Gold nano-star @ quantum dot composite cell probe with photothermal and fluorescence enhancement dual-functions and preparation method and applications thereof

Abstract

The invention provides a gold nano-star @ quantum dot composite cell probe with photothermal and fluorescence enhancement dual-functions, which sequentially comprises a gold nano-star, a silica layer, quantum dots and a silica nano-shell layer from inside to outside. The cell probe is prepared by using a chemical layer-by-layer growth method implemented through the following steps of: firstly, preparing a gold nano-star by using a seed crystal growth method; coating a silica layer on the outside of the gold nano-star by means of hydrolysis so as to form a silica sphere coated with the gold nano-star; then, linking quantum dots on the surface of the silica sphere through a crosslinking reaction so as to form a silica sphere composite material internally coated with the gold nano-star and externally linked with the quantum dots; and finally, coating a silica nano-shell layer on the surface of the composite material. Compared with traditional probes, the probe integrates the functions of photothermal treatment and fluorescence labeling; the fluorescence intensity of the quantum dots is enhanced, and the biological toxicities of the gold nano-star and the quantum dots are effectively avoided; and in addition, the cell probe has extremely good biological compatibility.

Description

technical field

[0001] The invention relates to the technical field of composite nanoprobes, in particular to a gold nanostar quantum dot composite cell probe with dual functions of photothermal and fluorescence enhancement and its preparation method and application. Background technique

[0002] Photothermal material is a kind of light that can absorb certain light, especially near-infrared light (because its wavelength range endows it with unique optical safety properties, it can penetrate human skin and deep tissue, and is the skylight of the lesion). The heat generated by plasmon resonance or energy transition bands can cause high temperature locally, and finally kill the nano-functional materials of tumor cells. Photothermal conversion functional materials are favored because they can convert near-infrared light into high heat, and have become a research hotspot in biological applications. Various gold nanostructures (gold nanorods, gold nanoshells, gold nanocages, etc...

Examples

preparation example Construction

[0041] (1) Preparation of gold nanostars:

[0042] Dissolve 1-10ml of 1mM HAuCl 4 After the solution is boiled, add 0.1-5ml of 1% sodium citrate (or sodium borohydride, ascorbic acid, tetrakishydroxymethyl phosphorus chloride) solution to it, and heat (at a temperature of 25-100 degrees) to react 5- 400 minutes to obtain the size of the 5-200nm gold seed crystal solution; to 5-100mL of 1mM HAuCl 4 Add 10-5000uL of the above gold seed crystal solution (mixed with 50uL HCl solution) and stir for 5 minutes, then add 20-4000uL of 2M AgNO 3 The solution was stirred for 30s, and then centrifuged for later use; the size of the prepared gold nanostars was adjustable from 20-400nm;

[0043] (2) Preparation of gold nanostar quantum dot composites:

[0044] Dissolve 4 mL of prepared gold nanostars in a mixed solution of ethanol and water (preferably, the amount of the mixed solution of ethanol and water is based on the ability to dissolve gold nanostars, wherein the ethanol: the volume ...

Embodiment 1

[0049] (1) Boil 4mL of 1mM HAuCl4 solution, add 0.7mL of 1% sodium citrate solution to it, heat at 80°C, and react for 15min to obtain a gold seed crystal solution. To 10 mL of 1 mM HAuCl 4 (mixed with 50uL HCl solution), add 100uL gold seed crystal solution and stir for 5 minutes, then add 100uL 2M AgNO 3 The solution was stirred for 30 s, and then centrifuged for later use.

[0050] (2) Dissolve 4 mL of the prepared gold nanostars in a mixture of ethanol and water (preferably, the amount of the mixture of ethanol and water is subject to the ability to dissolve gold nanostars, wherein the ethanol: water The volume ratio is preferably 4:1), add 2mL of ammonia water, and then drop 300uL of tetraethyl orthosilicate (TEOS) into the reaction solution to coat the gold nanostars with silica spherical shells. After 24 hours of reaction Perform centrifugal washing; then 300uL of 3-aminopropyltrimethoxysiloxane (APTMS) was used to modify the surface of the silicon sphere, and then 3m...

Embodiment 2

[0053] (1) Add 4mL of 1mM HAuCl 4 After the solution was boiled, 1.0mL of 1% sodium citrate solution was added to it, and the gold seed crystal solution was obtained after reacting at 60°C for 15 minutes. To 10 mL of 1 mM HAuCl 4 (mixed with 50uL HCl solution), add 200uL gold seed crystal solution and stir for 5 minutes, then add 100uL 2M AgNO 3 The solution was stirred for 30 s, and then centrifuged for later use.

[0054] (2) Dissolve 4 mL of the prepared gold nanostars in a mixture of ethanol and water (preferably, the amount of the mixture of ethanol and water is subject to the ability to dissolve gold nanostars, wherein the ethanol: water The volume ratio is preferably 4:1), add 2mL of ammonia water, and then add 600uL of tetraethyl orthosilicate (TEOS) dropwise into the reaction solution to coat the gold nanostars with silica spherical shells. After 24 hours of reaction Perform centrifugal washing; then 600uL of 3-aminopropyltrimethoxysiloxane (APTMS) was used to modi...

technical field

[0001] The invention relates to the technical field of composite nanoprobes, in particular to a gold nanostar quantum dot composite cell probe with dual functions of photothermal and fluorescence enhancement and its preparation method and application. Background technique

[0002] Photothermal material is a kind of light that can absorb certain light, especially near-infrared light (because its wavelength range endows it with unique optical safety properties, it can penetrate human skin and deep tissue, and is the skylight of the lesion). The heat generated by plasmon resonance or energy transition bands can cause high temperature locally, and finally kill the nano-functional materials of tumor cells. Photothermal conversion functional materials are favored because they can convert near-infrared light into high heat, and have become a research hotspot in biological applications. Various gold nanostructures (gold nanorods, gold nanoshells, gold nanocages, etc...

Examples

preparation example Construction

[0041] (1) Preparation of gold nanostars:

[0042] Dissolve 1-10ml of 1mM HAuCl 4 After the solution is boiled, add 0.1-5ml of 1% sodium citrate (or sodium borohydride, ascorbic acid, tetrakishydroxymethyl phosphorus chloride) solution to it, and heat (at a temperature of 25-100 degrees) to react 5- 400 minutes to obtain the size of the 5-200nm gold seed crystal solution; to 5-100mL of 1mM HAuCl 4 Add 10-5000uL of the above gold seed crystal solution (mixed with 50uL HCl solution) and stir for 5 minutes, then add 20-4000uL of 2M AgNO 3 The solution was stirred for 30s, and then centrifuged for later use; the size of the prepared gold nanostars was adjustable from 20-400nm;

[0043] (2) Preparation of gold nanostar quantum dot composites:

[0044] Dissolve 4 mL of prepared gold nanostars in a mixed solution of ethanol and water (preferably, the amount of the mixed solution of ethanol and water is based on the ability to dissolve gold nanostars, wherein the ethanol: the volume ...

Embodiment 1

[0049] (1) Boil 4mL of 1mM HAuCl4 solution, add 0.7mL of 1% sodium citrate solution to it, heat at 80°C, and react for 15min to obtain a gold seed crystal solution. To 10 mL of 1 mM HAuCl 4 (mixed with 50uL HCl solution), add 100uL gold seed crystal solution and stir for 5 minutes, then add 100uL 2M AgNO 3 The solution was stirred for 30 s, and then centrifuged for later use.

[0050] (2) Dissolve 4 mL of the prepared gold nanostars in a mixture of ethanol and water (preferably, the amount of the mixture of ethanol and water is subject to the ability to dissolve gold nanostars, wherein the ethanol: water The volume ratio is preferably 4:1), add 2mL of ammonia water, and then drop 300uL of tetraethyl orthosilicate (TEOS) into the reaction solution to coat the gold nanostars with silica spherical shells. After 24 hours of reaction Perform centrifugal washing; then 300uL of 3-aminopropyltrimethoxysiloxane (APTMS) was used to modify the surface of the silicon sphere, and then 3m...

Embodiment 2

[0053] (1) Add 4mL of 1mM HAuCl 4 After the solution was boiled, 1.0mL of 1% sodium citrate solution was added to it, and the gold seed crystal solution was obtained after reacting at 60°C for 15 minutes. To 10 mL of 1 mM HAuCl 4 (mixed with 50uL HCl solution), add 200uL gold seed crystal solution and stir for 5 minutes, then add 100uL 2M AgNO 3 The solution was stirred for 30 s, and then centrifuged for later use.

[0054] (2) Dissolve 4 mL of the prepared gold nanostars in a mixture of ethanol and water (preferably, the amount of the mixture of ethanol and water is subject to the ability to dissolve gold nanostars, wherein the ethanol: water The volume ratio is preferably 4:1), add 2mL of ammonia water, and then add 600uL of tetraethyl orthosilicate (TEOS) dropwise into the reaction solution to coat the gold nanostars with silica spherical shells. After 24 hours of reaction Perform centrifugal washing; then 600uL of 3-aminopropyltrimethoxysiloxane (APTMS) was used to modi...

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