Raman silence region substrate with gold as core and dopamine as shell and preparation method and application of substrate

A technology of Raman silent zone and nuclear dopamine, applied in the Raman silent zone "probe molecule and its preparation, and the application field of intracellular Raman imaging, achieving great clinical application prospects and low-cost effects

Active Publication Date: 2016-12-14
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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This material enables highly sensitive ...

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  • Raman silence region substrate with gold as core and dopamine as shell and preparation method and application of substrate
  • Raman silence region substrate with gold as core and dopamine as shell and preparation method and application of substrate
  • Raman silence region substrate with gold as core and dopamine as shell and preparation method and application of substrate

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Embodiment 1

[0023] Example 1: Synthesis of a surface-enhanced Raman scattering substrate for intracellular imaging of human cervical cancer cells with gold as the core and dopamine reducing silver nitrate as the shell

[0024] Take 1-3 mL of perchlorauric acid solution with a concentration of 8-10 mg / mL and 90-100 mL of deionized water into a 250 mL three-neck flask, mix well, and heat in an oil bath at 120-140 °C , control it to reflux about once per second, at this time, add 600-800 uL trisodium citrate with a concentration of 1-3% to it, keep reflux for 10-30min, cool to room temperature, and obtain gold nanoparticles with uniform size. Store it in a clean glass bottle at 4-8°C;

[0025]Take the synthesized gold nanoparticles, centrifuge them at 8000 rpm for 5 min, absorb the supernatant, redissolve it in borax solution, add 10 uL of 10 mM (E)-2-((4-(acetylene Base) benzylidene) amino) ethanethiol, mix well, add 5 uL of 10 mM silver nitrate, and finally add 60 uL of 1 mg / mL dopamine. ...

Embodiment 2

[0026] Example 2: Application of a Raman probe with gold as the nucleus and dopamine reducing silver nitrate as the shell to enhanced Raman imaging of the inner surface of glioma cell U251

[0027] First add about 10 5 After culturing for 12-24 hours, take 10 μL of surface-enhanced Raman scattering material of gold as the core dopamine-reduced silver nitrate as the silent area of ​​the shell, add it to a glass dish, and then put the glass culture dish in the cell culture React with the cells in the box for 12-24 hours, then absorb the culture medium, wash with PBS for two to three times, react with 4% paraformaldehyde solution in a 37°C incubator for 15-30 minutes, and finally wash with PBS for two to three times Three times, and add 100-200uL of PBS and store in a refrigerator at 4°C for surface-enhanced Raman scattering imaging.

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Abstract

The invention belongs to the technical field of biomacromolecule separation and analysis, and particularly relates to a Raman silence region substrate with gold as a core and dopamine as a shell and a preparation method and application of the substrate. The preparation method includes the steps that gold nanoparticles are directly reduced through the hydrothermal method and then dissolved in a borax solution after being centrifuged, and Raman silence region molecules (E)-2-((4-(acetenyl)benzal)amino)ethanethio prepared through the hydrothermal method are added; then a trace amount of silver nitrate is added, then dopamine is added, and accordingly silver nanoparticles are reduced in situ; by means of auto-polymerization of dopamine, the stable Raman substrate of the Raman silence region is generated. Experiments represent that novel Rama probe molecules achieve an excellent Raman imaging effect on the insides of cells under the enhancement function of enhancement surfaces of the gold nanoparticles. The substrate is free of template and low in cost and toxicity, is applied to live cell imaging, and is easy to operate, novel, convenient to use, practical and efficient.

Description

technical field [0001] The invention belongs to the technical field of separation and analysis of biological macromolecules, and in particular relates to a "Raman silent region" probe molecule, a preparation method thereof and an application in intracellular Raman imaging. Background technique [0002] Surface-enhanced Raman scattering is on the rough surface of some precious metals such as gold, silver, copper, etc., and the Raman scattering signal of the molecule to be measured is affected by the surface plasmon resonance (LSPR) by a geometric multiple (10 3 -10 9 )enlarge. At the same time, due to the high sensitivity and good spectral shape of Raman spectroscopy, it is widely used in various fields of high-sensitivity analysis and research. In addition, because the Raman signal can be stabilized in a variety of chemical environments, and the structural information of the measured molecule can be displayed in the spectrum analysis, it provides strong technical support f...

Claims

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Application Information

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IPC IPC(8): G01N21/65
Inventor 高明霞陈萌张祥民
Owner FUDAN UNIV
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