CN115011345A - Preparation method of bicolor quantum dot polystyrene microspheres - Google Patents
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Abstract
The invention relates to a preparation method of a bicolor quantum dot polystyrene microsphere, which is characterized in that quantum dot polystyrene microspheres with different proportions are prepared by controlling experimental materials by a PSMA (patterned sapphire plasma enhanced chemical vapor deposition) medium method. The bicolor quantum dot polystyrene microsphere synthesized by the method has the characteristics of good uniformity, stable performance and stronger bicolor fluorescent signals. The mesoporous polystyrene is used as a matrix, and the quantum dots are connected to the interior of the microsphere by a poly (styrene-maleic anhydride) mediated method, so that a stable structure can be obtained. The prepared bicolor quantum dot polystyrene microsphere has good uniformity, physical and chemical stability and fluorescence luminescent performance. The preparation method disclosed by the invention is simple in process and strong in operability, and can further meet the requirements of production and application.
Description
Technical Field
The invention relates to a preparation method of a bi-color quantum dot polystyrene microsphere, in particular to a preparation technology for simultaneously connecting oil-soluble cadmium quantum dots CdSe/ZnS with two different wavelengths by using mesoporous polystyrene as a matrix. The invention belongs to the field of nano biological medicine materials.
Background
At present, the global burden of cancer morbidity and mortality is increasing, and cancer has surpassed high-mortality chronic diseases such as cardiovascular diseases (such as stroke and coronary heart disease) in the cause-of-death ranking of many countries, which reflects both the acceleration of the aging process of the population and the increase of exposure of cancer-related risk factors. Cancerous tumors can metastasize a small number of tumor cells into their direct vasculature, and these cells then enter the circulatory system and subsequently spread further to distant organs, often by the time they are found to have reached an advanced stage in the cancer. Therefore, how to accurately detect cancer at an early stage and obtain timely treatment is a crucial problem. This not only reduces the economic burden of the patient, but also improves the therapeutic effect and reduces the cancer mortality. Aiming at the clinical requirement of early diagnosis of cancer, the project establishes a novel detection method for measuring exosome protein and nucleic acid by taking a liquid-phase suspension chip technology as a means. The main idea is that the space regulation and control quantum dots are distributed inside and outside the carrier to prepare the multicolor fluorescent microsphere with stable fluorescence performance. The colloidal stability, the optical property stability and the nonspecific adsorption condition of the microsphere after the surface ligand modification are explored, and the coding microsphere with good specificity is prepared. The multicolor quantum dot fluorescent microspheres can be applied to a subsequent multi-target combined detection platform at a molecular level, so that capture, identification and separation of different types of exosomes are realized, and high-sensitivity determination of exosome proteins and nucleic acids is realized.
The traditional fluorescent microspheres mainly comprise the following fluorescent coding elements: fluorescent dye (Chinese invention patent: a preparation method of rose bengal B/polystyrene/silicon dioxide fluorescent microspheres, publication No. CN 201811287084.6; Chinese invention patent: a near-infrared fluorescent microspheres with an emission peak value of not less than 1300nm and preparation and application thereof, publication No. CN 201911055089.0; Chinese invention patent: a preparation method of fluorescent microspheres marked by mixed fluorescent dye, publication No. CN 201910227942.6), rare earth elements (Chinese invention patent: a preparation method of rare earth fluorescent microspheres, publication No. CN 201710007690.7; Chinese invention patent: a rare earth complex fluorescent microspheres, preparation method and application thereof, publication No. CN 202110880268.9), carbon quantum dots (Chinese patent: carbon quantum dot fluorescent microsphere standard substance and preparation method thereof, publication number: CN 201911409568.8) and optical coding elements developed at present are considered to be quantum dot materials most suitable for optical coding.
In the planar array and the suspended array of the mainstream multiplexing array, the quantum dot coding microsphere multiplexing suspended array has the advantages of rapid combination dynamics, flexibility in target selection, well-controlled combination conditions and the like, and can provide data with higher quality. The kit is widely applied to the fields of multi-index combined diagnosis or detection (Chinese invention patent: a detection method and a detection kit based on fluorescent coding magnetic beads, publication number: CN 201910477217.4; Chinese invention patent: coding microspheres for biomolecule screening or detection, preparation method and application thereof, publication number: CN 201610935639.8; Chinese invention patent: PCT/IL-6 dual detection kit based on quantum dot fluorescent microspheres, publication number: CN 202110988231.8; Chinese invention patent: a combined detection kit for bladder cancer determination and preparation method thereof, publication number: CN 201810418743.9), DNA hybridization, spectrum correction and immunoassay (Chinese invention patent: quantum dot coding fluorescence immunoassay instrument, publication number: CN 201010165974.7).
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation method of a bicolor quantum dot polystyrene microsphere. The method brings convenience to joint detection and is beneficial to the subsequent establishment of a systematic 3D coding library.
The purpose of the invention is realized by the following scheme: a preparation method of bi-color quantum dot polystyrene microspheres takes mesoporous polystyrene Microspheres (MPS) as a matrix, and simultaneously connects green and red CdSe/ZnS quantum dots with the wavelengths of 538nm and 623nm by using a poly (styrene-maleic anhydride) mediated method (PSMA) to prepare the quantum dot polystyrene microspheres (Q-MPS), which comprises the following steps:
it is characterized in that
(1) Preparing NaOH alkali liquor with the molar concentration of 0.05 mol/L; weighing 5-8 mg of MPS powder; in addition, 1-3 mg of PSMA is weighed, dispersed in 90 mu l of chloroform and subjected to ultrasonic treatment, and the materials are prepared and then placed for later use;
(2) adding 60 uL of green and red quantum dot chloroform solution with a certain volume ratio of 5mg/mL into the PSMA chloroform solution, controlling the total volume to be 150 muL, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS, and performing ultrasonic treatment for 15 min; adding 500 μ L of prepared 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water, immediately obtaining the fluorescent quantum dot coding microspheres, dispersing the particles in water, and storing the particles in a dark place.
The Q-MPS obtained by the method has good fluorescent effect, the particles are uniformly dispersed, the interference between double-color fluorescence is small, and the two-color fluorescence shows obvious peak values.
The polystyrene microsphere is a mesoporous polystyrene microsphere.
The two-color quantum dots can be CdSe/ZnS quantum dots with the wavelength of 538nm and 623 nm.
The volume ratio of the two-color quantum dots is 50: 1-1: 50.
The invention has the advantages that:
(1) the invention takes mesoporous polystyrene as a matrix, and quantum dots are connected to the inside of the microsphere by a poly (styrene-maleic anhydride) mediated method, so that a stable structure can be obtained.
(2) The prepared bicolor quantum dot polystyrene microsphere has good uniformity, physical and chemical stability and fluorescent light-emitting performance.
(3) The preparation method disclosed by the invention is simple in process and strong in operability, and can further meet the requirements of production and application. Drawings
FIG. 1 is a photograph of an ultraviolet lamp irradiation of the quantum dot polystyrene microsphere prepared in example 1;
FIG. 2 is a fluorescence intensity curve of the quantum dot polystyrene microsphere prepared in example 1;
FIG. 3 is a particle size distribution diagram of the quantum dot polystyrene microspheres prepared in example 2;
FIG. 4 is a fluorescence intensity curve of the quantum dot polystyrene microsphere prepared in example 2;
FIG. 5 is a scanning electron microscope image of the quantum dot polystyrene microspheres prepared in example 3;
FIG. 6 is a transmission electron microscope image of the quantum dot polystyrene microsphere prepared in example 3.
Detailed Description
The technical solution of the present invention is further described below by specific examples. The following examples are further illustrative of the present invention and do not limit the scope of the present invention.
Example 1
A bi-color quantum dot polystyrene microsphere (Q-MPS) is prepared by taking a mesoporous polystyrene Microsphere (MPS) as a matrix, simultaneously connecting green and red CdSe/ZnS quantum dots with the wavelengths of 538nm and 623nm by using a poly (styrene-maleic anhydride) mediated method (PSMA), and comprises the following steps:
(1) preparing 7.5ml of NaOH alkaline solution with the molar concentration of 0.05 mol/L; weighing 15 parts of 6.85 mgMPS powder; and additionally weighing 15 parts of 2 mg PSMA, dispersing in 90 mu l chloroform and carrying out ultrasonic treatment to obtain each chloroform solution for later use; subsequently, the process of the present invention,
(2) adding green and red quantum dot chloroform solutions (numbered as table 1 below) with volume of 60 uL and volume of 5mg/mL into PSMA chloroform solution, controlling the total volume to be 150 μ L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS chloroform solution, and performing ultrasonic treatment for 15 min; adding 500 μ L of 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; then, the reaction mixture was centrifuged and washed with ultrapure water 3 times, the numbers of which are shown in table 1 below, to obtain fluorescent quantum dot-encoded microspheres, and the particles were dispersed in water and stored in the dark.
Fig. 1 and fig. 2 show the ultraviolet lamp irradiation photograph and fluorescence curve of the green-red two-color quantum dot polystyrene microsphere in different proportions of each group of quantum dot polystyrene microspheres in example 1, respectively, and it can be seen from the figure that when the left image, i.e. green light, is in large proportion, and the proportion is 1:1 to 10:1, the intensity of red and green light is equivalent, and the ultraviolet lamp image presents yellow and orange excessive colors, which is a more ideal state.
Example 2
A bicolor quantum dot polystyrene microsphere is prepared by the following steps similar to the steps of example 1:
(1) preparing 5ml of NaOH alkaline liquor with the molar concentration of 0.05 mol/L; weighing 10 parts of 6.85 mgMPS powder; then 10 parts of 2 mg PSMA is weighed, dispersed in 90 mul chloroform and subjected to ultrasonic treatment, and the materials are prepared and placed for standby; followed by
(2) Adding green and red quantum dot chloroform solutions with different volumes of 5mg/mL into the PSMA chloroform solution, wherein the numbers are shown in the following table 2, the total volume is controlled to be 150 mu L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS, and performing ultrasonic treatment for 15 min; adding 500 μ L of prepared 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; then, the reaction mixture was centrifuged and washed with ultrapure water 3 times to obtain fluorescent quantum dot-encoded microspheres numbered as in table 2 below, and the particles were dispersed in water and stored in the dark.
Since the microspheres show more performance in the ratio of 10:1 to 1:1, the ratio is refined to 10 samples of 10:1,9:1,8:1,7:1,6:1,5:1,4:1,3:1,2:1 and 1: 1.
Fig. 3 is a particle size image obtained by performing a micrometer particle size analyzer test on green-red dual-color polystyrene quantum dots Q-MPS in each proportion in example 2, and it can be seen that the particle sizes of materials in different proportions are similar, and all have an obvious peak value around 5 micrometers, and the image preliminarily determines that the dual-color coded microspheres exhibit good uniformity. FIG. 4 shows fluorescence intensities of quantum dot microspheres of example 2 at different ratios.
It can be seen from table 3 that the ratio of the total fluorescence intensity area shows an increasing trend with the increase of the green light ratio in the number, indicating that the interference between the two quantum dots is small.
Example 3
A bicolor quantum dot polystyrene microsphere is prepared by the following steps similar to the steps of example 1:
(1) preparing 1.5ml of NaOH alkaline solution with the molar concentration of 0.05 mol/L; weighing 3 parts of 6.85 mgMPS powder; then 3 parts of 2 mg PSMA is weighed, dispersed in 90 mul chloroform and subjected to ultrasonic treatment, and the materials are prepared and placed for standby; subsequently, the process of the present invention,
(2) adding green and red quantum dot chloroform solutions (numbered as table 4 below) of 5mg/mL in different volumes into the PSMA chloroform solution, controlling the total volume to be 150 μ L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS, and performing ultrasonic treatment for 15 min; adding 500 μ L of prepared 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water, wherein the serial number is shown in the following table 3, immediately obtaining the fluorescent quantum dot coding microspheres, dispersing the particles in water, and storing the particles in a dark place.
FIG. 5 is a scanning electron microscope image of the two-color quantum dot polystyrene microspheres of 1:1,3:1,5:1 prepared in example 3, and it is obvious from the 700 times scanning electron microscope image that most of the microspheres exist independently and have good uniformity. From the images of 6k, 15k times, it can be seen that the microspheres still maintain regular spherical shapes and have more obvious mesopores on the surface.
FIG. 6 is a TEM image of 1:1,5:1 two-color quantum dot polystyrene microspheres prepared in example 3. The transmission electron microscope images show that the quantum dots in all proportions are similar in appearance, and the high-resolution images show that a large number of quantum dots with obvious lattice structures exist inside the high-resolution images, and the sizes of the two quantum dots are not greatly different.
Claims (5)
1. A preparation method of bi-color quantum dot polystyrene microspheres is characterized in that mesoporous polystyrene Microspheres (MPS) are used as a matrix, a poly (styrene-maleic anhydride) dielectric method (PSMA) is used, green and red CdSe/ZnS quantum dots with the wavelengths of 538nm and 623nm are connected at the same time, and quantum dot polystyrene microspheres (Q-MPS) are prepared, and the preparation method comprises the following steps:
(1) preparing NaOH alkali liquor with the molar concentration of 0.05 mol/L; weighing 5-8 mg of MPS powder and 1-3 mg of PSMA, respectively dispersing in 90 mu l of chloroform, and carrying out ultrasonic treatment to obtain chloroform solutions for later use;
(2) adding green and red quantum dot chloroform solutions with volume and volume of 60 uL and 5mg/mL into the PSMA chloroform solution, wherein the volume ratio of the green to red quantum dot chloroform solutions is 50:1 to 1:50, controlling the total volume to be 150 mu L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of perovskite Quantum Dots (QDs) and PSMA into MPS chloroform solution, and carrying out ultrasonic treatment for 15 min; adding 500 μ L of 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then, centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water, immediately obtaining the two-color quantum dot polystyrene microsphere fluorescent quantum dot coding microsphere, dispersing the particles in water, and storing the particles in a dark place.
2. The preparation method of the bi-color quantum dot polystyrene microsphere of claim 1, wherein the volume ratio of the bi-color quantum dots is 50: 1-1: 50.
3. The preparation method of the bi-color quantum dot polystyrene microsphere according to claim 1 or 2, which is characterized by comprising the following steps:
(1) preparing 7.5ml of NaOH alkaline solution with the molar concentration of 0.05 mol/L; weighing 15 parts of 6.85 mg MPS powder; in addition, 15 parts of 2 mg PSMA is weighed, dispersed in 90 mul chloroform and subjected to ultrasonic treatment to obtain each chloroform solution for later use; subsequently, the air conditioner is operated to,
(2) adding 15 parts of green and red quantum dot chloroform solutions with different volume ratios, wherein the volume ratio of the green and red CdSe/ZnS quantum dots is 60 uL, and the volume ratio of the green and red CdSe/ZnS quantum dots is respectively as follows: 50: 1. 30: 1. 20: 1. 10: 1. 5: 1. 3: 1. 2: 1. 1: 1. 1: 2. 1: 3. 1: 5. 1: 10. 1: 20. 1: 30 and 1:50, controlling the total volume to be 150 mu L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS chloroform solution, and performing ultrasonic treatment for 15 min; adding 500 μ L of 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water to obtain a bicolor fluorescent quantum dot coding microsphere sample, dispersing the particles in water, and storing the particles in a dark place.
4. The preparation method of the bi-color quantum dot polystyrene microsphere according to claim 1 or 2, which is characterized by comprising the following steps:
(1) preparing 5ml of NaOH alkaline liquor with the molar concentration of 0.05 mol/L; weighing 10 parts of 6.85 mgMPS powder; then 10 parts of 2 mg PSMA is weighed, dispersed in 90 mul chloroform and subjected to ultrasonic treatment, and the materials are prepared and placed for standby; followed by
(2) Adding 10 parts of green and red quantum dot chloroform solutions with different volume ratios, the volume ratio of which is 60 uL, into PSMA chloroform solution, wherein the volume ratio of green and red CdSe/ZnS quantum dots is respectively as follows: 10: 1. 9: 1. 8: 1. 7: 1. 6: 1. 5: 1. 4: 1. 3: 1. 2:1 and 1:1, controlling the total volume to be 150 mu L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS, and performing ultrasonic treatment for 15 min; adding 500 μ L of prepared 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then, centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water, obtaining 10 two-color fluorescent quantum dot coding microsphere samples with different numbers, dispersing the particles in water, and storing the particles in a dark place.
5. The preparation method of the bi-color quantum dot polystyrene microsphere according to claim 1 or 2, which is characterized by comprising the following steps:
(1) preparing 1.5ml of NaOH alkaline solution with the molar concentration of 0.05 mol/L; weighing 3 parts of 6.85 mgMPS powder; then 3 parts of 2 mg PSMA is weighed, dispersed in 90 mul chloroform and subjected to ultrasonic treatment, and the materials are prepared and placed for standby; subsequently, the process of the present invention,
(2) adding 3 parts of green and red quantum dot chloroform solutions with different volume ratios, wherein the volume ratio of the green and red CdSe/ZnS quantum dots is 60 uL, and the volume ratio of the green and red CdSe/ZnS quantum dots is respectively as follows: 1: 1. 3:1 and 5:1, controlling the total volume to be 150 mu L, and performing ultrasonic dispersion; transferring the mixed chloroform dispersion liquid of QDs and PSMA into prepared MPS, and performing ultrasonic treatment for 15 min; adding 500 μ L of prepared 0.05mol/L NaOH alkaline solution, and performing ultrasonic treatment for 10 min; and then centrifuging the reaction mixed solution, washing the reaction mixed solution for 3 times by using ultrapure water, immediately obtaining 3 bicolor fluorescent quantum dot coding microsphere samples, dispersing the particles in water, and storing the particles in a dark place.
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Citations (3)
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| KR20120101242A (en) * | 2011-02-28 | 2012-09-13 | 고려대학교 산학협력단 | Quantum dot encapsulated ps-psma nanofibers and a use of the same |
| CN105080439A (en) * | 2015-06-26 | 2015-11-25 | 上海交通大学 | Microspheres with high fluorescence intensity and preparation method for microspheres |
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| CN105080439A (en) * | 2015-06-26 | 2015-11-25 | 上海交通大学 | Microspheres with high fluorescence intensity and preparation method for microspheres |
| CN110776916A (en) * | 2019-09-20 | 2020-02-11 | 浙江工业大学 | Quantum dot dual-emission-ratio fluorescent probe and preparation method and application thereof |
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