WO2008041718A1

WO2008041718A1 - Microchip, and microchip electrophoresis device

Info

Publication number: WO2008041718A1
Application number: PCT/JP2007/069340
Authority: WO
Inventors: Hiroaki Misawa; Sumihare Noji
Original assignee: National University Corporation Hokkaido University; The University Of Tokushima
Priority date: 2006-10-04
Filing date: 2007-10-03
Publication date: 2008-04-10
Also published as: JPWO2008041718A1; JP4862115B2

Abstract

Provided is a microchip electrophoresis device capable of dispensing a specific sample component from a sample and recovering the dispensed specific sample component into a pod without being adsorbed by an electrode. A microchip to be used in the microchip electrophoresis device is constituted to include a separate passage (104), a dispensing passage (106) intersecting with the separate passage, a recovery reservoir (pod) (118) communicating with the dispensing passage, and a recovery electrode reservoir (120) communicating with the recovery reservoir and having a recovery electrode. The microchip electrophoresis device separates the specific sample component contained in the sample, through the separate passage, dispenses the specific sample component through the dispensing passage, and recovers the dispensed specific sample component into the recovery reservoir. After this, the microchip electrophoresis device applies a voltage to the recovery reservoir from the recovery electrode so that the sample component in the recovery reservoir may leave the electrode.

Description

Specification

Microchip and microchip electrophoresis apparatus

Technical field

[0001] The present invention relates to a microchip and a microchip electrophoresis apparatus, and more particularly to a microchip and a microchip electrophoresis apparatus for separating a specific sample component from a sample.

Background art

[0002] Electrophoresis is a separation method that utilizes a phenomenon in which ions are directed to a pole having an opposite charge when a direct current voltage is applied to a solution. Among them, gel electrophoresis separates macromolecules by utilizing the fact that macromolecules such as DNA and proteins are blocked by carrier molecules (gels such as agarose and polyacrylamide) and move with increasing molecular weight. It is a method. Gel electrophoresis has been used to determine the base sequence of genomic DNA, such as in the Human Genome Project, because DNA can be separated by the difference in the number of bases.

[0003] In 2003, it was declared that the decoding of the base sequence of the human genome was completed, and it entered the so-called post-genomic era. At present, it is an important issue to clarify genetic information and disease-related information possessed by a base sequence that is more than just determining the DNA base sequence. In order to accomplish this task, it is necessary to take out (sort) the desired DNA from the enormous amount of nucleic acids (DNA and RNA) present in the cell.

[0004] Conventionally, in order to fractionate desired DNA, the desired DNA separated by gel electrophoresis is cut out together with the surrounding gel with a cutter or a scalpel, and the desired DNA is extracted from the cut out gel. Has been done. This method was cumbersome for researchers who had a lot of work to do. Therefore, there has been a demand for the introduction of a high-speed and automated sorting device that is easy for researchers.

[0005] A microchip electrophoresis apparatus disclosed in Patent Document 1 is disclosed as such a sorting apparatus. The microchip electrophoresis apparatus of Patent Document 1 includes a microchip having a flow path shown in FIG. 1, and controls a voltage applied to each electrode of the microchip to control a desired sample from a sample containing a plurality of DNAs. Sample components (specific DNA) The power S

[0006] The operation of the microchip electrophoresis apparatus disclosed in Patent Document 1 will be described with reference to FIG. First, a sample is placed in the pod 2a, and the sample is migrated in the introduction migration groove 5. When the desired band reaches the intersection between the introduction migration groove 5 and the fractionation migration groove 6, the voltage applied to each electrode is switched, and the desired band is introduced into the fractionation groove 6a (FIG. 1A: Introduction process). Next, the introduced band is migrated in the fractionation groove 6a, and the sample is fractionated (separated) into sample components (FIG. 1B: fractionation treatment). When the desired band reaches the intersection between the fractionation groove 6a and the sorting groove 6b, the voltage applied to each electrode is switched, and the desired band is sorted into the sorting groove 6b. Finally, collect the collected bands in pods 2f to 2m (Figure 1C: Sorting process).

[0007] As described above, the microchip electrophoresis apparatus of Patent Document 1 can sort out desired DNA without a researcher performing complicated operations such as cutting out the gel or extracting from the gel.

Patent Document 1: JP 2002-310992 A

Disclosure of the invention

Problems to be solved by the invention

[0008] However, the microchip electrophoresis apparatus of Patent Document 1 has a problem that the collected sample components are adsorbed to the electrodes in the pod.

That is, in the microchip electrophoresis apparatus of Patent Document 1, since the voltage is applied to the electrode in the pod for a certain period of time after the sample component is collected in the pod, the collected sample component is It is adsorbed by the inner electrode. Thus, if the sample component is adsorbed on the electrode, it becomes difficult to extract the collected sample component from the pod using a micropipette or the like. In addition, if the electrode is adsorbed and contaminated by the sample component, the ability of the electrode to draw the sample component will decrease.

[0010] An object of the present invention is to collect a specific sample component from a sample, adsorb the collected specific sample component on an electrode, and collect the microchip and the microchip electricity that can be recovered in a state. It is to provide a swimming device.

Means for solving the problem

[0011] The microchip of the present invention includes a separation channel, a sorting channel intersecting with the separation channel, and A separation reservoir that communicates with the sorting channel, the separation channel separates a specific component contained in a sample migrated therein, the sorting channel separates the specific component, and The collection reservoir is a microchip that collects the specific component, and includes a collection electrode reservoir having a collection electrode that communicates with the collection reservoir and applies a voltage to the collection reservoir.

[0012] The microchip electrophoresis apparatus of the present invention includes a separation channel having electrodes at both ends, a sorting channel crossing the separation channel, a collection reservoir communicating with the sorting channel and having an electrode, A microchip having a recovery electrode reservoir that communicates with the recovery reservoir and applies a voltage to the recovery reservoir; and the specific component that migrates in the separation channel of the microchip. A separation channel detection unit for optical detection, a collection reservoir detection unit for optical detection of the specific component collected in the collection reservoir of the microchip, the separation channel detection unit and the collection reservoir detection A control unit that determines a voltage to be applied to the electrode of the microchip based on a detection result of the unit, and a voltage that applies a voltage to the electrode of the microchip based on the determination by the control unit. And a application unit.

The invention's effect

[0013] According to the present invention, it is possible to adsorb the collected specific sample component to the electrode and recover it in the recovery reservoir in a state of! / ,! Therefore, according to the present invention, it is possible to quickly collect a specific sample component from a sample and collect the sample component that has been collected in a state where it can be easily taken out with a micropipette or the like.

Brief Description of Drawings

FIG. 1 is a diagram for explaining the operation of a conventional microchip electrophoresis apparatus. (A) is a schematic diagram of a microchip during introduction processing, and (B) is a microchip during fractionation processing. (C) is a schematic diagram of the microchip during the sorting process.

2 is a diagram showing a microchip according to Embodiment 1 of the present invention, (A) is a plan view of the microchip, (B) is a partially enlarged plan view of the microchip, and (C) is a microchip. FIG. 4 is an enlarged sectional view of a part of the chip.

FIG. 3 is a schematic view showing the structure of a microchip electrophoresis apparatus of the present invention. FIG. 4 is an enlarged plan view of an intersection of a separation channel and a sorting channel.

FIG. 5 is a flowchart for explaining the operation of the microchip electrophoresis apparatus according to the first embodiment of the present invention.

FIG. 6 is a schematic diagram for explaining an operation of collecting sample components of the microchip electrophoresis apparatus according to the first embodiment of the present invention.

FIG. 7 is a plan view of a microchip according to a second embodiment of the present invention.

FIG. 8 is a flowchart for explaining the operation of the microchip electrophoresis apparatus according to the second embodiment of the present invention.

FIG. 9 is a schematic diagram for explaining an operation of collecting sample components of the microchip electrophoresis apparatus according to the second embodiment of the present invention.

FIG. 10 is a photograph of the microchip used in the example, (A) is a photograph showing the intersection of the introduction channel and the separation channel, and (B) is a photograph of the intersection of the separation channel and the sorting channel. It is a photograph shown.

FIG. 11 is a photograph showing the intersection of the introduction channel and the separation channel during the introduction process.

[Fig. 12] Fluorescent photographs showing the intersections of the separation flow path and the preparative flow path, (A) to (C) are fluorescent photographs during blocking processing, and (D) to (F) are during preparative processing. Fluorescent photographs (G) to (I) are fluorescent photographs during the recovery process.

FIG. 13 is a fluorescent photograph showing the intersection of a separation channel and a sorting channel when no blocking treatment is performed.

FIG. 14 is a chromatogram of fluorescence intensity near the intersection of the separation channel and the sorting channel.

FIG. 15 is a fluorescent photograph of the area around the nanochannel in the collection reservoir when a voltage is applied to the collection electrode reservoir.

BEST MODE FOR CARRYING OUT THE INVENTION

[0015] 1. Microchip of the present invention

The microchip of the present invention is a microchip having a separation channel provided with electrodes at both ends, a sorting channel intersecting with the separation channel, and a collection reservoir communicating with the sorting channel and having an electrode. And a collection electrode reservoir further comprising a collection electrode in communication with the collection reservoir.

The separation channel is provided with electrodes at both ends, and the introduced sample is electrophoresed. A flow path for separating a specific sample component from a sample. The separation channel is preferably filled with a buffer solution, and more preferably filled with a carrier. The carrier is not particularly limited, and for example, agarose gel or polyacrylamide gel can be used. If the separation channel detector described later detects fluorescence, a fluorescent dye may be added to the carrier or buffer solution! The thickness of the separation channel (width x depth) may be set appropriately according to the type and amount of the sample. For example, the width of the separation channel is preferably in the range of 50 to; lOO ^ m, and more preferably in the range of 70 to 100 m. The length of the separation channel is not particularly limited. For example, the length from the point where the sample is introduced to the intersection with the sorting channel may be about 65 mm.

[0017] The sorting channel is a channel that sorts a specific sample component migrated in the separation channel at the intersection of the separation channel and the sorting channel. The sorting flow path is preferably filled with a buffer solution S, and may further be filled with a carrier. The thickness of the sorting channel is not particularly limited! /, But is preferably narrower than the separation channel from the viewpoint of increasing the resolution of the sorting process described later. For example, the width of the sorting channel is preferably in the range of 25 to 75 111, and more preferably in the range of 25 to 60 111. The length of the sorting channel (the distance from the intersection of the separation channel and the sorting channel to the collection reservoir) is not particularly limited, but may be, for example, about 5 to 10 mm.

[0018] The collection reservoir has a pod filled with a buffer solution inside and an electrode for applying a voltage to the sorting channel, and a specific sample component sorted in the sorting channel is stored in the pod. To collect. The size of the pod is not particularly limited as long as sample components can be collected, but a depth of about 1 mm is preferred, with a diameter of about 500 ^ 111-4 mm being preferred. The material of the electrode is not particularly limited as long as a voltage can be applied to the preparative flow path, but gold or platinum is particularly preferred, which is preferably a metal that is difficult to bend such as gold, platinum, copper, and aluminum!

[0019] The collection electrode reservoir has a pod filled with a buffer solution therein, and a collection electrode for applying a voltage to the collection reservoir. The collection electrode reservoir preferably communicates with the collection reservoir through a nanochannel that allows water molecules to pass through but does not allow sample components to pass through. The material of the nanochannel is not particularly limited as long as it allows water molecules to pass through but does not allow sample components to pass through, but for example, it is porous glass made of sodium silicate. Any porous material can be used. The size of the pod and the material of the collection electrode are not particularly limited, and may be the same as the collection reservoir. The length of the nanochannel and the distance between the collection reservoir and the collection electrode reservoir are not particularly limited as long as the collection electrode reservoir can apply a voltage to the collection reservoir, and depends on the material of the nanochannel. What is necessary is just to adjust suitably. For example, when sodium silicate porous glass is used for the nanochannel, the length of the nanochannel is 5 to 10 mm. The distance between the collection reservoir and the collection electrode reservoir is 0.5 to 1 mm. Any degree is acceptable.

The microchip of the present invention can be used in any microchip electrophoresis apparatus, for example, the following microchip electrophoresis apparatus.

[0021] 2. Microchip electrophoresis apparatus of the present invention

The microchip electrophoresis apparatus of the present invention includes the above-described microchip of the present invention, a separation flow path detection unit, a recovery reservoir detection unit, a control unit, and a voltage application unit.

The separation channel detection unit optically detects a sample component that migrates in the separation channel in the vicinity of the intersection of the separation channel and the sorting channel of the microchip. At this time, the separation channel detection unit is migrated in the separation channel near the intersection of the separation channel and the separation channel, which is composed of only the sample components to be migrated in the separation channel! It is also preferable to detect sample components! The method for optically detecting the sample component by the separation channel detection unit is not particularly limited, but for example, fluorescence emitted from the sample component stained with a fluorescent dye may be detected! /. The detection result is output to the control unit.

[0023] The collection reservoir detection unit optically detects a sample component collected in the collection reservoir in the collection reservoir of the microchip. The method for optically detecting the sample component by the collection reservoir detector is not particularly limited, and may be the same method as the separation channel detector. The detection result is output to the control unit.

[0024] The control unit determines a voltage to be applied to each electrode of the microchip. For example, when a specific sample component is separated from the sample by migrating the sample in the separation channel, the control unit applies a voltage of the opposite polarity to the charge of the sample component at the downstream electrode in the separation channel. Decide to apply. In addition, when the separation channel detection unit detects that the desired sample component has reached the intersection of the separation channel and the sorting channel, or the desired sample component is contained in the sorting channel. When migrating, the control unit determines to apply a voltage having a polarity opposite to the charge of the sample component to the electrode in the collection reservoir. In addition, when the collection reservoir detection unit detects that the collected sample component has been collected in the collection reservoir, the control unit detects the charge of the sample component on the electrode in the collection electrode reservoir (collection electrode). Decide to apply a voltage of opposite polarity. The voltage applied to each electrode by the control unit is not particularly limited as long as sample components can be migrated. For example, a voltage of about 150 V / cm may be applied to the flow path for running the sample components. Whether the sample component detected by the separation channel detection unit is a desired sample component is preferably set in advance by the user! /. For example, when it is known in the preliminary experiment that the order in which each sample component in the sample reaches the intersection of the separation channel and the separation channel, the user will be able to “separate the sample component that reaches the intersection second. ”And set it in the control!

The voltage application unit applies a voltage to each electrode of the microchip based on the determination of the control unit. The voltage application unit may be provided with, for example, a power source and a switching switch circuit group having a relace switch circuit.

[0026] Sample components that can be collected and collected by the microchip and microchip electrophoresis apparatus of the present invention are not particularly limited, and examples thereof include DNA, RNA, protein, and cells.

[0027] Hereinafter, a flow in which the microchip electrophoresis apparatus of the present invention separates a desired sample component from a sample and collects it in a collection reservoir will be described.

First, the voltage application unit applies a voltage to the separation channel of the microchip based on the determination of the control unit, and the sample introduced into the separation channel is separated from the introduction point to the separation channel. Electrophoresis toward the intersection of the and the separation flow path. Thus, the sample is migrated toward the intersection of the separation channel and the sorting channel while being separated into each sample component by the molecular sieve effect (separation process).

[0029] Next, when the separation channel detection unit detects that a desired sample component has arrived at the intersection of the separation channel and the sorting channel, the voltage application unit determines whether the micro-channel is based on the determination of the control unit. A voltage is applied to the tip separation channel. As a result, the desired sample component is taken into the sorting channel (sorting process) and collected in the collecting reservoir (collecting process). Note that the separation channel detector is not desired! / Even if it is detected that the sample component has reached the intersection of the separation channel and the sorting channel, the voltage Since the applying unit does not apply a voltage to the sorting channel based on the determination of the control unit, the separating process is continued without performing the sorting process. Therefore, undesired sample components are further migrated toward the electrode downstream of the separation channel without being taken into the sorting channel.

[0030] Next, when the collection reservoir detection unit detects that a desired sample component is collected in the collection reservoir, the voltage application unit applies a voltage to the collection electrode of the microchip based on the determination of the control unit. Is applied. As a result, the sample component adsorbed on the electrode in the collection reservoir moves away from the electrode in the collection reservoir and further migrates toward the collection electrode reservoir. The sample component is blocked from migration by the nanochannel between the collection reservoir and the collection electrode reservoir, so it eventually remains in the area surrounding the nanochannel in the collection reservoir.

As described above, the microchip electrophoresis apparatus of the present invention separates a desired sample component from a sample, and collects the collected sample component in a collection reservoir in a state where it can be easily taken out with a micropipette or the like. Can do.

[0032] 3. Introduction channel

The microchip of the present invention preferably further includes a sample reservoir having electrodes, and an introduction channel that communicates with the sample reservoir and intersects the separation channel.

[0033] The sample reservoir includes a pod into which a sample is injected, and an electrode for applying a voltage to the sorting channel. The size of the pod is not particularly limited as long as a desired amount of the sample component can be injected, but a depth of about 1 mm is preferable, and a diameter of about 500 111 to 4111 111 is preferable. The material of the electrode is not particularly limited as long as a voltage can be applied to the introduction flow path, and may be the same as the material of the electrode of the recovery reservoir.

[0034] The introduction channel is provided with electrodes (one of which is an electrode in the sample reservoir) at both ends, and the sample injected into the sample reservoir is transferred from the intersection of the introduction channel and the separation channel to the separation channel. This is a flow channel to be introduced. The introduction channel is preferably filled with a buffer solution inside, and may further be filled with a carrier. The thickness of the introduction channel (width X depth) is not particularly limited, but may be, for example, about 90 m width x 25 Hm depth. The length of the introduction channel is not particularly limited. For example, the length from the communication point with the sample reservoir to the intersection with the separation channel may be about 5 mm. [0035] The sample reservoir and the introduction channel are used for introducing the sample into the separation channel (introduction process) performed before the separation process. Hereinafter, the flow of the introduction process will be described.

[0036] First, a sample containing a desired sample component is injected into a sample reservoir. Next, the voltage application unit applies a voltage to the introduction flow path to cause the sample to migrate from the sample reservoir toward the intersection of the introduction flow path and the separation flow path. After the sample reaches the intersection of the introduction channel and the separation channel, the voltage application unit introduces a part of the sample into the separation channel by applying a voltage to the separation channel for a short time.

[0037] By using the sample reservoir and the introduction channel, the sample can be introduced into the separation channel while preventing diffusion and leakage of the sample. Therefore, since a clear sample plug can be formed in the separation channel, it is possible to increase the resolution of subsequent separation processing and sorting processing.

[0038] 4. Block electrode

The microchip of the present invention preferably further has a block electrode.

[0039] The block electrode is an electrode that is provided at a position communicating with the sorting channel and applies a voltage to the sorting channel. The material of the block electrode is not particularly limited as long as a voltage can be applied to the introduction flow path, and may be the same as the material of the electrode of the recovery reservoir.

[0040] The block electrode is used during the separation process. When the separation channel detection unit detects that a sample component other than a specific sample component has reached the intersection of the separation channel and the sorting channel, the control unit has the same polarity as the charge of the sample component on the block electrode Decide to apply a voltage of. The voltage application unit applies a voltage to the block electrode based on the determination of the control unit (blocking process).

[0041] By using the block electrode, it is possible to prevent a sample component other than the specific sample component from entering the sorting flow path. Therefore, the sorting channel can be prevented from being stained by sample components other than a specific sample component.

[0042] Further, the microchip of the present invention preferably has two or more block electrodes. At this time, it is more preferable that the block electrodes are disposed on both lateral sides of the separation channel. When the sorting channel is on both sides of the separation channel (for example, when the separation channel and the sorting channel cross in a cross), one block electrode has a voltage across the sorting channel on one side. And apply the other By applying a voltage to the sorting channel on the other side, the lock electrode can prevent the sorting channels on both sides from being contaminated by sample components other than the specific sample component.

[0043] When the microchip of the present invention has a block electrode, the thickness of the sorting channel is preferably thinner than the separation channel from the viewpoint of increasing the resolution of the blocking process. For example, the width of the sorting channel is preferably in the range of 25 to 75 111, more preferably in the range of 25-60 ^ 111 (7) | g.

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

[0045] (Embodiment 1)

FIG. 2A is a diagram showing a configuration of a microchip used in the microchip electrophoresis apparatus according to Embodiment 1 of the present invention.

In FIG. 2A, the microchip 100 according to the present embodiment includes an introduction channel 102, a separation channel 104, a sorting channel 106, two block channels 108a and 108b, a sample reservoir 110, and a sample discharge. It has a reservoir 112, a buffer reservoir 114, a buffer discharge reservoir 116, a collection risano 118, a collection electrode reservoir 120, and two block electrode reservoirs 122a and 122b. For example, the size of the microchip 100 should be about 100 mm (horizontal direction in the figure) X 30 mm (vertical direction in the figure)! /.

[0047] The introduction channel 102 intersects with the separation channel 104, and is a channel provided with a sample reservoir 110 and a sample discharge reservoir 112 at both ends thereof. For example, the introduction flow path 102 may be a flow path having a width of 90 Hm, a depth of 25 μm, and a length of 10 mm, and intersecting the separation flow path 104 at the midpoint.

[0048] The separation channel 104 intersects with the introduction channel 102 and the sorting channel 106 so as to communicate with each other, and the buffer reservoir 114 and the buffer discharge reservoir 116 are provided at both ends thereof. For example, the separation channel 104 has a width of 90 μm, a depth of 25 μm, and a length of 85 mm. The separation channel 104 intersects the introduction channel 102 at a point 5 mm from the buffer reservoir 114 and It is sufficient to cross the sorting channel 106 at a point 15 mm from the discharge reservoir 116.

[0049] The sorting channel 106 intersects with the separation channel 104 so as to communicate with the separation channel 104, and is provided with a collection reservoir 118 at one end. In addition, the sorting channel 106 includes block channels 108a and 108b. Also communicate. For example, a portion of the sorting channel 106 that is perpendicular to the separation channel 104 (see FIG. 2A) is a channel having a width of 55 111, a depth of 25 111, and a length of 10 mm. It is sufficient to cross the road 104. In addition, a portion of the sorting channel 106 that is parallel to the separation channel 104 (see FIG. 2A) may be, for example, a channel having a width of 90 Hm, a depth of 25 μm, and a length of 5 mm. ! /

[0050] The block channels 108a and 108b are channels that communicate with the sorting channel 106 at one end, and are channels in which the block electrode reservoirs 122a and 122b are provided at the other end. For example, the block channels 108a and 108b may be channels having a width of 90 mm 111, a depth of 25 mm, and a length of 20 mm.

[0051] Each of the Sampno Relizano 110, the sample discharge reservoir 112, the buffer reservoir 114, the buffer discharge reservoir 116, the collection reservoir 118, the collection electrode reservoir 120, and the block electrode reservoirs 122a and 122b has a buffer solution ( It has a pod for storing a buffer) and a sample (sample), and an electrode (not shown! /) For applying a voltage to each channel. Here, the electrode of the collection electrode reservoir 120 is referred to as a “collection electrode”. Similarly, the electrodes respectively included in the block electrode reservoirs 122a and 122b are referred to as “block electrodes”. In the present specification, applying a voltage to the electrode in the reservoir A may be referred to as “applying a voltage to the reservoir A”. For example, the size of the pods of the reservoirs 110 to 122 may be about 3 to 4 mm in diameter and about 1 mm in depth.

FIG. 2B is a partially enlarged plan view of the microchip 100 showing the collection reservoir 118 and the collection electrode reservoir 120. FIG. 2C is a partially enlarged sectional view of the microchip 100 showing the collection reservoir 118 and the collection electrode reservoir 120. For convenience of explanation, electrodes are not shown.

[0053] In FIG. 2B and FIG. 2C, the collection electrode reservoir 120 is formed adjacent to the collection reservoir 118, and passes through the nanochannel 124 through which water molecules pass but sample components do not pass. Communicate with 118.

FIG. 3 is a diagram showing a configuration of the microchip electrophoresis apparatus according to the first embodiment of the present invention.

In FIG. 3, the microchip electrophoresis apparatus 200 according to the present embodiment includes the microchip 100, the separation channel detection unit 210, the recovery reservoir detection unit 220, and the control unit shown in FIG. 230 and a voltage application unit 240.

The separation channel detection unit 210 detects the fluorescence emitted from the sample component in the vicinity of the intersection between the separation channel 104 and the sorting channel 106 of the microchip 100. FIG. 4 is a schematic diagram showing the vicinity of the intersection between the separation channel 104 and the sorting channel 106. In FIG. 4, the left side is the buffer reservoir 114 side, the right side is the buffer discharge reservoir 116 side, the upper side is the block electrode reservoir 122a and recovery reservoir 118 side, and the lower side is the block electrode reservoir 122b side. For example, the separation channel detection unit 210 detects fluorescence at three points a to _C in FIG. 4 to separate the sample component (point a) that has migrated in the separation channel 104 into the separation channel. It is possible to distinguish and detect the sample component (point b) that has not been sampled and the sample component (point c) separated in the sorting channel 106. The detection result is output to the control unit 230.

For example, as shown in FIG. 3, the separation channel detection unit 210 includes a mercury lamp 211 as an excitation light source, an excitation filter 212 that separates excitation light, and guides the excitation light in the direction of the sample component. What is necessary is just to comprise the dichroic mirror 213 which guides the fluorescence emitted from the components in the direction of the CCD camera 215, the absorption filter 214 for separating the fluorescence, and the CCD camera 215 for detecting the fluorescence. The method for emitting fluorescence from the sample component may be appropriately selected according to the sample component. For example, if the sample component is DNA, the sample component may be stained with a DNA-binding fluorescent dye such as ethidium bromide or cyber green.

The collection reservoir detection unit 220 detects fluorescence emitted from the sample components in the collection reservoir 118 of the microchip 100. The collection reservoir detection unit 220 may have the same configuration as that of the separation flow path detection unit 210, for example. The detection result is output to the control unit 230.

The control unit 230 determines a voltage to be applied to the electrodes in each reservoir of the microchip 100. At this time, the control unit 230 determines the position of each sample component in the vicinity of the intersection of the separation channel 104 and the separation channel 106 from the detection result of the separation channel detection unit 210, and whether the sample component should be separated. The voltage to be applied to each electrode of the microchip 100 is determined depending on whether or not. Further, the control unit 230 determines whether or not the sample component collected from the detection result of the collection reservoir detection unit 220 is collected in the collection reservoir 118, and whether or not the sample component is collected in the collection reservoir 118. The voltage applied to each electrode of the microchip 100 is determined accordingly. For example, if the sample component is DNA (negatively charged), the flow path for running the sample component Apply a positive voltage to the downstream electrode and ground the upstream electrode! /.

The voltage application unit 240 applies a voltage to the electrodes in each reservoir of the microchip 100 based on the determination by the control unit 230. For example, as shown in FIG. 3, the voltage application unit 240 may include power supplies 242a to 242c and a switching switch circuit group 244 having a relaced circuit.

Next, using the flowchart shown in FIG. 5 and the schematic diagram of the microchip shown in FIG. 6, the microchip electrophoresis apparatus according to the present embodiment separates and collects a desired sample component. The flow to do will be described.

[0062] First, in step S1000, a sample containing a desired sample component is injected into the sample reservoir 110.

Next, in step S 1100, the sample is migrated from the sample reservoir 110 toward the sample discharge reservoir 112 by applying a voltage to the introduction channel 102, and the sample is introduced into the introduction channel 102 and the separation channel 104. After reaching the crossing point, a part of the sample is introduced into the separation channel 104 by applying a voltage to the separation channel 104 for a short time (introduction process). For example, when the sample component is DNA, in order to migrate the sample from the sample reservoir 110 to the sample discharge reservoir 112, the sample reservoir 110 is grounded and a positive voltage is applied to the sample discharge reservoir 112. At this time, the buffer reservoir 114 and the buffer discharge reservoir 116 are also preferably grounded so that contaminants do not enter the separation channel 104. In order to introduce a part of the sample into the separation channel 104 after the sample reaches the intersection of the introduction channel 102 and the separation channel 104, for example, the sample reservoir 110 is grounded and the sample discharge reservoir 112 is grounded. And apply a positive voltage to the buffer drain reservoir 116 for 4 seconds.

Next, in step S 1200, the sample is migrated from the intersection of the introduction flow path 102 and the separation flow path 104 toward the buffer discharge reservoir 116 by applying a voltage to the separation flow path 104. At this time, the sample is separated into each sample component by the molecular sieving effect (separation process). For example, if the sample component is DNA, as shown in Fig. 6A, ground the buffer reservoir 114 (indicated by "-" in the figure) and apply a positive voltage to the notch drain reservoir 116! / (Indicated by “+” in the figure). [0065] Next, in step S1300, when the separation channel detection unit 210 detects that the separated sample component has reached the intersection of the separation channel 104 and the sorting channel 106 (see FIG. 6A). The controller 230 determines whether or not this sample component is a desired sample component. If this sample component is not the desired sample component (S1300: NO), go to step S1400. On the other hand, if this sample component is the desired sample component (S 1300: YES), the process proceeds to step S 1500.

In step S 1400, an undesired sample component is swung toward the buffer discharge reservoir 116. At this time, voltage is applied to the block electrode reservoirs 122a and 122b (block electrodes) to prevent the sample component from entering the sorting channel 106 (blocking process). For example, when the sample component is DNA, if the separation channel detection unit 210 detects that the fluorescence intensity at the point b in FIG. 4 has risen above a predetermined threshold, the block electrode reservoirs 122a and 122b can be grounded. Good (see Figure 6B). When this sample component completely passes through the intersection of the separation channel 104 and the separation channel 106, the block electrode reservoirs 122a and 122b are returned to the open state (not shown in the figure), and the process returns to step S1200 (FIG. 6C). reference).

[0067] In step S1500, a desired sample component is taken into the sorting channel 106 by applying a voltage to the sorting channel 106 (sorting process). For example, when the sample component is DNA, the buffer reservoir 114 is grounded, the buffer discharge reservoir 116 is opened, and a positive voltage is applied to the collection reservoir 118 as shown in FIG. 6D.

Next, in step S 1600, the collected sample component is collected in the collection reservoir 118 (collection process). For example, if the sample component is DNA, the buffer reservoir 114 is opened, the block electrode reservoirs 122a and 122b are grounded, and a positive voltage is applied to the collection reservoir 118 (see FIG. 6E), so that the collected sample components Can be recovered in the recovery reservoir 118 (see Figure 6F).

[0069] Next, in step S1700, when the collection reservoir detection unit 220 detects that the sample components collected in the collection reservoir 118 have been collected, a voltage is applied to the collection electrode reservoir 120. Then, the sample component is separated from the electrode in the collection reservoir 118, and this flow is finished. For example, if the collected sample component is DNA, as shown in FIG. 6G, the block electrode reservoirs 122a and 122b are grounded, the collection reservoir 118 is opened, and a positive voltage is applied to the collection electrode reservoir 120. do it. By doing this, The sample component adsorbed on the electrode in the collection reservoir 118 moves away from the electrode and further migrates toward the collection electrode reservoir 120. Since sample components are prevented from migrating by the nanochannel 124, the sample component will eventually end up in the region around the nanochannel 124 in the collection reservoir 118.

As described above, according to the present embodiment, the collected sample component is prevented from adsorbing to the electrode in the collection reservoir, and the collected sample component is collected in the region around the nanochannel in the collection reservoir. Therefore, the sample components in the collection reservoir can be easily removed using a micropipette.

[0071] Further, according to the present embodiment, since the block electrode applies a voltage to the sorting flow path, it is possible to prevent a desired layer and band from entering the sorting flow path. The ability to sort and collect components without contamination is possible.

[0072] Further, according to the present embodiment, the voltage to be applied to each electrode of the microchip is determined by the control unit based on the detection results of the separation channel detection unit and the recovery reservoir detection unit. Sample components can be accurately sorted by automatic control without any work.

[0073] (Embodiment 2)

In the first embodiment, an example in which there is one recovery reservoir is shown. Embodiment 2 shows an example in which there are a plurality of recovery reservoirs. The same components as those of the microchip and the microchip electrophoresis apparatus according to the first embodiment are denoted by the same reference numerals, and description of overlapping portions is omitted.

FIG. 7 is a diagram showing a configuration of a microchip according to Embodiment 2 of the present invention.

In FIG. 7, the microchip 300 of the present embodiment includes an introduction channel 102, a separation channel 104, a sorting channel 302, two block channels 108a and 108b, a sample reservoir 110, and a sample. It has a discharge reservoir 112, a buffer reservoir 114, a buffer discharge reservoir 116, a plurality of recovery reservoirs 304a to 304d, a plurality of recovery electrode reservoirs 306a to 306d, and two block electrode reservoirs 122a and 122b. The components other than the sorting channel 302 are the same as those in the first embodiment.

The sorting channel 302 intersects with the separation channel 104 and communicates with the separation channel 104. Is a flow path that branches into a plurality of sides on both sides. Recovered Lisano 304a to 304d forces S are provided at the branch ends of the sorting flow path 302, respectively. For convenience of explanation, the sorting flow paths 302 communicating with the collection reservoirs 304a to 304d may be referred to as branch flow paths 302a to 302d, respectively. Further, the sorting channel 302 communicates with the block channels 108a and 108b. For example, a portion of the sorting channel 302 that is perpendicular to the separation channel 104 (see FIG. 7) is a channel having a width of 55 m, a depth of 25 ^ m, and a length of 10 mm, and an intermediate point (1 / 2) crosses the separation channel 104 and divides at both ends, 1/4 and 3/4. The branch channels 302a and 302d are, for example, 90 μm wide, 25 μm deep, and 5 mm long, and the branch channels 302b and 302c are 90 mm wide. The channel should be 111, 25 mm deep and 3 mm long.

[0077] A microchip electrophoresis apparatus 200 according to Embodiment 2 includes a microchip 300, a separation channel detection unit 210, a plurality of collection reservoir detection units 220a to 220d, a control unit 230, and a voltage application shown in FIG. Part 240 is provided. Components other than the microchip 300 are the same as those in the first embodiment.

[0078] Next, using the flowchart shown in FIG. 8 and the schematic diagram of the microchip shown in FIG. 9, the flow in which the microchip electrophoresis apparatus according to the present embodiment sorts a desired sample component will be described. To do. Steps S1000 to S1200 (separation process: see FIG. 9A) and step S1400 (blocking process: see FIGS. 9B and 9C) are the same as the steps in the flowchart of FIG.

[0079] In step S2000, when the separation channel detector 210 optically detects that the separated sample component has reached the intersection of the separation channel 104 and the sorting channel 302 (see FIG. 9A). The control unit 230 determines whether this sample component is a desired sample component. In Embodiment 2, the separation channel detection unit 210 detects fluorescence at four points a to d near the intersection of the separation channel and the sorting channel shown in FIG. To do. If this sample component is not the desired sample component! / (S2000: NO), go to step S1400. On the other hand, if this sample component is the desired sample component (S2000: YES), the process proceeds to step S2100.

In step S 2100, a desired sample component is taken into the sorting channel 302 by applying a voltage to the sorting channel 106 (sorting process). For example, transfer the desired DNA to the collection reservoir 304a. To achieve this, as shown in FIG. 9D, the buffer discharge reservoir 116 is opened and a positive voltage is applied to the collection reservoir 304a.

[0081] Next, in step S2200, the collected sample components are collected in one of the collection reservoirs 304a to 304d (collection process). At this time, the plurality of recovery reservoirs are used from reservoirs far from the separation channel 104. That is, in the case of the microchip shown in FIG. 7, the sample components are collected in the order of the collection risano 304a, the collection reservoir 304d, the collection reservoir 304b, and the collection reservoir 304c. For example, to collect desired DNA in the collection reservoir 304a, open the buffer reservoir 114, ground the block electrode reservoirs 122a and 122b, and apply a positive voltage to the collection reservoir 304a (see FIG. 9E). The collected sample components can be collected in the collection reservoir 304a (see FIG. 9G). Further, when the collected sample component reaches the branch flow path 302a, the branch flow path 302b is cleaned by applying a voltage to the branch flow path 302b located inside the branch flow path 302a (washing). processing). For example, when the sample component is DNA, the recovery reservoir 304b may be grounded as shown in FIG. 9F in order to wash the branch channel 302b. Thus, the sample component that has entered the branch channel 302b during the recovery process can be discharged from the branch channel 302b (see FIGS. 9E and 9F).

Next, in step S2300, when the collection reservoir detector 220a detects that the sample components collected in the collection reservoir 304a are collected, a voltage is applied to the collection electrode reservoir 306a for a predetermined time. As a result, the sample component is separated from the electrode in the collection reservoir 304a. For example, if the collected sample component is DNA, as shown in Fig. 9H, ground the block electrode reservoirs 122a and 122b, open the collection reservoir 304a, and apply a positive voltage to the collection electrode reservoir 306a. do it. By doing so, the sample component adsorbed on the electrode in the collection reservoir 304a is separated from the electrode and further migrates in the direction of the collection electrode reservoir 306a. Since the sample component is prevented from migrating by the nanochannel, it finally remains in the region around the nanochannel in the collection reservoir 304a.

Next, in step S2400, control unit 230 determines whether or not to end this flow.

For example, when all collection reservoirs 304a-304d have collected sample components, all desired sample components have been collected in collection reservoirs 304a-304d, or all When the material component passes through the intersection of the separation channel 104 and the sorting channel 302, it is determined that this flow is finished. If it is determined not to end this flow (S2400: NO), the setting of each electrode is returned to the state of step S1200, and the process returns to step S1200. On the other hand, if it is determined that this flow is to be terminated (S2400: YES), this flow is terminated.

Thus, according to the present embodiment, in addition to the effects of the first embodiment, a plurality of sample components derived from one sample can be collected in different collection reservoirs in one microchip. Touch with force S.

[0085] Further, according to the present embodiment, it is possible to prevent the unused branch flow path from being contaminated by performing the cleaning process, so that the desired sample component can be separated and collected without being contaminated. Is possible.

[0086] Further, according to the present embodiment, the electrodes are arranged symmetrically with respect to the separation channel! /, So that the structure of the electric circuit and the switching operation of voltage application are prevented from being complicated. Doing with the power S

Note that the configuration of the microchip in each embodiment of the present invention is not limited to that described in each of the above embodiments, and the shape and number of each flow path, the number of reservoirs, and the like are changed. Also good.

Example

[0088] The present invention will be further described below with reference to examples. It should be noted that the scope of the present invention is not construed as being limited by this example.

[0089] In this example, an example in which 20 bp DNA (specific sample component) was fractionated from an lObp DNA ladder (sample) using the microchip electrophoresis apparatus including the microchip of Embodiment 1 is shown.

[0090] The microchip used in this example was made of glass having the flow path shown in FIG. 2A. The introduction channel and the separation channel were 90 m wide x 20 m deep. On the other hand, the sorting channel is 50 m wide and 20 m deep to improve sorting resolution. Fig. 10A shows a photomicrograph of the intersection of the introduction channel and the separation channel, and Fig. 10B shows a photomicrograph of the intersection of the separation channel and the sorting channel. In FIG. 10A, “1” indicates the buffer reservoir side, “2” indicates the sample reservoir side, and “4” indicates the sample discharge reservoir side. Figure 10B “3” indicates the buffer discharge reservoir side, “0” indicates the side without the recovery reservoir, and “8” indicates the side with the recovery reservoir. The introduction channel was 10 mm long and crossed the separation channel at the midpoint. The separation channel was 85 mm long, intersecting the inlet channel at a point 5 mm from the buffer reservoir, and intersecting the preparatory channel at a point 15 mm from the buffer discharge reservoir. The part of the sorting channel that is perpendicular to the separation channel is 10 mm long and intersects the separation channel at the midpoint. The part of the sorting channel parallel to the separation channel was 10 mm long. The block channel was 20 mm long.

[0091] The microchip used in this example was manufactured by bonding a glass plate with engraved channel grooves and a glass plate with holes for reservoirs (pods), using sodium silicate as an adhesive layer. did. Specifically, first, using a general photolithography technique and a wet etching technique, a glass plate with engraved channel grooves and a glass plate with holes for reservoirs (pods) were produced. Next, a thin film of sodium silicate was formed on one glass plate by spin-coating a 0.1M sodium silicate aqueous solution. The other glass plate was placed on the sodium silicate thin film and heated at 200 ° C. By doing so, the sodium silicate became a porous glass, and the two glass plates were bonded to each other. The formed sodium silicate porous glass also functions as a nanochannel between the collection reservoir and the collection electrode reservoir, as shown in FIG. 2C. At this time, the length of the nanochannel (“a” in FIG. 2C) was 10 m.

[0092] A polydimethylacrylamide gel containing a DNA-binding fluorescent dye cyber green and a buffer solution were previously introduced into the flow path.

[0093] The separation channel detection unit and the recovery reservoir detection unit are an inverted system microscope (1X70: equipped with a mercury lamp, an excitation filter (460 to 490 nm), a dichroic mirror (505 nm), and an absorption filter (510 to 550 nm). Olympus) and CCD camera (ORCA-EG: Hamamatsu Photonics). The separation channel detector was set to monitor the change in fluorescence intensity at each point A to C shown in FIG. 10B.

[0094] The voltage switching by the control unit and the voltage application unit was set as follows. The separation process was set so that the buffer reservoir was grounded, a voltage of +1300 V was applied to the buffer discharge reservoir, and the other reservoirs were opened (see Figure 6A). Blocking process The two block electrode reservoirs were set to ground when the fluorescence intensity at point B rose above a certain threshold (see Fig. 6B). In addition, when the fluorescence intensity at point B fell below a predetermined threshold, the two block electrode reservoirs were set back to open (see FIG. 6C). The sorting process was set to open the buffer discharge reservoir and apply a voltage of +1300 V to the collection reservoir when the fluorescence intensity at point A rose above a predetermined threshold (see Figure 6D). The collection process was set to open the buffer reservoir and ground the two block electrode reservoirs when the fluorescence intensity at point C fell below a predetermined threshold (see Figure 9E and Figure 9F). When the fluorescence intensity in the collection reservoir rose above a predetermined threshold, the collection reservoir was opened and a voltage of +1300 V was applied to the collection electrode reservoir (see FIG. 9G).

[0095] (Introduction process)

After the sample was injected into the sample reservoir, an introduction process was performed. First, the sample reservoir, the buffer reservoir, and the buffer discharge reservoir were grounded, and a voltage of +300 V was applied to the sample discharge reservoir. As a result, the sample started to migrate from the sample reservoir toward the sample discharge reservoir. Next, when the sample reaches the intersection of the introduction channel and the separation channel, the sample reservoir is grounded, + 300V voltage is applied to the sample discharge reservoir, and + 1300V voltage is applied simultaneously to the sample discharge reservoir for 4 seconds. And a part of the sample was introduced into the separation channel.

FIG. 11 is a fluorescent photograph near the intersection when the sample reaches the intersection between the introduction channel and the separation channel. In FIG. 11, “1” indicates the buffer reservoir side, “2” indicates the sample reservoir side, “3” indicates the buffer discharge reservoir side, and “4” indicates the sample discharge reservoir side. Fluorescently stained sample (lObpDNA ladder)

[0097] (Separation process)

After introducing the sample into the separation channel, electrophoresis by automatic control was started. First, the buffer reservoir was grounded by the control unit and the voltage application unit, and a voltage of +1300 V was applied to the buffer discharge reservoir (see FIG. 6A). Thus, the introduced sample started to migrate toward the buffer discharge reservoir. Sample in a polydimethylacrylamide gel Was separated into multiple bands (10 bp DNA, 20 bp DNA,...).

[0098] (Blocking process)

The multiple bands continued to migrate toward the buffer discharge reservoir, and the force with a small number of bases reached the intersection of the separation channel and the sorting channel in order. Since the first reached band (10 bp DNA) was not the desired DNA (20 bp DNA), blocking treatment was performed. That is, when the fluorescence intensity at point B rises above a predetermined threshold, the two block electrode reservoirs were grounded. As a result, this band (10 bp DNA) migrated toward the buffer discharge reservoir without entering the sorting channel (see FIG. 6B). When the fluorescence intensity at point B fell below a predetermined threshold, the two block electrode reservoirs were returned to the open state and electrophoresis continued (see FIG. 6C).

FIG. 12A to FIG. 12C are fluorescent photographs in the vicinity of the intersection of the separation channel and the sorting channel during the blocking process. Fig. 12A is a photo when an undesired band reaches the intersection, Fig. 12B is a photo when it is detected that the fluorescence intensity at point B has risen above a predetermined threshold, and Fig. 12C is a photo when blocking processing is performed. It is. In each photograph, the vertical channel is the separation channel, the horizontal channel is the sorting channel, the upper is the buffer reservoir side, and the lower is the notch discharge reservoir side. From these photographs, it can be seen that by performing the blocking treatment, an undesired band (10 bp DNA) does not enter the sorting channel.

[0100] FIG. 13 is a fluorescent photograph in the vicinity of the intersection of the separation channel and the sorting channel when a blocking process is applied as a comparative example. The direction of each channel is the same as the photograph in FIG. From this photograph, it can be seen that undesired bands (lOObp DNA and lObp DNA) enter the preparative channel without blocking treatment.

[0101] (Preparation processing)

Since the next band reached (20 bp of DNA) was the desired DNA, sorting and collection were performed. That is, when the fluorescence intensity at point A rose above a predetermined threshold, the buffer discharge reservoir was opened and a voltage of +1300 V was applied to the collection reservoir (see FIG. 6D). As a result, the desired band (20 bp DNA) was separated into the sorting channel. [0102] FIGS. 12D to 12F are fluorescence photographs near the intersection of the separation channel and the sorting channel during the sorting process. Fig. 12D is a photograph when it is detected that the fluorescence intensity at point A has risen above a predetermined threshold, Fig. 12E is a photograph when sorting processing is started, and Fig. 12F is a photograph when sorting processing is finished. . The direction of each channel is the same as the photograph in FIG. 12A. From these photographs, it can be seen that the desired band (20 bp DNA) was fractionated by the fractionation process.

[0103] (Recovery processing)

After the sorting process, a collecting process was performed in order to collect the desired DNA collected in the collecting reservoir. That is, when the fluorescence intensity at point C fell below a predetermined threshold, the buffer reservoir was opened and the two block electrode reservoirs were grounded. As a result, the desired band (20 bp DNA) migrated toward the collection reservoir and was finally collected in the collection reservoir (see FIGS. 6E and 6F).

FIGS. 12G to 121 are fluorescent photographs near the intersection of the separation channel and the sorting channel during the recovery process. 12G is a photograph when the collection process is started, FIG. 12H is a photograph during the collection process, and FIG. 121 is a photograph when the collection process is completed. The direction of each flow path is the same as the photograph in FIG. 12A. From these photographs, it can be seen that the desired band was collected as intended by the collection process.

FIG. 14 is a chromatogram of fluorescence intensity at each point of A to C. The horizontal axis represents time (seconds), and the vertical axis represents fluorescence intensity (arb. Units). The upper row shows the fluorescence intensity at point A, the middle row at point C, and the lower row at point B. In addition, the arrow in the chromatogram (upper part) at point A indicates the start of the desired band collection process, and the arrow in the chromatogram (middle stage) at point C indicates the start of the recovery process for the desired band. Fig. 14 shows that an undesired band (first band) is detected at point A (upper) and then detected at point B (lower), so this band is not sorted. . On the other hand, since the desired band (second band) is detected at point A (upper) and then detected at point C (middle), only this band is separated! I'm worried.

[0106] (Voltage applied to collection electrode reservoir)

After the recovery process, a process was performed to separate the recovered sample components from the electrode in the recovery reservoir. That is, the fluorescence intensity in the collection reservoir is above a predetermined threshold. When raised, the potential of the collection reservoir was opened and a voltage of +1300 V was applied to the collection electrode reservoir. As a result, the sample component (20 bp DNA) in the collection reservoir gathered in the area around the nanochannel in the collection reservoir. (See Figure 6G).

FIG. 15 is a fluorescent photograph of the area around the nanochannel in the collection reservoir when a voltage is applied to the collection electrode reservoir. In FIG. 15, the upper side of the upper dotted line shows the collection reservoir 118, and the lower side of the lower dotted line shows the collection electrode reservoir 120, and what appears white is the sample 400 (20 bp DNA ladder) fluorescently stained with Cyber Green. It is. Thus, it can be seen that when a voltage is applied to the collection electrode reservoir, the sample component (20 bp DNA) in the collection reservoir separates from the electrode in the collection reservoir and collects in the region around the nanochannel.

[0108] The present application (or claiming the priority based on the Japanese Patent Application No. 2006-273377ί, which was filed on October 4, 2006. All the contents described in the specification and drawings of this application are incorporated herein by reference. .

Industrial applicability

[0109] The microchip and the microchip electrophoresis apparatus according to the present invention can separate sample components such as DNA without requiring a complicated work by a researcher, and thus biological materials related to any bioindustry. It is useful for analysis.

Claims

The scope of the claims

[1] having a separation channel, a sorting channel intersecting with the separation channel, and a collection reservoir communicating with the sorting channel,

The separation channel separates a specific component contained in a sample migrated therein, the sorting channel separates the specific component, and the recovery reservoir collects the specific component. A chip,

A microchip having a collection electrode reservoir that communicates with the collection reservoir and has a collection electrode that applies a voltage to the collection reservoir.

[2] The collection electrode reservoir communicates with the collection reservoir via a porous substance.

The microchip according to claim 1.

3. The microchip according to claim 2, wherein the porous material is sodium silicate.

4. The microchip according to claim 1, wherein the sorting channel is narrower than the separation channel.

[5] a sample reservoir;

The microchip according to claim 1, further comprising: an introduction channel that communicates with the sample reservoir and intersects the separation channel.

6. The microchip according to claim 1, further comprising a block electrode that applies a voltage to the sorting channel and suppresses components other than the specific component from entering the sorting channel.

7. The microchip according to claim 6, wherein the number of block electrodes is two or more.

8. The microchip according to claim 7, wherein the block electrodes are arranged on both sides of the separation channel.

[9] A separation channel having electrodes at both ends, a sorting channel crossing the separation channel, communicating with the sorting channel, a collection reservoir having electrodes, and communicating with the collection reservoir, the collection A microphone mouth tip having a collecting electrode reservoir having a collecting electrode for applying a voltage to the reservoir;

A separation channel detection unit for optically detecting a specific component migrated in the separation channel of the microchip;

A collection reservoir detector for optically detecting the specific component collected in the collection reservoir of the microchip; A control unit that determines a voltage to be applied to the electrode of the microchip based on detection results of the separation channel detection unit and the recovery reservoir detection unit;

A voltage applying unit that applies a voltage to the electrodes of the microchip based on the determination of the control unit;

A microchip electrophoresis apparatus having

[10] A separation channel having electrodes at both ends, a sorting channel crossing the separation channel, communicating with the sorting channel, a collection reservoir having electrodes, communicating with the collection reservoir, and the collection reservoir A collecting electrode reservoir having a collecting electrode for applying a voltage to the bar, and a block electrode for applying a voltage to the sorting flow path to prevent components other than the specific component from entering the sorting flow path. A microchip having

A separation channel detector for optically detecting the specific component migrated in the separation channel of the microchip;

A collection reservoir detector for optically detecting the specific component collected in the collection reservoir of the microchip;

A control unit that determines a voltage to be applied to the electrode of the microchip based on detection results of the separation channel detection unit and the recovery reservoir detection unit;

A microchip electrophoresis apparatus having

[11] having a separation channel, a sorting channel intersecting with the separation channel, and a collection reservoir communicating with the sorting channel,

A microchip having a block electrode that applies a voltage to the sorting channel and suppresses components other than the specific component from entering the sorting channel.

12. The microchip according to claim 11, wherein the sorting channel is narrower than the separation channel.

[13] A separation channel having electrodes at both ends, a fractionation channel intersecting with the separation channel, and the fractionation flow A microchip having a collection reservoir that has an electrode in communication with the channel, and a block electrode that prevents a component other than a specific component from entering the sorting channel by applying a voltage to the sorting channel;

A microchip electrophoresis apparatus having