CN101216415A - Photon crystal composite encoding microsphere and preparation method - Google Patents
Photon crystal composite encoding microsphere and preparation method Download PDFInfo
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Abstract
A photonic crystal compound-coding microsphere and preparation method relates to a method for compound-coding microsphere from a specific light reflection peak of photonic crystal as well as characteristic emission peak of quantum dots adsorbed by an outer layer and the intensity of the characteristic emission peak. The method is characterized in that the microsphere is made of colloid particles; quantum dots wrap the surface of the photonic crystal microsphere layer by layer; and quantum dot coding can be integrated with photonic crystal microsphere coding by wraping different quantum dots and different layers of quantum dots, thus improving the coding quantity. The polymer microsphere has the advantages of good coding stability, easy control, low production cost, simple operation, high efficiency, large detection flux, etc.
Description
Technical field
The present invention relates to a kind of microballoon by photonic crystal special light reflection peak and the quantum dot feature emission peak of outer absorption and the method that feature emission peak intensity is come composite coding, belong to the technical field of biomedical research, environmental monitoring and clinical detection.
Background technology
After human genomic sequence was decoded, the interaction between the function of gene and the protein of gene code became the focus of research, and the mankind have entered the genome times afterwards comprehensively.The distinguishing feature of genome times afterwards comprehensively is exactly will study between a large amount of albumen and nucleic acid, between the protein-protein and the interaction between protein and the medicine, these Study of Interaction almost are impossible adopting traditional test method, therefore need a kind of high-throughout test platform.High-throughout test platform has then required high-throughout molecular vehicle, identifies the difference interaction that different molecules takes place, promptly so-called cataloged procedure.Array biochip utilize diverse location that molecule fixes just coordinate distinguish different biomolecule as coding, but because the coordinate of array biochip can only be to be fixed in the two-dimensional space, therefore limited the speed and the application thereof of reaction.Novel in recent years code carrier occurs in succession.The making of polymer microballoon and application have had the history of decades, and its particle size expands to Nano grade from micron level always.But as a kind of solid phase code carrier and mark, microballoon becomes the focus that fields such as high flux screening and combinatorial chemistry are paid close attention to.
With respect to other forms of solid phase carrier, microballoon has significant advantage: the first, and the specific surface area of microballoon is big, can increase the effecting reaction surface to volume ratio, and the chemical reaction on surface is carried out in littler volume; The second, adopt microballoon can utilize some other supplementary meanss to realize a kind of reaction system between solid-liquid reaction and liquid-liquid reactions, thereby accelerate the reaction velocity of system as stir, liquid washes away etc. as carrier; The 3rd, the molecule of microsphere surface combination can be separated from solution after reaction is finished easily; The 4th, along with the change of microsphere surface functionalization group, can expand the purposes of microballoon.
The code carrier of microballoon mainly contains fluorescence-encoded micro-beads, quantum point coding microball, nano strip font code coding microball, combinatorial chemistry coding microball, photonic crystal coding particle, colloid bar coding etc. at present.Fluorescence-encoded micro-beads is that two kinds of different fluorescent dyes are mixed in the micro polymer bulb matrix according to accurate ratio, according to the difference of the ratio of these two kinds of fluorescent dyes in the microballoon microballoon is divided into not of the same race, thereby realize the coding of microballoon.The nano strip font code can be any molecule or the compound that the enough SPM of energy differentiate, such as carbon nano-tube, fullerene, sub-micron metal bar code, nano particle or quantum dot.The preparation relative complex of these coded systems and volume carrier, and the checkout equipment complexity of decode procedure needs, cost is higher.In addition, employing fluorochrome coding also exists photofading, radiative emission peak is difficult to problems such as differentiation and biocompatibility.Quantum point coding microball then is to utilize the different fluorescence of semiconductor nanocrystal luminescent quantum dot to encode with different fluorescence intensities.Compare with traditional organic fluorescence molecule, quantum dot has obvious superiority: exciting light spectrum width, emission spectrum be symmetrically distributed and width narrow, color tunable, the photochemical stability height is difficult for photodissociation.Quantum dot has continuous excitation spectrum, and size that can be by regulating crystal grain is to obtain continuous emission spectrum.The emission spectrum of quantum dot is very narrow, so only just can excite the quantum dot of multiple size simultaneously with a kind of light of wavelength, so, just can obtain the emission light of multiple color with minimum spectrum band gap.
The present invention combines quantum-dot coding and photonic crystal coding exactly, has increased encoding amount greatly, if the quantum dot of m kind feature emission peak has n kind feature emission peak intensity, so just can obtain n
m-a kind of coding, can there be the photon crystal micro-ball of k kind different coding the while, and encoding amount just can reach k (n so
m-1) plants.And quantum dot being fixed on the microballoon outside surface with the method for absorption layer by layer, the preparation method is simple, is easy to detect, and simplifies and detects step, reduces plurality of advantages such as detecting cost.
Summary of the invention
Technical matters: the preparation method who the purpose of this invention is to provide a kind of photon crystal composite encoding microsphere, this photon crystal micro-ball particle size range at 200 μ m between the 6mm, can be used as the carrier of high flux screening and multiple bioanalysis and detection, its coded system and preparation method are simply efficient, encoding amount is big, can reduce the complexity of checkout equipment and detect cost.Information such as concentration by can measuring testing molecule simultaneously to the detection of photon crystal composite encoding microsphere and kind.
Technical scheme: photon crystal composite encoding microsphere of the present invention can be realized by following scheme:
With the method for layer assembly quantum dot is wrapped in the photon crystal micro-ball surface of coding, by wrapping up the different quantum dots and the different quantum dot numbers of plies, makes quantum-dot coding combine with the photon crystal micro-ball coding, encoding amount increases greatly.Described photon crystal micro-ball is the opal structural photonic crystal of the orderly self assembly of colloidal particle, or the photonic crystal of counter opal structure.Described quantum dot is cadmium sulfide CdS, cadmium selenide CdSe, cadmium telluride CdTe or nucleocapsid quantum dot.Described nucleocapsid quantum dot is a zinc sulphide bag cadmium selenide CdSe@ZnS quantum dot.The photon crystal micro-ball of described coding is a kind of in polystyrene, polymethylmethacrylate, polyacrylonitrile, tygon, polypropylene, the polyethyl acrylate; Or be silicon dioxide microsphere or titanium dioxide microballoon sphere.
The preparation method of a kind of photon crystal composite encoding microsphere of the present invention specifically may further comprise the steps:
1) chooses the photon crystal micro-ball of different coding, it is carried out surface treatment, obtain electronegative photonic crystal coding microball; Wherein photon crystal micro-ball is the polyvinyls microballoon or is inorganic photon crystal micro-ball;
2) utilize the method for absorption layer by layer, earlier at the positive polyelectrolyte of photon crystal micro-ball surface adsorption, the negative polyelectrolyte of absorption then, adsorb positive polyelectrolyte again after, the quantum dot of different characteristic emission peak is adsorbed on the microballoon outside surface;
3) to adsorb the quantum dot of the different numbers of plies or variety classes size, layer with layer between separate with three strata electrolyte;
4) continue to adsorb four strata electrolyte again at outermost quantum dot, so that detect.
Described positive polyelectrolyte can be polypropylene amine hydrochloride PAH, also can be polyethyleneimine PEI; Used negative polyelectrolyte is kayexalate PSS, also can be polyacrylic acid PAA.Described quantum-dot coding is the feature emission peak of quantum dot, and the feature emission peak that utilizes different quantum dots is as different quantum-dot codings.Described quantum-dot coding is the intensity of the feature emission peak of quantum dot, controls the intensity of its feature emission peak by the quantum dot that adsorbs the different numbers of plies, and the intensity of utilizing the different characteristic emission peak is as different quantum-dot codings.Described quantum dot be encoded to its feature emission peak and its feature emission peak combination of strength, utilize the absorption variety classes and the different numbers of plies to control the intensity of feature emission peak He its feature emission peak of quantum dot.
Beneficial effect: utilize the method for the mixed multiple encoding of quantum-dot coding and photonic crystal according to the present invention, prepared photon crystal composite encoding microsphere, have the following advantages:
1) adopts the mixed multiple encoding of quantum-dot coding and photonic crystal, expanded encoding amount greatly,, so just can obtain n if the quantum dot of m kind feature emission peak has n kind feature emission peak intensity
m-a kind of coding can have the different photon crystal micro-ball of k kind simultaneously, and encoding amount just can reach k (n so
m-1) plants.Can satisfy the needs that detect a plurality of indexs simultaneously, also can satisfy the needs of high throughput testing;
2) coding/decoding method of the photon crystal micro-ball of quantum dot coating is simple, only just can excite the quantum dot of multiple size simultaneously with a kind of light of wavelength, come mark thing to be detected with quantum dots similar but different sizes simultaneously, the concentration that in decoding, also can read thing to be detected, therefore reaction finishes only need detect the feature reflection peak of quantum dot feature emission peak and photonic crystal, just can learn the kind and the content of testing molecule simultaneously, testing process is simple, and fast.
3) preparation method of this photon crystal composite encoding microsphere is simple, chooses the photon crystal micro-ball of different coding respectively, goes up the different numbers of plies and different types of quantum dot with absorption layer by layer again, control easily, and cost of manufacture is cheap, and is simple efficient;
4) the code used good stability of this photon crystal composite encoding microsphere, quantum dot exciting light spectrum width, emission spectrum be symmetrically distributed and width narrow, color tunable, the photochemical stability height is difficult for photodissociation.Therefore can there be phenomenons such as the photobleaching of fluorescent pigment and optical quenching, in application process, remain stable coding;
5) applied range can be after being prepared into photon crystal composite encoding microsphere, in its skin adsorbing electrolyte again, just can fix different function group, realizes the functionalization of microballoon, thereby the condition with the molecular chemistry coupling that will fix is provided.
Description of drawings
The synoptic diagram that Fig. 1 generates for the opal structural photon crystal micro-ball.
Fig. 2 utilizes the layer assembly legal system to be equipped with the synoptic diagram of photon crystal composite encoding microsphere.
Embodiment
Prepare a kind of photon crystal composite encoding microsphere, choose the photon crystal micro-ball of different coding, method with layer assembly is wrapped in the photon crystal micro-ball surface with quantum dot, by wrapping up the different quantum dots and the different quantum dot numbers of plies, make quantum-dot coding combine with the photon crystal micro-ball coding, encoding amount increases greatly.
The numerical value that is encoded to feature reflection peak wavelength in the photonic crystal reflectance spectrum of photon crystal micro-ball.Photon crystal micro-ball can be the opal structural photonic crystal of the orderly self assembly of colloidal particle, also can be the photonic crystal of counter opal structure.The coding of quantum dot can be its feature emission peak, also can be the intensity of its feature emission peak, can also be the intensity combination of its feature emission peak and its feature emission peak, control the intensity of feature emission peak He its feature emission peak of quantum dot by the absorption variety classes and the different numbers of plies.Quantum dot is generally water-soluble quantum dot, can be CdS, CdSe, CdTe and some nucleocapsid quantum dots such as CdSe@ZnS etc.Used positive polyelectrolyte can be polypropylene amine hydrochloride (PAH), also can be polyethyleneimine (PEI); Used negative polyelectrolyte is kayexalate (PSS), also can be polyacrylic acid (PAA).
The preparation method may further comprise the steps:
Choose the photon crystal micro-ball of different coding, it is carried out surface treatment, obtain electronegative photonic crystal coding microball;
Utilize the method for absorption layer by layer, earlier at the positive polyelectrolyte of photon crystal micro-ball surface adsorption, the negative polyelectrolyte of absorption then, adsorb positive polyelectrolyte again after, the quantum dot of different characteristic emission peak is adsorbed on the microballoon outside surface;
The quantum dot that adsorbs the different numbers of plies or variety classes size separates with three strata electrolyte between layer and the layer;
Continue to adsorb four strata electrolyte again at outermost quantum dot, so that detect.
Come mark thing to be detected with quantum dots similar but different sizes simultaneously, the concentration that in decoding, also can read thing to be detected, therefore reaction finishes only to need to detect quantum dot emission spectrum and photon crystal for transmitting peak, just can learn the kind and the content of testing molecule simultaneously.
Embodiment one.The preparation of water-soluble CdTe mark polystyrene microsphere
1 chooses the polystyrene microsphere of 600nm and 540nm coding respectively, because synthetic polystyrene particle has carboxyl, then Zhi Bei microsphere surface also has negative charge.
2 at first are immersed in the microballoon for preparing in the 0.5mol/L NaCl solution of 1mg/mL PAH, and standing adsorption 20 minutes with ultrapure water flushing three times, is washed unnecessary PAH off.Again microballoon is immersed in the 0.5mol/L NaCl solution of 1mg/mLPSS same standing adsorption centrifuge washing.After adsorbing one deck PAH again, microballoon is immersed in the CdTe that mercaptoacetic acid is modified
540In (emission spectrum is at 540nm) aqueous solution, standing adsorption is centrifuge washing after 60 minutes.
3 repeat above-mentioned steps.After adsorbing three strata electrolyte, adsorb second layer CdTe again
540By that analogy, adsorbing three layers of CdTe
540After, continue three layers of CdTe of parcel again
580Adsorb four strata electrolyte again at outermost quantum dot.
4 will wrap the coding microball mixing of being crossed, and obtain two kinds of not homospecific composite coding microballoons.The coding that is photon crystal micro-ball is respectively 600nm and 540nm, and quantum-dot coding is CdTe
540: CdTe
560=3: 3 photon crystal composite encoding microsphere.
Embodiment two.The preparation of water-soluble CdTe mark silicon dioxide microsphere
1 chooses 400nm respectively, and the silicon dioxide microsphere of 460nm and 540nm coding spends the night in the hydrogen peroxide solution with its immersion 70% (V/V0) sulfuric acid and 30%, makes the microsphere surface of preparation have negative charge.
2 at first are immersed in the microballoon for preparing in the 0.5mol/L NaCl solution of 1mg/mL PAH, and standing adsorption 20 minutes with ultrapure water flushing three times, is washed unnecessary PAH off.Again microballoon is immersed in the 0.5mol/L NaCl solution of 1mg/mLPSS same standing adsorption centrifuge washing.After adsorbing one deck PAH again, microballoon is immersed in the CdTe that mercaptoacetic acid is modified
540In (emission spectrum is at 540nm) aqueous solution, standing adsorption is centrifuge washing after 60 minutes.
3 repeat above-mentioned steps.After adsorbing three strata electrolyte, adsorb second layer CdTe again
540By that analogy, adsorbing three layers of CdTe
540After, continue five layers of CdTe of parcel again
580, and then adsorb three layers of CdTe
620Adsorb four strata electrolyte again at outermost quantum dot.
4 will wrap the coding microball mixing of being crossed, and obtain three kinds of not homospecific composite coding microballoons.The coding that is photon crystal micro-ball is respectively 400nm, and 460nm and 540nm, quantum-dot coding are CdTe
540: CdTe
580: CdTe
620=3: 5: 3 photon crystal composite encoding microsphere.
Embodiment three.The preparation of water-soluble CdTe mark silicon dioxide microsphere
1 chooses the silicon dioxide microsphere of 540nm coding, and it is immersed 70% (V/V
0) spend the night in sulfuric acid and 30% the hydrogen peroxide solution, make the microsphere surface of preparation have negative charge.
2 at first are immersed in the microballoon for preparing in the 0.5mol/L NaCl solution of 1mg/mL PAH, and standing adsorption 20 minutes with ultrapure water flushing three times, is washed unnecessary PAH off.Again microballoon is immersed in the 0.5mol/L NaCl solution of 1mg/mLPSS same standing adsorption centrifuge washing.After adsorbing one deck PAH again, microballoon is immersed in the CdTe that mercaptoacetic acid is modified
540In (emission spectrum is at 540nm) aqueous solution, standing adsorption is centrifuge washing after 60 minutes.
3 repeat above-mentioned steps.After adsorbing three strata electrolyte, adsorb second layer CdTe again
540By that analogy, adsorbing three layers of CdTe
540After, adsorb four strata electrolyte again at outermost quantum dot, obtain a kind of composite coding microballoon.
4 equally with the silicon dioxide microsphere of 540nm coding, adsorbing three layers of CdTe
540After, continue six layers of CdTe of parcel again
580, adsorb four strata electrolyte again at outermost quantum dot, obtain another coding microball.
5 equally with the silicon dioxide microsphere of 540nm coding, adsorbing three layers of CdTe
540After, continue three layers of CdTe of parcel again
580, and then adsorb three layers of CdTe
620, adsorb four strata electrolyte again at outermost quantum dot, obtain another coding microball.
6 will wrap the coding microball mixing of being crossed, and obtain three kinds of not homospecific composite coding microballoons.Be the 540nm that is encoded to of photon crystal micro-ball, quantum-dot coding is respectively CdTe
540=3, CdTe
540: CdTe
560=3: 6, CdTe
540: CdTe
580: CdTe
620=3: 3: 3 photon crystal composite encoding microsphere.
Claims (10)
1. photon crystal composite encoding microsphere, it is characterized in that quantum dot being wrapped in the photon crystal micro-ball surface of coding with the method for layer assembly, by wrapping up the different quantum dots and the different quantum dot numbers of plies, make quantum-dot coding combine with the photon crystal micro-ball coding, encoding amount increases greatly.
2. photon crystal composite encoding microsphere according to claim 1 is characterized in that described photon crystal micro-ball is the opal structural photonic crystal of the orderly self assembly of colloidal particle, or the photonic crystal of counter opal structure.
3. a kind of photon crystal composite encoding microsphere according to claim 1 is characterized in that described quantum dot is cadmium sulfide CdS, cadmium selenide CdSe, cadmium telluride CdTe or nucleocapsid quantum dot.
4. a kind of photon crystal composite encoding microsphere according to claim 3 is characterized in that described nucleocapsid quantum dot is a zinc sulphide bag cadmium selenide CdSe@ZnS quantum dot.
5. according to claim 1,2 described a kind of photon crystal composite encoding microspheres, the photon crystal micro-ball that it is characterized in that described coding are a kind of in polystyrene, polymethylmethacrylate, polyacrylonitrile, tygon, polypropylene, the polyethyl acrylate; Or be silicon dioxide microsphere or titanium dioxide microballoon sphere.
6. the preparation method of a kind as a kind of photon crystal composite encoding microsphere according to claim 1 is characterized in that the preparation method specifically may further comprise the steps:
1) chooses the photon crystal micro-ball of different coding, it is carried out surface treatment, obtain electronegative photonic crystal coding microball; Wherein photon crystal micro-ball is the polyvinyls microballoon or is inorganic photon crystal micro-ball;
2) utilize the method for absorption layer by layer, earlier at the positive polyelectrolyte of photon crystal micro-ball surface adsorption, the negative polyelectrolyte of absorption then, adsorb positive polyelectrolyte again after, the quantum dot of different characteristic emission peak is adsorbed on the microballoon outside surface;
3) to adsorb the quantum dot of the different numbers of plies or variety classes size, layer with layer between separate with three strata electrolyte;
4) continue to adsorb four strata electrolyte again at outermost quantum dot, so that detect.
7. a kind of photon crystal composite encoding microsphere according to claim 6 is characterized in that described positive polyelectrolyte can be polypropylene amine hydrochloride PAH, also can be polyethyleneimine PEI; Used negative polyelectrolyte is kayexalate PSS, also can be polyacrylic acid PAA.
8. the preparation method of a kind of photon crystal composite encoding microsphere according to claim 6 is characterized in that described quantum-dot coding is the feature emission peak of quantum dot, and the feature emission peak that utilizes different quantum dots is as different quantum-dot codings.
9. the preparation method of a kind of photon crystal composite encoding microsphere according to claim 6, it is characterized in that described quantum-dot coding is the intensity of the feature emission peak of quantum dot, control the intensity of its feature emission peak by the quantum dot that adsorbs the different numbers of plies, the intensity of utilizing the different characteristic emission peak is as different quantum-dot codings.
10. the preparation method of a kind of photon crystal composite encoding microsphere according to claim 6, what it is characterized in that described quantum dot is encoded to its feature emission peak and its feature emission peak combination of strength, utilizes the absorption variety classes and the different numbers of plies to control the intensity of feature emission peak He its feature emission peak of quantum dot.
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