阅读说明：本技术 一种固态纳米孔修饰处理方法 (solid-state nanopore modification treatment method ) 是由 古家强 伊戈尔·伊万诺夫 何筠 田晖 于 2019-10-11 设计创作，主要内容包括：本发明公开了一种固态纳米孔修饰处理方法,包括以下步骤：1)对固态纳米孔进行清洗和干燥；2)将清洗和干燥处理过的固态纳米孔置于等离子清洗机中,进行表面等离子处理,得到羟基化的固态纳米孔；3)将硅烷溶液涂覆在羟基化的固态纳米孔上,保护气氛下进行硅烷化,再进行清洗和干燥,得到硅烷化的固态纳米孔；4)对硅烷化的固态纳米孔进行烘烤处理、清洗和干燥。本发明在短时间内便可以对大量固态纳米孔进行快速有效的修饰处理,对实验设备条件要求简单,成本低廉。(The invention discloses a solid-state nanopore modification treatment method which comprises the following steps of 1) cleaning and drying solid-state nanopores, 2) placing the cleaned and dried solid-state nanopores in a plasma cleaning machine for surface plasma treatment to obtain hydroxylated solid-state nanopores, 3) coating a silane solution on the hydroxylated solid-state nanopores, silanizing in a protective atmosphere, cleaning and drying to obtain silanized solid-state nanopores, and 4) baking, cleaning and drying the silanized solid-state nanopores.)

1, solid-state nanopore modification treatment methods, characterized by comprising the following steps:

1) cleaning and drying the solid-state nano-pores;

2) placing the cleaned and dried solid-state nanopore in a plasma cleaning machine, and carrying out surface plasma treatment to obtain a hydroxylated solid-state nanopore;

3) coating a silane solution on the hydroxylated solid-state nano-pores, silanizing under a protective atmosphere, and then cleaning and drying to obtain silanized solid-state nano-pores;

4) and (3) baking, cleaning and drying the silanized solid-state nano pores.

2. The solid state nanopore modification treatment method of claim 1, wherein: and the cleaning in the step 1) is to sequentially clean the solid-state nano-pores by using deionized water and isopropanol.

3. The solid state nanopore modification treatment method of claim 1, wherein: and 2) the surface plasma treatment time is 2-5 min.

4. The solid state nanopore modification treatment method of claim 1, wherein: and 3) preparing the silane solution from silane and a solvent according to a volume ratio of 1: (9-99).

5. The method for modifying and treating solid-state nanopores according to any of claims 1-4, wherein the silane in the silane solution in step 3) is at least selected from 3-aminopropyldimethylethoxysilane, dimethyloctylchlorosilane, 3-mercaptopropyltriethoxysilane, 3-aminopropyltriethoxysilane, 3- (2, 3-glycidoxy) propyltrimethoxysilane, (3-cyanopropyl) dimethylchlorosilane, and dimethyloctadecylchlorosilane.

6. The method for modifying and processing solid-state nanopores according to any of claims 1-4, wherein the silane solution of step 3) is coated on the solid-state nanopores by spin coating.

7. The method for modifying and treating solid-state nanopores according to any of claims 1-4, wherein the silylation time in step 3) is 20-40 min.

8. The method for modifying and treating solid-state nanopores according to any of claims 1-4, wherein the baking treatment in step 4) is carried out at 80-100 ℃ for 20-40 min.

9. The method for modifying and treating solid-state nanopores according to any of claims 1-4, wherein the drying in steps 1), 3) and 4) is blowing with nitrogen.

10. The method for modifying and treating solid-state nanopores according to any of claims 1-4, wherein the washing in steps 3) and 4) is carried out by washing the solid-state nanopores with isopropanol.

Technical Field

The invention relates to solid-state nanopore modification treatment methods.

Background

The solid-state nanometer pores are nanometer-sized channels artificially prepared on a nanometer-scale thin film, have high stability, adjustable pore diameter and chemical environment, can be combined with a semiconductor integrated manufacturing method for large-scale production, and have huge application prospects in the aspect of single-molecule detection. Similar to coulter's principle of a cell counter, when a molecule passes through a nanopore with a similar size, the molecule can have a significant influence on ion flow in a nanopore channel, an electrochemical signal capable of being measured and analyzed appears, and the electrochemical signal is analyzed to obtain related information of a detected object.

Compared with the traditional sequencing method by using a large optical instrument, the nanopore sequencing (namely the real-time analysis of DNA single-molecule base sequences through solid-state nanopores) has the advantages of miniaturization, low cost, high sensitivity and the like, and is a hotspot of current research. The principle of nanopore sequencing is that a single DNA molecule is driven to pass through a solid nanopore with a diameter similar to that of the single DNA molecule by an electric field in an electrolyte solution, different bases (A, T, C, G) have different molecular structures, so that the influence on ion flow in the solid nanopore is different, and corresponding bases are identified by measuring corresponding base characteristic current signals during translocation of the DNA molecule, so that the base sequence reading of the DNA is realized. The solid-state nanopore can be used for gene sequencing and has great application value in other single molecule detection fields, such as: the protein detection can be used as a marker for detecting serious diseases, and the early diagnosis of the serious diseases is realized.

When biomolecules pass through the solid-state nanopore under the drive of an electric field, the translocation speed of through holes of the biomolecules is high, and the current signal acquisition instrument at the present stage cannot achieve such high time resolution (for example, the translocation speed of DNA molecules is about 5-30 nanoseconds per base, and the sampling time required by the instrument is in the level of milliseconds per base), so that the solid-state nanopore which is not subjected to any functional modification can only distinguish single chains and double chains of the DNA molecules and the difference between the long chains and the short chains.

Therefore, it is necessary to develop methods for modifying solid-state nanopores with high efficiency and high modification efficiency.

Disclosure of Invention

The invention aims to provide solid-state nanopore modification treatment methods.

The technical scheme adopted by the invention is as follows:

A solid-state nanopore modification treatment method, comprising the following steps:

1) cleaning and drying the solid-state nano-pores;

2) placing the cleaned and dried solid-state nanopore in a plasma cleaning machine, and carrying out surface plasma treatment to obtain a hydroxylated solid-state nanopore;

3) coating a silane solution on the hydroxylated solid-state nano-pores, silanizing under a protective atmosphere, and then cleaning and drying to obtain silanized solid-state nano-pores;

4) and (3) baking, cleaning and drying the silanized solid-state nano pores.

Preferably, the cleaning in step 1) is sequentially performed on the solid-state nanopore by using deionized water and isopropanol.

Preferably, the time of the surface plasma treatment in the step 2) is 2-5 min.

Preferably, the silane solution in the step 3) is prepared by mixing silane and a solvent according to a volume ratio of 1: (9-99).

Preferably, the silane in the silane solution in the step 3) is at least of 3-aminopropyldimethylethoxysilane, dimethyloctylchlorosilane, 3-mercaptopropyltriethoxysilane, 3-aminopropyltriethoxysilane, 3- (2, 3-glycidoxy) propyltrimethoxysilane, (3-cyanopropyl) dimethylchlorosilane and dimethyloctadecylchlorosilane.

Preferably, the solvent in the silane solution in step 3) is at least of decane, ethanol, acetone, hexane, octane and hexadecane.

Preferably, the silane solution in step 3) is coated on the solid-state nanopore by means of spin coating.

Preferably, the silanization time in the step 3) is 20-40 min.

Preferably, the protective atmosphere in step 3) is a nitrogen atmosphere.

Preferably, the temperature of the baking treatment in the step 4) is 80-100 ℃, and the time is 20-40 min.

Preferably, the drying in steps 1), 3) and 4) is blow-drying with nitrogen.

Preferably, the washing in steps 3) and 4) is carried out by washing the solid-state nanopore with isopropanol.

The invention has the beneficial effects that: the invention can quickly and effectively modify a large amount of solid-state nano-pores in a short time, has simple requirements on experimental equipment conditions and low cost.

Drawings

FIG. 1 is an I-V curve of solid state nanopores treated in examples 1-3 under the same conditions of potassium chloride concentration (100mM) and pH (pH 7).

FIG. 2 is a graph of conductance-pH relationship between aminosilane-modified silica solid-state nanopores and unmodified silica solid-state nanopores in a KCl solution with a concentration of 100mM in example 1.

Fig. 3 is an I-V curve of the chlorosilane-modified silicon nitride solid-state nanopore of example 3, the chlorosilane-modified silicon nitride solid-state nanopore of comparative example, and the unmodified silicon nitride solid-state nanopore under the same potassium chloride concentration (100mM) and pH (pH 7).

Detailed Description

The invention is further illustrated and described at with reference to specific examples.