A kind of core-shell nano-silica fluorescent probe and its synthesis method and application

A nano-silica, fluorescent probe technology, applied in nano-optics, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of difficult microbial functional activity tracking and other problems, achieve high-sensitivity analysis and sorting, major practice The effect of meaning and application potential

Active Publication Date: 2019-11-22
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the fluorescent probes used in strain screening studies in the past were general-purpose nucleic acid probes, which made it difficult to track the functional activities of microorganisms.

Method used

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  • A kind of core-shell nano-silica fluorescent probe and its synthesis method and application
  • A kind of core-shell nano-silica fluorescent probe and its synthesis method and application
  • A kind of core-shell nano-silica fluorescent probe and its synthesis method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The schematic diagram of the synthesis process of dye Rod-X and dye Methyl-S is as follows: Figure 14 As shown, the synthesis flow chart of NP-1 and NP-2 is as follows Figure 15 As shown, the specific synthesis process is as follows.

[0036] 1) Synthesis of dye Rod-X:

[0037] Add 0.48g rhodamine B, 1.46g allyl bromide (12mmol), 2.65g Cs to a 100mL volumetric flask 2 CO 3 Dissolve in 50 mL of dry DMF, heat to 80°C under nitrogen protection, react overnight, add 200 mL of dichloromethane, wash the organic phase with brine three times, dry the organic phase with anhydrous sodium sulfate, remove the organic solvent by distillation under reduced pressure, pass through silica gel column separation to obtain product 0.31g, yield 64% (methanol / dichloromethane=1:40, R f = 0.2). 1 H NMR (400MHz, CDCl 3 ) δ1.3(t,CH 3 ,12H),3.5(q,CH 2 ,8H),4.5(d,CH 2 ,2H),5.1(t,CH 2 ,2H),5.7(qd,CH,1H),6.8(s,Ar,2H),6.9(d,Ar,2H),7.0(d,Ar,2H),7.3(m,Ar,1H),7.8 (td,Ar,2H),8.32(d,Ar,1H).HR...

Embodiment 2

[0047] Spectra of the nanoprobe:

[0048] Dye Rod-X and dye Methyl-S were dissolved in ethanol, nanoparticles were dispersed in aqueous solution, and their infrared, absorption and fluorescence spectra were tested. The infrared spectrum test uses a Fourier transform infrared spectrometer (Bruker / TensorII), with a test range of 4000-400cm -1 . The ultraviolet spectrum test uses an ultraviolet-visible spectrometer (SHIMADZU / UV-2600), and the test range is 400-800nm. The fluorescence spectrum test uses a fluorescence spectrophotometer (Perkin Elmer / LS-45), the excitation wavelength range is 200-600nm, and the emission wavelength range is 400-800nm. The luminescence of the nanoparticles was observed using a hand-held UV lamp.

[0049] figure 1 It is the spectral analysis of the nanoprobe and the dye of the present invention. A is the absorption spectrum and fluorescence emission spectrum of the dye Rod-X, the maximum absorption wavelength is 555nm, the maximum emission wavele...

Embodiment 3

[0051] The morphology and structure of nanoparticles:

[0052] Dry the nanoparticles at various stages in a vacuum drying oven at 60°C overnight, grind them into powder after fully drying, and perform thermogravimetric analysis test. The test temperature range is 30-700°C, and the heating rate is 10°C / min. Next test. Disperse the nano fluorescent probe in the aqueous solution, observe the shape and size of the particle by transmission electron microscopy (TEM), test its particle size distribution (DLS) by dynamic light scattering ( Figure 2-4 ).

[0053] figure 2 is the thermogravimetric analysis of nanoparticles in various stages of the present invention. Panel A is the thermogravimetric spectrum of silica nanoparticles without dye binding, which has two weight loss stages and loses about 9% of its weight. B is the thermogravimetric spectrum of NP-1, which has two weight loss stages and loses about 10% of its weight. C is the thermogravimetric spectrum of NP-2, which h...

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Abstract

The invention discloses a core-shell nano-silica fluorescent probe, a synthesis method and an application thereof. The core-shell nano-silica fluorescent probe of the present invention is a nanoparticle of a core-shell structure, and the core-shell structure includes an inner fluorescent dye shown in formula (I) and a shell dye shown in formula (II), and the nanoparticle The structure is shown in formula (III); the present invention also discloses a method for using a core-shell nano-silica fluorescent probe to select and breed microorganisms with toxic aromatic hydrocarbon degradation activity. The technology-based microbial breeding research mode can perform rapid and highly sensitive analysis and sorting of single cells more efficiently, intuitively and specifically, and can provide new tools for efficient breeding of functional microorganisms, and can also provide a basis for single-cell research and development of functional microorganisms. The in-depth excavation of uncultivated microorganisms provides technical support, which has great practical significance and application potential for the prevention and control of toxic aromatic hydrocarbon pollutants.

Description

technical field [0001] The invention belongs to the technical field of functional microorganism breeding. More specifically, the invention relates to a core-shell nano-silica fluorescent probe, its synthesis method and its application in functional microorganism breeding. Background technique [0002] With the rapid development of modern industry, the output and types of synthetic aromatic compounds are increasing day by day. These compounds have complex structures and are difficult to degrade naturally. They have biological toxicity such as carcinogenicity and teratogenicity to humans, and can migrate and diffuse long distances with media such as air, water, soil, and organisms, causing extensive non-point source pollution and seriously threatening human beings. health and ecological safety. Therefore, the pollution control of aromatic hydrocarbons is a major problem in ecological environment protection. [0003] Microorganisms are important drivers of material cycle and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D311/02C09K11/02C09K11/06B82Y20/00B82Y30/00B82Y40/00G01N21/64
CPCG01N21/6428C09K11/02C09K11/06B82Y20/00B82Y30/00B82Y40/00C12Q1/04G01N33/588C09B67/0033C09B69/008C09B69/103C09B69/106C09B67/0097C09B11/24C09B29/08G01N33/56911G01N2021/6439
Inventor 许玫英刘飞廖兵罗业燊李恩泽
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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