CN-121978179-A - Method for detecting charge transmembrane transport between target organism and material interface

CN121978179ACN 121978179 ACN121978179 ACN 121978179ACN-121978179-A

Abstract

The application relates to a method for detecting charge transmembrane transport between a target organism and a material interface, which comprises the following steps of providing a working electrode, sampling electrolyte containing a target bioactive material on the working surface loaded with the functional material to form micro-droplets with a volume of microliter, inserting a reference electrode and a counter electrode into the micro-droplets to form a micro-droplet electrochemical detection loop together with the working surface, applying an excitation signal to the micro-droplet electrochemical detection loop, collecting a corresponding response signal, and determining charge transmembrane transport characteristics between the target bioactive material and the functional material interface based on the response signal. The application successfully combines a micro-droplet system with an electrochemical detection technology by constructing a micro-droplet electrochemical detection loop, and realizes high-sensitivity, low-noise and quantitative detection of a charge transmembrane transport process between a target bioactive substance and a material interface.

Inventors

LEI YU
GAO JIANXIANG

Assignees

清华大学深圳国际研究生院

Dates

Publication Date: 20260505
Application Date: 20260129

Claims (10)

1. A method for detecting charge transport across a membrane between a target organism and a material interface, comprising the steps of: s1, providing a working electrode, wherein the working electrode is provided with a working surface, and the working surface is provided with a functional material; s2, an electrolyte containing a target bioactive substance is added to a working surface loaded with the functional material, so that a micro-droplet with a micro-upgrading volume is formed on the working surface; S3, inserting the tail ends of a reference electrode and a counter electrode into the micro-droplet, and forming a micro-droplet electrochemical detection loop together with the working surface; s4, applying an excitation signal to the micro-droplet electrochemical detection loop, and collecting a response signal from the working electrode; S5, determining the charge transmembrane transport characteristic between the target bioactive substance and the functionalized material interface based on the response signal.
2. The method of claim 1, wherein the excitation signal comprises at least one of a Cyclic Voltammetry (CV) scan signal, a Differential Pulse Voltammetry (DPV) scan signal, and an Electrochemical Impedance Spectroscopy (EIS) scan signal.
3. The method according to claim 1, wherein in step S1, the functionalized material comprises a two-dimensional material, the two-dimensional material being a material having a thickness on the order of nanometers and a two-dimensional planar structure.
4. The method according to claim 1, wherein in step S2, the target bioactive substance comprises a microorganism, a cell, a virus, an enzyme, a nucleic acid or a protein.
5. The method of claim 4, wherein the microorganism comprises escherichia coli, lactobacillus, or shiva.
6. The method according to claim 2, wherein applying the excitation signal in step S4 comprises the steps of: when adopting cyclic voltammetry, scanning is carried out at a selected scanning rate in a selected potential interval; When differential pulse voltammetry is adopted, scanning is carried out in a selected potential interval by using selected potential step, pulse potential and pulse time parameters; when the electrochemical impedance spectroscopy is adopted, the selected direct current bias voltage and alternating current disturbance potential are overlapped on the basis of the open-circuit potential of the micro-droplet electrochemical detection loop, and scanning is carried out in a selected frequency range.
7. The method of claim 1, wherein the charge transport characteristics across the membrane in step S5 include an interface charge transfer direction, a charge transfer resistance, an electron transfer number, or a target bioactive concentration.
8. A microbial electrochemical sensor for performing the method of any one of claims 1-9 to detect a concentration of a microorganism, the sensor comprising a housing, an electrochemical workstation disposed within the housing for generating and collecting an electrical signal, a detection cell disposed on or in communication with the housing for housing a micro-droplet electrochemical detection circuit, and a processor electrically connected to the electrochemical workstation configured to calculate and output concentration information of the microorganism based on a response signal collected by the electrochemical workstation.
9. A method for evaluating the efficacy of an antibacterial material, which is characterized by comprising the steps of detecting the charge transmembrane transport characteristic between a target antibacterial material and a microorganism interface by using the method according to any one of claims 1 to 9, and evaluating the antibacterial efficacy of the target antibacterial material based on the change of the charge transmembrane transport characteristic, wherein the change of the charge transport process or the difference of charge transfer amounts affects the antibacterial effect.
10. A micro-droplet electrochemical detection device comprises a working electrode, a reference electrode and a counter electrode, wherein the working surface of the working electrode is provided with a functional material, and the tail end of the reference electrode and the tail end of the counter electrode are configured to be inserted into micro-droplets which are formed on the working surface and are micro-updated so as to form a micro-droplet electrochemical detection loop together.

Description

Method for detecting charge transmembrane transport between target organism and material interface Technical Field The invention relates to the field of bioelectrochemical detection, in particular to a method for detecting charge transmembrane transport between a target organism and a substance interface. Background The charge transmembrane transport process between the microorganism and the material interface is a key scientific problem in the fields of environmental microbiology, bioenergy, biomedicine and the like. The accurate analysis process is important to develop a high-efficiency microbial electrochemical sensor, optimize the performance of a microbial fuel cell and evaluate the performance of a novel antibacterial material. Currently, detection techniques in this field rely primarily on macroscopic electrochemical cells (e.g., 50 mL systems). The traditional methods have the following remarkable defects that firstly, the required sample is large in size, precious or low-concentration samples are not detected easily, the cost is high, secondly, a charge transmission path in a macroscopic system is long, diffusion loss in electrolyte is serious, intrinsic signals derived from interface charge transfer are weak, furthermore, irregular diffusion of free ions in solution can generate strong background noise, weak transmembrane charge transfer signals are easy to mask, and finally, the interface process is usually only estimated indirectly through macroscopic output results (such as integral current and voltage), and direct and accurate quantification capability on interface charge transfer kinetic parameters (such as charge transfer rate constant and charge transfer resistance) is lacked, so that microscopic mechanisms of the interface charge transfer kinetic parameters are difficult to reveal. Micro-droplet technology presents unique advantages in biochemical analysis due to its high surface-to-volume ratio, low sample consumption, and limited reaction space. However, combining the micro-droplet system with the electrochemical detection technology and applying the micro-droplet system to the precise quantification of the bio-charge transmembrane transport process still faces technical challenges such as how to stably construct a micro-droplet three-electrode detection loop, how to select a functional material that can specifically interact with a microbial interface to amplify a signal, and how to establish a quantitative relationship between an electrical signal and an interface transport property. Therefore, a new method for realizing high-sensitivity, high-precision and quantitative detection on the charge transmembrane transport process between microbial interfaces is urgently needed in the field. It should be noted that the information disclosed in the above background section is only for understanding the background of the application and thus may include information that does not form the prior art that is already known to those of ordinary skill in the art. Disclosure of Invention The top-layer technical problem to be solved by the invention is how to directly and accurately quantify the charge transmembrane transport kinetic parameters between the target bioactive substances and the material interface, and particularly how to establish a novel method capable of realizing high-sensitivity, low-noise and quantitative detection on the charge transmembrane transport process between the target bioactive substances (such as microorganisms, cells, viruses, enzymes, nucleic acids or proteins) interface so as to overcome the core limitations of weak signals, high background noise, large sample consumption and incapability of directly and accurately quantifying the interface kinetic parameters in the traditional macroscopic electrochemical detection technology. The technical scheme adopted by the application for solving the technical problems is as follows. The application provides a method for detecting charge transmembrane transport between a target organism and a substance interface, which comprises the following steps: s1, providing a working electrode, wherein the working electrode is provided with a working surface, and the working surface is provided with a functional material; S2, applying an electrolyte containing a target bioactive substance to a working surface loaded with a functional material to form a micro-droplet with a volume of micro-upgrading; s3, inserting the tail ends of the reference electrode and the counter electrode into the micro-liquid drops, and forming a micro-liquid drop electrochemical detection loop together with the working surface; s4, applying an excitation signal to the micro-droplet electrochemical detection loop, and collecting a response signal from the working electrode; s5, determining the charge transmembrane transport characteristic between the target bioactive substance and the interface of the functional material based on the response signal. In some embodi