CN116087114A - Optical fiber biosensor based on electrostatic spinning technology and biological detection method - Google Patents
Optical fiber biosensor based on electrostatic spinning technology and biological detection method Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
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Abstract
The invention discloses an optical fiber biosensor based on an electrostatic spinning technology and a biological detection method; the optical fiber biosensor comprises an optical fiber and a nanofiber; wherein, part of the fiber core of the optical fiber is exposed; the nanofiber is obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber; the raw materials are obtained by mixing a biosensing material and an electrostatic spinning material; when the optical fiber biosensor works, the optical fiber outputs an optical signal representing the concentration of a target component in a measured sample; the biosensing material is a material capable of generating a biochemical reaction with the target component. The biological sensitive material in the optical fiber biosensor provided by the invention is not easy to fall off, the activity is not reduced, and the optical fiber biosensor has higher sensing performance.
Description
Technical Field
The invention belongs to the field of biosensors, and particularly relates to an optical fiber biosensor based on an electrostatic spinning technology and a biological detection method.
Background
A biosensor is a device sensitive to a specific biological substance, capable of converting the concentration of the biological substance into an electrical signal or an optical signal to perform measurement of biological components; the characteristic that a biosensor is sensitive to a specific biological substance is achieved by loading the sensor with a material sensitive to the substance, which is called a biosensing material. For example, in biosensors manufactured based on enzyme immobilization technology, enzymes are a highly efficient biocatalyst as a biosensing material, and the reaction rate occurring in the biosensor can be generally increased by 3 to 20 orders of magnitude under the catalysis of enzymes.
In the actual manufacturing of biosensors based on enzyme immobilization technology, the immobilization of enzymes on the biosensor is a first concern, since enzyme immobilization is often a major factor affecting the performance of the biosensor.
In the prior art, enzyme immobilization techniques exist in various ways, including a cross-linking method enzyme immobilization technique, a covalent bonding enzyme immobilization technique, an adsorption method enzyme immobilization technique, and the like. Wherein, the crosslinking method enzyme immobilization technology utilizes the crosslinking action between the difunctional or multifunctional crosslinking agent and enzyme molecules to generate a network structure so as to realize the action of immobilized enzyme. However, the cross-linking process between the cross-linker and the enzyme molecule tends to cause the enzyme to lose activity, thereby affecting the performance of the biosensor. The covalent bonding enzyme immobilization technology is to react the active functional groups on the surface of the carrier with the active nonessential groups of the enzyme to form covalent bonds, so as to achieve the purpose of enzyme immobilization. However, due to the presence of covalent bonds, the structure of the enzyme is changed and the activity of the enzyme is also reduced, thereby affecting the performance of the biosensor. The adsorption enzyme immobilization technique is a method of adsorbing and immobilizing an enzyme on a surface by using various adsorbents. The method utilizes the interaction force of enzyme molecules and polar bonds, hydrophobic bonds, ionic bonds, van der Waals forces and hydrogen bonds of a carrier to adsorb and fix the enzyme. However, the enzyme-carrier interaction in the adsorption enzyme immobilization technique is not strong, and the enzyme is easily released.
In view of the foregoing, there is a need in the art for a high performance biosensor that is not only resistant to the release of the loaded biosensing material during operation of the biosensor, but also does not degrade the inherent sensitivity characteristics of the biosensing material.
Disclosure of Invention
In order to solve the technical problems in the prior art, the invention provides an optical fiber biosensor based on an electrostatic spinning technology and a biological detection method.
The technical problems to be solved by the invention are realized by the following technical scheme:
an optical fiber biosensor based on an electrostatic spinning technology comprises an optical fiber and a nanofiber; wherein,,
part of the fiber core of the optical fiber is exposed;
the nanofiber is obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber; the raw materials are obtained by mixing a biosensing material and an electrostatic spinning material;
when the optical fiber biosensor works, the optical fiber outputs an optical signal representing the concentration of a target component in a measured sample; the biosensing material is a material capable of generating a biochemical reaction with the target component.
Optionally, the bare fiber core is partially polished.
Optionally, the polished portion of the bare fiber core is semi-cylindrical.
Optionally, the polished portion of the bare core is tapered or tapered with a pair of vertices offset.
Optionally, the biosensing material comprises: an enzyme, an antibody, an antigen or a nucleic acid.
Optionally, the optical fiber biosensor includes: a fiber optic biosensor for detecting glucose;
the biosensing material comprises: glucose hexokinase;
the electrospun material comprises: polyvinyl alcohol;
the nanofiber is specifically obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber and then carrying out waterproof treatment on an electrostatic spinning product.
Optionally, the waterproofing treatment includes: the electrospun product is crosslinked with glutaraldehyde vapor.
The embodiment of the invention also provides a biological detection method based on the electrostatic spinning technology, which comprises the following steps:
placing any optical fiber biosensor based on the electrostatic spinning technology into a sample to be tested;
inputting light waves into an optical fiber of the optical fiber biosensor by using a broadband light source;
detecting the light waves output from the optical fiber by using a spectrometer to obtain a detection spectrum;
determining the concentration of the target component in the sample to be tested based on the detection spectrum.
Optionally, the inputting light waves into the optical fiber of the optical fiber biosensor by using a broadband light source includes: enabling light waves emitted by a broadband light source to enter an input port of an optical circulator and enter an optical fiber of the optical fiber biosensor from an output port of the optical circulator;
the detecting the light wave output from the optical fiber by using a spectrometer comprises: light waves reflected from the optical fiber are detected from a feedback port of the optical circulator using a spectrometer.
Optionally, the inputting light waves into the optical fiber of the optical fiber biosensor by using a broadband light source includes: inputting light waves to one end of an optical fiber of the optical fiber biosensor by using a broadband light source;
the detecting the light wave output from the optical fiber by using a spectrometer comprises: the light waves transmitted from the optical fiber are detected from the other end of the optical fiber using a spectrometer.
In the optical fiber biosensor based on the electrostatic spinning technology, the electrostatic spinning technology is adopted to fix the biosensing material in the nanofiber; because the electrostatic spinning is a physical process, the inherent biochemical characteristics of the biosensing material are not affected, so that the inherent sensitivity characteristics of the biosensing material can be maintained; and as the biosensing material is mixed in the raw material of the electrostatic spinning, but not attached to the electrostatic spinning, the biosensing material is not easy to fall off. In addition, the diameter of the nanofiber is nanoscale, and the biosensing material mixed in the nanofiber can still be fully contacted and reacted with an external measured sample, so that the optical fiber biosensor provided by the invention has higher sensing performance.
The present invention will be described in further detail with reference to the accompanying drawings.
Drawings
FIG. 1 is a schematic structural diagram of an optical fiber biosensor based on an electrostatic spinning technology according to an embodiment of the present invention;
figures 2 and 3 show the microstructure of the nanofibers at different resolutions, respectively;
FIG. 4 is a schematic diagram of an electrospinning apparatus used in the process of preparing the optical fiber biosensor shown in FIG. 1;
FIGS. 5 to 7 are schematic structural views of three other optical fiber biosensors based on the electrospinning technology according to the embodiment of the present invention;
FIG. 8 is a FT-IR spectrum of a polyvinyl alcohol not crosslinked with glutaraldehyde and a FT-IR spectrum of a polyvinyl alcohol after crosslinking with glutaraldehyde;
FIG. 9 is a schematic flow chart of a biological detection method based on an electrostatic spinning technology according to an embodiment of the present invention;
FIG. 10 is a schematic diagram of a test environment for use in performing the method of FIG. 9;
FIG. 11 is a schematic diagram of another testing environment used in performing the method of FIG. 9; .
Detailed Description
The present invention will be described in further detail with reference to specific examples, but embodiments of the present invention are not limited thereto.
In order to solve the problems that a biosensing material of a biosensor is easy to fall off and the activity is easy to reduce in the prior art, the embodiment of the invention provides an optical fiber biosensor based on an electrostatic spinning technology. Referring to fig. 1, the optical fiber biosensor includes: optical fibers and nanofibers. Wherein, part of the fiber core of the optical fiber is exposed; the nanofiber is obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber; fig. 2 and 3 show the microstructure of the nanofibers at different resolutions, respectively. Wherein, the raw materials used for preparing the nanofiber are obtained by mixing a biosensing material and an electrostatic spinning material. It is understood that a biosensing material is a material that is capable of biochemically reacting with and is sensitive to a target component in a sample being tested.
When the optical fiber biosensor works, the optical fiber outputs an optical signal representing the concentration of a target component in a measured sample. Specifically, light waves are input into the optical fiber, and the light waves contact the nanofiber through the exposed fiber core of the optical fiber; the biological sensitive material contained in the nanofiber reacts with the target component in the sample to be tested, so that the refractive index of the light wave at the exposed fiber core is changed. Therefore, by detecting the light waves reflected or transmitted by the optical fiber, the concentration of the target component corresponding to the refractive index of the current sensor of the optical fiber can be determined by performing spectral analysis.
The above-mentioned bio-sensitive material may include: an enzyme, an antibody, an antigen or a nucleic acid.
The above-mentioned electrospun material may include: polypropylene, cellulose acetate, l-polylactic acid, polyethylene oxide, polyurethane, polyamide, polyvinyl alcohol, or the like. In the specific preparation of the electrospun raw material, it may be sufficiently dissolved with a solvent suitable for the electrospun material to be mixed with the biosensing material. The solvent suitable for the electrospinning material is a material which can dissolve the electrospinning material and does not affect the properties of the electrospinning material itself. It is known to those skilled in the art that some electrospun materials are water soluble, while some are oil soluble, and that other electrospun materials are both water soluble and oil soluble, so the choice of solvent needs to be specifically determined based on the electrospun material. For example, when polyvinyl alcohol is used as an electrospinning material, the polyvinyl alcohol can be dissolved in PBS (phosphate buffered saline, phosphate buffer) and sufficiently dissolved by stirring with a magnetic stirrer. For another example, when cellulose acetate is used as an electrospinning material, cellulose acetate may be dissolved in an acetone solution, a methylene chloride solution or a DMF (dimethylformamide) solution to be sufficiently dissolved.
And fully mixing the dissolved electrostatic spinning material with the biosensing material to obtain the electrostatic spinning raw material. And then, carrying out electrostatic spinning on the raw materials to the exposed fiber core of the optical fiber by using an electrostatic spinning device, so as to obtain the optical fiber biosensor provided by the embodiment of the invention.
FIG. 4 illustrates a simple electrospinning apparatus; the micropump is responsible for pushing the mixed solution of the electrostatic spinning material and the biological sensitive material so as to enable the mixed solution to be sprayed out of the needle head uniformly and stably; the optical fiber is placed on the collection plate with the bare fiber core aligned with the needle; under the action of static electricity released by a power supply, the mixed solution is collected at the exposed fiber core of the optical fiber and is tightly attached to the fiber core.
When the electrospinning device shown in fig. 4 is used for electrospinning, the advancing speed of the mixed solution may be 0.0015mm/s, the voltage of the power supply for electrostatic discharge may be 18.5kV, the distance between the needle and the collecting plate may be 12cm, and the specification of the needle may be 18G. The time to be electrospun may be set to four minutes.
It should be noted that, in practical applications, the advancing speed of the mixed solution, the voltage of the electrostatic discharge, the distance between the needle and the collecting plate, and the specification of the needle are not limited to the above-listed examples, and may be adjusted according to the actual spinning requirements and the experimental spinning effect. For example, the thickness of the nanofibers can be controlled by adjusting the gauge of the needle; the specification of the needle can be selected from 8G to 30G.
In practice, it is generally first determined which substance the optical fiber biosensor to be prepared is specifically used for detecting, and thus the biologically sensitive material sensitive to the substance is correspondingly determined. Then, an electrostatic spinning material which is not easy to generate biochemical reaction with the biological sensitive material is selected to further manufacture raw materials required by electrostatic spinning.
In the optical fiber biosensor based on the electrostatic spinning technology provided by the embodiment of the invention, the electrostatic spinning technology is adopted to fix the biosensing material in the nanofiber; because the electrostatic spinning is a physical process, the inherent biochemical characteristics of the biosensing material are not affected, so that the inherent sensitivity characteristics of the biosensing material can be maintained; and as the biosensing material is mixed in the raw material of the electrostatic spinning, but not attached to the electrostatic spinning, the biosensing material is not easy to fall off. In addition, the diameter of the nanofiber is nanoscale, and the biosensing material mixed in the nanofiber can still be fully contacted and reacted with an external measured sample, so that the optical fiber biosensor provided by the embodiment of the invention has higher sensing performance.
In one embodiment, to increase the amount of nanofibers, the optical fiber biosensor has a higher detection performance, and the nanofibers are more tightly attached to the bare fiber core, which may be further polished. For example, the bare cores are polished to form the structure shown in fig. 5-7. The polished portion of the bare core in fig. 5 is semi-cylindrical; the polished portion of the bare core in fig. 6 is tapered; the polished portion of the bare core in fig. 7 is tapered with a pair of vertices against each other. In this way, the polished area of the fiber core can contain more nanofibers, and the nanofibers are embedded in the polished area and are tightly attached to the fiber core.
Comparing fig. 5 to 7 with fig. 1, it can be seen that the optical fiber biosensor obtained in fig. 5 to 7 has a significantly increased amount of nanofibers compared with fig. 1, so that the amount of the bio-sensitive material in the optical fiber biosensor can be increased, thereby enabling the optical fiber biosensor to have higher detection performance.
It is understood that the polishing method of the embodiment of the present invention for polishing the bare fiber core is not limited to the polishing method shown in fig. 5 to 7, and any polishing method that can increase the amount of nanofibers and make the optical fiber biosensor have a higher detection performance may be applied to the embodiment of the present invention.
In the following, an optical fiber biosensor based on the electrospinning technology provided by the embodiment of the present invention will be described in more detail by taking a glucose biosensor occupying 85% of the entire biosensor market as an example.
Specifically, in order to be able to effectively detect glucose, glucose hexokinase sensitive to glucose is selected as a biosensing material in the optical fiber biosensor. A number of materials such as polyvinyl alcohol, polymethyl methacrylate, chitosan, etc. are known to be useful as substrates for enzyme immobilization in glucose biosensors. It is assumed here that polyvinyl alcohol is selected as the electrospinning material.
8g of polyvinyl alcohol was added to 100ml of PBS and stirred by a magnetic stirrer for 6 hours until the polyvinyl alcohol was completely dissolved in PBS, to obtain a polyvinyl alcohol buffer solution. 10ml of a polyvinyl alcohol buffer solution was taken, 20mg of glucohexokinase powder was added thereto, and magnetic stirring was performed for 1 hour until glucohexokinase was uniformly dispersed in the polyvinyl alcohol buffer solution, and the resulting mixed solution was a raw material for preparing nanofibers.
Then, based on the obtained raw material, electrospinning was performed on the polished optical fiber shown in fig. 5, 6, or 7 by using an electrospinning device, to form the structure shown in the figure.
Since polyvinyl alcohol is water-soluble, the electrospun product, i.e., the freshly spun fibers, can be further subjected to a water-repellent treatment at this time. Specifically, at room temperature, the optical fiber together with the spun fiber was placed in a sealed container containing 0.5 to 2.56 mol of a glutaraldehyde solution having a volume ratio of 10%, and 1ml of HCl (hydrochloric acid) was added as a catalyst. Thus, glutaraldehyde volatilizes in the sealed container, and the generated gas and the fiber undergo a crosslinking reaction; after 24 hours, taking out the optical fiber and the fiber together, putting the optical fiber and the fiber into a drying box for drying for 24 hours, and ensuring that glutaraldehyde vapor which is not fully reacted on the optical fiber and the fiber is completely volatilized; thus, a prepared glucose biosensor was obtained.
FIG. 8 shows the FT-IR spectrum of polyvinyl alcohol not crosslinked with glutaraldehyde and the FT-IR spectrum of polyvinyl alcohol after crosslinking with glutaraldehyde; the horizontal axis represents the wave number of the infrared light wave, and the vertical axis represents the light transmission rate. From FIG. 8, it can be seen that the characteristic peak at 3327cm-1 belongs to the stretching of-OH. After crosslinking of the polyvinyl alcohol with glutaraldehyde, stretching of C=O at 1734cm-1 and stretching of acetal bonds (-C-O-C-) near 1089cm-1 can be observed from FIG. 8, and it can also be found that the strength of the-OH absorption peak at 3329cm-1 decreases after crosslinking, which both indicate that the polyvinyl alcohol has been water insoluble. The reason for the modification of polyvinyl alcohol after crosslinking is that the-OH concentration in the polyvinyl alcohol is reduced due to the crosslinking reaction between polyvinyl alcohol and glutaraldehyde.
Corresponding to the optical fiber biosensor based on the electrostatic spinning technology, the embodiment of the invention also provides a biological detection method based on the electrostatic spinning technology, as shown in fig. 9, the method comprises the following steps:
s10: and placing the optical fiber biosensor based on the electrostatic spinning technology into a sample to be tested.
S20: the light waves are input into the optical fibers of the optical fiber biosensor by using a broadband light source.
S30: and detecting the light waves output from the optical fiber by using a spectrometer to obtain a detection spectrum.
S40: the concentration of the target component in the sample to be measured is determined based on the detection spectrum.
The optical fiber biosensor based on the electrostatic spinning technology used in step S10 may be any of the above optical fiber biosensors based on the electrostatic spinning technology.
In one implementation, referring to fig. 10, the inputting of the light wave into the optical fiber of the optical fiber biosensor using the broadband light source in step S20 may include: the light wave emitted by the broadband light source enters an input port of an optical circulator and enters an optical fiber of the optical fiber biosensor from an output port of the optical circulator. Accordingly, detecting the light wave output from the optical fiber using the spectrometer in step S30 includes: the light waves reflected from the optical fiber are detected from a feedback port of the optical circulator using a spectrometer.
In another implementation, referring to fig. 11, inputting the light wave into the optical fiber of the optical fiber biosensor using the broadband light source in step S20 may include: the light wave is input to one end of the optical fiber biosensor by using a broadband light source. Accordingly, detecting the light wave output from the optical fiber using the spectrometer in step S30 includes: the light waves transmitted from the optical fiber are detected from the other end of the optical fiber using a spectrometer.
Determining the concentration of the target component in the sample to be measured based on the detection spectrum in step S40 includes: and determining the concentration corresponding to the currently obtained detection spectrum according to the corresponding relation between the plurality of groups of detection spectrums and the concentration of the target component, namely the concentration of the target component in the detected sample.
It should be noted that, for the method embodiment, since it is substantially similar to the product embodiment, the description is relatively simple, and the relevant points are referred to in the section of the product embodiment.
In the description of the present specification, the meaning of "a plurality" is two or more, unless explicitly defined otherwise. The description of the terms "one embodiment," "some embodiments," "example," "specific examples," or "some examples," etc., means that a particular feature or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Further, one skilled in the art can engage and combine the different embodiments or examples described in this specification.
Although the present application has been described herein in connection with various embodiments, other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed application, from a review of the figures, the disclosure, and the appended claims.
The foregoing is a further detailed description of the invention in connection with the preferred embodiments, and it is not intended that the invention be limited to the specific embodiments described. It will be apparent to those skilled in the art that several simple deductions or substitutions may be made without departing from the spirit of the invention, and these should be considered to be within the scope of the invention.
Claims (10)
1. An optical fiber biosensor based on an electrostatic spinning technology is characterized by comprising an optical fiber and a nanofiber; wherein,,
part of the fiber core of the optical fiber is exposed;
the nanofiber is obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber; the raw materials are obtained by mixing a biosensing material and an electrostatic spinning material;
when the optical fiber biosensor works, the optical fiber outputs an optical signal representing the concentration of a target component in a measured sample; the biosensing material is a material capable of generating a biochemical reaction with the target component.
2. The fiber optic biosensor according to claim 1, wherein the bare fiber core is partially polished.
3. The fiber optic biosensor according to claim 2, wherein the polished portion of the bare fiber core is semi-cylindrical.
4. The fiber optic biosensor according to claim 2, wherein the polished portion of the bare fiber core is tapered or tapered with a pair of vertices offset.
5. The fiber optic biosensor according to claim 1, wherein the biosensing material comprises: an enzyme, an antibody, an antigen or a nucleic acid.
6. The fiber optic biosensor of claim 1, wherein the fiber optic biosensor comprises: a fiber optic biosensor for detecting glucose;
the biosensing material comprises: glucose hexokinase;
the electrospun material comprises: polyvinyl alcohol;
the nanofiber is specifically obtained by carrying out electrostatic spinning on raw materials to the exposed fiber core of the optical fiber and then carrying out waterproof treatment on an electrostatic spinning product.
7. The fiber optic biosensor according to claim 6, wherein the waterproofing treatment comprises: the electrospun product is crosslinked with glutaraldehyde vapor.
8. The biological detection method based on the electrostatic spinning technology is characterized by comprising the following steps of:
placing the optical fiber biosensor based on the electrostatic spinning technology according to any one of claims 1-7 into a sample to be tested;
inputting light waves into an optical fiber of the optical fiber biosensor by using a broadband light source;
detecting the light waves output from the optical fiber by using a spectrometer to obtain a detection spectrum;
determining the concentration of the target component in the sample to be tested based on the detection spectrum.
9. The method according to claim 8, wherein,
the inputting light waves into the optical fiber of the optical fiber biosensor by using a broadband light source comprises: enabling light waves emitted by a broadband light source to enter an input port of an optical circulator and enter an optical fiber of the optical fiber biosensor from an output port of the optical circulator;
the detecting the light wave output from the optical fiber by using a spectrometer comprises: light waves reflected from the optical fiber are detected from a feedback port of the optical circulator using a spectrometer.
10. The method according to claim 8, wherein,
the inputting light waves into the optical fiber of the optical fiber biosensor by using a broadband light source comprises: inputting light waves to one end of an optical fiber of the optical fiber biosensor by using a broadband light source;
the detecting the light wave output from the optical fiber by using a spectrometer comprises: the light waves transmitted from the optical fiber are detected from the other end of the optical fiber using a spectrometer.
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