CN1986383A - Single nano pore making process in single particle beam etched film - Google Patents
Single nano pore making process in single particle beam etched film Download PDFInfo
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- CN1986383A CN1986383A CN 200610147232 CN200610147232A CN1986383A CN 1986383 A CN1986383 A CN 1986383A CN 200610147232 CN200610147232 CN 200610147232 CN 200610147232 A CN200610147232 A CN 200610147232A CN 1986383 A CN1986383 A CN 1986383A
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Abstract
The single nanometer pore making process in single particle beam etched film belongs to the field of nanometer technology. The making process includes: setting insulating solid film on one horizontal rotating carrier with particle detecting unit and etching with single particle beam in the incident angle of 9.6-90 deg to penetrate the film, shifting the film relatively to the single particle beam and etching for the next time, and so on until the film is rotated by one turn to obtain truncated cone shaped, double turned truncated cone shaped or cylindrical single nanometer hole. The process can make single nanometer hole in the diameter of 1 nm level and depth up to micron level, and the single nanometer hole is suitable for distinguishing the base component of single strand DNA or RNA molecule and detecting the size of double strand DNA molecule. The present invention provides key device for nucleic acid sequencing.
Description
Technical field
The present invention relates to a kind of preparation method of nano-pore, particularly a kind of single nano pore making process of single-particle beam etched film.Belong to field of nanometer technology.
Background technology
Exist natural protein hole on the biological protoplast membrane, can carry out in the cell and extracellular ion-exchange, can detect the ion mobility status with patch-clamp, infect two adipose membranes with AH and form a kind of anisopleural protein nano hole, also can be used for the detection of ion stream, but more be the dynamics research that is used for nucleic acid and single stranded deoxyribonucleic acid (general designation nucleic acid) perforation, detect the size of nucleic acid molecule fragment, measure the potentiality of their base sequences in addition, but because this nano-pore is the hole that protein forms on two adipose membranes, aging easily, can not tolerate higher voltage, it is bigger that permeability is influenced by pH and salinity, also may exist and nucleic acid mutual effect site, under effective electric-field intensity, nucleic acid is too fast by the speed of nano-pore, the resolution ratio that has exceeded current patch-clamp, though the punching rate of nucleic acid has been reduced to the 3nt/ microsecond by regulating electrophoresis liquid component etc., but still exceeded 3 orders of magnitude of instrumental resolution, these all make the nano-pore order-checking become complicated.In order to overcome the natural defect that the albumen hole exists, carry out methods such as chemical etching, thin polymer film bundle hole and the interior plating of low-density commercial membranes again after people bore a hole with ion beam, electron beam lithography or sputter, heavy ion and prepare solid film, but the three dimension scale of nano-pore is uncontrollable, very irregular, so how to make the controlled nano-pore of yardstick, become to improve the key that detects charged molecule signal of telecommunication accuracy.But adopt in the past the solid nano hole nucleic acid chains is directly checked order research in, serious problems of being ignored by people, be that adjacent base spacing has only 7 on the single-chain nucleic acid, also be that existing solid nano hole hole depth is too big, generally in μ m level, can hold a plurality of bases simultaneously, like this, will cause obscuring of the signal of telecommunication and can't measure the actual sequence of base in the nucleic acid molecules.
Find through literature search prior art, people such as Golovchenko are at " Nature " (" Nature Journal ", calendar year 2001 the 412nd is rolled up the 166-169 page or leaf) delivered the paper that is entitled as " Ion-beam sculpting at nanometrelength scales " (" particle beams is carved ") under the long yardstick of nanometer, propose with Si
3N
4Substrate is a material, uses Ar
+The hole of the wide 1.8nm of outlet is made in particle beams sputter, the shape in hole is irregular, utilize bore 5nm, 500bp double-stranded DNA perforation dynamics is detected in the hole of the about 10nm of the degree of depth, find that its aperture time is all in 5 milliseconds, be that its perforation rate is 100bp/ Bo second, and applied for U.S. Patent number 7118657 according to this, patent is called the patent of " Pulsed ion beam control of solid state features " (" the pulse particle beams control of solid shape "), utilize this patented technology, though can obtain the element of nano-pore diameter at 5nm, but the shape of nano-pore is very irregular, at first be that the aperture is not circular, so when carrying out the double-stranded DNA length detection, the dna fragmentation of equal length does not have the identical aperture time, secondly be that the degree of depth in hole is uncontrollable, the nano-pore degree of depth of producing is at 10nm, exceed single-chain DNA base spacing 0.7nm, that is to say that Kong Zhongke holds about 15 bases simultaneously, therefore can't be used for the nano-pore order-checking of single strand dna.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, a kind of single nano pore making process of single-particle beam etched film is proposed, achieve the preparation of the controlled single nano-pore of two dimension (diameter and hole depth), that prepared single nano-pore is circular, aperture is controlled 1 nm level, hole depth is controlled to μ m level from the level, is suitable for detecting the size of nucleic acid molecules or differentiating its base component.
The present invention is achieved by the following technical solutions, and the inventive method is specially:
The solid insulation film of thickness at 2-5 μ m placed on the carrier that can horizontally rotate that has device for detecting particles, the above-mentioned solid insulation film of single-particle beam (charged particle beam or neutron beam) etching with beam cross section diameter 0.5-1nm, the single-particle beam incidence angle is more than or equal to 9.6 °, less than 90 °, and single-particle beam is in this film bottom, the top, or the somewhere intersects with solid insulation film rotating shaft between the two, after treating that single-particle beam penetrates this film fully, the particle monitor feedback of solid insulation film bottom makes the carrier rotation once, make the solid insulation film by the place of penetrating and single-particle beam 1 dust that relatively moves, etching again, so repeatedly, rotate a circle until the solid insulation film, obtain the about 1-3nm of minimum diameter, the nano-pore edge thickness is less than truncated cone-shaped or two reverse frustoconic list nano-pore of the spacing (0.7nm) of two adjacent bases in the nucleic acid chains, when the single-particle beam incidence angle is 90 °, etching obtains the tubular nano-pore of diameter at about 1nm until penetrating the solid insulation film continuously.
Described solid insulation film is silicon chip, mica sheet, diamond thin or quartz glass film.
Described single nano-pore, its shape depends on the incidence angle of single-particle beam and the crossover location of single-particle beam and solid insulation film rotating shaft, when the single-particle beam incidence angle more than or equal to 9.6 °, less than 90 °, the crossover location of single-particle beam and solid insulation film rotating shaft is when solid insulation film top or bottom, single nano-pore is a truncated cone-shaped, the crossover location of single-particle beam and solid insulation film rotating shaft is when the asking of solid insulation film upper and lower surfaces, single nano-pore is that (crossover location is two reverse frustoconics of symmetry to two reverse frustoconics when solid insulation film mid-height place; During other positions between solid insulation film upper and lower surfaces of crossover location, be asymmetric pair of reverse frustoconic); When the single-particle beam incidence angle was 90 °, single nano-pore was a cylindrical shape.
Truncated cone-shaped and various pairs of reverse frustoconic list nano-pores that the inventive method is made, the nano-pore inward flange degree of depth is less than 0.7nm, combine with electrophoresis tank and patch-clamp, can be accurately the adjacent base of single stranded DNA and RNA be detected respectively one by one, having overcome nano-pore in the past crosses deeply, holds a plurality of bases simultaneously and can't differentiate the shortcoming of base signal one by one, for the nucleic acid sequencing based on nano-pore provides Primary Component, with such device order-checking, be expected to realize ultrahigh speed, low cost, high accuracy, unnotched gene order-checking, satisfy multiple demand quick order-checking.Utilize the inventive method, also can obtain the single nano-pore of tubular, compare with the nano-pore that prior art is made, the degree of depth in its hole and controllable diameter, regular shape, inner bright and clean, can increase substantially signal to noise ratio, thereby can detect the size of double-stranded DNA more accurately.
The specific embodiment
Embodiment 1
The silicon chip of thickness 2 μ m is placed on the carrier that can horizontally rotate that has ion detection device, single-particle beam with beam cross section diameter 1nm, with 60 degree incidence angle etching silicon chips, the rotating shaft with silicon chip intersects in the silicon chip bottom to make single-particle beam, after treating that silicon chip is penetrated fully, the particle monitor feedback of silicon chip bottom makes the carrier rotation once, make silicon chip by the place of penetrating and single-particle beam 1 that relatively moves, silicon chip and particle beams sight relative position rotation 1nm, etching so repeatedly, rotates a circle until silicon chip again, obtain the about 1.2nm of minimum diameter, nano-pore inward flange thickness is at single nano-pore of 0.5nm, and this list nano-pore is a truncated cone-shaped.
There are the rectangle of the about 10 μ m apertures of a diameter or circular solid phase support thing in the silicon chip center of being fixed to that to contain single nano-pore, and (thickness is in the mm level, the length of side or diameter are in the cm level) go up and will seal around the silicon chip, nano-pore is aimed at holder aperture center, obtains size and is suitable for manual nano-component (mm-cm level).
Effect: make the single nano-pore of truncated cone-shaped with the single-particle beam etching, the diameter of single nano-pore can be controlled at 1.2nm, nano-pore inward flange THICKNESS CONTROL is at 0.5nm.
Embodiment 2
The mica sheet of thickness 5 μ m is placed on the carrier that can horizontally rotate that has device for detecting particles, single-particle beam with beam cross section diameter 1nm, with 45 degree incidence angle etching mica sheets, the particle beams is intersected with rotating shaft at half place of mica sheet thickness, after treating that mica sheet is penetrated fully, the particle monitor feedback of mica sheet bottom makes the mica sheet on the carrier be rotated 1 by the place of penetrating with particle beams sight relative position, etching again, so repeatedly, rotate a circle until mica sheet, obtain the about 1.5nm of minimum diameter, single nano-pore of the about 0.5nm of nano-pore edge thickness, this list nano-pore are two reverse frustoconics of symmetry.
There are the rectangle of a diameter 10 μ m apertures or circular solid phase support thing in the mica sheet center of being fixed to that to contain single nano-pore, and (thickness is in the mm level, the length of side or diameter are in the cm level) on, nano-pore is aimed at holder aperture center, and, obtain size and be suitable for manual nano-component (mm-cm level) sealing around the mica sheet.
Effect: make two reverse frustoconic list nano-pores with the single-particle beam etching, the diameter of single nano-pore can be controlled at 1.5nm, nano-pore inward flange THICKNESS CONTROL is at 0.5nm.
Embodiment 3
The diamond thin of thickness 2.5 μ m is placed on the carrier that can horizontally rotate that has device for detecting particles, single-particle beam with beam cross section diameter 1nm, with 90 degree incidence angle etching diamond films, penetrate fully until film, obtain the single nano-pore of tubular of the about 1nm of diameter, the nanometer hole depth is 2.5 μ m.
(thickness is in the mm level to prepare the rectangle that there is μ m level aperture at a center or circular solid phase support thing, the length of side or diameter are in the cm level), with the center of nano-pore alignment apertures on the diamond thin, sealing around the diamond thin obtains size and is suitable for manual nano-component.
Effect: make the single nano-pore of tubular with the single-particle beam etching, the diameter of single nano-pore can be controlled at about 1nm, the thickness of diamond thin is hole depth, i.e. 2.5 μ m.
Embodiment 4
The quartz glass film of thickness 5 μ m is placed on the carrier that can horizontally rotate that has ion detection device, single-particle beam with beam cross section diameter 0.5nm, with 9.6 degree incidence angle etching quartz glass films, the particle beams 2 μ m places on distance quartz glass film are intersected with rotating shaft, after treating that the quartz glass film is penetrated fully, the particle monitor feedback of quartz glass film bottom makes the carrier rotation once, make the quartz glass film by the place of penetrating and single-particle beam 1 that relatively moves, quartz glass film and particle beams sight relative position rotation 0.5nm, etching again, so repeatedly, rotate a circle until the quartz glass film, obtain the about 3nm of minimum diameter, single nano-pore of the about 0.5nm of nano-pore inward flange thickness, this list nano-pore are asymmetric pair of reverse frustoconic.
There are the rectangle of the about 10 μ m apertures of a diameter or circular solid phase support thing in the quartz glass film center of being fixed to that to contain single nano-pore, and (thickness is in the mm level, the length of side or diameter are in the cm level) go up and will seal around the quartz glass film, nano-pore is aimed at holder aperture center, obtains size and is suitable for manual nano-component (mm-cm level).
Effect: make the single nano-pore of truncated cone-shaped with the single-particle beam etching, the diameter of single nano-pore can be controlled at 3nm, nano-pore inner edge THICKNESS CONTROL is at 0.5nm.
Embodiment 5
The silicon chip of thickness 2 μ m is placed on the carrier that can horizontally rotate that has ion detection device, single-particle beam with beam cross section diameter 0.5nm, with 30 degree incidence angle etching silicon chips, the rotating shaft with silicon chip intersects at the silicon chip top to make single-particle beam, after treating that silicon chip penetrates fully, the particle monitor feedback of silicon chip bottom makes the carrier rotation once, make silicon chip by the place of penetrating and single-particle beam 1 that relatively moves, silicon chip and particle beams sight relative position rotation 1nm, etching so repeatedly, rotates a circle until silicon chip again, obtain minimum diameter 1nm, nano-pore inward flange thickness is at single nano-pore of 0.5nm, and this list nano-pore is a truncated cone-shaped.
There are the rectangle of the about 10 μ m apertures of a diameter or circular solid phase support thing in the silicon chip center of being fixed to that to contain single nano-pore, and (thickness is in the mm level, the length of side or diameter are in the cm level) go up and will seal around the silicon chip, nano-pore is aimed at holder aperture center, obtains size and is suitable for manual nano-component (mm-cm level).
Effect: make the single nano-pore of truncated cone-shaped with the single-particle beam etching, the diameter of single nano-pore can be controlled at 1nm, nano-pore inward flange THICKNESS CONTROL is at 0.5nm.
Claims (8)
1, a kind of single nano pore making process of single-particle beam etched film, it is characterized in that, described method is specially: the solid insulation film is placed on the carrier that can horizontally rotate that has device for detecting particles, with single-particle beam etching solid insulation film, the single-particle beam incidence angle more than or equal to 9.6 ° smaller or equal to 90 °, and single-particle beam is in this film bottom, the top, or intersect with solid insulation film rotating shaft between the two, after treating that single-particle beam penetrates this film fully, the particle monitor feedback of solid insulation film bottom makes the carrier rotation once, make the solid insulation film by the place of penetrating and single-particle beam 1 that relatively moves, etching again, so repeatedly, rotate a circle until the solid insulation film, obtain single nano-pore.
2, single nano pore making process of single-particle beam etched film according to claim 1 is characterized in that, described solid insulation film, its thickness are 2-5 μ m.
3, single nano pore making process of single-particle beam etched film according to claim 1 and 2 is characterized in that, described solid insulation film is silicon chip, mica sheet, diamond thin or quartz glass film.
4, single nano pore making process of single-particle beam etched film according to claim 1 is characterized in that, described single-particle beam is the single-particle beam of beam cross section diameter at 0.5-1nm.
5, single nano pore making process of single-particle beam etched film according to claim 1 is characterized in that, described single nano-pore is the nano-pore of diameter 1-3nm, bore edges thickness＜0.7nm, truncated cone-shaped or two reverse frustoconics.
6, single nano pore making process of single-particle beam etched film according to claim 1 is characterized in that, described single nano-pore is the tubular nano-pore of diameter 1nm.
7, single nano pore making process according to claim 1 or 5 or 6 described single-particle beam etched films, it is characterized in that, described single nano-pore, its shape depends on the incidence angle of single-particle beam and the crossover location of single-particle beam and solid insulation film rotating shaft, when the incidence angle of single-particle beam more than or equal to 9.6 ° less than 90 °, the crossover location of single-particle beam and solid insulation film rotating shaft is when solid insulation film top or bottom, single nano-pore is a truncated cone-shaped, the crossover location of single-particle beam and solid insulation film rotating shaft is between solid insulation film upper and lower surfaces the time, and single nano-pore is two reverse frustoconics; When the single-particle beam incidence angle was 90 °, single nano-pore was a tubular.
8, single nano pore making process of single-particle beam etched film according to claim 1 or 5, it is characterized in that, described single nano-pore, when the single-particle beam incidence angle more than or equal to 9.6 °, smaller or equal to 90 °, single-particle beam and solid insulation film rotating shaft crossover location are when solid insulation film mid-height place, two reverse frustoconics for symmetry, during other position between solid insulation film upper and lower surfaces of single-particle beam and solid insulation film rotating shaft crossover location, be asymmetric pair of reverse frustoconic.
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| CN 200610147232 CN1986383A (en) | 2006-12-14 | 2006-12-14 | Single nano pore making process in single particle beam etched film |
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| CN 200610147232 CN1986383A (en) | 2006-12-14 | 2006-12-14 | Single nano pore making process in single particle beam etched film |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105883838A (en) * | 2016-03-31 | 2016-08-24 | 北京大学 | Single-layer mica sheet and preparation method and application of nanopore electronic device thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105883838A (en) * | 2016-03-31 | 2016-08-24 | 北京大学 | Single-layer mica sheet and preparation method and application of nanopore electronic device thereof |
| CN105883838B (en) * | 2016-03-31 | 2018-08-21 | 北京大学 | The preparation method and application of single layer mica sheet and its nano-pore electronic device |
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Open date: 20070627 |