CN114231268B - Non-contact cholesterol sensor for enhancing rare earth doped up-conversion nanoparticle luminescence based on photonic crystal effect and preparation method thereof - Google Patents
Non-contact cholesterol sensor for enhancing rare earth doped up-conversion nanoparticle luminescence based on photonic crystal effect and preparation method thereof Download PDFInfo
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- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 title claims abstract description 145
- 235000012000 cholesterol Nutrition 0.000 title claims abstract description 68
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 54
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- 230000000694 effects Effects 0.000 title claims abstract description 20
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- 229910052761 rare earth metal Inorganic materials 0.000 title abstract description 9
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- 239000013078 crystal Substances 0.000 claims abstract description 25
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- 238000000034 method Methods 0.000 claims description 9
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 3
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 3
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- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- VVQNEPGJFQJSBK-UHFFFAOYSA-N Methyl methacrylate Chemical compound COC(=O)C(C)=C VVQNEPGJFQJSBK-UHFFFAOYSA-N 0.000 description 2
- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
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- 108010089254 Cholesterol oxidase Proteins 0.000 description 1
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
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- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
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- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
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- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/77—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals
- C09K11/7766—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals containing two or more rare earth metals
- C09K11/7772—Halogenides
- C09K11/7773—Halogenides with alkali or alkaline earth metal
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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"
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Abstract
Polymethyl methacrylate (PMMA) protein Dan Guangzi crystal effect enhanced LiErF 4 :0.5％Tm 3+ @LiYF 4 A non-contact cholesterol sensor with rare earth doped up-conversion nano particle luminescence and a preparation method thereof belong to the technical field of fluorescence sensors. The invention utilizes the rotary spraying method to lead LiErF 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particles are compounded with PMMA photonic crystals, and the coupling effect of the photonic crystal band gap and the excitation light field is utilized to enhance the excitation light field so as to lead LiErF 4 :0.5％Tm 3+ @LiYF 4 The luminescence intensity of the up-conversion nano particles is obviously improved, so that the sensitivity of cholesterol detection is improved, and a non-contact fluorescence sensor is constructed. The fluorescence sensor has the advantages of high sensitivity, good specificity and strong stability for cholesterol detection, and has wide application prospect in whole blood detection.
Description
Technical Field
The invention belongs to the technical field of fluorescence sensors, and in particular relates to a polymethyl methacrylate (PMMA) protein Dan Guangzi crystal effect enhanced LiErF 4 :0.5％Tm 3+ @LiYF 4 The non-contact cholesterol sensor with rare earth doped up-conversion nano particle luminescence and the preparation method thereof can be applied to the aspect of detecting cholesterol in serum.
Background
Cholesterol is an indispensable essential substance for human tissue cells and plays a vital role in basic cell life activities. The serum cholesterol level of the healthy person is 2.60-5.20 mmol/L, and the research result shows that the high level of cholesterol in the blood of the human body can cause atherosclerosis and cardiovascular and cerebrovascular diseases; while low levels of cholesterol increase the risk of cerebral hemorrhage and reduce body resistance. Cholesterol levels are important indicators for clinical biochemical examinations and prevention of cardiovascular disease. Therefore, measuring cholesterol in blood is of great practical importance for maintaining physical health.
Currently cholesterol detection is colorimetric, high performance liquid chromatography, electrochemical analysis, chemiluminescence and fluorescence. Among them, fluorescence technology has become an important method for real-time detection of cholesterol in living cells, serum and tissues. Although many developments have been made in this method, most of the excitation light sources of the organic dyes and nano luminescent materials used are ultraviolet or visible light, and the biomolecules have strong absorption in this area, so that the biological autofluorescence is easy to generate, and the detection sensitivity is seriously affected. Thus, it remains a challenge to explore a highly sensitive, interference-free and recyclable fluorescence method to monitor cholesterol levels.
The rare earth doped up-conversion fluorescent material nano-particles have received a great deal of attention in the construction of fluorescent sensors because of the advantages of no autofluorescence, no bleaching phenomenon and deep tissue penetration. However, due to the small absorption cross section of rare earth ions, the low fluorescence quantum yield limits their further application in bioluminescence sensors. It is therefore important to increase the up-conversion fluorescence intensity to increase the sensitivity of the biological detection. The use of photonic crystal effects has become an effective means of increasing the up-conversion luminescence intensity. Therefore, a fluorescent probe or system based on the photonic crystal effect enhanced rare earth doped fluorescent material is developed, is used for high-sensitivity detection of cholesterol in serum, and has great practical significance and application prospect.
Disclosure of Invention
The invention aims to provide a method for enhancing LiErF based on PMMA protein Dan Guangzi crystal effect 4 :0.5％Tm 3+ @LiYF 4 A non-contact cholesterol sensor with rare earth doped up-conversion nano particle luminescence and a preparation method thereof. The present invention aims to enhance by utilizing the photonic crystal effectUp-conversion luminescence, and further detection sensitivity is improved. By constructing a non-contact sensor, a method for detecting cholesterol fluorescence without interference and in recycling is provided, and the sensor can be applied to the aspect of detecting cholesterol in serum.
The invention firstly prepares LiErF 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particle and PMMA protein Dan Guangzi crystal composite film enhances up-conversion luminescence so as to improve detection sensitivity, and a non-contact fluorescence sensor is constructed. Based on 3,3', 5' -Tetramethylbenzidine (TMB) oxidation product (Ox TMB) to LiErF produced by cholesterol cascade reaction 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particles have strong quenching effect of near monochromatic red light, and cholesterol is quantitatively detected according to the linear relation between the change of red light intensity and cholesterol concentration. The fluorescence sensor has the advantages of high sensitivity, good specificity and strong stability for cholesterol detection, and has wide application prospect in whole blood detection.
The invention relates to a PMMA protein Dan Guangzi crystal effect-based LiErF enhanced method 4 :0.5％Tm 3+ @LiYF 4 The preparation method of the non-contact cholesterol sensor with up-conversion nano particle luminescence comprises the following steps:
(1) LiErF with core-shell structure 4 :0.5％Tm 3+ @LiYF 4 Synthesis of up-conversion nanoparticles: liErF with particle size of 20-25 nm is prepared by a high-temperature solvothermal method 4 :0.5％Tm 3+ Bare core, then in LiErF 4 :0.5％Tm 3+ Epitaxial growth of LiYF on bare nuclei 4 Inert shell layer to obtain LiErF with particle diameter of 30-35 nm 4 :0.5％Tm 3+ @LiYF 4 Up-converting nanometer particle with core-shell structure and final LiErF reaction 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nanoparticles are dispersed into a cyclohexane solution;
(2) Preparation of PMMA protein Dan Guangzi crystal film: preparing a monodisperse PMMA microsphere solution with the average diameter of the microspheres between 350 and 450nm, vertically inserting a clean glass slide into the monodisperse PMMA microsphere solution, keeping the temperature of 30 to 40 ℃ for 20 to 25 hours, taking out the glass slide, drying the glass slide, and heating the dried glass slide for 40 to 60 minutes at the temperature of 120 to 140 ℃ to obtain a PMMA protein Dan Guangzi crystal film on the glass slide;
(3) PMMA protein Dan Guangzi crystal and LiErF 4 :0.5％Tm 3+ @LiYF 4 Compounding of up-conversion nanoparticles: fixing PMMA protein Dan Guangzi crystal film on a rotary table, and under the rotation speed of 400-600 r/s, liErF with the concentration of 0.5-1.5 mL and 0.5-1.0 mmol/mL is prepared 4 :0.5％Tm 3+ @LiYF 4 Spraying a cyclohexane solution of the up-conversion nano particles onto the PMMA protein Dan Guangzi crystal film; liErF as cyclohexane evaporates 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particles are self-assembled on the photonic crystal film, and a composite film of the photonic crystal and the up-conversion nano particles is obtained on a glass slide;
(4) Covering an ultrathin glass plate with the thickness of 50-150 mu m on the composite film of the photonic crystal and the up-conversion nano particles to isolate the liquid to be detected from the composite film, and adhering an acrylic plate with square holes on the ultrathin glass plate to contain the liquid to be detected, wherein the size of the acrylic plate is smaller than or equal to that of the ultrathin glass plate, thereby obtaining the PMMA protein Dan Guangzi crystal effect-based enhanced LiErF 4 :0.5％Tm 3+ @LiYF 4 A non-contact cholesterol sensor for up-converting nanoparticle luminescence.
The invention relates to a PMMA protein Dan Guangzi crystal effect-based LiErF enhanced method 4 :0.5％Tm 3+ @LiYF 4 The non-contact cholesterol sensor with the up-conversion nano particle luminescence is prepared by the method.
Working principle:
the invention utilizes the photonic crystal effect to strengthen LiErF 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particles emit light, so that the detection sensitivity is improved. Under 980nm excitation, oxidation products (Ox TMB) based on 3,3', 5' -Tetramethylbenzidine (TMB) produced by cholesterol cascade quench LiErF 4 :0.5％Tm 3+ @LiYF 4 Strong near monochromatic red light, linearly dependent on cholesterol concentration according to the change of fluorescence intensityThe quantitative detection of cholesterol in the solution and serum is realized.
The LiErF prepared by the invention is enhanced based on photonic crystal effect 4 :0.5％Tm 3+ @LiYF 4 The non-contact cholesterol fluorescence sensor for up-conversion nanoparticle luminescence has the following advantages:
1. LiErF using photonic crystal effect 4 :0.5％Tm 3+ @LiYF 4 The luminous intensity of the up-conversion nano particles is enhanced by tens of times, and the sensitivity of cholesterol detection is improved.
2. Due to LiErF 4 :0.5％Tm 3+ @LiYF 4 Middle Er 3+ The ions have rich step-shaped energy levels, can generate near-monochromatic red light emission under the excitation of various lasers (800 nm, 980nm and 1530 nm), can widen an excitation light source, and is beneficial to the wide application of cholesterol detection.
3. The test adopts excitation light (980 nm) in near infrared band, can eliminate background interference from biological sample, improve detection sensitivity, and has good selectivity and stability.
4. The non-contact sensor can realize non-contact nondestructive detection of serum samples without interference and pollution, can be recycled, simplifies the detection method and greatly reduces the cost.
Drawings
Fig. 1: the invention relates to a structural schematic diagram of a non-contact cholesterol sensor based on photon crystal effect enhanced up-conversion nano particles; the names of the components are as follows: the device comprises a glass slide 1, a photonic crystal film 2, up-conversion nano particles 3, an ultrathin glass plate 4, an acrylic plate 5 with square holes and square holes 6.
Fig. 2: liErF on non-contact cholesterol sensor in detection example 1 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nanoparticle up-converts luminescence with cholesterol concentration profile under 980 excitation.
Fig. 3: liErF on non-contact cholesterol sensor in detection example 1 4 :0.5％Tm 3+ @LiYF 4 Up-conversion nanoparticles were quenched by red light intensity at 980 excitation (F/F) 0 ) A linear graph of cholesterol concentration, wherein F/F 0 The ratio of the integral intensity of the up-conversion red light emission after adding cholesterol and without adding cholesterol into the system is obtained.
Fig. 4: statistical graphs of interference detection results of detection example 3.
Fig. 5: the schematic diagram of the present invention. Drop-adding the test solution containing different concentrations of cholesterol prepared in test example 1 onto the non-contact cholesterol sensor prepared in example 3, under 980nm excitation, liErF 4 :0.5％Tm 3+ @LiYF 4 The red light of the up-conversion nanoparticle is quenched by Ox TMB generated by the cholesterol cascade reaction, and the higher the cholesterol concentration, the more Ox TMB generated, and the more pronounced the quenching. According to the linear relation between the change of fluorescence intensity and cholesterol concentration, the quantitative detection of cholesterol in the solution and serum is realized.
Detailed Description
Example 1: liErF 4 :0.5％Tm 3+ @LiYF 4 Synthesis of upconverting nanoparticles
In the following examples, liErF 4 :0.5％Tm 3+ @LiYF 4 The upconverting nanoparticles were prepared by a high temperature solvothermal method as described in the reference (Li, z., zhang, y., nanotechnology,2008.19,345606).
(1) TmCl is firstly taken up 3 ·6H 2 O(0.005mmol)、ErCl 3 ·6H 2 O (0.995 mmol), 14mL Octadecene (ODE) and 6mL Oleic Acid (OA) were mixed in a 50mL flask and stirred for 30 min. The solution was warmed to 160℃for 40 minutes and then cooled to 50 ℃. 10mL of a mixture containing LiOH (2.500 mmol) and NH was added 4 F (4.000 mmol) in methanol and heated to 70℃for 40 min. The mixture was then warmed to 300℃for 90 minutes and cooled to 50℃and centrifuged with acetone and ethanol to give LiErF 4 :0.5％Tm 3+ And (5) bare core. Finally, liErF is carried out 4 :0.5％Tm 3+ The bare nuclei were dissolved in 8mL of cyclohexane for use. Note that continuous nitrogen flow and agitation was necessary throughout the experiment.
(2) YCl is combined with 3 ·6H 2 A mixture of O (0.750 mmol), 10mL ODE and 5mL OA at 50mix and stir in the mL flask for 30 minutes. The solution was warmed to 160 ℃ for 40 minutes and then cooled to 50 ℃. Then 4mL of the solution containing LiErF prepared in the step (1) was added 4 :0.5％Tm 3+ The bare core cyclohexane solution was reacted for 30 minutes at 85℃and then cooled to 50 ℃.5mL of a mixture containing LiOH (1.875 mmol) and NH was added 4 F (3.000 mmol) in methanol and heated to 70℃for 30 min. Heating the solution to 300 ℃ for 60 minutes, cooling to 50 ℃, washing with acetone and ethanol, and centrifuging to obtain LiErF 4 :0.5％Tm 3+ @LiYF 4 Up-converting the nano particles in the core-shell structure; finally LiErF is added 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nanoparticles were dissolved in 4mL of cyclohexane for use.
Example 2: preparation of PMMA photonic crystal structure
In the following examples, photonic crystals were synthesized by the vertical deposition method described in references (Yin, z, li, h, xu, w, cui, s, methou, d, chen, x, zhu, y, qin, g, song, h, 2016.Advanced materials 28,2518 to 2525).
With 100mL, 5mg mL -1 20mL of Methyl Methacrylate (MMA) was washed 7 times with aqueous NaOH, then 6mL of the washed MMA was mixed with 80mL of deionized water and heated, and 36mg of K was added 2 S 2 O 8 The reaction was carried out at 90℃for 90 minutes to give a monodisperse PMMA microsphere solution having an average diameter size of 424 nm. The washed slide glass was vertically inserted into a PMMA microsphere solution, kept at 35 ℃ for 24 hours, dried, and then heated to 130 ℃ for 60 minutes, thereby obtaining a PMMA protein Dan Guangzi crystal film on the slide glass.
Example 3: preparation of photon crystal effect enhanced rare earth doped up-conversion luminescence non-contact cholesterol sensor
The PMMA protein Dan Guangzi crystal film prepared in example 2 was fixed together with a glass slide on a rotary table, the rotary table was started at 500r/s, and 1mL and 0.8mmol mL were sprayed with a spray gun -1 Example 1 LiErF prepared 4 :0.5％Tm 3+ @LiYF 4 Spraying cyclohexane solution of up-conversion nano particles onto photonic crystal film. LiErF as cyclohexane evaporates 4 :0.5％Tm 3+ @LiYF 4 The up-conversion nano particles are self-assembled on the surface of the photonic crystal film to obtain the PMMA protein Dan Guangzi crystal and up-conversion nano particle composite film. And then covering an ultrathin glass plate with the thickness of 100 mu m, the length of 7.5cm and the width of 2.5cm on the composite film of the photonic crystal and the up-conversion nano particles to isolate the liquid to be detected from the composite film, and pasting an acrylic plate with the length of 6.0cm, the width of 2.5cm and the thickness of 0.3cm and with square holes with the side length of 0.6cm on the ultrathin glass plate to contain the liquid to be detected, thereby obtaining the non-contact cholesterol sensor for enhancing up-conversion based on the photonic crystal effect.
Detection example 1: detection of cholesterol content by non-contact sensor
mu.L of phosphate buffer (0.1M, pH 7.0), 100. Mu.L of cholesterol oxidase (0.16U mL) -1 ) 200. Mu.L of deionized water and 100. Mu.L of cholesterol solutions (dissolved in methanol) of different concentrations (0. Mu.M, 2. Mu.M, 4. Mu.M, 6. Mu.M, 8. Mu.M, 10. Mu.M, 20. Mu.M, 30. Mu.M, 40. Mu.M, 80. Mu.M, 100. Mu.M, 200. Mu.M) were mixed and incubated at 37℃for 30 minutes, and 100. Mu.L of acetic acid buffer (0.1M, pH 4.0), 100. Mu.L of horseradish peroxidase (6 mU mL) were added -1 ) 100. Mu.L of 20mM TMB and 1.2mL deionized water were added to the above mixture at 37℃for 30 minutes. The resulting mixed solution was injected into a square hole as described in example 3, and LiErF was monitored using a fluorescence spectrometer model Quant Master 8000 4 :0.5％Tm 3+ @LiYF 4 Up-conversion nanoparticles up-convert emission spectra on a non-contact cholesterol sensor. The results are shown in FIG. 2, where the red light intensity gradually decreases with increasing cholesterol concentration. Simultaneous cholesterol concentration and red light intensity quenching (F/F) 0 ) Has better linear relation between 2 and 30 mu M and meets the F/F 0 Equation (wherein F/F) of =0.942-0.023C (μm) 0 The ratio of the integrated intensities of the red emissions for the up-conversion of cholesterol and no cholesterol in the system, C is the cholesterol concentration in μm), as shown in fig. 3.
Detection example 2: detection of cholesterol in serum
Human serum for use in the present invention is provided by the first hospital friend of Jilin university.
Table 1: detection of cholesterol levels in human serum samples using a non-contact cholesterol sensor
Human serum was diluted 50-fold with deionized water, and serum samples 1, 2 and 3 after dilution were used in place of the cholesterol solution in test example 1, and cholesterol in serum was detected by the method of test example 1. The results are shown in table 1, and the relative standard deviation (rsd=standard deviation/average) between the cholesterol detection concentration measured by the non-contact cholesterol fluorescence sensor of the present invention and the cholesterol concentration (clinical data) measured by the enzyme-linked immunosorbent assay used clinically is lower than 11.0%, which indicates that the cholesterol sensor of the present invention can be used for quantitatively detecting cholesterol in complex serum samples.
Detection example 3: specific detection of cholesterol content by non-contact sensor
Similar to test example 1, test specificity was verified by adding 2mM glucose, urea, uric acid, ascorbic acid, cysteine, magnesium chloride and 20. Mu.M and 100. Mu.M cholesterol, respectively, instead of cholesterol in test example 1, and the test results are shown in FIG. 3. From the graph, only the cholesterol-added sample realizes the reduction of the red light up-conversion emission spectrum, and other samples have no influence on the fluorescence spectrum signal, so that the specific detection of the cholesterol can be realized by the experimental scheme.
Claims (2)
1. LiErF is strengthened based on polymethyl methacrylate protein Dan Guangzi crystal effect 4 :0.5%Tm 3+ @LiYF 4 The preparation method of the non-contact cholesterol sensor with up-conversion nano particle luminescence comprises the following steps:
(1) LiErF with core-shell structure 4 :0.5%Tm 3+ @LiYF 4 Synthesis of up-conversion nanoparticles: first prepared by high-temperature solvothermal methodLiErF with particle size of 20-25 nm 4 :0.5%Tm 3+ Bare core, then in LiErF 4 :0.5%Tm 3+ Epitaxial growth of LiYF on bare nuclei 4 An inert shell layer is adopted to obtain LiErF with the particle size of 30-35 nm 4 :0.5%Tm 3+ @LiYF 4 Up-converting nanometer particle with core-shell structure and final LiErF reaction 4 :0.5%Tm 3+ @LiYF 4 The up-conversion nanoparticles are dispersed into a cyclohexane solution;
(2) Preparation of polymethyl methacrylate protein Dan Guangzi crystal film: preparing a monodisperse polymethyl methacrylate microsphere solution with the average diameter of the microsphere between 350 and 450nm, vertically inserting a clean glass slide into the monodisperse polymethyl methacrylate microsphere solution, keeping the temperature of 30 to 40 ℃ for 20 to 25 hours, taking out the glass slide, drying the glass slide, and heating the dried glass slide at 120 to 140 ℃ for 40 to 60 minutes to obtain a polymethyl methacrylate protein Dan Guangzi crystal film on the glass slide;
(3) Polymethyl methacrylate protein Dan Guangzi crystal and LiErF 4 :0.5%Tm 3+ @LiYF 4 Compounding of up-conversion nanoparticles: fixing a polymethyl methacrylate protein Dan Guangzi crystal film on a rotary table, and at the rotating speed of 400-600 r/s, preparing LiErF with the concentration of 0.5-1.5 mL and 0.5-1.0 mmol/mL 4 :0.5%Tm 3+ @LiYF 4 Spraying the cyclohexane solution of the up-conversion nano particles on a polymethyl methacrylate protein Dan Guangzi crystal film; liErF as cyclohexane evaporates 4 :0.5%Tm 3+ @LiYF 4 The up-conversion nano particles are self-assembled on the photonic crystal film, and a composite film of the photonic crystal and the up-conversion nano particles is obtained on a glass slide;
(4) An ultrathin glass plate with the thickness of 50-150 mm is covered on a composite film of the photonic crystal and the up-conversion nano particles so as to isolate liquid to be detected from the composite film, an acrylic plate with square holes is adhered on the ultrathin glass plate so as to contain the liquid to be detected, and the size of the acrylic plate is smaller than or equal to that of the ultrathin glass plate, so that the crystal effect enhanced LiErF based on polymethyl methacrylate protein Dan Guangzi is obtained 4 :0.5%Tm 3+ @LiYF 4 Up-conversionA non-contact cholesterol sensor with nanoparticle luminescence.
2. LiErF is strengthened based on polymethyl methacrylate protein Dan Guangzi crystal effect 4 :0.5%Tm 3+ @LiYF 4 The non-contact cholesterol sensor with up-conversion nanoparticle luminescence is characterized in that: is prepared by the method of claim 1.
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