U.S. patent application number 14/386527 was filed with the patent office on 2015-02-19 for chip for analysis of target substance.
The applicant listed for this patent is NEC Corporation. Invention is credited to Minoru Asogawa, Hisashi Hagiwara, Yasuo Iimura, Yoshinori Mishina.
Application Number | 20150050721 14/386527 |
Document ID | / |
Family ID | 49222314 |
Filed Date | 2015-02-19 |
United States Patent
Application |
20150050721 |
Kind Code |
A1 |
Asogawa; Minoru ; et
al. |
February 19, 2015 |
CHIP FOR ANALYSIS OF TARGET SUBSTANCE
Abstract
The present invention provides a chip for analysis of a target
substance that is compact and allows analysis of a target substance
with less time and effort. The chip for analysis of a target
substance includes a first flexible substrate 1, a second flexible
substrate 2, and a third substrate 3. A flow channel-forming
non-bonded area 11 is formed on a bonding surface of the first
flexible substrate 1 and the second flexible substrate 2 in a
band-like manner and an extraction chamber-forming non-bonded area
5 having a wider band width than the flow channel-forming
non-bonded area 11 is formed at a part of the flow channel-forming
non-bonded area 11. The first flexible substrate 1 includes a
through-hole 7 that is in contact with the flow channel-forming
non-bonded area 11, a shutter-forming non-bonded area 12b is formed
on a bonding surface of the second flexible substrate 2 and the
third substrate 3 at a far side of the through-hole 7 relative to
the extraction chamber-forming non-bonded area 5 such that the
shutter-forming non-bonded area 12b and the flow channel-forming
non-bonded area 11 intersect above and below, the first flexible
substrate 1 and the second flexible substrate 2 include a pressure
supply port 18b that comes through the substrates so as to be in
contact with the shutter-forming non-bonded area 12b, and a
magnetic particle 16 that binds to a target substance is placed
above the extraction chamber-forming non-bonded area 5.
Inventors: |
Asogawa; Minoru; (Tokyo,
JP) ; Hagiwara; Hisashi; (Tokyo, JP) ;
Mishina; Yoshinori; (Tokyo, JP) ; Iimura; Yasuo;
(Tokyo, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NEC Corporation |
Tokyo |
|
JP |
|
|
Family ID: |
49222314 |
Appl. No.: |
14/386527 |
Filed: |
January 23, 2013 |
PCT Filed: |
January 23, 2013 |
PCT NO: |
PCT/JP2013/051332 |
371 Date: |
September 19, 2014 |
Current U.S.
Class: |
435/287.2 ;
204/603; 422/503 |
Current CPC
Class: |
C12Q 1/686 20130101;
B01L 2400/0481 20130101; B01L 2300/0816 20130101; B01L 2300/123
20130101; B01L 2400/043 20130101; G01N 27/453 20130101; B01L
3/502738 20130101; B01L 2200/0668 20130101; B01L 3/502707 20130101;
B01L 3/502715 20130101; B01L 2300/0867 20130101; G01N 27/745
20130101; B01L 2400/0655 20130101; G01N 27/44791 20130101; B01L
2300/0861 20130101; B01L 3/502761 20130101; G01N 27/44704 20130101;
B01L 2300/0864 20130101; B01L 2400/0487 20130101; B01F 11/0051
20130101; B01L 2300/0645 20130101; B01L 2300/0874 20130101; B01L
2300/0887 20130101 |
Class at
Publication: |
435/287.2 ;
422/503; 204/603 |
International
Class: |
B01L 3/00 20060101
B01L003/00; G01N 27/447 20060101 G01N027/447 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 21, 2012 |
JP |
2012-063645 |
Claims
1. A chip for analysis of a target substance comprising: a
laminate, the laminate comprising: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner and an extraction chamber-forming non-bonded area
having a wider band width than the flow channel-forming non-bonded
area is formed at a part of the flow channel-forming non-bonded
area, the first flexible substrate comprises a through-hole that is
in contact with the flow channel-forming non-bonded area, a
shutter-forming non-bonded area is formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner at a far side of the through-hole relative to the
extraction chamber-forming non-bonded area such that the
shutter-forming non-bonded area and the flow channel-forming
non-bonded area intersect above and below via the second flexible
substrate being interposed therebetween, at least one of both of
the first flexible substrate and the second flexible substrate and
the third substrate comprises a pressure supply port that comes
through the substrate(s) so as to be in contact with the
shutter-forming non-bonded area, a magnetic particle that binds to
a target substance is placed above the extraction chamber-forming
non-bonded area, a flow channel and an extraction chamber are
formable by supplying pressure from the through-hole to raise a
site above the flow channel-forming non-bonded area and a site
above the extraction chamber-forming non-bonded area, the flow
channel is blockable by supplying pressure from the pressure supply
port to raise a site above the shutter-forming non-bonded area, and
the target substance that is bound to the magnetic particle is
capturable by generating a magnetic field at at least one of an
undersurface of the third substrate directly below an end of the
extraction chamber at the opposite side of the through-hole and a
top surface of the first flexible substrate directly above an end
of the extraction chamber at the opposite side of the
through-hole.
2. A chip for analysis of a target substance comprising: a
laminate, the laminate comprising: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner and a mixing chamber-forming non-bonded area
having a wider band width than the flow channel-forming non-bonded
area is formed at a part of the flow channel-forming non-bonded
area, the first flexible substrate comprises a through-hole that is
in contact with the flow channel-forming non-bonded area,
shutter-forming non-bonded areas are formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner at a near side and a far side of the through-hole
relative to the mixing chamber-forming non-bonded area such that
the shutter-forming non-bonded areas and the flow channel-forming
non-bonded area intersect above and below via the second flexible
substrate being interposed therebetween, at least one of both of
the first flexible substrate and the second flexible substrate and
the third substrate comprises pressure supply ports that come
through the substrate(s) so as to be in contact with the
shutter-forming non-bonded areas, a flow channel and a mixing
chamber are formable by supplying pressure from the through-hole to
raise a site above the flow channel-forming non-bonded area and a
site above the mixing chamber-forming non-bonded area, the flow
channel is blockable by supplying pressure from the pressure supply
ports to raise sites above the shutter-forming non-bonded areas,
and a target substance and a reagent are mixable in the mixing
chamber by applying pressure to a top surface of the first flexible
substrate above the mixing chamber to deform the mixing
chamber.
3. A chip for analysis of a target substance comprising: a
laminate, the laminate comprising: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner, the first flexible substrate comprises a
through-hole that is in contact with the flow channel-forming
non-bonded area, a first mixing chamber-forming non-bonded area and
a second mixing chamber-forming non-bonded area each having a wider
band width than the flow channel-forming non-bonded area are each
formed at a part of the flow channel-forming non-bonded area on the
bonding surface of the first flexible substrate and the second
flexible substrate in this order from the through-hole side,
shutter-forming non-bonded areas are formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner at a near side of the through-hole relative to the
first mixing chamber-forming non-bonded area and a far side of the
through-hole relative to the second mixing chamber-forming
non-bonded area such that the shutter-forming non-bonded areas and
the flow channel-forming non-bonded area intersect above and below
via the second flexible substrate being interposed therebetween, at
least one of both of the first flexible substrate and the second
flexible substrate and the third substrate includes pressure supply
ports that come through the substrate(s) so as to be in contact
with the shutter-forming non-bonded areas, a flow channel, a first
mixing chamber, and a second mixing chamber are formable by
supplying pressure from the through-hole to raise a site above the
flow channel-forming non-bonded area, a site above the first mixing
chamber-forming non-bonded area, and a site above the second mixing
chamber-forming non-bonded area, the flow channel is blockable by
supplying pressure from the pressure supply ports to raise sites
above the shutter-forming non-bonded areas, and a target substance
and a reagent are mixable by moving them between the first mixing
chamber and the second mixing chamber.
4. The chip according to claim 3, wherein a shutter-forming
non-bonded area is further formed on the bonding surface of the
second flexible substrate and the third substrate in a band-like
manner at a far side of the through-hole relative to the first
mixing chamber-forming non-bonded area such that the
shutter-forming non-bonded area and the flow channel-forming
non-bonded area intersect above and below via the second flexible
substrate being interposed therebetween.
5. The chip according to any one of claims 1 to 4, further
comprising: an amplification portion.
6. The chip according to claim 5, wherein the amplification portion
comprises a PCR amplification portion.
7. The chip according to any one of claims 1 to 6, further
comprising: an analysis portion.
8. The chip according to claim 7, wherein the analysis portion
comprises an electrophoresis analysis portion.
Description
TECHNICAL FIELD
[0001] The present invention relates to a chip for analysis of a
target substance.
BACKGROUND ART
[0002] Conventionally, various kinds of DNA analyzers have been
proposed (see, for example, Patent Document 1). Conventional DNA
analyzers have been large analyzers in which reaction vessels,
photodetectors, amplifiers, and the like are independently
provided, respectively, and therefore have required wide
installation spaces. Furthermore, conventional DNA analyzers have
required great deal of time and efforts.
CITATION LIST
Patent Document(s)
Patent Document 1: JP 2009-247297 A
SUMMARY OF INVENTION
Problem to be Solved by the Invention
[0003] Hence, the present invention is intended to provide a chip
for analysis of a target substance that is compact and allows
analysis of a target substance such as DNA with less time and
effort.
Means for Solving Problem
[0004] In order to achieve the above object, the present invention
provides a first chip for analysis of a target substance,
including:
a laminate, the laminate including: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner and an extraction chamber-forming non-bonded area
having a wider band width than the flow channel-forming non-bonded
area is formed at a part of the flow channel-forming non-bonded
area, the first flexible substrate includes a through-hole that is
in contact with the flow channel-forming non-bonded area, a
shutter-forming non-bonded area is formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner at a far side of the through-hole relative to the
extraction chamber-forming non-bonded area such that the
shutter-forming non-bonded area and the flow channel-forming
non-bonded area intersect above and below via the second flexible
substrate being interposed therebetween, at least one of both of
the first flexible substrate and the second flexible substrate and
the third substrate includes a pressure supply port that comes
through the substrate(s) so as to be in contact with the
shutter-forming non-bonded area, a magnetic particle that binds to
a target substance is placed above the extraction chamber-forming
non-bonded area, a flow channel and an extraction chamber are
formable by supplying pressure from the through-hole to raise a
site above the flow channel-forming non-bonded area and a site
above the extraction chamber-forming non-bonded area, the flow
channel is blockable by supplying pressure from the pressure supply
port to raise a site above the shutter-forming non-bonded area, and
the target substance that is bound to the magnetic particle is
capturable by generating a magnetic field at at least one of an
undersurface of the third substrate directly below an end of the
extraction chamber at the opposite side of the through-hole and a
top surface of the first flexible substrate directly above an end
of the extraction chamber at the opposite side of the
through-hole.
[0005] The present invention also provides a second chip for
analysis of a target substance, including:
a laminate, the laminate including: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner and a mixing chamber-forming non-bonded area
having a wider band width than the flow channel-forming non-bonded
area is formed at a part of the flow channel-forming non-bonded
area, the first flexible substrate includes a through-hole that is
in contact with the flow channel-forming non-bonded area,
shutter-forming non-bonded areas are formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner (e.g. two bands-like manner) at a near side and a
far side of the through-hole relative to the mixing chamber-forming
non-bonded area such that the shutter-forming non-bonded areas and
the flow channel-forming non-bonded area intersect above and below
via the second flexible substrate being interposed therebetween, at
least one of both of the first flexible substrate and the second
flexible substrate and the third substrate includes pressure supply
ports that come through the substrate(s) so as to be in contact
with the shutter-forming non-bonded areas, a flow channel and a
mixing chamber are formable by supplying pressure from the
through-hole to raise a site above the flow channel-forming
non-bonded area and a site above the mixing chamber-forming
non-bonded area, the flow channel is blockable by supplying
pressure from the pressure supply ports to raise sites above the
shutter-forming non-bonded areas, and a target substance and a
reagent are mixable in the mixing chamber by applying pressure to a
top surface of the first flexible substrate above the mixing
chamber to deform the mixing chamber.
[0006] The present invention also provides a third chip for
analysis of a target substance, including:
a laminate, the laminate including: a first flexible substrate; a
second flexible substrate; and a third substrate, wherein a flow
channel-forming non-bonded area is formed on a bonding surface of
the first flexible substrate and the second flexible substrate in a
band-like manner, the first flexible substrate includes a
through-hole that is in contact with the flow channel-forming
non-bonded area, a first mixing chamber-forming non-bonded area and
a second mixing chamber-forming non-bonded area each having a wider
band width than the flow channel-forming non-bonded area are each
formed on the bonding surface of the first flexible substrate and
the second flexible substrate at a part of the flow channel-forming
non-bonded area in this order from the through-hole side,
shutter-forming non-bonded areas are formed on a bonding surface of
the second flexible substrate and the third substrate in a
band-like manner (e.g. two bands-like manner) at a near side of the
through-hole relative to the first mixing chamber-forming
non-bonded area and a far side of the through-hole relative to the
second mixing chamber-forming non-bonded area such that the
shutter-forming non-bonded areas and the flow channel-forming
non-bonded area intersect above and below via the second flexible
substrate being interposed therebetween, at least one of both of
the first flexible substrate and the second flexible substrate and
the third substrate includes pressure supply ports that come
through the substrate(s) so as to be in contact with the
shutter-forming non-bonded areas, a flow channel, a first mixing
chamber, and a second mixing chamber are formable by supplying
pressure from the through-hole to raise a site above the flow
channel-forming non-bonded area, a site above the first mixing
chamber-forming non-bonded area, and a site above the second mixing
chamber-forming non-bonded area, the flow channel is blockable by
supplying pressure from the pressure supply ports to raise sites
above the shutter-forming non-bonded areas, and a target substance
and a reagent are mixable by moving them between the first mixing
chamber and the second mixing chamber.
Effects of the Invention
[0007] According to the present invention, it is possible to
provide a chip for analysis of a target substance that is compact
and allows analysis of a target substance such as DNA with less
time and effort.
BRIEF DESCRIPTION OF DRAWINGS
[0008] FIG. 1 shows diagrams illustrating an example of the
configuration of the first chip for analysis of a target substance
of the present invention; FIG. 1(A) is a schematic perspective plan
view; FIG. 1(B) is a schematic cross sectional view of FIG. 1(A)
viewed from the line I-I; and FIG. 1(C) is a schematic cross
sectional view of FIG. 1(A) viewed from the line II-II.
[0009] FIG. 2 shows schematic cross sectional views illustrating an
example of the usage of the chip for analysis of a target substance
shown in FIG. 1.
[0010] FIG. 3 shows schematic cross sectional views illustrating an
example of the configuration of the second chip for analysis of a
target substance of the present invention.
[0011] FIG. 4 shows schematic cross sectional views illustrating an
example of the configuration of the third chip for analysis of a
target substance of the present invention.
[0012] FIG. 5 is a schematic perspective plan view showing another
example of the configuration of the chip for analysis of a target
substance of the present invention.
DESCRIPTION OF EXEMPLARY EMBODIMENT
[0013] The chip for analysis of a target substance of the present
invention will be described with reference to exemplary
embodiments. Note here that the present invention is not limited to
these exemplary embodiments. Furthermore, the description for each
Embodiment can be applied to another Embodiment unless otherwise
indicated.
Embodiment 1
[0014] FIG. 1 shows an example of the configuration of the first
chip for analysis of a target substance of the present invention.
In FIG. 1, FIG. 1(A) is a schematic perspective plan view, FIG.
1(B) is a schematic cross sectional view of FIG. 1(A) viewed from
the line I-I, and FIG. 1(C) is a schematic cross sectional view of
FIG. 1(A) viewed from the line II-II. As shown in FIG. 1, a chip
for analysis of a target substance 10 includes a laminate in which
a first flexible substrate 1, a second flexible substrate 2, and a
third substrate 3 are laminated. In the laminate, the laminate
direction of substrates is referred to as the up-and-down direction
(hereinafter, the same applies). A flow channel-forming non-bonded
area 11 is formed on the bonding surface of the first flexible
substrate 1 and the second flexible substrate 2 in a band-like
manner and an extraction chamber-forming non-bonded area 5 having a
wider band width than the flow channel-forming non-bonded area 11
is formed at a part of the flow channel-forming non-bonded area 11.
The first flexible substrate 1 includes a through-hole 7 that is in
contact with the flow channel-forming non-bonded area 11.
Shutter-forming non-bonded areas 12a and 12b are formed on the
bonding surface of the second flexible substrate 2 and the third
substrate 3 in a band-like manner at the near side and the far side
of the through-hole 7 relative to the extraction chamber-forming
non-bonded area 5 such that the shutter-forming non-bonded areas
12a and 12b and the flow channel-forming non-bonded area 11
intersect above and below via the second flexible substrate 2 being
interposed therebetween. The first flexible substrate 1 and the
second flexible substrate 2 include pressure supply ports 18a and
18b that come through the substrates so as to be in contact with
the shutter-forming non-bonded areas 12a and 12b respectively. It
is also possible that the third substrate 3 includes the pressure
supply ports 18a and 18b that come through the substrate so as to
be in contact with the shutter-forming non-bonded areas 12a and 12b
respectively. Although it is not shown in FIG. 1, a magnetic
particle that binds to a target substance is placed above the
extraction chamber-forming non-bonded area 5. In this Embodiment,
the shutter-forming non-bonded area 12a and the pressure supply
port 18a are optional components and are not indispensable,
although the chip for analysis of a target substance of this
Embodiment preferably includes these components.
[0015] In this Embodiment, the flow direction of liquid in the flow
channel to be formed is along the flow channel-forming non-bonded
area 11 and the through-hole 7 side is the upstream side.
Therefore, it can be said that the shutter-forming non-bonded area
12a is formed at the downstream side of the through-hole 7 and the
upstream side of the extraction chamber-forming non-bonded area 5,
i.e., between the through-hole 7 and the extraction chamber-forming
non-bonded area 5, and the shutter-forming non-bonded area 12b is
formed at the downstream side of the extraction chamber-forming
non-bonded area 5.
[0016] In FIG. 1, one through-hole 7 is provided at the left end of
the flow channel-forming non-bonded area 11. However, the present
invention is not limited thereto. An appropriate number of
through-holes may be provided at any place as long as the
through-hole is in contact with the flow channel-forming non-bonded
area 11.
[0017] Furthermore, in FIG. 1, one pressure supply port 18a is
provided at the end of the shutter-forming non-bonded area 12a and
one pressure supply port 18b is provided at the end of the
shutter-forming non-bonded area 12b. However, the present invention
is not limited thereto. An appropriate number of pressure supply
ports may be provided at any place as long as the pressure supply
port is in contact with the shutter-forming non-bonded area.
[0018] There is no particular limitation on how the shutter-forming
non-bonded areas 12a and 12b and the flow channel-forming
non-bonded area 11 intersect as long as the shutter-forming
non-bonded areas 12a and 12b and the flow channel-forming
non-bonded area 11 intersect above and below via the second
flexible substrate 2 being interposed therebetween. For example,
although the shutter-forming non-bonded areas 12a and 12b and the
flow channel-forming non-bonded area 11 intersect at right angles
in FIG. 1, the present invention is not limited thereto.
[0019] The undersurface of the first flexible substrate 1 and the
top surface of the second flexible substrate 2 are bonded with each
other at around the flow channel-forming non-bonded area 11,
through-hole 7, and the extraction chamber-forming non-bonded area
5. Preferably, the undersurface of the first flexible substrate 1
and the top surface of the second flexible substrate 2 are bonded
with each other at an area excluding the flow channel-forming
non-bonded area 11, the through-hole 7, and the extraction
chamber-forming non-bonded area 5. Furthermore, the undersurface of
the second flexible substrate 2 and the top surface of third
substrate 3 are bonded with each other at an area excluding the
shutter-forming non-bonded areas 12a and 12b and the pressure
supply ports 18a and 18b.
[0020] The chip for analysis of a target substance 10 can be
produced, for example, as follows. First, the first flexible
substrate 1, the second flexible substrate 2, and the third
substrate 3 are provided. Surface modification treatment for the
purpose of increasing the bonding strength between the substrates
may be applied to the undersurface of the first flexible substrate
1, the top surface and the undersurface of the second flexible
substrate 2, and the top surface of the third substrate 3. Examples
of the surface modification treatment include oxygen plasma
treatment and excimer UV light irradiation treatment. The oxygen
plasma treatment can be performed, for example, using a reactive
ion etching (RIE) apparatus and the like in the presence of oxygen.
The excimer UV light irradiation treatment can be performed, for
example, using a dielectric barrier discharge lamp under an air
atmosphere of atmospheric pressure.
[0021] Examples of the material of the first flexible substrate 1
include a silicone rubber such as polydimethylsiloxane (PDMS); a
nitrile rubber; a hydrogenated nitrile rubber; a fluororubber; an
ethylene-propylene rubber; a chloroprene rubber; an acrylic rubber;
a butyl rubber; an urethane rubber; a chlorosulfonated polyethylene
rubber; an epichlorohydrin rubber; a natural rubber; an isoprene
rubber; a styrene-butadiene rubber; a butadiene rubber; a
polysulfide rubber; a norbornene rubber; and a thermoplastic
elastomer. These materials may be used alone or two or more of them
may be used in combination. Among them, a silicone rubber such as
PDMS is particularly preferable. The thickness of the first
flexible substrate 1 is, for example, in the range from 10 .mu.m to
5 mm in consideration of the strength thereof and the formation of
the flow channel and the extraction chamber that will be described
below.
[0022] There are no particular limitations on the methods of
forming the through-hole 7 and the pressure supply ports 18a and
18b on the first flexible substrate 1, and conventionally known
methods can be used. There are no particular limitations on the
shapes of the through-hole 7 and the pressure supply ports 18a and
18b. The through-hole 7 and the pressure supply ports 18a and 18b
can take any shape such as a cylinder shown in FIG. 1 and a prism,
for example. The sizes of the through-hole 7 and the pressure
supply ports 18a and 18b may be set appropriately, for example,
according to the widths of the flow channel-forming non-bonded area
and the shutter-forming non-bonded area that will be described
below.
[0023] Examples of the material of the second flexible substrate 2
include those described for the first flexible substrate 1. While
the material of the second flexible substrate 2 can be the same as
or different from the material of the first flexible substrate, the
material of the second flexible substrate 2 is preferably the same
as the material of the first flexible substrate 1. Specifically,
for example, in the case where the first flexible substrate 1 is
silicone rubber, the second flexible substrate 2 is preferably
silicone rubber. If the first flexible substrate 1 and the second
flexible substrate 2 are both silicone rubber, the first flexible
substrate 1 and the second flexible substrate 2 can be bonded by a
self adsorption ability without using an adhesive agent. The
thickness of the second flexible substrate 2 is, for example, in
the range from 10 .mu.m to 500 .mu.m in consideration of the
strength thereof and the blocking of the flow channel that will be
described below.
[0024] There are no particular limitations on the methods of
forming the pressure supply ports 18a and 18b on the second
flexible substrate 2, and conventionally known methods can be used.
The shapes and the sizes of the pressure supply ports 18a and 18b
of the second flexible substrate 2 are, for example, the same as
those of the pressure supply ports 18a and 18b of the first
flexible substrate 1.
[0025] The flow channel-forming non-bonded area 11 is formed on the
top surface of the second flexible substrate 2 in a band-like
manner and the extraction chamber-forming non-bonded area 5 having
a wider band width than the flow channel-forming non-bonded area 11
is formed at a part of the flow channel-forming non-bonded area 11.
The flow channel-forming non-bonded area 11 and the extraction
chamber-forming non-bonded area 5 each can be formed as, for
example, an electrode film, a dielectric protective film, a
semiconductor film, a fluorescent film, a superconductive film, a
dielectric film, a solar cell film, an antireflection film, a
wear-resistant film, an optical interference film, a reflection
film, an antistatic film, a conductive film, an antifouling film, a
hard coating film, a barrier film, an electromagnetic wave
shielding film, an infrared shielding film, an ultraviolet
absorbing film, a lubricating film, a shape memory film, a magnetic
recording film, a light-emitting element film, a biocompatible
film, a corrosion-resistant film, a catalyst film, or a gas sensor
film, for example, by a conventionally known chemical thin film
formation technology.
[0026] Specifically, for example, the aforementioned thin film can
be formed by a plasma discharge treatment apparatus using an
organic fluorine compound or a metal compound as reactive gas.
[0027] Examples of the organic fluorine compound include
fluorocarbon compounds such as fluoromethane, fluoroethane,
tetrafluoromethane, hexafluoromethane, 1,1,2,2-tetrafluoroethylene,
1,1,1,2,3,3-hexafluoropropane, hexafluoropropene, and
6-fluoropropylen; fluorohydrocarbon compounds such as
1,1-difluoroethylene, 1,1,1,2-tetrafluoroethane, and
1,1,2,2,3-pentafluoropropane; carbon fluorochloride compounds such
as difluorodichloromethane and trifluorochloromethane;
fluoroalcohols such as 1,1,1,3,3,3-hexafluoro-2-propanol,
1,3-difluoro-2-propanol, and perfluorobutanol; fluoro carboxylic
ester such as vinyltrifluoroacetate and 1,1,1-trifluoroacetate; and
fluoroketone such as acetyl fluoride, hexafluoro acetone, and
1,1,1-trifluoroacetone.
[0028] Examples of the metal compound include a single metal
compound, a mixed metal compound, and an organic metal compound of
Al, As, Au, B, Bi, Ca, Cd, Cr, Co, Cu, Fe, Ga, Ge, Hg, In, Li, Mg,
Mn, Mo, Na, Ni, Pb, Pt, Rh, Sb, Se, Si, Sn, Ti, V, W, Y, Zn, and
Zr.
[0029] The aforementioned thin film can be formed, for example, by
a reactive ion etching system (RIE), a printing method, and the
like in the presence of fluorocarbon (CHF.sub.3) via a mask. As the
printing method, for example, conventionally known printing methods
such as roll printing, pattern printing, decalcomania, and
electrostatic copying can be employed. In the case where the
aforementioned thin film is formed by the printing method, for
example, a metal fine particle, a conductive ink, an insulating
ink, a carbon fine particle, a silane agent, parylene, a paint, a
pigment, a dye, a water-based dye ink, a water-based pigment ink,
an oil dye ink, an oil pigment ink, a solvent ink, a solid ink, a
gel ink, a polymer ink, and the like can be used suitably for the
material for forming the thin film. Examples of the metal fine
particle include a single metal fine particle of, a mixed metal
fine particle of two or more of, an oxide fine particle (for
example, ITO fine particle or the like) of the single metal or the
mixed metal of, and an organic metal compound fine particle of Al,
As, Au, B, Bi, Ca, Cd, Cr, Co, Cu, Fe, Ga, Ge, Hg, In, Li, Mg, Mn,
Mo, Na, Ni, Pb, Pt, Rh, Sb, Se, Si, Sn, Ti, V, W, Y, Zn, and
Zr.
[0030] The thickness of each of the flow channel-forming non-bonded
area 11 and the extraction chamber-forming non-bonded area 5 is,
for example, in the range from 10 nm to 10 .mu.m and preferably in
the range from 50 nm to 3 .mu.m in consideration of uniform
formation of the flow channel-forming non-bonded area 11 and the
extraction chamber-forming non-bonded area 5 and the bonding
ability between the first flexible substrate 1 and the second
flexible substrate 2 at an area excluding the non-bonded area. The
width of the flow channel-forming non-bonded area 11 is, for
example, in the range from 10 .mu.m to 3000 .mu.m in consideration
of the formation of the flow channel that will be described below,
a supply amount of each of a reagent and a target substance such as
DNA, and the like. The size of the extraction chamber-forming
non-bonded area 5 is, for example, in the range from 3 mm.sup.2 to
300 mm.sup.2 and preferably in the range from 16 mm.sup.2 to 50
mm.sup.2 in consideration of the formation of the extraction
chamber that will be described below, a supply amount of each of a
reagent and a target substance such as DNA, and the like.
[0031] The shape of the flow channel-forming non-bonded area 11 is
not limited to a linear band shown in FIG. 1, and, for example,
various shaped bands such as a Y-shaped band and an L-shaped band
can be employed. The shape of the extraction chamber-forming
non-bonded area 5 is also not limited to a circle shown in FIG. 1,
and, for example, any shape such as a rectangle can be
employed.
[0032] Examples of the material of the third substrate 3 include
acryl, a silicone rubber such as PDMS, glass, polyethylene
terephthalate, polyethylene naphthalate, polyethylene,
polypropylene, cellophane, cellulose diacetate, cellulose acetate
butyrate, cellulose acetate propionate, cellulose acetate
phthalate, cellulose triacetate, cellulose nitrate, polyvinylidene
chloride, polyvinyl alcohol, ethylene vinyl alcohol, polycarbonate,
a norbornene resin, polymethylpentene, polyether ketone, polyimide,
polyethersulfone, polyether ketone imide, polyamide, a fluororesin,
nylon, polymethyl methacrylate, polyarylate, a polylactic resin,
polybutylene succinate, a nitrile rubber, a hydrogenated nitrile
rubber, a fluororubber, an ethylene-propylene rubber, a chloroprene
rubber, an acrylic rubber, a butyl rubber, an urethane rubber, a
chlorosulfonated polyethylene rubber, an epichlorohydrin rubber, a
natural rubber, an isoprene rubber, a styrene-butadiene rubber, a
butadiene rubber, a polysulfide rubber, a norbornene rubber, and
thermoplastic elastomer. These materials may be used alone or two
or more of them may be used in combination. Among them, acryl is
particularly preferable. The thickness of the third substrate 3 is,
for example, in the range from 300 .mu.m to 10 mm in consideration
of strength and economic efficiency.
[0033] Preferably, surface treatment using a surface treatment
agent is applied to the top surface of the third substrate 3 for
the purpose of increasing the bonding ability between the top
surface of the third substrate 3 and the undersurface of the second
flexible substrate 2 at an area excluding the non-bonded area.
Examples of the surface treatment agent include alkylsilane such as
dimethylsilane, tetramethylsilane, and tetraethylsilane; organic
silicon compounds of silicon alkoxysilane such as
tetramethoxysilane, tetraethoxysilane, tetrapropoxysilane,
dimethyldiethoxysilane, methyltrimethoxysilane, and
ethyltriethoxysilane; silicon-hydrogen compounds such as monosilane
and disilane; halogenated silicon compounds such as dichlorosilane,
trichlorosilane, and tetrachlorosilane; silazane such as
hexamethyldisilazane; and silicon compounds into which functional
groups are introduced such as vinyl, epoxy, styryl, methacryloxy,
acryloxy, amino, ureide, chloropropyl, mercapto, sulfide, and
isocyanate.
[0034] Shutter-forming non-bonded areas 12a and 12b are formed on
the top surface of the third substrate 3 in a band-like manner. The
shutter-forming non-bonded areas 12a and 12b may be formed, for
example, using the same material as those used for the flow
channel-forming non-bonded area 11 and the extraction
chamber-forming non-bonded area 5 such that the shutter-forming
non-bonded areas 12a and 12b have the same thickness as the flow
channel-forming non-bonded area 11 and the extraction
chamber-forming non-bonded area 5. The width of each of the
shutter-forming non-bonded areas 12a and 12b is, for example, in
the range from 10 .mu.m to 5000 .mu.m in consideration of the
blocking of the flow channel that will be described below and
economic efficiency.
[0035] Next, the first flexible substrate 1, the second flexible
substrate 2, and the third substrate 3 are laminated. At this time,
although it is not shown, a magnetic particle that binds to a
target substance such as DNA is placed above the extraction
chamber-forming non-bonded area 5. "Bonding" may be, for example,
direct or indirect bonding of the target substance to the magnetic
particle. In the former case, an example of the direct bonding
includes adhesion of the target substance to the magnetic particle
itself. In the latter case, examples of the indirect bonding
include adsorption or adhesion of the target substance to a
predetermined substance to which a magnetic particle coating is
applied and bonding of the target substance to the magnetic
particle by reaction using a reactive substance.
[0036] Preferably, the magnetic particle is, for example, a sphere
and the particle size is, for example, in the range from 0.3 .mu.m
to 5 .mu.m. As the magnetic particle, for example, a substance
having a porous surface or a substance in which at least one of
silica gel and cellulose is mixed is suitable.
[0037] In this manner, the chip for analysis of a target substance
10 shown in FIG. 1 can be obtained.
[0038] Next, the first target substance analysis method of the
present invention can be performed using the first chip for
analysis of a target substance of the present invention. The first
target substance analysis method is characterized, for example, by
using the first chip for analysis of a target substance of the
present invention and including the following steps (a1) to
(d1):
(a1) a step of forming a shutter portion for blocking the flow
channel by supplying pressure from the pressure supply port to
raise a site above the shutter-forming non-bonded area; (b1) a step
of forming the flow channel and extraction chamber by supplying
pressure from the through-hole to raise a site above the flow
channel-forming non-bonded area and a site above the extraction
chamber-forming non-bonded area; (c1) a step of injecting an
analysis sample into the flow channel and the extraction chamber;
and (d1) a step of capturing the target substance in the analysis
sample that is bound to the magnetic particle by generating a
magnetic field at at least one of the undersurface of the third
substrate directly below the end of the extraction chamber at the
opposite side of the through-hole and the top surface of the first
flexible substrate directly above the end of the extraction chamber
at the opposite side of the through-hole.
[0039] In the first target substance analysis method of the present
invention, there is no particular limitation on the order of the
above steps. For example, the steps may be performed in the order
from step (a1) to step (d1). The shutter portion formation step
(a1) and the flow channel and extraction chamber formation step
(b1) may be performed simultaneously or either of the steps (a1)
and (b1) may be performed in advance, for example. Furthermore, the
analysis sample injection step (c1) may be performed simultaneously
with the flow channel and extraction chamber formation step (b1),
for example.
[0040] As the first target substance analysis method of the present
invention, an example of the usage of the chip for analysis of a
target substance 10 shown in FIG. 1 will be described with
reference to FIG. 2. The aspect shown in FIG. 2 is an example and
the present invention is not limited to this aspect.
[0041] First, as shown in FIG. 2(A), an adapter 14 is provided at
the opening portion of the through-hole 7 serving as an
introduction portion of liquid or gas, and an injection tube 15 is
connected to the adapter 14. The shape of the adapter 14 is not
limited to that shown in FIG. 2(A). For example, the adapter 14 may
not be in the form of partially inserted in the through-hole 7 but
may be in the form of directly fixed to the top surface of the
first flexible substrate 1. Furthermore, the injection tube 15 may
be directly connected to the through-hole 7 without using the
adapter 14. As the material for the adapter 14, although a silicone
rubber such as PDMS is preferable, any other material can be used.
In the case where a material other than PDMS is used, an
appropriate adhesive agent may be used for fixing the adapter 14 to
the top surface of the first flexible substrate 1. An example of
the injection tube 15 includes a Teflon (registered trademark)
tube. One end of the injection tube 15 is fixed to the adapter 14
using an appropriate adhesive agent. The other end of the injection
tube 15 is connected to an appropriate undiluted solution supply
means, an appropriate pressure application means (for example, a
micro-pump, a syringe, or the like), and the like although it is
not shown.
[0042] The adapter 14 to which the injection tube 15 is connected
is provided also at each of the pressure supply ports 18a and 18b
although it is not shown. Then, gas is injected at high pressure
from the injection tube 15 via the pressure supply port 18b.
Thereby, as shown in FIG. 2(B), a site above the shutter-forming
non-bonded area 12b is raised to form the shutter-forming void 17b.
Specifically, only a part of the first flexible substrate 1 and a
part of the second flexible substrate 2 positioned above the
shutter-forming non-bonded area 12b are raised from the top surface
of the third substrate 3 to form the shutter-forming void 17b. The
shutter-forming void 17b formed by raising is also referred to as a
shutter portion (hereinafter, the same applies). The gas is, for
example, air or the like, and the level of high pressure is, for
example, in the range from 10 kPa to 300 kPa (hereinafter, the same
applies).
[0043] Next, a liquid analysis sample to be analyzed is injected
into the chip for analysis of a target substance 10. In the present
invention, there is no particular limitation on the type of the
analysis sample, and, for example, the type of the analysis sample
can be selected appropriately according to the type of the target
substance. Examples of the target substance include cells and
intracellular components, and specific examples thereof include
nucleic acids such as DNA and RNA. In the case where the target
substance is the intracellular component such as the nucleic acids,
the analysis sample may be, for example, a sample in which a target
substance is eluted from a cell, i.e., an elution sample of a cell
(also referred to as a target substance-eluted sample), or a sample
in which a target substance is not eluted from a cell, i.e., a
sample that contains a cell. In the latter case, for example, the
target substance such as the nucleic acid or the like may be eluted
from a cell in the analysis sample in the chip for analysis of a
target substance 10.
[0044] Specifically, gas is injected at high pressure from the
injection tube 15 after injecting the analysis sample into the
through-hole 7 or the analysis sample is injected into the
through-hole 7 with application of positive pressure. Thereby, as
shown in FIG. 2(B), a site above the flow channel-forming
non-bonded area 11 and a site above the extraction chamber-forming
non-bonded area 5 are raised and the flow channel 8 and the
extraction chamber 6 are formed. Specifically, only parts of the
first flexible substrate 1 positioned above the flow
channel-forming non-bonded area 11 and the extraction
chamber-forming non-bonded area 5 are raised from the top surface
of the second flexible substrate 2 to form the flow channel 8 and
extraction chamber 6. On this occasion, a site above the flow
channel-forming non-bonded area 11 positioned further ahead of the
shutter-forming void 17b, i.e., a site above the flow
channel-forming non-bonded area 11 positioned at the downstream
side of the shutter-forming void 17b is blocked by the
shutter-forming void 17b, and therefore the flow channel is not
formed. At this time, in the extraction chamber 6, the target
substance contained in the analysis sample that has been injected
binds to the magnetic particle 16.
[0045] In the case where the analysis sample is the sample that
contains a cell as described above, for example, an elution reagent
that causes a target substance such as a nucleic acid to be eluted
from the cell may be injected into the chip for analysis of a
target substance 10 before, at the same time as, or after the
injection of the analysis sample. The method of injection is, for
example, the same as that described for the analysis sample. Then,
the target substance eluted from the cell by the elution reagent
binds to the magnetic particle 16 in the extraction chamber 6. It
is also possible to preliminarily place the elution reagent, for
example, at the extraction chamber-forming non-bonded area 5 or at
the flow channel-forming non-bonded area 11 between the
through-hole 7 and the extraction chamber-forming non-bonded area
5.
[0046] Next, a washing reagent is injected into the chip for
analysis of a target substance 10. There is no particular
limitation on the method of injection of the washing reagent, and,
for example, the washing reagent is injected from the injection
tube 15 via the through-hole 7 in the same manner as the analysis
sample.
[0047] Thereafter, a magnetic field is generated at the
undersurface of the third substrate 3. Specifically, a magnetic
field is generated at the undersurface of the third substrate 3
directly below the end of the extraction chamber 6 at the opposite
side of the through-hole 7. Thereby, in the extraction chamber 6, a
target substance such as DNA that is bound to the magnetic particle
16 is captured. In this manner, by generating a magnetic field at
the undersurface of the third substrate 3, the leak of the magnetic
particle 16 to the flow channel 8 further ahead of the extraction
chamber 6 can be prevented even in the case where the flow channel
8 is formed at the downstream side of the extraction chamber 6. The
magnetic field may be generated, for example, at the top surface
side of the first flexible substrate 1. Specifically, the magnetic
field may be generated at the top surface side of the first
flexible substrate 1 directly above the end of the extraction
chamber 6 at the opposite side of the through-hole 7.
[0048] There is no particular limitation on the method of
generating a magnetic field, and an example thereof includes a
method of making the chip for analysis of a target substance 10
into contact with a magnet 13 such as an electromagnet or a
permanent magnet such as an alnico magnet, a ferrite magnet, a
neodymium magnet, or a samarium-cobalt magnet.
[0049] Next, the pressure of the gas injecting from the
through-hole 7 and the pressure supply port 18b is set about
atmospheric pressure. Thereby, as shown in FIG. 2(C), the
shutter-forming void 17b and the voids of the flow channel 8 and
the extraction chamber 6 are vanished. Thereafter, gas is injected
at high pressure from the injection tube 15 via the through-hole 7.
Thereby, substances excluding the target substance that is bound to
the magnetic particle 16, e.g., the washing reagent and the like
can be discharged from the flow channel 8. In this manner, the chip
for analysis of a target substance of this Embodiment allows
extraction of a target substance such as DNA from the analysis
sample efficiently by the magnetic particle 16. Since extraction of
the target substance can be also referred to as separation of the
target substance from the analysis sample, the extraction chamber
can be also referred to as, for example, a separation chamber of
the target substance.
Embodiment 2
[0050] FIG. 3 shows an example of the configuration of the second
chip for analysis of a target substance of the present invention.
The aspect shown in FIG. 3 is an example and the present invention
is not limited to this aspect. In FIG. 3, identical parts to those
shown in FIGS. 1 and 2 are indicated with identical numerals and
symbols. The chip for analysis of a target substance 10 shown in
FIG. 3 has the configuration identical to that of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2 except
that the extraction chamber-forming non-bonded area 5 has the
function of a mixing chamber-forming non-bonded area 9 and does not
contain the magnetic particle 16.
[0051] In the analysis of the target substance, for example,
various reagents are used. The chip for analysis of a target
substance of this Embodiment allows mixing of the reagent and the
analysis sample or the target substance in the analysis sample in
the mixing chamber 19 in the manner described below. There is no
particular limitation on the reagent, and can be selected
appropriately according to, for example, the type of the analysis
sample, the type of the target substance, and the analysis method.
Specific examples of the reagent include the aforementioned elution
reagent that causes a target substance to be eluted from the cell,
a reaction reagent that reacts with the target substance, and the
washing reagent.
[0052] Next, the second target substance analysis method of the
present invention can be performed using the second chip for
analysis of a target substance of the present invention. The second
target substance analysis method is characterized, for example, by
using the second chip for analysis of a target substance of the
present invention and including the following steps (a2) to
(f2):
(a2) a step of forming a shutter portion for blocking the flow
channel by supplying pressure from the pressure supply port to
raise a site above the shutter-forming non-bonded area at the far
side of the through-hole relative to the mixing chamber-forming
non-bonded area; (b2) a step of forming the flow channel and the
mixing chamber by supplying pressure from the through-hole to raise
a site above the flow channel-forming non-bonded area and a site
above the mixing chamber-forming non-bonded area; (c2) a step of
injecting an analysis sample into the flow channel and the mixing
chamber; (d2) a step of injecting a reagent into the flow channel
and the mixing chamber; (e2) a step of forming a shutter portion
for blocking the flow channel by supplying pressure from the
pressure supply port to raise a site above the shutter-forming
non-bonded area at the near side of the through-hole relative to
the mixing chamber-forming non-bonded area; and (f2) a step of
mixing the target substance in the analysis sample and the reagent
in the mixing chamber by applying pressure to the top surface of
the first flexible substrate above the mixing chamber to deform the
mixing chamber.
[0053] In the second target substance analysis method of the
present invention, there is no particular limitation on the order
of the above steps. For example, the steps may be performed in the
order from step (a2) to step (f2). The shutter portion formation
step (a2) and the flow channel and mixing chamber formation step
(b2) may be performed simultaneously or either of the steps (a2)
and (b2) may be performed in advance, for example. The analysis
sample injection step (c2) and the reagent injection step (d2) may
be performed simultaneously or either of the steps (c2) and (d2)
may be performed in advance, for example. Furthermore, the analysis
sample injection step (c2) and the reagent injection step (d2) may
be performed simultaneously with the flow channel and mixing
chamber formation step (b2), for example.
[0054] As the second target substance analysis method of the
present invention, an example of the usage of the chip for analysis
of a target substance 10 will be described with reference to FIG.
3. First, as shown in FIGS. 3(A) and 3(B), the steps to the
analysis sample injection step (steps before washing reagent
injection) are performed in the same manner as in Embodiment 1. At
this time, the extraction chamber 6 is formed in Embodiment 1
whereas the mixing chamber 19 is formed in this Embodiment.
[0055] Note here that the aforementioned various reagents may be
injected into the chip for analysis of a target substance 10, for
example, before, at the same time as, or after the injection of the
analysis sample. The method of injection is, for example, the same
as that described for the analysis sample. In the case where the
analysis sample is the sample that contains a cell as described
above, for example, the elution reagent, the reaction reagent that
reacts with the eluted target substance, the washing reagent that
washes the target substance, and the like may be injected as the
reagent. Furthermore, in the case where the analysis sample is the
target substance-eluted sample as described above, for example, the
reaction reagent, the washing reagent, and the like may be injected
as the reagent. It is also possible to preliminarily place the
elution reagent and the reaction reagent, for example, at the
mixing chamber-forming non-bonded area 9 or at the flow
channel-forming non-bonded area 11 between the through-hole 7 and
the mixing chamber-forming non-bonded area 9.
[0056] Next, gas is injected at high pressure from the injection
tube 15 via the pressure supply port 18a. Thereby, as shown in FIG.
3(C), a site above the shutter-forming non-bonded area 12a is
raised to form the shutter-forming void 17a. Specifically, only a
part of the first flexible substrate 1 and a part of the second
flexible substrate 2 positioned above the shutter-forming
non-bonded area 12a are raised from the top surface of the third
substrate 3 to form the shutter-forming void 17a. The gas is, for
example, air or the like, and the level of high pressure is, for
example, in the range from 10 kPa to 300 kPa (hereinafter, the same
applies). Next, the pressure of the gas injecting from the
through-hole 7 is set about atmospheric pressure. Thereby, as shown
in FIG. 3(C), the void of the flow channel 8 at the upstream side
of the shutter-forming void 17a is vanished.
[0057] Next, as shown in FIG. 3(D), the mixing chamber 19 is
deformed by applying pressure to the top surface of the first
flexible substrate 1 above the mixing chamber 19. Thereby, the
target substance and the reagent are mixed in the mixing chamber
19. There is no particular limitation on the method of applying
pressure to the position above the mixing chamber 19, and, for
example, gas may be sprayed at high pressure or an object may be
pressed.
[0058] Next, the pressure of the gas injecting from the pressure
supply ports 18a and 18b is set about atmospheric pressure.
Thereby, as shown in FIG. 3(E), the shutter-forming voids 17a and
17b and the void of the mixing chamber 19 are vanished. Thereafter,
gas is injected at high pressure from the injection tube 15 via the
through-hole 7. Thereby, the target substance mixed with the
reagent can be forwarded to the next step.
[0059] In the chip for analysis of a target substance of this
Embodiment, the same magnetic particle as described in Embodiment 1
may be placed at the mixing chamber-forming non-bonded area 9. In
this case, the mixing chamber 19 also has the function of an
extraction chamber.
Embodiment 3
[0060] In the third chip for analysis of a target substance of the
present invention, as described above, two shutter-forming
non-bonded areas are formed on the bonding surface of the second
flexible substrate and the third substrate at the near side of the
through-hole relative to the first mixing chamber-forming
non-bonded area and at the far side of the through-hole relative to
the second mixing chamber-forming non-bonded area. In the third
chip for analysis of a target substance, for example, the third
shutter-forming non-bonded area may be further formed on the
bonding surface of the second flexible substrate and the third
substrate in a band-like manner such that the third shutter-forming
non-bonded area and the flow channel-forming non-bonded area
intersect above and below via the second flexible substrate being
interposed therebetween. This shutter-forming non-bonded area may
be formed at the far side of the through-hole relative to the first
mixing chamber-forming non-bonded area, for example. In this case,
the flow channel at the upstream side and the downstream side of
the first mixing chamber can be blocked respectively by the shutter
portions.
[0061] FIG. 4 shows an example of the configuration of the third
chip for analysis of a target substance of the present invention.
The aspect shown in FIG. 4 is an example and the present invention
is not limited to this aspect. In FIG. 4, identical parts to those
shown in FIGS. 1 to 3 are indicated with identical numerals and
symbols. The chip for analysis of a target substance 10 shown in
FIG. 4 has the configuration identical to that of the chip for
analysis of a target substance shown in FIG. 3 except that the chip
for analysis of a target substance 10 shown in FIG. 4 includes two
mixing chamber-forming non-bonded areas (9a and 9b), four
shutter-forming non-bonded areas (12a to 12d), and four pressure
supply ports. Although it is not shown, four pressure supply ports
are referred to as pressure supply ports 18a to 18d for convenience
sake.
[0062] Although it is not shown, the shutter-forming non-bonded
areas 12c and 12d are respectively in contact with the pressure
supply ports 18c and 18d that come through the first flexible
substrate 1 and the second flexible substrate 2 as in the case of
the shutter-forming non-bonded areas 12a and 12b shown in FIG.
1(A). The pressure supply ports 18c and 18d may be formed on the
third substrate 3 in such a manner that they come through the third
substrate 3 so as to be in contact with the shutter-forming
non-bonded areas 12c and 12d. In this Embodiment, the
shutter-forming non-bonded areas 12b and 12c and the pressure
supply ports 18b and 18c are optional components and are not
indispensable, although the chip for analysis of a target substance
of this Embodiment preferably includes these components.
Furthermore, in this Embodiment, the shutter-forming non-bonded
areas 12b and 12c and the pressure supply ports 18b and 18c may be
respectively formed as one component, and the number of the
non-bonded areas and the pressure supply ports may be respectively
three.
[0063] Next, the third target substance analysis method of the
present invention can be performed using the third chip for
analysis of a target substance of the present invention. The third
target substance analysis method is characterized, for example, by
using the third chip for analysis of a target substance of the
present invention and including the following steps (a3) to
(f3):
(a3) a step of forming the flow channel, the first mixing chamber,
and the second mixing chamber by supplying pressure from the
through-hole to raise a site above the flow channel-forming
non-bonded area, a site above the first mixing chamber-forming
non-bonded area, and a site above the second mixing chamber-forming
non-bonded area; (b3) a step of injecting an analysis sample into
the flow channel and the mixing chamber; (c3) a step of injecting a
reagent into the flow channel and the mixing chamber; (d3) a step
of forming a shutter portion for blocking the flow channel by
supplying pressure from the pressure supply port to raise a site
above the shutter-forming non-bonded area at the near side of the
through-hole relative to the first mixing chamber-forming
non-bonded area; (e3) a step of forming a shutter portion for
blocking the flow channel by supplying pressure from the pressure
supply port to raise a site above the shutter-forming non-bonded
area at the far side of the through-hole relative to the second
mixing chamber-forming non-bonded area; and (f3) a step of mixing
the target substance in the analysis sample and the reagent by
moving them between the first mixing chamber and the second mixing
chamber.
[0064] In the third target substance analysis method of the present
invention, there is no particular limitation on the order of the
above steps. For example, the steps may be performed in the order
from step (a3) to step (f3). The step (d3) may be performed, for
example, before, at the same time as, or after the step (a3) and is
preferably performed before the steps (b3) and (c3). The step (e3)
is preferably performed, for example, after the steps (b3) and
(c3).
[0065] Furthermore, in the present invention, the formation of the
first mixing chamber and the second mixing chamber in the step (a3)
may be performed, for example, as a separated step. In this case,
the step (a3) may be the steps (a3-1) and (a3-2) described
below:
(a3-1) a step of forming the flow channel and the first mixing
chamber by supplying pressure from the through-hole to raise a site
above the flow channel-forming non-bonded area between the
through-hole and the first mixing chamber-forming non-bonded area
and a site above the first mixing chamber-forming non-bonded area;
and (a3-2) a step of forming the flow channel and the second mixing
chamber by applying pressure to a site above the first mixing
chamber to deform the first mixing chamber so as to raise a site
above the flow channel-forming non-bonded area between the first
mixing chamber-forming non-bonded area and the second mixing
chamber-forming non-bonded and a site above the second mixing
chamber-forming non-bonded area.
[0066] The first mixing chamber formation step (a3-1) is preferably
performed before or at the same time as the analysis sample
injection step (b3) and the reagent injection step (c3). The second
mixing chamber formation step (a3-2) may be performed, for example,
before, after, or during the shutter portion formation steps (d3)
and (e3).
[0067] Furthermore, in the case where the third chip for analysis
of a target substance includes the third shutter-forming non-bonded
area, in advance of the first mixing chamber formation step (a3-1)
or in advance of the analysis sample injection step (b3) and the
reagent injection step (c3), a shutter portion may be formed by
raising a site above the third shutter-forming non-bonded area.
[0068] The chip for analysis of a target substance 10 shown in FIG.
4 is used, for example, as follows. First, as shown in FIG. 4(A),
in the same manner as in Embodiment 1, the adapter 14 to which the
injection tube 15 is connected is provided at each of the
through-hole 7 and the pressure supply ports 18a to 18d.
[0069] Next, gas is injected at high pressure from the injection
tube 15 via the pressure supply port 18b. Thereby, as shown in FIG.
4(B), a site above the shutter-forming non-bonded area 12b is
raised to form the shutter-forming void 17b. Specifically, only a
part of the first flexible substrate 1 and a part of the second
flexible substrate 2 positioned above the shutter-forming
non-bonded area 12b are raised from the top surface of the third
substrate 3 to form the shutter-forming void 17b. The gas is, for
example, air or the like, and the level of high pressure is, for
example, in the range from 10 kPa to 300 kPa (hereinafter, the same
applies).
[0070] Next, gas is injected at high pressure from the injection
tube 15 after injecting the analysis sample into the through-hole 7
or the analysis sample is injected into the through-hole 7 with
application of positive pressure. Thereby, as shown in FIG. 4(B), a
site above the flow channel-forming non-bonded area 11 and a site
above the first mixing chamber-forming non-bonded area 9a are
raised and the flow channel 8 and the first mixing chamber 19a are
formed. Specifically, only parts of the first flexible substrate 1
positioned above the flow channel-forming non-bonded area 11 and
the first mixing chamber-forming non-bonded area 9a are raised from
the top surface of the second flexible substrate 2 to form the flow
channel 8 and the first mixing chamber 19a. On this occasion, a
site above the flow channel-forming non-bonded area 11 positioned
further ahead of the shutter-forming void 17b, i.e., a site above
the flow channel-forming non-bonded area 11 positioned at the
downstream side of the shutter-forming void 17b is blocked by the
shutter-forming void 17b, and therefore the flow channel is not
formed. At this time, in the same manner as in Examples 1 and 2,
for example, the reagent is injected into the chip for analysis of
a target substance 10. In the case where the analysis sample is the
sample that contains a cell as described above and the elution
reagent is used as the reagent, a target substance such as DNA is
eluted from the cell in the first mixing chamber 19a.
[0071] Next, gas is injected at high pressure from the injection
tube 15 via the pressure supply port 18a. Thereby, as shown in FIG.
4(C), a site above the shutter-forming non-bonded area 12a is
raised to form the shutter-forming void 17a. Specifically, only a
part of the first flexible substrate 1 and a part of the second
flexible substrate 2 positioned above the shutter-forming
non-bonded area 12a are raised from the top surface of the third
substrate 3 to form the shutter-forming void 17a. The gas is, for
example, air or the like, and the level of high pressure is, for
example, in the range from 10 kPa to 300 kPa (hereinafter, the same
applies). Next, the pressure of the gas injecting from the
through-hole 7 is set about atmospheric pressure. Thereby, as shown
in FIG. 4(C), the void of the flow channel 8 at the upstream side
of the shutter-forming void 17a is vanished.
[0072] Next, the pressure of the gas injecting from the pressure
supply port 18b is set about atmospheric pressure. Thereby, as
shown in FIG. 4(D), the void of the shutter-forming void 17b is
vanished. Thereafter, gas is injected at high pressure from the
injection tube 15 via the pressure supply port 18d. Thereby, as
shown in FIG. 4(D), a site above the shutter-forming non-bonded
area 12d is raised to form the shutter-forming void 17d.
Specifically, only a part of the first flexible substrate 1 and a
part of the second flexible substrate 2 positioned above the
shutter-forming non-bonded area 12d are raised from the top surface
of the third substrate 3 to form the shutter-forming void 17d.
[0073] Then, pressure is applied to the top surface of the first
flexible substrate 1 above the first mixing chamber 19a. Thereby, a
part of the first flexible substrate 1 positioned above the flow
channel-forming non-bonded area 11 between the first mixing
chamber-forming non-bonded area 9a and the second mixing
chamber-forming non-bonded area 9b and a part of the first flexible
substrate 1 positioned above the second mixing chamber-forming
non-bonded area 9b are raised from the top surface of the second
flexible substrate 2 to form the flow channel 8 and the second
mixing chamber 19b. Thereby, the target substance and the reagent
are moved from the first mixing chamber 19a to the second mixing
chamber 19b.
[0074] Next, as shown in FIG. 4(E), pressure is applied to the top
surface of the first flexible substrate 1 above the second mixing
chamber 19b. Thereby, the target substance and the reagent are
moved from the second mixing chamber 19b to the first mixing
chamber 19a.
[0075] Thereafter, pressure is alternately applied to the top
surface of the first flexible substrate 1 positioned above the
first mixing chamber 19a and the top surface of the first flexible
substrate 1 positioned above the second mixing chamber 19b to
alternately deform the first mixing chamber 19a and the second
mixing chamber 19b. Thereby, the target substance and the reagent
are mixed by moving between the first mixing chamber 19a and the
second mixing chamber 19b. There is no particular limitation on the
method of applying pressure to a site above the first mixing
chamber 19a and a site above the second mixing chamber 19b, and,
for example, gas may be sprayed at high pressure or an object may
be pressed.
[0076] Furthermore, the method of mixing the target substance and
the reagent by moving them between the first mixing chamber 19a and
the second mixing chamber 19b is not limited to the method of
alternately applying pressure to a site above the first mixing
chamber 19a and a site above the second mixing chamber 19b to
alternately deform the first mixing chamber 19a and the second
mixing chamber 19b, and any method can be employed. For example,
the target substance and the reagent may be mixed by applying air
pressure between the first mixing chamber 19a and the second mixing
chamber 19b to move the target substance and the reagent between
the first mixing chamber 19a and the second mixing chamber 19b.
[0077] Next, the pressure of the gas injecting from the
through-hole 7 and the pressure supply ports 18a and 18d is set
about atmospheric pressure. Thereby, as shown in FIG. 4(F), the
shutter-forming voids 17a and 17d and the voids of the flow channel
8, the first mixing chamber 19a, and the second mixing chamber 19b
are vanished. Thereafter, gas is injected at high pressure from the
injection tube 15 via the through-hole 7. Thereby, the target
substance mixed with the reagent can be forwarded to the next
step.
[0078] In the chip for analysis of a target substance of this
Embodiment, the same magnetic particle as described in Embodiment 1
may be placed at at least one of the first mixing chamber-forming
non-contact (non-bonded) area 9a and the second mixing
chamber-forming non-bonded area 9b. In this case, at least one of
the first mixing chamber 19a and the second mixing chamber 19b has
the function of an extraction chamber.
Embodiment 4
[0079] FIG. 5 shows another example of the configuration of the
chip for analysis of a target substance of the present invention.
The aspect shown in FIG. 5 is an example and the present invention
is not limited to this aspect. In FIG. 5, identical parts to those
shown in FIGS. 1 and 2 are indicated with identical numerals and
symbols. A chip for analysis of a target substance 20 shown in FIG.
5 includes, in addition to the configuration of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2, a washing
reagent supply portion 30, a PCR reaction reagent supply portion
40, a washing reagent recovery portion 70, a PCR amplification
portion 50, a shutter-forming non-bonded area 12m, a pressure
supply port 18m, and an electrophoresis analysis portion 60 as main
components. The washing reagent supply portion 30, the PCR reaction
reagent supply portion 40, the washing reagent recovery portion 70,
the PCR amplification portion 50, and the electrophoresis analysis
portion 60 each include a laminate in which the first flexible
substrate 1, the second flexible substrate 2, and the third
substrate 3 are laminated. The material of each of the first
flexible substrate 1, the second flexible substrate 2, and the
third substrate 3 is the same as that of the chip for analysis of a
target substance 10 shown in FIGS. 1 and 2. The shutter-forming
non-bonded area 12m and the pressure supply port 18m can be formed
in the same manner as the corresponding components of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2.
[0080] The washing reagent supply portion 30 includes a
through-hole 37, a flow channel-forming non-bonded area 31, a
shutter-forming non-bonded area 12e, and a pressure supply port 18e
as main components. These components can be formed in the same
manner as the corresponding components of the chip for analysis of
a target substance 10 shown in FIGS. 1 and 2. The flow
channel-forming non-bonded area 31 is in contact with the
extraction chamber-forming non-bonded area 5 of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2. In the
chip for analysis of a target substance 20 of this Embodiment, the
washing reagent supply portion 30 is an optional component and is
not indispensable, although the chip for analysis of a target
substance 20 preferably includes the washing reagent supply portion
30. In the case where the chip for analysis of a target substance
20 does not include the washing reagent supply portion 30, a
washing reagent may be supplied from the through-hole 7 of the chip
for analysis of a target substance 10 shown in FIGS. 1 and 2.
[0081] The PCR reaction reagent supply portion 40 includes a
through-hole 47, a flow channel-forming non-bonded area 41, a
shutter-forming non-bonded area 12f, and a pressure supply port 18f
as main components. These components can be formed in the same
manner as the corresponding components of the chip for analysis of
a target substance 10 shown in FIGS. 1 and 2. The flow
channel-forming non-bonded area 41 is in contact with the
extraction chamber-forming non-bonded area 5 of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2. In the
chip for analysis of a target substance 20 of this Embodiment, the
PCR reagent supply portion 40 is an optional component and is not
indispensable, although the chip for analysis of a target substance
20 preferably includes the PCR reagent supply portion 40. In the
case where the chip for analysis of a target substance 20 does not
include the PCR reagent supply portion 40, a PCR reagent may be
supplied from the through-hole 7 of the chip for analysis of a
target substance 10 shown in FIGS. 1 and 2.
[0082] The washing reagent recovery portion 70 includes a flow
channel-forming non-bonded area 71, shutter-forming non-bonded
areas 12n and 12o, pressure supply ports 18n and 18o, and a waste
tank 78 as main components. The components except for the waste
tank 78 can be formed in the same manner as the corresponding
components of the chip for analysis of a target substance 10 shown
in FIGS. 1 and 2. The waste tank 78 can be formed in the same
manner as the extraction chamber-forming non-bonded area 5 of the
chip for analysis of a target substance 10 shown in FIGS. 1 and
2.
[0083] In the PCR amplification portion 50, the flow
channel-forming non-bonded area 11 led out from the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2 is split
into eight flow channel-forming non-bonded areas 51a to 51h via the
shutter-forming non-bonded areas 12g to 12l and pressure supply
ports 18g to 18l. The number of the split of the flow
channel-forming non-bonded area is not limited to eight and can be
increased or decreased appropriately according to a desired
analysis accuracy of the target substance. The shutter-forming
non-bonded areas 12g to 12l, the pressure supply ports 18g to 18l,
and the flow channel-forming non-bonded areas 51a to 51h can be
formed in the same manner as the corresponding components of the
chip for analysis of a target substance 10 shown in FIGS. 1 and 2.
Eight flow channel-forming non-bonded areas 51a to 51h are
respectively in contact with eight reaction tanks 52a to 52h. On
the flow channel-forming non-bonded areas 51a to 51h, in the
vicinity of contact points with the reaction tanks 52a to 52h,
shutter-forming non-bonded areas 12p to 12z and 12a to 12e and
pressure supply ports 18p to 18z and 18a to 18e are respectively
formed. There is no particular limitation on the method of forming
the reaction tanks 52a to 52h, and, for example, a formation method
in a conventionally known PCR chip can be employed. The
shutter-forming non-bonded areas 12p to 12z and 12a to 12e and the
pressure supply ports 18p to 18z and 18a to 18e can be formed in
the same manner as the corresponding components of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2.
[0084] Although it is not shown, at at least one of positions of
the undersurface of the third substrate 3 directly below the
reaction tanks 52a to 52h and positions of the top surface of the
first flexible substrate 1 directly above the reaction tanks 52a to
52h, heating means such as heaters are placed.
[0085] The electrophoresis analysis portion 60 includes reagent
tanks 67a to 67h, through-holes 68a to 68h, flow channel-forming
non-bonded areas 61a to 61h and 62a to 62h, waste tanks 65a to 65h
and 66a to 66h, and electrodes 67i to 67p, 68i to 68p, 65i to 65p,
and 66i to 66p. The reagent tanks 67a to 67h are formed so as to be
in contact with the flow channel-forming non-bonded areas 51a to
51h of the PCR amplification portion 50 via the shutter-forming
non-bonded area 12m and the pressure supply port 18m. The flow
channel-forming non-bonded areas 61a to 61h are formed so as to be
in contact with the reagent tanks 67a to 67h at one end and be in
contact with the waste tanks 65a to 65h at the other end. The flow
channel-forming non-bonded areas 62a to 62h are formed so as to
intersect with the flow channel-forming non-bonded areas 61a to
61h, be in contact with the through-holes 68a to 68h at one end,
and be in contact with the waste tanks 66a to 66h at the other end.
At the reagent tanks 67a to 67h, the through-holes 68a to 68h, and
the waste tanks 65a to 65h and 66a to 66h, the electrodes 67i to
67p, 68i to 68p, 65i to 65p, and 66i to 66p are respectively
placed. It is possible to apply voltages to the electrodes 67i to
67p, 68i to 68p, 65i to 65p, and 66i to 66p from above the first
flexible substrate 1 or below third substrate 3. The through-holes
67a to 67h and the flow channel-forming non-bonded areas 61a to 61h
and 62a to 62h can be formed in the same manner as the
corresponding components of the chip for analysis of a target
substance 10 shown in FIGS. 1 and 2. Note here that, instead of the
flow channel-forming non-bonded areas 61a to 61h and 62a to 62h,
grooves formed on the third substrate 3 according to a
conventionally known method may be used as flow channels. The
groove has, for example, a width of about 100 .mu.m and a depth of
about 30 .mu.m. The reagent tanks 67a to 67h and the waste tanks
65a to 65h and 66a to 66h can be formed in the same manner as the
extraction chamber-forming non-bonded area 5 of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2. As the
electrodes 67i to 67p, 68i to 68p, 65i to 65p, and 66i to 66p,
conventionally known ones can be used.
[0086] Although it is not shown, at at least one of positions of
the undersurface of the third substrate 3 below the flow
channel-forming non-bonded areas 62a to 62h and positions of the
top surface of the first flexible substrate 1 above the flow
channel-forming non-bonded areas 62a to 62h, optical analysis means
such as absorbance measuring apparatuses are placed.
[0087] The chip for analysis of a target substance 20 of this
Embodiment may include the configuration of the chip for analysis
of a target substance 10 including the mixing chamber-forming
non-bonded area shown in FIG. 3 or FIG. 4 instead of the chip for
analysis of a target substance 10 including the extraction
chamber-forming non-bonded area 5 shown in FIGS. 1 and 2.
Furthermore, the chip for analysis of a target substance 20 of this
Embodiment may further include a through-hole and a flow
channel-forming non-bonded area for dry air supply that are formed
so as to be in contact with the extraction chamber-forming
non-bonded area or the mixing chamber-forming non-bonded area.
[0088] The size of the chip for analysis of a target substance 20
of this Embodiment is as follows. That is, for example, the length
is in the range from 50 mm to 300 mm and the width is in the range
from 20 mm to 100 mm. Since the chip for analysis of a target
substance of the present invention is compact as described above,
it allows a small installation space.
[0089] Furthermore, the thickness of the chip for analysis of a
target substance 20 of this Embodiment excluding the mechanism for
generating a magnetic field in the configuration of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2, the
heating means in the PCR amplification portion 50, and the optical
analysis means in the electrophoresis analysis portion 60 is, for
example, in the range from 0.5 mm to 5 mm. Therefore, the chip for
analysis of a target substance 20 of this Embodiment can be carried
around without fixing at a predetermined space.
[0090] The chip for analysis of a target substance 20 shown in FIG.
5 is used, for example, as follows. First, in the same manner as in
Embodiment 1, the target substance such as DNA is extracted from
the analysis sample using the configuration of the chip for
analysis of a target substance 10 shown in FIGS. 1 and 2 and the
washing reagent supply portion 30. The time required for extracting
the target substance is, for example, about 5 minutes.
[0091] Next, the target substance that is bound to a magnetic
particle is transferred to the washing reagent recovery portion 70
by supplying the PCR reaction reagent from the PCR reaction reagent
supply portion 40. Next, in the washing reagent recovery portion
70, a solution obtained by removing the washing reagent from a
mixture of the target substance, the washing reagent, the PCR
reaction reagent, and the like is transferred to the PCR
amplification portion 50. Then, PCR amplification is performed by a
conventionally known method such as a method of applying a
temperature cycle to the target substance and the PCR reaction
reagent stored in the reaction tanks 52a to 52h. The time required
for this PCR amplification is, for example, in the range from 10
minutes to 60 minutes and preferably about 15 minutes.
[0092] Next, after PCR amplification, potential differences are
generated between the reagent tanks 67a to 67h and the waste tanks
65a to 65h respectively by transferring the amplification products
of the target substance to the reagent tanks 67a to 67h of the
electrophoresis analysis portion 60 and applying voltages to the
electrodes 67i to 67p and 65i to 65p. Thereby, the flow channels
formed above the flow channel-forming non-bonded areas 61a to 61h
are filled with the amplification products of the target substance.
Next, potential differences are generated between the through-holes
68a to 68h and the waste tanks 66a to 66h respectively by supplying
an electrophoresis solution from the through-holes 68a to 68h and
applying voltages to the electrodes 68i to 68p and 66i to 66p.
Thereby, electrophoresis analysis is performed by introducing a
small amount of amplification products of the target substance from
the intersection site of the flow channel-forming non-bonded areas
61a to 61h and 62a to 62h to the flow channels formed above the
flow channel-forming non-bonded areas 62a to 62h. The time required
for this electrophoresis analysis is, for example, about 5 minutes.
Such an electrophoresis analysis method is conventionally
known.
[0093] In this manner, the chip for analysis of a target substance
20 of this Embodiment allows extraction, amplification, and
analysis of a target substance such as DNA with less effort and
less time such as from about 20 minutes to about 70 minutes.
[0094] The chip for analysis of a target substance 20 shown in FIG.
5 includes the PCR amplification portion 50 and the electrophoresis
analysis portion 60. However, this Embodiment is not limited
thereto. The chip for analysis of a target substance of this
Embodiment may be the one that performs electrophoresis analysis
without performing PCR amplification. Furthermore, the chip for
analysis of a target substance of this Embodiment may be the one
that analyzes the target substance by a method other than
electrophoresis analysis such as chemiluminescence, fluorescence,
or enzyme coloration. For example, analysis of the target substance
such as DNA may be performed by a conventional known method such as
an intercalation method or a method using a fluorescent-labeled
probe.
[0095] The invention of the present application was described above
with reference to the embodiments. However, the invention of the
present application is not limited to the above-described
embodiments. Various changes that can be understood by those
skilled in the art can be made in the configurations and details of
the invention of the present application within the scope of the
invention of the present application.
[0096] This application claims priority from Japanese Patent
Application No. 2012-063645 filed on Mar. 21, 2012. The entire
subject matter of the Japanese Patent Application is incorporated
herein by reference.
INDUSTRIAL APPLICABILITY
[0097] As described above, the chip for analysis of a target
substance of the present invention is compact and allows analysis
of a target substance such as DNA with less time and effort. The
chip for analysis of a target substance of the present invention
can be applied to a wide range of uses including, for example, DNA
analysis in a criminal investigation.
EXPLANATION OF REFERENCE NUMERALS
[0098] 1 first flexible substrate [0099] 2 second flexible
substrate [0100] 3 third substrate [0101] 5 extraction
chamber-forming non-bonded area [0102] 6 extraction chamber [0103]
7, 37, 47, 68a to 68h through-hole [0104] 8 flow channel [0105] 9
mixing chamber-forming non-bonded area [0106] 10 and 20 chip for
analysis of a target substance [0107] 11, 31, 41, 51a to 51h, 61a
to 61h, 62a to 62h, and 71 flow channel-forming non-bonded area
[0108] 12a to 12z, and 12a to 12a shutter-forming non-bonded area
[0109] 13 magnet [0110] 14 adapter [0111] 15 injection tube [0112]
16 magnetic particle [0113] 17a, 17b, 17d shutter-forming void
[0114] 18a to 18z, 18a to 18e pressure supply port [0115] 19 mixing
chamber [0116] 30 washing reagent supply portion [0117] 40 PCR
reaction reagent supply portion [0118] 50 PCR amplification portion
[0119] 60 electrophoresis analysis portion [0120] 65a to 65h, 66a
to 66h, 78 waste tank [0121] 70 washing reagent recovery
portion
* * * * *