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Preparation method and application of super-resolution imaging probe based on silicon dioxide

A super-resolution imaging, silicon dioxide technology, applied in the analysis of materials, material excitation analysis, material analysis by optical means, etc., can solve the problems of microscopic world imaging that cannot be smaller than 200 nanometers, and achieve excellent scintillation characteristics and high positioning accuracy. , The effect of the simple preparation method

Inactive Publication Date: 2020-04-21
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Due to the limitation of Abbe's diffraction limit, traditional optical microscopes are generally unable to image the microscopic world smaller than 200 nanometers

Method used

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  • Preparation method and application of super-resolution imaging probe based on silicon dioxide
  • Preparation method and application of super-resolution imaging probe based on silicon dioxide
  • Preparation method and application of super-resolution imaging probe based on silicon dioxide

Examples

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Effect test

Embodiment 1

[0041] Dissolve 9.1mg of L-arginine in 6.9mL of deionized water, mix well, add 450 μL of cyclohexane to the solution, heat the reaction bath at 60°C on a heating platform, then add 550 μL of TEOS, 60 temperature water bath and stirred for 20 hours, the resulting solution is the silica nanosphere solution (the total concentration of silica is 20mg / mL).

[0042]Take 1 mL of the silica nanosphere solution and centrifugally purify it into 1 mL of ethanol, and then add 5 μL APTMS to obtain an aminated silica nanosphere solution. Centrifuge and purify into 1 mL of ethanol, then add 2 μL of 1 mg / mL Alexa Fluor647 NHS ester, react in a shaker at room temperature for 12 hours, and purify the sample 6 times at 18000 round / min for 25 min to obtain Alexa Fluor 647 silica nanospheres. Subsequently, 330 μL of the silica nanosphere solution mixed with Alexa Fluor 647 was added to 3.3 μL of 10% (v / v) glutaraldehyde solution, and the reaction was shaken on a shaker at room temperature for 0.5...

Embodiment 2

[0046] Dissolve 9.1 mg of L-arginine in 6.9 mL of deionized water, mix well, add 450 μL of cyclohexane to the solution, heat the reaction bath at 60°C on a heating platform, and then add 550 μL of TEOS, 60 temperature water bath and magnetically stirred for 20 hours, the resulting solution was the silica nanosphere solution (total silica concentration 20 mg / mL).

[0047] 1 mL of the silica nanosphere solution was centrifuged and purified into 1 mL of ethanol, and then 7 μL of APTMS was added to obtain an aminated silica nanosphere solution. Centrifuge and purify into 1 mL of ethanol, then add 3 μL of 1 mg / mL AlexaFluor 647 NHS ester, react on a shaking table at room temperature for 16 hours, and purify the sample 6 times at 18,000 round / min for 30 min to obtain Alexa-incorporated Silica nanospheres of Fluor 647. Subsequently, take 330 μL of the silica nanosphere solution mixed with AlexaFluor 647, add 16.5 μL of 10% (v / v) glutaraldehyde solution, and shake the reaction at roo...

Embodiment 3

[0049] Dissolve 9.1 mg of L-arginine in 6.9 mL of deionized water, mix well, add 450 μL of cyclohexane to the solution, heat the reaction bath at 60°C on a heating platform, and then add 530 μL of TEOS, 60 ℃ constant temperature water bath and stirred for 24 hours, the resulting solution is the silica nanosphere solution (the total concentration of silica is 19 mg / mL).

[0050] 1 mL of the silica nanosphere solution was centrifuged and purified into 1 mL of ethanol, and then 6 μL of APTMS was added to obtain an aminated silica nanosphere solution. Purify by centrifugation into 1 mL of ethanol, then add 2 μL of 1 mg / mL AlexaFluor 647 NHS ester, react in a shaker at room temperature for 12 hours in the dark, and purify the sample 6 times at 18,000 round / min for 30 min to obtain Alexa-incorporated Silica nanospheres of Fluor 647. Subsequently, take 330 μL of the silica nanosphere solution mixed with AlexaFluor 647, add 3.3 μL of 10% (v / v) glutaraldehyde solution, and shake the r...

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Abstract

The invention discloses a super-resolution imaging probe based on silicon dioxide and a preparation method and application of the probe.The super-resolution imaging probe includes silicon dioxide nanoparticles connected with an Alexa Fluor 647 dye, and the surface of the super-resolution imaging probe is coupled with an HER2 (human epidermal growth factor receptor-2) nucleic acid aptamer; the probe can be used for targeted recognition of HER2 overexpression cell exosomes, that is, the HER2 overexpression cell exosomes are subjected to fluorescence labeling; super-resolution optical imaging canbe carried out on the cell exosome which specifically recognizes the probe by utilizing an ultra-high resolution microscope; the probe is uniform in particle size, small in size, high in positioningprecision, excellent in scintillation performance and suitable for ultrahigh-resolution optical imaging based on a single-molecule positioning method; and the exosome is marked by the super-resolutionimaging probe, and the exosome of the cell can be accurately marked by using a single-molecule positioning microscope in a double-channel co-positioning mode.

Description

technical field [0001] The invention relates to the fields of super-resolution imaging probes and exosome labeling, and mainly relates to a silicon dioxide-based super-resolution imaging probe and its preparation method and application. Background technique [0002] Due to the limitation of Abbe's diffraction limit, traditional optical microscopes are generally unable to image the microscopic world smaller than 200 nanometers. In recent decades, under the unremitting exploration of scientific researchers, super-resolution imaging technology has emerged as the times require. For the first time, we humans can observe ultra-high-resolution images within the diffraction limit. [0003] Single-molecule localization-based super-resolution fluorescence microscopy (SMLM) includes photoactivated localization imaging (PALM) and stochastic optical reconstruction super-resolution imaging (STORM), which combines organic fluorescent probes and super-resolution optical microscopy technique...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6458G01N2021/6439
Inventor 宗慎飞袁江南王著元崔一平
Owner SOUTHEAST UNIV
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