signal I of the multi-layer through-type micro-battery t ;

[0023] Calculate the inhibition rate of the solution to be measured .

[0024] According to the method for water quality detection using the above multi-layer through-type micro-battery according to the embodiments of the present application, the output signal of the detection device can be greatly improved, and the hydraulic retention time can be increased to promote the reaction process between the substance to be measured and the electrode, thereby reducing detection interference and improving sensitivity.

[0025] In a third aspect of the present application, the present application proposes a microbial electrochemical sensor. According to the embodiments of the present application, the microbial electrochemical sensor includes the multi-layer through-type micro-battery of the above embodiments. Thus, the microbial electrochemical sensor has all the advantages of the multi-layer through-type micro-battery, which will not be elaborated here.

[0026] The additional aspects and advantages of the present application will be partly given in the following description, partly will become obvious from the following description, or be understood through the practice of the present application. Description of the Drawings

[0027] The above and / or additional aspects and advantages of the present application will become obvious and easy to understand from the description of the embodiments in conjunction with the following drawings, where:

[0028] Figure 1 A side schematic view of the multi-layer through-type micro-battery according to an embodiment of the present application;

[0029] Figure 2 Another side schematic view of the multi-layer through-type micro-battery according to an embodiment of the present application;

[0030] Figure 3 A top view of the multi-layer through-type micro-battery according to an embodiment of the present application;

[0031] Figure 4 An exploded schematic view of the multi-layer through-type micro-battery according to an embodiment of the present application;

[0032] Figure 5 A cross-sectional schematic view of the multi-layer through-type micro-battery according to an embodiment of the present application;

[0033] Figure 6 A cross-sectional schematic view of the base according to an embodiment of the present application;

[0034] Figure 7 A cross-sectional schematic view of the intermediate cavity according to an embodiment of the present application;

[0035] Figure 8 A system schematic view of water quality detection using the multi-layer through-type micro-battery according to an embodiment of the present application.

[0036] Reference numerals:

[0037] 1000 - Multi-layer through-type micro-battery, 100 - Base, 101 - First working electrode line interface, 102 - First counter electrode line interface, 103 - First reference electrode line interface, 104 - Water inlet pipe interface, 105 - First groove, 105-1 - First working electrode card slot, 105-2 - First sealing ring card slot, 105-3 - First internal thread groove, 106 - First liquid channel, 110 - First working electrode, 120 - First sealing ring, 130 - First working electrode pressing block, 140 - Third sealing ring, 200 - Intermediate cavity, 201 - Second working electrode line interface, 202 - Second counter electrode line interface, 203 - Second reference electrode line interface, 204 - First external thread portion, 205 - Second groove, 205-1 - Second working electrode card slot, 205-2 - Second sealing ring card slot, 205-3 - Second internal thread groove, 210 - Second working electrode, 220 - Second sealing ring, 230 - Second working electrode pressing block, 300 - Top piece, 301 - Water outlet pipe interface, 302 - Second external thread portion, 2000 - Multi-channel potentiostat, 3000 - Peristaltic pump, 4000 - Solution container. Detailed implementation manners

[0038] Embodiments of the present application will be described in detail below. Examples of the embodiments are shown in the accompanying drawings, where like or similar reference numerals denote like or similar elements or elements having like or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to explain the present application and should not be construed as limiting the present application.

[0039] In the description of the present application, it should be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "axial", "radial", "circumferential", etc. indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings. They are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present application.

[0040] In addition, the terms "first" and "second" are only used for descriptive purposes and should not be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, features defined with "first" and "second" may explicitly or implicitly include at least one of such features. In the description of the present application, "a plurality of" means at least two, such as two, three, etc., unless otherwise specifically and clearly defined.

[0041] In the present application, unless otherwise clearly specified and limited, terms such as "mounted", "connected", "coupled", "fixed", etc. should be understood in a broad sense. For example, it may be a fixed connection, a detachable connection, or integrated; it may be a mechanical connection or an electrical connection; it may be directly connected or indirectly connected through an intermediate medium, and it may be the internal communication of two elements or the interaction relationship between two elements, unless otherwise clearly limited. For those of ordinary skill in the art, the specific meanings of the above terms in the present application can be understood according to specific circumstances.

[0042] In the present application, unless otherwise clearly specified and limited, the first feature being "on" or "under" the second feature may be that the first and second features are in direct contact, or the first and second features are indirectly in contact through an intermediate medium. Moreover, the first feature being "above", "over" and "on top of" the second feature may be that the first feature is directly above or obliquely above the second feature, or merely indicates that the first feature has a higher horizontal height than the second feature. The first feature being "under", "beneath" and "underneath" the second feature may be that the first feature is directly below or obliquely below the second feature, or merely indicates that the first feature has a lower horizontal height than the second feature.

[0043] In one aspect of the present application, a multi-layer through-type micro-battery 1000 is proposed. According to an embodiment of the present application, with reference to the attached Figures 1 - 7 , the above multi-layer through-type micro-battery 1000 includes: a base 100, a first groove 105 is provided on the upper part of the base 100, and a first working electrode line interface 101, a first counter electrode line interface 102, a first reference electrode line interface 103 and a water inlet pipe interface 104 are provided on the side of the base 100; a first liquid channel 106 extending in the vertical direction is further provided on the base 100, the first liquid channel 106 vertically penetrates the first groove 105 and extends to the water inlet pipe interface 104, and the first working electrode line interface 101, the first counter electrode line interface 102 and the first reference electrode line interface 103 are respectively communicated with the first liquid channel 106; a first working electrode 110, the first working electrode 110 is arranged in the first groove 105, the first working electrode 110 covers the first liquid channel 106, and the orthographic projection of the first working electrode 110 in the vertical direction covers the orthographic projection of the first liquid channel 106 in the vertical direction; at least one intermediate cavity 200, the lower side of the intermediate cavity 200 is connected to the upper side of the base 100, a second groove 205 is provided on the upper part of the intermediate cavity 200, and a second working electrode line interface 201, a second counter electrode line interface 202 and a second reference electrode line interface 203 are provided on the side of the intermediate cavity 200; a second liquid channel 206 vertically penetrating the intermediate cavity 200 is further provided on the intermediate cavity 200, the second liquid channel 206 vertically penetrates the second groove 205, and the second working electrode line interface 201, the second counter electrode line interface 202 and the second reference electrode line interface 203 are respectively communicated with the second liquid channel 206; a second working electrode 210, the second working electrode 210 is arranged in the second groove 205, the second working electrode 210 covers the second liquid channel 206, and the orthographic projection of the second working electrode 210 in the vertical direction covers the orthographic projection of the second liquid channel 206 in the vertical direction; a top piece 300, the lower side of the top piece 300 is connected to the upper side of the intermediate cavity 200, and a water outlet pipe interface 301 is provided on the top piece 300, and the water outlet pipe interface 301 is communicated with the second liquid channel 206. Thus, the present application can greatly improve the output signal of the detection device, increase the hydraulic retention time to promote the reaction process between the substance to be detected and the electrode, thereby reducing detection interference and improving sensitivity.

[0044] The beneficial effects that the multi-layer through-type micro-battery proposed by the present application can achieve are described in detail below:

[0045] By arranging multiple working electrodes that can work independently or in series in the same reaction system, the total amount of EAB attached is increased, thereby effectively improving the electrical signal, reducing external interference, and solving the problem of low electrical signal and difficult detection of pure bacterial EAB.

[0046] Meanwhile, in the present application, an elongated reaction cavity is formed by arranging a first liquid channel 106 and a second liquid channel 206, and the working electrode is clamped between the reaction cavities. A flow-through detection is adopted to force all the solution to be measured to flow through the working electrode, so as to improve the sensitivity of detecting toxic substances by increasing the contact time between the pure bacteria EAB immobilized in the working electrode and the substance to be measured and the fixed liquid path. Specifically, the reaction cavity formed by the first liquid channel 106 and the second liquid channel 206 has a small diameter and an extended channel length, presenting an overall shape of an elongated pipe. The multi-layer working electrodes attached with EAB are distributed in the middle of the elongated pipe. When the reaction solution passes through each layer of the working electrode attached with EAB, it penetrates through. Due to the small channel diameter, the solution to be measured can fixedly pass through a limited cross-sectional area.

[0047] According to some specific embodiments of the present application, referring to the attached Figure 6 and 7 , a first working electrode slot 105-1 is provided at the bottom of the first groove 105, and the first working electrode 110 is arranged in the first working electrode slot 105-1; a second working electrode slot 205-1 is provided at the bottom of the second groove 205, and the second working electrode 210 is arranged in the second working electrode slot 205-1.

[0048] According to some other specific embodiments of the present application, referring to the attached Figure 4 , 6 and 7, the above-mentioned multi-layer through-type microbattery further includes: a first sealing ring 120 (such as a fluororubber sealing ring) and a second sealing ring 220 (such as a fluororubber sealing ring); a first sealing ring slot 105-2 is further provided at the bottom of the first groove 105, the first sealing ring slot 105-2 is arranged outside the first working electrode slot 105-1, and the first sealing ring 120 is arranged in the first sealing ring slot 105-2; a second sealing ring slot 205-2 is further provided at the bottom of the second groove 205, the second sealing ring slot 205-2 is arranged outside the second working electrode slot 205-1, and the second sealing ring 220 is arranged in the second sealing ring slot 205-2. The first sealing ring 120 and the second sealing ring 220 can effectively prevent the solution from overflowing.

[0049] According to some other specific embodiments of the present application, referring to the attached Figure 2 and 4, the above multi-layer through-type micro-battery further includes: a first working electrode pressing block 130, the first working electrode pressing block 130 is disposed in the first groove 105, and the first working electrode pressing block 130 is disposed on the first working electrode 110 and the first sealing ring 120, so that the first working electrode pressing block 130 presses the first working electrode 110 and the first sealing ring 120 to prevent them from moving; a first through hole penetrating the first working electrode pressing block 130 in the vertical direction is provided on the first working electrode pressing block 130, and the first through hole is directly opposite to the first liquid channel 106 in the vertical direction; a second working electrode pressing block 230, the second working electrode pressing block 230 is disposed in the second groove 205, and the second working electrode pressing block 230 is disposed on the second working electrode 210 and the second sealing ring 220, so that the second working electrode pressing block 230 presses the second working electrode 210 and the second sealing ring 220 to prevent them from moving; a second through hole penetrating the second working electrode pressing block 230 in the vertical direction is provided on the second working electrode pressing block 230, and the second through hole is directly opposite to the second liquid channel 206 in the vertical direction.

[0050] According to some further specific embodiments of the present application, referring to the attached Figure 6 , the upper part of the first groove 105 is a first internal thread groove 105-3, and the first liquid channel 106 penetrates the first internal thread groove 105-3 in the vertical direction; a first external thread portion 204 is provided at the lower part of the intermediate cavity 200, the second liquid channel 206 penetrates the first external thread portion 204 in the vertical direction, and the first external thread portion 204 is adapted to the first internal thread groove 105-3 to threadedly connect the base 100 and the intermediate cavity 200. The first external thread portion 204 abuts against the first working electrode pressing block 130, and a third sealing ring 140 is provided between the first working electrode pressing block 130 and the first external thread portion 204.

[0051] According to some further specific embodiments of the present application, referring to the attached Figure 7 , the upper part of the second groove 205 is a second internal thread groove 205-3, and the second liquid channel 206 penetrates the second internal thread groove 205-3 in the vertical direction; a second external thread portion 302 is provided at the lower part of the top member 300, the second liquid channel 206 penetrates the second external thread portion 302 in the vertical direction, and the second external thread portion 302 is adapted to the second internal thread groove 205-3 to threadedly connect the top member 300 and the intermediate cavity 200.

[0052] According to some further specific embodiments of the present application, the first working electrode 110 is carbon cloth or carbon felt; the second working electrode 210 is carbon cloth or carbon felt.

[0053] According to some further specific embodiments of the present application, the diameter of the first liquid channel 106 is (5 mm to 15 mm) / 20 mL of the solution to be measured; the diameter of the second liquid channel 206 is (5 mm to 15 mm) / 20 mL of the solution to be measured. Thus, in the present application, by providing the first liquid channel 106 and the second liquid channel 206 to form an elongated reaction cavity, the working electrode is clamped between the reaction cavities, and a flow-through detection is adopted to force all the solution to be measured to flow through the working electrode, so as to improve the sensitivity of detecting toxic substances by increasing the contact time between the pure bacteria EAB immobilized in the working electrode and the substance to be measured and the fixed liquid path.

[0054] As some preferred embodiments, the above multi-layer through-type micro cell includes two intermediate cavities 200. The structures of the two intermediate cavities 200 are the same. Second grooves 205 are respectively provided on the upper parts of the two intermediate cavities 200. Second working electrode line interfaces 201, second counter electrode line interfaces 202 and second reference electrode line interfaces 203 are respectively provided on the sides of the two intermediate cavities 200. Second liquid channels 206 penetrating the intermediate cavities 200 in the vertical direction are further respectively provided on the two intermediate cavities 200. The second liquid channels 206 penetrate the second grooves 205 in the vertical direction, and the second working electrode line interfaces 201, the second counter electrode line interfaces 202 and the second reference electrode line interfaces 203 are respectively communicated with the second liquid channels 206. Then, the above multi-layer through-type micro cell includes two second working electrodes 210. The two second working electrodes 210 are respectively arranged in their corresponding second grooves 205. The second working electrodes 210 cover the second liquid channels 206, and the orthographic projections of the second working electrodes 210 in the vertical direction cover the orthographic projections of the second liquid channels 206 in the vertical direction. Further, the first external thread portion 204 of the first intermediate cavity 200 is adapted to the first internal thread groove 105-3 of the base 100 for connecting the base 100 and the first intermediate cavity 200; the first external thread portion 204 of the second intermediate cavity 200 is adapted to the first internal thread groove 105-3 of the first intermediate cavity 200 for connecting the first intermediate cavity 200 and the second intermediate cavity 200; the second external thread portion 302 of the top member 300 is adapted to the second internal thread groove 205-3 of the second intermediate cavity 200 for connecting the top member 300 and the second intermediate cavity 200.

[0055] In an embodiment of the present application, the first pair of electrode line interfaces 102 are used to connect the counter electrodes of the first electrode group, the first reference electrode line interface 103 is used to connect the reference electrodes of the first electrode group, the first working electrode line interface 101 is used to connect the working electrode related accessories of the first electrode group, the second pair of electrode line interfaces 202 are used to connect the counter electrodes of the second electrode group, the second reference electrode line interface 203 is used to connect the reference electrodes of the second electrode group, and the second working electrode line interface 201 is used to connect the working electrode related accessories of the second electrode group. Then, the above-mentioned reference electrodes, counter electrodes, and working electrode related accessories are further connected to the multi-channel potentiostat 2000, and the overall structure of the test is as shown in the appendix Figure 8 shown.

[0056] In a second aspect of the present application, the present application proposes a method for water quality detection using the multi-layer through-type microbattery of the above embodiment. According to the embodiment of the present application, the above method includes:

[0057] S100: The bacterial suspension enters the multi-layer through-type microbattery 1000 through the water inlet interface 104, so that the pure electrochemically active microorganisms in the bacterial suspension are intercepted and attached to the first working electrode 110 and the second working electrode 210;

[0058] In this step, first, the overall structure after assembling each structural component, electrode group, water inlet pipe, water outlet pipe, and multi-channel potentiostat 2000 (used to collect the detection signals of the microbattery) is as shown in Figure 8 , the appendix Figure 8 In which 4000 represents a solution container for holding various solutions. Then, the bacterial suspension enters the multi-layer through-type microbattery 1000 through the water inlet interface 104, so that the pure electrochemically active microorganisms in the bacterial suspension are intercepted and attached to the first working electrode 110 and the second working electrode 210.

[0059] As a specific embodiment, the water inlet pipe can be placed in the pure EAB bacterial suspension, the peristaltic pump 3000 is started, and the bacterial suspension enters the microbattery and gradually fills the internal cavity layer by layer; the pure EAB in the bacterial suspension is intercepted and attached to the working electrode (i.e., the porous conductive material) by the working electrode, so that the electrical signals generated by the EAB during the test are directly conducted to the working electrode (i.e., the porous conductive material). Then, the microbattery is placed upside down as a whole, the water inlet pipe is suspended, and the peristaltic pump 3000 is continued to be started to drain the bacterial suspension in the middle cavity 200 of the microbattery.

[0060] S200: The basic solution enters the multi-layer through-type microbattery 1000 through the water inlet interface 104, and the multi-channel potentiostat 2000 is used to collect the initial signal I0 of the multi-layer through-type microbattery 1000;

[0061] In this step, place the micro-battery upright again as a whole, put the water inlet pipe into the basic solution (such as DM electrolyte), start the peristaltic pump 3000, the basic solution enters the micro-battery and fills the internal cavity layer by layer, start the multi-channel potentiostat 2000 to collect the detection signal of the micro-battery, denoted as the initial signal I0. Then place the micro-battery upside down again as a whole, the water inlet pipe is suspended, and continue to start the peristaltic pump 3000 to drain the DM electrolyte in the middle cavity 200 of the micro-battery.

[0062] S300: Pass the solution to be measured into the multi-layer through-type micro-battery 1000 through the water inlet interface 104, and use the multi-channel potentiostat 2000 to collect the signal I of the multi-layer through-type micro-battery 1000 t ;

[0063] In this step, place the micro-battery upright again as a whole, put the water inlet pipe into the solution to be measured (such as DM electrolyte containing toxic substances), start the peristaltic pump 3000, the DM electrolyte containing toxic substances enters the micro-battery and fills the internal cavity layer by layer, start the multi-channel potentiostat 2000 to collect the detection signal I of the micro-battery t 。

[0064] S400: Calculate the inhibition rate R of the solution to be measured;

[0065] In this step, calculate the inhibition rate of the solution to be measured 。 The larger the R value, the greater the toxicity intensity of the solution to be measured.

[0066] In this application, multiple electrode groups are set. The electrical signals output by the multiple electrode groups can be collected separately or connected in series for collection. If collected separately, the inhibition rates R of multiple groups can be calculated separately, or the electrical signals collected separately can be added together and then the inhibition rate R can be calculated.

[0067] Specifically, collect three groups of currents separately: the first working electrode 110, the first counter electrode, and the first reference electrode are the first group, and the first group of current is collected; the first second working electrode 210, the first second counter electrode, and the first second reference electrode are the second group, and the second group of current is collected; the second second working electrode 210, the second second counter electrode, and the second second reference electrode are the third group, and the third group of current is collected. Series collection: Connect the first working electrode 110, the first second working electrode 210, and the second second working electrode 210 in series, and then connect the first second counter electrode and the first second reference electrode to collect a group of series currents.

[0068] The method for water quality detection using the above multi-layer through-type micro-battery in this application can greatly improve the output signal of the detection device, increase the hydraulic retention time to promote the reaction process between the substance to be measured and the electrode, thereby reducing detection interference and improving sensitivity.

[0069] In the third aspect of the present application, the present application proposes a microbial electrochemical sensor. According to an embodiment of the present application, the microbial electrochemical sensor includes the multi-layer through-type microbattery of the above embodiment. Thus, the microbial electrochemical sensor has all the advantages of the multi-layer through-type microbattery, which will not be elaborated herein.

[0070] Embodiments of the present application will be described in detail below. It should be noted that the embodiments described below are exemplary and are only used to explain the present application and should not be construed as a limitation of the present application. Additionally, if not explicitly stated, all reagents used in the following embodiments are commercially available or can be synthesized according to the methods described herein or known methods. Reaction conditions not listed are also easily obtainable by those skilled in the art.

[0071] Example 1

[0072] This embodiment provides a multi-layer through-type microbattery. Refer to the attached Figures 1 - 7, the multi-layer through-type micro-battery 1000 includes: a base 100, with a first groove 105 provided on the upper part of the base 100, and a first working electrode circuit interface 101, a first counter electrode circuit interface 102, a first reference electrode circuit interface 103, and a water inlet interface 104 provided on the side of the base 100; a first liquid channel 106 extending in the vertical direction is further provided on the base 100, and the first liquid channel 106 vertically penetrates the first groove 105 and extends to the water inlet interface 104, and the first working electrode circuit interface 101, the first counter electrode circuit interface 102, and the first reference electrode circuit interface 103 are respectively communicated with the first liquid channel 106; a first working electrode 110 (a conductive carbon felt with a thickness of 2 mm), the first working electrode 110 is arranged in the first groove 105, the first working electrode 110 covers the first liquid channel 106, and the orthographic projection of the first working electrode 110 in the vertical direction covers the orthographic projection of the first liquid channel 106 in the vertical direction; two intermediate cavities 200, the structures of the two intermediate cavities 200 are the same, second grooves 205 are respectively provided on the upper parts of the two intermediate cavities 200, and second working electrode circuit interfaces 201, second counter electrode circuit interfaces 202, and second reference electrode circuit interfaces 203 are respectively provided on the sides of the two intermediate cavities 200; second liquid channels 206 vertically penetrating the intermediate cavities 200 are further respectively provided on the two intermediate cavities 200, the second liquid channels 206 vertically penetrate the second grooves 205, and the second working electrode circuit interfaces 201, the second counter electrode circuit interfaces 202, and the second reference electrode circuit interfaces 203 are respectively communicated with the second liquid channels 206. The above multi-layer through-type micro-battery includes two second working electrodes 210 (conductive carbon felts with a thickness of 2 mm), the two second working electrodes 210 are respectively arranged in their corresponding second grooves 205, the second working electrodes 210 cover the second liquid channels 206, and the orthographic projection of the second working electrodes 210 in the vertical direction covers the orthographic projection of the second liquid channels 206 in the vertical direction. The first external thread portion 204 of the first intermediate cavity 200 is adapted to the first internal thread groove 105-3 of the base 100 for connecting the base 100 and the first intermediate cavity 200; the first external thread portion 204 of the second intermediate cavity 200 is adapted to the first internal thread groove 105-3 of the first intermediate cavity 200 for connecting the first intermediate cavity 200 and the second intermediate cavity 200; the second external thread portion 302 of the top piece 300 is adapted to the second internal thread groove 205-3 of the second intermediate cavity 200 for connecting the top piece 300 and the second intermediate cavity 200. The diameter of the first liquid channel 106 is 7 mm; the diameter of the second liquid channel 206 is 7 mm.

[0073] A first working electrode slot 105-1 is provided at the bottom of the first groove 105, and the first working electrode 110 is disposed in the first working electrode slot 105-1; a second working electrode slot 205-1 is provided at the bottom of the second groove 205, and the second working electrode 210 is disposed in the second working electrode slot 205-1. The above-mentioned multi-layer through-type micro-battery further includes: a first sealing ring 120 (fluororubber sealing ring) and a second sealing ring 220 (fluororubber sealing ring); a first sealing ring slot 105-2 is further provided at the bottom of the first groove 105, the first sealing ring slot 105-2 is disposed outside the first working electrode slot 105-1, and the first sealing ring 120 is disposed in the first sealing ring slot 105-2; a second sealing ring slot 205-2 is further provided at the bottom of the second groove 205, the second sealing ring slot 205-2 is disposed outside the second working electrode slot 205-1, and the second sealing ring 220 is disposed in the second sealing ring slot 205-2.

[0074] The above-mentioned multi-layer through-type micro-battery further includes: a first working electrode pressing block 130, the first working electrode pressing block 130 is disposed in the first groove 105, and the first working electrode pressing block 130 is disposed on the first working electrode 110 and the first sealing ring 120, so that the first working electrode pressing block 130 presses the first working electrode 110 and the first sealing ring 120 to prevent them from moving; a first through hole penetrating the first working electrode pressing block 130 in the vertical direction is provided on the first working electrode pressing block 130, and the first through hole is directly opposite to the first liquid channel 106 in the vertical direction; a second working electrode pressing block 230, the second working electrode pressing block 230 is disposed in the second groove 205, and the second working electrode pressing block 230 is disposed on the second working electrode 210 and the second sealing ring 220, so that the second working electrode pressing block 230 presses the second working electrode 210 and the second sealing ring 220 to prevent them from moving; a second through hole penetrating the second working electrode pressing block 230 in the vertical direction is provided on the second working electrode pressing block 230, and the second through hole is directly opposite to the second liquid channel 206 in the vertical direction.

[0075] The method for water quality detection using the above-mentioned multi-layer through-type micro-battery includes the following steps:

[0076] 1) Assemble each structural component, electrode group, water inlet pipe, water outlet pipe, and multi-channel potentiostat 2000 (used to collect the detection signals of the micro-battery). The assembled overall structure is as Figure 8 .

[0077] 2) Place the water inlet pipe in the pure EAB (Shewanella PV-4) bacterial suspension, start the peristaltic pump 3000, and the bacterial suspension enters the micro-battery and gradually fills the internal liquid channels; the pure EAB in the bacterial suspension is intercepted and attached to the porous conductive material of the working electrode, so that the electrical signals generated by the EAB during the test are directly conducted to the conductive material.

[0078] 3) Invert the entire micro-battery, leave the water inlet pipe suspended, and continue to start the peristaltic pump 3000 to drain the bacterial suspension in the middle cavity 200 of the micro-battery.

[0079] 4) Place the entire micro-battery upright again, place the water inlet pipe in the DM electrolyte, start the peristaltic pump 3000, the DM electrolyte enters the micro-battery and gradually fills the internal liquid channels, start the multi-channel potentiostat 2000 to collect the detection signals of the micro-battery, and separately collect three groups of currents, denoted as the initial signals I 01 、I 02 、I 03 。Separate three groups of currents are collected: the first working electrode 110, the first counter electrode, and the first reference electrode are the first group, and the first group of current I 01 is obtained; the first second working electrode 210, the first second counter electrode, and the first second reference electrode are the second group, and the second group of current I 02 is obtained; the second second working electrode 210, the second second counter electrode, and the second second reference electrode are the third group, and the third group of current I 03 is obtained.

[0080] 5) Invert the entire micro-battery again, leave the water inlet pipe suspended, and continue to start the peristaltic pump 3000 to drain the DM electrolyte in the middle liquid channel of the micro-battery.

[0081] 6) Place the entire micro-battery upright again, place the water inlet pipe in the DM electrolyte containing toxic substances, start the peristaltic pump 3000, the DM electrolyte containing toxic substances enters the micro-battery and gradually fills the internal liquid channels, start the multi-channel potentiostat 2000 to collect the detection signals of the micro-battery, and separately collect three groups of currents, denoted as the toxicity signals I t1 、I t2 、I t3 。

[0082] 7) Calculate the toxicity substance inhibition rate 、 、 , the larger the R value, the greater the toxicity intensity of the toxic substance. The results are shown in Table 1.

[0083] Composition of DM electrolyte: Each liter of the solution contains 2.5 g of NaHCO3, 1.0 g of NH4Cl, 0.08 g of CaCl2·H2O, 0.2 g of MgCl2·6H2O, 10 g of NaCl, 7.2 g of HEPES, and 2 mmol of sodium lactate.

[0084] Toxic DM electrolyte: Each liter of the solution contains 2.5 g of NaHCO3, 1.0 g of NH4Cl, 0.08 g of CaCl2·H2O, 0.2 g of MgCl2·6H2O, 10 g of NaCl, 7.2 g of HEPES, 2 mmol of sodium lactate, and 0.2 mg of Hg(NO3)2.

[0085] Table 1

[0086]

[0087] Example 2

[0088] In this example, the electrical signal acquisition method was changed to series acquisition: The first working electrode, the first second working electrode, and the second second working electrode were connected in series, and then connected to the first second pair of electrodes and the first second reference electrode to collect a set of series currents. All other steps were the same as in Example 1.

[0089] 4) The operation was the same as in Example 1, and a set of currents was collected in series, denoted as the initial signal I 04 , and the results are shown in Table 2.

[0090] 6) The operation was the same as in Example 1, and a set of currents was collected in series, denoted as the toxicity signal I t4 , and the results are shown in Table 2.

[0091] Table 2

[0092]

[0093] Result analysis:

[0094] In Example 1, the sum of the three sets of currents collected separately was 28.73 µA + 25.68 µA + 20.86 µA = 75.27 µA, which was slightly higher than the current of 62.91 µA collected in series in Example 2. This shows that there will be certain differences between the two electrical signal acquisition methods, which may be caused by different internal resistances of the battery due to the difference in electrode distance in the two acquisition methods. However, the initial signals of both are within the test range and can be used for testing; the toxicity inhibition rates are both around 30%, and both have obvious toxicity effects, indicating that both testing methods are feasible.

[0095] In the description of this specification, the description with reference to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0096] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present application. Those of ordinary skill in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present application.

Claims

1. A multi-layer through-type micro-battery for a microbial electrochemical sensor, characterized in that, Comprising: A base, on the upper part of which there is a first groove, and on the side of which there are a first working electrode circuit interface, a first pair of electrode circuit interfaces, a first reference electrode circuit interface and a water inlet pipe interface; on the base there is also a first liquid channel extending in the vertical direction, the first liquid channel penetrating through the first groove in the vertical direction and extending to the water inlet pipe interface, and the first working electrode circuit interface, the first pair of electrode circuit interfaces and the first reference electrode circuit interfaces are respectively communicated with the first liquid channel; A first working electrode, which is arranged in the first groove, covers the first liquid channel, and the orthographic projection of the first working electrode in the vertical direction covers the orthographic projection of the first liquid channel in the vertical direction; Two intermediate cavities, the lower side of the first intermediate cavity is connected to the upper side of the base, the lower side of the second intermediate cavity is connected to the upper side of the first intermediate cavity, on the upper part of each intermediate cavity there is a second groove, and on the side of each intermediate cavity there are a second working electrode circuit interface, a second pair of electrode circuit interfaces and a second reference electrode circuit interface; on each intermediate cavity there is also a second liquid channel penetrating through the intermediate cavity in the vertical direction, the second liquid channel penetrating through the second groove in the vertical direction, and the second working electrode circuit interface, the second pair of electrode circuit interfaces and the second reference electrode circuit interfaces are respectively communicated with the second liquid channel; Two second working electrodes, each of which is arranged in the second groove of each intermediate cavity, covers the second liquid channel, and the orthographic projection of each second working electrode in the vertical direction covers the orthographic projection of the second liquid channel in the vertical direction; A top piece, the lower side of which is connected to the upper side of the second intermediate cavity, and on which there is a water outlet pipe interface, and the water outlet pipe interface is communicated with the second liquid channel of the second intermediate cavity; The first working electrode is carbon cloth or carbon felt; the second working electrode is carbon cloth or carbon felt.

2. The multi-layer through-type micro-battery according to claim 1, characterized in that, At the bottom of the first groove there is a first working electrode card slot, and the first working electrode is arranged in the first working electrode card slot; At the bottom of the second groove there is a second working electrode card slot, and the second working electrode is arranged in the second working electrode card slot.

3. The multi-layer through-type micro battery according to claim 2, wherein, Further comprising: A first sealing ring and a second sealing ring; At the bottom of the first groove there is also a first sealing ring card slot, which is arranged outside the first working electrode card slot, and the first sealing ring is arranged in the first sealing ring card slot; At the bottom of the second groove there is also a second sealing ring card slot, which is arranged outside the second working electrode card slot, and the second sealing ring is arranged in the second sealing ring card slot.

4. The multi-layer through-type micro-battery according to claim 3, characterized in that Further comprising: The first working electrode pressing block is arranged in the first groove and is disposed on the first working electrode and the first sealing ring; a first through hole penetrating the first working electrode pressing block in the vertical direction is provided on the first working electrode pressing block, and the first through hole is directly opposite to the first liquid channel in the vertical direction; The second working electrode pressing block is arranged in the second groove and is disposed on the second working electrode and the second sealing ring; a second through hole penetrating the second working electrode pressing block in the vertical direction is provided on the second working electrode pressing block, and the second through hole is directly opposite to the second liquid channel in the vertical direction.

5. The multi-layer through-type micro-battery according to claim 1, characterized in that, The upper part of the first groove is a first internal thread groove, and the first liquid channel penetrates the first internal thread groove in the vertical direction; A first external thread part is provided at the lower part of the first intermediate cavity, and the second liquid channel of the first intermediate cavity penetrates the first external thread part in the vertical direction. The first external thread part is adapted to the first internal thread groove to connect the base and the first intermediate cavity.

6. The multi-layer through-type micro-battery according to claim 1, wherein The upper part of the second groove of the second intermediate cavity is a second internal thread groove, and the second liquid channel of the second intermediate cavity penetrates the second internal thread groove in the vertical direction; A second external thread part is provided at the lower part of the top piece, and the second liquid channel penetrates the second external thread part in the vertical direction. The second external thread part is adapted to the second internal thread groove of the second intermediate cavity to connect the top piece and the second intermediate cavity.

7. A method for water quality detection using the multi-layer through-type microbattery described in any one of claims 1 to 6, characterized in that, Comprising: The bacterial suspension enters the multi-layer through-type micro-battery through the water inlet interface so that the pure electrochemically active microorganisms in the bacterial suspension are intercepted and attached to the first working electrode and the second working electrode; The basic solution enters the multi-layer through-type micro-battery through the water inlet interface, and the initial signal I0 of the multi-layer through-type micro-battery is collected by a multi-channel potentiostat; The solution to be measured enters the multi-layer through-type micro-battery through the water inlet interface, and the multi-channel potentiostat is used to collect the signal I of the multi-layer through-type micro-battery t ; Calculate the inhibition rate of the solution to be measured .

8. A microbial electrochemical sensor, characterized in that, Comprising the multi-layer through-type micro-battery according to any one of claims 1 to 6.

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