CN111307755A - Method for detecting liquid sample based on terahertz technology - Google Patents
Method for detecting liquid sample based on terahertz technology Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3581—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using far infrared light; using Terahertz radiation
- G01N21/3586—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using far infrared light; using Terahertz radiation by Terahertz time domain spectroscopy [THz-TDS]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3577—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing liquids, e.g. polluted water
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Abstract
The invention provides a method for detecting a liquid sample based on a terahertz technology, which comprises the following steps of S1: providing a terahertz spectrum device, a container capable of containing a liquid sample, and a metamaterial sensor made of a terahertz metamaterial, wherein the metamaterial sensor is arranged close to the inner wall of one side of the container; s2: preparing a biomolecule to be detected into reverse micelle emulsion, and adding the reverse micelle emulsion into a container, wherein the reverse micelle emulsion is an emulsion with a continuous phase as an oil phase, a dispersed phase as a water phase and phospholipid molecules as a surfactant, the phospholipid molecules exist in a reverse micelle form, and the biomolecule to be detected is wrapped in the dispersed phase; and S3: and a terahertz spectrum device is used for obtaining the spectrum information of the total body to be detected in the terahertz wave band so as to realize the detection of the liquid sample. The invention utilizes a terahertz spectrum device, and based on the combination of a terahertz metamaterial and a reverse micelle, detects the property and interaction of biomolecules in a microenvironment and phospholipid self-assembly mode under the condition close to the physiological condition and carries out quantitative detection.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to a method for detecting a liquid sample based on a terahertz technology.
Background
Terahertz is generally an electromagnetic wave with a frequency of 0.3 to 10THz and a wavelength of 30 μm to 1 mm. Located in the microwave in the electromagnetic spectrumAnd the infrared band. The terahertz time-domain spectroscopy (THz-TDS) technology is a powerful tool for researching dielectric relaxation of water molecules at biological interface, and the time measurement scale range of the THz technology is 10-9To 10-11S, direct information capable of characterizing picosecond hydration kinetics is an ideal tool for researching bound water. This means that the THz band has unique advantages for use in interfacial water environments and organic biomolecules (e.g. DNA, proteins, etc.). Currently, the application of terahertz technology to analyze organic biological samples has become an important direction for the development of terahertz technology. The development of the terahertz time-domain spectroscopy technology opens a brand new visual field for people to research biomolecules, and the high sensitivity and the high time resolution of the terahertz spectroscopy and related detection methods provide a new visual angle for the research in the fields of biology, chemistry, physics and the like. However, water with a high content in a biological system has extremely strong absorption to the THz wave band, so that the research on the transmission spectrum of a liquid sample is relatively difficult. The method utilizes the property of the surfactant of the biological molecule, adopts a reverse micelle form, and detects the change of kinetic information of interface water molecules in a protein combination state through THz-TDS to represent the interaction between protein and phospholipid membrane.
At present, THz-TDS detection biomolecules mostly use solid tablets or hydrated biomolecules under certain humidity. This approach is far from the physiological existence of biomolecules and is often only suitable for the simplest single compound resolution. Taking a method for detecting the combination of phospholipid membrane and protein by THz-TDS as an example, the existing liposome system characterized by terahertz is obtained by adopting a vesicle encapsulation method: the method comprises evaporating phospholipid dissolved in organic solvent to form film, re-dissolving in dichloromethane, slowly injecting protein water solution as inner water phase, stirring, vortexing, and ultrasonically treating to obtain emulsion, evaporating at room temperature to remove organic phase, and adding water phase to obtain liposome. The phospholipid molecules are similar to the physiological state under the condition of using an extremely thin method for the formed liposome solution, but the water content of the system is extremely high, the absorption in THz is very strong, and the thickness of a sample is greatly limited; in addition, due to the water-soluble characteristics of proteins, free proteins exist outside vesicles, and it is difficult to ensure sufficient binding of proteins to phospholipid membranes.
In order to further expand the application of terahertz in the field of biological detection, researchers at home and abroad load a biomolecule detection object on a metamaterial structure in a spin coating mode based on an SRR (split ring resonator) type single-resonance metamaterial biosensor structure since 2010, and distinguish biological samples with close structures through the shift of different resonance peaks. The terahertz spectrum biological detection technology based on the metamaterial device has made a certain progress, but the terahertz spectrum biological detection technology still faces many difficulties, such as small measurement area, uneven sample and the like, which leads to inaccuracy of measurement results.
Disclosure of Invention
The invention aims to provide a method for detecting a liquid sample based on a terahertz technology, so as to solve the problem that in the prior art, the measurement result is inaccurate due to small measurement area and non-uniform sample in the terahertz spectrum biological detection technology.
In order to solve the technical problems, the invention adopts the following technical scheme:
the method for detecting the liquid sample based on the terahertz technology comprises the following steps: s1: providing a terahertz spectrum device, a container capable of containing a liquid sample, and a metamaterial sensor made of a terahertz metamaterial, wherein a periodic structure with a filtering or resonance function in a terahertz frequency band is carved on the metamaterial sensor, and the metamaterial sensor is arranged by clinging to the inner wall of one side of the container through one surface opposite to the periodic structure; s2: preparing a biomolecule to be detected into reverse micelle emulsion, and adding the reverse micelle emulsion into the container, wherein the reverse micelle emulsion is an emulsion with a continuous phase as an oil phase, a dispersed phase as a water phase and a phospholipid molecule as a surfactant, the phospholipid molecule exists in a reverse micelle form, the biomolecule to be detected is wrapped in the dispersed phase, and the reverse micelle emulsion and the metamaterial sensor form a whole body to be detected; and S3: and a terahertz spectrum device is used for obtaining the spectrum information of the total body to be detected in a terahertz wave band so as to realize the detection of the liquid sample.
According to a preferred embodiment of the present invention, the biomolecule to be detected in step S2 includes proteins, polypeptides, amino acids, phospholipids, nucleic acids, etc., wherein the proteins include water-soluble proteins, membrane-embedded proteins, membrane-bound proteins, etc.
According to a preferred embodiment of the present invention, in step S3, the quantitative determination of trace biomolecules, the characterization of binding and conformation changes of biomolecules, and the kinetic analysis of biochemical reactions can be achieved by using the terahertz spectroscopy apparatus to obtain the spectral information of the population to be measured in the terahertz waveband.
According to a preferred embodiment of the present invention, the spectral information of the terahertz waveband obtained in step S3 includes kinetic information of water molecules at the phospholipid membrane interface and low-frequency vibration information of the biomolecule to be detected in the terahertz waveband.
According to a preferred embodiment of the present invention, the terahertz spectrum device used in step S3 can generate and detect 0.1THz-10THz, i.e., the absorption and dispersion of the electromagnetic wave in the terahertz band by the sample.
According to a preferred embodiment of the present invention, in step S1, the periodic structure on the metamaterial sensor made of terahertz metamaterial includes: i-like shape, single bar shape, double bar shape, bull's eye shape, and the like.
According to a preferred embodiment of the present invention, in step S2, different single phospholipid molecules or a mixture of phospholipid molecules can be selected as the substance forming the reverse micelle according to the properties of the biomolecule to be detected; according to the property of the biomolecule to be detected, biomolecule solutions with different concentrations can be prepared to be used as the disperse phase of the reverse micelle emulsion; according to the property of the biomolecule to be detected, different ion solutions can be prepared to be used as buffer solutions to prepare biomolecule solutions of the reverse micelle emulsion disperse phase.
A variety of phospholipid molecules can be selected in two cases: 1) when the detection target contains a difference in the action or content of the membrane for different phospholipid components; 2) when the biomolecule in the micelle to be detected needs a fixed ratio of a plurality of phospholipid molecules as a surfactant to prepare stable reverse micelle emulsion.
According to a preferred embodiment of the present invention, the preparation of the reverse micelle emulsion in the step S2 includes the steps of: 1) drying phospholipid dissolved in volatile organic solvent with nitrogen, and vacuumizing to remove solvent to form film-shaped phospholipid; 2) dissolving film-shaped phospholipid in a continuous phase solvent; 3) dissolving the biomolecule to be detected into corresponding dispersed phase buffer solution to prepare uniform biomolecule solution to be detected; and 4) adding the biomolecule solution to be detected into the continuous phase solution dissolved with the phospholipid, and performing vortex and ultrasonic treatment to prepare the stable reverse micelle emulsion.
According to a preferred embodiment of the present invention, the container capable of containing the liquid sample in step S1 is a sample cell which has good permeability in the terahertz band and can serve to contain the liquid.
According to the method provided by the invention, the whole to be detected in the detection is positioned in a container which simultaneously contains the fluid and fixes the terahertz metamaterial, and the fluid is positioned on the surface of the terahertz metamaterial sensor.
According to the invention, the metamaterial sensor made of the terahertz metamaterial is a metamaterial sensor with specific resonant frequency in a terahertz frequency band, namely a periodic structure with a filtering/resonance function in the terahertz frequency band, the filtering property of the metamaterial sensor is determined by an artificial structure and is not mainly determined by the intrinsic property of a forming material, and the unit structure is equivalent to one tenth of the working wavelength. The metamaterial sensor based on the sub-wavelength metal strip array structure can realize different filtering functions of the sensor in a terahertz wave band, such as a single band, a double band and a broadband by changing the shape of a unit structure, and the unit structure is diversified in distribution form corresponding to different filtering functions.
It should be understood that metamaterial sensors of different unit structures can be selected to achieve better effects according to the required filtering function. For example, according to the preferred embodiment of the present invention, the h-shaped metamaterial sensor and the reverse micelle can be used to detect the conformation of the coagulation factor VIII protein, the bi-strip metamaterial sensor and the reverse micelle can be used to detect the conformation of the milk lectin protein, the h-shaped metamaterial sensor and the reverse micelle can be used to detect the binding of the coagulation factor VIII protein to the phospholipid membrane protein, the bull's eye metamaterial sensor and the reverse micelle can be used to detect the binding of the metal ions to the phospholipid membrane, the bi-strip metamaterial sensor and the reverse micelle can be used to detect the binding of the milk lectin to the phospholipid membrane, and the single strip metamaterial sensor and the reverse micelle can be used to detect the accumulation of the trace amyloid protein.
The invention creatively introduces the terahertz metamaterial and the reverse micelle for the first time, emphatically solves the defect of strong absorptivity of water with higher content in a biological system to a terahertz waveband, utilizes a terahertz spectrum device, provides a method capable of detecting a biological sample in solution at high sensitivity, and detects biological molecules by combining the terahertz metamaterial and the reverse micelle, thereby obviously improving the detection sensitivity and the applicability.
As described in the background section, the problem of detecting the combination of phospholipid membrane and protein by adopting the THz-TDS method in the prior art is that free protein exists outside vesicles due to the water-soluble characteristic of the protein, so that the sufficient combination of the protein and the phospholipid membrane is difficult to ensure.
Compared with the prior art, the method has the following remarkable advantages that 1) the method can detect the property, interaction and quantitative detection of the biological molecules under the microenvironment and phospholipid self-assembly form close to physiological conditions, 2) the detection sample is in a reverse micelle form, and the detection of the combination of phospholipid and protein with different targets can be achieved by slightly adjusting the preparation method of the reverse micelle, 3) the method can detect various proteins capable of being combined with membranes, such as lactadherin and the like, 4) the method is suitable for the detection of trace biological molecules and the representation of specific chemical properties based on the combination of terahertz metamaterials and a reverse micelle system, 5) the method can not only analyze the absorption coefficient α but also analyze the refraction rate change, dielectric constant and the like without using the parameters of terahertz time domain spectrum, 6) the method can also select dispersed phase ion solution according to the properties and the like of the sample to be detected, and the purposes of detecting the combination of copper ions, the valence state, concentration, protein and membrane combination of lead compounds and high-point screening can be achieved theoretically, and the detection of lead compounds and the lead compounds can be further screened rapidly.
In summary, according to the method for detecting the liquid sample based on the terahertz technology provided by the invention, the terahertz spectrum device is utilized, and based on the combination of the terahertz metamaterial and the reverse micelle, not only is the rapid and real-time detection realized, but also the recurrence of the protein and cell membrane combination state and the improvement of the sensitivity under the condition close to the real physiological condition are realized. In general, the present invention provides a method for sensitive detection of biomolecules based on terahertz spectroscopy.
Drawings
FIG. 1 is a schematic flow diagram of the process of the present invention;
FIG. 2 is a schematic diagram of a generic reverse micelle employed in the process of the present invention;
FIG. 3 is a schematic diagram of a reverse micelle sample for detection of the conformation of factor VIII protein according to a preferred embodiment of the present invention;
FIG. 4 is a schematic diagram of a terahertz time-domain spectroscopy device according to an embodiment of the present patent;
FIG. 5 is a microscopic image of an I-shaped metamaterial sensor according to an embodiment of the present invention;
FIG. 6 is a sample cell design incorporating metamaterials according to an embodiment of the present invention;
FIG. 7 is a schematic diagram of a reverse micelle sample for detecting the conformation of a milk lectin protein, according to a preferred embodiment of the present invention;
FIG. 8 is a schematic diagram of a dual strip metamaterial sensor in accordance with an embodiment of the present invention;
FIG. 9 is a schematic representation of a reverse micelle sample for detecting binding of factor VIII protein to a membrane protein according to a preferred embodiment of the present invention;
FIG. 10 is a schematic representation of a reverse micelle sample for detecting the binding of a milk lectin protein to a phospholipid membrane, according to a preferred embodiment of the present invention;
FIG. 11 is a sample terahertz absorption spectrum for detecting binding of milk lectin protein to phospholipid membrane, according to a preferred embodiment of the present invention;
FIG. 12 is a schematic diagram of a single strip metamaterial sensor in accordance with an embodiment of the present invention;
FIG. 13 is a sample terahertz absorption spectrum for detecting trace amyloid accumulations according to a preferred embodiment of the present invention;
FIG. 14 is a schematic view of a bullseye metamaterial sensor in accordance with an embodiment of the present invention;
wherein:
1-surfactant, 2-disperse phase, 3-continuous phase, 4-coagulation octafactor, 5-phosphate buffered saline solution, 6-quartz cuvette, 7-femtosecond laser, 8-spectroscopic sheet, 9-pump beam, 10-probe beam, 11-emitter, 12-time delay device, 13-receiver, 14-lock phase amplifier, 15-1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine, 16-sample cell, 17-reverse micelle emulsion, 18-metamaterial sensor, 19-lactadherin, 20-lipid anchored membrane protein, 21-1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine, 22, 23, 24. 25-the first, second, third, and fourth samples prepared in example 5, 26, 27, and 28-the three reverse micelle emulsions prepared in example 6.
Detailed Description
The present invention will be further described with reference to the following specific examples. It should be understood that the following examples are illustrative only and are not intended to limit the scope of the present invention.
According to the invention, a method for detecting a liquid sample based on a terahertz technology is provided, which comprises the following steps: obtaining spectrum information of a total body to be detected in a terahertz wave band by using a terahertz time-domain spectrum; detecting that the sample is in a sample container to be detected containing fluid; the inner wall of the container is of a single-resonance metamaterial biosensor structure; the sample is reverse micelle emulsion with continuous phase as oil phase, dispersed phase as water phase and phospholipid molecule as surfactant; wherein the phospholipids are present in the sample in the form of reversed micelles and the proteins are in the aqueous phase.
The flow of the method is shown in figure 1. Reverse micelle emulsion samples were first designed based on the identified examples of the binding of the exploratory protein to the membrane. The reverse micelle emulsion in the method is prepared by the following steps: drying phospholipid dissolved in volatile organic solvent with nitrogen, and vacuumizing to remove the solvent to form a film; dissolving film-shaped phospholipid in a continuous phase solvent; dissolving the protein into corresponding dispersed phase buffer solution; adding the protein solution into the continuous phase solution dissolved with the phospholipid, and preparing the stable reverse micelle emulsion by a vortex and ultrasonic method. The general reverse micelle form is shown in fig. 2 and comprises a surfactant 1, a dispersed phase 2 and a continuous phase 3. The single resonance metamaterial biosensor in the method is customized according to the dielectric properties of a biological sample and micelle and is tightly attached to the inner wall of a to-be-detected sample container containing fluid. The next process is to obtain the terahertz waveband optical property of the reverse micelle emulsion through terahertz time-domain spectroscopy. The terahertz time-domain spectrum device used by the method can generate and detect 0.1THz-10THz, namely the absorption and dispersion of the electromagnetic wave of the terahertz waveband through a sample. Information carried by a terahertz spectrum obtained by the electromagnetic wave of the terahertz waveband through a sample comprises kinetic information of interface water molecules and low-frequency vibration information of biomolecules in the terahertz waveband, and the information is further highlighted by the filtering action of the metamaterial biosensor. The next process is to analyze the detection of the sample on the obtained optical properties of the terahertz waveband to obtain the property information of the biomolecules under the condition close to the physiological condition.
Example 1 detection of factor VIII protein conformation in combination with an I-like metamaterial sensor and reverse micelles
A preferred embodiment of the method according to the invention for detecting properties of biomolecules is the detection of the overall conformational change of the coagulation factor VIII protein. In the present example, the sample is shown in fig. 3, the dispersed phase is formed by dissolving factor octa-coagulation 4 in phosphate buffered saline 5 with pH 7.3, the surfactant is 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15, and in order to examine the influence of the overall conformation of the calcium ion factor octa-coagulation protein, the control group is set to additionally contain 2mM CaCl2. Specifically, the test sample was prepared as follows: drying 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15 dissolved in trichloromethane by using nitrogen, vacuumizing and removing a solvent to form a film; dissolving the film-shaped phospholipid in hexadecane; dissolving coagulation factor VIII 4 in phosphate buffered saline 5 with pH 7.3 to obtain a uniform protein solution; adding the coagulation factor VIII protein solution into the continuous phase solution dissolved with phospholipid, and preparing the stable reverse micelle emulsion shown in figure 3 by a vortex and ultrasonic method.
In the present embodiment, a sample was tested using a terahertz time-domain spectroscopy apparatus as shown in fig. 4. The prepared reverse micelle emulsion was added to a 0.5mm quartz cuvette 6. The terahertz pulse generated by the femtosecond laser 7 is divided into a pumping beam 9 and a detection beam 10 by a beam splitter 8, and the light path of the detection beam 10 passes through the quartz cuvette 6 after passing through a transmitter 11. The probe beam 10 is passed through a time delay device 12 to adjust the time delay between the pump pulse and the probe pulse, and the signal is received by a receiver 13, passed through a lock-in amplifier 14, and the phase and amplitude of the signal at a certain frequency are measured. Finally, the whole time domain waveform of the terahertz pulse can be detected, and the frequency domain spectrum of the detected sample can be obtained through Fourier transform, so that the absorption coefficient, the refractive index, the transmissivity and other optical parameters of the sample can be obtained.
As shown in fig. 5, the h-shaped metamaterial sensor used in this embodiment is integrally made of a terahertz metamaterial, a plurality of h-shaped structures are carved on the h-shaped metamaterial, the size of each unit structure is one tenth of the working wavelength, and when the metamaterial sensor is perpendicular to an incident wave, a resonance peak with a strength of about 18dB exists at a position of 1.23 THz.
The fabrication of such i-shaped metamaterial sensors belongs to the prior art, and the fabrication method according to a preferred embodiment is provided herein by way of example only and not by way of limitation as follows: 1) and manufacturing a mask. Drawing a structure diagram of the surface of the device by using LEdit90 drawing software, drawing a structure to be etched in the software, and manufacturing a mask plate and a 5-inch chromium plate by adopting an electron beam exposure method according to the diagram. On the mask plate, the pattern to be remained is shaded, and the part to be etched is still transparent. 2) And (4) cleaning the material. And (4) putting the mask and the quartz plate into sulfuric acid and hydrogen peroxide cleaning solution for cleaning. 3) And (6) coating photoresist. Photoresists are typically used as media for transferring optical, electron beam, or ion beam lithographic patterns. The photoresist can be divided into positive photoresist and negative photoresist, and the positive photoresist is used in the process, namely the photoresist is dissolved in a developing solution after exposure, and the unexposed part is not dissolved. And (2) gluing the quartz wafer by using a spin coater, firstly spin-coating a layer of adhesion promoter on the surface of the wafer to increase the viscosity between the wafer and the photoresist, then gluing, and controlling the gluing thickness by the rotating speed and the spinning time of the spin coater, wherein the coating thickness is 5 mu m. And finally, putting the sample into a dryer for drying, and evaporating the solvent in the photoresist to control the sensitivity of the photoresist and release stress. 4) And (6) ultraviolet exposure. And pressing the quartz wafer coated with the photoresist under a mask plate for ultraviolet exposure, wherein the exposure time depends on the sensitivity and the thickness of the photoresist and the power of ultraviolet light. And then putting the sample into a dryer again for drying and shaping so that the exposed photoresist is subjected to chemical reaction and the pattern is more uniform. 5) And (6) developing. The sample is placed in a developing solution for development, and the exposed photoresist is dissolved in the developing solution, while the unexposed portion remains coated on the surface of the sample. And baking the sample in a dryer again to further shape the pattern, and evaporating the residual developing solution. 6) And (5) plating metal. Because gold plating directly on the glass substrate is easy to detach, a layer of 10nm chromium is plated on the substrate as an adhesion layer, and then gold plating is carried out, wherein the thickness of a gold film is about 500 mu m. 7) And removing the photoresist and cleaning. The photoresist removing gas is oxygen, and is ionized under high voltage to generate oxygen ions, so that the photoresist is oxidized into volatilizable gas which is finally pumped away by a suction pump. 8) And (6) scribing. Taking unimodal metamaterial samples and bimodal metamaterial samples as examples, a plurality of samples can be designed on a quartz wafer, and after the processing is finished, the samples need to be scribed by laser. And finishing the manufacture of the metamaterial sensor carved with a plurality of periodic structures.
According to the preferred embodiment, the sample cell 16 for terahertz spectrum testing is shown in fig. 6, the metamaterial sensor 18 is arranged by closely attaching a surface opposite to the h-shaped structure to the inner wall of one side of the sample cell 16, the surface with the h-shaped structure faces the fluid, the fluid in the sample cell 16 is reverse micelle emulsion 17, and the reverse micelle emulsion 17 is located on the surface of the metamaterial sensor 18 during measurement. In conclusion, the terahertz time-domain spectroscopy measures the terahertz waveband spectrum information of the quartz cuvette 6 containing the sample. In the embodiment, the characteristics and the sensitivity of the reverse micelle system combined with the metamaterial sensor in the detection of the biomolecules are obviously improved.
Since the phospholipid used for forming the micelle in this embodiment is 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 that does not specifically bind to factor viii 4, the micelle only provides a micro-nano-scale microenvironment of the aqueous solution and does not participate in the conformational change of the protein. Coagulation factor VIII 4 protein binding ligands such as copper ions, calcium ions and the like are added into the disperse phase aqueous solution, so that the coagulation factor VIII 4 reverse micelles under different conditions are obtained. Due to the ligand binding effect, the addition of different ligand ions or no ligand ions can cause the overall conformation change of the protein and the change of the solvent and the surface of the corresponding protein, so that the protein can be distinguished by terahertz. The overall conformational change of the protein is captured by the terahertz time-domain spectroscopy, and because the crystal structure of the coagulation factor VIII 4 protein is known, information related to the overall hydration interface of the coagulation factor VIII protein, such as the change of the hydration degree and the surface hydrophilicity and hydrophobicity along with the overall conformational change of the protein in an aqueous solution, can be obtained by comparing the terahertz time-domain spectroscopy information obtained by the method with a computational chemical simulation result.
Example 2 detection of milk agglutinin protein conformation by combining double strip metamaterial sensor and reverse micelle
A preferred embodiment of the method according to the invention for detecting properties of biomolecules is the detection of the overall conformational change of the milk lectin protein. In the present example, the test sample is shown in fig. 7, the dispersed phase is composed of lactadherin 19 dissolved in phosphate buffered saline 5 with pH 7.3, the surfactant is 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15, and in order to examine the influence of the overall conformation of the calcium ion coagulation factor viii protein, the control group is set to additionally contain 2mM CaCl2. The test sample was prepared as follows: will dissolve in trichloro benzeneDrying the 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15 of methane by using nitrogen, vacuumizing and removing the solvent to form a film; dissolving the film-shaped phospholipid in hexadecane; dissolving lactadherin 19 in phosphate buffered saline 5 with pH 7.3 to obtain uniform protein solution; adding the coagulation factor VIII protein solution into the continuous phase solution dissolved with phospholipid, and preparing the stable reverse micelle emulsion by a vortex and ultrasonic method. A typical terahertz spectroscopy apparatus is shown in fig. 4. The sample cell structure is shown in fig. 6, the only difference is the difference of the periodic structure on the metamaterial sensor 18, the dual-strip metamaterial sensor is shown in fig. 8, the metamaterial sensor 18 is arranged by the surface opposite to the dual-strip structure and clings to the inner wall of one side of the sample cell 16, and the fluid in the sample cell is reverse micelle emulsion 17 loaded with coagulation factor eight. In the embodiment, the characteristics and the sensitivity of the reverse micelle system combined with the metamaterial sensor in the detection of the biomolecules are obviously improved.
Since the phospholipid used to form the micelle in this example is 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 that does not specifically bind to lactadherin 19, the micelle only provides a micro-nano-scale aqueous solution microenvironment and does not participate in the conformational change of the protein. By changing the microenvironment of the dispersed phase aqueous solution, such as pH value, ion concentration and the like, the whole conformational change of the lactadherin 19 protein and the change of the solvent and the surface of the corresponding protein can be caused, so that the lactadherin can be distinguished by terahertz. Since the crystal structure of lactadherin has not been completely resolved, the ligand binding effect and the overall conformation cannot be obtained with a more accurate result in the simulation calculation. However, the method provided by the invention can be used for summarizing the conformation-related binding ligand or weak bond action of the protein by detecting the overall conformation change of the protein, so as to obtain the property information of related biomolecules in a near-physiological solution state.
Example 3 detection of binding of factor VIII protein to phospholipid Membrane protein based on an I-shaped metamaterial-like sensor and reverse micelle
A preferred embodiment for detecting the biomolecular interactions according to the method of the present invention is the detection of the binding of factor VIII protein 4 to the phospholipid membrane of 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 loaded with lipid-anchored membrane protein 20. The prepared reverse micelle sample is shown in fig. 9. Wherein, the disperse phase is formed by dissolving coagulation eight factor protein 4 in phosphate buffered saline 5 with pH 7.3; the phospholipid is composed of 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 and lipid-anchored membrane protein 20 in a mass ratio of 20:1, and is used for detecting the binding condition of different contents of lipid-anchored membrane protein 20. The preparation method of the sample is as follows: drying 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15 dissolved in chloroform by using nitrogen, adding lipid anchoring membrane protein 20, and vacuumizing to remove the solvent to form a film; dissolving the film-shaped phospholipid in hexadecane; lipoanchored membrane protein 20 was dissolved in phosphate buffered saline at pH 7.3; adding the protein solution into the continuous phase solution dissolved with the phospholipid, and preparing the stable reverse micelle emulsion by a vortex and ultrasonic method. A typical terahertz detection device is shown in fig. 4, a sample cell structure is shown in fig. 6, one surface of an i-shaped terahertz metamaterial-like sensor 18 opposite to the i-shaped structure is placed close to the inner wall of a sample cell 16, and a characteristic absorption peak is present in pure hexadecane at 1.3 THz. To detect binding of factor viii to membrane proteins, paired samples containing and not containing lipoanchored membrane protein 20 were prepared and compared by terahertz spectroscopy control. The terahertz waveband optical property of the reverse micelle emulsion is obtained through terahertz time-domain spectroscopy, and the responsiveness of the lipid-anchored membrane protein 20 to the combination of coagulation factor VIII 4 can be analyzed and obtained.
Example 4 detection of milk agglutinin binding to phospholipid Membrane based on double strip-shaped metamaterial sensor and reverse micelle
One preferred embodiment for detecting binding of biomolecules according to the method of the present invention is the detection of binding of milk agglutinin to phospholipid membranes. As shown in FIG. 10, the test sample had a dispersion phase comprising a solution of lactadherin 19, and a control group containing 2mM CaCl was set for examining the effect of calcium ion on binding of lactadherin 20 to the membrane2(ii) a Phospholipid 2 is prepared by mixing 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15 and 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine 21 according to a certain proportion, and is used for detecting the different content of 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine 21 to the milk agglutinin19. The test sample was prepared as follows: mixing 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 19 and 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine 21 dissolved in chloroform with phospholipid, drying by using nitrogen, vacuumizing, and removing the solvent to form a film; dissolving the film-shaped phospholipid in hexadecane; dissolving the milk agglutinin 19 into the buffer solution; adding the protein solution into the continuous phase solution dissolved with the phospholipid, and preparing the stable reverse micelle emulsion by a vortex and ultrasonic method. The typical terahertz spectrum and the application method of the double-strip metamaterial are the same as those in the embodiment 2.
The sample absorption spectrum of this example is shown in FIG. 11. Under the THz wave band of 0.2-1.4, the mixed phospholipid 2 with the mass fraction of 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 15 of 95 percent and the mass fraction of 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine 21 of 5 percent is shown, and CaCl is not contained in the dispersed phase 32The first sample 22, 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine 19 mass fraction is 75% and the 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine 21 mass fraction is 25%, the mixed phospholipid 2 is formed, and the dispersed phase 3 does not contain CaCl2The second sample 23, 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 mass fraction of 95% and 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine 21 mass fraction of 5% comprised mixed phospholipid 2, the dispersed phase 3 contained 2mM CaCl2The third sample 24, 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine 15 mass fraction of 75% and 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine 21 mass fraction of 25% comprised mixed phospholipid 2, dispersed phase 3 containing 2mM CaCl2The absorption coefficients of the fourth sample 25 and the above four groups of samples are shown as α. it can be seen from the graph that the absorption coefficients α of the first sample 22 and the third sample 24 and the second sample 23 and the fourth sample 25 are substantially consistent in the frequency range of 0.2-1.4THz, which indicates that calcium ions do not affect the selective binding of the lectin 19 to the phospholipid membrane, while the absorption coefficients α of the first sample 22 and the second sample 23 and the third sample 24 and the fourth sample 25 are greatly different, which indicates that the mass fraction of the 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine 21 affects the binding of the lectin 19 to the phospholipid membrane.
Example 5 detection of Trace amyloid aggregates based on Single Bar metamaterial Sensors and reverse micelles
A preferred embodiment of the method according to the invention for detecting trace amounts of biomolecules is the detection of trace amounts of amyloid aggregates. In the embodiment, the characteristics and the sensitivity of the reverse micelle system combined with the metamaterial sensor in the detection of the biomolecules are obviously improved. Based on micelle stability this example selected 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine: 1, 2-dioleoyl-sn-glycero-3-phosphatidylethanolamine is 3: 1 as a surfactant to prepare the reversed micelle loaded with amyloid protein aggregates. We first analyzed the reverse micelle sample cell in the basic system. We chose to prepare 5.0. mu.M of monomeric amyloid acculation solution and then dilute it to obtain different concentrations of aggregation solution. In combination with the fact that low concentrations of accumulated solution are more significant for the effect of reverse micelles. This example probes terahertz spectra obtained by loading several groups of solutions of different concentrations in reversed micelles diluted with an amyloid accumulating solution. After the amyloid accumulation solution is added, the absorption characteristics of the reverse micelle sample are changed, compared with 5 accumulation solutions with different concentrations between 0.01 mu M and 1.0 mu M, the absorption characteristics are basically not influenced when the concentration of the accumulation solution is very low (0.01 mu M), and the terahertz absorption coefficient is obviously reduced when the concentration reaches 0.1 mu M and is changed more obviously along with the increase of the concentration.
Subsequently, based on characterization control of a single-peak terahertz frequency, we selected a single-strip metamaterial resonance structure, as shown in fig. 12, to be combined with an inverse micelle system to improve the detection sensitivity for amyloid. Sample absorption spectrum of this example referring to FIG. 13, when the metamaterial sensor is placed in the sample cell, the absorption coefficient 26 of the reversed micelle has an intensity of about 71.2cm at 0.72THz-1Has an absorption coefficient 27 of reversed micelles of concentration 0.001. mu. M A β 42, and has an intensity of about 66.3cm at 0.74THz-1Has an absorption coefficient 28 of reversed micelles of concentration 0.01. mu. M A β 42, and has an intensity of about 62.5cm at 0.76THz-1The resonance peak of (1).In conclusion, by introducing the terahertz metamaterial, the detection limit of the method on trace amyloid accumulation is reduced by at least one order of magnitude compared with the detection limit without introducing the metamaterial.
Example 6 combination of bovine-eye metamaterial sensor and reverse micelle detection of binding of metal ions to phospholipid Membrane
One preferred embodiment for detecting biomolecules according to the method of the invention is the detection of binding of metal ions to phospholipid membranes. In this embodiment, phospholipid molecules constituting the reverse micelle are the detection objects, and the pre-detection is performed to obtain a better discrimination for water at the phospholipid membrane interface by using a bullseye-shaped metamaterial sensor, the bullseye-shaped metamaterial sensor used in this embodiment is shown in fig. 14, the used test sample is a water layer to prepare a reverse micelle sample wrapping copper ions, and the phosphate cluster formed by the polar groups at the head of the phospholipid molecules in the reverse micelle form is characterized by using the amphipathy of the phospholipid molecules and the form of the reverse micelle and detecting the change of the kinetic information of the interface water molecules in the binding state through terahertz time-domain spectroscopy. Phosphatidylcholine phospholipid is the most ubiquitous phospholipid in biological systems, and has no specific binding effect with copper ions in previous researches, so that the effect of copper ions on different phosphate clusters is represented by terahertz by respectively preparing 1, 2-dioleoyl-sn-glycerol-3-phosphatidylcholine reverse micelles and 1, 2-dioleoyl-sn-glycerol-3-phosphatidylglycerol reverse micelles wrapping copper ion solutions. The influence of copper ions on the polarization and spatial arrangement of the original hydration layer of phospholipid molecules is detected through terahertz time-domain spectroscopy. The phospholipid molecular film layer with a large surface and ordered spatial arrangement in the system is exposed in the copper ion solution, so that the detection sensitivity is improved, and the absorption of bulk water to the terahertz wave band is reduced to the maximum extent. The terahertz detection structure shows that the absorption coefficient of the DOPG reverse micelle gradually becomes stronger and the increment of the absorption coefficient also gradually increases with the concentration of copper ions from 0.2mM to 20 mM. An increase in the copper ion concentration causes a decrease in the water content in the reverse micelle system, and a decrease in the water content causes a decrease in the value of the absorption coefficient of the reverse micelle. In order to better analyze the effect of the phosphate clusters and the copper ions, as a control, the present example also supplements the reverse micelle test in which the dispersed phase is an aqueous solution of magnesium ions. In combination with the above detection results, the present embodiment can distinguish the specific binding effect between different trace metal ions and different phospholipid cluster polar heads. Considering that the distribution of bulk water and interfacial water in the system can be influenced while the concentration of copper ions is increased, the interfacial binding effect of different phospholipid clusters can be deduced, so that the purpose of detecting the binding effect of biomolecules is achieved.
The above embodiment is an implementation demonstration of reverse micelle-based terahertz biological detection, and aims to illustrate that a phospholipid reverse micelle-based system can have a wide range of applications in biomolecule detection and property characterization. By combining the terahertz metamaterial and the reverse micelle, the detection sensitivity can be further improved, and the specificity of the terahertz spectrum is highlighted. The following examples are intended to focus on the design of the probing system and the test method.
Example 7 measurement of phospholipase Activity
A preferred example of the use of the method according to the invention for the quantitative determination of the progress of a biomolecular reaction is the measurement of phospholipase activity. This example is a measurement of the catalytic activity of phospholipase on 1, 2-dioleoyl-sn-glycero-3-phosphatidylserine. The test sample in the reverse micelle form included a dispersed phase, a continuous phase. In a reverse micelle system, phospholipase active sites are combined to catalyze 1, 2-dioleoyl-sn-glycerol-3-phosphatidylserine to react and decompose, a reaction product causes the reverse micelle to change, and the change can be detected by terahertz time-domain spectroscopy through the change of a biomolecule hydration interface at the initial stage of reaction. The time-dependent micelle system change process can be obtained through continuous terahertz spectrum testing, so that a dynamic curve of phospholipase catalytic reaction can be quantitatively drawn.
The terahertz metamaterial sensor is added into a reverse micelle system, so that the detection sensitivity of the terahertz metamaterial sensor to changes in the reaction process of the system can be further increased, and the purpose of measuring micro-reaction is achieved. By the method of this embodiment, the reaction constants of different phospholipases can be measured, and the conditions under which the different phospholipases catalyze a reaction can be probed. In particular, when the method provided by the present invention is used to measure the progress of a biomolecular reaction, it is not a requirement for accurate description of the state and conformation of the biomolecules in the system. Because of the rate of change information given based on continuum, it has been possible to infer the reaction mechanism, leading to valuable relevant chemical reaction kinetics information such as the number of reaction stages, reaction rate constants, etc.
Example 8 probing of nucleic acid base sequences
Another preferred embodiment of the method according to the invention for detecting biomolecules is the detection of nucleic acid sequences. In this example, reverse micelle samples of deoxyribonucleic acids of known sequence were prepared: the disperse phase is pure water solution of deoxyribonucleic acid with single sequence type, and the phosphatidylcholine forming the reverse micelle does not react with the nucleic acid. The prepared reverse micelle samples are combined with terahertz metamaterial sensors such as a bullseye-shaped metamaterial sensor, an I-shaped metamaterial-like sensor and a single-strip metamaterial sensor one by one, and terahertz absorption spectra under multiple sensors of the deoxyribonucleic acid reverse micelle of the same sequence are obtained. And directly inputting the obtained plurality of terahertz absorption spectra as characteristic parameters, and modeling and training a machine learning model for predicting the deoxyribonucleic acid sequence according to the reverse micelle terahertz absorption spectra under multiple conditions. The prediction model obtained based on the training set can be used for detecting the base sequence of the deoxyribonucleic acid at the position: preparing deoxyribonucleic acid with an unknown sequence into a reverse micelle sample, combining terahertz metamaterial sensors such as a bullseye-shaped metamaterial resonant structure, an I-shaped metamaterial sensor and a single-strip-shaped metamaterial sensor one by one, wherein different terahertz metamaterial sensors can obtain sample terahertz spectrograms under different characteristic peak types, inputting a plurality of obtained terahertz absorption spectrums serving as characteristic parameters, obtaining terahertz absorption spectrums under a plurality of sensors of the deoxyribonucleic acid reverse micelle with the same sequence, inputting the terahertz absorption spectrums into a machine learning prediction model, and predicting a base sequence. In the method, the sample in the reverse micelle mode highlights the terahertz frequency band spectrum information of the biological molecules to the maximum extent, and the combination of the sample and the terahertz metamaterial enables the same sample to be detected into a plurality of characteristic absorption spectrums. The detection accuracy of the method can be gradually improved along with the improvement of the artificial intelligence and terahertz spectrum device technology, and the method finally has the capabilities of instantaneity, low cost and high accuracy.
The above embodiments are merely preferred embodiments of the present invention, which are not intended to limit the scope of the present invention, and various changes may be made in the above embodiments of the present invention. All simple and equivalent changes and modifications made according to the claims and the content of the specification of the present application fall within the scope of the claims of the present patent application. The invention has not been described in detail in order to avoid obscuring the invention.
Claims (10)
1. A method for detecting a liquid sample based on a terahertz technology is characterized by comprising the following steps:
s1: providing a terahertz spectrum device, a container capable of containing a liquid sample, and a metamaterial sensor made of a terahertz metamaterial, wherein a periodic structure with a filtering or resonance function in a terahertz frequency band is carved on one surface of the metamaterial sensor, and the metamaterial sensor is arranged by clinging to the inner wall of one side of the container through the surface opposite to the periodic structure;
s2: preparing a biomolecule to be detected into reverse micelle emulsion, and adding the reverse micelle emulsion into the container, wherein the reverse micelle emulsion is an emulsion with a continuous phase as an oil phase, a dispersed phase as a water phase and a phospholipid molecule as a surfactant, the phospholipid molecule exists in a reverse micelle form, the biomolecule to be detected is wrapped in the dispersed phase, and the reverse micelle emulsion and the metamaterial sensor form a whole body to be detected; and
s3: and a terahertz spectrum device is used for obtaining the spectrum information of the total body to be detected in a terahertz wave band so as to realize the detection of the liquid sample.
2. The method according to claim 1, wherein the test biomolecules in step S2 comprise proteins, polypeptides, amino acids, phospholipids and nucleic acids, wherein the proteins comprise water-soluble proteins, membrane-embedded proteins and membrane-bound proteins.
3. The method according to claim 1, wherein the step S3 of obtaining the spectrum information of the population to be measured in the terahertz waveband by using the terahertz spectrum device can realize the quantification of trace biomolecules, the characterization of the binding and conformation change of the biomolecules, and the kinetic analysis of biochemical reaction.
4. The method according to claim 1, wherein the spectral information of the terahertz waveband obtained in step S3 includes kinetic information of water molecules at the phospholipid membrane interface and low-frequency vibration information of the biomolecule to be detected in the terahertz waveband.
5. The method as claimed in claim 1, wherein the terahertz spectrum device used in step S3 is capable of generating and detecting 0.1THz-10THz, i.e. absorption and dispersion of electromagnetic waves in the terahertz band through the sample.
6. The method according to claim 1, wherein in step S1, the periodic structure on the metamaterial sensor made of terahertz metamaterial comprises: i-like shape, single bar shape, double bar shape and bull's eye shape.
7. The method according to claim 1, wherein, in step S2,
according to the property of the biomolecule to be detected, different single phospholipid molecules or mixed components of a plurality of phospholipid molecules can be selected as substances for forming reverse micelles;
according to the property of the biomolecule to be detected, biomolecule solutions with different concentrations can be prepared to be used as the disperse phase of the reverse micelle emulsion;
according to the property of the biomolecule to be detected, different ion solutions can be prepared to be used as buffer solutions to prepare biomolecule solutions of the reverse micelle emulsion disperse phase.
8. The method of claim 7, wherein the plurality of phospholipid molecules are selected in two cases: 1) when the detection target contains a difference in the action or content of the membrane for different phospholipid components; 2) when the biomolecule in the micelle to be detected needs a fixed ratio of a plurality of phospholipid molecules as a surfactant to prepare stable reverse micelle emulsion.
9. The method of claim 1, wherein the preparing of the reverse micelle emulsion in the step S2 comprises the steps of:
1) drying phospholipid dissolved in volatile organic solvent with nitrogen, and vacuumizing to remove solvent to form film-shaped phospholipid;
2) dissolving film-shaped phospholipid in a continuous phase solvent;
3) dissolving the biomolecule to be detected into corresponding dispersed phase buffer solution to prepare uniform biomolecule solution to be detected;
4) adding the biomolecule solution to be detected into the continuous phase solution dissolved with the phospholipid, and carrying out vortex and ultrasonic treatment to prepare the stable reverse micelle emulsion.
10. The method according to claim 1, wherein the container capable of containing the liquid sample in step S1 is a sample cell which is permeable in the terahertz wave band and can bear the function of containing liquid.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111811998A (en) * | 2020-09-01 | 2020-10-23 | 中国人民解放军国防科技大学 | Method for determining strongly-absorbable biological particle component under target waveband |
| CN111965133A (en) * | 2020-07-22 | 2020-11-20 | 中国南方电网有限责任公司电网技术研究中心 | Method for detecting water content and polarization form of micro-water-containing oil-immersed insulating paperboard |
| CN113138201A (en) * | 2021-03-24 | 2021-07-20 | 北京大学 | Metamaterial Internet of things system and method for wireless passive environment state detection |
| CN113686735A (en) * | 2021-09-16 | 2021-11-23 | 北京信息科技大学 | Method and device for measuring blood coagulation property |
| CN114088656A (en) * | 2020-07-31 | 2022-02-25 | 中国科学院上海高等研究院 | Terahertz spectrum substance identification method and system, storage medium and terminal |
| CN114279998A (en) * | 2021-12-29 | 2022-04-05 | 福州大学 | Liquid enhanced sensing system and method based on quasi-square-shaped terahertz metamaterial |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106198444A (en) * | 2016-04-19 | 2016-12-07 | 中国科学院上海应用物理研究所 | Microemulsion containing phospholipid and preparation method, application in the aqueous energy of research interface |
| US20170205403A1 (en) * | 2015-07-17 | 2017-07-20 | Zhejiang University | Biological sample signal amplification method using both terahertz metamaterials and gold nanoparticles |
| KR20180017735A (en) * | 2016-08-10 | 2018-02-21 | 아주대학교산학협력단 | Real time ion sensors using the same using terahertz metamaterials |
| CN110455743A (en) * | 2019-08-19 | 2019-11-15 | 中央民族大学 | Utilize the method for terahertz wave band Meta Materials sensor detection aflatoxin B1 and B2 |
| CN110632291A (en) * | 2019-09-26 | 2019-12-31 | 中国科学院半导体研究所 | Terahertz metamaterial biosensor and preparation method and detection method thereof |
| CN110658154A (en) * | 2019-09-29 | 2020-01-07 | 张阳 | Preparation and application of reproducible terahertz biological sample detection cell |
-
2020
- 2020-03-20 CN CN202010202734.3A patent/CN111307755A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170205403A1 (en) * | 2015-07-17 | 2017-07-20 | Zhejiang University | Biological sample signal amplification method using both terahertz metamaterials and gold nanoparticles |
| CN106198444A (en) * | 2016-04-19 | 2016-12-07 | 中国科学院上海应用物理研究所 | Microemulsion containing phospholipid and preparation method, application in the aqueous energy of research interface |
| KR20180017735A (en) * | 2016-08-10 | 2018-02-21 | 아주대학교산학협력단 | Real time ion sensors using the same using terahertz metamaterials |
| CN110455743A (en) * | 2019-08-19 | 2019-11-15 | 中央民族大学 | Utilize the method for terahertz wave band Meta Materials sensor detection aflatoxin B1 and B2 |
| CN110632291A (en) * | 2019-09-26 | 2019-12-31 | 中国科学院半导体研究所 | Terahertz metamaterial biosensor and preparation method and detection method thereof |
| CN110658154A (en) * | 2019-09-29 | 2020-01-07 | 张阳 | Preparation and application of reproducible terahertz biological sample detection cell |
Non-Patent Citations (6)
| Title |
|---|
| GIULIA FOLPINI ET AL.: "Water Librations in the Hydration Shell of Phospholipids", 《JOURNAL OF PHYSICAL CHEMISTRY LETTERS》 * |
| YANG JING ET AL.: "The terahertz dynamics interfaces to ion–lipid interaction confined in phospholipid reverse micelles", 《CHEMICAL COMMUNICATION》 * |
| 李明亮: "基于太赫兹时域光谱技术的生物分子鉴别及相互作用研究", 《中国博士学位论文全文数据库 基础科学辑》 * |
| 潘亚涛等: "基于太赫兹光谱技术的生物膜界面水研究", 《激光与光电子学进展》 * |
| 田震等: "太赫兹亚波长人工结构传感应用", 《物理》 * |
| 齐亮等: "食品中生物分子的太赫兹特性探测方法", 《包装与食品机械》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111965133A (en) * | 2020-07-22 | 2020-11-20 | 中国南方电网有限责任公司电网技术研究中心 | Method for detecting water content and polarization form of micro-water-containing oil-immersed insulating paperboard |
| CN114088656A (en) * | 2020-07-31 | 2022-02-25 | 中国科学院上海高等研究院 | Terahertz spectrum substance identification method and system, storage medium and terminal |
| CN111811998A (en) * | 2020-09-01 | 2020-10-23 | 中国人民解放军国防科技大学 | Method for determining strongly-absorbable biological particle component under target waveband |
| CN113138201A (en) * | 2021-03-24 | 2021-07-20 | 北京大学 | Metamaterial Internet of things system and method for wireless passive environment state detection |
| CN113138201B (en) * | 2021-03-24 | 2022-05-20 | 北京大学 | Metamaterial Internet of things system and method for wireless passive environment state detection |
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| CN114279998B (en) * | 2021-12-29 | 2024-01-09 | 福州大学 | Liquid enhancement sensing system and method based on quasi-Chinese character 'Hui' type terahertz metamaterial |
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