CN113514427A - Biosensor for enhancing TORD spectrum detection and testing method - Google Patents
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Abstract
The invention discloses a biosensor for enhancing TORD spectrum detection, which comprises a terahertz wave refracting lens and a sub-wavelength metal sensing structure, wherein the terahertz wave refracting lens is in a hemispherical shape, the sub-wavelength metal sensing structure is a metal plate, a plurality of grids are uniformly distributed on the metal plate, the circular section of the terahertz wave refracting lens is arranged opposite to the gratings of the sub-wavelength metal sensing structure, and a gap to be detected is formed between the terahertz wave refracting lens and the sub-wavelength metal sensing structure. The biosensor has a simple structure, the test method is convenient and fast to operate, a locally enhanced terahertz electric field can be obtained in the sensing gap, the sample consumption is reduced, the influence of water absorption in the sample is overcome, the enhanced TORD spectrum measurement on trace biological samples (including solution samples) is realized, and the biosensor has an important application value in label-free sensing detection of the chiral biomolecules.
Description
Technical Field
The invention belongs to the technical field of biosensing, and particularly relates to a biosensor for enhancing TORD spectrum detection and a testing method.
Background
Terahertz (Terahertz, 1THz ═ 10)12Hz) wave refers to a wave with a frequency of 0.1THz-10 THzElectromagnetic waves with a length in the range of 0.03mm to 3mm, the frequency spectrum of which lies between the infrared and the microwave. Compared with traditional spectral analysis technologies such as infrared spectrum and X-ray, skeleton vibration, rotation spectrum and weak interaction force energy levels among molecules of a plurality of biological organic molecules such as protein, DNA, RNA, amino acid, sugar and the like are in a terahertz waveband, so that the terahertz spectrum technology has an important application prospect in the aspect of researching molecular characteristics of biological macromolecules.
Chirality is a fundamental property of nature and refers to the feature that an object and its mirror images cannot coincide. The structure of a chiral molecule does not have mirror symmetry itself, and the mirror projected molecule is the opposite enantiomer to its handedness. Most of the physical properties of chiral molecules that are enantiomers of each other are the same, but there is a significant difference in activity in organisms. Therefore, the qualitative and quantitative detection of chiral molecules has great significance in the fields of analytical science, chemical biology, medicines and the like.
At present, the common chiral detection methods are mainly divided into two types, chromatography and spectrometry. The chromatography mainly utilizes the selective adsorption capacity of a chromatographic column to chiral molecules for separation, purification and analysis. However, for different chiral molecules, the chromatography needs to select different chromatographic columns and mobile phases, which results in low universality, high operation difficulty and high analysis cost. Chiral spectroscopic techniques, such as Optical Rotation Dispersion (ORD), are currently an indispensable, fast and efficient method for studying molecular chirality based on the selective interaction between enantiomers and left and right circularly polarized electromagnetic waves to determine the absolute configuration and composition of the enantiomers.
Most biological macromolecules, such as proteins, DNA, RNA, etc., are sterically folded into structures with spatial chirality, which are true chiral molecules. The change of the collective vibration mode and the chiral advanced folding structure of chiral biomacromolecules such as protein and the like plays an important role in the life process, but the characteristic frequency of the chiral biomacromolecules is generally lower, and the chiral biomacromolecules are just in the terahertz frequency band. Therefore, the Terahertz Optical Rotation Dispersion (TORD) technology can provide more sensitive and efficient detection and analysis for identification of chiral biomacromolecules and interaction between molecules.
However, the optical activity response of chiral samples such as biomolecules in a terahertz wave band is weak, so that direct and effective detection is difficult, and a mature TORD measuring scheme does not exist at present. The surface plasmon is a surface wave with high local area on the metal surface, is very sensitive to the dielectric environment of the metal surface, and can be used as a biosensor. The electromagnetic field of the surface plasmon has the characteristic of great electromagnetic enhancement on the surface of the sensing metal, and can enhance the interaction between incident waves and surface substances and amplify the optical activity response of the surface substances. The invention utilizes the designed sensor structure to scan the incident angle to excite the surface plasmon resonance with different frequencies, and utilizes the electromagnetic enhancement effect to obtain the broadband enhanced TORD spectrum of the sample, thereby realizing the TORD spectrum sensing analysis of the trace chiral sample.
Disclosure of Invention
The invention aims to provide a biosensor for enhancing TORD spectrum detection and a testing method thereof, which utilize the electromagnetic enhancement effect of surface plasmon excited in a sub-wavelength metal sensing layer to increase the interaction between an incident wave and a sample, amplify the optical activity response of the sample, obtain the enhancement response effect near different frequency points by scanning the incident angle, and realize the broadband TORD spectrum detection of the sample.
The invention solves the technical problem by adopting the scheme that the biosensor for enhancing the TORD spectrum detection comprises a terahertz wave refractive lens and a sub-wavelength metal sensing structure, wherein the terahertz wave refractive lens is in a hemispherical shape, the sub-wavelength metal sensing structure is a metal plate, a plurality of grids are uniformly distributed on the metal plate, the circular section of the terahertz wave refractive lens is arranged opposite to the grids of the sub-wavelength metal sensing structure, and a gap to be detected is formed between the terahertz wave refractive lens and the sub-wavelength metal sensing structure.
Furthermore, the terahertz wave refracting lens is made of a terahertz wave band high-refractive-index material, and is one of high-resistance silicon, high-resistance germanium and methyl pentene polymer.
Further, the spacing size of the grating of the sub-wavelength metal sensing structure is in the sub-wavelength range, namely, smaller than the wavelength of the terahertz wave, and the size is between 5 micrometers and 900 micrometers.
Furthermore, the sub-wavelength metal sensing structure is a one-dimensional periodic structure or a two-dimensional periodic structure.
Furthermore, the size of the distance between the gaps to be measured can be adjusted, the distance is in a sub-wavelength range, namely, the size of the distance is smaller than the wavelength of incident terahertz waves, and the distance is between 5 micrometers and 900 micrometers.
Preferably, the chiral sample to be detected in the biosensor can be, but is not limited to, a solution sample, a gas sample, and a powder sample, and the chiral sample to be detected is filled in the gap to be detected, and can also be deposited on the surface of the sub-wavelength metal sensing structure.
The invention also provides a test method of the biosensor for enhancing the TORD spectrum detection by adopting the technology, which is characterized by comprising the following steps of filling an article to be detected in a gap to be detected, utilizing a terahertz wave generator to inject terahertz waves into a terahertz wave refracting lens of the biosensor, wherein the incident terahertz waves are transverse magnetic polarized waves, the angle of the incident waves is larger than the total reflection critical angle of the hemispherical column lens material, the incident waves generate total reflection on the bottom surface of the hemispherical column lens, corresponding evanescent waves are formed above the bottom surface, the generated evanescent waves are coupled and excited to form a surface plasmon resonance phenomenon on a subwavelength metal sensing structure, an electromagnetic enhancement effect is generated, so that the interaction between the incident waves and a filled chiral sample is increased, and the optical activity response difference of the chiral sample to left and right circular polarization components in the incident waves is amplified, after the action, the terahertz signal is emitted from the other side of the hemispherical cylindrical lens to be detected; detecting two orthogonal components of the emergent signal to obtain reflection spectrums of left and right circular polarization components, wherein the difference of the reflection spectrums is a TORD signal, and the TORD signal with narrow-band enhancement near the resonance frequency of the surface plasmon can be obtained by utilizing the electromagnetic enhancement effect of the surface plasmon; and thirdly, exciting the surface plasmon resonance enhancement effect under different frequencies by scanning different incidence angles, enhancing the optical activity response of the sample under different frequencies, integrating and extracting narrow-band TORD signals corresponding to different incidence angles to obtain a broadband enhanced TORD signal, and realizing the enhanced TORD spectrum sensing analysis of the chiral sample to be detected.
Compared with the prior art, the invention has the following beneficial effects:
1. the biosensor combines the sub-wavelength metal sensing structure and the hemispherical cylindrical lens to form a certain tiny gap as a sample containing pool, reduces the sample consumption and is suitable for the detection of trace samples.
2. The surface plasmon resonance on the metal sensing layer structure is efficiently excited by utilizing the incident terahertz wave, the optical activity response of the tested sample is amplified by utilizing the electromagnetic enhancement effect, the detection sensitivity is improved, and the enhanced TORD spectrum of the sample can be obtained.
3. The method comprises the steps of exciting surface plasmon resonances of different frequency points by scanning an incident angle of terahertz waves to obtain enhanced TORD spectrums near the different frequency points, and obtaining broadband enhanced TORD spectrums of a sample through data integration.
4. The sensor has a simple structure, the testing method is convenient and fast to operate, a locally enhanced terahertz electric field can be obtained in the sensing gap, the using amount of a sample is reduced, the influence of water absorption in the sample is overcome, the enhanced TORD spectrum measurement on trace biological samples (including solution samples) is realized, and the sensor has an important application value in label-free sensing detection of the chiral biomolecules.
Drawings
FIG. 1 is a schematic structural diagram of the present invention.
Fig. 2 is a schematic diagram of the operation of the sensor of the present invention.
FIG. 3 is a TORD spectrum of a chiral sample to be measured obtained by using the sensor of the present invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings.
As shown in fig. 1, a biosensor capable of obtaining a broadband enhanced terahertz absorption spectrum includes a terahertz wave refractive lens 1 and a sub-wavelength metal sensing structure 2, wherein the terahertz wave refractive lens 1 is shaped like a hemisphere, the sub-wavelength metal sensing structure 2 is a metal plate 21, a plurality of grids 22 are uniformly distributed on one surface of the metal plate 21, a circular section of the terahertz wave refractive lens 1 is arranged opposite to a grid surface of the sub-wavelength metal sensing structure 2, and a gap 3 to be measured is formed between the terahertz wave refractive lens 1 and the sub-wavelength metal sensing structure 2. The terahertz wave refracting lens 1 is made of a terahertz wave band high-refractive-index material, and is one of high-resistance silicon, high-resistance germanium and methyl pentene polymers. The spacing size of the grating of the sub-wavelength metal sensing structure is in a sub-wavelength range, namely smaller than the wavelength of terahertz waves, and the size is between 5 micrometers and 900 micrometers. The size of the distance between the gaps to be measured can be adjusted, the distance is in a sub-wavelength range, namely, the size of the distance is smaller than the wavelength of incident terahertz waves, and the distance is between 5 micrometers and 900 micrometers.
The test method of the biosensor capable of acquiring the broadband enhanced terahertz absorption spectrum, which adopts the technical scheme, comprises the following steps: filling an object to be detected in the gap to be detected, injecting terahertz waves into a terahertz wave refracting lens of the biosensor by using a terahertz wave generator, wherein the incident terahertz waves are transverse magnetic polarized waves, the angle of the incident waves is larger than the total reflection critical angle of the hemispherical cylindrical lens material, the incident waves generate total reflection on the bottom surface of the hemispherical cylindrical lens, forming corresponding evanescent wave above the bottom surface, coupling the generated evanescent wave to excite the surface plasmon on the subwavelength metal sensing structure to form surface plasmon resonance phenomenon and generate electromagnetic enhancement effect, therefore, the interaction between the incident wave and the filled chiral sample is increased, the difference of the optical activity response of the chiral sample to left and right circularly polarized components in the incident wave is amplified, and the terahertz signal is emitted from the other side of the hemispherical cylindrical lens to be detected after the interaction; detecting two orthogonal components of the emergent signal to obtain reflection spectrums of left and right circular polarization components, wherein the difference of the reflection spectrums is a TORD signal, and the TORD signal with narrow-band enhancement near the resonance frequency of the surface plasmon can be obtained by utilizing the electromagnetic enhancement effect of the surface plasmon; and thirdly, exciting the surface plasmon resonance enhancement effect under different frequencies by scanning different incidence angles, enhancing the optical activity response of the sample under different frequencies, integrating and extracting narrow-band TORD signals corresponding to different incidence angles to obtain a broadband enhanced TORD signal, and realizing the enhanced TORD spectrum sensing analysis of the chiral sample to be detected.
As shown in fig. 2, the straight line (dashed line) in the figure is the vacuum wave vector, and the straight line (solid line) is the horizontal wave vector of the incident wave in the cylindrical lens under the incident of total reflection. In the figure, the curves (solid lines and dotted lines) are wave vector curves corresponding to left and right circularly polarized components of the plasmon on the upper surface of the sensing layer of the sub-wavelength metal grating structure when the filled sample has no chirality (k is 0), and it can be seen that the wave number curves of the left and right circularly polarized components are coincident in this case. Curves (dotted line, two-dot line) in the figure are wave vector curves of left and right circularly polarized components when the filled sample to be tested has chirality (k > 0). It can be seen that after filling the chiral sample, the two appear separated due to the different response indices of the different circularly polarized components to the chiral sample. When the filled sample to be tested has opposite chirality (k <0), the left and right circular polarization component curves will exchange positions.
As can be seen in the figure, the wave vector of the incident wave in the prism and the wave vector curve of the surface plasmon on the sensing layer have an intersection point, and the intersection point corresponds to the intersection point which is formed by coupling the incident wave and exciting the surface plasmon resonance. Under the condition of filling a chiral sample, the excited left and right circularly polarized surface plasmon resonance frequency positions are separated, and the separation difference is positively correlated with the chiral parameter size of the sample, so that the TORD sensing signal can be obtained.
Changing the angle of incidence changes the frequency point at which the excited surface plasmon resonates. By scanning the incident angle, the surface plasmon resonance can be excited in a broadband range, and broadband enhanced TORD spectrum detection of the sample is realized.
FIG. 3 is a comparison graph of enhanced TORD spectra obtained with the inventive sensor. The solid and dashed lines in the figure correspond to the left and right circularly polarized reflectance spectra when filled with achiral samples (k 0). It can be seen that when the sample does not have chirality, both are the same. In the figure, the dashed and dotted lines correspond to left and right circularly polarized component reflectance spectra of a filled chirally-sized sample (k ═ 0.1), respectively. The dot-dash line and the two-dot-dash line in the figure correspond to left and right circularly polarized component reflection spectra of a filled chirally large sample (k 0.2), respectively. The minimum valley in the reflection spectrum is the surface plasmon resonance excitation position. As can be seen from the figure, after the chiral sample is filled, the resonance frequencies of the excited left and right circularly polarized surface plasmon elements are separated, and the separation degree is positively correlated with the chiral size of the sample, so that the TORD spectrum detection of the sample is realized. By scanning different incidence angles and changing the excitation frequency of the surface plasmon, the broadband enhanced TORD spectrum information of the sample can be obtained.
The above are preferred embodiments of the present invention, and all changes made according to the technical scheme of the present invention that produce functional effects do not exceed the scope of the technical scheme of the present invention belong to the protection scope of the present invention.
Claims (7)
1. A biosensor for enhancing the detection of TORD spectrum, which is characterized in that: the terahertz wave sensor comprises a terahertz wave refracting lens and a sub-wavelength metal sensing structure, wherein the terahertz wave refracting lens is in a hemispherical shape, the sub-wavelength metal sensing structure is a metal plate, a plurality of grids are uniformly distributed on the metal plate, a circular section of the terahertz wave refracting lens and the gratings of the sub-wavelength metal sensing structure are arranged oppositely, and a gap to be measured is formed between the terahertz wave refracting lens and the sub-wavelength metal sensing structure.
2. The biosensor for enhancing the TORD spectroscopy of claim 1, wherein: the terahertz wave refracting lens is made of a terahertz wave band high-refractive-index material and is one of high-resistance silicon, high-resistance germanium and methyl pentene polymers.
3. The biosensor for enhancing the TORD spectroscopy of claim 1, wherein: the spacing size of the grating of the sub-wavelength metal sensing structure is in a sub-wavelength range, namely smaller than the wavelength of terahertz waves, and the size is between 5 micrometers and 900 micrometers.
4. The biosensor for enhancing the TORD spectroscopy of claim 1, wherein: the sub-wavelength metal sensing structure is a one-dimensional periodic structure or a two-dimensional periodic structure.
5. The biosensor for enhancing the TORD spectroscopy of claim 1, wherein: the size of the distance between the gaps to be measured can be adjusted, the distance is in a sub-wavelength range, namely, the size of the distance is smaller than the wavelength of incident terahertz waves, and the distance is between 5 micrometers and 900 micrometers.
6. The biosensor for enhancing the TORD spectroscopy of claim 1, wherein: the chiral sample to be detected in the biosensor can be, but is not limited to, a solution sample, a gas sample and a powder sample, and the chiral sample to be detected is filled in the gap to be detected and can also be deposited on the surface of the sub-wavelength metal sensing structure.
7. The method as claimed in claim 1, wherein the method comprises the steps of filling an object to be tested in the gap, injecting terahertz waves into the terahertz wave refractive lens of the biosensor by using the terahertz wave generator, wherein the incident terahertz waves are transverse magnetic polarized waves, the angle of the incident waves is larger than the critical angle of total reflection of the hemispherical cylindrical lens material, the incident waves are totally reflected at the bottom surface of the hemispherical cylindrical lens, corresponding evanescent waves are formed above the bottom surface, the generated evanescent waves are coupled and excited to surface plasmon resonance on the subwavelength metal sensing structure, so as to form surface plasmon resonance, generate electromagnetic enhancement effect, thereby increasing the interaction between the incident waves and the filled chiral sample, and amplifying the difference of optical activity response of the chiral sample to left and right circularly polarized components in the incident waves, after the action, the terahertz signal is emitted from the other side of the hemispherical cylindrical lens to be detected; detecting two orthogonal components of the emergent signal to obtain reflection spectrums of left and right circular polarization components, wherein the difference of the reflection spectrums is a TORD signal, and the TORD signal with narrow-band enhancement near the resonance frequency of the surface plasmon can be obtained by utilizing the electromagnetic enhancement effect of the surface plasmon; and thirdly, exciting the surface plasmon resonance enhancement effect under different frequencies by scanning different incidence angles, enhancing the optical activity response of the sample under different frequencies, integrating and extracting narrow-band TORD signals corresponding to different incidence angles to obtain a broadband enhanced TORD signal, and realizing the enhanced TORD spectrum sensing analysis of the chiral sample to be detected.
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Cited By (2)
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| CN114324241A (en) * | 2022-01-06 | 2022-04-12 | 上海理工大学 | Sensor based on pseudo surface plasmon three-dimensional stacking structure |
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