CN112932477A - Blood glucose sensor implanted in interocular scleral interstitial fluid of live fish eyeball and preparation method thereof - Google Patents
Blood glucose sensor implanted in interocular scleral interstitial fluid of live fish eyeball and preparation method thereof Download PDFInfo
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Abstract
The invention provides a blood glucose sensor implanted in the liquid between eyeball and sclera tissues of a live fish and a preparation method thereof, aiming at solving the problem that the blood glucose concentration in the live fish body is difficult to stably measure in real time by implanting the blood glucose sensor into the fish body in the prior art. The blood glucose sensor includes: the sensor comprises a sensor base material and a wireless transmission device electrically connected with the sensor base material, wherein the sensor base material comprises a working electrode and a reference electrode, one end part of the working electrode is provided with an induction cavity, an insulating layer is coated between the other end part of the working electrode and the induction cavity, and the outer surface of the insulating layer is coated with the reference electrode; the reference electrode is composed of a copper wire on the outer surface of the insulating layer and a silver/silver chloride slurry layer covering the copper wire; the external surface of the sensing cavity of the working electrode is covered with a perfluorosulfonic acid polymer membrane, and the external surface of the perfluorosulfonic acid polymer membrane is covered with a glucose oxidase layer. The blood glucose sensor has sensitive blood glucose detection accuracy and portability, and has good stability in monitoring blood glucose in the interstitial fluid of eyeball sclera tissues of the implanted live fish.
Description
The application is a divisional application with the application date of 2015, 10 and 19, the application number of 201510680579.5 and the name of 'blood glucose sensor planted in interscleral tissue of eyeball and sclera of live fish and a preparation method', and is named as Chinese agriculture university.
Technical Field
The invention relates to a bioelectrochemical detection technology, in particular to a blood glucose sensor planted in the interstitial fluid of eyeball and sclera tissues of a live fish and a preparation method thereof.
Background
The fresh and alive fish is favored by consumers because of delicious taste and high nutritive value and is rich in high-quality protein, vitamins and trace elements. The blood sugar level of the live fish is an important index reflecting the stress level of the live fish in the transportation process. The real-time wireless monitoring and acquisition of the blood sugar level of the live fish is a reference basis for continuously improving the live-keeping transportation technology and equipment of the fresh and live fish; meanwhile, the live fish is implanted in a fish body in a minimally invasive manner by means of a biosensing technology and a wireless communication technology, and the technology for monitoring different stress blood sugar levels of the live fish in each stage in the transportation process in real time is also an important information acquisition technology for further optimizing the keep-alive transportation process. However, when the blood glucose sensor is directly implanted into a living fish body, blood protein coagulation occurs at the sensing part of the sensor, thereby reducing or inhibiting the output of the sensing current of the sensor. Therefore, it is difficult to stably measure the blood glucose concentration in the live fish body in real time by directly implanting the sensor into the fish body.
Disclosure of Invention
The invention provides a blood glucose sensor implanted in the liquid between eyeball and sclera tissues of a live fish and a preparation method thereof, which are used for solving the problem that the blood glucose concentration in the live fish body is difficult to stably measure in real time by implanting the blood glucose sensor into the fish body in the prior art.
In order to solve the above problems:
in one aspect, the present invention provides a blood glucose sensor implanted in the interstitial fluid of the sclera tissue of a live fish eyeball, comprising: the sensor comprises a sensor substrate and a wireless transmission device electrically connected with the sensor substrate and used for transmitting current signals, wherein the sensor substrate comprises a working electrode and a reference electrode, one end of the working electrode is provided with an induction cavity, an insulating layer covers the surface of the working electrode between the other end of the working electrode and the induction cavity, and the reference electrode covers the outer surface of the insulating layer;
the reference electrode is composed of copper wires spirally wound on the outer surface of the insulating layer at intervals and a silver/silver chloride slurry layer paved to cover the copper wires; the outer surface of the sensing cavity of the working electrode is covered with a perfluorosulfonic acid polymer membrane, and the outer surface of the perfluorosulfonic acid polymer membrane is covered with a glucose oxidase layer.
Further, an insulating heat-shrinkable tube is arranged on the outer surface of the reference electrode.
Furthermore, the working electrode is a platinum iridium working electrode, and the insulating layer on the outer surface of the working electrode is a polytetrafluoroethylene layer.
Furthermore, an epoxy resin layer is coated at the outer end of the induction cavity of the working electrode.
Further, the wireless transmission device comprises an electrochemical signal amplifier, a wireless transmission module and an on-board antenna, wherein the electrochemical signal amplifier is respectively and electrically connected with the working electrode and the reference electrode through electrode leads; the wireless transmission module is electrically connected with the electrochemical signal amplifier; the onboard antenna is electrically connected with the wireless transmission module.
Further, the diameter of the epoxy resin layer is 0.175mm-0.2mm, and the length is 0.3mm-0.4 mm.
Further, the length of the perfluorinated sulfonic acid polymer membrane is 1mm-1.1 mm.
Further, the diameter of the epoxy resin layer is 0.175mm, and the length is 0.3 mm.
Further, the length of the perfluorosulfonic acid polymer membrane is 1 mm.
In another aspect, the present invention provides a method for manufacturing a blood glucose sensor, comprising the steps of:
adopting platinum-iridium alloy materials to manufacture a working electrode, setting one end part of the manufactured working electrode as an induction cavity, and covering a polytetrafluoroethylene layer on the surface of the working electrode between the other end part of the working electrode and the induction cavity;
winding copper wires on the polytetrafluoroethylene layer area of the working electrode in an interval spiral mode, and covering the copper wires with a silver/silver chloride slurry layer to form a reference electrode;
coating an epoxy resin layer on the outer end of the induction cavity, coating 5% -6% of perfluorosulfonic acid polymer solution on the surface of the residual area of the induction cavity between the outer end of the induction cavity and the polytetrafluoroethylene layer, and air-drying to form a perfluorosulfonic acid polymer film;
soaking the working electrode coated with the perfluorosulfonic acid polymer membrane in 0.25-3ml of phosphate buffer solution containing 2.5-3mg of glucose oxidase and dissolved with 5-7mg of bovine serum albumin, wherein the concentration of the phosphate buffer solution is 0.1mol/L, drying in the air for 10-15 minutes, and repeating the process at least twice;
immersing the sensing cavity of the working electrode into 25% -30% glutaraldehyde solution for 5-6 h;
an electrochemical signal amplifier is respectively and electrically connected with the working electrode and the reference electrode through electrode leads;
electrically connecting a wireless transmission module with the electrochemical signal amplifier;
and electrically connecting an onboard antenna with the wireless transmission module.
From the above, it can be seen that: according to the invention, the perfluorosulfonic acid polymer film and the glucose oxidase layer have excellent conductivity, so that the detection efficiency of the blood glucose sensor can be improved. By means of the function of the glutaraldehyde crosslinking agent, the enzyme molecules are effectively fixed and have long service life. In addition, the perfluorinated sulfonic acid polymer membrane is thin and has selective permeability, so that the material transmission resistance and the resistance of the electrode are reduced, and the utilization rate of the working electrode can be greatly improved. Meanwhile, the perfluorinated sulfonic acid polymer membrane can allow hydrogen ions to pass through after absorbing water, has high selective permeability on cations but has a repulsive effect on anions, and has a certain protection effect on electrodes. In the aspect of a reference electrode, the blood glucose sensor can well sense a bioelectricity signal by utilizing the wound copper wire, so that subsequent current signal transmission is facilitated.
Drawings
FIG. 1 is a schematic structural view of a blood glucose sensor according to embodiment 1 of the present invention;
FIG. 2 is a schematic view of the structure of the end of the working electrode of FIG. 1;
FIG. 3 is a schematic structural diagram of a blood glucose sensor according to embodiment 2 of the present invention
FIG. 4 is a graph showing the correlation between the blood glucose concentration in the interstitial fluid of the sclera tissue of a live fish eyeball and the output current of a sensor;
FIG. 5 is a graph showing the correlation between blood concentration of fish and blood glucose concentration of interstitial fluid (ESIF) in sclera tissue of eyeball.
In the figure: 1. a working electrode; 2. a polytetrafluoroethylene layer; 3. a silver/silver chloride paste layer; 4. a copper wire; 5. an electrode lead; 6. an electrochemical signal amplifier; 7. a wireless transmission module; 8. an on-board antenna; 9. a perfluorosulfonic acid polymer membrane; 10. a glucose oxidase layer; 11. an epoxy resin layer; 12. insulating the heat shrinkable tube.
Detailed Description
The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Fig. 1 and 2 show a blood glucose sensor implanted in interstitial fluid between eyeball and sclera tissues of a live fish according to embodiment 1 of the present invention, including: the sensor substrate, and with the sensor substrate electricity is connected and is used for transmitting current signal's wireless transmission device, wherein, the sensor substrate includes working electrode 1 and reference electrode one end of working electrode sets up to the response chamber, follows another tip of working electrode covers the insulating layer of establishing to the working electrode surface between the response chamber and is polytetrafluoroethylene layer 2, has wrapped up at the insulating layer surface and has had the reference electrode. The reference electrode is composed of copper wires 4 spirally wound on the outer surface of the insulating layer at intervals and a silver/silver chloride slurry layer 3 paved to cover the copper wires; the outer surface of the sensing cavity of the working electrode is covered with a perfluorosulfonic acid polymer membrane 9, and the outer surface of the perfluorosulfonic acid polymer membrane is covered with a glucose oxidase layer 10.
An epoxy resin layer 11 is coated at the outer end of the induction cavity of the working electrode, and a perfluorosulfonic acid polymer membrane 9 is coated on the surface of the residual area of the induction cavity between the outer end of the induction cavity and the polytetrafluoroethylene layer. The epoxy resin layer has anti-pollution capacity and has a certain protection effect on the sensor. Wherein, the diameter of the epoxy resin layer can be 0.175mm-0.2mm, and the length is 0.3mm-0.4 mm. The length of the perfluorinated sulfonic acid polymer membrane is 1mm-1.1 mm.
The wireless transmission device comprises an electrochemical signal amplifier 6, a wireless transmission module 7 and an onboard antenna 8, wherein the electrochemical signal amplifier is respectively and electrically connected with a working electrode and a reference electrode through an electrode lead 5; the wireless transmission module is electrically connected with the electrochemical signal amplifier; the onboard antenna is electrically connected with the wireless transmission module.
The blood sugar sensor is implanted into the liquid between eyeball and sclera tissues of the live fish through a minimally invasive technology in the using process, the glucose oxidase layer on the sensing cavity of the working electrode and the blood sugar in the liquid between the eyeball and the sclera tissues generate a bioelectrochemical reaction, and weak current generated by the reaction is amplified by the electrochemical signal amplifier and then effective current information is transmitted in a wireless transmission mode.
The relation between the blood sugar of the interscleral fluid of the fisheye sclera and the output current of the sensor and the obtained relation are determined according to a plurality of experimental results. FIG. 4 is a graph of the relationship between the results obtained in an experimental procedure. From FIG. 4, a linear regression relationship is summarized as: y is 5.414+0.05962X, where the correlation coefficient is R is 0.994, X is the glucose concentration, and Y is the sensor output current. Therefore, the glucose concentration in the interstitial fluid of the sclera tissue of the fish eyeball can be estimated according to the output current of the sensor by the relational expression. From a biological point of view, since the glucose concentration in the interstitial fluid of the fish eyeball scleral tissue and the glucose concentration in the fish body are closely related, as shown in fig. 5, the relationship can also be expressed by linear regression as Y ═ 0.2996+0.9439X, where the correlation coefficient is R ═ 0.961, the sample point N of the test is 111, X is the blood glucose concentration in the fish body, and Y is the blood glucose concentration in the interstitial fluid of the eyeball scleral tissue. Therefore, the blood glucose concentration of the fish body can be estimated by the above relation.
In the embodiment, the detection efficiency of the blood glucose sensor is improved by adopting the excellent conductivity between the perfluorosulfonic acid polymer film and the glucose oxidase layer. By means of the function of the glutaraldehyde crosslinking agent, the enzyme molecules are effectively fixed and have long service life. In addition, the perfluorinated sulfonic acid polymer membrane is thin and has selective permeability, so that the material transmission resistance and the resistance of the electrode are reduced, and the utilization rate of the working electrode can be greatly improved. Meanwhile, the perfluorinated sulfonic acid polymer membrane can allow hydrogen ions to pass through after absorbing water, has high selective permeability on cations but has a repulsive effect on anions, and has a certain protection effect on electrodes. In the aspect of a reference electrode, the blood glucose sensor can well sense a bioelectricity signal by utilizing the wound copper wire, so that subsequent current signal transmission is facilitated.
Fig. 3 shows a blood glucose sensor implanted in the interstitial fluid of the sclera tissue of a live fish eyeball according to embodiment 2 of the present invention. The present embodiment is different from embodiment 1 in that: an insulating heat shrinkable tube 12 is provided on the outer surface of the reference electrode. The insulating heat shrinkable tube can prevent the erosion damage of interstitial fluid to the sensor components.
The method for manufacturing a blood glucose sensor according to embodiment 4, comprising the steps of:
adopting platinum-iridium alloy materials to manufacture a working electrode, setting one end part of the manufactured working electrode as an induction cavity, and covering a polytetrafluoroethylene layer on the surface of the working electrode between the other end part of the working electrode and the induction cavity;
winding copper wires on the polytetrafluoroethylene layer area of the working electrode in an interval spiral mode, and covering the copper wires with a silver/silver chloride slurry layer to form a reference electrode;
coating an epoxy resin layer on the outer end of the induction cavity, coating 5-6% of perfluorosulfonic acid polymer solution on the surface of the rest area of the induction cavity between the outer end of the induction cavity and the polytetrafluoroethylene layer, and air-drying to form a perfluorosulfonic acid polymer film;
soaking the working electrode coated with the perfluorosulfonic acid polymer membrane in 0.25-3ml of phosphate buffer solution containing 2.5-3mg of glucose oxidase and dissolved with 5-7mg of bovine serum albumin, wherein the concentration of the phosphate buffer solution is 0.1mol/L, drying in the air for 10-15 minutes, and repeating the process at least twice;
immersing the working electrode induction cavity into 25-30% glutaraldehyde solution for 5-6 h;
an electrochemical signal amplifier is respectively and electrically connected with the working electrode and the reference electrode through electrode leads;
electrically connecting a wireless transmission module with the electrochemical signal amplifier;
and electrically connecting an onboard antenna with the wireless transmission module.
The steps are used for preparing the blood sugar sensor implanted in the interscleral interstitial fluid of the eyeball and the sclera of the live fish.
Furthermore, those skilled in the art will appreciate that while some embodiments described herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the following claims, any of the claimed embodiments may be used in any combination.
It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim. The word "comprising" does not exclude the presence of elements or steps not listed in a claim. The word "a" or "an" preceding an element does not exclude the presence of a plurality of such elements. The invention may be implemented by means of hardware comprising several distinct elements, and by means of a suitably programmed computer. In the unit claims enumerating several means, several of these means may be embodied by one and the same item of hardware. The usage of the words first, second and third, etcetera do not indicate any ordering. These words may be interpreted as names.
Those of ordinary skill in the art will understand that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit of the corresponding technical solutions and scope of the present invention as defined in the appended claims.
Claims (7)
1. A blood glucose sensor implanted in interstitial fluid between eyeball and sclera tissues of a live fish, comprising: the sensor comprises a sensor substrate and a wireless transmission device electrically connected with the sensor substrate and used for transmitting current signals, wherein the sensor substrate comprises a working electrode and a reference electrode, one end of the working electrode is provided with an induction cavity, an insulating layer covers the surface of the working electrode between the other end of the working electrode and the induction cavity, and the reference electrode covers the outer surface of the insulating layer;
the reference electrode is composed of copper wires spirally wound on the outer surface of the insulating layer at intervals and a silver/silver chloride slurry layer paved to cover the copper wires; the external surface of the sensing cavity of the working electrode is covered with a perfluorosulfonic acid polymer membrane, and the external surface of the perfluorosulfonic acid polymer membrane is covered with a glucose oxidase layer;
an epoxy resin layer is covered at the outer end of the induction cavity of the working electrode; the diameter of the epoxy resin layer is 0.175mm-0.2mm, and the length is 0.3mm-0.4 mm;
the length of the perfluorinated sulfonic acid polymer membrane is 1mm-1.1 mm.
2. The blood glucose sensor of claim 1, wherein an insulating heat shrink tube is disposed on an outer surface of the reference electrode.
3. The blood glucose sensor of claim 1, wherein the working electrode is a platinum iridium working electrode and the insulating layer on the outer surface of the working electrode is a polytetrafluoroethylene layer.
4. The blood glucose sensor of claim 1, wherein the wireless transmission device comprises an electrochemical signal amplifier, a wireless transmission module and an on-board antenna, wherein the electrochemical signal amplifier is electrically connected with the working electrode and the reference electrode through electrode leads respectively; the wireless transmission module is electrically connected with the electrochemical signal amplifier; the onboard antenna is electrically connected with the wireless transmission module.
5. The blood glucose sensor of claim 1, wherein the epoxy layer has a diameter of 0.175mm and a length of 0.3 mm.
6. The blood glucose sensor of claim 1, wherein the length of the perfluorosulfonic acid polymer membrane is 1 mm.
7. A preparation method of a blood glucose sensor is characterized by comprising the following steps:
adopting platinum-iridium alloy materials to manufacture a working electrode, setting one end part of the manufactured working electrode as an induction cavity, and covering a polytetrafluoroethylene layer on the surface of the working electrode between the other end part of the working electrode and the induction cavity;
winding copper wires on the polytetrafluoroethylene layer area of the working electrode in an interval spiral mode, and covering the copper wires with a silver/silver chloride slurry layer to form a reference electrode;
coating an epoxy resin layer on the outer end of the induction cavity, coating 5-6% of perfluorosulfonic acid polymer solution on the surface of the rest area of the induction cavity between the outer end of the induction cavity and the polytetrafluoroethylene layer, and air-drying to form a perfluorosulfonic acid polymer film; the diameter of the epoxy resin layer is 0.175mm-0.2mm, and the length is 0.3mm-0.4 mm; the length of the perfluorinated sulfonic acid polymer membrane is 1mm-1.1 mm;
soaking the working electrode coated with the perfluorosulfonic acid polymer membrane in 0.25-3ml of phosphate buffer solution containing 2.5-3mg of glucose oxidase and dissolved with 5-7mg of bovine serum albumin, wherein the concentration of the phosphate buffer solution is 0.1mol/L, drying in the air for 10-15 minutes, and repeating the process at least twice;
immersing the working electrode induction cavity into 25-30% glutaraldehyde solution for 5-6 h;
an electrochemical signal amplifier is respectively and electrically connected with the working electrode and the reference electrode through electrode leads;
electrically connecting a wireless transmission module with the electrochemical signal amplifier;
and electrically connecting an onboard antenna with the wireless transmission module.
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| CN201510680579.5A CN105361891A (en) | 2015-10-19 | 2015-10-19 | Blood glucose sensor planted in live fish eye ball sclera interstitial fluid and production method of blood glucose sensor |
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| WO2023246519A1 (en) * | 2022-06-22 | 2023-12-28 | 苏州百孝医疗科技有限公司 | Implantable electrochemical biosensor, test method, and implantable medical instrument |
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| CN106706726B (en) * | 2016-12-09 | 2019-08-23 | 中国农业大学 | Blood sugar monitoring methods and device in a kind of transport of aquatic products waterless keep-alive |
| CN111870238A (en) * | 2019-05-03 | 2020-11-03 | 乌宁 | Implanted biosensor and manufacturing method thereof |
| WO2021031057A1 (en) * | 2019-08-19 | 2021-02-25 | Medtrum Technologies Inc. | Sensing device |
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| US5165407A (en) * | 1990-04-19 | 1992-11-24 | The University Of Kansas | Implantable glucose sensor |
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| CN103687958B (en) * | 2011-04-18 | 2015-11-25 | 诺威奥森斯有限公司 | Biosensor |
| WO2014041190A1 (en) * | 2012-09-17 | 2014-03-20 | Brains Online Holding B.V. | Rod shaped implantable biosensor |
| WO2014110492A2 (en) * | 2013-01-11 | 2014-07-17 | Northeastern University | Saliva glucose monitoring system |
| CN103462615B (en) * | 2013-09-13 | 2015-12-16 | 上海移宇科技有限公司 | Micrometer-scale glucose sensor microelectrode |
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- 2015-10-19 CN CN202110101847.9A patent/CN112932477A/en active Pending
- 2015-10-19 CN CN201510680579.5A patent/CN105361891A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US5165407A (en) * | 1990-04-19 | 1992-11-24 | The University Of Kansas | Implantable glucose sensor |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023246519A1 (en) * | 2022-06-22 | 2023-12-28 | 苏州百孝医疗科技有限公司 | Implantable electrochemical biosensor, test method, and implantable medical instrument |
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