JP2017187443A - Polymer film for analyzer, substrate for analyzer, method of manufacturing polymer film for analyzer, and method of manufacturing substrate for analyzer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a polymer film that can be used separately from a substrate, such as an Si substrate, and reduces distortion of nanopore openings.SOLUTION: A polymer film for an analyzer comprises a polymer film (11) having a nanopore (12) formed therethrough. The nanopore (12) has openings (12a, 12b) provided at surfaces (11a, 11b) of the polymer film (11), and an inner wall (12c) formed through the polymer film (11) between the openings (12a, 12b). A ratio of a diameter of a maximum circle (13) to a diameter of a minimum circle (14) of the openings (12a, 12b) is 1.274 or less.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a polymer film for an analyzer, an analyzer, a substrate for an analyzer, a method for manufacturing a polymer film for an analyzer, and a method for manufacturing a substrate for an analyzer.

  Development of analyzers for analyzing deoxyribonucleic acid (DNA), proteins, viruses, bacteria, and the like using nanometer-sized pores called nanopores is in progress.

  For example, Patent Document 1 uses a partition in which a SiN film is stacked on a silicon (Si) substrate, and then a silicon nitride (SiN) film is etched to form a circular opening of 75 nm in diameter in the SiN film. (See, for example, paragraphs [0060] to [0073] of Patent Document 1).

  Further, for example, in Patent Document 2, an electrical insulator material exemplified by silicon, glass, quartz, polydimethylsiloxane (PDMS), polytetrafluoroethylene (PTFE), polystyrene, and polypropylene is irradiated with an electron beam or an ion beam. Thus, a nucleic acid analysis device using a solid substrate on which nanopores are formed has been disclosed (see, for example, paragraphs [0021] to [0029] of Patent Document 2).

WO2013 / 137209 JP 2013-90576 A

  However, the one-particle analysis apparatus of Patent Document 1 has a problem that the Si substrate that supports the partition wall made of the SiN film is easily broken and damaged.

  In addition, in the nucleic acid analysis device of Patent Document 2, when a nanopore is formed by irradiating a polymer material exemplified by PDMS, PTFE, polystyrene, or polypropylene with an electron beam or an ion beam, the opening of the nanopore is greatly distorted. There was a problem.

  The embodiment disclosed herein includes a polymer film, and the polymer film is provided with nanopores, and the nanopores are provided so as to penetrate through the polymer film from the openings provided on the surface of the polymer film. A polymer film for an analyzer, wherein the ratio of the diameter of the maximum circle to the diameter of the minimum circle of the opening is 1.274 or less.

  The embodiment disclosed herein includes a polymer film, and the polymer film is provided with nanopores, and the nanopores are provided so as to penetrate through the polymer film from the openings provided on the surface of the polymer film. And an inner wall, and the thickness of the polymer film is 1 μm or less.

  The embodiment disclosed herein is a substrate for an analysis apparatus including the polymer film for an analysis apparatus and a reinforcing material on the polymer film for the analysis apparatus.

  Embodiment disclosed here is an analyzer provided with said polymer film for analyzers, or said board | substrate for analyzers.

  Embodiments disclosed herein include placing a conductive material on a substrate, placing a polymer film on the conductive material, forming nanopores in the polymer film, and removing the conductive material. The nanopore has an opening provided on the surface of the polymer film and an inner wall provided so as to penetrate the polymer film from the opening. It is a manufacturing method of a polymer film.

  The embodiment disclosed herein includes a step of installing a soluble material on a substrate, a step of installing a polymer film having a thickness of 1 μm or less on the soluble material, a step of forming nanopores in the polymer film, Separating the polymer film from the substrate by dissolving, the nanopore comprising an opening provided in the surface of the polymer film and an inner wall provided to penetrate the polymer film from the opening It is the manufacturing method of the polymer film for apparatuses.

  An embodiment disclosed herein includes a step of manufacturing a polymer film for an analytical device by the above-described method for manufacturing a polymer membrane for an analytical device, and a step of installing a reinforcing material on the polymer film for the analytical device. It is a manufacturing method of the board | substrate for apparatuses.

  It is possible to provide a polymer film for an analyzer that can be used separately from a substrate such as a Si substrate and can reduce distortion of the opening of the nanopore.

2 is a schematic plan view of a polymer film for an analyzer according to Embodiment 1. FIG. It is typical sectional drawing in alignment with II-II of FIG. It is a typical enlarged plan view of the 1st opening of a nanopore, or the 2nd opening. (A) is a typical top view of an example when the conductive material installed on the board | substrate is seen from upper direction, (b) is typical sectional drawing in alignment with IVb-IVb of (a). is there. (A) is a typical top view of an example when the polymer film installed on the electroconductive material is seen from the upper part, (b) is typical sectional drawing along Vb-Vb of (a). It is. (A) is a typical top view of an example when the reinforcing material installed on the polymer film is seen from above, and (b) is a schematic cross-sectional view along VIb-VIb in (a). is there. (A) is a schematic top view of an example when the reinforcing material installed on the polymer film after formation of nanopores is viewed from above, and (b) is a schematic view along VIIb-VIIb in (a). FIG. (A) is a schematic plan view of an example when the reinforcing material on the polymer film separated from the substrate is viewed from above, and (b) is a schematic cross section taken along VIIIb-VIIIb in (a). FIG. It is a photograph of an example of the opening of the nanopore of the polymer film for analyzers of Embodiment 1 formed by FIB irradiation in the state in which the polymer film was installed on the conductive material. It is a photograph of other examples of the opening of the nanopore of the polymer film for analyzers of Embodiment 1 formed by irradiation of FIB in the state where the polymer film was installed on the conductive material. It is a photograph of an example of the opening of the nanopore formed by irradiation of FIB in the state which installed the polymer film directly on the board | substrate, without using an electroconductive material. It is a photograph of the other example of the opening of the nanopore formed by irradiation of FIB in the state which installed the polymer film directly on the board | substrate, without using an electroconductive material. It is a photograph of an example of the opening of the nanopore whose maximum / minimum circle diameter ratio is 1.026. It is a photograph of an example of the opening of the nanopore whose maximum / minimum circle diameter ratio is 1.274. It is a photograph of an example of the opening of the nanopore whose maximum / minimum circle diameter ratio is 1.105. 6 is a schematic cross-sectional view of a polymer film for an analyzer according to Embodiment 2. FIG. (A) is a typical top view of an example when the polymer film installed on the soluble material is seen from above, and (b) is a schematic cross-sectional view along XVIIb-XVIIb in (a). is there. (A) is a schematic plan view of an example when the reinforcing material installed on the polymer film is viewed from above, and (b) is a schematic cross-sectional view taken along XVIIIb-XVIIIb in (a). is there. (A) is a schematic top view of an example when the reinforcing material installed on the polymer film after formation of nanopores is viewed from above, and (b) is a schematic diagram along XIXb-XIXb in (a). FIG. It is a typical block diagram of the analyzer of Embodiment 3. It is a figure which shows an example of the change of the electric current value as a detection signal measured with the analyzer of Embodiment 3. FIG. 6 is a schematic configuration diagram of an analyzer according to a fourth embodiment. It is a figure which shows an example of the relationship between the tunnel current value obtained in the analyzer of Embodiment 4, and measurement time. FIG. 10 is a schematic cross-sectional view illustrating a part of the manufacturing process of an example of a method for manufacturing a polymer film provided with an electrode used in the analyzer of Embodiment 4. FIG. 10 is a schematic cross-sectional view illustrating another part of the manufacturing process of an example of the manufacturing method of the polymer film provided with the electrode used in the analyzer of Embodiment 4.

  Hereinafter, embodiments will be described. In the drawings used to describe the embodiments, the same reference numerals represent the same or corresponding parts.

<Embodiment 1>
In FIG. 1, the typical top view of the polymer film for analyzers of Embodiment 1 is shown. As shown in FIG. 1, the polymer film for analyzer of Embodiment 1 includes a polymer film 11 and nanopores 12 provided on the polymer film 11.

  Examples of the polymer film 11 include polymethyl methacrylate (PMMA), cycloolefin polymer (COP), cycloolefin copolymer (COC), dimethylpolysiloxane (PDMS), polycarbonate (PC), polyethylene terephthalate (PET), and polypropylene (PP ), Polytetrafluoroethylene (PTFE), polyimide (PI), polystyrene (PS), polylactic acid (PLA), SU-8, and an organic polymer film containing at least one selected from the group consisting of graphene, etc. Can be used.

  FIG. 2 shows a schematic cross-sectional view along II-II in FIG. As shown in FIG. 2, the nanopore 12 is formed on the first opening 12 a provided in the first surface 11 a of the polymer film 11 and the second surface 11 b opposite to the first surface 11 a of the polymer film 11. Provided between the first opening 12a and the second opening 12b so as to penetrate between the provided second opening 12b and the first surface 11a and the second surface 11b of the polymer film 11. And an inner wall 12c.

  In the polymer membrane for an analyzer of Embodiment 1, the ratio of the diameter of the maximum circle to the diameter of the minimum circle of at least one of the first opening 12a and the second opening 12b of the nanopore 12 (hereinafter referred to as “maximum / minimum” Is referred to as “circle diameter ratio”) is 1.274 or less.

  FIG. 3 shows a schematic enlarged plan view of the first opening 12 a or the second opening 12 b of the nanopore 12. A virtual circle having a diameter D1 of the longest length of line segments passing through the center of the first opening 12a or the second opening 12b of the nanopore 12 and connecting any two points on the outer periphery of the opening. The maximum circle is 13. In addition, the shortest length of line segments that pass through the center of the first opening 12a or the second opening 12b of the nanopore 12 and connect any two points on the outer periphery of the opening is a diameter D2. Let the circle be the minimum circle 14. The maximum / minimum circle diameter ratio is the ratio of the diameter D1 of the maximum circle 13 to the diameter D2 of the minimum circle 14 (D1 / D2).

  The longest length of a line segment (nanopore diameter) connecting any two points on the periphery of the opening through the center of the opening of the first opening 12a or the second opening 12b of the nanopore 12 is less than 1000 nm. In this case, for example, the polymer membrane for an analyzer of the first embodiment can be used for virus inspection, intermolecular interaction analysis, RNA sequencer, DNA sequencer and the like. Further, when the nanopore diameter is larger than 1000 nm, for example, the polymer membrane for analyzer of Embodiment 1 can be used for a cell sorter, a one-cell counter, and the like. In addition, when the nanopore diameter is larger than 1000 nm, two different particles having a diameter of 1000 nm or more can be separated using the polymer film for analyzer of the first embodiment. For example, when two particles having different diameters of 1000 nm or more (first particle (diameter: d1) and second particle (diameter: d2)) are separated, the nanopore diameter satisfying the relationship of d1> dp> d2 By setting dp and passing only the second particles through the nanopore 12, the first particles and the second particles can be separated.

  Hereinafter, an example of a method for producing a polymer film for an analyzer according to Embodiment 1 will be described with reference to FIGS. First, as shown in FIGS. 4A and 4B, a step of installing a conductive material 22 on the substrate 21 is performed. FIG. 4A is a schematic plan view of an example when the conductive material 22 installed on the substrate 21 is viewed from above, and FIG. 4B is a schematic view of IVb-IVb in FIG. It is typical sectional drawing along.

  The step of installing the conductive material 22 on the substrate 21 is performed, for example, by applying a solution obtained by dissolving the conductive material 22 in a solvent on a substrate such as a Si substrate, and then spin-coating and removing the solvent. it can.

  Next, as shown in FIGS. 5A and 5B, a step of installing the polymer film 11 on the conductive material 22 is performed. FIG. 5A is a schematic plan view of an example when the polymer film 11 placed on the conductive material 22 is viewed from above, and FIG. 5B is Vb-Vb in FIG. It is typical sectional drawing in alignment with.

  The step of installing the polymer film 11 on the conductive material 22 includes, for example, applying a solution obtained by dissolving a polymer, which is a raw material of the polymer film 11, in a solvent, and then spin-coating and removing the solvent. Can be done.

  In addition, the step of installing the polymer film 11 on the conductive material 22 includes the step of heating the polymer that is the raw material of the polymer film 11 to a melting point or higher to form a liquid polymer, and applying the liquid polymer on the conductive material 22. You may carry out by spin-coating and cooling a liquid polymer. Further, the step of installing the polymer film 11 on the conductive material 22 may be performed by sticking an adhesive polymer sheet (polymer film 11) on the conductive material 22.

  Next, as shown in FIGS. 6A and 6B, a step of installing a reinforcing material 24 having an opening 25 on the polymer film 11 is performed. FIG. 6A is a schematic plan view of an example when the reinforcing member 24 installed on the polymer film 11 is viewed from above, and FIG. 6B is a view taken along VIb-VIb in FIG. It is typical sectional drawing along.

  The step of installing the reinforcing member 24 having the opening 25 on the polymer film 11 is performed by, for example, irradiating the surface of the polymer film 11 on the reinforcing member 24 side by irradiating ultraviolet light after the reinforcing member 24 is installed on the polymer film 11. The activation can be performed by bonding the reinforcing material 24 to the polymer film 11. As the reinforcing material 24, for example, an acrylic plate thicker than the polymer film 11 can be used. The reinforcing material 24 is not necessarily used. Further, the reinforcing material 24 is not limited to the purpose of reinforcement. For example, when the analytical device is manufactured by attaching the polymer film for analyzer of Embodiment 1 provided with the nanopore 12 to a plate having a flow path, the reinforcing material 24 having a flow path (the plate having the flow path is reinforced). Can also be efficiently manufactured by forming nanopores 12 on the polymer film 11. That is, after the nanopore 12 is formed on the polymer film 11 to form the analytical apparatus polymer film of Embodiment 1, the process of attaching the analytical apparatus polymer film of Embodiment 1 to the plate having the flow path can be omitted.

  Next, as shown in FIGS. 7A and 7B, a step of forming nanopores 12 on the polymer film 11 is performed. FIG. 7A is a schematic plan view of an example when the reinforcing material 24 installed on the polymer film 11 after the formation of the nanopore 12 is viewed from above, and FIG. 7B is a plan view of FIG. It is typical sectional drawing in alignment with VIIb-VIIb.

  The step of forming the nanopore 12 on the polymer film 11 can be performed, for example, by irradiating the surface of the polymer film 11 exposed from the opening 25 of the reinforcing material 24 with an electron beam or an ion beam. From the viewpoint of forming the nanopore 12 with good reproducibility, it is preferable to form the nanopore 12 by irradiating a focused ion beam (FIB). In addition, the size of the opening 25 of the reinforcing material 24 may be 100 times or more the size of the opening of the nanopore 12. In addition, when the nanopore 12 is formed by irradiating the surface of the polymer film 11 with the beam, the conductive material 22 may be irradiated with the beam to form a hole in the conductive material 22. The hole may not be formed in the conductive material 22.

  Next, as shown in FIGS. 8A and 8B, a step of separating the polymer film 11 from the substrate 21 by removing the conductive material 22 is performed. FIG. 8A is a schematic plan view of an example when the reinforcing member 24 on the polymer film 11 separated from the substrate 21 is viewed from above, and FIG. 8B is a diagram of VIIIb in FIG. 8A. It is typical sectional drawing along -VIIIb.

  The step of separating the polymer film 11 from the substrate 21 by removing the conductive material 22 can be performed, for example, by dissolving the conductive material 22 with a solvent capable of dissolving the conductive material 22. For example, when the polymer film 11 and the reinforcing material 24 are not water-soluble and the conductive material 22 is water-soluble, the conductive material 22 is dissolved in water by washing with pure water, thereby changing the conductive material 22. Can be removed. By the above, the polymer film for analyzers of Embodiment 1 can be manufactured.

  Conventionally, when an organic polymer material, which is an insulating material, is irradiated with an electron beam or an ion beam to form a nanopore, the irradiation position of the beam drifts due to charging of the surface of the organic polymer material, and the nanopore The aperture was greatly distorted and the maximum / minimum circle diameter ratio was greater than 1.274. Such a distortion of the opening of the nanopore causes a variation in the opening shape of the nanopore, which in turn leads to a variation in the analysis result of the analyzer using the nanopore.

  However, in the first embodiment, since the polymer film 11 is disposed on the conductive material 22, the charge on the surface of the polymer film 11 can be discharged to the outside of the polymer film 11 through the conductive material 22. As a result, the amount of charge on the surface of the polymer film 11 when the nanopore 12 is formed by beam irradiation can be reduced as compared with the prior art. Distortion can be reduced, and as a result, variation in analysis results of the analyzer using the polymer film for analyzer of Embodiment 1 can be reduced, and analysis accuracy can be improved.

  FIG. 9 and FIG. 10 show photographs of an example of the opening of the nanopore 12 of the polymer film for analyzer of Embodiment 1 formed by FIB irradiation in a state where the polymer film 11 is placed on the conductive material 22. FIGS. 11 and 12 show photographs of examples of nanopore openings formed by FIB irradiation in a state where the polymer film 11 is directly placed on the substrate 21 without using the conductive material 22.

  As is clear from a comparison between FIG. 9 and FIG. 10 and FIG. 11 and FIG. 12, the opening of the nanopore 12 of the polymer film for analyzer according to the first embodiment is the same as that of the FIB with the polymer film directly placed on the substrate. It can be seen that the distortion can be reduced more than the opening of the nanopore formed by irradiation.

  From the viewpoint of reducing the distortion of the aperture of the nanopore 12 to reduce the variation of the aperture shape and the analysis result of the analyzer using the polymer film for analyzer of Embodiment 1, the maximum / minimum circle diameter ratio Is more preferably 1.105 or less, and further preferably 1.026 or less.

  FIG. 13 shows a photograph of an example of the opening of the nanopore 12 having a maximum / minimum circle diameter ratio of 1.026. In the example shown in FIG. 13, the diameter D1 of the maximum circle 13 is 395 nm, and the diameter D2 of the minimum circle 14 is 385 nm.

  FIG. 14 shows a photograph of an example of the opening of the nanopore 12 having a maximum / minimum circle diameter ratio of 1.274. In the example shown in FIG. 14, the diameter D1 of the maximum circle 13 is 172 nm, and the diameter D2 of the minimum circle 14 is 135 nm.

  FIG. 15 shows a photograph of an example of the opening of the nanopore 12 having a maximum / minimum circle diameter ratio of 1.105. In the example shown in FIG. 15, the diameter D1 of the maximum circle 13 is 210 nm, and the diameter D2 of the minimum circle 14 is 190 nm.

  In addition, the polymer film for analyzer of Embodiment 1 can use the polymer film 11 provided with the nanopores 12 as it is in the analyzer, so that it does not require a Si substrate as a support substrate like a SiN film. Naturally, it is less likely to be damaged than the Si substrate. In addition, you may use for the analyzer in the state of the board | substrate for analyzers which provided the reinforcing material 24 in the polymer film 11.

  Further, in the first embodiment, the nanopore 12 can be accurately formed on the polymer film 11 made of an organic polymer material such as a resin, and the cost of the analyzer can be reduced by expanding the selectivity of the material. It becomes. Moreover, compared with the case where the nanopore 12 is formed on an inorganic material such as a SiN film, the formation time of the nanopore 12 can be shortened, and the inner wall 12c of the nanopore 12 can be hydrophilized. By performing the hydrophilic treatment on the inner wall 12c of the nanopore 12, the water containing the analysis target easily passes through the inner wall 12c of the nanopore 12.

  The hydrophilic treatment of at least a part of the inner wall 12c of the nanopore 12 can be performed, for example, by irradiating at least one of the inner wall 12c of the nanopore 12 with at least one of oxygen plasma and ultraviolet light.

  Moreover, in an analyzer using nanopores, the film on which nanopores are formed may be required to be a thin film having a thickness of about several tens of nanometers. Thus, such a thin film is useful.

<Embodiment 2>
FIG. 16 is a schematic cross-sectional view of the polymer film for an analyzer according to the second embodiment. The polymer film for an analyzer according to the second embodiment is characterized in that the thickness of the polymer film 11 is 1 μm or less. As a result of intensive studies by the present inventors, when the thickness of the polymer film 11 is 1 μm or less, when the nanopore 12 is formed by the beam irradiation, before the beam irradiation position drifts, This is because it has been found that nanopores 12 having reduced opening distortion are formed after the penetration is completed.

  Hereinafter, an example of a method for producing a polymer film for an analyzer according to Embodiment 2 will be described with reference to FIGS. First, as shown in FIGS. 17A and 17B, a process of installing the soluble material 26 on the substrate 21 is performed, and a process of installing the polymer film 11 on the soluble material 26 is performed. FIG. 17A is a schematic plan view of an example when the polymer film 11 placed on the soluble material 26 is viewed from above, and FIG. 17B is an XVIIb-XVIIb in FIG. It is typical sectional drawing along.

  The step of installing the soluble material 26 on the substrate 21 can be performed, for example, by applying a solution obtained by dissolving the soluble material 26 in a solvent on a substrate such as a Si substrate, and then spin-coating and removing the solvent.

  The step of installing the polymer film 11 on the soluble material 26 is performed, for example, by applying a solution obtained by dissolving a polymer that is a raw material of the polymer film 11 in a solvent to the soluble material 26 and then spin-coating and removing the solvent. be able to.

  The step of installing the polymer film 11 on the soluble material 26 includes, for example, heating the polymer as a raw material of the polymer film 11 to a melting point or higher to form a liquid polymer, and applying the liquid polymer on the soluble material 26 You may carry out by spin-coating and cooling a liquid polymer. In addition, for example, the polymer film 11 may be placed on the soluble material 26 by sticking an adhesive polymer sheet (polymer film 11) on the soluble material 26.

Note that, unlike the first embodiment, the soluble material 26 may not be a conductive material.
Next, as shown in FIGS. 18A and 18B, a step of installing a reinforcing material 24 having an opening 25 on the polymer film 11 is performed. FIG. 18A is a schematic plan view of an example when the reinforcing member 24 installed on the polymer film 11 is viewed from above, and FIG. It is typical sectional drawing along. This step can be performed, for example, in the same manner as in the first embodiment.

  Next, as shown in FIGS. 19A and 19B, a process of forming nanopores 12 on the polymer film 11 is performed. FIG. 19A is a schematic plan view of an example when the reinforcing material 24 installed on the polymer film 11 after the nanopore 12 is formed is viewed from above, and FIG. 19B is a plan view of FIG. Is a schematic cross-sectional view along XIXb-XIXb. This step can also be performed in the same manner as in the first embodiment, for example. In addition, when forming the nanopore 12 by irradiating the surface of the polymer film 11 with the beam, the soluble material 26 may be irradiated with the beam to form holes in the soluble material 26. It is not necessary to form a hole.

  As described above, in the second embodiment, since the thickness of the polymer film 11 is 1 μm or less, the penetration of the polymer film 11 by the beam is completed and the nanopore 12 is formed before the irradiation position of the beam drifts. . Thereby, the drift of the irradiation position of the beam at the time of forming the nanopore 12 in the beam irradiation can be suppressed, and the distortion of the opening of the nanopore 12 can be reduced.

  Thereafter, a step of separating the polymer film 11 from the substrate 21 by dissolving the soluble material 26 is performed. In this step, for example, as in the first embodiment, the polymer film 11 and the reinforcing material 24 are not dissolved, but can be removed by dissolving the soluble material 26 with a solvent capable of dissolving the soluble material 26. By the above, the polymer film for analyzers of Embodiment 2 can be manufactured.

  Since the description other than the above in the second embodiment is the same as that in the first embodiment, the description thereof is omitted.

<Embodiment 3>
FIG. 20 shows a schematic configuration of the analyzer according to the third embodiment. As shown in FIG. 20, the analyzer according to the third embodiment includes an analyzer substrate 27 including a polymer film 11 provided with nanopores 12 and a reinforcing member 24 bonded to the polymer film 11. Further, the analyzer of the third embodiment includes a first chamber 34 and a second chamber 41 that are partitioned by the analyzer substrate 27.

  A first electrode 31 is provided in the first chamber 34, and the first electrode 31 is provided such that a part of the first electrode 31 is exposed inside the first chamber 34. . The first electrode 31 is connected to the ground 36 through the lead 35.

  A second electrode 32 is provided in the second chamber 41, and the second electrode 32 is provided such that a part of the second electrode 32 is exposed inside the second chamber 41. . The second electrode 32 is connected to the ground 37 through the lead 40. The ammeter 39 and the power source 38 are connected to the lead 40 in this order from the second electrode 32 side.

  The analysis by the analyzer of Embodiment 3 can be performed as follows, for example. First, the inside of the first chamber 34 and the inside of the second chamber 41 are filled with liquid, respectively. The liquid can move through the nanopore 12 between the inside of the first chamber 34 and the inside of the second chamber 41. The liquid inside the first chamber 34 contains particles 33 as an analysis target. The first electrode 31 is in contact with the liquid filled in the first chamber 34, and the second electrode 32 is in contact with the liquid filled in the second chamber 41.

  The liquid that fills the inside of the first chamber 34 and the inside of the second chamber 41 may be a liquid that can energize between the first electrode 31 and the second electrode 32. As the liquid that can be energized, for example, an electrolyte solution such as a KCl aqueous solution, or a buffer solution such as a TE buffer solution or a PBS buffer solution can be used.

  A current is passed between the first electrode 31 and the second electrode 32 by the power supply 38 in a state where the liquid is filled in the first chamber 34 and the second chamber 41. A current value between the first electrode 31 and the second electrode 32 is measured with an ammeter 39 over time. The particles 33 in the liquid inside the first chamber 34 move by diffusion to the liquid inside the second chamber 41 through the nanopore 12. The current value measured by the ammeter 39 when the particle 33 passes through the nanopore 12 decreases depending on the size and shape of the particle 33.

  FIG. 21 shows an example of a change in current value as a detection signal measured by the analyzer of the third embodiment. The vertical axis in FIG. 21 shows the current value, and the horizontal axis in FIG. 21 shows the measurement time. In a state where the particles 33 are not present in the nanopore 12, the current value between the first electrode 31 and the second electrode 32 shows a constant current value 51. This current value 51 is a base value. When the particle 33 passes through the nanopore 12, the current value between the first electrode 31 and the second electrode 32 becomes a current value 52 that decreases depending on the size and shape of the particle 33. After the particle 33 has completely passed through the nanopore 12, the current value 53 is restored to the level before the particle 33 passes through the nanopore 12.

  This phenomenon can be explained as follows. When the particles 33 pass through the nanopore 12, the current-carrying substance in the liquid that can be energized, such as ions in the nanopore 12 that is the current path, is excluded by the amount of the particles 33. Thereby, the current resistance increases, and as a result, the current value decreases. When the passage of the particles 33 is completed, the current value is restored to the original level. If one particle 33 passes, one current drop signal is observed. If the current drop signal is counted, the number of particles that have passed can be counted. Here, the amount of ions excluded in the nanopore 12 due to the presence of the particles 33 depends on the size and shape of the particles 33.

<Embodiment 4>
FIG. 22 shows a schematic configuration of the analyzer according to the fourth embodiment. The analyzer according to the fourth embodiment includes a container 65. The inside of the container 65 is composed of a first solution tank 66 and a second solution tank 67 by a first electrode 61 and a second electrode 62 and two analyzer substrates 27 sandwiching these electrodes. It is divided into.

  The first solution tank 66 includes an introduction port 68 on one side surface and a discharge port 69 on the other side surface. The first solution tank 66 includes first nucleic acid driving means 63 between the inlet 68 and the outlet 69.

  The second solution tank 67 includes an introduction port 70 on one side surface and a discharge port 71 on the other side surface. The second solution tank 67 includes a second nucleic acid driving means 64 between the introduction port 70 and the discharge port 71.

  A power source (not shown) and an ammeter (not shown) are connected between the first electrode 61 and the second electrode 62. The power supply, ammeter, first nucleic acid driving means 63, and second nucleic acid driving means 64 are connected to control and data processing means (not shown).

  The analysis by the analyzer of Embodiment 4 can be performed as follows, for example. First, when analyzing the nucleic acid 72, the nucleic acid 72 to be analyzed is mixed with the buffer solution and introduced into the first solution tank 66 through the inlet 68. Further, only the buffer solution is introduced into the second solution tank 67 through the introduction port 70.

  Next, the nucleic acid 72 is moved from the first solution tank 66 to the second solution tank 67 by driving the first nucleic acid driving means 63 and the second nucleic acid driving means 64. Simultaneously with the movement of the nucleic acid 72, a voltage is applied between the first electrode 61 and the second electrode 62 by the power source, and the current flowing between the first electrode 61 and the second electrode 62 is measured by an ammeter. Use to measure. When the nucleic acid 72 is introduced into the nanopore 12, the current value decreases. When the current value decreases, the first nucleic acid driving unit 63 and the second nucleic acid driving unit 64 are configured so that the base species can be identified by the current flowing between the first electrode 61 and the second electrode 62. Thus, the speed at which the nucleic acid 72 passes through the nanopore 12 is adjusted. The base type is identified by a difference in current value flowing under a constant voltage or a difference in current-voltage characteristics of each base.

  For example, an example of a method for identifying a base of a DNA strand based on a difference in current value flowing under a constant voltage using the analyzer according to the fourth embodiment will be described. First, a tunnel current value when a polymer in which only one type of DNA chain base ATGC is continuous passes through the nanopore 12 is measured in advance. There are four types of bases in the DNA chain, ATGC. A unique current value is measured for each type of these bases and stored in the memory of the control and data processing means.

  Next, the tunnel current when the DNA strand to be analyzed passes through the nanopore 12 is actually measured, and the measured tunnel current value and the current corresponding to the type of each base of ATGC stored in the memory in advance. Compare the value. This makes it possible to identify the bases constituting the DNA strand to be analyzed.

  FIG. 23 shows an example of the relationship between the current value obtained in the analyzer of Embodiment 4 and the measurement time. The vertical axis in FIG. 23 indicates the current value, and the horizontal axis indicates the measurement time. In the example shown in FIG. 23, base A has the highest current value, base G has the second highest current value, base C has the third highest current value, and base T has the lowest current value. . And the base which comprises a DNA strand is identified by the difference in the electric current value of these bases.

  Hereinafter, an example of a method for manufacturing the polymer film 11 including the electrodes 61 and 62 used in the analyzer of Embodiment 4 will be described with reference to the schematic cross-sectional views of FIGS. 24 and 25. First, as shown in FIG. 24, the conductive material 22, the polymer film 11, the metal film 81, and the polymer film 11 are placed on the substrate 21 in this order. Here, the soluble material 26 may be installed instead of the conductive material 22 or together with the conductive material 22. The metal film 81 can be formed by depositing on the polymer film 11 by, for example, a sputtering method or a vapor deposition method and then patterning by a lithography technique or the like.

  Next, as shown in FIG. 25, for example, the polymer film 11 and the metal film 81 and the polymer film 11 are irradiated with a beam 82 such as an electron beam, an ion beam, or an FIB to thereby form the polymer film 11 and the polymer film 11. Nanopores 12 are formed on the metal film 81. By forming the nanopore 12 on the metal film 81, the electrodes 61 and 62 can be formed. Further, instead of using the conductive material 22 or using the conductive material 22 and irradiating the beam 82 with the leads 61 and 62 attached to the electrodes 61 and 62, the polymer at the time of forming the nanopore 12 is obtained. The amount of charge on the surface of the film 11 can also be suppressed.

  Thereafter, the conductive material 22 is removed, and the laminate of the polymer film 11, the electrodes 61 and 62, and the polymer film 11 is separated from the substrate 21, thereby providing the electrodes 61 and 62 used in the analyzer of Embodiment 4. A polymer film 11 can be formed. Thereafter, the reinforcing material 24 can be attached to the polymer film 11, but the reinforcing material 24 may not be attached to the polymer film 11.

  Although the embodiment has been described as described above, it is also planned from the beginning to appropriately combine the configurations of the above-described embodiments.

  It should be thought that embodiment disclosed this time is an illustration and restrictive at no points. The scope of the present invention is defined by the terms of the claims, rather than the description above, and is intended to include any modifications within the scope and meaning equivalent to the terms of the claims.

  11 polymer film, 11a first surface, 11b second surface, 12 nanopore, 12a first opening, 12b second opening, 12c inner wall, 13 maximum circle, 14 minimum circle, 21 substrate, 22 conductive material, 24 Reinforcing Material, 25 Opening, 26 Soluble Material, 27 Analyzer Substrate, 31 First Electrode, 32 Second Electrode, 33 Particles, 34 First Chamber, 35, 40 Lead, 36, 37 Ground, 38 , 39 power supply, 51, 52, 53 current value, 61 first electrode, 62 second electrode, 63 first nucleic acid driving means, 64 second nucleic acid driving means, 65 container, 66 first solution tank, 67 Second solution tank, 68, 70 inlet, 69, 71 outlet, 72 nucleic acid, 81 metal film, 82 beam.

Claims (18)

With a polymer membrane,
The polymer film is provided with nanopores,
The nanopore comprises an opening provided on the surface of the polymer film, and an inner wall provided so as to penetrate the polymer film from the opening,
The polymer membrane for an analyzer, wherein the ratio of the diameter of the maximum circle to the diameter of the minimum circle of the opening is 1.274 or less.   The polymer film for an analyzer according to claim 1, wherein the ratio is 1.105 or less.   The polymer film for an analyzer according to claim 2, wherein the ratio is 1.026 or less. With a polymer membrane,
The polymer film is provided with nanopores,
The nanopore comprises an opening provided on the surface of the polymer film, and an inner wall provided so as to penetrate the polymer film from the opening,
A polymer film for an analyzer, wherein the polymer film has a thickness of 1 μm or less.   The polymer film for an analyzer according to any one of claims 1 to 4, wherein at least a part of the inner wall is hydrophilic.   The polymer film for an analyzer according to any one of claims 1 to 5, wherein the polymer film contains polymethyl methacrylate.   A substrate for an analyzer comprising the polymer film for an analyzer according to any one of claims 1 to 6 and a reinforcing material on the polymer film for the analyzer.   The analyzer provided with the polymer film for analyzers of any one of Claims 1-6, or the board | substrate for analyzers of Claim 7. Installing a conductive material on the substrate;
Installing a polymer film on the conductive material;
Forming nanopores in the polymer film;
Separating the polymer film from the substrate by removing the conductive material;
The said nanopore is the manufacturing method of the polymer film for analyzers provided with the opening provided in the surface of the said polymer film, and the inner wall provided so that the said polymer film might be penetrated from the said opening. The conductive material is water soluble,
The method for producing a polymer film for an analyzer according to claim 9, wherein the conductive material is removed by dissolving in water.   The method for producing a polymer film for an analyzer according to claim 9 or 10, wherein a ratio of a diameter of a maximum circle to a diameter of a minimum circle of the opening is 1.274 or less.   The method for producing a polymer film for an analyzer according to claim 11, wherein the ratio is 1.105 or less.   The method for producing a polymer film for an analyzer according to claim 12, wherein the ratio is 1.026 or less. Installing a soluble material on the substrate;
Installing a polymer film having a thickness of 1 μm or less on the soluble material;
Forming nanopores in the polymer film;
Separating the polymer film from the substrate by dissolving the soluble material,
The said nanopore is the manufacturing method of the polymer film for analyzers provided with the opening provided in the surface of the said polymer film, and the inner wall provided so that the said polymer film might be penetrated from the said opening.   The method for producing a polymer film for an analyzer according to any one of claims 9 to 14, further comprising a step of hydrophilizing at least a part of the inner wall.   The method for producing a polymer film for an analyzer according to claim 15, wherein the step of hydrophilizing at least a part of the inner wall is performed by at least one of oxygen plasma treatment and ultraviolet irradiation.   The said nanopore is a manufacturing method of the polymer film for analyzers of any one of Claims 9-16 formed by irradiating a focused ion beam to the said polymer film. A process for producing a polymer film for an analyzer by the method for producing a polymer film for an analyzer according to any one of claims 9 to 17,
And a step of installing a reinforcing material on the polymer film for an analyzer.