Rare-earth nano luninous particle based on fluorescent energy transfer principle and its preparing method

Abstract

The invention relates to emitting rare earth nm particle based on fluorescence energy transferring principle and the method to manufacture, and new style rare earth nm particle manufacturing and the surface decorating used for testing biology molecule mark and tracing. The rare earth nm particle is made up from emitting rare earth complexes as the core and covering SiO2. The rare earth complex is made up from organic ligand joining with a sensitization molecule and rare earth ion. The rare earth nm particle marker has good water solubility and long fluorescence lifetime. The surface of the particle could covalence joining plural probe molecule that could be used to testing or tracing the biology molecule.

Description

technical field

[0001] The invention relates to a rare earth nano particle emitting light based on the principle of fluorescence energy transfer and a preparation method thereof, which is mainly used for label detection and tracing of biomolecules, and belongs to the technical field of preparation of biomolecular nano-markers. Background technique

[0002] Fluorescent labeling and detection technology has a wide range of applications in life sciences, medicine, immunology and other fields. However, traditional fluorescence analysis is susceptible to interference from nonspecific fluorescence. Most of the currently used fluorescent markers are various organic dye molecules. Since the excitation wavelength is close to the emission wavelength and the Stokes shift is small (20-30nm), the detection is easily interfered by the excitation light. In addition, the background fluorescence and scattered light almost cover the entire range of the fluorescence emission spectrum (350-600...

Examples

Embodiment 1

[0026] Embodiment 1: Preparation of 7-amino-4-methyl-2-quinolinone-DTPA-Tb nanoparticles modified by oligonucleotide probe:

[0027] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: dissolve equimolar amounts of 7-amino-4-methyl-2-quinolinone and diethyltriaminepentaacetic dianhydride in anhydrous In dimethylformamide (DMF), add an appropriate amount of anhydrous triethylamine to the DMF solution of diethyltriaminepentaacetic dianhydride and add 7-amino-4-methyl-2-quinolinone with dropwise stirring DMF solution, reacted for 2 hours under the condition of avoiding light and stirring.

[0028] Preparation of rare earth complexes; equimolar amounts of rare earth Tb 3+ Aqueous solution (Tb 3+ Nitrate or hydrochloride) was mixed with the above-mentioned conjugate solution for 10 minutes.

[0029] Preparation of rare earth nanoparticles: Mix cyclohexanone, n-hexanol, Triton X-100 (v / v, 4:1:1) and stir evenly, add an appropriate amoun...

Embodiment 2

[0031] Embodiment 2: Preparation of aminofluorescein-DTPA-Yb nanoparticles:

[0032] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: Dissolve equimolar amounts of aminofluorescein and diethyltriaminepentaacetic dianhydride in anhydrous dimethylformamide (DMF), Add an appropriate amount of anhydrous triethylamine to the DMF solution of diethyltriaminepentaacetic dianhydride, stir the DMF solution of aminofluorescein dropwise, and react for 2 hours under the condition of avoiding light and stirring.

[0033] Preparation of rare earth complexes: equimolar amounts of rare earth Yb 3+ Aqueous solution (Yb 3+ hydrochloride) was mixed with the above conjugate solution for 10 minutes.

[0034] Preparation of rare earth nanoparticles: Mix cyclohexanone, n-hexanol, Triton X-100 (v / v, 4:1:1) and stir evenly, add an appropriate amount of rare earth complex solution (water phase), and fully stir to form After the water microemulsion, add t...

Embodiment 3

[0035] Example 3: Preparation of 7-amino-4-methyl-2-quinolinone-TTHA-Eu nanoparticles:

[0036] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: dissolve a certain molar amount of triethylenetetraminehexaacetic acid (TTHA) in anhydrous dimethylformamide (DMF), and add a small amount of anhydrous Triethylamine was reacted for 12 hours under stirring conditions. The DMF solution of the isobutyl chloroformate of equimolar quantity is added in the above-mentioned mixed solution under being stirred dropwise, after reacting for 30 minutes, add the DMF solution of the 7-amino-4-methyl-2-quinolinone of equimolar quantity, React for 5 hours under the condition of avoiding light and stirring.

[0037] Preparation of rare earth complexes: equimolar amounts of rare earth Eu 3+ Aqueous solution (Eu 3+ hydrochloride) was mixed with the above conjugate solution for 10 minutes.

[0038] Preparation of rare earth nanoparticles: Mix cyclohexano...

technical field

[0001] The invention relates to a rare earth nano particle emitting light based on the principle of fluorescence energy transfer and a preparation method thereof, which is mainly used for label detection and tracing of biomolecules, and belongs to the technical field of preparation of biomolecular nano-markers. Background technique

[0002] Fluorescent labeling and detection technology has a wide range of applications in life sciences, medicine, immunology and other fields. However, traditional fluorescence analysis is susceptible to interference from nonspecific fluorescence. Most of the currently used fluorescent markers are various organic dye molecules. Since the excitation wavelength is close to the emission wavelength and the Stokes shift is small (20-30nm), the detection is easily interfered by the excitation light. In addition, the background fluorescence and scattered light almost cover the entire range of the fluorescence emission spectrum (350-600...

Examples

Embodiment 1

[0026] Embodiment 1: Preparation of 7-amino-4-methyl-2-quinolinone-DTPA-Tb nanoparticles modified by oligonucleotide probe:

[0027] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: dissolve equimolar amounts of 7-amino-4-methyl-2-quinolinone and diethyltriaminepentaacetic dianhydride in anhydrous In dimethylformamide (DMF), add an appropriate amount of anhydrous triethylamine to the DMF solution of diethyltriaminepentaacetic dianhydride and add 7-amino-4-methyl-2-quinolinone with dropwise stirring DMF solution, reacted for 2 hours under the condition of avoiding light and stirring.

[0028] Preparation of rare earth complexes; equimolar amounts of rare earth Tb 3+ Aqueous solution (Tb 3+ Nitrate or hydrochloride) was mixed with the above-mentioned conjugate solution for 10 minutes.

[0029] Preparation of rare earth nanoparticles: Mix cyclohexanone, n-hexanol, Triton X-100 (v / v, 4:1:1) and stir evenly, add an appropriate amoun...

Embodiment 2

[0031] Embodiment 2: Preparation of aminofluorescein-DTPA-Yb nanoparticles:

[0032] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: Dissolve equimolar amounts of aminofluorescein and diethyltriaminepentaacetic dianhydride in anhydrous dimethylformamide (DMF), Add an appropriate amount of anhydrous triethylamine to the DMF solution of diethyltriaminepentaacetic dianhydride, stir the DMF solution of aminofluorescein dropwise, and react for 2 hours under the condition of avoiding light and stirring.

[0033] Preparation of rare earth complexes: equimolar amounts of rare earth Yb 3+ Aqueous solution (Yb 3+ hydrochloride) was mixed with the above conjugate solution for 10 minutes.

[0034] Preparation of rare earth nanoparticles: Mix cyclohexanone, n-hexanol, Triton X-100 (v / v, 4:1:1) and stir evenly, add an appropriate amount of rare earth complex solution (water phase), and fully stir to form After the water microemulsion, add t...

Embodiment 3

[0035] Example 3: Preparation of 7-amino-4-methyl-2-quinolinone-TTHA-Eu nanoparticles:

[0036] The preparation of the ligand of the rare earth complex and the sensitizing molecule conjugate: dissolve a certain molar amount of triethylenetetraminehexaacetic acid (TTHA) in anhydrous dimethylformamide (DMF), and add a small amount of anhydrous Triethylamine was reacted for 12 hours under stirring conditions. The DMF solution of the isobutyl chloroformate of equimolar quantity is added in the above-mentioned mixed solution under being stirred dropwise, after reacting for 30 minutes, add the DMF solution of the 7-amino-4-methyl-2-quinolinone of equimolar quantity, React for 5 hours under the condition of avoiding light and stirring.

[0037] Preparation of rare earth complexes: equimolar amounts of rare earth Eu 3+ Aqueous solution (Eu 3+ hydrochloride) was mixed with the above conjugate solution for 10 minutes.

[0038] Preparation of rare earth nanoparticles: Mix cyclohexano...

Images