• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    The present invention discloses an environmentally responsive sensing device and its manufacturing methods and uses. The sensing device responsive to the environment comprises a patterned substrate, wherein at least two spaced-apart trenches are formed in the surface of the substrate, wherein the at least two trenches are filled with initiator-containing first polymers in order to provide a planar pattern as a detection area on the surface of the substrate form; and a plurality of second polymer graft chains and labeled macromolecules, the front ends of the second polymer graft chains being covalently bonded to active functional groups present on the surface of the first polymer and to the planar pattern, the rear ends of the second polymer graft chains being simultaneously bound by a nucleophilic substitution reaction are covalently bound to the labeled macromolecules, the labeled macromolecules being selected from a group consisting of block polymers, the block polymers having fluorescent groups and functional groups that can specifically bind to selected molecules, and where a number average molecular weight of the block polymers is 300 to 450. In the invention, the polymer graft chains can react to different solvents or to a particular molecule in order to achieve an automatic control function responsive to the environment.
    公开号:CH715741B1
    申请号:CH00731/20
    申请日:2018-07-26
    公开日:2021-10-29
    发明作者:Li Guangshuo；Wang Jie；Zhu Yixin；Jiang Yinglong；Yang Zonghui；Wang Min；Zheng Jingjing；Chen Gun
    申请人:Cqc Intime Testing Tech Co Ltd；
    IPC主号:
    专利说明:

    Field of invention
    The present invention relates to a sensing device and, more particularly, to an environmentally responsive sensing device and methods of manufacture and applications thereof.
    State of the art
    In the "development and performance optimization of materials that react to the environment" attempts to independently use materials that react to the environment in order to modify the substrates of conventional devices and to assemble them with them and thus control and adapt the surface properties of substrates To enable change of the environmental conditions. By detecting the substrate morphology or detecting the fluorescence signal, these changed physical or chemical quantities can be converted into identifiable signals in order to achieve the properties required for materials. Such materials can be used in the field of environmental monitoring and automation control. In conventional sensor materials, the conformational change of the materials is caused by the change in the environmental conditions. For example, this change can be caused by the isomerization of the functional groups of a material such as e.g. B. cis-trans isomerization and ionization of the chromophore, can be achieved, which leads to changes in the material properties.
    Atom Transfer Radical Polymerization (ATRP) is currently a very widespread controlled polymerization process (WANG J.-S., MATYJASZEWSKI K. Controlled / “living” radical polymerization. Atom transfer radical polymerization in the presence of transition-metal complexes [ J]. Journal of the American Chemical Society, 1995, 117 (20): 5614-5615.) and can be used extensively for the construction of composite blocks and for the production of functional polymer materials. However, there has been no report in the industry on the use of this method to manufacture an environment responsive sensing device.
    Object of the invention
    It is the primary object of the present invention to provide an environment responsive sensing device and its method of manufacture and applications by which the deficiencies of the prior art can be overcome.
    To achieve the above object, the following technical solutions are used in the present invention: According to the invention, a sensing device that is responsive to the environment is provided, which comprises: a patterned substrate, with at least two spaced-apart trenches in the surface of the substrate are formed, wherein the at least two trenches are filled with initiator-containing first polymers to form a planar pattern as a detection area on the surface of the substrate; and a plurality of second polymer graft chains and labeled macromolecules, the front ends of the second polymer graft chains being covalently bonded to active functional groups present on the surface of the first polymer and to the planar pattern, the rear ends of the second polymer graft chains being simultaneously bound by a nucleophilic substitution reaction are covalently bound to the labeled macromolecules, the labeled macromolecules are selected from a group consisting of block polymers, the block polymers fluorescent groups and functional groups that can specifically bind to selected small molecules, and wherein a number average molecular weight of Block polymers is 300 to 450; according to the invention, when the second polymer graft chains are added to a first solvent, the second polymer graft chains are in a stretched state, wherein when the second polymer graft chains are added to a second solvent, the second polymer graft chains are in a collapsed state, the length the second polymer graft chain in the stretched state is at least 6 to 10 times the length of this in the collapsed state; wherein, further, when the sensing area of the environmentally responsive sensing device is in contact with the first solvent, all of the plurality of second polymer grafts are stretched to form an upright bristle-like structure, wherein when the sensing area of the environmentally responsive sensing device is in contact with the second solvent is in contact, all of the multiple second polymer graft chains on the surface of the substrate have collapsed and there are clear boundaries between the spaced planar patterns. According to the invention there is provided a method of manufacturing the above-mentioned environment-responsive detection device comprising the following steps: (1) providing a patterned substrate, wherein at least two spaced-apart trenches are formed in the surface of the substrate; (2) Filling the precursor of the active functional group-containing first polymers into the trenches located on the surface of the substrate and then aligning and pressing a plane on the surface of the substrate at the polymerization temperature, the surface energy of the plane being higher than that of the substrate, to form a planar pattern as a detection area on the surface of the substrate; (3) covalently bonding the leading ends of the plurality of second polymer graft chains to the active functional groups on the surface of the first polymers by atom transfer radical polymerization; (4) covalently attaching the rear ends of the second polymer graft chains to the labeled macromolecules by a nucleophilic substitution reaction; wherein step (4) comprises: bringing a liquid phase reaction system containing the labeled macromolecules and a solvent into contact with the detection area and forming the second polymer graft chains in the stretched state and covalently bonding the rear ends of the second polymer graft chains through a nucleophilic substitution reaction the labeled macromolecules. According to the invention, a detection method is provided which is based on the above-mentioned detection device that reacts to the environment. The detection method includes: bringing the liquid sample to be tested, which contains a solvent and selected molecules, into contact with the detection area of the sensing device that is responsive to the environment, and then detecting the change in fluorescence intensity of the detection area to reflect that in the liquid to be tested Detect selected molecules in the sample.
    The advantageous effects of the present invention achieved in comparison with the prior art are described below: In the detection device according to the invention which reacts to the environment, the detection device can be precisely controlled by changing the solvent properties or specific substrates can be used for measuring the molecular properties of solutions be designed. By adapting the types of material at the rear ends of the polymer graft chains, the detection device can adapt to various environmental conditions, such as e.g. B. different reagent concentrations, different temperatures and different electric and magnetic fields, react in order to achieve an automatic control function that is responsive to the environment. In this way a new possibility for developing materials is provided. The invention has the prospect of wide application in the field of automatic surveillance, such as e.g. B. Monitoring of the steady state of colloidal particles, adsorption, liquid crystal displays, and water and gas pollutants, and is also an important fundamental unit of information acquisition for future data development.
    Brief description of the drawings
    For a better understanding of the technical solutions according to the exemplary embodiments of the invention, the drawings according to the exemplary embodiments of the invention are briefly described below. It will be understood that the drawings illustrate only a few exemplary embodiments of the present invention and that one skilled in the art can obtain other drawings based on these drawings without creative effort. 1 shows a schematic view of a typical embodiment according to the present invention, in which a patterned substrate is produced by the filling process and second polymer graft chains are produced in the process; 2a and 2b each show a surface micrograph of an Si substrate with a grown polystyrene graft chain of a typical embodiment according to the present invention, FIG. 2a showing a linearly patterned Si substrate and FIG. 2b a planar patterned substrate; 3 shows a diagram of the infrared spectrum of a filled substrate of a typical embodiment according to the present invention; 4a and 4c each show a liquid phase atomic force microscope image and a structure scanning curve diagram of a planar patterned substrate in ethanol with a grown polystyrene graft chain of a typical embodiment according to the present invention; 4b and 4d each show a liquid phase atomic force microscope image and a structure scanning curve diagram of a planar patterned substrate in DMF with a grown polystyrene graft of a typical embodiment according to the present invention; 5 shows a schematic representation of a typical embodiment according to the present invention, in which the fluorescent groups located at the rear ends are embedded in the planar pattern located on the filling surface by the ATRP reaction in the first step and the nucleophilic substitution reaction in the second step; 6 shows a spectrum diagram of the emission of the fluorescent groups of the labeled macromolecules of a typical embodiment according to the present invention exposed to an excitation light source of 356 nm; 7a and 7b each show a schematic image of the fluorescence intensity of a typical embodiment according to the present invention, in which the second polymer graft chains are in the original state and in the adsorbed state in which the second polymer graft chains are bound to the marked macromolecules.
    Detailed description of the embodiment
    After lengthy research and extensive practice, the inventor proposes with the present invention a technical solution in which the deficiencies known from the prior art are eliminated. The materials that react to the environment can independently select polymer graft chains in order to create adaptable material substrates and to achieve specific reactions to different environmental conditions. The technical solution, the implementation process and the principle are explained in more detail below.
    In one aspect of an embodiment of the present invention, there is provided an environmentally responsive sensing device comprising: a patterned substrate, wherein at least two spaced-apart trenches are formed in the surface of the substrate, the at least two trenches containing initiator first polymers are filled in to form a planar pattern as a detection area on the surface of the substrate; and a plurality of second polymer graft chains, wherein the front ends of the second polymer graft chains are covalently bonded to the planar pattern by reaction with the active functional groups located on the surface of the first polymers, wherein at the same time the rear ends of the second polymer graft chains are covalently bonded to labeled macromolecules, wherein the labeled macromolecules are selected from a group consisting of short block polymers, the short block polymers having fluorescent groups and functional groups which can specifically bind to selected small molecules.
    Further, when the second polymer graft chains are placed in a good solvent, the second polymer graft chains are in the stretched state, and when the second polymer graft chains are placed in a bad solvent, the second polymer graft chains are in the collapsed state, the length of the second polymer graft chains in the stretched state is at least 6 to 10 times the length of this in the collapsed state.
    Further, when the sensing area of the environmentally responsive sensing device is in contact with a good solvent, every plural second polymer graft chains are stretched to form an upright bristle-like structure. When the sensing area of the environmentally responsive sensing device is in contact with a poor solvent, any multiple second polymer graft chains on the surface of the substrate collapse and clear boundaries form between the spaced planar patterns.
    Furthermore, the front ends of the plurality of second polymer graft chains are covalently bonded to active functional groups present on the surface of the first polymers by atom transfer radical polymerization, and the rear ends of the second polymer graft chains are covalently bonded to the labeled macromolecules by a nucleophilic substitution reaction.
    Furthermore, the first polymers comprise one or more than two combinations of polystyrene, polymethyl methacrylate and polyethylene terephthalate, but are not limited thereto.
    Furthermore, the second polymer graft chains are covalently bound to labeled macromolecular proteins, enzymes, nucleic acids, etc. containing amino groups, but are not limited thereto.
    Further, the number average molecular weight of the short block polymers is 300 to 450.
    Furthermore, the short block polymers have a chain segment with 15 to 30 carbon atoms.
    Furthermore, the second polymer graft chains originate from polystyrene, but are not limited thereto.
    Further, the fluorescent groups of the labeled macromolecules are derived from a compound represented by the following formula:
    Furthermore, the planar pattern comprises two or more strip-shaped graphics distributed in parallel.
    Furthermore, the distance between two adjacent strip-shaped graphics is 10 to 2000 nm.
    In a further aspect of the embodiment of the present invention, a method of manufacturing the above-mentioned environment-responsive detection device is further provided, comprising the following steps: (1) providing a patterned substrate, wherein at least two spaced-apart trenches in the Surface of the substrate are formed; (2) Filling the precursor of the active functional group-containing first polymers into the trenches located on the surface of the substrate and then aligning and pressing a plane on the surface of the substrate at the polymerization temperature, the surface energy of the plane being higher than that of the substrate, to form a planar pattern as a detection area on the surface of the substrate; (3) covalently bonding the leading ends of the plurality of second polymer graft chains by atom transfer radical polymerization to the active functional groups on the surface of the first polymers; (4) Covalent attachment of the rear ends of the second polymer graft chains to the labeled macromolecules by a nucleophilic substitution reaction.
    Further comprises step (4): bringing the liquid phase reaction system containing the labeled macromolecules and a good solvent into contact with the detection area and forming the second polymer graft chains in the stretched state and covalently bonding the rear ends of the second polymer graft chains through a nucleophilic substitution reaction on the labeled macromolecules.
    In a further aspect of the embodiment of the present invention, there is further provided a detection method based on the above-mentioned environment-responsive detection device. The detection method includes: bringing the liquid sample to be tested, which contains a poor solvent and selected small molecules, into contact with the detection area of the environment-responsive detection device, and then detecting the change in fluorescence intensity of the detection area by the amount in the detection area to detect selected small molecules in the liquid sample under test.
    The detection process also includes the following:
    Bringing a plurality of standard liquid samples, each containing a poor solvent and selected small molecules in different concentrations, into contact with the detection area of the detection device which is responsive to the environment and detecting the fluorescence intensity of the detection area and establishing the formula F = kλ · c / S showing the relationship between the concentration of selected small molecules and the fluorescence intensity, where F is the fluorescence intensity and the unit is au where kλ is the excitation light source coefficient, where c is the concentration of selected small molecules in the standard liquid sample and has the unit mol / l, where S is the specific surface area of the functional groups of the labeled macromolecules and the recognition area.
    Contacting a liquid sample to be tested containing a poor solvent and selected small molecules in unknown concentration with the detection area of the environmentally responsive detection device and determining the fluorescence intensity of the detection area and calculating the concentration of the selected small molecules the liquid sample to be tested according to the formula.
    Furthermore, the liquid sample to be tested can contain a good solvent. Further, the environmental responsive detection device can accurately respond to the good solvent system and the bad solvent system. Here, the detection object may be an oil-liquid interface solution system or a solid-state system capable of adsorbing various solvents and having an uneven distribution, or a complex biological structure, and so on.
    In the environmentally responsive sensing device of the present invention, the second polymer graft chains are chemically bonded to the first polymers filled in the patterned substrate by atom transfer radical polymerization. Under different environmental conditions, the second polymer graft chains have different swelling effects and change their morphology by reacting to the stimuli of the environment, which leads to a change in the surface structure of the substrate in order to achieve an automatic control function which reacts to the environment.
    The present invention is described in detail below with the aid of a specific exemplary embodiment. It should be pointed out that the exemplary embodiment described below serves to facilitate understanding of the invention and is not intended to limit the protection claims.
    Reference is made to FIG. 1. In one embodiment of the present invention, an active functional group-containing precursor (the ratio of styrene, divinylbenzene and 4-chloromethylstyrene is 7: 1: 2) by filling the polymers in a linearly patterned Si substrate with a length and width of 2 cm x 2 cm and a distance of 500 nm. The substrate is then aligned with a suitable pressure (4 MPa) at the polymerization temperature (110 ° C.) and pressed onto another Si plane with a higher surface energy. Since the Si plane has a higher surface energy than the substrate, polymers with active functional groups can easily be filled into the latter to form a planar patterned substrate in which active functional groups are alternately present and sometimes absent. By changing the concentration of the monomers of the functional groups, the density of the active initiator can be easily adjusted to generate polymer graft chains in certain areas by using ATRP. In the present exemplary embodiment, the polymer graft chains are polystyrene graft chains which contain 40 to 100 carbon atoms and can react to various solvents.
    In the present embodiment, the experimental process of atom transfer radical polymerization is described below: A Si substrate containing the initiating CI groups is placed in a styrene-containing reagent and analytically pure copper (I) chloride, Copper bromide and 2,2'-bipyridine are added (the mass ratio is 7: 1: 28), then after complete mixing they are heated to 120 ° C. in order to produce a Si substrate with CI groups at the rear ends of the polymer graft chains. As shown in FIG. 2, the surface of the Si substrate has changed significantly before and after the reaction and the linearly patterned Si substrate (see FIG. 2a) was converted into a planar patterned substrate (see FIG. 2b) . As shown in the infrared spectrum diagram of Fig. 3, the filled substrate contains CI groups.
    These response characteristics can form a basis for the development of “intelligent” surfaces. Based on the type of interaction with the polymer graft chains, the solvent can be roughly divided into a bad solvent and a good solvent for the polymer graft chains. Due to the interaction between the solvent and the polymer graft chains, the polymer graft chains are always in the collapsed state in the poor solvent. In contrast, in a good solvent they are in the stretched state. The resulting thickness during stretching is controlled by the balance between osmotic pressure and tensile stress. When the quality of the solvent changes from good to bad, the polymer graft chains shrink and gradually change from the stretched state to the collapsed state. Most polymer graft chains can swell 6 to 10 times their state of contraction.
    The liquid phase atomic force microscope (Liquid AFM) and the fluorescence microscope are used to observe the morphological changes in the polymer graft chains in various solvents. Characterization techniques such as scanning electron microscopy and transmission electron microscopy can be used to characterize conventional samples, but the real-time state of the polymer graft chains in the solvent must be examined directly by an in-situ analysis carried out in the solvent. Liquid phase atomic force microscopy (Liquid AFM) and fluorescence microscopy can be performed in a liquid medium / environment to obtain information about the surface morphology of a substrate.
    The inventor used a liquid phase atomic force microscope to examine the in situ state of the polymer graft chains of the present embodiment in dimethylformamide (DMF) and in ethanol, since the use of toluene is not permitted in the liquid phase atomic force microscope. DMF is a good solvent and ethanol is a bad solvent for polymer chains. The changes in conformation of the polymer chains in these two solvents can be very significant (cf. FIGS. 4a to 4d). As shown in FIGS. 4a and 4c, the polymer chains are in the collapsed state in ethanol. Therefore, the height of the polymer graft chains is less than 10 nanometers. As shown in FIGS. 4b and 4d, in the present exemplary embodiment the polymer graft chains in DMF are in the stretched state and their height is approximately 40 nm. At the same time, the width of the polymer graft chains also changes in the present exemplary embodiment. In the dry state, the lateral width of the bristle-like structure of the polymer graft chains is 500 nm, which is greater than its width in the stretched state (approximately 330 nm).
    The characteristics of the environment-reactive polymer graft chains of the present embodiment offer good application possibilities in the field of biological proteins. As shown in FIG. 5, in an application of the present embodiment with reference to the above description, the polymer graft chains are also first formed by ATRP polymerization. The proteins of the marked macromolecules with biological activity (such as Alexa Fluor 350) are then reacted with the polymer graft chains. The reaction process is described below: The Si substrate with the CI groups at the ends of the polymer graft chains is placed in DMF, then Alexa Fluor 350 is added, then the mixture is heated to 60 ° C to attach macromolecular proteins to the ends to bind the polymer graft chains. Since the macromolecular proteins are labeled with fluorescent groups, the state of the polymer graft chains can be easily observed using a fluorescence microscope. The structure of the fluorescent groups of the proteins of the labeled macromolecules is shown below and the emission spectrum measured at a wavelength of 356 nm is shown in FIG.
    By means of the radiation of 356 nm emitted by a light source, the fluorescence emitted by the ends of the polymer graft chains can be observed by means of a fluorescence microscope.
    When using such a detection device, there is no interference from other molecules and the detection device only reacts to certain molecules in the solution. 7a and 7b each show a schematic picture of the fluorescence intensity in which the polymer graft chains are in the original state and in the adsorbed state in which the polymer graft chains are bound to the proteins of the marked macromolecules.
    Therefore, when the detection range of the environment-responsive detection device provided by the present embodiment is placed in a reagent having small molecules, it can be observed that the fluorescence intensity of the surface of the polymer graft chains comprising macromolecular proteins has changed. By measuring this change, the concentration of small molecules in the solution can be determined.
    In particular, specific small molecules (such as amino acids and silver ions) can be specifically adsorbed by the biologically active functional groups located on the surface of the macromolecular proteins. By observing the change in fluorescence intensity on the surface of the polymer graft chains, the number of molecules adsorbed can be determined. As shown in the following formula, once equilibrium is reached, the fluorescence intensity can be used to determine the concentration of specific molecules in the solution:F = kλ * c / S
    Here, F is the fluorescence intensity and has the unit au, where kλ is the excitation light source coefficient, where c is the concentration of selected small molecules in the standard liquid sample and has the unit mol / l, where S is the specific surface area of the functional groups of the labeled macromolecules and of the detection area.
    In a further application, polymer graft chains can be produced using the same ATRP process, then the short block polymers (such as, for example, PdS or polystyrene comprising CdS) by ATRP reaction under the same conditions up to the rear ends of the Modified polymer graft chains.
    In this application, a two-step ATRP process for polymer graft chains is used, with the long polymer chains produced in the first step of the ATRP process, a similar state of swelling can be maintained and the long polymer chains are not greatly changed by the short block polymers, the Thickness of the short block polymers is approximately the same. Therefore, the fluorescence observed by the microscope has a substantially uniform intensity and thus the boundaries of the polymer chains can be clearly observed.
    In summary, with the above technical solution, the polymer graft chain of the detection device according to the invention which reacts to the environment has an elongated conformation in the good solvent and a collapsed conformation in the bad solvent and at the same time enables the screening of certain molecules, which corresponds to an automatic reaction to different environmental conditions , whereby a substrate with sensor materials can be precisely controlled by changing the solvent properties or specific substrates can be designed for measuring the molecular properties of solutions. By adapting the types of material at the rear ends of the polymer graft chains, the detection device can adapt to various environmental conditions, such as e.g. B. different reagent concentrations, different temperatures and different electric and magnetic fields react. In this way a new possibility for developing materials is provided. The invention has the prospect of widespread application in the field of automatic surveillance and also constitutes an important basic information gathering unit for future data development.
    The above description represents only a preferred embodiment of the invention and is not intended to restrict the claims. All equivalent changes and modifications that may be made according to the description and drawings of the invention by one skilled in the art are included within the scope of the present invention.
    权利要求:
    Claims (7)
    [1]
    1. Environment-responsive sensing device characterized in that it comprises:a patterned substrate, wherein at least two spaced-apart trenches are formed in the surface of the substrate, the at least two trenches being filled with initiator-containing first polymers to form a planar pattern as a detection area on the surface of the substrate; andseveral second polymer graft chains and labeled macromolecules, the front ends of the second polymer graft chains being covalently bound by atom transfer radical polymerization to active functional groups present on the surface of the first polymer and to the planar pattern, while at the same time the rear ends of the second polymer graft chains are bound by a nucleophile Substitution reaction are covalently bound to the labeled macromolecules, the labeled macromolecules being selected from a group consisting of block polymers, the block polymers having fluorescent groups and functional groups that can specifically bind to selected molecules, and where a number average molecular weight of the block polymers is 300 to 450, wherein when the second polymer graft chains are further added to a first solvent for the polymer graft chains, the second polymer graft chains are in a stretched state, wherein, we nn the second polymer graft chains are added to a second solvent for the polymer graft chains, the second polymer graft chains are in a collapsed state, wherein the length of the second polymer graft chains in the stretched state is at least 6 to 10 times the length of this in the collapsed state, wherein a type of interaction of the polymer graft chains with the first solvent differs from the type of interaction with the second solvent, furthermore, when the detection area of the environmentally responsive detection device is in contact with the first solvent, all of the plural second polymer graft chains are stretched by to form an upright bristle-like structure wherein, when the sensing area of the environmentally responsive sensing device is in contact with the second solvent, all of the plurality of second polymer grafts on the surface of the substrate are collapsed and between the there are clear boundaries between spaced planar patterns.
    [2]
    2. Detection device which reacts to the environment according to claim 1, characterized in thatthe first polymers comprise one or more than two combinations of polystyrene, polymethyl methacrylate, and polyethylene terephthalate;and / or the second polymer graft chains are covalently bonded to labeled macromolecules containing amino groups; wherein the labeled macromolecules comprise one or more than two combinations of proteins, enzymes and nucleic acids;the block polymers preferably having a chain segment with 15 to 30 carbon atoms.
    [3]
    3. Environmentally responsive detection device according to claim 1, characterized in that the second polymer graft chains are derived from polystyrene; and / or the fluorescent groups of the labeled macromolecules are derived from a compound represented by the following formula:
    [4]
    4. Environment-responsive detection device according to claim 1, characterized in that the planar pattern comprises two or more strip-shaped graphics distributed in parallel; wherein the distance between two adjacent strip-shaped graphics is preferably 10 to 2000 nm.
    [5]
    5. The method for producing the sensing device which is responsive to the environment according to any one of claims 1 to 4, characterized in that it comprises the following steps:1) providing a patterned substrate, wherein at least two spaced-apart trenches are formed in the surface of the substrate;2) Filling the precursor of the active functional group-containing first polymers into the trenches located in the surface of the substrate, and then aligning and pressing a plane on the surface of the substrate at the polymerization temperature, the surface energy of the plane being higher than that of the substrate forming a planar pattern as a detection area on the surface of the substrate;3) Covalent bonding of the front ends of the plurality of second polymer graft chains by atom transfer radical polymerization to the active functional groups located on the surface of the first polymers;4) Covalent binding of the rear ends of the second polymer graft chains to the labeled macromolecules by a nucleophilic substitution reaction,wherein step 4) comprises: bringing a liquid phase reaction system containing the labeled macromolecules and a solvent into contact with the detection area and forming the second polymer graft chains in the stretched state and covalently bonding the rear ends of the second polymer graft chains by a nucleophilic substitution reaction to the labeled macromolecules.
    [6]
    6. Detection method, which is based on the detection device according to one of claims 1 to 4, which is responsive to the environment, characterized in that the detection method comprises the following: bringing into contact a liquid sample to be tested which contains a solvent and selected molecules, with the detection area of the environment-responsive detection device and then detecting the change in fluorescence intensity of the detection area to detect the selected molecules present in the liquid sample to be tested.
    [7]
    7. The detection method according to claim 6, characterized in that it comprises:Bringing a plurality of standard liquid samples, each containing a first solvent and selected molecules in different concentrations, into contact with the detection area of the detection device which reacts to the environment and detecting the fluorescence intensity of the detection area and establishing the formula F = kλ · c / S, which shows the relationship between the concentration of the selected molecules and the fluorescence intensity, where F is the fluorescence intensity and the unit is au where kλ is the excitation light source coefficient, where c is the concentration of the selected molecules in the standard liquid sample and has the unit mol / l, where S is the specific surface area of the functional groups of the labeled macromolecules and the recognition region;Contacting a liquid sample to be tested containing a first solvent and selected molecules in unknown concentration with the detection area of the environmentally responsive detection device and determining the fluorescence intensity of the detection area and calculating the concentration of the selected small molecules of the liquid to be tested Sample according to the formula;wherein preferably the liquid sample to be tested further comprises a second solvent.
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    法律状态:
    优先权:
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