CN103105422B - Single-layer graphene nanopore structure for detection of base sequence and preparation method thereof - Google Patents
Single-layer graphene nanopore structure for detection of base sequence and preparation method thereof Download PDFInfo
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Abstract
The invention provides a single-layer graphene nanopore structure for the detection of a base sequence and a preparation method thereof. The preparation method comprises the following steps of shifting a graphene microchip onto the surface of a silicon oxide insulation layer supported by a semiconductor layer, preparing a metal micro electrode on the upper surfaces of two end surfaces of the graphene microchip, then preparing an insulation layer so as to cover the surfaces of the graphene microchip and the metal micro electrode to form a sandwich structure, finally realizing the penetration of the silicon oxide insulation layer, the graphene microchip and the insulation layer through releasing the semiconductor layer and preparing the nanopore. When basic group penetrates through the nanopore, a basic group electrical signal is identified and detected so as to realize the identification of the base sequence. The single-layer graphene nanopore structure has a simple preparation method, has low cost and small structure volume, has better extensibility by being compatible with a CMOS (complementary metal-oxide-semiconductor transistor) technology, and has broader application prospect in biomedical fields.
Description
Technical field
The invention belongs to microelectromechanical systems field, relate to a kind of single-layer graphene nano-pore structure and preparation method thereof, particularly relate to a kind of single-layer graphene nano-pore structure detecting for base sequence and preparation method thereof.
Background technology
Utilize biology sensor that microelectron-mechanical process technology makes to be widely applied in the field such as biological, medical.These sensors have cost low, can be mass, the advantage such as good stability, sensitivity height.When determinand is of a size of nanometer or Subnano-class, require sensor must have very high sensitivity, just can carry out effective recognition detection.Improving one of sensitivity effective ways is the size that reduces sensor detecting element, could effectively in time domain and spatial domain, detect determinand like this.Therefore, produce the sensor of nano-scale, can effectively improve the sensitivity of sensor, the quality factor of device is significantly improved.
In all multi-methods of current detection DNA base sequence, because electrode separation and DNA base size be not on same yardstick, cause detecting effect and be restricted.Xiaogan Liang, Stephen Y.Chou is at Nanogap Detector Inside Nanofluidic Channel for Fast Real-Time Label-Free DNA Analysis, 8,2008, in Nano Letters, describe the manufacturing technology of the nano-electrode detecting for DNA sequence dna in detail, but the minimum electrode spacing of preparing is 9nm, can not realize the identification of DNA base sequence, and point out that electrode separation size need to further reduce, just can make nano-electrode detect DNA base sequence becomes possibility.But the method that author adopts, is subject to the constraint of existing manufacturing technology, cannot continue to reduce the size of electrode.Makusu Tsutsui, Yuhui He, Masayuki Furuhashi etc. are at Transverse electric field dragging of DNA in a nanochannel, 5,2012, in Scientific reports mono-literary composition, in nanowires of gold, produce size at the nanochannel of 200nm * 50nm * 60nm, with this nanochannel, as nano electrode, detect DNA base sequence.In literary composition, detect λ-DNA by the signal of this nanochannel, but detect DeGrain.This is because the size of this nanochannel is larger, makes to be flooded by other signal for identifying the tunnelling current signal of DNA base sequence.So further reduce detecting electrode size, it is the important leverage of identification DNA base sequence.
Electrode separation is the determinative that impact detects DNA base sequence sensor.Because the thickness of single-layer graphene is only 0.34nm, in the same size with the base spacing of DNA molecular, therefore, by the ultra-thin characteristic advantage of Graphene, Graphene is made into nano-pore as nano-electrode, can effectively detect the base sequence of DNA molecular.Zuzanna S.Siwy and Matthew Davenport, at Graphene opens up to DNA, have commented the prospect that grapheme nano-pore detects DNA sequence dna in 5,2010, Nature Nanotechnology, mono-literary composition.The current research with grapheme nano-pore detection DNA sequence dna has been subjected to great concern, is all to utilize nano level grapheme nano-pore constraint DNA.When DNA to be measured passes through grapheme nano-pore, the variation of the occupy-place generation current signal of DNA in nano-pore, and the signal changing with this is identified DNA base sequence.Yet the signal that this method detects also can not identify DNA base sequence, only can detect the feature that DNA to be measured crosses grapheme nano-pore.
Summary of the invention
Technical matters: the shortcoming of prior art in view of the above, the object of the present invention is to provide a kind of single-layer graphene nano-pore structure detecting for base sequence and preparation method thereof, oversize, the low problem of sensitivity of nano-electrode when being used for solving base sequence and detecting, realize prior art and COMS technology compatible mutually simultaneously, can effectively reduce manufacturing process complexity, the problems such as rising of controlling cost.
Technical scheme: the single-layer graphene nano-pore structure detecting for base sequence of the present invention, comprise the mask substrate connecting successively from bottom to up, semiconductor layer, insulating layer of silicon oxide, electrode layer and insulation course, mask substrate middle part is provided with release window, in semiconductor layer, be provided with and be positioned at the etching groove that discharges window top and be communicated with it, in insulating layer of silicon oxide, be provided with the silicon oxide nano pore that is positioned at etching groove top the monox nanometer blind hole being communicated with it and is positioned at monox nanometer blind hole central authorities top, the metal microelectrode that electrode layer comprises Graphene microplate and is positioned at Graphene microplate two ends and is electrically connected to it, on Graphene microplate, be provided with the grapheme nano-pore that is positioned at silicon oxide nano pore top and is communicated with it, on insulation course, be provided with two external windows that are symmetrically distributed in metal microelectrode outer end edges top, external window is for being connected with extraneous electricity, on insulation course, be also provided with the etching window that is positioned at center and the insulation course nano-pore that is positioned at etching window center, insulation course nano-pore is positioned at grapheme nano-pore top and is communicated with it.
In the present invention, Graphene microplate is single-layer graphene, and the insulating layer of silicon oxide connecting successively from bottom to up, electrode layer and insulation course form sandwich structure.
In the present invention, the nano-pore of the connection that silicon oxide nano pore, grapheme nano-pore and insulation course nano-pore form.
In a preferred version of the present invention, the material of mask substrate is that material is monox or silicon nitride, and the material of semiconductor layer is silicon, germanium or germanium silicon.
The method of the above-mentioned single-layer graphene nano-pore structure detecting for base sequence of preparation of the present invention, comprises the following steps:
1) at the downside of semiconductor layer, produce mask substrate, the upside of semiconductor layer is produced insulating layer of silicon oxide;
2) in the central etching of mask substrate, make and discharge window;
3) single-layer graphene is transferred to the upper surface of insulating layer of silicon oxide, and by graphical treatment, obtained the Graphene microplate of regular shape, Graphene microplate is positioned at and discharges central authorities directly over window;
4) at Graphene microplate two ends, make respectively a metal microelectrode, two metal microelectrodes are realized electricity by Graphene microplate and are connected, and Graphene microplate and metal microelectrode have formed electrode layer;
5) make the insulation course of capping oxidation silicon insulation course and electrode layer upper surface, on etching insulation course, be positioned at the position of metal microelectrode outer end edges top, obtain the external window for being connected with extraneous electricity, on etching insulation course, be positioned at the position of Graphene microplate top, the thickness that reduces this position insulation course, obtains etching window;
6) discharge semiconductor layer, obtain etching groove and the unsettled unsettled structure of composite membrane of insulating layer of silicon oxide, electrode layer and insulation course above etching groove;
7) in insulating layer of silicon oxide central authorities, produce and be positioned at the monox nanometer blind hole being also communicated with it above etching groove;
8) in insulating layer of silicon oxide central authorities, produce and be positioned at the silicon oxide nano pore being also communicated with it above monox nanometer blind hole, on Graphene microplate, produce and be positioned at above silicon oxide nano pore and the grapheme nano-pore being communicated with it, etching window central authorities on insulation course produce and are positioned at above grapheme nano-pore and the insulation course nano-pore being communicated with it, and silicon oxide nano pore, grapheme nano-pore and insulation course nano-pore form the nano-pore of a connection.
Beneficial effect: the invention provides a kind of single-layer graphene nano-pore structure detecting for base sequence and preparation method thereof, compared with prior art, tool of the present invention has the following advantages:
1) electrode size is little, and accuracy of detection is high.Detect DNA molecular base sequence, while requiring DNA to pass through electrode, the size of electrode is consistent with base size as much as possible.Because the thickness of Graphene is 0.34nm, in the same size with DNA molecular base spacing, therefore, with grapheme nano-pore, as nano-electrode, can effectively detect the base sequence of DNA molecular.
2) utilize nanometer hole that DNA chain is stretching, improve the resolution of DNA base sequence.With respect to existing detection technique, nano-pore is effectively controlled the shape of DNA, makes DNA pass through nano-pore to be conducive to most the mode of identification, improves the resolution of base identification.
To sum up, single-layer graphene nano-pore structure that the base sequence that a kind of volume provided by the invention is little, technique is simple and cost is low detects and preparation method thereof, when electrode size is controlled, by grapheme nano-pore, realize detection, utilize metal microelectrode to realize the inside and outside electrical connection of device simultaneously, realize the conveying of detection signal.
Accompanying drawing explanation
Fig. 1 is the structural representation of substrate;
Fig. 2 is the structural representation of electrode layer;
Fig. 3 is the structural representation of insulation course;
Fig. 4 is the structural representation that semiconductor layer surface is produced mask substrate and insulating layer of silicon oxide;
Fig. 5 is that mask substrate central authorities etch the structural representation that discharges window;
Fig. 6 is the structural representation of Graphene microplate after graphical;
Fig. 7 is the structural representation of the external metal microelectrode of Graphene microplate;
Fig. 8 is for covering the structural representation after insulation course;
Fig. 9 for making the structural representation after etching window on insulation course;
Figure 10 for making the structural representation after external window on insulation course;
Figure 11 is for discharging the structural representation after semiconductor layer;
Figure 12 makes the structural representation after nanometer blind hole in insulating layer of silicon oxide central authorities;
Figure 13 is for making the grapheme nano-pore structural entity schematic diagram of the present invention after nano-pore.
In figure, have: mask substrate 1, semiconductor layer 2, insulating layer of silicon oxide 3, electrode layer 4, insulation course 5, discharge window 11, etching groove 21, nanometer blind hole 31, silicon oxide nano pore 32, Graphene microplate 41, metal microelectrode 42, grapheme nano-pore 43, external window 51, etching window 52, insulation course nano-pore 53.
Embodiment
Below in conjunction with Figure of description and specific embodiment, further illustrate the present invention, those skilled in the art can understand other advantages of the present invention and effect easily by the disclosed content of this instructions.The present invention can also be implemented or be applied by other different embodiment, and the every details in this instructions also can be based on different viewpoints and application, carries out various modifications or change not deviating under spirit of the present invention.
Refer to Fig. 4 to Figure 13, it should be noted that, the diagram providing in following specific embodiment only illustrates basic conception of the present invention in a schematic way, satisfy and only show with assembly relevant in the present invention in graphic but not component count, shape and size drafting while implementing according to reality, during its actual enforcement, kenel, quantity and the ratio of each assembly can be a kind of random change, and its assembly layout kenel also may be more complicated.
As shown in Fig. 4 to Figure 13, the single-layer graphene nano-pore structure that the present invention detects for base sequence, its method for making comprises the following steps:
Step 1), as shown in Figure 4, first produces mask substrate 1 at the downside of the semiconductor layer 2 as matrix, and the upside of semiconductor layer 2 is produced insulating layer of silicon oxide 3.In the present embodiment, semiconductor layer 2 is silicon, and insulating layer of silicon oxide 3 is monox, and the material of mask substrate 1 is monox, utilizes thermal oxidation technology at upside and the downside of semiconductor layer 2, to obtain mask substrate 1 and insulating layer of silicon oxide 3 simultaneously.
In step 2) in, as shown in Figure 5, utilize reactive ion etching process (Reactive-Ion Etching, RIE) etch mask substrate 1, in mask substrate 1, form one and discharge window 11.
In step 3), as shown in Figure 6, Graphene is transferred to insulating layer of silicon oxide 3 surfaces, and pass through graphical treatment, obtain the Graphene microplate 41 of regular shape, realize Graphene microplate 41 and be positioned at central authorities directly over the release window (11) that semiconductor discharge to finish to obtain.In the present embodiment, utilize photoetching and oxygen plasma etching, obtain the Graphene microplate 41 of regular shape.
In step 4), as shown in Figure 7, adopt sputter or evaporation, in insulating layer of silicon oxide 3 surfaces and Graphene microplate 41 surface deposition layer of metal, then by peeling off or lithographic method, form metal microelectrode 42, metal microelectrode 42 is realized electricity by Graphene microplate 41 and is connected.In the present embodiment, the material of metal microelectrode 41 is gold.
In step 5), as shown in Figure 8, make insulation course 5 total is covered.The insulation course of etching Graphene microplate 41 tops, reduces the thickness of insulating layer on Graphene microplate surface, is convenient to following process.Obtain etching window 52, as shown in Figure 9.The insulation course 5 of etching metal microelectrode 42 outer end edges tops, obtains external window 51, is used to form lead-in wire, realizes electrode and extraneous being electrically connected to, as shown in figure 10.It should be noted that, can first process and carve external window 51, then process fenetre mouth 52.In the present embodiment, first process etching window 52, then process external window 51.When making external window 51, the reserved very thin insulation course 5 of one deck in surface, the reserved insulation course 5 of external window 51, when step 6) discharges, can be corroded by alkaline solution, realizes with extraneous electricity and being connected.In the present embodiment, utilize hydrofluorite (Buffered Oxide Etch, the BOE) etching that photoetching and normal temperature cushioned to obtain external window 51 and fenetre mouth 52.
Step 6), puts into alkaline solution by total, utilizes step 2) the release window 11 that forms of etching discharges, remove semiconductor layer 2, obtain etching groove 21, and form insulation course 5 coverings of Hanging sectionally, the sandwich structure structure of composite membrane that electrode layer 4 and insulating layer of silicon oxide 3 form.Meanwhile, sharp alkaline solution release function, etches away the surperficial remaining insulation course of external window 51.Particularly, as shown in figure 11, in the present embodiment, the alkaline solution of removing semiconductor layer 2 is that concentration is 25% TMAH solution.
In step 7), insulating layer of silicon oxide 3 central authorities in etching groove 21 1 sides produce monox nanometer blind hole 31 as shown in figure 12. in the present embodiment, utilize focused ion beam (Focused Ion beam, FIB) etching oxidation silicon insulation course 3, at insulating layer of silicon oxide 3, form nanometer blind hole 31.
Step 8), as shown in figure 13, in monox nanometer blind hole, 31 central authorities make silicon oxide nano pore 32, and on Graphene microplate 41 and insulation course 5, produce successively grapheme nano-pore 43 and insulation course nano-pore 53; Realize the connection of silicon oxide nano pore 32, grapheme nano-pore 43 and insulation course nano-pore 53.In the present embodiment one, utilize transmission electron microscope (Transmission electron microscope, TEM) in nanometer blind hole 31 central authorities, produce silicon oxide nano pore 32, then process silicon oxide nano pore 32 place's processing graphite alkene nano-pores 43 and the nano-pore 53 layer by layer that insulate, form a nanometer through hole.
In sum, single-layer graphene nano-pore structure detecting for base sequence provided by the invention and preparation method thereof, has solved conventional fabrication processes and cannot reach the electrode problems of producing single Nano grade.This manufacture craft is simple, and cost is low and have advantages of that structural volume is little, and meanwhile, the present invention and CMOS process compatible make it have good extendability and wider usable range.So the present invention has effectively overcome various shortcoming of the prior art and tool high industrial utilization.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all can, under spirit of the present invention and category, modify or change above-described embodiment.Therefore, such as in affiliated technical field, have and conventionally know that the knowledgeable, not departing from all equivalence modifications that complete under disclosed spirit and technological thought or changing, must be contained by claim of the present invention.
Claims (4)
1. the single-layer graphene nano-pore structure detecting for base sequence, it is characterized in that, this structure comprises the mask substrate (1) connecting successively from bottom to up, semiconductor layer (2), insulating layer of silicon oxide (3), electrode layer (4) and insulation course (5), described mask substrate (1) middle part is provided with and discharges window (11), in described semiconductor layer (2), be provided with and be positioned at the etching groove (21) that discharges window (11) top and be communicated with it, in described insulating layer of silicon oxide (3), be provided with the silicon oxide nano pore (32) that is positioned at etching groove (21) top the monox nanometer blind hole (31) being communicated with it and is positioned at described monox nanometer blind hole (31) central authorities top, the metal microelectrode (42) that described electrode layer (4) comprises Graphene microplate (41) and is positioned at described Graphene microplate (41) two ends and is electrically connected to it, on described Graphene microplate (41), be provided with the grapheme nano-pore (43) that is positioned at silicon oxide nano pore (32) top and is communicated with it, on described insulation course (5), be provided with two the external windows (51) that are symmetrically distributed in metal microelectrode (42) outer end edges top, described external window (51) is for being connected with extraneous electricity, on insulation course (5), be also provided with the insulation course nano-pore (53) that is positioned at the etching window (52) at center and is positioned at described etching window (52) center, described insulation course nano-pore (53) is positioned at grapheme nano-pore (43) top and is communicated with it,
Described Graphene microplate (41) is single-layer graphene, and the insulating layer of silicon oxide (3) connecting successively from bottom to up, electrode layer (4) and insulation course (5) form sandwich structure.
2. the single-layer graphene nano-pore structure detecting for base sequence according to claim 1, it is characterized in that the nano-pore of the connection that described silicon oxide nano pore (32), grapheme nano-pore (43) and insulation course nano-pore (53) form.
3. the single-layer graphene nano-pore structure detecting for base sequence according to claim 1, is characterized in that: the material of described mask substrate (1) is monox or silicon nitride, and the material of described semiconductor layer (2) is silicon, germanium or germanium silicon.
4. a method of preparing the single-layer graphene nano-pore structure detecting for base sequence claimed in claim 1, is characterized in that, the method comprises the following steps:
1) at the downside of semiconductor layer (2), produce mask substrate (1), the upside of semiconductor layer (2) is produced insulating layer of silicon oxide (3);
2) in the central etching of described mask substrate (1), make and discharge window (11);
3) single-layer graphene is transferred to the upper surface of insulating layer of silicon oxide (3), and by graphical treatment, obtained the Graphene microplate (41) of regular shape, described Graphene microplate (41) is positioned at and discharges central authorities directly over window (11);
4) at described Graphene microplate (41) two ends, make respectively a metal microelectrode (42), described two metal microelectrodes (42) are realized electricity by Graphene microplate (41) and are connected, and Graphene microplate (41) and metal microelectrode (42) have formed electrode layer (4);
5) make the insulation course (5) that covers described insulating layer of silicon oxide (3) and electrode layer (4) upper surface, on etching insulation course (5), be positioned at the position of metal microelectrode (42) outer end edges top, obtain the external window (51) for being connected with extraneous electricity, on etching insulation course (5), be positioned at the position of Graphene microplate (41) top, reduce the thickness of this position insulation course, obtain etching window (52);
6) discharge described semiconductor layer (2), obtain etching groove (21) and unsettled insulating layer of silicon oxide (3), electrode layer (4) and the unsettled structure of composite membrane of insulation course (5) in etching groove (21) top;
7) in described insulating layer of silicon oxide (3) central authorities, produce the monox nanometer blind hole (31) that is positioned at etching groove (21) top and is communicated with it;
8) in described insulating layer of silicon oxide (3) central authorities, produce the silicon oxide nano pore (32) that is positioned at monox nanometer blind hole (31) top and is communicated with it, on Graphene microplate (41), produce the grapheme nano-pore (43) that is positioned at silicon oxide nano pore (32) top and is communicated with it, etching window (52) central authorities on insulation course (5) produce the insulation course nano-pore (53) that is positioned at grapheme nano-pore (43) top and is communicated with it, and silicon oxide nano pore (32), grapheme nano-pore (43) and insulation course nano-pore (53) form the nano-pore of a connection.
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| CN105776127B (en) * | 2016-04-22 | 2017-05-03 | 东南大学 | Method for manufacturing double-layer SiN nanopore structure for DNA base sequence detection |
| CN109182484B (en) * | 2018-09-07 | 2022-05-03 | 苏州罗岛纳米科技有限公司 | Nanopore sandwich structure for DNA base sequence detection and manufacturing method thereof |
| CN109261230B (en) * | 2018-09-30 | 2020-05-08 | 东南大学 | Monomolecular controllable output device of light-operated nanopore and use method thereof |
| CN109633154B (en) * | 2018-11-14 | 2022-02-08 | 广东工业大学 | Novel solid-state nanopore structure and manufacturing method thereof |
| CN109972087B (en) * | 2019-04-15 | 2020-01-07 | 华东师范大学 | Preparation method of microelectrode deposition mask |
| CN117203351A (en) * | 2021-08-31 | 2023-12-08 | 深圳华大生命科学研究院 | Microporous structure, preparation method and chip |
| CN114235037A (en) * | 2021-12-06 | 2022-03-25 | 中国科学院物理研究所 | High-aspect-ratio nano-gap multi-physical-field coupling detection platform and detection method |
| CN114715840B (en) * | 2022-04-11 | 2023-09-05 | 清华大学 | Differential suspension single-layer graphene nanopore sensor and preparation method and application thereof |
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| CN102445480B (en) * | 2011-09-23 | 2013-09-18 | 东南大学 | Method for preparing nano-gap electrodes on surface of nano-pore and in nano-pore |
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