WO2008016032A1

WO2008016032A1 - Filter for biochip, method for manufacturing filter for biochip and biochip using filter for biochip

Info

Publication number: WO2008016032A1
Application number: PCT/JP2007/064943
Authority: WO
Inventors: Katsuyuki Ono; Muneo Harada; Katsuya Okumura; Makoto Mihara; Mutsuhiko Yoshioka; Katsuhiko Hieda
Original assignee: Jsr Corporation
Priority date: 2006-07-31
Filing date: 2007-07-31
Publication date: 2008-02-07
Also published as: JP2008032656A

Abstract

[PROBLEMS] Provided is a filter for biochips, which has improved filter strength and durability, improved yield without filter breakage and damage in a manufacturing process, and reduced cost. A method for manufacturing the filter for biochips and a biochip using the filter for biochips are also provided. [MEANS FOR SOLVING PROBLEMS] A filter for biochips is provided with a filter main body, which is composed of a metal substrate configuring a substrate section and is made of silicon or the like; a well, which forms a recessed section on the filter main body; and a filter which configures the bottom surface of the well and has a filter having a plurality of penetrating pores. The filter is formed of a metal such as silicon.

Description

Specification

Biochip filter, biochip filter manufacturing method, and biochip using biochip filter

Technical field

[0001] The present invention relates to a target substance comprising, for example, a step of reacting or interacting with a particle carrying a bioprobe such as nucleic acid or protein, and a target substance in a specimen, and a step of washing the particle. Biochip filters and biochips used for biochips to perform B / F separation, detection and identification of target substances and bioprobes in the specimen by reaction or interaction using optical means, etc. The present invention relates to a method for producing a filter for a biochip and a biochip using a filter for a biochip.

Background art

[0002] As a method for accurately detecting a nucleic acid having a target base sequence, a nucleic acid having a sequence complementary to this nucleic acid is supported on a particle as a probe, and in a solution containing the probe-supported particle, A method of reacting probe-carrying particles with a target substance in a specimen is known (Patent Documents !! to 5).

[0003] This method has the advantage that the reactivity is very high because the probe-carrying particles are three-dimensionally dispersed in the solution and the probe-carrying particles and the analyte can move to each other. is there. For example, the target substance in the sample reacts specifically with the probe on the latex particle surface, and the content of the target substance that specifically reacts and binds to the probe is analyzed by B / F (Bound form / Free form) separation. Detection and identification methods are known.

[0004] Biochips have DNA chips or oligonucleotides immobilized thereon as probes, proteins such as antigens and antibodies, or proteins that specifically react with or interact with these proteins. There is also a chip, etc. For various applications, there is a need for a biochip capable of detecting and identifying with high sensitivity and higher B / F separation efficiency.

[0005] As such a biochip, the present applicants use a technique for using a biochip filter that allows a liquid such as an analyte and a cleaning liquid to pass therethrough and does not allow probe-carrying particles to pass therethrough. Is disclosed in Patent Document 6 (Japanese Patent Laid-Open No. 2005-148048).

[0006] That is, the biochip filter disclosed in Patent Document 6 is a plate made of a plurality of materials having different compositions, such as aluminum / alumina, metal silicon / silica, as a method for producing a biochip filter. Alternatively, a method for forming filters and wells by performing both-side forces and pattern etching on metal titanium / tita, etc. is disclosed!

[0007] Specifically, for example, a filter is formed by etching the metal oxide layer such as alumina, silica, and titania to the boundary between the metal oxide layer and the metal layer, and then aluminum, metal It is disclosed that a biochip filter in which a well and a filter are joined can be manufactured by etching a metal layer such as silicon or titanium metal to the boundary between the metal layer and the metal oxide layer.

[0008] The manufacturing method of the biochip filter disclosed in Patent Document 6 is roughly as follows.

That is, first, as shown in FIG. 8A, on both surfaces of the silicon substrate body 101, for example,

A silicon substrate 100 having a thickness of about 300 m on which a thermal oxide film 102, 104 of SiO having a thickness of about 2 ^ m is formed is prepared.

[0010] Then, as shown in FIG. 8B, a resist film 106 is formed on the upper surface of the SiO thermal oxide film 102 of the silicon substrate 100, and a fine filter is formed by photolithography. A recess 108 pattern corresponding to a hole is formed.

Next, as shown in FIG. 8C, etching removal of the SiO thermal oxide film 102 is performed through the recess 108, so that the portion of the SiO thermal oxide film 102 is removed. A recess 110 corresponding to the pore is formed. Then, as shown in FIG. 8D, the resist film 106 is peeled off.

Then, with the top and bottom turned upside down, as shown in FIG. 9A, a resist film 112 is formed on the upper surface of the thermal oxidation film 104 of S in the silicon substrate body 101, and photolithography is performed. According to one, the concave 114 pattern corresponding to the well is formed.

Next, as shown in FIG. 9B, the SiO thermal oxide film 104 is etched and removed through the recess 114, so that the portion of the SiO thermal oxide film 104 is exposed to the well. Recess 116 Form. Then, as shown in FIG. 9C, the resist film 112 is peeled off.

Then, as shown in FIG. 9C, the exposed portion of the silicon substrate body 101 is removed by etching using the portion of the thermal oxide film 104 of SiO as a mask to form the well 124.

Next, as shown in FIG. 10 (A), a dicing tape 128 is attached to the thermal oxide film 104 of SiO, and then dicing is performed as shown in FIG. 10 (B). Separate into individual biochip filters 130.

[0016] Then, as shown in FIG. 11 (A), the dicing tape 128 is expanded by pulling outward, and the dicing tape is heated to increase the temperature of the dicing tape.

Each chip is peeled from 128 and separated into individual biochip filters 130.

Accordingly, as shown in FIG. 11 (B), a filter main body 130 composed of the silicon substrate main body 101 constituting the substrate portion and a plurality of recesses provided in the filter main body 130 are formed. A biochip filter 130 is obtained which is provided with the wel 124 and the filter 136 which forms the bottom surface of the wel 124 and has a plurality of through-holes 110 formed therein.

In FIGS. 8 to 11, for the sake of convenience of explanation, the force indicating one well 124 is actually formed with a plurality of wells 124 spaced apart from each other by a fixed interval.

Patent Document 1: U.S. Pat.No. 5,736,330

Patent Document 2: Japanese Patent Laid-Open No. 62-81566

Patent Document 3: Japanese Patent Publication No. 7-54324

Patent Document 4: Japanese Patent Laid-Open No. 11 243997

Patent Document 5: Japanese Unexamined Patent Publication No. 2000-346842

Patent Document 6: Japanese Unexamined Patent Publication No. 2005-148048

Disclosure of the invention

Problems to be solved by the invention

However, in such a conventional biochip filter 130 disclosed in Patent Document 6, the filter 136 is a metal oxide that is a glass component such as alumina film, silica (silicon oxide film), titania, and the like. Although it is composed of material layers and has some strength and durability, the thickness of the filter 136 is, for example, about 1 to 10 mm, and extremely thin. For this reason, there is a need for a biochip filter with improved strength and durability that can prevent damage to the filter even when the pressure of the liquid when flowing the cleaning liquid is increased.

[0020] In view of such a current situation, the present invention further improves the strength and durability of the filter as compared with the conventional biochip filter, and the yield is improved in the manufacturing process without damage to the filter. An object of the present invention is to provide a biochip filter capable of reducing costs, a method for producing a biochip filter, and a biochip using the biochip filter.

Means for solving the problem

[0021] The present invention has been invented in order to achieve the above-described problems and objects in the prior art, and the method for producing a biochip filter of the present invention includes:

A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

A filter is provided which forms the bottom surface of the well and has a plurality of through-holes formed therein.

A method of manufacturing a filter for a biochip made of metal, S, which is the best of the filter,

Using a wafer comprising a metal substrate, a metal oxide layer formed on the upper surface of the metal substrate, and a metal thin film layer formed on the upper surface of the metal oxide layer,

The metal thin film layer and a part of the metal substrate are etched and removed from the upper and lower surfaces through the metal oxide layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. And a filter formed from.

With this configuration, the metal thin film layer and the metal substrate are etched and removed from the upper and lower surfaces through the metal oxide layer, thereby forming the well and the bottom surface of the well, and a plurality of through holes are formed. Fine pores are formed, and a filter made of metal can be formed.

[0023] Therefore, a biochip filter including a filter formed of metal can be easily manufactured without a complicated process, and the yield of the filter is not damaged in the manufacturing process. Can be improved and the cost can be reduced. [0024] In addition, the method for producing a biochip filter of the present invention includes:

Using a wafer comprising the metal substrate, a metal oxide layer formed on the upper surface of the metal substrate, and a metal thin film layer formed on the upper surface of the metal oxide layer,

A recess forming step of forming a recess in which the fine holes of the finoletor are to be formed by etching away a part of the metal thin film layer of the wafer;

Forming a well so as to face the recess through the metal oxide layer by etching away the metal substrate of the wafer;

A filter forming step of forming a filter in which a plurality of through pores are formed by communicating a well with a recess to form the pores of the filter by etching away a part of the metal oxide layer. When,

The provision of

With this configuration, in the recess forming process, the recess is formed by removing the metal thin film layer of the wafer to form a recess in which the pores of the filter are to be formed. By removing the substrate by etching, a tool is formed through the metal oxide layer so as to face the recess. Then, in the filter formation process, the metal oxide layer is removed by etching to form filter pores. It is the force to form a filter in which a plurality of fine pores are formed by communicating the recess to be welded with the well.

[0026] Therefore, a biochip filter having a filter formed of metal can be easily obtained without performing a complicated process by performing the recess forming process, the well forming process, and the filter forming process. In addition, in the manufacturing process, there is no breakage and damage of the filter, yield can be improved, and cost can be reduced.

[0027] By configuring in this way, the strength of the filter is made of, for example, a metal such as silicon. For example, the strength and durability of a very thin filter having a thickness of about 1 to 10 m are obtained. For example, this is a significant improvement over a biochip filter having a conventional filter composed of a metal oxide layer such as alumina, silica (silicon oxide film), and titania.

[0028] Therefore, the biochip filter of the present invention can be used, for example, for nucleic acid, protein, and other buffers. B / F separation of the target substance, including the step of reacting or interacting the particle carrying the bioprobe with the target substance in the specimen and the step of washing the particle, and the target substance and bioprobe in the specimen When applied to a biochip filter used in biochips for detection and identification by reaction or interaction with optical means, etc., these B / F separation and target substances in the specimen It is possible to reliably perform detection and identification by optical means or the like of the reaction or interaction between the probe and the bioprobe.

[0029] Also, when manufacturing a biochip filter, since the strength of the filter is high, the yield can be improved without damaging and damaging the filter for one minute, and the cost can be reduced.

[0030] Further, the method for producing a biochip filter of the present invention includes:

A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

The filter is formed of metal;

The metal substrate force constituting the filter body is composed of a silicon substrate,

A method for producing a biochip filter, which is a metallic silicon that constitutes the filter,

Using an SOI (Silicon On Insulator) wafer consisting of a silicon substrate, an SiO layer formed on the upper surface of the silicon substrate, and a silicon thin film layer formed on the upper surface of the SiO layer,

The silicon thin film layer and a part of the silicon substrate are etched and removed from the upper and lower surfaces through the SiO layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. Forming a filtered filter.

[0031] With this configuration, the silicon thin film layer and the silicon substrate are etched and removed from the upper and lower surfaces through the SiO layer, thereby forming the well and the bottom surface of the well, and a plurality of penetrating fine cells. A hole is formed, and a filter made of silicon can be formed. [0032] Therefore, a biochip filter equipped with a filter formed from silicon can be easily manufactured without a complicated process, and in the manufacturing process, there is no damage to the filter and the yield is high. It is possible to improve and reduce costs.

[0033] In addition, the method for producing a biochip filter of the present invention includes:

Using an SOI (Silicon On Insulator) wafer comprising the silicon substrate, a SiO layer formed on the upper surface of the silicon substrate, and a silicon thin film layer formed on the upper surface of the SiO layer, one of the silicon thin film layers of the SOI wafer is used. A recess forming step for forming a recess in which the pores of the filter are to be formed by etching away the portion;

Forming a well so as to face the recess through the SiO layer by etching away the silicon substrate of the SOI wafer;

A filter forming step of forming a filter in which a plurality of through-holes are formed by communicating a well with a recess to form a pore of the filter by etching away a part of the SiO layer; and

The provision of

With this configuration, in the recess forming process, the silicon thin film layer of the SOI wafer is etched away to form a recess in which the pores of the filter are to be formed. By etching away the silicon substrate of the wafer, a well is formed so as to face the recess through the SiO layer. Then, in the filter forming process, the SiO layer is removed by etching, and the pores of the filter are formed. This is the force by forming a filter in which a plurality of through-holes are formed by communicating the recess to be formed with the well.

[0035] Therefore, a biochip filter including a filter formed of silicon can be easily obtained without performing a complicated process by performing the recess forming process, the well forming process, and the filter forming process. In addition, in the manufacturing process, there is no breakage and damage of the filter, the yield is improved, and the chip manufacturing cost can be reduced.

[0036] Further, the method for producing a biochip filter of the present invention includes:

The SiO2 layer is etched away in the well pattern portion. [0037] With this configuration, a well corresponding to the well pattern can be formed accurately and efficiently.

[0038] Further, the method for producing a biochip filter of the present invention is characterized in that the surface of the silicon thin film layer in the tool portion is oxidized.

[0039] When the surface force of the silicon thin film layer in the well portion is oxidized in this way, particles attached to the surface can be easily removed with a solution such as diluted hydrofluoric acid, and the yield of the product is improved.

[0040] Further, the biochip filter manufacturing method of the present invention is characterized in that the ratio of the depth of the filter to the pore diameter (hereinafter referred to as "aspect ratio") is 5 or less.

[0041] When the aspect ratio of the depth to the pore diameter of the filter is 5 or less as described above, pores having a uniform pore diameter are formed over the entire surface of the wafer when etching is performed in the formation of the pores. If the production effect of being able to flow, the flow of liquid when draining the liquid through the pores is easy to flow to the extent that it does not become a practical problem is obtained.

[0042] The biochip filter manufacturing method of the present invention is characterized in that a distance force between the tools is 0.05 mm or more.

[0043] As described above, when the distance force between the wells is 0.05 mm or more, the strength of the chip can be sufficiently maintained in the metal region between the wells.

[0044] Further, the filter for a biochip of the present invention includes

A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

A biochip filter provided with a filter that forms the bottom surface of the well and has a plurality of through-holes formed therein.

The filter force S is formed of metal.

[0045] Further, the biochip filter of the present invention comprises:

The metal substrate force constituting the filter body is composed of a silicon substrate. The filter is made of metallic silicon that constitutes the filter.

[0046] With this configuration, the filter is made of a metal such as silicon. For example, the strength and durability of a very thin filter having a thickness of about 1 to 10 m are obtained. For example, this is a significant improvement over a biochip filter having a conventional filter composed of a metal oxide layer such as alumina, silica (silicon oxide film), and titania.

[0047] Therefore, the biochip filter of the present invention comprises, for example, a step of reacting or interacting with a particle carrying a bioprobe such as a nucleic acid or protein and a target substance in a specimen, and washing the particle. Biochips used in biochips to perform B / F separation of target substances including processes, and detection / identification by means of optical means of reaction or interaction between target substances and bioprobes in the specimen When applied to filters, it is possible to reliably detect and identify these B / F separations and optical means for the reaction or interaction between the target substance in the sample and the bioprobe.

[0048] Also, when manufacturing a biochip filter, since the strength of the filter is high, the yield can be improved and the cost can be reduced without damaging and damaging a part of the filter.

[0049] Further, the biochip filter of the present invention includes

The silicon thin film layer and a part of the silicon substrate are etched and removed from the upper and lower surfaces through the SiO layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. It is characterized by being a biochip filter obtained by forming a formed filter.

[0050] With this configuration, the silicon thin film layer and the silicon substrate are etched away from the upper and lower surfaces through the SiO layer, thereby forming the well and the bottom surface of the well, and a plurality of penetrating fine cells. A hole is formed, and a filter made of silicon can be formed.

[0051] Accordingly, it is easy to provide a filter formed from silicon without complicated processes. The obtained biochip filter can be manufactured, and in the manufacturing process, the filter is not damaged and the yield is improved, and the cost can be reduced.

[0052] In addition, the biochip filter of the present invention includes the SiO layer in the well pattern portion.

Etching is removed.

[0053] With this configuration, it is possible to accurately and efficiently form a well corresponding to the well pattern.

[0054] The biochip filter of the present invention is characterized in that the surface of the silicon thin film layer in the well portion is oxidized.

[0055] Further, the biochip filter of the present invention is characterized in that the aspect ratio S of the depth with respect to the pore diameter of the pore of the filter is 5 or less.

[0056] Thus, if the aspect ratio of the depth to the pore diameter of the filter is 5 or less,

Further, pores having a uniform pore diameter can be formed during etching, and the ease of liquid flow can be maintained at a level where there is no practical problem.

[0057] Further, the biochip filter of the present invention is characterized in that the distance force between the wells is 0.05 mm or more.

[0058] Thus, if the distance force between the wells is 0.05 mm or more, sufficient tip strength can be achieved.

[0059] In addition, a biochip of the present invention is characterized by including any one of the biochip filters described above.

[0060] Further, the biochip of the present invention is characterized by including a biochip filter obtained by any one of the aforementioned biochip filters.

The invention's effect

[0061] According to the present invention, since the filter force S is formed from a metal such as silicon, for example, the strength and durability of a very thin filter having a thickness of about 1 to 10 m, for example, For example, this is a marked improvement over a biochip filter having a conventional filter composed of a metal oxide layer such as alumina, silica, and titania.

[0062] Therefore, the biochip filter of the present invention reacts or interacts with, for example, particles carrying a bioprobe such as a nucleic acid or protein and a target substance in a specimen. Separation and target washing B / F separation, and detection / identification by optical means of reaction or interaction between the target substance and bioprobe in the sample. When applied to biochip filters used in biochips, the detection and identification of these B / F separations and the optical means of the reaction or interaction between the target substance and the bioprobe in the sample are possible. It can be done reliably.

[0063] Also, when manufacturing a biochip filter, since the strength of the filter is high, the yield without breaking and damaging a part of the filter can be improved, and the cost can be reduced.

[0064] Furthermore, a biochip filter including a filter formed of a metal such as silicon can be easily manufactured without a complicated process, and the filter can be manufactured even in the manufacturing process. There is no breakage and damage, and the yield can be improved and the cost can be reduced.

Brief Description of Drawings

[0065] FIG. 1 is a top view of the biochip filter of the present invention.

[FIG. 2] FIG. 2 is a cross-sectional view taken along the A-A spring in FIG.

[Fig. 3] Fig. 3 is an enlarged plan view of one well.

[FIG. 4] FIG. 4 is a partially enlarged sectional view of one well.

FIG. 5 is a process schematic diagram showing a method for producing a biochip filter according to the present invention. FIG. 6 is a process schematic diagram showing a method for producing a biochip filter according to the present invention. FIG. 7 is a process schematic diagram showing a method for producing a biochip filter of the present invention. [FIG. 8] FIG. 8 is a process schematic diagram showing a conventional method for producing a biochip filter.

FIG. 9 is a process schematic diagram showing a conventional method for producing a biochip filter.

FIG. 10 is a process schematic diagram showing a conventional method for producing a biochip filter. [FIG. 11] FIG. 11 is a schematic diagram showing a conventional (^ 1 manufacturing method.

Explanation of symbols

10 Biochip chip

12 Filter body

14 Uenore

16 pores

18a, 18b surface

20 Silicon substrate

21 SiO layer

2

22 Silicon thin film layer

23 Ueha

24 resist film

25 recess

26 Recess

28 resist film

29 Recess

34 Dicing tape

100 silicon substrate

101 silicon substrate body

102 thermal oxide film

104 thermal oxide film

106 resist film

108 recess

110 recess

112 resist film 116 recess

124 uel

128 dicing tape

130 Biochip filter

136 filters

BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, embodiments (examples) of the present invention will be described in more detail with reference to the drawings.

[0068] FIG. 1 is a top view of the biochip filter of the present invention, FIG. 2 is a cross-sectional view taken along the line AA of FIG. 1, FIG. 3 is an enlarged plan view of one well, and FIG. FIG. 3 is a partial enlarged cross-sectional view of one well.

[0069] In Figs. 1 and 2, reference numeral 10 denotes the biochip filter of the present invention as a whole.

[0070] The biochip filter 10 of the present invention comprises, for example, a step of reacting or interacting with a particle carrying a bioprobe such as a nucleic acid or protein and a target substance in a specimen, and a step of washing the particle. These filters are used in biochips for the purpose of B / F separation of target substances containing, and detection / identification by optical means of reaction or interaction between target substances and bioprobes in the specimen.

[0071] As shown in Figs. 1 and 2, the biochip filter 10 of the present invention has a rectangular flat plate shape and includes a filter body 12 made of a metal substrate constituting the substrate portion.

[0072] The filter main body 12 is formed with a plurality of wells 14 that form cylindrical recesses that form a plurality of recesses, spaced apart from each other at a central portion.

In addition, as shown in FIGS. 3 and 4, the one well 14 is provided with a filter 18 that constitutes the bottom surface of the well 14 and has a plurality of through-holes 16 formed therein. .

[0074] In the pore 16, the probe-carrying particles that have reacted with the analyte are separated depending on the size of the pore.

In this case, in the embodiment of FIGS. 3 and 4, the pores 16 are formed to have a constant arrangement pattern at uniform intervals. The pores 16 are formed so as to be straight cylindrical pores having a uniform pore diameter. In this case, “uniform hole diameter” means that the hole diameter error is 20% or less, preferably 10% or less in terms of CV (Coefficient of Variation). A pore whose pore diameter error is 20% or less in terms of CV value can be produced by the method described later, and the dimensional difference between the probe-carrying particles contained in the well 14 and the pore diameter can be made small.

[0077] In such a filter having a uniform pore diameter, the pore diameter of the pore is not particularly limited. Usually, it is 0.Ol ^ m-lOO ^ m, preferably 0.1. ^ 111-20 ^ 111, more preferably 0.5 μm to 10 μm.

“Uniform hole interval” means that the error of the hole interval is 15% or less in terms of CV (Coefficient of Variation) value. There is no particular limitation on the pore spacing, but if the pore spacing is too narrow, the strength of the filter will be weak, and if the pore spacing is too wide, the aperture ratio will decrease, so it is usually less than twice the pore diameter, and 0 5 111 to 10 111 is preferable. The “hole interval” is the shortest distance between holes adjacent to each other.

[0079] Further, "straight" pores mean that the pores are formed without branching along the way.

[0080] For example, from the center of the opening formed on the surface 18a of one filter 18 to the other filter surface and the center of the opening formed on the surface 18b of the other filter, It is preferable that the pores are not substantially deviated from the perpendicular to the surface 18a, and the pore size on the primary side (upper surface side) of the filter 18 is different from the pore size on the secondary side (lower surface side)! / It can be something! /

[0081] The shape of the pore 16 is not particularly limited, and may be any shape such as a through-hole having a shape such as a cylinder, a quadrangular pyramid, or a polygonal pyramid. From the viewpoint of minimization, it is desirable that the cross-sectional shape of the pores perpendicular to the penetration direction of the pores is obtuse or circular.

[0082] However, when the volume of the well 14 is a predetermined amount or more, for example, 0.1 microliter or more, the meniscus is not so much a problem, and it is a through-hole having a shape such as a quadrangular column or a pyramid. There is no problem.

[0083] By making the straight pores 16 in this way, the pore length forming the filter 18 is reduced. Since it is minimized, the contact area with the pore wall is reduced, and the force S that minimizes the pumping resistance associated with filtration is reduced.

Furthermore, it is preferable that the aspect ratio of the depth to the pore diameter (diameter) of the pores 16 of the filter 18 is 5 or less.

[0085] As described above, when the aspect ratio of the depth to the pore diameter of the pores 16 of the filter 18 is 5 or less, pores having a uniform pore diameter can be formed at the time of etching. It is possible to secure a low flow rate of the liquid.

That is, as will be described later, the pores 16 of the filter 18 are formed by etching, but if the aspect ratio is large, the shape of the pores 16 may be uneven due to the etching. is there. Considering this, the aspect ratio obtained by normal chemical etching is 5 or less, and preferably 3 or less.

[0087] Further, even when the probe-carrying particles accumulate on the primary side of the filter 18 and become clogged, the particles do not enter the filter 18 and remain on the surface of the filter 18. (It becomes a so-called complete occlusion model). In this case, by flowing liquid from the secondary side of the filter 18 (referred to as “flushing”), the force S can be easily separated and dispersed from the surface of the filter 18 to the primary side.

[0088] For this reason, the clogging of the filter 18 can be resolved again and regenerated, and the life S of the filter 18 can be extended.

[0089] As described above, by forming straight pores having a uniform pore diameter in the filter 18 with uniform pore spacing, the probe-carrying particles are arranged on the primary side opening, and if necessary, It is also possible to flush the liquid such as the analyte from the secondary side and disperse the particles to the primary side, and the reaction and detection between the analyte and the probe-carrying particles can be easily performed.

[0090] Further, when the probe-carrying particles are not allowed to be discharged to the secondary side of the filter 18 and it is necessary to capture 100% of the particles, the pore size of the filter 18 is made smaller than the minimum particle size. Thus, by taking into account the particle size, 100% particle capture is possible without reducing the permeation coefficient and filtration efficiency.

[0091] The thickness of the filter 18 is preferably 1—, more preferably 2 to 7 μm. is there. When the thickness force S of the filter 18 is larger than 10 m, the filtration resistance increases when the liquid is filtered, and when the thickness of the filter 18 is less than 1 m, the mechanical strength of the filter 18 is insufficient.

[0092] Further, the aperture ratio of the filter 18 is preferably 15-60%, more preferably 20-50.

%. When the aperture ratio is less than 15%, the filtration efficiency decreases, and when the aperture ratio is greater than 60%, the mechanical strength of the filter 18 is insufficient.

[0093] On the other hand, as shown in FIGS. 3 and 4, the periphery of the filter 18 has a portion where the pores 16 are not formed, and is connected to the filter body 12. This is preferable.

[0094] Further, in this example, the diameter and height of the well 14 can be arbitrarily set according to the number of required particles and the reaction volume without particular limitation. For example, a cylindrical hole with a well diameter of 0.5 mm and a height of about 0.3 mm.

[0095] The distance force between the wels 14 is 0.05 mm or more, preferably 0.1 mm or more.

[0096] If there is a distance between the wells 14 within such a range, the chip strength that can withstand actual use can be realized.

[0097] Further, the shape of the wel 14 is not particularly limited. For example, the shape of the concave portion such as a cylinder, a quadrangular pyramid, or a polygonal pyramid may be used, but in order to minimize the meniscus. It is desirable that the cross-sectional shape of the pore perpendicular to the penetration direction of the wel 14 is obtuse or circular.

[0098] Further, in this embodiment, the force as the biochip filter 10 having a plurality of wells 14 can of course be used as the biochip filter 10 having a single well 14. The number and arrangement shape are not particularly limited.

[0099] Further, in the biochip filter 10 having a plurality of wells 14, the diameters of these wells 14 are not necessarily the same, and are different from each other in consideration of the number of contained particles and the reaction volume. It is also possible to have a uel 14.

[0100] The filter 18 of the biochip filter 10 of the present invention is made of metal. In this case, for example, silicon (Si) or the like can be used as the metal. [0101] In this case, in order to form the filter 18 from a metal, a wafer comprising a metal substrate, a metal oxide layer formed on the upper surface of the metal substrate, and a metal thin film layer formed on the upper surface of the metal oxide layer. May be used. Then, the metal thin film layer and the metal substrate are etched away from the upper and lower surfaces through the metal oxide layer, thereby forming the well 14 and the bottom surface of the well 14, and a plurality of through-holes 16 are formed. The filter 18 made of metal may be formed.

[0102] With this configuration, the metal thin film layer and the metal substrate are etched and removed from the upper and lower surfaces through the metal oxide layer, thereby forming the well and the bottom surface of the well, and a plurality of through holes are formed. Fine pores are formed, and a filter made of metal can be formed.

[0103] Therefore, it is possible to easily manufacture a biochip filter having a filter formed of metal without being a complicated process, and in the manufacturing process, there is no damage to the filter and the yield is low. Can be improved and the cost can be reduced.

[0104] For example, in the case of silicon, as will be described later, an SOI (Silicon On Insulator) comprising a silicon substrate, a SiO layer formed on the upper surface of the silicon substrate, and a silicon thin film layer formed on the upper surface of the SiO layer. ) Use a wafer. Then, by etching and removing the silicon thin film layer and the silicon substrate from the upper and lower surfaces through the SiO layer, a well 14 and a bottom surface of the well 14 are formed, and a plurality of through holes 16 are formed. A filter 18 made of silicon may be formed.

[0105] In this case, the SOI (Silicon On Insulator) wafer forms a thermal oxide film on the surface of the single crystal silicon constituting the two substrates, and the two Si substrates are bonded together via the thermal oxide film. By etching the Si substrate to a desired thickness from one side, an SOI substrate structure can be obtained (an SOI substrate manufacturing method using a bonding method).

[0106] On the other hand, the filter body 12 is formed of the same material as the filter 18 because it is formed at the same time as described later when the tool 14 and the filter 18 are produced as described above. Is. However, the filter body 12 is not limited to the one made of the same material as the filter 18.

[0107] With this configuration, the filter 18 force S is formed from a metal such as silicon, for example, so that the strength of a very thin filter having a thickness of, for example, about 1 to 10 m. The degree of durability and durability will be significantly improved compared to a biochip filter having a conventional filter composed of a metal oxide layer such as alumina, silica, or titania.

[0108] Therefore, the biochip filter 10 of the present invention comprises, for example, a step of reacting or interacting a particle carrying a nanoprobe such as a nucleic acid or a protein with a target substance in a specimen, and a particle. Used in biochips for B / F separation of target substances including washing steps, and detection and identification by optical means of reaction or interaction between target substances and bioprobes in the specimen When applied to biochip filters, these B / F separation and detection / identification by optical means such as reaction or interaction between target substance and bioprobe in the sample can be reliably performed. .

[0109] Further, when the biochip filter 10 is manufactured, the strength of the filter 18 is large, so that the yield without breaking and damaging a part of the filter is improved and the cost S can be reduced.

[0110] Hereinafter, the method for producing the biochip filter 10 of the present invention configured as described above will be described in detail with reference to the drawings, taking as an example the case of 18 filters of S and silicon.

First, as shown in FIG. 5 (A), from a silicon substrate 20, an SiO layer 21 formed on the upper surface of the silicon substrate 20, and a silicon thin film layer 22 formed on the upper surface of the SiO layer 21. Prepare the SOI (Si licon On Insulator) wafer 23.

[0112] In this case, the SOI (Silicon On Insulator) wafer forms a thermal oxide film on the surface of the single crystal silicon substrate constituting the substrate, and bonds the two Si substrates through the thermal oxide film. Use SOI substrate structure by etching Si substrate to desired thickness from one side

[0113] In this case, the thickness of the silicon substrate 20 constitutes most of the depth of the well 14, and is preferably about 300 m, for example. Further, since the silicon thin film layer 22 constitutes the filter 18, it is preferably 1-20 ^ 111, more preferably 2-5 μm. Furthermore, the SiO layer 21 functions as a protective film such as an etching stop film during the manufacturing process, and the thickness thereof is preferably 0.5 111 to 10 111. More preferably, it is 1 to 111 to 2 to 111.

Then, as shown in FIG. 5B, on the upper surface of the silicon thin film layer 22 of the SOI wafer 23.

According to photolithography, the concave pattern 2 corresponding to the hole of the filter 18 2

A resist film 24 having 5 is formed (recess forming step).

[0115] Next, as shown in FIG. 5C, the silicon thin film layer 22 is etched away using the resist layer 24 as a mask, so that the silicon thin film layer 22 corresponds to the pores of the filter. A recess 26 is formed.

In this case, as the etching, a reactive ion etching method (RIE method) using a fluorine-based gas may be used. At this time, since the SiO layer 21 is not etched by dry etching, a recess 26 having a predetermined depth corresponding to the thickness of the Si layer 22 is formed.

Then, as shown in FIG. 5D, the resist film 24 is peeled off.

[0118] Then, with the top and bottom reversed, as shown in FIG. 6A, a resist film 28 is formed on the upper surface of the silicon substrate 20 of the SOI wafer 23. A resist mask layer 28 having a recess 29 pattern corresponding to the well 14 is formed.

[0119] Then, as shown in FIG. 6B, the exposed portion of the silicon substrate 20 is etched away using the resist film 28 as a mask to form the well 14 (well forming step). ).

In this case, as the silicon etching, a dry etching method (RIE method) using a fluorine-based gas may be used. At this time, since the SiO layer 21 is not etched by dry etching, the well 14 having a predetermined depth is formed.

Then, as shown in FIG. 6C, the SiO layer 21 is removed by etching.

[0122] Next, as shown in FIG. 7A, after dicing tape 34 is attached to the lower surface of the silicon substrate 20, dicing is performed as shown in FIG. 7B. Divide into individual biochip filters 10.

[0123] In this case, dicing using a dicing cutter or the like can be used as dicing.

[0124] Then, as shown in FIG. 7C, the dicing tape 34 is expanded while the temperature is increased. By pulling outward, the dicing tape 34 and individual chips are peeled off and separated into individual biochip filters 10.

Accordingly, as shown in FIG. 7 (D), the filter main body 12 made of the silicon substrate 20 constituting the substrate portion and the well 14 forming a plurality of recesses provided in the filter main body 12. Thus, the biochip filter 10 provided with the filter 18 made of silicon that forms the bottom surface of the well 14 and has a plurality of through-holes 16 formed therein is obtained.

In FIGS. 5 to 7, for convenience of explanation, the force indicating one well 14 is actually formed with a plurality of wells 14 spaced apart from each other.

[0127] Further, it is preferable that the surface force of the silicon thin film layer 22 in the portion of the well 14 is oxidized. Thus, if the surface force of the silicon thin film layer 22 in the portion of the well 14 is oxidized, foreign matters such as particles adhering to the surface of the silicon thin film layer 22 can be easily lifted off with a solution such as diluted hydrofluoric acid. it can. As this oxidation method, a thermal oxidation method can be used.

[0128] While the preferred embodiments of the present invention have been described above, the present invention is not limited thereto. For example, in the above embodiment, an SOI (Silicon On Insulat or SOI) wafer using a bonding method is used. Various examples can be used without departing from the object of the present invention, such as the use of the SOI wafer formed by the SIMOX method.

Yes

Claims

The scope of the claims

[1] A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

The metal thin film layer and a part of the metal substrate are etched and removed from the upper and lower surfaces through the metal oxide layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. A filter for biochip, characterized by forming a filter formed from

[2] Using a wafer comprising the metal substrate, a metal oxide layer formed on the upper surface of the metal substrate, and a metal thin film layer formed on the upper surface of the metal oxide layer,

The method for producing a biochip filter according to claim 1, comprising:

[3] A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

The filter is formed of metal; The metal substrate force constituting the filter body is composed of a silicon substrate,

The silicon thin film layer and a part of the silicon substrate are etched and removed from the upper and lower surfaces through the SiO layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. A method for manufacturing a biochip filter, comprising: forming a filter.

[4] Using an SOI (Silicon On Insulator) wafer comprising the silicon substrate, a SiO layer formed on the upper surface of the silicon substrate, and a silicon thin film layer formed on the upper surface of the SiO layer, the silicon thin film of the SOI wafer A recess forming step of forming a recess to form a pore of the filter by etching away a part of the layer;

The method for producing a biochip filter according to claim 3, comprising:

[5] The method for producing a biochip filter according to any one of claims 3 to 4, wherein the SiO layer force in the well pattern portion is removed by etching.

[6] The method for producing a biochip filter according to any one of claims 3 to 5, wherein a surface force of the silicon thin film layer in the well portion is oxidized.

[7] The method for producing a biochip filter according to any one of [1] to [6], wherein a specific force of a filter thickness to a pore diameter of the filter is 5 or less.

Yes

[8] The distance force between the wels is 0.05 mm or more. 8. A method for producing a biochip filter according to any one of 7 above.

A biochip comprising the biochip filter obtained by the method for producing a biochip filter according to claim 1.

A filter body made of a metal substrate constituting the substrate part;

A well forming a recess provided in the filter body,

A filter chip for biochip, characterized in that the filter is made of S and a metal.

11. The biochip filter according to claim 10, wherein the filter is made of metallic silicon that constitutes the filter.

The silicon thin film layer and a part of the silicon substrate are etched and removed from the upper and lower surfaces through the SiO layer to form a well and a bottom surface of the well, and a plurality of through-holes are formed. 12. The biochip filter according to claim 11, which is a biochip filter obtained by forming a formed filter.

13. The biochip filter according to claim 12, wherein the SiO layer force in the well pattern portion is removed by etching.

14. The biochip filter according to any one of claims 11 and 13, wherein a surface force of the silicon thin film layer in the well portion is oxidized.

15. The biochip filter according to claim 10, wherein an aspect ratio of a depth to a pore diameter of the filter is 5 or less.

The biochip filter according to any one of claims 10 to 15, wherein a distance force between the wells is 0.05 mm or more. [17] A biochip comprising the biochip filter according to any one of claims 10 to 16.