CN107058082A - A kind of single-molecule DNA Non-Destructive Testing chip based near field optics - Google Patents
A kind of single-molecule DNA Non-Destructive Testing chip based near field optics Download PDFInfo
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- CN107058082A CN107058082A CN201710268352.9A CN201710268352A CN107058082A CN 107058082 A CN107058082 A CN 107058082A CN 201710268352 A CN201710268352 A CN 201710268352A CN 107058082 A CN107058082 A CN 107058082A
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Abstract
The invention discloses a kind of single-molecule DNA Non-Destructive Testing chip based near field optics, including silicon nitride board, the upper surface of base plate sets fluted, groove is from left to right successively by rectangle DNA molecular sample cell, tapered tunnel, narrow tunnel, gradually wide heading and the connection of rectangle waste liquid pool are constituted, graphene oxide membrane is equipped with narrow tunnel, the first and second Gold nanoparticle monofilms are respectively arranged with the left and right ends of graphene oxide membrane, electrode A is arranged in the DNA molecular sample cell of rectangle, electrode B is arranged in the waste liquid pool of rectangle, electrode is connected by wire with power supply.During sequencing, single strand dna passes through narrow tunnel under the collective effect of electric field force and graphene oxide with straight-chain, when by having the graphene capture zone detection zone of bridge skylight structure, Tip-Enhanced Raman Spectroscopy system can capture base Raman signal, and pass to computer.By analyzing and processing, the real-time detection of single strand dna sequence is realized.
Description
Technical field
The invention belongs to biomolecule detection technical field, and in particular to a kind of unimolecule based near field optics
DNA Non-Destructive Testing chips.
Background technology
DNA sequencing technology, as one of most important biomedical research means, to genomics and its related discipline
Development plays vital effect.And the lossless sequencing technologies of unimolecule are the latest development directions of current DNA sequencing technology,
Sequencing, long sequence output and low cost aspect have huge potentiality at a high speed.The DNA single-molecule sequencings technology that presently, there are is mostly
Based on nano-pore method, i.e., rub the biological electricity that produces by base and nano-pore to determine base sequence.But biological electricity is known
The specificity of other mode is not high, and anti-interference is poor, causes single base to recognize that error rate is high.In addition, existing single-molecule DNA
Also there is following shortcoming in sequencing technologies：(1) length is read smaller；(2) DNA fragments mosaicings error rate is high；(3) speed is slower.Cause
This, needs a kind of new and effective single-molecule DNA sequencing technology badly.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art to read length, high-accuracy, height there is provided a kind of high specific, height
The single-molecule DNA Non-Destructive Testing chip based near field optics of speed sequencing.
Technical scheme is summarized as follows：
A kind of single-molecule DNA Non-Destructive Testing chip based near field optics, including silicon nitride board 6, in silicon nitride
Upper surface of base plate sets fluted, and groove is from left to right successively by the DNA molecular sample cell 1, tapered tunnel 2, narrow tunnel of rectangle
3rd, gradually the connection of waste liquid pool 5 of wide heading 4 and rectangle is constituted, and (the bridge knot of graphene oxide membrane 7 is equipped with narrow tunnel 3
Structure), the first Gold nanoparticle monofilm 8 and the second golden nanometer particle are respectively arranged with the left and right ends of graphene oxide membrane 7
Monofilm 9 (skylight structure), electrode A is arranged in the DNA molecular sample cell 1 of rectangle, and electrode B is arranged on the waste liquid pool 5 of rectangle
Interior, electrode A and electrode B are connected by wire with power supply respectively.
Preferably, the width of the DNA molecular sample cell of rectangle is 100nm, and the width in narrow tunnel is 10nm, the width of waste liquid pool
Spend for 100nm.
The size in the gap 10 between the first Gold nanoparticle monofilm and the second Gold nanoparticle monofilm is 0.45-
0.65nm.Advantages of the present invention：
(1) present invention utilizes graphene oxide membrane, can be effectively by intermolecular π-π effects to the list of DNA molecular
Base is finely captured, and limits the randomness of its thermodynamics and kinetics, it is ensured that Tip-Enhanced Raman Spectroscopy system is near
100% capture rate collects single base information, and the response speed and precision of detecting system is greatly improved.
(2) present invention can efficiently control DNA motion to flow to and accurately input detection zone.
(3) graphene oxide and the local fields π-π of single base formation are acted on, and can be effectively improved single base Raman scattering and be cut
Face, so as to strengthen Raman signal；On this basis, the cloud density in local fields can greatly be strengthened by π-pi-electron effect,
Stronger TERS effects can be formed in plasma domain space.
(4) use graphene oxide for the saturating conductor of light, Gold nanoparticle monofilm is covered above, can be in effectively shielding needle point
While on the electric field influence of single base, the collection of near-field spectrum is not influenceed.
(5) because the universal resolution ratio of current Tip-Enhanced Raman Spectroscopy system is 20nm, the present invention is used with bridge
The graphene capture zone (i.e. graphene oxide membrane 7 is mounted on silicon nitride board across narrow tunnel 3) of structure is further to be lifted
The spatial resolution of Tip-Enhanced Raman Spectroscopy system；The capture zone covers graphene oxide using Gold nanoparticle monofilm
Film, reserves 0.45nm-0.65nm skylight (the first Gold nanoparticle monofilm and second in the way of AFM is manipulated
Gap between Gold nanoparticle monofilm) to match the space length of single base, and then by point of Tip-Enhanced Raman Spectroscopy
Resolution is promoted to 0.55nm or so, so as to recognize single base.
Brief description of the drawings
Fig. 1 is a kind of schematic front view of the single-molecule DNA Non-Destructive Testing chip based near field optics of the present invention.
Fig. 2 is a kind of schematic top plan view of the single-molecule DNA Non-Destructive Testing chip based near field optics of the present invention.
Embodiment
Below by accompanying drawing, the present invention is further illustrated.
A kind of single-molecule DNA Non-Destructive Testing chip (see Fig. 1, Fig. 2) based near field optics, including silicon nitride board
6, set fluted in silicon nitride board upper surface, groove is from left to right successively by the DNA molecular sample cell 1 of rectangle, tapered hole
Road 2, narrow tunnel 3, gradually wide heading 4 and rectangle waste liquid pool 5 connection composition, be equipped with graphene oxide membrane on narrow tunnel 3
7, the first Gold nanoparticle monofilm 8 and the second golden nanometer particle list are respectively arranged with the left and right ends of graphene oxide membrane 7
Tunic 9, electrode A is arranged in the DNA molecular sample cell 1 of rectangle, and electrode B is arranged in the waste liquid pool 5 of rectangle, electrode A and electricity
Pole B is connected by wire with power supply respectively.
Using focused ion beam (FIB) nanofabrication technique, by silicon nitride (Si3N4) substrate, width is processed for 100nm
Rectangle DNA molecular sample cell 1, tapered tunnel 2, width is 10nm narrow tunnel 3, and gradually wide heading 4 and width are 100nm's
Rectangle waste liquid pool 5, electrode A is arranged in DNA molecular sample cell 1, and electrode B is arranged in waste liquid pool 5, electrode A and electrode B difference
It is connected by wire with power supply, graphene oxide membrane 7 is carried on narrow tunnel, it is formed bridge structure on narrow tunnel
Capture zone, the opaque gold nano of first Gold nanoparticle monofilm 8 second is respectively arranged with the left and right ends of graphene oxide membrane 7
The size in the gap between particle monofilm 9, the two films is 0.55nm or 0.45nm or 0.65nm, with effective
With DNA base space length, and then the sun-roof composite construction of bridge is formed so that follow-up Tip-Enhanced Raman Spectroscopy is examined
Survey.
A kind of single-molecule DNA Non-Destructive Testing chip based near field optics of the present invention uses pinpoint enhanced Raman light
Spectra system is detected.
A kind of use of the single-molecule DNA Non-Destructive Testing chip based near field optics：
By concentration 1mol/L, pH value is is paved with 8.0 NaCl aqueous solution input groove, then by separated to be single-stranded
DNA molecular is put into the DNA molecular sample cell 1 of rectangle, then the chip of the present invention is placed on into Tip-Enhanced Raman Spectroscopy system
Sample stage on, and the probe of Tip-Enhanced Raman Spectroscopy system is placed on the first Gold nanoparticle monofilm 8 and the second Jenner
The top of graphene oxide membrane 7 between rice corpuscles monofilm 9；
Give electrode to be powered, by electrode to the chip with electric field force, due to graphene oxide and single strand dna π-
The collective effect of pi accumulation and electric field force, electronegative DNA molecular can keep single-chain state, by tapered tunnel 2, into narrow hole
Road 3.
When DNA molecular passes through the position of Tip-Enhanced Raman Spectroscopy system probe, system object lens focus on excitation beam
On probe tip top, the local enhanced field of generation can then excite the DNA sample base below needle point on silicon nitride board (6)
Raman signal.The Raman signal of local field is collected by the object lens of Tip-Enhanced Raman Spectroscopy system, and signal is introduced into needle point
Strengthen Raman spectrum system and carry out signal transacting.Tip-Enhanced Raman Spectroscopy system can record the Raman letter of now single base automatically
Number.DNA molecular can be under the driving of electric field force, into waste afterwards, and then completes the Non-Destructive Testing of single DNA molecules.
Experiment：Detect four kinds of bases in DNA
Experiment material：Artificial synthesized nucleotide fragments AGCTAGCT.
Instrument is set：Tip-Enhanced Raman Spectroscopy system, laser intensity settings are 100mW, and optical maser wavelength is 532nm, are received
The collection time is 0.1 millisecond.A kind of single-molecule DNA Non-Destructive Testing chip based near field optics of the present invention, the first Jenner
Gap (10) between rice corpuscles monofilm and the second Gold nanoparticle monofilm is 0.55nm.
Operating process：It is to be paved with the NaCl aqueous solution input groove that 1mol/L, pH value are 8.0 by concentration, this is artificial
The nucleotide fragments AGCTAGCT of synthesis is injected in the DNA molecular sample cell 1 of rectangle, then the chip of the present invention is placed on into needle point
On the sample stage for strengthening Raman spectrum system, the first Gold nanoparticle monofilm 8 and the second gold medal are found by AFM
The center of graphene oxide membrane 7 between nano particle single layer film 9.Give electrode be powered, by electrode to the chip with
Electric field force, electronegative DNA molecular can keep single-chain state, by tapered tunnel 2, into narrow tunnel 3.When electrode is initially powered up
Afterwards, that is, Tip-Enhanced Raman Spectroscopy signal acquisition is proceeded by, sequentially to obtain single base spectral signal, when all nucleosides
Acid enters after waste liquid pool, stop signal collection.
As a result：Table 1 is used after this artificial synthesized different base of four kinds of fragment of Tip-Enhanced Raman Spectroscopy system detectio
The obtained Raman spectrum spectral peak corresponding to base species.It can be seen that, the Raman signatures spectral peak of different bases has uniqueness
And not overlapping with other base features, the foundation that can be recognized as single base, with high accuracy.
The corresponding pinpoint enhanced Raman spectrum peak position of 1 four bases of table
Claims (3)
1. a kind of single-molecule DNA Non-Destructive Testing chip based near field optics, including silicon nitride board (6), in silicon nitride
Upper surface of base plate set it is fluted, it is characterized in that the groove from left to right successively by the DNA molecular sample cell (1) of rectangle, gradually
Narrow tunnel (2), waste liquid pool (5) the connection composition of narrow tunnel (3), gradually wide heading (4) and rectangle, are carried on narrow tunnel (3)
There is graphene oxide membrane (7), the first Gold nanoparticle monofilm is respectively arranged with the left and right ends of graphene oxide membrane (7)
(8) and the second Gold nanoparticle monofilm (9), electrode A is arranged in the DNA molecular sample cell (1) of rectangle, and electrode B is arranged on
In the waste liquid pool (5) of rectangle, electrode A and electrode B are connected by wire with power supply respectively.
2. a kind of single-molecule DNA Non-Destructive Testing chip based near field optics according to claim 1, it is characterized in that
The width of the DNA molecular sample cell of the rectangle is 100nm, and the width in the narrow tunnel is 10nm, the width of the waste liquid pool
For 100nm.
3. a kind of single-molecule DNA Non-Destructive Testing chip based near field optics according to claim 1, it is characterized in that
The size in the gap (10) between the first Gold nanoparticle monofilm and the second Gold nanoparticle monofilm is 0.45-0.65nm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710268352.9A CN107058082A (en) | 2017-04-22 | 2017-04-22 | A kind of single-molecule DNA Non-Destructive Testing chip based near field optics |
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| CN201710268352.9A CN107058082A (en) | 2017-04-22 | 2017-04-22 | A kind of single-molecule DNA Non-Destructive Testing chip based near field optics |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109652519A (en) * | 2018-12-20 | 2019-04-19 | 郑州大学 | A kind of atomic scale DNA sequencing method of logic-based judgement |
| CN109837207A (en) * | 2019-02-28 | 2019-06-04 | 中国科学院半导体研究所 | Gene sequencing chip and method |
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| CN103521780A (en) * | 2013-11-04 | 2014-01-22 | 中国农业科学院农业质量标准与检测技术研究所 | Preparation method and application of oxidized graphene load gold nanoparticle sol with surface-enhanced Raman spectrum activity |
| CN103890580A (en) * | 2011-09-28 | 2014-06-25 | 皇家飞利浦有限公司 | Apparatus for the processing of single molecules |
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2017
- 2017-04-22 CN CN201710268352.9A patent/CN107058082A/en active Pending
Patent Citations (3)
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| CN103890580A (en) * | 2011-09-28 | 2014-06-25 | 皇家飞利浦有限公司 | Apparatus for the processing of single molecules |
| CN102911861A (en) * | 2012-10-16 | 2013-02-06 | 沈阳市东陵区（浑南新区）卓科技术开发中心 | Nanoscale detection chip |
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| REGINA TREFFER ET A;.: "Distinction of nucleobases – a tip-enhanced Raman approach", 《BEILSTEIN J. NANOTECHNOL.》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109652519A (en) * | 2018-12-20 | 2019-04-19 | 郑州大学 | A kind of atomic scale DNA sequencing method of logic-based judgement |
| CN109652519B (en) * | 2018-12-20 | 2021-11-23 | 郑州大学 | Atomic scale DNA sequencing method based on logic judgment |
| CN109837207A (en) * | 2019-02-28 | 2019-06-04 | 中国科学院半导体研究所 | Gene sequencing chip and method |
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| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Chen Da Inventor after: Lin Jiahai Inventor before: Chen Da Inventor before: Zong Jing Inventor before: Li Qifeng |
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| TA01 | Transfer of patent application right | ||
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Effective date of registration: 20170925 Address after: 318000, room 11, No. 139, Zhongshan branch, Jiaojiang District, Zhejiang, Taizhou Applicant after: Zhejiang Da Da Technology Co., Ltd. Address before: 300000, No. 3699, Jinzhong Road, Dongli, 14-314, North China Hardware City G District, Tianjin Applicant before: TIANJIN ZHIQIAO DATA TECHNOLOGY CO., LTD. |
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Effective date of registration: 20181207 Address after: 300000 Jinzhong Highway 3699, Dongli District, Tianjin City, 14-314, G District, Hardware City, North China Applicant after: TIANJIN ZHIQIAO DATA TECHNOLOGY CO., LTD. Address before: Room 139, No. 11, Zhongshan Branch Road, Jiaojiang District, Taizhou City, Zhejiang Province Applicant before: Zhejiang Da Da Technology Co., Ltd. |
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Application publication date: 20170818 |