CN105842208A - Beta-dextran detection method based on fluorescence enhancement principle - Google Patents
Beta-dextran detection method based on fluorescence enhancement principle 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
Abstract
The invention discloses a beta-dextran detection method based on a fluorescence enhancement principle. The beta-dextran detection method comprises the steps: with a treated silicon slice as a substrate, combining with a spin-coating technique, and depositing titanium dioxide and graphene oxide hydrogel on the silicon slice, to prepare titanium dioxide/graphene oxide hydrogel one-dimensional photonic crystals; plating the photonic crystals with gold by means of a vacuum evaporation plating method, to prepare gold-plated titanium dioxide/graphene oxide hydrogel one-dimensional photonic crystals; taking the photonic crystals as a fluorescence enhancement substrate, and detecting beta-dextran through a method of detecting the fluorescence intensity of a fluorescence composite product formed by specifically combining beta-dextran and aniline blue. The gold-plated titanium dioxide/graphene oxide hydrogel one-dimensional photonic crystals are used as the substrate, fluorescence enhancement effects of gold nanoparticles and the photonic crystals is combined, and more substantial fluorescence enhancement is caused, so as to improve the sensitivity of the fluorescence detection. The method is simple, effective, and simple and convenient to operate, and in required time is relatively short.
Description
Technical field
The present invention relates to Fluorescence Increasing technical field, be specifically related to gold-plated titanium dioxide/one-dimensional light of graphene oxide hydrogel
The preparation method of sub-crystal.
Background technology
Light photonic crystal can be seen as the regular texture that the material periodicities arrangement of differing dielectric constant is formed.
Yablonovitch and John studies at first in terms of photonic crystal designs.According to periodically, photonic crystal is permissible
It is divided into one-dimensional, two dimension, three-D photon crystal.Photonic crystal has different transmitting wavelength, this be by between layers away from
From determine, this also form the color of they uniquenesses.When the periodicity of photonic crystal changes when, such as, meet
Having arrived chemical stimulation, the reflection wavelength of photonic crystal also can change.This just provides a convenient approach for sensing.
Even if being that unfamiliar people can also be observed that the color of photonic crystal changes, and this method being cheap, easily operates.
This makes to contrast other stimuli responsive shaped materials, and photonic crystal is more welcome.
1-D photon crystal can be not only used for sensing, and tool research simultaneously shows that 1-D photon crystal has certain Fluorescence Increasing
Effect, can be used for fluoroscopic examination.Owing to the existence of forbidden photon band makes photonic crystal have special optical characteristics, photon
Crystal can affect transmitting and the propagation of light.Especially forbidden photon band can suppress the light biography in a direction of characteristic frequency
Broadcast, therefore brilliant with photon when the transmitting wavelength of the optical material (dyestuff, polymer semiconductor etc.) embedded in photonic crystal
When the forbidden photon band of body coincides, photonic crystal can change the emission characteristics of optical material.1-D photon crystal strengthens glimmering
One mechanism of light is as follows: when the wavelength of light coincides with the forbidden photon band of 1-D photon crystal, light is just by one-dimensional photon
Crystal is suppressed, say, that launches light and cannot be introduced into 1-D photon crystal.So light can be anti-by the direction towards forbidden photon band
Penetrate.But when light incide do not have on the sheet glass of 1-D photon crystal time, due to the antiradar reflectivity of sheet glass, light can quilt
Travel in substrate.This has indicated that light source is transmitted into when the wavelength of light overlaps with the forbidden photon band of 1-D photon crystal
The light on 1-D photon crystal surface will be propagated through 1-D photon crystal, but is reflected to the direction of forbidden photon band
And it is enhanced.And when substrate is sheet glass when, light can be transmitted and scattered and disappeared.
The surface (such as metal surface) with special appearance and configuration can make to be positioned at the glimmering of the neighbouring fluorescence molecule in its surface
Optical signal is obviously enhanced, and this phenomenon is referred to as surface-enhanced fluorescence.It is many factors effect that metal surface strengthens fluorescence
As a result, the theory of relevant mechanism and enhancement mechanism mainly has three: the enhancing of local fields near the fluorescence molecule of (1) metal surface: gold
The plasma that metal surface is formed can make excitation intensity and the efficiency of fluorescence molecule by local electric field near fluorescence molecule
Strengthen.(2) the energy transfer between molecule and substrate surface: according to Forster-Dexter mechanism, metal surface fluorescence divides
The enhancing of sub-quantum yield is also understood to be that fluorescence molecule is quickly transferred to metal surface energy and is radiated the knot in far field
Really.(3) increase of fluorescence molecule attenuation speed: metal can be by resonance coupling strong between photon, greatly
Ground increases the electromagnetic field of metallic perimeter, thus increases substantially the radiative transistion probability of the centre of luminescence, realizes Fluorescence Increasing with this.
These three theory explains that metal surface strengthens the reason of fluorescence the most from different perspectives, but is to be mutually related.
Without researcher, with metal surface, the Fluorescence Increasing effect of 1-D photon crystal is strengthened fluorescence the most temporarily to combine, and profit
Fluorescence Increasing effect after combining with it detects guest molecule.Therefore, the Fluorescence Increasing of 1-D photon crystal is imitated by research
The experiment that fruit combines with metal surface enhancing fluorescence needs more deep.
Summary of the invention
Technical problem: it is an object of the invention to provide a kind of beta glucan detection method based on Fluorescence Increasing principle, fill out
Mend and the Fluorescence Increasing effect of 1-D photon crystal is combined with metal surface enhancing fluorescence, and utilize the fluorescence after its combination
Reinforced effects detects the blank in terms of guest molecule.
Technical scheme: for solving the problems referred to above, a kind of based on Fluorescence Increasing principle the beta glucan detection side of the present invention
Method is by the following technical solutions:
In conjunction with the Fluorescence Increasing effect of golden nanometer particle and photonic crystal, comprise the steps:
Step one, prepare TiO respectively2Colloid solution and GO hydrogel, wherein, TiO2Colloid solution concentration is 7.8～39
Mg/ml, GO hydrogel concentration is 0.25～0.5mg/ml；
Step 2, silicon chip are cleaned and are dried up, and utilize the TiO that step one is prepared by spin-coating method2Colloid solution and GO hydrogel are coated with
To silicon chip surface, obtain described titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, every layer of spin coating rotating speed
Being 3000～6000rpm, spin-coating time is 50s～1min；
Step 3, by the method for vacuum evaporation on titanium dioxide/graphene oxide hydrogel 1-D photon crystal gold-plated,
Obtaining described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, the evaporation time is 60s～90s；
Step 4, beta glucan is specific binding with aniline blue, obtain fluorescence combination product, wherein, beta glucan
Solution concentration is 1～6 μ g/ml, and aniline blue solution concentration is 0.5～1.5mg/ml；
Step 5, the fluorescence combination product obtained by step 4 is mixed in proportion with polyvinyl alcohol, utilizes spin-coating method by upper
State solution and be coated onto gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal surface, obtain described with gold-plated two
Titanium oxide/graphene oxide hydrogel 1-D photon crystal is the fluorescence combination product of Fluorescence Increasing substrate, wherein, polyethylene
Alcoholic solution concentration is 20～40mg/ml, and mixed proportion is 1: 1～2: 1, and spin coating rotating speed is 900～1200rpm, during spin coating
Between be 10s～12s；
Step 6, utilize prepared by fluorescence microscope detecting step five with gold-plated titanium dioxide/graphene oxide hydrogel one
Dimensional photonic crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
Described TiO2The preparation of colloid solution comprises the steps: that weighing butyl titanate adds in ethanol, magnetic agitation
Under be slowly added dropwise glacial acetic acid, wherein butyl titanate, the volume ratio of ethanol and glacial acetic acid is 2: 8: 1, under room temperature stir 5～6
H, obtains the TiO that concentration is 39mg/ml2Colloid solution, dilutes 1～5 times, and obtaining concentration is 7.8～39mg/ml
TiO2Colloid solution.
The preparation of described GO hydrogel comprises the steps: to fall the GO hydrogel of prepare 0.25～0.5mg/mL
Enter in autoclave, 90～100 DEG C of reactions 17～24h, obtain the GO hydrogel that concentration is 0.25～0.5mg/ml.
Described silicon chip is cleaned to dry up and is comprised the steps: silicon chip to immerse acetone soln sonic oscillation 30～60min, afterwards
Immerse sonic oscillation 30～60min in ethanol solution again；It is finally immersed in the H that volume ratio is 3: 7～1: 32SO4/H2O2Mixing
Solution soaks 4～12h, N2Dry up the silicon chip obtaining cleaning up.
To be coated with one layer of TiO2Colloid solution and one layer of GO hydrogel are a spin coating cycle, and the described spin coating cycle is 3～4
The individual cycle.
With aniline blue is specific binding, beta glucan comprises the steps: that taking beta glucan solution puts in centrifuge tube,
Adding aniline blue solution in centrifuge tube, wherein the volume ratio of beta glucan solution and aniline blue solution is 1: 21, lucifuge
50～80 DEG C of water-baths 15～30min, obtain the fluorescence combination product of aniline blue and the specific binding formation of beta glucan.
Beneficial effect:
1, the present invention is with gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal as substrate, by combining gold
The Fluorescence Increasing effect of nanoparticle and photonic crystal, causes Fluorescence Increasing by a larger margin, thus promotes fluoroscopic examination
Sensitivity, thus the gold-plated titanium dioxide prepared/graphene oxide hydrogel 1-D photon crystal is than common one-dimensional photon
Crystal film Fluorescence Increasing ability is higher.
2, the inventive method is simply effective, easy and simple to handle, and required time is shorter.
3, the preparation facilities that the present invention uses is simple: need not what special equipment, common spin coater just can be expired
Foot requirement.
4, gold-plated titanium dioxide prepared by the present invention/graphene oxide hydrogel 1-D photon crystal applied range: system
The inspection of multiple guest molecule fluorescence is may be used for for the gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal gone out
Fluorescence Increasing substrate in survey, is with a wide range of applications.
Accompanying drawing explanation
Fig. 1 is that the fluorescence of gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal that embodiment 1 is prepared increases
Potent fruit.
Fig. 2 is that gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal of preparing of embodiment 1 is for examining
Survey the result figure of beta glucan.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing the present invention done and further explain.The following example is merely to illustrate this
Bright, but it is not used to limit the practical range of the present invention.
A kind of beta glucan detection method based on Fluorescence Increasing principle, in conjunction with golden nanometer particle and the fluorescence of photonic crystal
Reinforced effects, comprises the steps:
Step one, silicon chip is immersed acetone soln sonic oscillation 30～60min, immerse Under Ultrasonic Vibration in ethanol solution the most again
Swing 30～60min；It is finally immersed in the H that volume ratio is 3: 7～1: 32SO4/H2O2Mixed solution soaks 4～12h, N2Blow
The dry silicon chip obtaining cleaning up；
Step 2, prepare TiO respectively2Colloid solution and GO hydrogel, wherein, TiO2Colloid solution concentration is 7.8～39
Mg/ml, GO hydrogel concentration is 0.1～0.5mg/ml；Wherein, TiO2The preparation of colloid solution comprises the steps:
Weigh butyl titanate to add in ethanol, be slowly added dropwise glacial acetic acid under magnetic agitation, wherein butyl titanate, ethanol and ice
The volume ratio of acetic acid is 2: 8: 1, stirs 5～6h, obtain the TiO that concentration is 39mg/ml under room temperature2Colloid solution is dilute
Release 1～5 times, obtain the TiO that concentration is 7.8～39mg/ml2Colloid solution.The preparation of GO hydrogel includes walking as follows
Rapid: the GO hydrogel of prepare 0.25～0.5mg/mL is poured in autoclave, 90～100 DEG C of reactions 17～24
H, obtains the GO hydrogel that concentration is 0.25～0.5mg/ml.
Step 3, utilize the TiO that step 2 prepared by spin-coating method2Colloid solution and GO hydrogel are coated onto silicon chip surface,
To described titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, every layer of spin coating rotating speed is 3000～6000
Rpm, spin-coating time is 50s～1min, to be coated with one layer of TiO2Colloid solution and one layer of graphene oxide colloid solution are one
In the individual spin coating cycle, the spin coating cycle is 3～4 cycles.
Step 4, by the method for vacuum evaporation on titanium dioxide/graphene oxide hydrogel 1-D photon crystal gold-plated,
Obtaining described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, the evaporation time is 60s～90s.
Step 5, beta glucan is specific binding with aniline blue, obtain fluorescence combination product, wherein, beta glucan
Solution concentration is 1～6 μ g/ml, and aniline blue solution concentration is 0.5～1.5mg/ml, wherein, and beta glucan and aniline blue
Specific binding comprise the steps: that taking a certain amount of beta glucan solution puts in centrifuge tube, add in centrifuge tube
Aniline blue solution, wherein the volume ratio of beta glucan solution and aniline blue solution is 1: 21, lucifuge 50～80 DEG C of water-baths
15～30min, obtain the fluorescence combination product of aniline blue and the specific binding formation of beta glucan.
Step 6, the fluorescence combination product obtained by step 5 is mixed in proportion with polyvinyl alcohol, utilizes spin-coating method by upper
State solution and be coated onto gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal surface, obtain described with gold-plated two
Titanium oxide/graphene oxide hydrogel 1-D photon crystal is the fluorescence combination product of Fluorescence Increasing substrate, wherein, polyethylene
Alcoholic solution concentration is 20～40mg/ml, and mixed proportion is 1: 1～2: 1, and spin coating rotating speed is 900～1200rpm, during spin coating
Between be 10s～12s.
Step 7, utilize prepared by fluorescence microscope detecting step six with gold-plated titanium dioxide/graphene oxide hydrogel one
Dimensional photonic crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
The silicon chip that following example use is found crystal silicon material company limited by Zhejiang and provides；SiO2Particle is biological by Southeast China University
Electronics National Key Laboratory provides.
Embodiment 1
1) silicon chip is immersed acetone soln sonic oscillation 30min, soaks sonic oscillation 30min in ethanol solution the most again,
It is finally immersed in the H that volume ratio is 1: 32SO4/H2O2Mixed solution soaks 12h, N2Dry up the sky obtaining cleaning up
White silicon chip.
2) weigh 4g butyl titanate to add in 16ml ethanol, be slowly added dropwise 2ml glacial acetic acid under magnetic agitation, under room temperature
Stirring 5h, obtains the pale yellow transparent TiO that concentration is 39mg/ml2Colloid solution.GO is soluble in water, it is made into concentration
For the GO colloid solution of 0.25mg/ml, the GO hydrogel of the 0.25mg/mL prepared is poured in autoclave,
90 DEG C of reaction 17h, obtain the GO hydrogel that concentration is 0.25mg/ml.
3) successively by TiO2Colloid solution and GO spin coating hydrogel are to substrate surface, and spin speed is 4500rpm, rotation
The painting time is 50s, and the spin coating cycle is 4 cycles, prepares titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
4) gold-plated on titanium dioxide/graphene oxide hydrogel 1-D photon crystal by the method for vacuum evaporation, evaporation
Time is 60s, obtains described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
5) the beta glucan solution taking 300 μ L is put in centrifuge tube, adds the 1mg/ml of 630 μ L in centrifuge tube
Aniline blue solution, 80 DEG C of water-bath 15min of lucifuge, obtain the glimmering of aniline blue formation specific binding with beta glucan
Photoreactivation product.
6) obtained fluorescence combination product is mixed by 1: 1 with 20mg/ml polyvinyl alcohol, utilize spin-coating method by above-mentioned
Solution is coated onto gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal surface, and spin coating rotating speed is 1000rpm,
Spin-coating time is 12s.
7) utilize prepared by fluorescence microscope detecting step six with gold-plated titanium dioxide/one-dimensional light of graphene oxide hydrogel
Sub-crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
As it is shown in figure 1, the gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal prepared for the present embodiment
Fluorescence Increasing effect.Can be the most weak with the fluorescence intensity of spin coating fluorescence combination product on the glass sheet from figure, for
29.67；The fluorescence intensity of the fluorescence combination product being spin-coated on titanium dioxide/graphene oxide hydrogel 1-D photon crystal
It is in centre, is about 8 times of fluorescence intensity of spin coating fluorescence combination product on the glass sheet, is 244.9；Spin coating
The fluorescence intensity of the fluorescence combination product on gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal is the strongest,
It is about 10 times of fluorescence intensity of spin coating fluorescence combination product on the glass sheet, is 314.38.
Fig. 2 is that gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal of preparing of embodiment 1 is for examining
Survey the result figure of beta glucan.It appeared that along with the rising of beta glucan concentration, fluorescence intensity is also progressively from figure
Strengthen, and there is certain linear relationship.
Embodiment 2
1) silicon chip is immersed acetone soln sonic oscillation 30min, immerses sonic oscillation 30min in ethanol solution the most again,
It is finally immersed in the H that volume ratio is 1: 32SO4/H2O2Mixed solution soaks 4h, N2Dry up the blank obtaining cleaning up
Silicon chip.
2) weigh 4g butyl titanate to add in 16ml ethanol, be slowly added dropwise 2ml glacial acetic acid under magnetic agitation, under room temperature
Stirring 6h, obtains the TiO that concentration is 39mg/ml2Colloid solution, then dilution 3 times, obtaining concentration is 13mg/ml's
TiO2Colloid solution.GO is soluble in water, it is made into the GO colloid solution that concentration is 0.5mg/ml, by prepare
The GO hydrogel of 0.5mg/mL is poured in autoclave, 90 DEG C of reaction 18h, obtains the GO water that concentration is 0.5mg/ml
Gel.
3) successively by TiO2Colloid solution and GO spin coating hydrogel are to substrate surface, and spin speed is 5000rpm, spin coating
Time is 50s, and the spin coating cycle is 4 cycles, prepares titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
4) gold-plated on titanium dioxide/graphene oxide hydrogel 1-D photon crystal by the method for vacuum evaporation, evaporation
Time is 90s, obtains described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
5) the beta glucan solution taking 300 μ L is put in centrifuge tube, adds the 1mg/ml benzene of 630 μ L in centrifuge tube
Amine indigo plant solution, 50 DEG C of water-bath 30min of lucifuge, obtain aniline blue multiple with the fluorescence of the specific binding formation of beta glucan
Close product.
6) obtained fluorescence combination product is mixed by 1: 1 with 40mg/ml polyvinyl alcohol, utilize spin-coating method by above-mentioned molten
Liquid is coated onto gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal surface, and spin coating rotating speed is 1200rpm, rotation
The painting time is 10s.
7) utilize prepared by fluorescence microscope detecting step six with gold-plated titanium dioxide/one-dimensional light of graphene oxide hydrogel
Sub-crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
Embodiment 3
1) silicon chip is immersed acetone soln sonic oscillation 60min, immerses sonic oscillation 60min in ethanol solution the most again,
It is finally immersed in the H that volume ratio is 3: 72SO4/H2O2Mixed solution soaks 12h, N2Dry up the sky obtaining cleaning up
White silicon chip.
2) weigh 4g butyl titanate to add in 16ml ethanol, be slowly added dropwise 2ml glacial acetic acid under magnetic agitation, under room temperature
Stirring 5h, obtains the TiO that concentration is 39mg/ml2Colloid solution, then dilution 5 times, obtaining concentration is 7.8mg/ml's
TiO2Colloid solution.GO is soluble in water, it is made into the GO colloid solution that concentration is 0.25mg/ml, by prepare
The GO hydrogel of 0.25mg/mL is poured in autoclave, and 100 DEG C of reaction 17h, obtaining concentration is 0.25mg/ml's
GO hydrogel.
3) successively by TiO2Colloid solution and GO spin coating hydrogel are to substrate surface, and spin speed is 6000rpm, rotation
The painting time is 60s, and the spin coating cycle is 4 cycles, prepares titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
4) gold-plated on titanium dioxide/graphene oxide hydrogel 1-D photon crystal by the method for vacuum evaporation, evaporation
Time is 60s, obtains described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal.
5) the beta glucan solution taking 300 μ L is put in centrifuge tube, adds the 1mg/ml of 630 μ L in centrifuge tube
Aniline blue solution, 80 DEG C of water-bath 20min of lucifuge, obtain the glimmering of aniline blue formation specific binding with beta glucan
Photoreactivation product.
6) obtained fluorescence combination product is mixed by 1: 1 with 20mg/ml polyvinyl alcohol, utilize spin-coating method by above-mentioned
Solution is coated onto gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal surface, and spin coating rotating speed is 900rpm,
Spin-coating time is 12s.
7) utilize prepared by fluorescence microscope detecting step six with gold-plated titanium dioxide/one-dimensional light of graphene oxide hydrogel
Sub-crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
Claims (6)
1. a beta glucan detection method based on Fluorescence Increasing principle, it is characterised in that this detection method include as
Lower step:
Step one, prepare titanium dioxide TiO respectively2Colloid solution and graphene oxide GO hydrogel, wherein, TiO2
Colloid solution concentration is 7.8～39mg/ml, and GO hydrogel concentration is 0.25～0.5mg/ml；
Step 2, silicon chip are cleaned and are dried up, and utilize the TiO that step one is prepared by spin-coating method2Colloid solution and GO hydrogel
It is coated onto silicon chip surface successively, obtains described titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, every layer of rotation
Being coated with rotating speed is 3000～6000rpm, and spin-coating time is 50s～1min；
Step 3, by the method for vacuum evaporation on titanium dioxide/graphene oxide hydrogel 1-D photon crystal gold-plated,
Obtaining described gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal, wherein, the evaporation time is 60s～90s；
Step 4, beta glucan solution is specific binding with aniline blue solution, obtain fluorescence combination product, wherein,
Beta glucan solution concentration is 1～6 μ g/ml, and aniline blue solution concentration is 0.5～1.5mg/ml；
Step 5, it is 1: 1～2: 1 to mix the fluorescence combination product obtained by step 4 and polyvinyl alcohol by volume ratio,
Utilize spin-coating method to be coated onto by above-mentioned solution gold-plated titanium dioxide/one-dimensional photon of graphene oxide hydrogel that step 3 obtains
Plane of crystal, obtains described with gold-plated titanium dioxide/graphene oxide hydrogel 1-D photon crystal for Fluorescence Increasing substrate
Fluorescence combination product, wherein, poly-vinyl alcohol solution concentration is 20～40mg/ml, and spin coating rotating speed is 900～1200rpm,
Spin-coating time is 10s～12s；
Step 6, utilize prepared by fluorescence microscope detecting step five with gold-plated titanium dioxide/graphene oxide hydrogel one
Dimensional photonic crystal is the fluorescence spectrum of the fluorescence combination product of Fluorescence Increasing substrate.
Beta glucan detection method based on Fluorescence Increasing principle the most according to claim 1, it is characterised in that
TiO described in step one2The preparation of colloid solution comprises the steps: that weighing butyl titanate adds in ethanol, and magnetic force stirs
Mixing down and be slowly added dropwise glacial acetic acid, wherein butyl titanate, the volume ratio of ethanol and glacial acetic acid is 2: 8: 1, stirs under room temperature
5～6h, obtain TiO2Colloid solution, dilutes 1～5 times, obtains the TiO that concentration is 7.8～39mg/ml2Colloid solution.
Beta glucan detection method based on Fluorescence Increasing principle the most according to claim 1, it is characterised in that
The preparation of GO hydrogel described in step one comprise the steps: by prepare 0.25～0.5mg/mL GO hydrogel
Pour in autoclave, 90～100 DEG C of reactions 17～24h, obtain the GO hydrogel that concentration is 0.25～0.5mg/ml.
Beta glucan detection method based on Fluorescence Increasing principle the most according to claim 1, it is characterised in that
Silicon chip described in step 2 is cleaned to dry up and is comprised the steps: silicon chip immerses acetone soln sonic oscillation 30～60min, it
After immerse sonic oscillation 30～60min in ethanol solution again；It is finally immersed in the H that volume ratio is 3: 7～1: 32SO4/H2O2Mixed
Close and solution soaks 4～12h, N2Dry up the silicon chip obtaining cleaning up.
Beta glucan detection method based on Fluorescence Increasing principle the most according to claim 1, it is characterised in that
To be coated with one layer of TiO in step 22Colloid solution and one layer of GO hydrogel are a spin coating cycle, and the described spin coating cycle is
3～4 cycles.
Beta glucan detection method based on Fluorescence Increasing principle the most according to claim 1, it is characterised in that
In step 4, with aniline blue is specific binding, beta glucan comprises the steps: that taking beta glucan solution puts in centrifuge tube,
Adding aniline blue solution in centrifuge tube, wherein the volume ratio of beta glucan solution and aniline blue solution is 1: 21, lucifuge
50～80 DEG C of water-baths 15～30min, obtain the fluorescence combination product of aniline blue and the specific binding formation of beta glucan.
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| CN109724961A (en) * | 2019-01-21 | 2019-05-07 | 陕西科技大学 | A kind of method of photonic crystal fluorescence enhancement detection trace organic amine compound |
| CN112691074A (en) * | 2021-03-24 | 2021-04-23 | 季华实验室 | Optical metamaterial, application and macromolecular substance trans-epidermal cell introduction method |
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| CN109724961A (en) * | 2019-01-21 | 2019-05-07 | 陕西科技大学 | A kind of method of photonic crystal fluorescence enhancement detection trace organic amine compound |
| CN112691074A (en) * | 2021-03-24 | 2021-04-23 | 季华实验室 | Optical metamaterial, application and macromolecular substance trans-epidermal cell introduction method |
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