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Tri-fluorescent emission molecularly imprinted sensor and preparation method and application thereof

A molecular imprinting and fluorescence emission technology, applied in the field of analytical chemistry and rapid detection, can solve the problems of inability to accurately visualize the detection target, complicated optimization process of emission peak intensity ratio, narrow fluorescence color variation range, etc., and achieve rich fluorescence color variation. , The effect of reducing the experimental cost and shortening the experimental period

Active Publication Date: 2019-04-19
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are two main deficiencies in the ratiometric fluorescent molecular imprinting sensor: (1) The traditional ratiometric fluorescent molecular imprinting sensor only has two emission peaks, and the fluorescence color range provided when detecting the target is narrow, and cannot accurately visualize the detection target. (2) The preparation of ratiometric fluorescent molecularly imprinted sensors often needs to optimize the fluorescence intensities of different emission peaks, and select the appropriate emission peak intensity ratio to achieve the best visual detection effect, but the emission peaks of traditional ratiometric fluorescent molecularly imprinted sensors The strength ratio optimization process is complex and tedious

Method used

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  • Tri-fluorescent emission molecularly imprinted sensor and preparation method and application thereof
  • Tri-fluorescent emission molecularly imprinted sensor and preparation method and application thereof
  • Tri-fluorescent emission molecularly imprinted sensor and preparation method and application thereof

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Embodiment 1

[0036] Preparation of three fluorescence-emitting molecularly imprinted sensors:

[0037] (1) Prepare bovine hemoglobin imprinted microspheres, take blue fluorescent bovine hemoglobin imprinted microspheres as an example, see figure 1 :

[0038] Add 10 mg of silica microspheres with a particle size of about 79 nm, 10 mg of bovine hemoglobin and 35 μL of 3-aminopropyltriethoxysilane (APTES) into 20 mL of phosphate buffer (0.01 M, pH 7.0), and stir for 1 After 1 hour, add 20 mg of blue fluorescent 7-hydroxycoumarin, mix well and continue to add 56 μL tetraethyl orthosilicate (TEOS) and 56 μL ammonia water (NH 3 ·H 2 O), the sol-gel polymerization was carried out in the dark for 10-12 hours. After the reaction is completed, the product is precipitated by centrifugation (8000rpm, 5 minutes), the supernatant is discarded, and the bovine hemoglobin in the precipitate is eluted with 0.5% Triton X-100 to obtain a blue fluorescent bovine hemoglobin blot with a nucleocapsid structure...

Embodiment 2

[0045] The above obtained red fluorescent bovine hemoglobin imprinted microspheres (r-MIPs) dispersion, blue fluorescent bovine hemoglobin imprinted microspheres (b-MIPs) dispersion and green fluorescent bovine hemoglobin imprinted microspheres (g-MIPs) dispersion were The sensor was obtained by mixing in phosphate buffer (0.01 M, pH 7.0) in proportion; the total volume of phosphate buffer (0.01 M, pH 7.0) was controlled at 1 mL, and the amount of red bovine hemoglobin imprinted microspheres (r-MIPs) was fixed 5% (volume fraction), the amount of blue bovine hemoglobin imprinted microspheres (b-MIPs) was fixed at 6% (volume fraction), and the amount of green bovine hemoglobin imprinted microspheres (g-MIPs) was 0%, 3%, respectively. 5%, 7%, 9%, 11%, 13% and 15%; then add different amounts of bovine hemoglobin to the above sensor to make the final concentration 0, 0.025, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75 , 1, 2 and 3 μM; add bovine hemoglobin and mix well, react for 8 minutes, a...

Embodiment 3

[0048] Different sensors are prepared according to the sensor preparation method described above. Specifically, the different microspheres obtained above are mixed in a buffer in a certain proportion to obtain different sensors with different selections and different proportions; among them, red-green-blue three fluorescent emission molecular imprints r-MIPs in the sensor: g-MIPs: b-MIPs=5%:11%:6% (volume percentage), red-green-blue three fluorescent emission non-imprinted sensor r-NIPs: g-NIPs: b-NIPs =5%:11%:6% (volume percentage), r-MIPs: g-MIPs in the red-green dual fluorescent emission molecularly imprinted sensor =5%:11% (volume percentage), red-blue dual fluorescent emission molecularly imprinted sensor r-MIPs in the sensor: b-MIPs=5%:6% (volume percentage), g-MIPs in the green-blue dual fluorescence emission molecularly imprinted sensor: b-MIPs=11%:6% (volume percentage); and, The total volume of phosphate buffer (0.01 M, pH 7.0) in each of the above sensors was contro...

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Abstract

The invention belongs to the field of analytical chemistry and quick detection, and particularly relates to a red-green-blue tri-fluorescent emission molecularly imprinted sensor based on a post-imprinting mixing method and a preparation method and application thereof in accurately and visually detecting hemoglobin. The red-green-blue tri-fluorescent emission molecularly imprinted sensor is obtained by mixing red fluorescent bovine hemoglobin imprinted microspheres, green fluorescent bovine hemoglobin imprinted microspheres and blue fluorescent bovine hemoglobin imprinted microspheres throughthe post-imprinting mixing method. The sensor prepared by the method can detect bovine hemoglobin in a high-sensitivity, high-selectivity and self-correcting manner, and the sensor is wider and richerthan conventional dual-fluorescent emission molecularly imprinted sensors in fluorescent color changing range and can visually detect a target more accurately.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and rapid detection, and in particular relates to a red-green-blue three-fluorescent emission molecular imprint sensor based on a post-imprint mixing method, its preparation and its application in accurate and visual detection of bovine hemoglobin. Background technique [0002] Nowadays, rapid visual detection plays an important role in environmental detection, food safety, clinical diagnosis and other fields. Rapid visual detection of a large number of samples to be tested, combined with accurate detection of suspicious samples with large instruments (such as high performance liquid chromatography) often constitute a complete detection process, which not only saves money and time spent on detection, but also ensures the reliability of detection results. Therefore, rapid visual detection methods are often required to have high sensitivity and high selectivity. [0003] The ratiometric fluoresc...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 李金花杨倩陈令新于祖海
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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