CN111537580A - Sensing electrode based on optical fiber bundle and preparation method and application thereof - Google Patents
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Abstract
The invention discloses a sensing electrode based on an optical fiber bundle and a preparation method and application thereof. The micropore bipolar electrode array generates induced polarization potential under the action of an electric field to drive an electrochemiluminescence reaction to be carried out, and a luminescent signal is captured by the optical fiber bundle and then transmitted to the other end, so that the generation and detection of electrochemiluminescence can be separated in space, and the aim of remote sensing is fulfilled; and the micropore bipolar electrode array comprises a large number of independent micropores, so that the area of the electrode on the cross section is increased, and the micropore bipolar electrode array can be used in an electrochemiluminescence sensor to generate a stronger electrochemiluminescence signal.
Description
Technical Field
The invention relates to a sensing electrode based on an optical fiber bundle and a preparation method and application thereof, belonging to the technical field of remote electrochemical luminescence sensing.
Background
Electrochemiluminescence (ECL) is a phenomenon in which a reactant generates an excited state of a micelle through a series of electron transfer reactions on the surface of an electrode, and then transitions to a ground state with light emission. Electrochemiluminescence is widely used in analytical chemistry research, based on the fact that the electrochemiluminescence signal intensity has a direct or indirect linear relationship with the analyte. The electrochemiluminescence analysis has the characteristics of simple and convenient operation of an electrochemical instrument, low background signal, high sensitivity, good spatial resolution of a fluorescence microscope and the like, and provides a more sensitive method for quantitative detection of analytes or determination of surface morphology and properties. The electrochemiluminescence analysis can be used for detecting substances such as heavy metal ions, drug molecules, proteins, nucleic acids, tumor markers and the like. In addition, with the continuous improvement of the electrochemical luminescence detection method and apparatus, the electrochemical luminescence analysis research of particles such as microspheres, nanoparticles, cells and the like has also made remarkable progress, such as the construction of microsphere-based immunosensors, the tracking of nanoparticle collision tracks, the electrocatalytic activity research of gold nanoplates, cell imaging, the analysis of cell membrane proteins and cell secretion content and the like.
Although electrochemiluminescence is widely used in the field of analytical chemistry, its intensity is easily affected by the material and the morphological configuration of the electrode. In addition, the generation of electrochemiluminescence is limited to the solution near the surface of the electrode, and the detector of the luminescence signal usually needs to be placed in front of the electrode at a short distance, which causes problems that the detection is difficult to predict, such as pollution to the sample or the detector due to the close contact, incorrect placement of the detector due to narrow space, and the presence of substances harmful to human body in the detection site. Achieving separation in the generation and detection space of electrochemiluminescence has therefore been one of the interests of scientists in research.
The electrode array with the microporous structure is beneficial to capture and fixation of particle analytes, provides an independent detection unit and is widely used for constructing an electrochemical luminescence sensor; modular planar electrode matrices are commonly used for the preparation and application of micro-porous structured electrode arrays, but are not suitable for further construction of remote electrochemiluminescence sensors. The optical fiber bundle is an excellent carrier of optical signals, consists of a wrapping layer with low refractive index and a quartz glass or plastic core with high refractive index, and is commonly used for remote transmission of different optical signals (from infrared to ultraviolet light waves); however, the optical fiber bundle itself has no conductivity and cannot be directly used for preparing the electrochemical luminescence sensor.
Based on the above, the inventors provide a novel sensing electrode which can be used for remote electrochemiluminescence detection, and construct a remote electrochemiluminescence sensor.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the problems existing in the signal detection process of the existing electrochemical luminescence detection system, the invention provides a sensing electrode based on an optical fiber bundle, a preparation method of the sensing electrode and application of the sensing electrode in remote electrochemical luminescence detection.
The technical scheme is as follows: the invention relates to a sensing electrode based on an optical fiber bundle, which comprises the optical fiber bundle and a protective shell thereof, wherein one end of the optical fiber bundle is wrapped by the shell, the other end of the optical fiber bundle is exposed, the cross section of the exposed end forms a plurality of micropores after the optical fiber bundle is etched, and the cross section and the side surface of the exposed end are coated with transparent conductive films; the optical fiber bundle containing the micropore structure and coated with the conductive film at the exposed end forms a micropore bipolar electrode array.
The number of optical fibers of the bundle and the diameter of the individual optical fibers can be selected according to experimental requirements. Generally speaking, the greater the number of optical fiber bundles, the greater the number of micropores that can be formed, and the stronger the electrochemiluminescence signal; meanwhile, the larger the diameter of a single optical fiber is, the larger the micropores formed after etching is, and the luminous signal can also be increased.
Preferably, the thickness of the conductive film on the cross section of the exposed end is 10-50 nm, and the conductive film is too thin and easy to crack and too thick and has poor light transmittance; further, the conductive thin film may be a gold film, an ITO film, or an FTO film.
The invention relates to a preparation method of a sensing electrode based on an optical fiber bundle, which comprises the following steps:
(1) taking the optical fiber bundle wrapped with the shell, and removing the shell at one end to form an exposed end;
(2) the bare end of the optical fiber bundle is vertically placed at the NH4In the F/HF mixed etching solution, after the optical fiber bundle core is etched, a micropore structure is obtained on the cross section of the exposed end, and the optical fiber bundle core is cleaned and dried by nitrogen for standby;
(3) and rolling the etched optical fiber bundle into a coil, wrapping the rest part except the exposed end with a protective layer, then putting the coil into a coating instrument, and coating a layer of conductive film on the cross section and the side surface of the exposed end to obtain the sensing electrode containing the micropore bipolar electrode array.
In the step (1), one end of the optical fiber bundle can be wiped by acetone to remove the shell of the optical fiber bundle; the bare end can then be cut to the desired length using an optical fiber cutter.
Preferably, NH of step (2)4In a F/HF mixed etching solution, NH4The mass percent concentration of F is 35-50%, the mass percent concentration of HF is 40-50%, and NH4F. The volume ratio of HF to water is 4-6: 0.8-1.2: 1.
The invention relates to an application of a sensing electrode based on an optical fiber bundle in remote electrochemical luminescence detection. Specifically, a remote electrochemiluminescence sensor is constructed, and the electrochemiluminescence sensor comprises a sensing electrode based on a fiber-optic bundle, and a ruthenium terpyridyl complex (Ru (bpy))3 2+) And a co-reactant solution system, a bipolar electrochemical cell and a detector, wherein the sensing electrode based on the optical fiber bundle is provided with a microporous bipolar electrode array at one end and is arranged in the bipolar electrochemical cell, and the other end is positioned outside the bipolar electrochemical cell; the micropore bipolar electrode array generates induced polarization potential under the action of an electric field to drive electrochemical luminescence reaction to proceed, and a luminescence signal is captured by the optical fiber bundle and then transmitted to the other end to be remotely detected by a detector. Preferably, a phosphate buffer solution is used to prepare a solution system containing bipyridyl ruthenium and a coreactant, wherein the coreactant is n-tripropylamine (TPrA) or Nicotinamide Adenine Dinucleotide (NADH).
The invention principle is as follows: the exposed end of one end of the optical fiber bundle is coated with a transparent conductive film and then has conductivity, can generate a luminescent signal in an electrochemical luminescence system, and is captured and transmitted to another optical fiber bundleEnd detection; meanwhile, the GeO is doped due to the different materials of the wrapping layer and the bundle core which form the optical fiber2The quartz glass core is corroded in the etching solution faster than the wrapping layer doped with F, so that the section of the optical fiber bundle is placed in the specific etching solution to obtain a micropore structure, the prepared micropore bipolar electrode array effectively increases the electrode area, and the electron transfer reaction of reactants on the surface of the electrode is promoted, thereby generating a more sensitive signal.
Has the advantages that: compared with the prior art, the invention has the advantages that: (1) the sensing electrode constructed by taking the optical fiber bundle as a main material is combined with a bipolar electrochemical method, and the micropore bipolar electrode array based on the optical fiber bundle can be used for generating electrochemiluminescence without extra wire connection; (2) the sensing electrode based on the optical fiber bundle is etched on the cross section of the optical fiber bundle to form a plurality of micropores, so that the capture and fixation of particle analytes are facilitated, and each micropore simultaneously generates electrochemiluminescence and can be used for high-flux electrochemiluminescence detection; (3) the preparation method of the electrochemiluminescence sensor based on the optical fiber bundle is convenient and rapid, does not need expensive processing instruments, and is suitable for industrial popularization and application.
Drawings
FIG. 1 is a flow chart of the preparation of a sensing electrode based on a fiber optic bundle;
FIG. 2 is a schematic structural diagram of a remote electrochemiluminescence sensor constructed by using the sensing electrode based on the optical fiber bundle of the present invention;
FIG. 3 is a SEM image of a cross-section of an etched end of a fiber optic bundle of example 1;
FIG. 4 shows a sensing electrode based on a fiber optic bundle in Ru (bpy)3 2+Remote electrochemiluminescence intensity in TPrA system;
FIG. 5 shows a sensing electrode based on a fiber optic bundle in Ru (bpy)3 2+Remote in/TPrA System(ii) electrochemiluminescence spectroscopy;
FIG. 6 shows a sensing electrode based on a fiber optic bundle in Ru (bpy)3 2+Remote electrochemiluminescence imaging in the TPrA system;
FIG. 7 shows a sensing electrode based on a fiber optic bundle in Ru (bpy)3 2+Remote electrochemiluminescence intensity in NADH system;
FIG. 8 shows a sensing electrode based on a fiber optic bundle in Ru (bpy)3 2+A linear graph of remote electrochemiluminescence intensity and NADH concentration in the NADH system;
FIG. 9 shows an unetched optical fiber bundle in Ru (bpy)3 2+Remote electrochemiluminescence intensity in the/TPrA system.
Detailed Description
The technical solution of the present invention is further explained with reference to the drawings and the embodiments.
The sensing electrode 7 based on the optical fiber bundle is prepared by taking the optical fiber bundle 1 as a main material, removing the shell 2 at one end, and then carrying out cross section etching, conductive film coating and other treatment as shown in figure 1.
For convenience of explanation, the end face of the exposed end of the optical fiber bundle is defined as a cross section, and the outer surface of the optical fiber bundle of the exposed end is defined as a side face. The sensing electrode 7 based on the optical fiber bundle comprises the optical fiber bundle 1, wherein the optical fiber bundle 1 comprises a bundle core 4 and a wrapping layer 3, and the outside of the optical fiber bundle 1 is wrapped by a protective shell 2; the outer shell 2 at one end of the optical fiber bundle 1 is wrapped and removed, namely one end is exposed, the wrapping layer 3 is directly exposed outside, the cross section of the exposed end is etched to form a large number of micropores 5, and the cross section and the side surface of the exposed end are coated with transparent conductive films 6. The optical fiber bundle which contains a micropore structure and is coated with a conductive film at the exposed end forms a micropore bipolar electrode array, under the action of an electric field, an induced polarization potential is generated to drive an electrochemiluminescence reaction to proceed, and a luminescence signal is captured by the optical fiber bundle and then transmitted to the other end, so that remote electrochemiluminescence detection is realized.
The remote electrochemical luminescence sensor constructed by the sensing electrode 7 based on the optical fiber bundle comprises the sensing electrode 7 based on the optical fiber bundle, two driving electrodes 10, a bipolar electrochemical cell 8, a direct current power supply 9 and a detector 11, wherein the driving electrodes 10 are connected with an external direct current power supply 9, as shown in fig. 2. The process of generating and detecting electrochemiluminescence by using the sensor is as follows:
electrochemical luminescence generation: a sensing electrode 7 based on a fiber optic bundle is placed in a bipolar electrochemical cell 8, a micropore bipolar electrode array at one end of the sensing electrode is positioned in a glass capillary between two driving electrodes 10, and then Ru (bpy) -containing3 2+Adding a solution system of the coreactant into a bipolar electrochemical cell 8, inducing two ends of a sensing electrode 7 to generate a polarization potential under the action of an electric field, wherein Ru (bpy) is contained in the solution3 2+And the co-reactant react with each other to generate Ru (bpy) after the co-reactant is simultaneously oxidized at the induction anode end3 2+Returns to the ground state and emits light;
remote detection of electrochemiluminescence: the electrochemiluminescence generated by the sensing electrode 7 based on the optical fiber bundle is captured by the optical fiber bundle and transmitted to the other end, and signals such as luminous intensity, spectrum and imaging can be remotely detected by using a photomultiplier tube (PMT), an inductively coupled detector (CCD) and the like respectively, so that the detection of remote electrochemiluminescence signals is realized.
Example 1
And (3) intercepting the optical fiber bundle with the length of 2cm to carry out etching and coating experiments, and observing the cross section appearance of the optical fiber bundle after the experiments. The optical fiber bundle specification is: containing 6000 optical fibers, the diameter of each optical fiber is about 4 μm; the etching solution consists of 100 mul of HF solution with the mass percent concentration of 50 percent and 500 mul of NH with the mass percent concentration of 40 percent4Solution F and 100. mu. L H2And O is mixed.
The specific experimental process is as follows:
(1) cutting a section with the length of 2cm on an optical fiber bundle with the length of 50cm by using an optical fiber cutting machine, wiping a black shell of the section with a cotton swab stained with acetone to clean, ultrasonically cleaning for 3min by using deionized water, washing to clean, and drying by using nitrogen for later use;
(2) the 2cm long bundle of optical fibers was vertically set aside by NH4Maintaining the etching solution mixed with F/HF for 5min, taking out to obtain a section of a microporous structure, washing and ultrasonically cleaning the etched optical fiber bundle by using a large amount of deionized water, and blow-drying by using nitrogen for later use;
(3) placing the etched optical fiber bundle on a bracket in a cavity of a coating instrument, coating a gold film for 9min, wiping the cross section of the optical fiber bundle clean by using a polishing cloth, coating the gold film for 3min, washing the optical fiber bundle clean by using deionized water, and drying the optical fiber bundle by using nitrogen;
performing electron microscope scanning operation on the etched surface of the sample with the length of 2cm obtained in the step (3) to obtain an SEM image as shown in FIG. 3; it can be seen that the cross-section of the optical fiber bundle is uniformly distributed with micropores having a diameter of about 2 μm, indicating that the bundle core of the optical fiber bundle is corroded by the etching liquid to form micropores.
Example 2
In this example, a sensing electrode based on a fiber optic bundle was prepared.
The optical fiber bundle specification adopted is as follows: containing 6000 optical fibres, the diameter of the individual optical fibres being approximately 4 μm. The etching solution consists of 100 mul of HF solution with the mass percent concentration of 50 percent and 500 mul of NH with the mass percent concentration of 40 percent4Solution F and 100. mu. L H2And O is mixed.
The preparation process comprises the following steps:
(1) pretreatment of the optical fiber bundle: wiping and cleaning a black shell at one end of an optical fiber bundle with the length of 50cm by using a cotton swab stained with acetone, cutting the black shell into a bare end with the length of 2cm by using an optical fiber cutting machine, finally ultrasonically cleaning the bare end in deionized water for 3min, washing the bare end clean, and drying the bare end for later use by using nitrogen;
(2) etching of the optical fiber bundle: the bare end of the optical fiber bundle is vertically placed at the NH4Keeping the mixed etching solution for 5min, and taking out to obtain a cross section containing a uniform micropore structure, wherein the micropore size is about 2 mu m; washing and ultrasonically cleaning the etched optical fiber bundle by using a large amount of deionized water, and drying the optical fiber bundle by using nitrogen for later use;
(3) gold film coating of optical fiber bundle: rolling the optical fiber bundle with the clean exposed end into a coil with a proper size, wrapping and shaping the coil by using an adhesive tape (the exposed end of the etched optical fiber bundle does not need to be wrapped), putting the coil on a bracket in a cavity of a coating instrument, coating a gold film on the cross section and the side surface of the exposed end for 9min, wiping the cross section by using a polishing cloth, coating the gold film on the cross section and the side surface of the exposed end for 3min, and finally forming the gold films with the thicknesses of about 20nm and 650nm on the cross section and the side surface of the exposed end respectively; and (3) washing the substrate with deionized water, drying the substrate with nitrogen, and preparing a microporous bipolar electrode array at the exposed end, wherein the optical fiber bundle containing the microporous bipolar electrode array is the sensing electrode based on the optical fiber bundle.
Example 3
The optical fiber bundle-based sensing electrode prepared in example 2 was used in Ru (bpy)3 2+The TPrA system is used for carrying out remote electrochemiluminescence test, remote electrochemiluminescence spectrum test and remote electrochemiluminescence imaging test respectively.
(1) Remote electrochemiluminescence testing
As in fig. 2, the fiber optic bundle based sensing electrode 7 is properly installed in the bipolar electrochemical cell 8; 20mL of a solution containing 0.2mM Ru (bpy) was prepared in 10mM phosphate buffer (pH 7.4)3 2+And 20mM TPrA; adding 10mL of test solution into a bipolar electrochemical cell 8, and alternately applying direct current voltages of 0V and 15V to a graphite rod driving electrode 10; at the other end of the sensing electrode 7, a PMT is used for detecting a remote electrochemiluminescence signal; as shown in FIG. 4, no electrochemiluminescence signal was generated when the applied voltage was 0V, and a stable and strong electrochemiluminescence signal was continuously measured when the applied voltage was 15V.
(2) Remote electrochemiluminescence spectroscopy testing
The sensing electrode 7 based on the optical fiber bundle is correctly arranged in the bipolar electrochemical cell 8; 20mL of a solution containing 0.2mM Ru (bpy) was prepared in 10mM phosphate buffer (pH 7.4)3 2+And 20mM TPrA; adding 10mL of test solution into a bipolar electrochemical cell 8, and applying 15V direct-current voltage to a graphite rod driving electrode 10; detecting a remote electrochemiluminescence spectrum by using a CCD spectrometer at the other end of the sensing electrode 7 based on the optical fiber bundle; as shown in FIG. 5, the maximum emission wavelength of the remote electrochemiluminescence spectrum is 623nm, and Ru (bpy)3 2+Substantially uniform in the maximum emission wavelength of the photoluminescence spectrum.
(3) Remote electrochemiluminescence imaging testing
The sensing electrode 7 based on the optical fiber bundle is correctly arranged in the bipolar electrochemical cell 8; 20mL of a solution containing 0.2mM Ru (bpy) was prepared in 10mM phosphate buffer (pH 7.4)3 2+And 20mM TPrA; adding 10mL of test solution into a bipolar electrochemical cell 8, and applying 15V direct-current voltage to a graphite rod driving electrode 10; detecting remote electrochemiluminescence imaging by using a CCD camera at the other end of the sensing electrode 7 based on the optical fiber bundle; as shown in fig. 6, each micro-well in the micro-well bipolar electrode array of the sensing electrode 7 produces a clearly distinguishable remote electrochemiluminescence image.
Example 4
The optical fiber bundle-based sensing electrode prepared in example 2 was used in Ru (bpy)3 2+The NADH system was subjected to remote electrochemiluminescence testing.
The sensing electrode 7 based on the optical fiber bundle is correctly arranged in the bipolar electrochemical cell 8; 20mL of a solution containing 0.1mM Ru (bpy) was prepared in 10mM phosphate buffer (pH 7.4)3 2+And 3mM NADH; adding 10mL of test solution into a bipolar electrochemical cell 8, and alternately applying direct current voltages of 0V and 13V to a graphite rod driving electrode 10; at the other end of the sensing electrode 7, a PMT is used for detecting a remote electrochemiluminescence signal; as a result of the measurement, as shown in FIG. 7, no electrochemiluminescence signal was generated when the applied voltage was 0V, and a stable electrochemiluminescence signal was continuously measured when the applied voltage was 13V.
As shown in FIG. 8, hold Ru (bpy)3 2+Is constant, the remote electrochemiluminescence intensity of the sensing electrode 7 based on the optical fiber bundle is measured to have a linear relation with the concentration of NADH.
Comparative example
A sensing electrode based on an optical fiber bundle was prepared by referring to the method of example 2 except that the etching of the optical fiber bundle in the step (2) was omitted; the finally prepared sensing electrode based on the optical fiber bundle does not have a micropore structure.
Test method according to example 2Method, optical fiber bundle-based sensing electrode (without micro-pore structure) is used for Ru (bpy)3 2+The remote electrochemiluminescence test of the/TPrA system shows the test result as shown in FIG. 9, no electrochemiluminescence signal is generated when the applied voltage is 0V, and a stable electrochemiluminescence signal is continuously measured when the applied voltage is 15V.
Compared with the sensing electrode containing the micropore bipolar electrode array prepared in example 2, the remote electrochemiluminescence signal of the sensing electrode which is not etched in the embodiment is obviously weaker.
Claims (9)
1. A sensing electrode based on an optical fiber bundle is characterized by comprising the optical fiber bundle and a protective shell thereof, wherein one end of the optical fiber bundle is wrapped by the shell, the other end of the optical fiber bundle is exposed, the cross section of the exposed end forms a plurality of micropores after the optical fiber bundle is etched, and the cross section and the side surface of the exposed end are coated with transparent conductive films; and the optical fiber bundle at the exposed end, which contains the micropore structure and is coated with the conductive film, forms a micropore bipolar electrode array.
2. The optical fiber bundle-based sensing electrode according to claim 1, wherein the thickness of the conductive film of the exposed end cross section is 10-50 nm.
3. The fiber optic bundle-based sensing electrode of claim 1, wherein the conductive film is a gold film, an ITO film, or an FTO film.
4. The method for preparing the sensing electrode based on the optical fiber bundle according to claim 1, which is characterized by comprising the following steps:
(1) taking the optical fiber bundle wrapped with the shell, and removing the shell at one end to form an exposed end;
(2) the bare end of the optical fiber bundle is vertically placed at the NH4In the F/HF mixed etching solution, after the optical fiber bundle core is etched, a micropore structure is obtained on the cross section of the exposed end, and the optical fiber bundle core is cleaned and dried by nitrogen for standby;
(3) and rolling the etched optical fiber bundle into a coil, wrapping the rest part except the exposed end with a protective layer, then putting the coil into a coating instrument, and coating a layer of conductive film on the cross section and the side surface of the exposed end to obtain the sensing electrode containing the micropore bipolar electrode array.
5. The method for preparing the sensing electrode based on the optical fiber bundle according to claim 4, wherein in the step (1), the shell at one end of the optical fiber bundle is wiped clean by acetone to obtain an exposed end.
6. The method for preparing the optical fiber bundle-based sensing electrode according to claim 4, wherein in the step (2), the NH is added4In a F/HF mixed etching solution, NH4The mass percent concentration of F is 35-50%, the mass percent concentration of HF is 40-50%, and NH4F. The volume ratio of HF to water is 4-6: 0.8-1.2: 1.
7. Use of the fiber optic bundle-based sensing electrode of claim 1 for remote electrochemiluminescence detection.
8. The use according to claim 7, wherein a remote electrochemiluminescence sensor is constructed comprising a fiber optic bundle-based sensing electrode, a solution system containing a terpyridyl ruthenium complex and a co-reactant, a bipolar electrochemical cell and a detector, wherein the fiber optic bundle-based sensing electrode has a microporous bipolar electrode array at one end mounted in the bipolar electrochemical cell and the other end located outside the bipolar electrochemical cell; the micropore bipolar electrode array generates induced polarization potential under the action of an electric field to drive electrochemical luminescence reaction to be carried out, and a luminescence signal is captured by the optical fiber bundle, then transmitted to the other end and remotely detected by a detector.
9. The use according to claim 8, wherein the solution system containing the ruthenium terpyridyl complex and the coreactant is prepared by using a phosphate buffer solution, wherein the coreactant is n-tripropylamine or nicotinamide adenine dinucleotide.
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Application publication date: 20200814 |