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Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof

An optical imaging, dual-targeting technology, applied in the field of SERS-fluorescence dual-mode optical imaging probes, can solve problems such as the limitations of fluorescence multiplex imaging, and achieve the effect of enhancing target recognition and efficiency, high sensitivity, and improving endocytosis

Inactive Publication Date: 2016-08-10
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although fluorescence imaging is fast and intuitive, fluorescence multiplex imaging is severely limited due to its wide spectral bandwidth

Method used

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  • Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof
  • Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof
  • Dual-mode optical imaging probe with tumor double-targeting function and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] To prepare a dual-mode probe with enhanced targeting effect on cervical cancer cells (HeLa), the preparation method is as follows:

[0046] 1) The experiment uses the seed growth method to prepare a uniform spherical silver nanoparticle solution. Under vigorous stirring, 5 mL of 1% sodium citrate aqueous solution was added to boiling 245 mL of silver nitrate solution (containing 90 mg of silver nitrate), and stirring was continued for one hour to obtain a silver glue solution. Take 10mL of silver gel solution and disperse it in alcohol solution by centrifugation, add 10uL of 10mM Raman reporter 4MBA, mix and stir for 1-4 hours, observe whether the color of the solution changes to prevent aggregation;

[0047] 2) Add 200μL of ammonia water and 5μL of tetraethoxysilane solution to the solution of step 1, and stir at room temperature for more than 4 hours, and observe that the solution turns into khaki;

[0048] 3) Add 200μL of ammonia, 5μL of 1mg / mL 3-aminopropyltriethoxysilane...

Embodiment 2

[0053] Furthermore, the dual-targeting dual-mode fluorescent probe is used in dual-mode imaging of cervical cancer cells HeLa, the specific method is as follows:

[0054] 1) Inoculate cervical cancer cells in a petri dish, add MEM cell culture medium containing 10% fetal bovine serum and 1% penicillin-streptomycin mixture, and place it in an incubator at 37°C and 5% CO2 for 24 hour. After that, add the probe at a 4:1 ratio of culture solution to probe, mix well and place it in the incubator for two hours. After the particles are internalized by the cells, the culture medium is removed, the cells are washed 3 times with PBS, fixed with 4% paraformaldehyde for 10 minutes, and washed twice with PBS for later use.

[0055] 2) Add a small amount of buffer to the fixed cells obtained in step 1, adjust the focal plane to focus, select a cell area, excite with 543nm laser, 560nm-600nm as the receiving wavelength range, imaging to obtain cell fluorescence images, such as Figure 5 Shown. ...

Embodiment 3

[0058] Furthermore, in order to show that the targeting ability of the dual-targeting probe is better than that of the traditional single-targeting probe, the following experiments were performed to verify its targeting ability:

[0059] 1) Prepare a single-targeting probe according to the steps in Example 1. The only difference is that in the last step, only one polypeptide of the same concentration as the double-targeting probe is added to the solution. Here, cys-HAIYPRH is selected as an example.

[0060] 2) Inoculate cervical cancer cells in two culture dishes at the same concentration, add MEM cell culture medium containing 10% fetal bovine serum and 1% penicillin-streptomycin mixture, and place in an incubator at 37°C, 5% CO 2 Cultivate for 24 hours in the environment. After that, the two probes were added with a 4:1 ratio of culture solution to probe, mixed evenly and then placed in the incubator for two hours. After the particles are internalized by the cells, the culture m...

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Abstract

The invention discloses preparation and application of an SERS-fluorescence dual-mode optical imaging probe with a tumor double-targeting function. The nano-probe is composed of an imaging function core part and a targeting function shell part. The imaging function core part comprises silver nanoparticles adsorbed with Raman reporter molecules and a silicon dioxide shell layer doped with fluorescence dye, so that the probe has the SERS imaging function and the fluorescence imaging function simultaneously. The targeting function shell comprises a polyethylene glycol (PEG) coupling agent and two polypeptides specifically targeting tumor cells. According to the probe, the more effective targeting capacity is supplied to the nanoparticles by adopting a dual-polypeptide modification method, therefore, the probe can well enter the tumor cells, recognition of the probe to the tumor cells is enhanced, and SERS-fluorescence dual-mode imaging on a tumor is achieved.

Description

Technical field [0001] The invention belongs to the technical field of bioanalytical chemistry, and specifically relates to a tumor dual-targeted SERS-fluorescence dual-mode optical imaging probe and its preparation and application. Background technique [0002] Nanotechnology has a broad application prospect in tumor diagnosis and treatment. Nano drug carriers and imaging contrast agents that have the function of directional recognition and killing tumor cells have always been a major direction of nanotechnology research. How to better enrich the nanoparticles in tumor cells as much as possible, thereby reducing the damage to normal cells, is the goal of scientists' unremitting efforts. Nanoparticle targeting is divided into passive targeting and active targeting. Passive targeting refers to the use of the EPR effect of solid tumor blood vessels to allow nanocarriers smaller than 200nm to leak from the gaps in the tumor blood vessels and passively accumulate and stay in the tum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0002A61K49/0013A61K49/0056A61K49/0093
Inventor 王著元张益之伍磊宗慎飞崔一平
Owner SOUTHEAST UNIV
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