CN114199828B - Metal-graphene-based hybrid super-surface biosensor and preparation method thereof - Google Patents
Metal-graphene-based hybrid super-surface biosensor and preparation method thereof Download PDFInfo
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- CN114199828B CN114199828B CN202111492145.4A CN202111492145A CN114199828B CN 114199828 B CN114199828 B CN 114199828B CN 202111492145 A CN202111492145 A CN 202111492145A CN 114199828 B CN114199828 B CN 114199828B
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Abstract
The invention discloses a metal-graphene-based hybrid super-surface biosensor and a preparation method thereof. The sensor takes an ultrathin flexible Polyimide (PI) film as a substrate, a periodically arranged metal grating structure is prepared on the substrate, an alumina thin layer is deposited on the surface of the metal grating, and a single-layer graphene layer is paved above the alumina thin layer which is opposite to the structure. When terahertz waves are normally incident to the surface of the structure, graphene surface plasmon polaritons (GSPs) resonance effects and abnormal optical transmission (EOT) resonance in the metal grating are excited. The sensor of the invention is based on the super surface design of strong coupling of EOT resonance and GSPs resonance, and the sensitivity of the sensor is S Ω =1.77 THz/eV. The ultrathin flexible PI substrate not only can reduce electromagnetic loss, but also has good ductility, is suitable for a curved surface, and is convenient for measurement of a sample. The biosensor has the advantages of low preparation cost, good ductility, high sensitivity, convenient specific detection and measurement, etc.
Description
Technical Field
The invention relates to a terahertz biosensor for label-free bioprotein detection and high-sensitivity specific detection of a pi-electron-containing bioprotein, in particular to a terahertz super-surface biosensor based on strong coupling between abnormal optical transmission (EOT) in a sub-wavelength metal grating and Graphene Surface Plasmons (GSPs) and a preparation method thereof.
Background
Terahertz (THz) is an electromagnetic spectrum between microwave/millimeter wave and infrared radiation with frequencies in the range of 100GHz to 10THz. Terahertz waves have many excellent characteristics in the biomedical field: (1) The photon energy of terahertz radiation is very low (1 THz about 4 meV), avoiding ionizing damages to biological samples like X-rays; (2) The terahertz has longer wavelength compared with the visible light and infrared wave bands, so the terahertz is not influenced by Mie scattering, and the terahertz is suitable for nondestructive detection of samples;
(3) The vibration and rotation energy spectrum of many molecules is in the terahertz range, so that the component information of the detected object can be identified by using the terahertz. Terahertz research is also faced with challenges, however, such as the lack of high-power terahertz sources and high-sensitivity terahertz detectors. In recent years, as the performance of terahertz functional devices is improved, research on strong interactions of terahertz with substances has attracted tremendous attention from researchers. The existing commonly used terahertz biological sample detection methods and devices are terahertz spectrum analysis, split Ring Resonator (SRRs) metamaterial sensing, metamaterial perfect absorber (PMAs) sensing, terahertz microfluidic sensors and the like. However, these related sensing methods have certain limitations, such as: terahertz spectrum detection sensitivity is low. The SRRs sensor and the terahertz waves generate single interaction to form low resonance intensity, strong local field enhancement cannot be generated, and the interaction between the object to be detected and the local field is weak, so that the quality factor Q and the sensitivity of the SRRs are low. When the current PMAs terahertz sensor detects substances, the substances to be detected are positioned outside the Fabry-Perot resonant cavity and attached to the surface of the periodic layer of the metal structure, and cannot be overlapped with the space of the local enhanced electric field, so that only weak interaction is generated between the substances to be detected and the periodic layer of the metal structure, and the sensitivity of the sensor is low. When the terahertz microfluidic channel sensor detects, due to the strong absorption characteristic of water to terahertz waves, the incident terahertz waves can be weakened by adding liquid into the fluid channel, so that the resonance intensity formed by interaction of the sensor and the terahertz waves is low, and the detection effect is influenced. And none of these detection means is currently capable of specifically detecting biological samples.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the metal-graphene hybrid super-surface biosensor based on the strong coupling effect and the preparation method thereof, wherein the addition of the analyte on the surface of the graphene can cause the change of the doping condition of the graphene, thereby causing the change of the carrier concentration in the graphene. The change in carrier concentration of graphene in the structure can result in a change in the strong coupling spectrum. Thus, we can detect analytes through frequency shifts of spectral resonances. Because the graphene contains delocalized pi electrons, the pi-electron-containing analyte can be selectively adsorbed in a pi-pi bond stacking mode, the adsorption efficiency of the analyte is further improved, and the Fermi energy level of the p-type doped graphene is led to the Dirac point, so that the highly sensitive specific detection of pi-electron-containing substances is realized.
In order to solve the technical problems, the invention adopts the following technical scheme: a Rabi-split metal-graphene hybrid super-surface biosensor based on a strong coupling effect takes an ultrathin flexible PI film as a substrate, sub-wavelength metal grating structures are periodically distributed on the surface of the substrate, and the period (p) and the gap (w) are 178 mu m and 42 mu m respectively. An approximately 30nm thick film of aluminum oxide was deposited on the surface of the metal strips. And a layer of monolayer graphene is paved above the alumina thin layer which is opposite to the structure.
Furthermore, the metal grating is of a copper structure and is prepared on the PI substrate through an electron beam evaporation coating technology, a laser direct writing technology and a reactive ion etching technology.
A preparation method of a metal-graphene hybrid super-surface biosensor is provided, and graphene realizes high-sensitivity specific detection on a substance containing pi electrons in a manner that the Fermi level of p-type doped graphene is led to a Dirac point through pi-pi bond accumulation. The specific preparation process comprises the following steps:
s1, bonding a flexible PI film with the thickness of 50 mu m on the surface of a flat silicon wafer by utilizing the thermosetting property of Polydimethylsiloxane (PDMS);
s2, plating metal copper with the thickness of 100-300nm on the surface of the PI film by utilizing an electron beam evaporation coating system;
s3, uniformly spin-coating a layer of positive photoresist with the thickness of 0.7-3.0 mu m on the PI film subjected to copper plating through a spin coater, and exposing and developing by using a laser direct writing system;
s4, etching the developed structure through a reactive ion etching technology;
s5, depositing an alumina film with the thickness of about 30nm on the surface of the metal strip by using an atomic layer deposition technology;
s6, stripping the structure from the silicon wafer and attaching the structure to a hollowed PET film with the thickness of 100-300 mu m so as to fix PI;
s7, transferring the graphene film to the surface of alumina through wet transfer, overlapping the graphene and a metal grating area up and down in the process of fishing out the graphene, and then dissolving the PMMA coating supporting the graphene with acetone for three times.
According to the metal-graphene hybrid super-surface-based biosensor, an ultrathin flexible PI film is used as a substrate, and periodically arranged metal grating structures are prepared on the substrate. And depositing a 30nm thick low-dielectric-constant alumina dielectric layer on the metal grating, and paving a single-layer graphene on the alumina layer on the grating. When terahertz waves are normally incident, GSPs effect and EOT of the metal grating are excited. This special design can cause the EOT effect of the metal grating to interact with GSPs, resulting in a Rabi splitting with strong coupling effect. The addition of analytes to the graphene surface causes a change in the doping profile of the graphene, resulting in a change in carrier concentration in the graphene. The change in carrier concentration of graphene in the structure can result in a change in the strong coupling spectrum. Thus, we can detect analytes through frequency shifts of spectral resonances. Because the graphene contains delocalized pi electrons, the pi-electron-containing analyte can be selectively adsorbed in a pi-pi bond stacking mode, the adsorption efficiency of the analyte is further improved, and the Fermi energy level of the p-type doped graphene is led to the Dirac point, so that the highly sensitive specific detection of pi-electron-containing substances is realized.
Compared with the prior art, the invention has the following advantages:
1. the invention provides a metal-graphene hybrid super-surface biosensor, which uses ultrathin flexible PI with low-loss dielectric property as a substrate, so that the electromagnetic loss can be reduced, the metal-graphene hybrid super-surface biosensor has good ductility, is suitable for a curved surface, and is convenient for measuring a sample. In addition, compared with the terahertz spectrum analysis commonly used at present, the sensor designed by the invention is a terahertz biological sample detection method and device such as a Split Ring Resonator (SRRs) metamaterial sensing, a metamaterial perfect absorber (PMAs) sensing, a terahertz microfluidic sensor and the like. The sensor has the advantages of rapidness, sensitivity, low cost, good ductility, small volume, convenient measurement and the like, and can be widely applied to the field of biological sensing.
2. The metal-graphene hybrid super-surface biosensor provided by the invention has the advantages that the designed metal grating and the whole graphene can cause the interaction of EOT resonance of the metal grating and GSPs resonance effect when terahertz waves are normal incidence, the Rabi splitting with strong coupling effect is generated, the sensitivity is higher, and the sensitivity of the sensor obtained by fitting is S Ω =1.77 THz/eV. According to the sensor, graphene is used as a sensing contact surface, so that small change of carrier concentration in the graphene can be detected, delocalized pi electrons in the graphene can be utilized, and the fermi level of p-type doped graphene is led to a Dirac point in a pi-pi bond stacking mode, so that high-sensitivity specific detection of a substance containing pi electrons is realized. Test results show that the terahertz super-surface biosensor can realize high-sensitivity specific detection of the protein containing pi electrons, and provides a new feasible way for further application of biosensing in the terahertz field.
Drawings
FIG. 1 is a flow chart of the fabrication of a subsurface biosensor structure according to the present invention.
FIGS. 2 (a) and (b) are a physical diagram and a schematic structural diagram of the biosensor, respectively.
Fig. 3 is a transmission spectrum of the biosensor when graphene correspondence of different fermi energies matches resonant metal grating parameters. (the relevant parameters are marked in the upper right corner of the figure)
Fig. 4 (a) and (b) show simulation results of the variation of the strongly coupled transmission spectrum with the fermi energy and the sensitivity of the splitting degree Ω to the fermi energy, respectively.
FIG. 5 is a fermi energy E of the present invention F From-0.18 eV to-0.Calculated transmission spectra for different fermi levels of 26eV, E F The negative sign of (a) indicates that the graphene used is hole doped.
Detailed Description
The invention is described in further detail below with reference to the drawings and examples.
The different fermi energies are different for the metal structure parameters that can undergo strong coupling symmetric splitting, fig. 3 shows the Rabi splitting transmission spectrum of the biosensor when the graphene correspondence of the different fermi energies matches the resonant metal grating parameters, and the relevant parameters are marked in the upper right corner of the figure. Fig. 4 (a) shows the transmission spectrum change of the hybrid structure corresponding to the fermi energy of graphene from-0.18 eV to-0.26 eV when the metal grating structure parameter is p=178 μm and w=42 μm, and it can be seen from the figure that the Rabi frequency is gradually increased as the fermi energy is increased. When the fermi level is below/above-0.2 eV, the resonance peak will appear red-shifted/blue-shifted. Coupling weakens as the resonance frequency moves away from-0.2 eV and eventually causes the splitting to vanish. The sensitivity of the hybrid subsurface to fermi energy can be assessed by detecting a change in the strongly coupled Rabi frequency Ω. The sensitivity of the strongly coupled sensor is defined as S Ω ＝ΔΩ/ΔE F . Where ΔΩ represents the Rabi frequency variation due to the fermi energy variation. As can be seen from FIG. 4 (b), S Ω The Fermi energy sensitivity of the device is obtained by fitting the linear increase along with the increase of the Fermi energy Ω ＝1.77THz/eV。
As shown in fig. 2 (b), a metal-graphene hybrid-based super-surface biosensor uses an ultrathin flexible PI film as a substrate, and sub-wavelength metal grating structures are periodically arranged on the surface of the substrate, wherein the period (p) and the gap (w) are 178 μm and 42 μm respectively. An approximately 30nm thick film of aluminum oxide was deposited on the surface of the metal strips. And a layer of monolayer graphene is paved above the alumina thin layer which is opposite to the structure.
Furthermore, the metal grating is of a copper structure and is prepared on the PI substrate through an electron beam evaporation coating technology, a laser direct writing technology and a reactive ion etching technology.
According to the preparation method of the metal-graphene hybrid super-surface biosensor, the graphene enables the Fermi level of p-type doped graphene to be led to the Dirac point through pi-pi bond accumulation, so that high-sensitivity specific detection on the biological protein containing pi electrons can be realized. As shown in fig. 1, the specific preparation process comprises the following steps:
s1, bonding a flexible PI film with the thickness of 50 mu m on the surface of a flat silicon wafer by utilizing the thermosetting property of Polydimethylsiloxane (PDMS);
s2, plating metal copper with the thickness of 100-300nm on the surface of the PI film by utilizing an electron beam evaporation coating system;
s3, uniformly spin-coating a layer of positive photoresist with the thickness of 0.7-3.0 mu m on the PI film subjected to copper plating through a spin coater, and exposing and developing by using a laser direct writing system;
s4, etching the developed structure through a reactive ion etching technology;
s5, depositing an alumina film with the thickness of about 30nm on the surface of the metal strip by using an atomic layer deposition technology;
s6, stripping the structure from the silicon wafer and attaching the structure to a hollowed PET film with the thickness of 100-300 mu m so as to fix PI;
s7, transferring the graphene film to the surface of alumina through wet transfer, overlapping the graphene and a metal grating area up and down in the process of fishing out the graphene, and then dissolving the PMMA coating supporting the graphene with acetone for three times.
The structure produced is shown in FIG. 2 (a).
In this embodiment, when the terahertz wave is normally incident to the surface of the structure, the GSPs resonance effect and the EOT effect in the metal grating are excited. This special design can cause the EOT of the metal grating to interact with the GSPs in resonance, resulting in Rabi splitting with strong coupling effect. The addition of analytes to the graphene surface causes a change in the doping profile of the graphene, resulting in a change in carrier concentration in the graphene. The change in carrier concentration of graphene in the structure can result in a change in the strong coupling spectrum. Thus, we can detect analytes through frequency shifts of spectral resonances. Because the graphene contains delocalized pi electrons, the pi-electron-containing analyte can be selectively adsorbed in a pi-pi bond stacking mode, the adsorption efficiency of the analyte is further improved, and the Fermi energy level of the p-type doped graphene is led to the Dirac point, so that the highly sensitive specific detection of pi-electron-containing substances is realized. The method for detecting the biological protein based on the metal-graphene hybrid super-surface biological sensor provided by the invention comprises the following steps:
the first step: and removing organic matters on the surface of the prepared super-surface biosensor by adopting a method of cleaning with acetone and then ethanol, then washing with deionized water, and finally drying with nitrogen. And (3) measuring THz time domain spectrum signals in a dry air environment (high-purity nitrogen is continuously introduced) through a THz-TDs system of a laboratory, and obtaining transmission frequency domain spectrum data of the blank structure super surface through Fourier transformation.
And a second step of: aβ (16-22) polypeptide was prepared as a 1mg/mL solution with ultrapure water (pH=7.1), and incubated in a 37℃water bath for 0.5 hours, 1 hour and 1.5 hours, respectively, to obtain Aβ (16-22) polypeptide samples in different growth states. 10 mu L of polypeptide solution is dripped on the surface of the structure each time. And (3) measuring THz time domain spectrum signals in a dry air environment (high-purity nitrogen is continuously introduced) through a THz-TDs system, and obtaining transmission frequency domain spectrum data of the super surface of the Abeta (16-22) polypeptide at different culture times through Fourier transformation.
And a third step of: THz time domain spectrum signal measurement is carried out in a dry air environment (high-purity nitrogen is continuously introduced) through a THz-TDs system of a laboratory, and transmission frequency domain spectrum data of air is obtained through Fourier transformation.
Fourth step: and (3) carrying out arrangement analysis on all obtained measurement data results, and respectively drawing corresponding transmission spectrograms.
When Abeta (16-22) polypeptides with different incubation times are added, as shown in FIG. 5, a spectrum of frequency domain transmission coefficients is obtained through Fourier transform, and the transmission rate is defined as T= |E sample /E reference | 2 Wherein E is sample And E is reference Pulse information for the subsurface sample and nitrogen, respectively. The transmission spectrum after treatment is shown in FIG. 5 (a), and it can be seen that there is a significant shift in the transmission spectrum as Aβ (16-22) grows. Compared with the super without dripping the polypeptideThe surface transmission response (Ω is 232 GHz), the monomeric polypeptide causes a red shift in the strongly coupled splitting of the transmission spectrum, while Ω decreases from 232GHz to 171GHz (ΔΩ=61 GHz). The introduction of the oligomer resulted in splitting of Ω down from 232GHz to 191GHz (ΔΩ=41 GHz), whereas the polypeptide in fibrous form resulted in Ω changing from 232GHz to 228GHz (ΔΩ=4 GHz). Fig. 5 (b) summarizes the response of Rabi frequency Ω with aβ (16-22) morphology, indicating that we propose a metal-graphene hybrid subsurface that can be used to successfully detect aβ (16-22) fibrosis using a terahertz spectroscopy system.
The method has the advantages that the Abeta (16-22) polypeptide in different growth stages is dripped on the hybrid super surface and detected by using a terahertz time-domain spectroscopy system, and experimental results show that the method can rapidly and intuitively detect the polypeptide in different growth states. This work has enabled the use of the structure in the field of biosensing.
The above description is only of the preferred embodiments of the present invention, and is not intended to limit the present invention. Any simple modification, variation and equivalent variation of the above embodiments according to the technical substance of the invention still fall within the scope of the technical solution of the invention.
Claims (2)
1. A preparation method of a metal-graphene hybrid super-surface biosensor is characterized in that graphene leads the fermi level of p-type doped graphene to a dirac point through pi-pi bond accumulation to realize high-sensitivity specific detection on a pi-electron-containing substance; the specific preparation process comprises the following steps:
s1, combining a flexible PI film with the thickness of 50 mu m on the surface of a flat silicon wafer by utilizing the thermosetting property of polydimethylsiloxane PDMS;
s2, plating metal copper with the thickness of 100-300nm on the surface of the PI film by utilizing an electron beam evaporation coating system;
s3, uniformly spin-coating a layer of positive photoresist with the thickness of 0.7-3.0 mu m on the PI film subjected to copper plating through a spin coater, and exposing and developing by using a laser direct writing system;
s4, etching the developed structure through a reactive ion etching technology;
s5, depositing an alumina film with the thickness of about 30 and nm on the surface of the metal strip by using an atomic layer deposition technology;
s6, stripping the structure from the silicon wafer and attaching the structure to a hollowed PET film with the thickness of 100-300 mu m so as to fix PI;
s7, transferring the graphene film to the surface of alumina through wet transfer, overlapping the graphene and a metal grating area up and down in the process of fishing out the graphene, and then dissolving the PMMA coating supporting the graphene with acetone for three times.
2. The metal-graphene hybrid-based super-surface biosensor prepared by the preparation method of claim 1 is characterized in that an ultrathin flexible PI film is used as a substrate, a sub-wavelength metal grating structure is periodically arranged on the surface of the substrate, and the period p and the gap w are 178 mu m and 42 mu m respectively; depositing an aluminum oxide film about 30 a nm a thick on the surface of the metal strips; and a layer of monolayer graphene is paved above the alumina thin layer which is opposite to the structure.
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| US9423345B2 (en) * | 2014-06-24 | 2016-08-23 | International Business Machines Corporation | Chemical sensors based on plasmon resonance in graphene |
| US10877194B2 (en) * | 2018-01-16 | 2020-12-29 | National Technology & Engineering Solutions Of Sandia, Llc | Tunable graphene-based infrared reflectance filter having patterned nanoantenna layer and unpatterned graphene layer |
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| CN104466617A (en) * | 2013-09-18 | 2015-03-25 | 中国科学院苏州纳米技术与纳米仿生研究所 | Terahertz light source chip and manufacturing method thereof, terahertz light source device and manufacturing method thereof, and terahertz light source module and manufacturing method thereof |
| CN112939478A (en) * | 2021-01-14 | 2021-06-11 | 南昌大学 | Preparation method of porous film material for shielding terahertz waves |
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