A three-fluorescence-emitting molecularly imprinted sensor and its preparation and application
A technology of molecular imprinting and fluorescence emission, which is applied in the field of analytical chemistry and rapid detection, can solve the problems of inability to accurately visualize the detection target, complex optimization process of emission peak intensity ratio, narrow range of fluorescence color change, etc., to achieve rich fluorescence color change , reduce experiment cost and shorten experiment period
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Embodiment 1
[0036] Preparation of three fluorescence-emitting molecularly imprinted sensors:
[0037] (1) Preparation of bovine hemoglobin-imprinted microspheres, taking blue fluorescent bovine hemoglobin-imprinted microspheres as an example, see figure 1 :
[0038] To 20 mL of phosphate buffer (0.01 M, pH 7.0), add 10 mg of silica microspheres with a particle size of approximately 79 nm, 10 mg of bovine hemoglobin, and 35 μL of 3-aminopropyltriethoxysilane (APTES), and stir for 1 Hours later, 20 mg of blue fluorescent 7-hydroxycoumarin was added, and after mixing, 56 μL of ethyl orthosilicate (TEOS) and 56 μL of ammonia (NH) were added. 3 ·H 2 O), the sol-gel polymerization was carried out in the dark for 10-12 h. After the reaction was completed, the product was precipitated by centrifugation (8000 rpm, 5 minutes), the supernatant was discarded, and the bovine hemoglobin in the precipitate was eluted with 0.5% Triton X-100 to obtain a blue fluorescent bovine hemoglobin blot with a co...
Embodiment 2
[0045] The red fluorescent bovine hemoglobin-imprinted microspheres (r-MIPs) dispersion liquid, blue fluorescent bovine hemoglobin-imprinted microspheres (b-MIPs) dispersion liquid and green fluorescent bovine hemoglobin-imprinted microspheres (g-MIPs) dispersion liquid obtained above were determined according to certain conditions. The sensor was obtained by mixing the ratio in phosphate buffer (0.01 M, pH 7.0); the total volume of phosphate buffer (0.01 M, pH 7.0) was controlled at 1 mL, and the amount of red bovine hemoglobin blotting microspheres (r-MIPs) was fixed. is 5% (volume fraction), the dosage of blue bovine hemoglobin-imprinted microspheres (b-MIPs) is fixed at 6% (volume fraction), and the dosage of green bovine hemoglobin-imprinted microspheres (g-MIPs) is 0%, 3%, 5%, 7%, 9%, 11%, 13% and 15%; then different amounts of bovine hemoglobin were added to the above sensor to achieve final concentrations of 0, 0.025, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75 , 1, 2 and 3 μM; ...
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 solution according to a certain ratio, that is, different sensors with different selections and different ratios are obtained; wherein the red-green-blue three fluorescence emission molecular imprinting r-MIPs in the sensor: g-MIPs: b-MIPs=5%:11%:6% (volume percentage), red-green-blue three fluorescence emission r-NIPs: g-NIPs: b-NIPs in the non-imprinted sensor =5%:11%:6% (volume percentage), r-MIPs: g-MIPs in the red-green dual fluorescence emission molecular imprinting sensor =5%:11% (volume percentage), red-blue dual fluorescence emission molecular imprinting r-MIPs in the sensor: b-MIPs=5%:6% (volume percentage), g-MIPs in the green-blue dual fluorescence emission molecular imprint sensor: b-MIPs=11%:6% (volume percentage); and, The total volume of phosphate buffer (0.01 M, pH 7.0) in each of the ab...
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