Functional nano-rare earth fluorescent micro particle and its preparation and application
A technology of nano-rare earth and fluorescent particles, which is applied in the field of nano-rare-earth fluorescent materials, can solve the problems of unsuitable nano-fluorescent particles, low measurement sensitivity, weak fluorescence, etc., and achieve the effects of improving measurement sensitivity, solving fluorescence bleaching, and enhancing photostability
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Embodiment 1
[0048] Contains 4'-(2-thienyl)-2,2':6',2"-biteropyridine-6,6"-dimethylaminetetraacetic acid europium (TTTA-Eu for short) 3+ ) Preparation of nano fluorescent particles:
[0049] (1) TTTA-Eu 3+ Preparation of complexes
[0050] 411 mg of TTTA·H 2 O was dissolved in 6ml of water, with solid NaHCO 3 Adjust the pH to 6.5. Then 116 mg of Eu 2 o 3 After heating and dissolving with dilute hydrochloric acid, add it to TTTA aqueous solution, and use solid NaHCO 3 Adjust the pH to 6.5, stir at room temperature for 2 hours, remove the precipitate by filtration, add 300ml of acetone, stir at room temperature for 1 hour, collect the precipitate by filtration, dry in vacuo, wash the dry solid with dry methanol, evaporate the filtrate to dryness under reduced pressure, and dry in vacuo. 470mg of TTTA-Eu can be obtained 3+ Complex (yield 67.7%).
[0051] (2) Contains europium complex TTTA-Eu 3+ Preparation of fluorescent nanoparticles
[0052] The europium fluorescent complex TTTA...
Embodiment 2
[0062] Use TTTA-Eu 3+ Determination of alpha-fetoprotein (AFP) in human serum by time-resolved fluorescent immunoassay with complex LFNP as marker
[0063] (1) Use nano-rare earth fluorescent particles to label streptavidin (SA)
[0064] Disperse the nanospheres to 2mol / L Na 2 CO 3 In the aqueous solution, after 20 minutes of ultrasonic activation, add the acetonitrile solution dissolved in CNBr, react at room temperature for 2-10 minutes, centrifuge to collect the precipitate, and use ice water and 0.1mol / L NaHCO for the precipitate 3 (pH=8.5) buffer solution was washed twice, and then the activated nanoparticles were added to the aqueous solution containing bovine serum albumin (abbreviated as BSA), and reacted overnight, with 0.1mol / L NaHCO 3(pH=8.5) After washing with buffer solution, add SA and glutaraldehyde to react for 24 hours, then add NaBH 4 Reduction, react at room temperature for 30 minutes, wash several times and suspend in solution containing 0.2% BSA-0.1% N...
Embodiment 3
[0072] Contains N, N, N 1 , N 1 -[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-(2"-phenyl)pyridine]terbium tetraacetate (referred to as BPTA-Tb 3+ ) preparation of fluorescent nanoparticles:
[0073] (1) BPTA-Tb 3+ Preparation of complexes
[0074] Dissolve 192 mg of BPTA (see Document 7 for the synthetic method: Yuan J, Wang G, Majima K, Matsumoto K.nal.Chem.2001, 73, 1869-1876) in 4 ml of water, and use NaHCO 3 Adjust the pH to 6.5. Then 120mg of TbCl 3 The solution dissolved in 2ml of water was added to the BPTA aqueous solution, with NaHCO 3 Adjust the pH to 6.5, stir at room temperature for 2 hours, remove the precipitate by filtration, add 300ml of acetone to the filtrate, stir at room temperature for 1 hour, collect the precipitate by filtration, dry in vacuo, wash the dry solid with dry methanol, evaporate the filtrate to dryness in vacuo After drying, 198mg of BPTA-Tb can be obtained 3+ Complex (72.9% yield).
[0075] (2) Contains BPTA-Tb 3+ Preparation of Fluores...
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