Aggregation-induced luminescent nanometer fluorescent probe and preparation method thereof

Abstract

The present invention discloses an aggregation induced luminescence nanofluorescent probe which coats a chitosan fluorescent molecule with the characteristic of aggregation induced luminescence on the surface of a hydroxyl apatite nanoparticle, and the structural formula of the chitosan fluorescent molecule is shown in a formula (1). The aggregation induced luminescence nanofluorescent probe is prepared by a chemical co-precipitation process. The size of the aggregation induced luminescence nanofluorescent probe is about 110 nm, the dispersion is stable, positive changes are formed on the surface thereof, and the aggregation induced luminescence nanofluorescent probe has the feature of the aggregation induced luminescence. Compared with a traditional fluorescent probe, the aggregation induced luminescence nanofluorescent probe has the advantages that the sensitivity is high, the light stability is good, and quenching does not exist during high concentration and a fluorescence spectrum is not drifted, so that the aggregation induced luminescence nanofluorescent probe can be applied to the fields of bioimaging, fluorescent tracing detection and the like. The aggregation induced luminescence nanofluorescent probe is shown in the figure below.

Description

technical field

[0001] The invention relates to a method for preparing a fluorescent probe, in particular to a method for preparing a hydroxyapatite / chitosan nanometer fluorescent probe with aggregation-induced luminescent properties. Background technique

[0002] A probe is a molecule that can interact with a specific target molecule to detect the target molecule, and requires no or only negligible interference to the detected object after the interaction. Fluorescent probes use fluorescent substances as indicators, and under the excitation of a certain wavelength of light, the indicator produces fluorescence, and the qualitative or quantitative analysis of the detected substance is realized by detecting the generated fluorescence. In the past two decades, the use of fluorescent probes to detect biological processes in living cells or animals has attracted widespread attention from chemists and biologists, and a variety of fluorescent biological probes have been developed, ...

Examples

Embodiment 1

[0022] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.0294 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0023] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0024] 3) AIE fluorescent-labeled chitosan with a labeling rate of 0.1 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 1 mg / mL;

[0025] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepar...

Embodiment 2

[0028] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.0557 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0029] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0030] 3) AIE fluorescent-labeled chitosan with a labeling rate of 20 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 3 mg / mL;

[0031] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepare...

Embodiment 3

[0034] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.147 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0035] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0036] 3) AIE fluorescent-labeled chitosan with a labeling rate of 10 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 5 mg / mL;

[0037] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepared...

technical field

[0001] The invention relates to a method for preparing a fluorescent probe, in particular to a method for preparing a hydroxyapatite / chitosan nanometer fluorescent probe with aggregation-induced luminescent properties. Background technique

[0002] A probe is a molecule that can interact with a specific target molecule to detect the target molecule, and requires no or only negligible interference to the detected object after the interaction. Fluorescent probes use fluorescent substances as indicators, and under the excitation of a certain wavelength of light, the indicator produces fluorescence, and the qualitative or quantitative analysis of the detected substance is realized by detecting the generated fluorescence. In the past two decades, the use of fluorescent probes to detect biological processes in living cells or animals has attracted widespread attention from chemists and biologists, and a variety of fluorescent biological probes have been developed, ...

Examples

Embodiment 1

[0022] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.0294 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0023] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0024] 3) AIE fluorescent-labeled chitosan with a labeling rate of 0.1 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 1 mg / mL;

[0025] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepar...

Embodiment 2

[0028] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.0557 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0029] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0030] 3) AIE fluorescent-labeled chitosan with a labeling rate of 20 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 3 mg / mL;

[0031] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepare...

Embodiment 3

[0034] 1) Weigh 0.0555 g of anhydrous calcium chloride, 0.0468 g of sodium dihydrogen phosphate and 0.147 g of trisodium citrate hydrate into a 20 mL strain bottle, add 10 mL of three-distilled water, and add 200 μL of glacial acetic acid dropwise under magnetic stirring Dissolved to make 5 mg / mL hydroxyapatite precursor solution;

[0035] 2) Dilute the hydroxyapatite precursor solution in step 1) to 1 mg / mL with triple distilled water, and add 40 wt% sodium hydroxide aqueous solution dropwise under stirring to adjust the pH of the solution to 10. At this time, the hydroxyapatite nanoparticles Precipitate, continue to stir overnight, make hydroxyapatite nano particle solution;

[0036] 3) AIE fluorescent-labeled chitosan with a labeling rate of 10 mol% was dissolved in 2% acetic acid aqueous solution to prepare an AIE fluorescent-labeled chitosan-acetic acid solution with a concentration of 5 mg / mL;

[0037] 4) Measure 1 mL of the hydroxyapatite nanoparticle solution prepared...

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