WO2023104012A1

WO2023104012A1 - Resonant chip and manufacturing method therefor

Info

Publication number: WO2023104012A1
Application number: PCT/CN2022/136825
Authority: WO
Inventors: 李毅; 杨亚涛
Original assignee: 南方科技大学
Priority date: 2021-12-06
Filing date: 2022-12-06
Publication date: 2023-06-15
Also published as: CN114156625A; CN115939718A

Abstract

The present application relates to the field of semiconductors, and relates to a resonant chip and a manufacturing method therefor. The resonant chip comprises: a dielectric layer. A resonant channel is provided on the dielectric layer; the dielectric layer comprises a resonator and a transitional dielectric layer; a resonator is formed on the surface of the transitional dielectric layer; the resonant channel sequentially passes through the resonator and the transitional dielectric layer; the resonator is used for forming a resonance response with excitation light, and confining electric field energy in the resonant channel; the transitional dielectric layer is used for blocking the passage of biomolecules, so that a biological reaction only occurs in the resonant channel. Compared with a metal cladding layer, the use of the dielectric layer in the present application avoids the problems of energy loss of electromagnetic waves and light quenching. The resonant channel is placed into the resonator to bind an electromagnetic field in the resonant channel, so that a single-molecule fluorescence process in the resonant channel is enhanced. The biological reaction only occurs in the resonant channel, so that the probability of capturing reactants is controlled and improved.

Description

Resonator chip and manufacturing method thereof

Cross References to Related Applications

This application requires the application number 202211538318.6 submitted to the China Patent Office on December 1, 2022, titled "A Resonant Chip and Its Manufacturing Method" and the application number 202111478357.7 submitted to the China Patent Office on December 6, 2021, title It is the priority of the Chinese patent application "a resonator chip and its manufacturing method", the entire content of which is incorporated in this application by reference.

technical field

The present application relates to the field of semiconductors, in particular, to a resonant chip and a manufacturing method thereof.

Background technique

The combination of solid-state electronics technology and biological research applications has made many important advances, including molecular array technology, microfluidic chip technology, chemically sensitive field-effect transistors, and valuable sensing technologies such as zero-mode waveguides.

Molecular array technology, namely DNA array (US patent 6261776), microfluidic chip technology (US patent 5976336), chemically sensitive field effect transistor, zero mode waveguide (Chinese patent CN101467082B), and other valuable sensing technologies.

Zero-mode waveguide (ZMW) arrays have extended semiconductor fabrication technology further into research and diagnostics, and have been used in a range of biochemical analyses, especially in the field of genetic analysis. Typical ZMWs consist of openings, wells, or nanoscale cores in an opaque cladding on a transparent substrate. The narrow dimensions of the core consistently prevent electromagnetic radiation with frequencies above a certain cutoff frequency from propagating through it, so that by irradiating very small volumes very small quantities of reactants, including unimolecular reactions, can be accessed.

By monitoring reactions at the single-molecule level, the ability to precisely identify and/or monitor a given reaction is fundamental in the field of single-molecule DNA sequencing technology—molecular synthesis of DNA strands by a single DNA polymerase in a template-dependent manner to monitor molecules .

However, when the existing array chips perform biochemical fluorescence analysis, there are often problems of low accuracy of fluorescence sequencing and low signal-to-noise ratio of fluorescence signals.

Currently, metallic ZMW arrays are usually used with relatively low excitation field strengths in the core, however, this requires high input power to meet the signal-to-noise ratio for single-molecule detection. High input power will increase the error rate of DNA polymerase, and at the same time, it is easier to produce photochemical processes such as photobleaching of bioluminescent molecules, introducing unnecessary optical interference and errors.

In the process of preventing the propagation of electromagnetic wave radiation, ZMW will produce a large amount of electromagnetic wave energy loss. Under the premise of high input power, these energy losses will be transmitted to the system in the form of heat, thereby affecting the biochemical stability of the substrate, affecting the activity and error rate of DNA polymerase.

Since ZMWs are mainly openings or wells on the metal film with large light loss, there is a problem of photoquenching of the fluorescence signal during the radiation process, that is, the relative distance between the enzyme reaction site of the radiated fluorescence and the metal core will seriously affect the single The intensity of the molecular fluorescent radiation.

Most of the above problems are determined by the material system selected for the substrate. The biochemical stability of the substrate can be coated by coating or monolayer surface modification. However, conventional structural changes have not fundamentally solved the problems of high input power, temperature rise, and photoquenching, that is, there is no effective solution to the interference and error increase introduced by the enzymatic process.

Contents of the invention

The purpose of the embodiments of the present application is to provide a resonant chip and a manufacturing method thereof.

In a first aspect, the present application provides a resonant chip, including:

a dielectric layer, a resonant channel is arranged on the dielectric layer, and the resonant channel penetrates the dielectric layer;

The dielectric layer includes a resonator and a transition dielectric layer;

The resonator is formed on the surface of the transition dielectric layer;

The resonance channel runs through the resonator and the transition dielectric layer in turn; the resonator is used to form a resonance response with the excitation light, and the electric field energy is confined in the resonance channel; the transition dielectric layer is used to isolate the biomolecules from passing through, so that the biological reaction is only in the resonance channel internal reaction.

First of all, the resonant chip of the present application is provided with a dielectric layer with low optical loss and high refractive index, which avoids the energy loss of electromagnetic waves and the problems of optical quenching compared with the metal cladding layer of the existing ZMW.

Secondly, the present application proposes and designs a resonator with a resonant channel. The design principle is different from that of the ZMW, which prevents electromagnetic radiation with a frequency higher than a specific cut-off frequency from propagating through the core. Instead, the electromagnetic mode interaction of the resonator will The energy of the light field is bound in the resonance channel to realize the enhancement of the single-molecule fluorescence process in the resonance channel.

Most importantly, the design of the chip's two-layer dielectric layer, in which the resonator, is used to identify and/or monitor a given reaction; to form a resonance response with the excitation light, and to confine the electric field energy in the resonance channel; the transition dielectric The layer is used to isolate the passage of biomolecules, so that the biological reaction only reacts in the resonance channel, thereby controlling and improving the capture probability of the reactant.

In other embodiments of the present application, the above-mentioned resonator is island-shaped.

In other embodiments of the present application, the edges of the above-mentioned resonators form a regular shape or a topological shape.

In other embodiments of the present application, the above-mentioned resonant channels are in the shape of slits, regular holes or irregular holes.

In other embodiments of the present application, the maximum value of the narrowest width of the above-mentioned resonant channel is 20 nm.

In other embodiments of the present application, the total volume of the above-mentioned resonant channel is controlled at the single-molecule detection level.

In other embodiments of the present application, the above-mentioned dielectric layer is made of non-metallic materials.

In other embodiments of the present application, the materials of the above-mentioned resonator and the transition dielectric layer may be the same or different.

In other embodiments of the present application, the above-mentioned resonant chip further includes:

Substrate;

Conductive layer formed on the surface of the substrate.

In other embodiments of the present application, the above-mentioned conductive layer is made of electrode material for external voltage.

In other embodiments of the present application, the above resonant chip further includes: an adhesion layer formed on the surface of the conductive layer; and a dielectric layer formed on the surface of the adhesion layer.

In other embodiments of the present application, the above-mentioned adhesion layer is made of porous material.

In other embodiments of the present application, the above-mentioned resonant chip includes water seepage holes; the water seepage holes pass through the substrate and the conductive layer, and are in contact with the adhesion layer.

The application provides a resonant chip, and the resonant chip includes:

Substrate;

a conductive layer on one side of the substrate;

an adhesion layer on one side of the conductive layer;

A dielectric layer located on one side of the adhesion layer, wherein a resonant slit is arranged on the dielectric layer, and the resonant slit penetrates through the dielectric layer.

In other embodiments of the present application, the above-mentioned dielectric layer includes a resonator and a transition dielectric layer;

The transition dielectric layer is located on one side of the adhesion layer, and the resonator is located on one side of the transition dielectric layer;

The resonant slits sequentially penetrate the resonator and the transition dielectric layer.

In other embodiments of the present application, the above-mentioned resonator is in a regular or irregular shape such as a circle or a square, and the shape of the resonant slit is a strip-shaped rectangle, and the resonant slit is arranged at the center of the resonator.

In other embodiments of the present application, the above-mentioned materials for making the resonator and the transition dielectric layer include non-metallic materials such as gallium phosphide and gallium nitride.

In other embodiments of the present application, the above-mentioned dielectric layer further includes a dielectric waveguide, and the dielectric waveguide is located on the transition dielectric layer.

In other embodiments of the present application, the light beam emitted by the above-mentioned dielectric waveguide enters the resonator horizontally.

In other embodiments of the present application, the light beam emitted by the above-mentioned dielectric waveguide is parallel to the resonator.

In other embodiments of the present application, the above-mentioned conductive layer and the substrate are further provided with water seepage holes, and the positions of the water seepage holes correspond to the resonant slits.

In a second aspect, the present application provides a method for manufacturing a resonant chip, the method comprising:

setting a resonant channel on the dielectric layer, so that the resonant channel penetrates the dielectric layer;

The dielectric layer includes a resonator and a transition dielectric layer, so that the resonator is formed on the surface of the transition dielectric layer; the resonant channel passes through the resonator and the transition dielectric layer in turn; the resonator is used to form a resonance response with the excitation light, and make the electric field energy Confined in the resonant channel; the transition dielectric layer is used to isolate biomolecules from passing through, so that biological reactions only react in the resonant channel.

In other embodiments of the present application, the present application provides a method for manufacturing a resonant chip, the method comprising:

providing a substrate;

generating a conductive layer along one side of the substrate;

Create an adhesion layer along one side of the conductive layer;

A dielectric layer is fabricated along one side of the adhesion layer, wherein a resonant slit is arranged on the dielectric layer, and the resonant slit penetrates through the dielectric layer.

Description of drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present application, the accompanying drawings that are required in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present application, and thus It should be regarded as a limitation on the scope, and those skilled in the art can also obtain other related drawings based on these drawings without creative work.

FIG. 1 is one of the cross-sectional schematic diagrams of a resonant chip provided in an embodiment of the present application;

Fig. 2 is the second schematic cross-sectional view of the resonant chip provided by the embodiment of the present application;

FIG. 3 is one of the structural schematic diagrams of the resonant chip provided by the embodiment of the present application;

Fig. 4 is the third schematic cross-sectional view of the resonant chip provided by the embodiment of the present application;

FIG. 5 is a perspective view of a resonant chip provided in an embodiment of the present application;

FIG. 6 is the second structural schematic diagram of the resonant chip provided by the embodiment of the present application;

FIG. 7 is the third structural schematic diagram of the resonant chip provided by the embodiment of the present application;

FIG. 8 is a flow chart of a manufacturing method of a resonant chip provided in an embodiment of the present application;

FIG. 9 is the second flow chart of the manufacturing method of the resonant chip provided by the embodiment of the present application;

Fig. 10 is the gallium phosphide resonator chip provided by the embodiment of the present application; wherein, in Fig. 10, (a) the top view of the scanning electron microscope, (b) the near-field focusing distribution of electromagnetic waves under the condition of 459nm wavelength light excitation, (c) 584nm wavelength The near-field focusing distribution of electromagnetic waves under the condition of light excitation;

Figure 11 is the preliminary calculation and processing diagram of the gallium phosphide resonator chip provided by the embodiment of the present application, (a) scattering spectrum, (b) absorption spectrum, (c) electromagnetic field enhancement performance of the resonator under different slit lengths; d) Processed gallium phosphide resonator chips with different slit sizes;

Figure 12 is the preliminary verification of the chip provided by the embodiment of the present application and the comparative example; (a) the absorption and scattering spectrum comparison of the gallium phosphide resonator and the ZMW; (b) the electromagnetic field in the slit of the gallium phosphide resonator and in the hole of the ZMW enhanced performance;

Figure 13 is the calculation verification of the dielectric resonant chip provided by the embodiment of the present application and the comparative example; (a) the enhanced fluorescence quantum yield comparison of ZMW and gallium phosphide resonant chip; (b) the fluorescence of ZMW and gallium phosphide resonant chip Radiation Enhanced Contrast;

Figure 14 is the computational verification of the chip-enhanced fluorescence provided by the examples and comparative examples of the present application; (a) the distribution in the hole of the ZMW fluorescence radiation; (b) the distribution in the slit of the fluorescence radiation of the gallium phosphide resonator; (c) In-slit distribution of fluorescence radiation of GaN resonators;

Figure 15 is the preliminary verification of the enhanced excitation of the dielectric waveguide resonator chip provided by the embodiment of the present application; (a) schematic diagrams of four excitation modes; (b) electric field enhancement spectrum under corresponding excitation modes; (c) corresponding excitation modes Electric field distribution map;

Figure 16 is the preliminary calculation and verification of the enhanced quantum yield of the dielectric waveguide resonant chip provided by the embodiment of the present application; (a) schematic diagram of different units on the waveguide resonant chip, and (b) when the initial quantum yield is 0.3 and (c) The fluorescence quantum yield enhancement spectrum when the initial quantum yield is 0.003. (d) Schematic diagram of on-chip collection and radiation directionality.

Icons: 101-dielectric layer; 102-adhesion layer; 103-conductive layer; 104-substrate; 1011-resonator; 1012-transition dielectric layer; 1013-dielectric waveguide.

Detailed ways

In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present application, not all the embodiment.

Accordingly, the following detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but merely represents selected embodiments of the present application. Based on the embodiments in this application, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of this application.

As recorded in the background technology, many important advances have been made in the combination of solid-state electronics technology and biological research applications, including molecular array technology, microfluidic chip technology, chemically sensitive field-effect transistors, and zero-mode waveguides. Sensing Technology.

The problems existing in the prior art are all the results obtained by the inventor after practice and careful research. Therefore, the discovery process of the above problems and the solutions to the above problems proposed by the embodiments of the present application below should all be It is the contribution made by the inventor during the invention process.

In view of this, in order to solve the above problems, this application provides a resonant chip. By setting a resonant channel, the fluorescence enhancement effect of biomolecules in the channel is greatly improved, thereby effectively improving the accuracy of fluorescent sequencing and the signal of fluorescent signals. noise ratio.

Referring to FIGS. 1-7 , some embodiments of the present application provide a resonant chip, including: a dielectric layer 101 . A resonant channel is provided on the dielectric layer 101 , and the resonant channel penetrates the dielectric layer 101 .

The above-mentioned resonant channel is the resonant slit in FIG. 1 . When the resonator forms a resonance response with the excitation light, the electric field energy can be confined in the resonance channel. The resonant slit in FIG. 1 is also called a resonant channel, that is, the electric field energy can also be confined in the resonant slit in FIG. 1 .

Further, the above dielectric layer 101 includes a resonator 1011 and a transition dielectric layer 1012 .

Further, the resonator 1011 is formed on the surface of the transition dielectric layer 1012 .

Further, the resonant channel runs through the resonator 1011 and the transition dielectric layer 1012 in sequence; the resonator 1011 is used to form a resonance response with the excitation light, and makes the electric field energy confined in the resonant channel; the transition dielectric layer 1012 is used to isolate biomolecules from passing through , so that the biological response is only in the resonant channel.

The resonant chip of this application utilizes a high-refractive index dielectric resonator 1011 and a resonant channel structure, which has rich electromagnetic mode characteristics in the visible light band, and the interaction between the electrode and the ring pole mode at a specific wavelength makes the near-field energy Confined in the resonant channel of the resonator 1011, the far-field scattering appears as a non-polar sub-mode. At this time, the electric field is all concentrated in the resonant channel, which improves the local electric field intensity. Quantum yield, and achieve directional radiation, which is beneficial to the collection of fluorescent signals.

Further, in some embodiments of the present application, the resonator 1011 is island-shaped.

The above-mentioned island-shaped resonator 1011 corresponds to the disc-shaped resonator 1011 in FIG. 3 . In other words, the disk-shaped resonator 1011 in FIG. 3 may also be called an island-shaped resonator 1011 . That is, the resonators protruding from the transition dielectric layer 1012 can be used to form a resonance response with the excitation light.

Further, in some embodiments of the present application, the edges of the resonator 1011 form a regular shape or a topological shape.

In other words, the overall shape of the resonator 1011 of the present application is not limited. For example, in some specific implementation manners, the above-mentioned island-shaped resonator 1011 may be disc-shaped; the bottom edge may be disc-shaped, but there are protrusions on the surface of the disc-shaped bottom surface, forming an island shape . It is also possible that the bottom edge is quadrilateral or other irregular shape, such as a topological shape, but there are protrusions on the upper surface of the quadrilateral or other irregular bottom surface, forming an island shape.

Further, in some embodiments of the present application, the maximum width of the narrowest part of the resonant channel is 20 nm. Within this range, the local electric field intensity inside the resonant channel can be greatly increased, and at the same time, the quantum yield of the auxiliary enhanced fluorescent molecules in the process of fluorescent radiation can be improved.

Further optionally, in some embodiments of the present application, the width of the resonant channel is 0.1 nm˜19.8 nm. Further optionally, in some embodiments of the present application, the width of the resonant channel is 1 nm˜19 nm. Exemplarily, the width of the resonance channel is 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 11 nm, 12 nm, 13 nm, 14 nm, 15 nm, 16 nm, 17 nm, 18 nm or 19 nm.

Further, in some embodiments of the present application, the total volume of the resonance channel is controlled at the level of single-molecule detection. By controlling the total volume of the resonant channel at the level of single-molecule detection, it is possible to ensure that single molecules enter the resonant channel and improve the quantum yield of auxiliary enhanced fluorescent molecules in the process of fluorescent radiation.

Exemplarily, the detection level of the above-mentioned single molecule is to use the slit channel to realize the detection volume of the biological solution to the order of 10-21L.

Further, in some embodiments of the present application, the dielectric layer 101 is made of a dielectric material.

Further optionally, in some embodiments of the present application, the above-mentioned dielectric layer 101 is made of dielectric materials such as gallium phosphide and gallium nitride.

In other optional implementation manners of the present application, the above-mentioned dielectric layer 101 may also be made of other common dielectric materials in the field.

Further, in some embodiments of the present application, the materials of the resonator 1011 and the transition dielectric layer 1012 may be the same or different.

Exemplarily, in some embodiments of the present application, the materials of the resonator 1011 and the transition dielectric layer 1012 are gallium phosphide; that is, the materials of the resonator 1011 and the transition dielectric layer 1012 are both made of gallium phosphide.

In other optional implementation manners of the present application, the materials of the resonator 1011 and the transition dielectric layer 1012 are different. Exemplarily, materials of the resonator 1011 and the transition dielectric layer 1012 are gallium phosphide and gallium nitride, respectively. That is, the resonator 1011 is made of gallium phosphide; the transition dielectric layer 1012 is made of gallium nitride.

Further, in some embodiments of the present application, the resonant chip includes: a dielectric layer 101, a substrate 104, a conductive layer 103 formed on the surface of the substrate, and an adhesion layer 102 formed on the surface of the conductive layer 103; the dielectric layer 101 is formed on the surface of the adhesion layer 102.

The dielectric layer 101 here can be the dielectric layer 101 provided in any of the foregoing embodiments.

This chip adopts the design of at least two cladding layers, one of which is a dielectric layer 101, a continuous array of dielectric resonators, used to identify and/or monitor a given reaction; another layer of adhesion layer 102, a porous transport structure , DC electrophoresis or dielectrophoresis can be used to control and improve the capture probability of reactants.

Further, in some embodiments of the present application, the conductive layer 103 is made of electrode material for external voltage.

Further optionally, in some embodiments of the present application, the conductive layer 103 may be made of a dielectric film (silicon nitride) or a conductive film (indium tin oxide, ITO).

In other optional implementation manners of the present application, the above-mentioned conductive layer 103 may also be made of other common conductive materials in the field.

Further, in some embodiments of the present application, the adhesion layer 102 is made of a porous material, which is used to attach biomolecules to the surface of the layer, and is also used to ensure the water permeability of biomolecular tests in a liquid environment, enhancing Liquid in and out, improve the fluidity of the liquid.

Further optionally, in some embodiments of the present application, the adhesion layer 102 is made of a porous aluminum oxide or titanium oxide film.

In some embodiments of the present application, the material of the above-mentioned adhesion layer 102 can be made of other common hole materials in the field.

Further, in some embodiments of the present application, the resonant chip includes a water seepage hole; the water seepage hole penetrates the substrate 104 and the conductive layer 103 , and is in contact with the adhesion layer 102 .

The above water seepage hole corresponds to the water seepage hole in FIG. 4 . By providing water seepage holes, the liquid in the resonant slit can pass through the adhesion layer 102 and soak into the water seepage holes, thereby improving the fluidity of the liquid.

Some embodiments of the present application provide a method for manufacturing a resonant chip, the method including:

Further, in some specific embodiments of the present application, the above-mentioned resonant chip is prepared according to the following steps:

High-quality III-V material thin films and nitride material thin films can be grown on lattice-matched substrates by means of epitaxial deposition. Its typical processing technology is shown in Figure 9 (the direction of the arrow is the direction of the preparation step). First, the adhesion layer 102 (for example, a porous aluminum oxide or titanium oxide film) can be covered on the surface of the sandwich substrate by atomic layer deposition. (For example, AlGap sandwiched between Gap substrates in the figure); Secondly, the conductive layer 103 (dielectric film (for example, silicon nitride) or conductive film (for example, indium tin oxide, ITO)) can be deposited by physical deposition or chemical deposition The method is formed on another substrate 104 (substrate, such as silicon dioxide); again, the two substrates are assembled together by bonding, and the chemical etching method selectively removes the sacrificial layer (for example, the sandwich in the figure AlGaP or AlGaN). Then a dielectric layer 101 is deposited on the surface of the adhesion layer 102 to form a resonator 1011 . III-V or nitride single crystal substrates can be reused. After bonding the III-V or nitride thin films, the required patterns and via holes are formed by two steps of exposure and etching. Finally, you can choose to open the support layer of the substrate by long-time etching to obtain the inlet of the reaction solution, as shown in Figure 9.

In other optional embodiments of the present application, the porous dielectric resonator can also choose other high refractive index films (such as titanium dioxide TiO2, chalcogenide glass Sb2S3, etc.), these materials have lower quality requirements, and can usually be obtained by evaporation and Sputtering and other processing methods, so it can be directly formed on the substrate containing porous alumina or titania. Subsequent photolithography (or nanoimprinting) and etching processes are the same as the aforementioned III-V and nitride systems.

Further optionally, in other optional embodiments of the present application, the processed device can be assembled by plasma oxygen cleaning to form a fluid chamber for the solution to interact with the porous dielectric resonator. Devices with fluids can be placed on an inverted microscope or CMOS camera for imaging of single-molecule fluorescence signals. At the same time, the electrodes and fluid channels carried by the chip can be connected with electrical controls and detectors to control the process of charged biological single molecules entering and exiting the slit.

Compared with the conventional ZMW, this application proposes and uses a dielectric material with no light loss and high refractive index as the cladding layer of the substrate, completely avoiding the use of the metal cladding layer, thereby avoiding the energy loss of electromagnetic waves and optical quenching The problem.

This application proposes and designs a resonator with a resonant channel. The design principle is different from that of ZMW, which prevents electromagnetic radiation with a frequency higher than a specific cut-off frequency from propagating through the core. Instead, the resonator traps the electromagnetic field in the slit to achieve For the enhancement of single-molecule fluorescence processes within the slit.

Some embodiments of the present application provide a resonant chip.

Please refer to FIG. 1, as an optional implementation mode, the resonant chip includes a substrate 104, a conductive layer 103, an adhesive layer 102, and a dielectric layer 101; wherein, the conductive layer 103 is located on one side of the substrate 104, and the attached The layer 102 is located on one side of the conductive layer 103 , and the dielectric layer 101 is located on one side of the adhesion layer 102 , that is, the substrate 104 , the conductive layer 103 , the adhesion layer 102 and the dielectric layer 101 are connected layer by layer. Wherein, a resonant slit is arranged on the dielectric layer 101 , and the resonant slit penetrates through the dielectric layer 101 .

In this embodiment, the substrate 104 is used as the supporting layer of the chip to fix the conductive layer 103 and ensure the stability of the structure of each level unit above it; an appropriate voltage can be applied to the conductive layer 103 through an external device to accelerate the sample to be tested. Biologically specific reaction at the resonance slit; the adhesion layer 102 is also referred to as the bioadhesion water-permeable layer, which is used to attach biomolecules to the surface of the layer, and is also used to ensure the water permeability of biomolecular tests in a liquid environment It can strengthen the liquid in and out, and improve the fluidity of the liquid; the dielectric layer 101 is made of all-dielectric nanomaterials, and the dielectric layer 101 is provided with a penetrating resonant slit, thereby forming a resonant unit. Next, the dielectric layer 101 can confine the light field energy in the dielectric layer 101, because the existence of the resonant slit breaks the original polarization mode, so that the light field energy is all concentrated inside the resonant slit, realizing the local field The enhancement is conducive to improving the luminous efficiency of fluorescent molecules inside the resonance slit, that is, fluorescence enhancement, thereby effectively improving the accuracy of fluorescence sequencing and the signal-to-noise ratio of fluorescence signals.

The resonant chip provided in this implementation will be described in detail below by taking the fluorescence test of biomolecules as an example.

When using the resonant chip to perform fluorescence tests on biomolecules, it is necessary to immerse the resonant chip in a liquid environment. There are corresponding biomolecules in the liquid environment, and the biomolecules will slowly move into the resonance slit and adhere to the adhesion layer 102 At this time, by irradiating a beam of corresponding wavelength, due to the existence of the resonance slit, the energy of the light field will be concentrated inside the resonance slit, thereby achieving fluorescence enhancement and improving the accuracy of fluorescence sequencing and the signal-to-noise ratio of fluorescence signals the goal of. At the same time, since the adhesion layer 102 itself has a certain degree of permeability, it can also improve the fluidity of the liquid environment. When it is necessary to observe the specific reaction of biomolecules, it can be achieved by applying an appropriate voltage to the conductive layer 103 .

Referring to FIG. 2 , as another optional implementation manner, the dielectric layer 101 includes a resonator 1011 and a transition dielectric layer 1012 .

The transition dielectric layer 1012 is located on one side of the adhesion layer 102, and the resonator 1011 is located on one side of the transition dielectric layer 1012, that is, the adhesion layer 102, the transition dielectric layer 1012 and the resonator 1011 are arranged layer by layer.

The resonant slots pass through the resonator 1011 and the transition dielectric layer 1012 in turn.

In this embodiment, the resonator 1011 and the transition dielectric layer 1012 are made of the same kind of dielectric material, which is essentially integrally formed. The function of the transition dielectric layer 1012 is mainly to isolate the adhesion layer 102 from the liquid environment, so that biomolecules can attach to the adhesion layer 102 through the resonant slit, and the function of the resonator 1011 is mainly to absorb light field energy.

It should be noted that when the dielectric layer 101 has a planar structure, the resonator 1011 and the transition dielectric layer 1012 are located on the same plane, and the two are integrated to form the dielectric layer 101 . However, when the dielectric layer has a convex structure, when the dielectric layer 101 is divided into a resonator 1011 and a transition dielectric layer 1012 , it is essentially another structure of the dielectric layer 101 .

Referring to FIG. 3 , in another possible implementation manner, the resonator 1011 is circular, the shape of the resonant slit is a strip-shaped rectangle, and the resonant slit is arranged at the center of the resonator 1011 .

It should be noted that, specifically, the resonator 1011 is in the shape of a disk, and the resonant slit is a narrow long rectangle. The center point of the resonant slit is perpendicular to the center of the resonator 1011. 1011 and transition dielectric layer 1012. The long resonant slit can better improve the effect of fluorescence enhancement.

In another possible embodiment, the material for making the resonator 1011 and the transition dielectric layer 1012 includes gallium phosphide.

It should be noted that the materials of the above-mentioned resonator 1011 and transition dielectric layer 1012 are only one of the implementation modes, including but not limited to gallium phosphide, or other dielectric materials with no optical loss and high refractive index. Therefore, it is beneficial to avoid the loss of light field energy and the problems of light quenching.

In another optional implementation manner, the refractive index of the dielectric layer 101 is higher than the refractive index of the adhesion layer 102, that is to say, the material of the dielectric layer 101 should have a refractive index higher than that of the adhesion layer 102 material. dielectric material.

Since the biological sample is in a liquid environment, in order to improve the fluidity of the liquid environment, in another optional implementation, please refer to FIG. 4 and FIG. The seepage hole, the position of the seepage hole corresponds to the resonance slit.

The position of the water seepage hole corresponds to the resonance slit, which means that the water seepage hole should be set directly below the resonance slit, and the center point of the water seepage hole and the center point of the resonance slit are located on the same vertical axis.

By providing water seepage holes, the liquid in the resonant slit can pass through the adhesion layer 102 and soak into the water seepage holes, thereby improving the fluidity of the liquid.

Generally speaking, when the spatial light is vertically incident on the surface of the resonant chip, the polarization mode will affect the spatial distribution of its electric field, and the energy of the light field at a specific wavelength is bound inside the slit to achieve the enhancement of molecular fluorescence in the slit. This vertical excitation system is difficult to achieve optical excitation on the chip, which is not conducive to the construction of an integrated optical system.

In view of this, please refer to FIG. 6 , in another possible implementation manner, the dielectric layer 101 further includes a dielectric waveguide 1013 , and the dielectric waveguide 1013 is located on the transition dielectric layer 1012 .

It should be noted that, in this embodiment, the dielectric waveguide is located on the transition medium layer, and the dielectric waveguide should be located on the same plane as the resonator 1011, and the thickness of the two should be the same. By setting the dielectric waveguide, parallel incident resonance of excitation light can be realized device 1011, so as to realize higher-energy light field localization in the resonance slit, greatly improve the molecular fluorescence enhancement effect in the resonance slit, and realize the construction of a portable integrated optical system.

In an optional embodiment, please continue to refer to FIG. 6 , the light beam emitted by the dielectric waveguide 1013 is horizontally incident on the resonator 1011 (as shown by the arrow in the figure, which is the direction in which the light beam is emitted), so as to realize the horizontal incident of the excitation light.

It should be noted that the horizontal incidence in this embodiment means that when the dielectric waveguide 1013 and the resonator 1011 are located on the same plane, the light beam emitted by the dielectric waveguide 1013 passes through the side of the resonator 1011 (that is, perpendicular to the transition dielectric layer 1012) is incident on the resonator 1011.

It should be noted that, in practical applications, the position of the dielectric waveguide 1013 can be adjusted as required, so as to adjust the direction of the light beam emitted by it.

In another optional implementation manner, please refer to FIG. 7 , the light beam emitted by the dielectric waveguide 1013 is parallel to the resonator 1011 (as shown by the arrow in the figure, which is the emission direction of the light beam).

In this embodiment, although the dielectric waveguide 1013 is still located on the same plane as the resonator 1011, the light velocity emitted by the dielectric waveguide 1013 does not enter the resonator 1011 through the side of the resonator 1011, but is parallel to the entire resonator 1011 , the transmission form of total internal reflection of the light beam in the dielectric waveguide 1013 leads to the generation of evanescent waves at the surface of the waveguide, when the resonator 1011 is close to the side of the dielectric waveguide 1013, the energy of the evanescent wave at the surface of the waveguide can be coupled into the resonator 1011 Because the existence of the resonant slit makes the electric field trapped in the resonant slit, the molecular fluorescence in the resonant slit can be enhanced, and the coupling efficiency can be improved by setting the distance between the side of the dielectric waveguide 1013 and the resonator 1011.

It should be noted that, in this embodiment, the length of the dielectric waveguide 1013 can be set arbitrarily. In this embodiment, the dielectric waveguide 1013 is preferably extended to the edge of the chip.

Please refer to FIG. 8, the embodiment of the present application also provides a method for manufacturing a resonant chip, the method includes the following steps:

Step 201: providing a substrate 104;

Step 202: generating a conductive layer 103 along one side of the substrate 104;

Step 203: generating the adhesion layer 102 along one side of the conductive layer 103;

Step 204 : Fabricate a dielectric layer 101 along one side of the adhesion layer 102 , wherein a resonant slit is arranged on the dielectric layer 101 , and the resonant slit penetrates through the dielectric layer 101 .

In another optional implementation manner, after the above step 204, the method further includes:

Step 205: Divide the dielectric layer 101 into a resonator 1011 and a transition dielectric layer 1012, the transition dielectric layer 1012 is located on one side of the adhesion layer 102, the resonator 1011 is located on one side of the transition dielectric layer 1012, and the resonance slits are sequentially through the resonator 1011 and the transition dielectric layer 1012 .

Optionally, in the above step 205, the resonator 1011 is circular, the shape of the resonant slit is a strip-like rectangle, and the resonant slit is arranged at the center of the resonator 1011.

Below in conjunction with embodiment, feature and performance of the present application are described in further detail:

Example 1

A gallium phosphide resonator chip is provided, and the structure is shown in FIG. 1 .

An indium tin oxide conductive layer 103 is formed on the surface of the substrate 104 of silicon dioxide, a porous aluminum oxide adhesion layer 102 is formed on the surface of the conductive layer 103, a transition dielectric layer 1012 is formed on the surface of the adhesion layer 102, and a transition dielectric layer 1012 is formed on the surface A disk-shaped resonator 1011 is formed. The resonant channel is slit-shaped. Both the transition dielectric layer 1012 and the resonator 1011 are formed of gallium phosphide.

Comparative example 1

Provide common chips in the prior art: ZMW.

The performance of the chip of Example 1 and Comparative Example 1 is tested as follows, and the results (Fig. 10-Fig. 16) are calculated by using the finite time domain difference method for correlation excitation, radiation, etc., and are applicable to the liquid environment of biological detection:

Experimental example

1. Electromagnetic wave near-field focusing detection of the chip of embodiment 1.

FIG. 10 is a gallium phosphide resonator chip provided by an embodiment of the present application. Among them, in Fig. 10, (a) top view of scanning electron microscope, (b) near-field focus distribution of electromagnetic waves under 459nm wavelength light excitation condition, (c) near-field focus distribution of electromagnetic wave under 584nm wavelength light excitation condition.

Figure 10(a) demonstrates that electron beam lithography and reactive ion etching can realize slits less than 30 nm wide in semiconductor disks. From the preliminary simulation calculation, as shown in Figure 10(b) and 10(c), placing the slit in the center of the disc under the non-pole mode (459nm, 584nm excitation wavelength) can effectively concentrate the electromagnetic field, the highest Values can go up to 120.

2. Detection of the electric field strength of the chip in embodiment 1.

Fig. 11 is a preliminary calculation and processing diagram of the gallium phosphide resonator chip provided by the embodiment of the present application, (a) scattering spectrum, (b) absorption spectrum, and (c) electromagnetic field enhancement performance of the resonator under different slit lengths. (d) Processed GaP resonator chips with different slit sizes.

Figure 11(a) and Figure 11(b) show the variation trend of the scattering and absorption spectra of the resonator with different slit lengths under preliminary simulation calculations. It can be seen that there are two scattering valleys in the resonator chip at the excitation wavelength, which corresponds to At the same time, it can be seen from the electric field intensity enhancement in Figure 11(c) that the electric field intensity in the resonant channel in the non-polar state is generally enhanced, and "double resonance enhancement" also appears at the excitation wavelength. Figure 11(d) shows the processed semiconductor resonator chips with different slit sizes.

3. Detect the electromagnetic field enhancement performance of the chips of Example 1 and Comparative Example 1. The result is shown in Figure 12.

FIG. 12 is a preliminary verification of the gallium phosphide resonant chip provided in the embodiment of the present application. (a) Comparison of absorption and scattering spectra of GaP resonators and ZMWs. (b) Electromagnetic field enhancement performance in the GaP resonator slot and ZMW hole.

Figure 12(a) shows the comparison of the absorption and scattering cross sections of the GaP resonator chip and the ZMW. It can be seen that the GaP resonator chip has lower absorption than the ZMW, and the GaP resonator has two There are two distinct scattering troughs, and these two troughs correspond to the resonant modes of the resonator. At the same time, according to the electromagnetic field enhancement performance in Figure 12(b), the gallium phosphide resonant chip has a strong electric field energy in the resonance mode, which is an order of magnitude higher than the electric field energy in the hole of the ZMW.

4. Detect the enhanced fluorescence quantum yields of the chips of Example 1 and Comparative Example 1. The result is shown in Figure 13.

Fig. 13 is the computational verification of the dielectric resonant chip provided in the embodiment and comparative example of the present application. (a) Comparison of enhanced fluorescence quantum yields of ZMW and GaP resonator chips. (b) Comparison of fluorescence radiation enhancement of ZMW and GaP resonator chips.

Figure 13(a) shows the comparison of the enhanced fluorescence quantum yield of ZMW and GaP resonator chips. Gallium phosphide resonators can produce higher fluorescence quantum yields in the visible light band, where the two spectra on the left are extracted from The information of 4 fluorescent wavelengths is summarized in the enhanced fluorescence quantum yield line on the right (the solid line represents the gallium phosphide resonant chip, and the dotted line represents the ZMW). Compared with the ZMW, the gallium phosphide resonant chip can produce an order of magnitude higher fluorescence quantum yield. Fig. 13(b) shows the contrast of the enhanced fluorescence radiation of ZMW and GaP resonator chips, and the GaP resonator can produce higher fluorescence radiation in the non-polar mode, in which 4 are extracted from the two spectra on the left The fluorescence wavelength information is summarized in the enhanced fluorescence radiation spectrum line on the right (the solid line represents the gallium phosphide resonator chip, and the dotted line represents the ZMW). Compared with the ZMW, the gallium phosphide resonator chip can produce an order of magnitude higher fluorescence radiation enhancement .

5. Calculation verification of the enhanced fluorescence of the chips of Example 1 and Comparative Example 1. The result is shown in Figure 14.

Fig. 14 Calculation verification of the enhanced fluorescence of the dielectric resonant chip provided in the examples and comparative examples of the present application. (a) Intrapore distribution of ZMW fluorescence radiation. (b) In-slit distribution of the fluorescence emission of a GaP resonator. (c) In-slit distribution of the fluorescence emission of a GaN resonator.

Figure 14(a) shows the distribution of fluorescence radiation in ZMW pores, the fluorescence intensity at 555nm/568nm radiation wavelength is higher than its intensity at 647nm/660nm radiation wavelength; Figure 14(b), Figure 14(c) The distribution of fluorescence radiation in the slits of gallium phosphide and gallium nitride resonators is shown respectively, and it is obvious that the enhancement effect of the dielectric resonator chip on fluorescence is better than that of the ZMW structure.

6. Preliminary verification of the enhanced excitation of the chips of Example 1 and Comparative Example 1. The result is shown in Figure 15.

Fig. 15 is a preliminary verification of the enhanced excitation of the dielectric waveguide resonant chip provided by the embodiment of the present application. (a) Schematic diagram of four excitation modes. (b) The electric field enhancement spectrum under the corresponding excitation mode. (c) The electric field distribution map under the corresponding excitation mode.

Figure 15(a) shows how the waveguide excites the resonant chip, including waveguide end-face coupling, embedded coupling, and side coupling (different slit orientations). According to the electric field enhancement results in Figure 15(b), the end-face coupling method Higher electric field enhancement can be produced at the excitation wavelength of 500nm-700nm. From the analysis of the electric field distribution in Figure 15(c), end-face coupling, embedded coupling and side coupling (the long axis of the slit is parallel to the waveguide) can better integrate The energy is confined in the slit channel of the resonator, which facilitates the on-chip excitation of the resonator chip.

7. Preliminary calculation verification of the enhanced quantum yield of the chips of Example 1 and Comparative Example 1. The result is shown in Figure 16.

Fig. 16 is a preliminary calculation and verification of the enhanced quantum yield of the dielectric waveguide resonant chip provided by the embodiment of the present application. (a) Schematic diagram of different units on the waveguide resonator chip, and (b) fluorescence quantum yield enhancement spectra when the initial quantum yield is 0.3 and (c) when the initial quantum yield is 0.003. (d) Schematic diagram of on-chip collection and radiation directionality.

Figure 16(a) shows the placement of the fluorescent source in different units of the waveguide resonator chip, including placement on a single chip substrate, placement in the resonant channel of the resonator, placement in the resonator with a substrate, and placement in the waveguide resonator chip, According to the quantum yield enhancement results in Figure 16(b) and 16(c), the existence of the resonator plays a key role in enhancing the quantum yield, and the waveguide resonator chip can produce more High fluorescence enhancement. From the radiation directivity analysis in Figure 16(d), the waveguide resonator chip can couple more fluorescence radiation energy into the waveguide due to the design of the on-chip waveguide, further realizing the on-chip collection of fluorescence signals.

The above descriptions are only preferred embodiments of the present application, and are not intended to limit the present application. For those skilled in the art, there may be various modifications and changes in the present application. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of this application shall be included within the protection scope of this application.

Claims

A resonant chip, characterized in that it comprises:

a dielectric layer, on which a resonant channel is arranged, and the resonant channel penetrates through the dielectric layer;

The dielectric layer includes a resonator and a transition dielectric layer;

The resonator is formed on the surface of the transition dielectric layer;

The resonance channel passes through the resonator and the transition dielectric layer in turn; the resonator is used to form a resonance response with the excitation light, and makes the electric field energy confined in the resonance channel; the transition dielectric layer is used for The passage of biomolecules is isolated, allowing biological reactions to occur only within the resonant channel.
The resonator chip according to claim 1, wherein the resonator is island-shaped.
The resonator chip according to claim 2, wherein the edge of the resonator forms a regular shape or a topological shape.
The resonant chip according to claim 1, wherein the resonant channel is in the shape of a slit, a regular hole or an irregular hole.
The resonant chip according to claim 4, wherein,

The maximum width of the narrowest part of the resonant channel is 20nm.
The resonant chip according to claim 4, wherein,

The total volume of the resonant channel is controlled at the level of single molecule detection.
The resonant chip according to any one of claims 1-6, wherein the dielectric layer is made of non-metallic material.
The resonator chip according to claim 7, wherein the materials of the resonator and the transition dielectric layer may be the same or different.
The resonant chip according to claim 1, wherein,

The resonant chip also includes:

Substrate;

A conductive layer formed on the surface of the substrate.
The resonant chip according to claim 9, wherein,

The conductive layer is made of electrode material for external voltage.
The resonant chip according to claim 9, wherein,

The resonant chip also includes:

an adhesion layer formed on the surface of the conductive layer; the dielectric layer is formed on the surface of the adhesion layer.
The resonant chip according to claim 11, characterized in that,

The adhesion layer is made of porous material.
The resonant chip according to claim 11, characterized in that,

The resonant chip includes a water seepage hole; the water seepage hole passes through the substrate and the conductive layer, and is in contact with the adhesion layer.
A resonant chip, characterized in that the resonant chip comprises:

Substrate;

a conductive layer on one side of the substrate;

an adhesion layer on one side of the conductive layer;

A dielectric layer located on one side of the adhesion layer, wherein a resonant slit is arranged on the dielectric layer, and the resonant slit penetrates through the dielectric layer.
The resonator chip according to claim 14, wherein the dielectric layer comprises a resonator and a transition dielectric layer;

The transition dielectric layer is located on one side of the adhesion layer, and the resonator is located on one side of the transition dielectric layer;

The resonant slit sequentially passes through the resonator and the transition dielectric layer.
The resonant chip according to claim 15, wherein the resonator is circular, the shape of the resonant slit is a strip-shaped rectangle, and the resonant slit is arranged at the center of the resonator.
The resonator chip according to claim 15, wherein the material for making the resonator and the transition dielectric layer includes gallium phosphide.
The resonant chip according to claim 15, wherein the dielectric layer further comprises a dielectric waveguide, and the dielectric waveguide is located on the transition dielectric layer.
The resonator chip according to claim 18, wherein the light beam emitted by the dielectric waveguide is horizontally incident on the resonator.
The resonator chip according to claim 18, wherein the light beam emitted by the dielectric waveguide is parallel to the resonator.
The resonant chip according to claim 14, wherein water seepage holes are further provided on the conductive layer and the substrate, and the positions of the water seepage holes correspond to the resonant slits.
A method for manufacturing a resonant chip, characterized in that the method comprises:

providing a substrate;

growing a conductive layer along one side of the substrate;

creating an adhesion layer along one side of the conductive layer;

A dielectric layer is fabricated along one side of the adhesion layer, wherein a resonant slit is arranged on the dielectric layer, and the resonant slit penetrates through the dielectric layer.
The method for manufacturing a resonant chip according to any one of claims 1-13, characterized in that the method comprises:

setting a resonant channel on the dielectric layer, so that the resonant channel penetrates through the dielectric layer;

The dielectric layer includes a resonator and a transition dielectric layer, so that the resonator is formed on the surface of the transition dielectric layer; the resonant channel passes through the resonator and the transition dielectric layer in sequence; the The resonator is used to form a resonance response with the excitation light, and to confine the electric field energy in the resonance channel; the transition dielectric layer is used to isolate biomolecules from passing through, so that biological reactions only react in the resonance channel.