CN108226249A - Disposable nanometer aperture biosensor and preparation method thereof - Google Patents
Disposable nanometer aperture biosensor and preparation method thereof Download PDFInfo
- Publication number
- CN108226249A CN108226249A CN201810019657.0A CN201810019657A CN108226249A CN 108226249 A CN108226249 A CN 108226249A CN 201810019657 A CN201810019657 A CN 201810019657A CN 108226249 A CN108226249 A CN 108226249A
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- disposable
- micropore
- nanometer aperture
- head cover
- nano
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
Abstract
The invention discloses a kind of disposable nanometer aperture biosensors, include substrate, electrode layer and the head cover of stacked above one another setting, and head cover is provided with a diameter of 5 200 μm of micropore, and the micropore is used to inject nano-pore and phospholipid membrane.The nano-pore biosensor structure is simple, detection is convenient, of low cost, instant it can add adding in reagent section, ensure the stability of nano-pore and phospholipid membrane itself, it is particularly suitable for disposable occasion, it avoids and reuses sensor and lead to interfering with each other for testing result between patient, improve the accuracy of testing result.The production method for also disclosing the biosensor, process steps are simple, easily operated, in high volume can quickly produce, greatly reduce the production cost of nanometer aperture biosensor, be suitble to mass industrial production and wide popularization and application.
Description
Technical field
The invention belongs to technical field of biological, are related to a kind of biosensor and preparation method thereof, specifically relate to
And a kind of disposable nanometer aperture biosensor and preparation method thereof.
Background technology
Highly sensitive, highly selective quick sensing analysis around biomolecule was studied much and soon in recent years
Speed development, researcher combine a variety of nanometer technologies, using the advanced nano material for preparing and obtaining, have developed a series of electrifications
Biosensors, wherein nanometer aperture biosensor are one of which important biomolecule detection sensors.
Nano-pore refers generally to pore passage structure of the aperture in 1-10nm, pore size and single biological macro molecule size phase
When usual nano-pore is placed into organic phospholipid film, is analysed to sample (such as serum) and is separated in two parts, when voltage is applied to
During nano-pore both sides, charged ion and molecule in sample can pass through nano-pore and form electric current, and the current signal of generation can be by spy
It is fixed to acquire and analyzed together.When large biological molecule passes through nano-pore, due to the difference of the properties such as its size and charge, meeting
The current signature of specificity is generated, so as to analyze big point of biology in sample according to the current signal that instrument acquires
The information such as structure, type and the concentration of son.
Nano-pore includes two major class of solid nano hole and biological nano hole, and the nano-pore in being normally applied is biological nano
Hole, biological nano hole is with supporting its artificial phospholipid membrane and the stent of phospholipid membrane being supported to synthesize nano-pore bio-sensing
Device, nanometer aperture biosensor and nano-pore technology are widely used in the qualitative and quantitative analysis of large biological molecule, as DNA is surveyed
Sequence, tumor marker analyte detection and environment detection of heavy metal ion etc., nano-pore technology are that third generation gene popular in the world is surveyed
Sequence technology due to its huge applications potentiality in the fields such as biology, chemistry, medicine, food and environment, has developed into one
The brand-new research field of the multi-crossed disciplines such as biology, chemistry, medicine, fluid, electronics, material, machinery.
Diagnosis, micro-fluidic/to receive flow control and Molecular Detection field in vitro, nano-pore product has high sensitivity, operation letter
The plurality of advantages such as singly, detection speed is fast, equipment cost is low, the core of nano-pore product is a nanometer aperture biosensor, but existing
Some nanometer aperture biosensors are mostly the product reused, and reuse sensor and easily lead to phase between different patients
Mutually interference influences the accuracy of testing result, on the market there is also the nanometer aperture biosensor of some medical disposable material forms,
But its is complicated for operation, with high costs, up to hundreds of dollars, this high cost limits the extensive use of nano-pore technology.
Invention content
For this purpose, the technical problems to be solved by the invention are that Conventional nano aperture biosensor is reused, detection knot
Fruit is not accurate enough, and disposable sensor is complicated for operation, with high costs, can not extensive use, so as to propose one kind can quickly detect,
Disposable nanometer aperture biosensor of low cost and preparation method thereof.
In order to solve the above technical problems, the technical scheme is that：
The present invention provides a kind of disposable nanometer aperture biosensor, includes substrate, the electrode of stacked above one another setting
Layer and head cover, the head cover are provided with a diameter of 5-200 μm of the micropore for injecting nano-pore and phospholipid membrane.
Preferably, the head cover has the sample restricted area of a hollow out, an insulation is provided in the sample restricted area
Covering, the insulation cover surface are less than the surface of sample restricted area side wall, and the micropore is opened in the insulation
Covering.
Preferably, the substrate and the head cover are plastic layer, the electrode is to be set to the substrate surface
Silver electrode layer.
Preferably, the silver electrode layer is made of a pair of of silver electrode, the insulation cover is covered in the silver
Electrode surface, the micropore in the insulation cover are arranged at the top of silver electrode, and the silver electrode is made only to be exposed by the micropore.
Preferably, the size of the disposable nanometer aperture biosensor is not less than 5mm*5mm, the sample restricted area
Range diameter is not less than 1.5mm.
Preferably, the micropore is circular hole, the circular hole aperture is 20 μm.
The present invention also provides a kind of methods for making the disposable nanometer aperture biosensor, include the following steps：
S1, according to preset pattern, be substrate and head cover by the injection molding of medical plastic base material, and be molded into the head cover
Type processing and fabricating sample restricted area, at the same inside the sample restricted area molding making insulation cover, it is described insulation cover
Micropore is made on cover piece, the insulation cover top surface is less than sample restricted area sidewall surfaces；
S2, electrode layer is prepared in the substrate surface；
S3, the substrate surface lid for having electrode layer in preparation close head cover, and substrate and head cover are welded and fixed.
Preferably, the electrode layer is prepared by film vapor deposition technology.
Alternatively, preferably, the electrode layer is prepared by being embedded in filamentary silver to substrate.
Alternatively, preferably, the electrode layer is prepared by covering the silver foil of special pattern in substrate surface.
Preferably, the step of further including to micropore injection nano-pore and phospholipid membrane.
The above technical solution of the present invention has the following advantages over the prior art：
(1) disposable nanometer aperture biosensor of the present invention, including stacked above one another setting substrate, electrode layer and
Head cover, the head cover are provided with a diameter of 5-200 μm of micropore, and the micropore is used to inject nano-pore and phospholipid membrane.It uses
When nano-pore and phospholipid membrane are injected from micropore, obtain the biosensor of functionalization, when large biological molecule passes through nano-pore,
Due to the difference of the properties such as size and charge, specific current signature is generated, so as to according to the electric current letter acquired together
Number analyze the information such as structure of biological macromolecule, type, the concentration in sample.The nano-pore biosensor structure is simple, inspection
Survey it is convenient, of low cost, add in reagent section can it is instant plus, ensure the stability of nano-pore and phospholipid membrane itself, especially
Suitable for disposable occasion, avoid and reuse sensor and lead to interfering with each other for testing result between patient, carry
The high accuracy of testing result.
(2) production method of disposable nanometer aperture biosensor of the present invention, passes through injection molding process first
The substrate and head cover of plastic material are made, and makes hollow out position at the top of head cover and is limited with being formed for limiting the sample of detection sample
Area processed and the insulation cover for accommodating instillation reagent, making instillation reagent covering micropore and electrode layer, insulation cover are less than
Sample restricted area sidewall surfaces, it is therefore prevented that reagent overflows, by vapor deposition or other means in substrate surface according to the electrode of design
Figure prepares electrode layer, then welds substrate and head cover, forms sensor stand.The substrate and head cover used is plastics material
Matter, and by traditional injection molding process compression moulding, can high-volume quickly produce, electrode prepare before by structure design,
Reduce the area of silver coating under the premise of requirement is reached, greatly reduce the production cost of nanometer aperture biosensor, fit
Close mass industrial production and wide popularization and application.Process steps are simple, easily operated, reduce nano biological sensor
Single use cost.
Description of the drawings
In order to make the content of the present invention more clearly understood, it below according to specific embodiments of the present invention and combines
Attached drawing, the present invention is described in further detail, wherein
Fig. 1 is the vertical view of the disposable nanometer aperture biosensor of the embodiment of the present invention；
Fig. 2 is the structure diagram of the disposable nanometer aperture biosensor described in the embodiment of the present invention.
Reference numeral is expressed as in figure：1- substrates；2- electrode layers；3- head covers；31- samples restricted area；32- insulation coverings
Part；33- micropores.
The present invention can be implemented with many different forms, should not be construed as limited to embodiment set forth herein, on the contrary,
There is provided these embodiments so that the disclosure is thorough and complete, and will fully convey the concept of the invention to art technology
Personnel, the present invention will be defined by the claims.In the accompanying drawings, for clarity, the size of each device and opposite can be exaggerated
Size.
Specific embodiment
Embodiment
The present embodiment provides a kind of disposable nanometer aperture biosensors, can be used for DNA sequencing, tumor marker quality testing
Many application fields such as survey, detection of heavy metal ion and chemical analysis.
The disposable nanometer aperture biosensor is as shown in Figure 1, it includes the plastics of stacked above one another setting from the bottom to top
Substrate 1, silver electrode layer 2 and plastic top cover 3, the plastic top cover 3 has one to be used to accommodate by injection molding preparation and limitation is treated
The sample restricted area 31 of sample is detected, the sample restricted area 31 is the open hollow out position of one be made in head cover 3, hollow out
Sample is limited in inside sample restricted area 31 by four walls of position, it is therefore prevented that sample leaks, and 31 center of sample restricted area is provided with
One for accommodating the insulation cover 32 of the reagent instilled, and in this implementation, the insulation cover 32 is a cuboid plastic article,
Its described insulation cover 32 is integrally formed with the head cover, and its top surface is less than the top surface of 31 side wall of sample restricted area, prevents
Reagent in insulation cover 32 overflows.It is provided with to inject nanometer into biosensor in the insulation cover 32
Hole and the micropore 33 of phospholipid membrane, the micropore can be the shapes such as circular hole or square hole, preferably circular hole, and institute in the present embodiment
It is 5-200 μm to state the aperture of micropore 33, and preferably 20 μm in the present embodiment, the micropore 33 is through-hole, the electrode with lower section
Layer 2 connects.
Further, the silver electrode layer 2 is made of a pair of of silver electrode, and the insulation cover 32 is covered in therein one
A silver electrode surface, and the micropore 33 is arranged at the top of the silver electrode, and the silver electrode is made only to be exposed by the micropore 33,
Play the role of insulation covering.
The overall dimensions of the disposable nanometer aperture biosensor are not less than 5mm*5mm, are preferably in the present embodiment
2cm*1cm, the diameter of 31 range of sample restricted area is not less than 1.5mm, and in the present embodiment, sample restricted area 31 is rectangle,
Its single side size is not less than 2mm.
The present embodiment also provides a kind of method for making the disposable nanometer aperture biosensor, including following step
Suddenly：
S1, according to preset shape, medical plastic base material is made as substrate 1 and head cover 3, and lead to by injection molding process
It crosses injection molding process to be processed head cover 3, be limited by sample of the injection molding process in 3 internal production of head cover, one hollow out
Area 31, the sample restricted area 31, four walls are enclosed outside sample restricted area 31, form dam configuration, it is therefore prevented that and sample overflows,
Insulation cover 32 is prepared by injection molding process in the middle position of sample restricted area 31, makes 32 table of insulation cover
Face is less than 31 sidewall surfaces of sample restricted area, prevents the reagent being added dropwise from overflowing, and uses and is molded into the insulation cover 32
Type technique makes the micropore 33.
S2, silver electrode layer 2 is prepared by film vapor deposition technique on 1 surface of substrate or is used as disposable embodiment,
Can also silver electrode layer be used as by filamentary silver embedded in plastic supporting base 1 or be used as disposable embodiment, can also passed through
The silver foil of special pattern is covered as silver electrode layer in substrate surface.
S3, the head cover 3 that is prepared through step S1 of 1 surface cover of substrate conjunction that has electrode layer 2, the insulation of the head cover 3 are being prepared
Covering 32 is covered in a silver electrode surface, and micropore 33 is made to be placed at the top of the silver electrode, using thermal weld by substrate 1 with pushing up
Lid 2 is welded and fixed, and forms sensor stand, and as disposable embodiment, other welding means, which also can be used, to be served as a contrast
Bottom 1 is welded and fixed with head cover 2.It is detected in use, inject nano-pore and phospholipid membrane from the micropore 33, forms function
The nanometer aperture biosensor of change.
Disposable nanometer aperture biosensor described in the present embodiment, can high-volume quick Fabrication by injection molding process
Substrate 1 and head cover 3 by designing the structure of electrode layer 2, maximum can reduce silver-plated area, greatly under the premise of requirement is reached
The production cost for reducing biosensor thereby reduces biological detection cost, is particularly suitable for disposable occasion,
Be intended for single use it is of low cost, and avoid reuse sensor caused by interfering with each other between different patient's testing results,
Improve the detection accuracy of sensor.
Obviously, the above embodiments are merely examples for clarifying the description, and is not intended to limit the embodiments.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And the obvious variation thus extended out or
Among changing still in the protection domain of the invention.
Claims (10)
1. a kind of disposable nanometer aperture biosensor, which is characterized in that include substrate, electrode layer and the top of stacked above one another setting
Lid, the head cover are provided with a diameter of 5-200 μm of the micropore for injecting nano-pore and phospholipid membrane.
2. disposable nanometer aperture biosensor according to claim 1, which is characterized in that the head cover has a hollow out
Sample restricted area, an insulation cover is provided in the sample restricted area, the insulation cover surface is less than the sample
The surface of this restricted area side wall, the micropore are opened in the insulation cover.
3. disposable nanometer aperture biosensor according to claim 2, which is characterized in that the substrate and the head cover
It is plastic layer, the electrode is the silver electrode layer for being set to the substrate surface.
4. disposable nanometer aperture biosensor according to claim 3, which is characterized in that the silver electrode layer is by a pair
Silver electrode forms, and the insulation cover is covered in a silver electrode surface, the micropore setting in the insulation cover
At the top of silver electrode, the silver electrode is made only to be exposed by the micropore.
5. disposable nanometer aperture biosensor according to claim 4, which is characterized in that the disposable nano-pore life
The size of object sensor is not less than 5mm*5mm, and the sample restricted area range diameter is not less than 1.5mm.
6. disposable nanometer aperture biosensor according to claim 5, which is characterized in that the micropore be circular hole, institute
It is 20 μm to state circular hole aperture.
A kind of 7. method for making the disposable nanometer aperture biosensor as described in claim any one of 1-6, which is characterized in that
Include the following steps：
S1, according to preset pattern, be substrate and head cover by the injection molding of medical plastic base material, and add in the head cover injection molding
Work makes sample restricted area, at the same inside the sample restricted area molding making insulation cover, in the insulation cover
Upper making micropore, the insulation cover top surface are less than sample restricted area sidewall surfaces；
S2, electrode layer is prepared in the substrate surface；
S3, the substrate surface lid for having electrode layer in preparation close head cover, and substrate and head cover are welded and fixed.
8. the production method of disposable nanometer aperture biosensor according to claim 7, which is characterized in that the electrode
Layer is prepared by film vapor deposition technology.
9. the production method of disposable nanometer aperture biosensor according to claim 7, which is characterized in that the electrode
Prepared by filamentary silver or prepared by the silver foil by covering special pattern in substrate surface by being embedded in substrate for layer.
10. the production method of disposable nanometer aperture biosensor according to claim 8 or claim 9, which is characterized in that also wrap
The step of including to micropore injection nano-pore and phospholipid membrane.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810019657.0A CN108226249B (en) | 2018-01-09 | 2018-01-09 | Disposable nanopore biosensor and manufacturing method thereof |
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| CN201810019657.0A CN108226249B (en) | 2018-01-09 | 2018-01-09 | Disposable nanopore biosensor and manufacturing method thereof |
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| CN108226249A true CN108226249A (en) | 2018-06-29 |
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| WO2020082721A1 (en) * | 2018-10-23 | 2020-04-30 | Boe Technology Group Co., Ltd. | Biosensor apparatus, method of fabricating biosensor apparatus, biosensor chip, and method of detecting target molecule |
| CN111413383A (en) * | 2020-04-20 | 2020-07-14 | 深圳市梅丽纳米孔科技有限公司 | Method for constructing double-recognition-site nanopore |
| WO2020155242A1 (en) * | 2019-01-30 | 2020-08-06 | 深圳市梅丽纳米孔科技有限公司 | Mutant nfpab nanopore, test system, manufacturing method therefor and use thereof |
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