CN111879753A - Noble metal nanocone SERS substrate for detecting novel coronavirus and preparation method thereof - Google Patents
Noble metal nanocone SERS substrate for detecting novel coronavirus and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of laser Raman spectroscopy and virus detection, and particularly relates to a noble metal nanocone SERS substrate for detecting a novel coronavirus and a preparation method thereof. The SERS substrate comprises a substrate, a nanocone array positioned on the substrate and a precious metal film coated on the surface of the substrate where the nanocone and the nanocone are positioned, wherein the nanocones forming the nanocone array are radial from the center of the substrate to the periphery, and included angles between adjacent nanocones are in a gradual change mode, so that effective capture can be realized for coronavirus with the size of 60-140nm, the compatibility is higher, and when the virus is in contact with the substrate and slides on the surface, the capture efficiency can be greatly improved. The preparation method comprises the following steps: the method is simple, the prepared precious metal nanocone has SERS hot spots distributed in three dimensions, and the SERS sensitivity is high.
Description
Technical Field
The invention relates to the technical field of laser Raman spectroscopy and virus detection, in particular to a noble metal nanocone SERS substrate for detecting novel coronavirus and a preparation method thereof.
Background
2019 the novel coronavirus (2019-nCOV) can be transmitted by respiratory droplets, contact, aerosol and the like, symptoms of human infection are mainly fever, cough, shortness of breath and dyspnea, and severe patients can cause renal failure and even death. Because of the lack of specific drugs, the physical isolation of suspected and confirmed cases is an effective way to prevent further worsening of epidemic situations. Therefore, it is very important to rapidly screen patients infected with the new pneumonia from the population. At present, the commonly used screening and detection technology is mainly a reverse transcription-polymerase chain reaction (RT-PCR) method, which not only takes a long time for a diagnosis process (about 3 hours for single detection), but also has a high false negative rate due to poor sensitivity of a detection kit. Therefore, in order to effectively block the spread of the novel coronavirus, a novel rapid field detection technology needs to be developed.
The detection technology based on the Surface Enhanced Raman Scattering (SERS) effect has the advantages of simple and convenient operation, short detection time, low detection limit, fingerprint identification and the like, and has the ultrasensitive detection capability of single molecules. Meanwhile, with the gradual maturity of the portable laser raman spectroscopy technology, SERS has been developed into a rapid, trace, lossless and in-situ field detection technology. At present, a great deal of research has shown that SERS can analyze the base sequence and composition of nucleic acid and the interaction between proteins or ligands, and can identify and detect different RNA or DNA virus cells with high performance, with DNA, RNA, proteins and other biomolecules as detection targets. For example, the wangangglow (Talanta,205,2019) of the university of mail and telecommunications in Nanjing has obtained silver nanorod arrays on glass substrates by an oblique angle deposition method, enabling SERS detection of H7 and N9 subtype influenza viruses. However, in order to Capture the virus, auxiliary reagents including a place chain, a Capture chain and a Probe chain need to be designed based on the virus gene sequence, and a detection sample needs to be co-cultured with the auxiliary reagents at a certain temperature, so the steps of the Capture method of the "bio-bonding type" are complicated and time-consuming. Meanwhile, the traditional precious metal conical SERS substrate has a certain nano-cone distance and a certain nano-cone gap, only viruses with small size distribution can be captured, the capturing efficiency is low, and the characteristic Raman signal intensity required by detection is weak.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a noble metal nanocone SERS substrate for detecting novel coronavirus.
In order to overcome the defects in the prior art, the invention provides a preparation method of the precious metal nanocone SERS substrate for detecting the novel coronavirus.
In order to solve the technical problem, the adopted technical scheme is that the precious metal nanocone SERS substrate for detecting the novel coronavirus comprises a substrate, a nanocone array positioned on the substrate and a precious metal film coated on the surface of the substrate where the nanocone array and the nanocone array are positioned, wherein the thickness of the precious metal film is 10-50nm, and the precious metal film is made of precious metal with Raman activity; the nano cone array comprises a plurality of nano cone rings with different diameters, the nano cone rings are formed by circumferentially arranging a plurality of nano cones around a nano cone perpendicular to a substrate, an included angle alpha formed between the central axis of each nano cone and the substrate is 30-90 degrees, the included angle alpha is gradually reduced from an inner ring of the nano cone ring to an outer ring of the nano cone ring, the height of each nano cone is 200-300nm, the diameter of the root of each nano cone is 50-300nm, the diameter of the tip of each cone is 20-50nm, the tip angle of each cone is 10-60 degrees, and the distance between every two adjacent nano cones is 80-150 nm.
As a further technical scheme of the precious metal nanocone SERS substrate for detecting the novel coronavirus:
preferably, the substrate and the nanocones are made of the same material and are made of flexible materials capable of bending.
As a further preferable material of the substrate and the nanocone, the material of the substrate and the nanocone is one of polyimide and polyethylene terephthalate.
Preferably, the noble metal thin film is formed by stacking noble metal nano-particles with the size of 10-30 nm.
Preferably, the material of the noble metal thin film is one of gold, silver and copper.
Preferably, in the nano-cone array, the distances between adjacent nano-cones are not completely equal or not equal.
In order to solve another technical problem of the present invention, the technical scheme is that a preparation method of a noble metal nanocone SERS substrate for detecting a novel coronavirus includes the following steps:
s1, covering a monolayer of polystyrene microspheres on the surface of the substrate by a self-assembly method, wherein the diameter of the polystyrene microspheres is 80-150 nm;
s2, smoothly bending the periphery of the substrate towards the center of the surface where the polystyrene microspheres are located by 5-30 degrees, and then placing the bent substrate in a reactive ion etching device;
s3, using sulfur hexafluoride as a reaction gas, and performing reactive ion etching on the substrate surface where the polystyrene microspheres are located under the conditions that the gas flow rate is 10-50sccm, the cavity pressure is 5-20 Pa and the etching power is 100-;
s4, keeping the bending state of the substrate, depositing noble metal nanoparticles on the surface of the substrate where the nanocones are located through a magnetron sputtering method to form a noble metal film, and then flatting the bent substrate to obtain the noble metal nanocone SERS substrate for detecting the novel coronavirus.
The preparation method of the noble metal nanocone SERS substrate for detecting the novel coronavirus adopts the further technical scheme that:
preferably, the diameters of the polystyrene microspheres in step S1 are not completely the same or completely different.
Compared with the prior art, the invention has the beneficial effects that:
1) the invention discloses a precious metal nanocone SERS substrate for detecting novel coronavirus, wherein the surface of an SERS detection chip is provided with rough conical protrusions, the nanocones are gradually radially directed from the middle to the periphery, and an included angle formed between the central axis of each nanocone and a substrate is gradually enlarged. When being used for SERS technique to detect novel coronavirus, compare ordinary fixed contained angle's nanometer awl, its advantage that is showing has two points: firstly, the sizes of gaps and angles between the substrate nanocones have the characteristic of gradual change, and the substrate can effectively capture novel coronavirus with the size of 60-140nm, so that the substrate has stronger compatibility; secondly, the orientation of the nanocones (i.e., the opening direction of the cone gap) is radial from the center of the substrate to the periphery, and when the virus is in contact with the substrate and slides on the surface, the capture efficiency can be greatly improved. The efficient and firm capture of the virus on the surface of the substrate is the key for obtaining stable Raman detection signals. Particularly, when virus detection is carried out in an expiratory mode, false negative caused by capture of too few viruses can be effectively avoided, and compared with a particle membrane or a conventional nano-cone array, the method has higher capture efficiency and detection accuracy on the novel coronavirus.
2) The method discloses a preparation method of a noble metal nanocone SERS substrate for detecting novel coronavirus, which comprises the steps of covering a single-layer polystyrene microsphere template on the substrate, then bending the substrate towards the center of the surface of the polystyrene microsphere template, then carrying out reactive ion etching on the surface of the polystyrene microsphere in the substrate, wherein ionized fluorine free radicals can simultaneously carry out reactive etching on the substrate and the microsphere template, because the microspheres play a role in protecting the mask, after the etching is finished, the microspheres on the surface layer are completely removed, the substrate at the bottom is a conical protrusion, noble metal nano particles are deposited on the surface of the substrate where the conical protrusion is located by a magnetron sputtering method to form a noble metal film, then the bent substrate is laid flat, thus obtaining the noble metal nanocone SERS substrate with nanocones with different opening angles and directions. The preparation method is simple, the prepared noble metal nanocone can be used as an active substrate for surface enhanced Raman scattering, and the prepared noble metal nanocone has three-dimensionally distributed SERS hot spots and high SERS sensitivity.
Drawings
FIG. 1 is a flow chart of the preparation of the noble metal nanocone SERS substrate of the present invention;
fig. 2 is a raman spectrum characterization graph of the noble metal nanocone SERS substrate prepared in comparative example 1 and example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments, and all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the protection scope of the present invention.
Comparative example 1
S1, covering a monolayer of polystyrene microspheres on the surface of a substrate by a self-assembly method, wherein the diameter of the polystyrene microspheres is 100nm, the diameters of the polystyrene microspheres in the same batch are consistent, and the standard deviation of the particle sizes is less than 5%;
s2, placing the substrate covered with the polystyrene microspheres in a reactive ion etching device;
s3, using sulfur hexafluoride as a reaction gas, and performing reactive ion etching on the substrate surface where the polystyrene microspheres are located under the conditions of a gas flow rate of 30sccm, a cavity pressure of 15 Pa and an etching power of 150W until nanocones are formed on the surface of the substrate and the polystyrene microspheres are completely removed;
s4, depositing noble metal nanoparticles on the surface of the substrate where the nanocones are located by a magnetron sputtering method to form a noble metal film, and thus obtaining the common noble metal nanocone SERS substrate.
Example 1
S1, covering a monolayer of polystyrene microspheres on the surface of a substrate by a self-assembly method, wherein the diameter of the polystyrene microspheres is 100nm, the diameters of the polystyrene microspheres in the same batch are consistent, and the standard deviation of the particle sizes is less than 5%;
s2, smoothly bending the periphery of the substrate by 20 degrees towards the center of the surface where the polystyrene microspheres are located, and then placing the bent substrate in a reactive ion etching device;
s3, using sulfur hexafluoride as a reaction gas, and performing reactive ion etching on the substrate surface where the polystyrene microspheres are located under the conditions of a gas flow rate of 30sccm, a cavity pressure of 15 Pa and an etching power of 150W until nanocones are formed on the surface of the substrate and the polystyrene microspheres are completely removed;
s4, keeping the bending state of the substrate, depositing noble metal nanoparticles on the surface of the substrate where the nanocones are located through a magnetron sputtering method to form a noble metal film, and then flatting the bent substrate to obtain the noble metal nanocone SERS substrate for detecting the novel coronavirus.
Blowing 10 micrograms of polystyrene colloid microspheres with the diameter of 100nm at the gas flow rate of 100sccm, simultaneously capturing and collecting two precious metal nanocone SERS substrates prepared in a comparative example 1 and an embodiment 1, flushing with running water for 10 seconds, and then performing Raman spectrum characterization, as shown in figure 2, a curve I, II is respectively a Raman spectrum obtained by a common precious metal nanocone SERS substrate and a precious metal nanocone SERS substrate for detecting novel coronavirus, as can be seen from figure 2, the Raman spectrum obtained by the precious metal nanocone SERS substrate with different opening directions and opening sizes prepared by the preparation method disclosed by the invention has stronger characteristic peaks, and the higher capturing efficiency is shown.
It should be understood by those skilled in the art that the foregoing is only illustrative of several embodiments of the invention, and not of all embodiments. It should be noted that many variations and modifications are possible to those skilled in the art, and all variations and modifications that do not depart from the gist of the invention are intended to be within the scope of the invention as defined in the appended claims.
Claims (8)
1. The precious metal nanocone SERS substrate for detecting the novel coronavirus is characterized by comprising a substrate, a nanocone array positioned on the substrate and a precious metal film coated on the surface of the substrate where the nanocone array and the nanocone array are positioned, wherein the thickness of the precious metal film is 10-50nm, and the precious metal film is made of precious metal with Raman activity; the nano cone array comprises a plurality of nano cone rings with different diameters, the nano cone rings are formed by circumferentially arranging a plurality of nano cones around a nano cone perpendicular to a substrate, an included angle alpha formed between the central axis of each nano cone and the substrate is 30-90 degrees, the included angle alpha is gradually reduced from an inner ring of the nano cone ring to an outer ring of the nano cone ring, the height of each nano cone is 200-300nm, the diameter of the root of each nano cone is 50-300nm, the diameter of the tip of each cone is 20-50nm, the tip angle of each cone is 10-60 degrees, and the distance between every two adjacent nano cones is 80-150 nm.
2. The precious metal nanocone SERS substrate for detecting the novel coronavirus according to claim 1, wherein the substrate and the nanocone are made of the same flexible material and can be bent.
3. The precious metal nanocone SERS substrate for detecting the novel coronavirus according to claim 2, wherein the material of the substrate and the nanocone is one of polyimide and polyethylene terephthalate.
4. The precious metal nanocone SERS substrate for detecting the novel coronavirus according to claim 1, wherein the precious metal thin film is formed by stacking precious metal nanoparticles with the size of 10-30 nm.
5. The SERS substrate for detecting the novel coronavirus according to claim 4, wherein the material of the noble metal thin film is one of gold, silver and copper.
6. The precious metal nanocone SERS substrate for detecting a novel coronavirus according to claim 1, wherein the spacing between adjacent nanocones in the nanocone array is not completely equal or not completely equal.
7. A method for preparing the noble metal nanocone SERS substrate for detecting the novel coronavirus according to any one of claims 1 to 6, comprising the following steps:
s1, covering a monolayer of polystyrene microspheres on the surface of the substrate by a self-assembly method, wherein the diameter of the polystyrene microspheres is 80-150 nm;
s2, smoothly bending the periphery of the substrate towards the center of the surface where the polystyrene microspheres are located by 5-30 degrees, and then placing the bent substrate in a reactive ion etching device;
s3, using sulfur hexafluoride as a reaction gas, and performing reactive ion etching on the substrate surface where the polystyrene microspheres are located under the conditions that the gas flow rate is 10-50sccm, the cavity pressure is 5-20 Pa and the etching power is 100-;
s4, keeping the bending state of the substrate, depositing noble metal nanoparticles on the surface of the substrate where the nanocones are located through a magnetron sputtering method to form a noble metal film, and then flatting the bent substrate to obtain the noble metal nanocone SERS substrate for detecting the novel coronavirus.
8. The method for preparing the precious metal nanocone SERS substrate for detecting the novel coronavirus according to claim 7, wherein the diameters of the polystyrene microspheres in step S1 are not completely the same or different.
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| CN113046708A (en) * | 2021-02-09 | 2021-06-29 | 杭州电子科技大学 | Preparation method and application of magnetic field regulation and control nano array structure |
| CN113252640A (en) * | 2021-06-03 | 2021-08-13 | 季华实验室 | Rapid virus screening and detecting method |
| CN113702352A (en) * | 2021-08-25 | 2021-11-26 | 山东智微检测科技有限公司 | SERS detection chip suitable for gas-phase erosive toxicant and preparation method thereof |
| CN114199854A (en) * | 2021-12-15 | 2022-03-18 | 曲阜师范大学 | Preparation method of SERS substrate constructed by flexible transparent cone ordered array |
| CN115684580A (en) * | 2022-09-27 | 2023-02-03 | 郑州大学 | Magnetic control-based microsphere array chip, detection assembly, detection system and detection method |
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| CN113046708A (en) * | 2021-02-09 | 2021-06-29 | 杭州电子科技大学 | Preparation method and application of magnetic field regulation and control nano array structure |
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| CN115684580B (en) * | 2022-09-27 | 2023-08-25 | 郑州大学 | Microsphere array chip based on magnetic control, detection assembly, detection system and method |
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