JP4862115B2 - Microchip and microchip electrophoresis apparatus - Google Patents

Description

  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.

  Electrophoresis is a separation method that utilizes the phenomenon that ions migrate toward poles having opposite charges when a direct current voltage is applied to a solution. Among them, gel electrophoresis is a method for separating macromolecules by utilizing the fact that macromolecules such as DNA and proteins are obstructed by carrier molecules (gels such as agarose and polyacrylamide) and that the larger the molecular weight, the more difficult it is to move. It is. Gel electrophoresis has been used to determine the base sequence of genomic DNA in the Human Genome Project and the like because DNA can be separated by the difference in the number of bases.

  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 important not only to determine the base sequence of DNA, but also to clarify the genetic information possessed by the base sequence and information related to diseases. 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.

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

  As such a sorting apparatus, a microchip electrophoresis apparatus disclosed in Patent Document 1 is disclosed. 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 one from a sample containing a plurality of DNAs. Sample components (specific DNA) can be separated.

  The operation of the microchip electrophoresis apparatus disclosed in Patent Document 1 will be described with reference to FIG. First, a sample is put in the pod 2 a and the sample is migrated in the introduction migration groove 5. When the desired band reaches the intersection of the introduction electrophoresis groove 5 and the fractionation electrophoresis groove 6, the voltage applied to each electrode is switched to introduce the desired band 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 process). When the desired band reaches the intersection of the fractionation groove 6a and the sorting groove 6b, the voltage applied to each electrode is switched to sort the desired band into the sorting groove 6b. Finally, the collected bands are collected in the pods 2f to 2m (FIG. 1C: sorting process).

As described above, the microchip electrophoresis apparatus disclosed in Patent Document 1 enables a researcher to sort out a desired DNA without performing complicated operations such as cutting out the gel or extracting from the gel.
JP 2002-310992 A

  However, the microchip electrophoresis apparatus of Patent Document 1 has a problem that the collected sample component is adsorbed to the electrode in the pod.

  That is, in the microchip electrophoresis apparatus of Patent Document 1, since a 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 adsorbed on the electrode in the pod. It will be done. Thus, if the sample component is adsorbed to the electrode, it becomes difficult to take out the collected sample component from the pod using a micropipette or the like. Further, when the electrode is adsorbed and contaminated by the sample component, the ability of the electrode to draw the sample component is reduced.

  An object of the present invention is to provide a microchip and a microchip electrophoresis apparatus that can collect a specific sample component from a sample and collect the collected specific sample component in a state where the sample is not adsorbed to an electrode.

  The microchip of the present invention has a separation channel, a sorting channel intersecting with the separation channel, and a recovery reservoir communicating with the sorting channel, and the separation channel migrates into the sample The collection channel is a microchip that collects the specific component, the collection reservoir is a microchip that collects the specific component, and communicates with the collection reservoir. A recovery electrode reservoir having a recovery electrode for applying a voltage is provided.

  The microchip electrophoresis apparatus of the present invention includes a separation channel having electrodes at both ends, a sorting channel intersecting with the separation channel, a collection reservoir communicating with the sorting channel and having an electrode, and the collection reservoir A microchip having a recovery electrode reservoir having a recovery electrode for applying a voltage to the recovery reservoir, and optically detecting the specific component migrated in the separation channel of the microchip Based on the detection results of the separation channel detector, the recovery reservoir detector optically detecting the specific component recovered in the recovery reservoir of the microchip, and the detection results of the separation channel detector and the recovery reservoir detector A control unit that determines a voltage to be applied to the electrode of the microchip, and a voltage application unit that applies a voltage to the electrode of the microchip based on the determination of the control unit; A.

  According to the present invention, the collected specific sample component can be collected in the collection reservoir without being adsorbed on the electrode. Therefore, according to the present invention, it is possible to quickly collect a specific sample component from the sample and collect the sample component that has been easily collected with a micropipette or the like.

It is a figure for demonstrating operation | movement of the conventional microchip electrophoresis apparatus, (A) is a schematic diagram of the microchip at the time of an introduction process, (B) is a schematic diagram of the microchip at the time of a fractionation process, (C) FIG. 4 is a schematic diagram of a microchip during a sorting process. BRIEF DESCRIPTION OF THE DRAWINGS It is a figure which shows the microchip which concerns on Embodiment 1 of this invention, (A) is a top view of a microchip, (B) is a partially expanded plan view of a microchip, (C) is a partially expanded microchip. It is sectional drawing. It is a schematic diagram which shows the structure of the microchip electrophoresis apparatus of this invention. It is an enlarged plan view of the intersection of a separation channel and a sorting channel. It is a flowchart for demonstrating operation | movement of the microchip electrophoresis apparatus which concerns on Embodiment 1 of this invention. It is a schematic diagram for demonstrating the operation | movement which collect | recovers the sample component of the microchip electrophoresis apparatus which concerns on Embodiment 1 of this invention. It is a top view of the microchip which concerns on Embodiment 2 of this invention. It is a flowchart for demonstrating operation | movement of the microchip electrophoresis apparatus which concerns on Embodiment 2 of this invention. It is a schematic diagram for demonstrating the operation | movement which collect | recovers the sample component of the microchip electrophoresis apparatus which concerns on Embodiment 2 of this invention. It is the photograph of the microchip used in the Example, (A) is a photograph which shows the intersection of an introduction flow path and a separation flow path, (B) is a photograph which shows the intersection of a separation flow path and a preparatory flow path. . It is a photograph which shows the intersection of the introduction flow path and the separation flow path at the time of introduction processing. It is a fluorescence photograph which shows the intersection of a separation channel and a fractionation channel, (A)-(C) is a fluorescence photograph at the time of blocking processing, (D)-(F) is a fluorescence photograph at the time of fractionation processing, ( G) to (I) are fluorescent photographs during the recovery process. It is a fluorescence photograph which shows the intersection of the separation flow path at the time of not performing a blocking process, and a preparatory flow path. It is a chromatogram of fluorescence intensity in the vicinity of the intersection of the separation channel and the sorting channel. It is a fluorescence photograph of the area around the nanochannel in the collection reservoir when a voltage is applied to the collection electrode reservoir.

1. Microchip of the present invention A microchip of the present invention includes a separation channel provided with electrodes at both ends, a sorting channel crossing the separation channel, a collection reservoir communicating with the sorting channel and having an electrode, The microchip further comprises a recovery electrode reservoir in communication with the recovery reservoir and having a recovery electrode.

  The separation channel is a channel that is provided with electrodes at both ends and separates a specific sample component from the sample by electrophoresis of the introduced 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. When the separation channel detection unit described later detects fluorescence, a fluorescent dye may be added to the carrier or buffer solution. The thickness (width × depth) of the separation channel may be appropriately set 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 100 μm, and more preferably in the range of 70 to 100 μm. 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.

  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 inside, and may further be filled with a carrier. The thickness of the sorting channel is not particularly limited, but is preferably thinner 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 μm, and more preferably in the range of 25 to 60 μm. 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.

  The collection reservoir has a pod filled with a buffer solution inside and an electrode for applying a voltage to the sorting channel, and collects a specific sample component sorted in the sorting channel into the pod. . The size of the pod is not particularly limited as long as the sample component can be collected, but a diameter of about 500 μm to 4 mm is preferable, and a depth of about 1 mm is preferable. The material of the electrode is not particularly limited as long as a voltage can be applied to the sorting flow path. However, a metal that is not easily rusted such as gold, platinum, copper, and aluminum is preferable, and gold or platinum is particularly preferable.

  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 but not sample components. The material of the nanochannel is not particularly limited as long as it allows water molecules to pass but does not allow sample components to pass through. For example, a porous material such as porous glass made of sodium silicate 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 interval 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 may be appropriately adjusted according to the material of the nanochannel. That's fine. For example, when sodium silicate porous glass is used for the nanochannel, the length of the nanochannel may be about 5 to 10 μm, and the interval between the collection reservoir and the collection electrode reservoir is about 0.5 to 1 mm. If it is.

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

2. Microchip Electrophoresis Device of the Present Invention The microchip electrophoresis device of the present invention includes the microchip of the present invention, a separation channel 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 near the intersection of the separation channel and the sorting channel of the microchip. At this time, the separation channel detection unit detects not only the sample component migrated in the separation channel, but also the sample component migrated in the sorting channel near the intersection of the separation channel and the sorting channel. It is preferable to do. The method for optically detecting the sample component by the separation flow path detection unit is not particularly limited, and 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.

  The collection reservoir detection unit optically detects the 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 detection unit is not particularly limited, and may be the same method as the separation channel detection unit. The detection result is output to the control unit.

  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 having a polarity opposite to the charge of the sample component to 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 when the desired sample component is migrated in the sorting channel, the control unit Is determined to apply a voltage of the opposite polarity 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 migrating sample components. Whether or not 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 a preliminary experiment has grasped the order in which each sample component in the sample reaches the intersection of the separation channel and the sorting channel, the user can sort out the sample component that reaches the intersection second. ”May be set in the control unit.

  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 include, for example, a power supply and a changeover switch circuit group having a relay switch circuit.

  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.

  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 removes the sample introduced into the separation channel from the introduction point to the separation channel and the sorting channel. Run to the intersection. 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 sieving effect (separation process).

  Next, when the separation channel detection unit detects that a desired sample component has reached the intersection of the separation channel and the sorting channel, the voltage application unit determines whether the microchip is separated based on the determination of the control unit. A voltage is applied to the intake channel. As a result, the desired sample component is taken into the sorting channel (sorting process) and collected in the collecting reservoir (collecting process). Even if the separation channel detection unit detects that an undesired sample component has reached the intersection of the separation channel and the sorting channel, the voltage application unit can determine whether or not the sorting channel is based on the determination of the control unit. Since no voltage is applied to, separation processing is continued without performing preparative processing. Therefore, undesired sample components are further migrated toward the electrode downstream of the separation channel without being taken into the sorting channel.

  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. 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. Since the sample component is prevented from migrating by the nanochannel between the collection reservoir and the collection electrode reservoir, it finally remains in the region around the nanochannel in the collection reservoir.

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

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.

  The sample reservoir includes a pod into which a sample is injected and an electrode that applies a voltage to the sorting channel. The size of the pod is not particularly limited as long as a desired amount of sample components can be injected, but a diameter of about 500 μm to 4 mm is preferable, and a depth of about 1 mm 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.

  The introduction channel is provided with electrodes (one electrode in the sample reservoir) at both ends, and a flow for introducing the sample injected into the sample reservoir into the separation channel from the intersection of the introduction channel and the separation channel. Road. The introduction channel is preferably filled with a buffer solution inside, and may further be filled with a carrier. The thickness (width × depth) of the introduction channel is not particularly limited, but may be, for example, about 90 μm width × 25 μm depth. The length of the introduction flow path is not particularly limited, but for example, the length from the communication point with the sample reservoir to the intersection with the separation flow path may be about 5 mm.

  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.

  First, a sample containing a desired sample component is injected into the sample reservoir. Next, the voltage application unit applies a voltage to the introduction channel to cause the sample to migrate from the sample reservoir toward the intersection of the introduction channel and the separation channel. After the sample reaches the intersection between the introduction channel and the separation channel, the voltage application unit applies a voltage to the separation channel for a short time to introduce a part of the sample into the separation channel.

  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.

4). Block Electrode The microchip of the present invention preferably further has a block electrode.

  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.

  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 Is determined to be applied. A voltage application part applies a voltage to a block electrode based on determination of a control part (blocking process).

  By using the block electrode, it is possible to prevent sample components other than the specific sample component from entering the sorting channel. Therefore, it is possible to prevent the sorting channel from being contaminated by sample components other than the specific sample component.

  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 each other in a cross shape), one block electrode has a voltage across the sorting channel on one side. And the other block electrode applies a voltage to the sorting channel on the other side, thereby preventing the sorting channels on both sides from being contaminated by sample components other than a specific sample component. it can.

  Further, 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 μm, and more preferably in the range of 25 to 60 μm.

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

(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.

  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, a sample discharge reservoir 112, It has a buffer reservoir 114, a buffer discharge reservoir 116, a recovery reservoir 118, a recovery electrode reservoir 120, and two block electrode reservoirs 122a and 122b. The size of the microchip 100 may be, for example, about 100 mm (horizontal direction in the figure) × 30 mm (vertical direction in the figure).

  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 channel 102 may be a channel having a width of 90 μm, a depth of 25 μm, and a length of 10 mm, and intersecting the separation channel 104 at an intermediate point thereof.

  The separation channel 104 intersects the introduction channel 102 and the sorting channel 106 so as to communicate with each other, and is a channel provided with a buffer reservoir 114 and a buffer discharge reservoir 116 at both ends thereof. For example, the separation channel 104 is a channel having a width of 90 μm, a depth of 25 μm, and a length of 85 mm so as to intersect the introduction channel 102 at a point 5 mm from the buffer reservoir 114 and 15 mm from the buffer discharge reservoir 116. What is necessary is just to make it cross | intersect the sorting flow path 106 in the point of.

  The sorting channel 106 intersects with the separation channel 104 and is a channel provided with a recovery reservoir 118 at one end. The sorting channel 106 also communicates with the block channels 108a and 108b. 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 μm, a depth of 25 μm, and a length of 10 mm, and the separation channel 104 at an intermediate point thereof. Should be crossed with. Further, a portion of the sorting channel 106 that is parallel to the separation channel 104 (see FIG. 2A) may be a channel having a width of 90 μm, a depth of 25 μm, and a length of 5 mm, for example.

  The block flow paths 108a and 108b are flow paths that communicate with the sorting flow path 106 at one end, and are provided with block electrode reservoirs 122a and 122b at the remaining end. For example, the block channels 108a and 108b may be channels having a width of 90 μm, a depth of 25 μm, and a length of 20 mm.

  The sample reservoir 110, the sample discharge reservoir 112, the buffer reservoir 114, the buffer discharge reservoir 116, the recovery reservoir 118, the recovery electrode reservoir 120, and the block electrode reservoirs 122a and 122b are each a buffer solution or a sample (sample). ) And the like, and electrodes (not shown) for applying a voltage to each flow path. 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 this specification, applying a voltage to an 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 cross-sectional view of the microchip 100 showing the collection reservoir 118 and the collection electrode reservoir 120. For convenience of explanation, the electrodes are not shown.

  2B and 2C, the recovery electrode reservoir 120 is formed adjacent to the recovery reservoir 118 and communicates with the recovery reservoir 118 via the nanochannel 124 that allows water molecules to pass but does not allow sample components to pass. is doing.

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

  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, the control unit 230, and the voltage application unit 240 shown in FIG. .

  The separation channel detection unit 210 detects fluorescence emitted from the sample component in the vicinity of the intersection of 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 shows the buffer reservoir 114 side, the right side shows the buffer discharge reservoir 116 side, the upper side shows the block electrode reservoir 122a and the recovery reservoir 118 side, and the lower side shows 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, thereby separating the sample component (point a) that has migrated through the separation channel 104 into the sorting channel. The sample component (point b) that has not been detected and the sample component (point c) separated in the sorting channel 106 can be distinguished and detected. The detection result is output to the control unit 230.

  For example, as shown in FIG. 3, the separation channel detection unit 210 guides the excitation light in the direction of the sample component and emits it from the sample component, the mercury lamp 211 as an excitation light source, the excitation filter 212 for separating the excitation light What is necessary is just to make it the structure provided with the dichroic mirror 213 which guide | induces the emitted fluorescence to the CCD camera 215 direction, the absorption filter 214 which isolate | separates fluorescence, and the CCD camera 215 which detects fluorescence. The method for emitting fluorescence from the sample component may be appropriately selected according to the sample component. For example, when 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 component in the collection reservoir 118 of the microchip 100. The recovery reservoir detection unit 220 may have the same configuration as the separation channel detection unit 210, for example. The detection result is output to the control unit 230.

  The controller 230 determines a voltage to be applied to the electrodes in each reservoir of the microchip 100. At this time, the control unit 230 should determine the position of each sample component in the vicinity of the intersection of the separation channel 104 and the sorting channel 106 from the detection result of the separation channel detection unit 210, and should the sample component be sorted? The voltage applied to each electrode of the microchip 100 is determined according to 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 depends on whether or not the sample component is collected in the collection reservoir 118. Thus, the voltage applied to each electrode of the microchip 100 is determined. For example, when the sample component is DNA (negatively charged), a positive voltage may be applied to the downstream electrode of the flow path for migrating the sample component, and the upstream electrode may be grounded.

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

  Next, referring to 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. explain.

  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 to the sample discharge reservoir 112 by applying a voltage to the introduction channel 102, and the sample reaches the intersection of the introduction channel 102 and the separation channel 104. After that, 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, the sample reservoir 110 is grounded and a positive voltage is applied to the sample discharge reservoir 112 in order to migrate the sample from the sample reservoir 110 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, the sample discharge reservoir 112, and the buffer A positive voltage may be applied to the discharge reservoir 116 for 4 seconds.

  Next, in step S1200, a voltage is applied to the separation channel 104 to cause the sample to migrate from the intersection of the introduction channel 102 and the separation channel 104 toward the buffer discharge reservoir 116. At this time, the sample is separated into each sample component by the molecular sieving effect (separation process). For example, when the sample component is DNA, as shown in FIG. 6A, the buffer reservoir 114 is grounded (indicated by “−” in the figure), and a positive voltage may be applied to the buffer discharge reservoir 116 (in the figure “ "+")

  Next, in step S1300, when the separation channel detection unit 210 detects that the sample component separated at the intersection of the separation channel 104 and the sorting channel 106 has arrived (see FIG. 6A), the control unit 230. It is then determined whether this sample component is a desired sample component. When this sample component is not a desired sample component (S1300: NO), the process proceeds to step S1400. On the other hand, when this sample component is a desired sample component (S1300: YES), the process proceeds to step S1500.

  In step S1400, undesired sample components are migrated toward the buffer discharge reservoir 116. At this time, by applying a voltage to the block electrode reservoirs 122a and 122b (block electrodes), the sample component is prevented from entering the sorting channel 106 (blocking process). For example, when the sample component is DNA, when 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 may be grounded (see FIG. 4). (See FIG. 6B). When this sample component completely passes through the intersection of the separation channel 104 and the sorting 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 (see FIG. 6C).

  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 S1600, the collected sample components are recovered in the recovery reservoir 118 (recovery process). For example, when 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 are collected. Can be recovered in the recovery reservoir 118 (see FIG. 6F).

  Next, in step S1700, when the recovery reservoir detector 220 detects that the sample component separated into the recovery reservoir 118 has been recovered, the sample component is applied by applying a voltage to the recovery electrode reservoir 120. The flow is terminated by moving away from the electrode in the collection reservoir 118. For example, when 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. That's fine. By doing so, the sample component adsorbed on the electrode in the collection reservoir 118 is separated from the electrode and further migrates toward the collection electrode reservoir 120. Since the sample component is prevented from migration by the nanochannel 124, the sample component finally remains in the region around the nanochannel 124 in the collection reservoir 118.

  Thus, according to the present embodiment, the collected sample component can be prevented from adsorbing to the electrode in the collection reservoir, and the collected sample component can be collected in the nanochannel peripheral region in the collection reservoir. The sample components in the collection reservoir can be easily taken out using a micropipette or the like.

  In addition, according to the present embodiment, since the block electrode applies a voltage to the sorting channel, it is possible to prevent an undesired band from entering the sorting channel, so that the desired sample component is not contaminated. Can be collected and collected.

  In addition, 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, so that the user performs complicated work. Therefore, the sample components can be accurately sorted by automatic control.

(Embodiment 2)
In the first embodiment, an example in which the recovery reservoir is one 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, a sample discharge reservoir 112, and a buffer. It has a reservoir 114, a buffer discharge reservoir 116, a plurality of collection reservoirs 304a to 304d, a plurality of collection 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 flow path 302 is a flow path that intersects with the separation flow path 104 and branches into a plurality on both sides of the separation flow path 104. Recovery reservoirs 304 a to 304 d are provided at the branch ends of the sorting flow path 302, respectively. For convenience of explanation, the sorting flow path 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 μm, a depth of 25 μm, and a length of 10 mm, and an intermediate point (1/2 point). ) Crossing the separation flow path 104 and branching at both ends, 1/4 points and 3/4 points. The branch channels 302a and 302d are, for example, channels having a width of 90 μm, a depth of 25 μm, and a length of 5 mm, and the branch channels 302b and 302c are channels of, for example, a width of 90 μm, a depth of 25 μm, and a length of 3 mm And it is sufficient.

  The microchip electrophoresis apparatus 200 according to the second embodiment includes the microchip 300 shown in FIG. 7, a separation channel detection unit 210, a plurality of recovery reservoir detection units 220a to 220d, a control unit 230, and a voltage application unit 240. . Components other than the microchip 300 are the same as those in the first embodiment.

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

  In step S2000, when the separation channel detection unit 210 optically detects the arrival of the sample component separated at the intersection of the separation channel 104 and the sorting channel 302 (see FIG. 9A), the control unit 230. It is then determined whether this sample component is a desired sample component. In the second embodiment, the separation channel detection unit 210 detects fluorescence at four points a to d near the intersection between the separation channel and the sorting channel shown in FIG. If this sample component is not the desired sample component (S2000: NO), the process proceeds to step S1400. On the other hand, when the sample component is a desired sample component (S2000: YES), the process proceeds to step S2100.

  In step S2100, a desired sample component is taken into the sorting channel 302 by applying a voltage to the sorting channel 106 (sorting process). For example, in order to collect the desired DNA in the collection reservoir 304a, as shown in FIG. 9D, the buffer discharge reservoir 116 may be opened and a positive voltage may be applied to the collection reservoir 304a.

  Next, in step S2200, the collected sample components are collected in any of the collection reservoirs 304a to 304d (collection process). At this time, the plurality of recovery reservoirs are used from a reservoir 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 reservoir 304a, the collection reservoir 304d, the collection reservoir 304b, and the collection reservoir 304c. For example, in order to recover the desired DNA in the recovery reservoir 304a, the buffer reservoir 114 is opened, the block electrode reservoirs 122a and 122b are grounded, and a positive voltage is applied to the recovery 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 channel 302a, the branch channel 302b is cleaned by applying a voltage to the branch channel 302b located inside the branch channel 302a (cleaning process). . 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. As a result, 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 recovery reservoir detector 220a detects that the sample components separated in the recovery reservoir 304a are recovered, the voltage is applied to the recovery electrode reservoir 306a for a predetermined time. The sample component is separated from the electrode in the collection reservoir 304a. For example, when the collected sample component is DNA, as shown in FIG. 9H, the block electrode reservoirs 122a and 122b are grounded, the collection reservoir 304a is opened, and a positive voltage is applied to the collection electrode reservoir 306a. That's fine. 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 migration by the nanochannel, the sample component finally remains in the region around the nanochannel in the collection reservoir 304a.

  Next, in step S2400, the control unit 230 determines whether to end this flow. For example, when all the collection reservoirs 304a to 304d collect the sample components, when all the desired sample components are collected to the collection reservoirs 304a to 304d, or all the sample components are separated from the separation channel 104 and the sorting channel. When the intersection with 302 is passed, 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, when it is determined that this flow is to be ended (S2400: YES), this flow is ended.

  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.

  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.

  In addition, according to the present embodiment, since the electrodes are arranged in line symmetry with respect to the separation channel, it is possible to avoid the complexity of the structure of the electric circuit and the voltage application switching operation.

  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 may be changed. .

  Hereinafter, the present invention will be further described with reference to examples. It should be noted that the scope of the present invention is not construed as being limited by this example.

  In this example, an example in which 20 bp DNA (specific sample component) is fractionated from a 10 bp DNA ladder (sample) using the microchip electrophoresis apparatus including the microchip of the first embodiment is shown.

  As the microchip used in this example, a glass chip having a flow path shown in FIG. 2A was used. The introduction channel and the separation channel were 90 μm wide × 20 μm deep. On the other hand, the sorting flow path has a width of 50 μm and a depth of 20 μm in order to improve sorting resolution. FIG. 10A shows a micrograph of the intersection between the introduction channel and the separation channel, and FIG. 10B shows a micrograph of the intersection between 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. In FIG. 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, intersected with the introduction channel at a point 5 mm from the buffer reservoir, and intersected with the sorting channel at a point 15 mm from the buffer discharge reservoir. A portion of the sorting channel in the direction perpendicular to the separation channel was 10 mm in length, and intersected with the separation channel at an intermediate point. The part parallel to the separation channel in the sorting channel was 10 mm long. The block channel was 20 mm long.

  In addition, the microchip used in this example was manufactured by laminating a glass plate with a flow channel groove and a glass plate with holes for reservoirs (pods), using sodium silicate as an adhesive layer. Specifically, first, using a general photolithography technique and a wet etching technique, a glass plate with a flow channel groove and a glass plate with a hole for a reservoir (pod) were produced. Next, a thin film of sodium silicate was formed by spin-coating a 0.1 M sodium silicate aqueous solution on one glass plate. The other glass plate was placed on a thin film of sodium silicate and heated at 200 ° C. By doing so, sodium silicate became porous glass, and the two glass plates were bonded to each other. Further, the formed porous glass of sodium silicate was made to function 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 μm.

  A polydimethylacrylamide gel containing a DNA-binding fluorescent dye cyber green and a buffer solution were previously introduced into the channel.

  The separation channel detection unit and the recovery reservoir detection unit are an inverted system microscope (IX70: Olympus) and a CCD camera including a mercury lamp, an excitation filter (460 to 490 nm), a dichroic mirror (505 nm), an absorption filter (510 to 550 nm). (ORCA-EG: Hamamatsu Photonics). The separation channel detection unit was set to monitor changes in fluorescence intensity at points A to C shown in FIG. 10B.

  The voltage switching by the control unit and the voltage application unit was set as follows. In the separation process, the buffer reservoir was grounded, a voltage of +1300 V was applied to the buffer discharge reservoir, and the other reservoirs were opened (see FIG. 6A). The blocking process was set so that the two block electrode reservoirs were grounded when the fluorescence intensity at point B rose above a predetermined threshold (see FIG. 6B). Further, the two block electrode reservoirs were set to return to open when the fluorescence intensity at point B dropped below a predetermined threshold (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 FIG. 6D). The collection process was set such that 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 (see FIGS. 9E and 9F). Further, 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).

(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. Thereby, 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, a voltage of +300 V is applied to the sample discharge reservoir, and a voltage of +1300 V is simultaneously applied to the buffer discharge reservoir for 4 seconds. A part of was introduced into the separation channel.

  FIG. 11 is a fluorescent photograph in the vicinity of 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 (10 bp DNA ladder).

(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). Thereby, the introduced sample started migration toward the buffer discharge reservoir. The sample was separated into a plurality of bands (10 bp DNA, 20 bp DNA,... DNA) by running in a polydimethylacrylamide gel.

(Blocking process)
The plurality of bands continued to migrate toward the buffer discharge reservoir, and reached the intersection of the separation channel and the sorting channel in order from the smallest number of bases. 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 the point B rises above a predetermined threshold, the two block electrode reservoirs were grounded. Thereby, this band (10 bp DNA) migrated toward the buffer discharge reservoir without entering the sorting channel (see FIG. 6B). When the fluorescence intensity at the point B dropped below a predetermined threshold, the two block electrode reservoirs were returned to the open state, and the electrophoresis continued (see FIG. 6C).

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

  FIG. 13 is a fluorescent photograph in the vicinity of the intersection of the separation channel and the sorting channel when the blocking process is not performed as a comparative example. The direction of each flow path is the same as the photograph in FIG. From this photograph, it can be seen that undesired bands (100 bp DNA and 110 bp DNA) would enter the sorting channel without blocking treatment.

(Preparation processing)
Since the next reached band (20 bp DNA) is the desired DNA, a sorting process and a collecting process 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 recovery reservoir (see FIG. 6D). Thereby, the desired band (20 bp DNA) was fractionated into the sorting channel.

  12D to 12F are fluorescent photographs near the intersection of the separation channel and the sorting channel during the sorting process. 12D is a photograph when it is detected that the fluorescence intensity at the point A has risen above a predetermined threshold, FIG. 12E is a photograph when the sorting process is started, and FIG. 12F is a photograph when the sorting process is finished. . The direction of each flow path is the same as the photograph of FIG. 12A. From these photographs, it can be seen that the desired band (20 bp DNA) was separated by performing the sorting process.

(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 dropped 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 recovery reservoir and was finally recovered in the recovery reservoir (see FIGS. 6E and 6F).

  12G to 12I are fluorescent photographs in the vicinity of 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. 12I is a photograph when the collection process is finished. The direction of each flow path is the same as the photograph of FIG. 12A. From these photographs, it can be seen that the desired band was recovered as intended by performing the recovery 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. Further, the arrow of the chromatogram at the point A (upper stage) indicates the start time of the desired band sorting process, and the arrow of the chromatogram at the point C (middle stage) indicates the start time of the desired band recovery process. FIG. 14 shows that an undesired band (first band) is detected at point A (upper stage) and then detected at point B (lower stage), so that this band is not sorted. On the other hand, since a desired band (second band) is detected at point A (upper stage) and then detected at point C (middle stage), it can be seen that only this band is sorted.

(Voltage applied to collection electrode reservoir)
After the recovery process, a process for separating the recovered sample component from the electrode in the recovery reservoir was performed. That is, when the fluorescence intensity in the recovery reservoir rose above a predetermined threshold, the potential of the recovery reservoir was opened, and a voltage of +1300 V was applied to the recovery electrode reservoir. As a result, the sample component (20 bp DNA) in the collection reservoir gathered around the nanochannel in the collection reservoir. (See FIG. 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 the one that appears white is the sample 400 (20 bp DNA ladder) fluorescently stained with Cyber Green. is there. Thus, it can be seen that by applying a voltage to the collection electrode reservoir, the sample component (20 bp DNA) in the collection reservoir is separated from the electrode in the collection reservoir and collects in the nanochannel peripheral region.

  This application claims the priority based on Japanese Patent Application No. 2006-273377 filed on Oct. 4, 2006. The contents described in the application specification and the drawings are all incorporated herein.

  Since the microchip and the microchip electrophoresis apparatus according to the present invention can sort sample components such as DNA without a complicated work by a researcher, analysis of biological materials related to all bio industries, etc. Useful for.

Claims (10)

A separation channel, a sorting channel intersecting with the separation channel, and a collection reservoir in communication with the sorting channel and having an electrode;
The separation channel separates a specific component contained in a sample to be migrated therein, the sorting channel separates the specific component, and an electrode of the recovery reservoir causes the specific component to be electrophoresed. A microchip to be collected in a collection reservoir,
A microchip having a recovery electrode reservoir having a recovery electrode that communicates with the recovery reservoir and applies a voltage to the recovery reservoir.   The microchip according to claim 1, wherein the collection electrode reservoir communicates with the collection reservoir via a porous substance.   The microchip according to claim 2, wherein the porous material is sodium silicate.   The microchip according to claim 1, wherein the sorting channel is narrower than the separation channel. A sample reservoir;
An introduction channel communicating with the sample reservoir and intersecting the separation channel;
The microchip according to claim 1, further comprising:   The microchip according to claim 1, further comprising a block electrode that applies a voltage to the sorting flow path to prevent components other than the specific component from entering the sorting flow path.   The microchip according to claim 6, wherein the number of the block electrodes is two or more.   The microchip according to claim 7, wherein the block electrode is disposed on both lateral sides of the separation channel. Separation flow path having electrodes at both ends, preparative flow path intersecting with the separation flow path, communicating with the preparative flow path, collecting reservoir having electrodes, and communicating with the collecting reservoir, and applying a voltage to the collecting reservoir A recovery electrode reservoir having a recovery electrode to be applied;
The separation channel separates a specific component contained in a sample to be migrated therein, the sorting channel separates the specific component, and an electrode of the recovery reservoir causes the specific component to be electrophoresed. A microchip to be collected in a collection reservoir;
A separation channel detector for optically detecting a specific component migrated in the separation channel of the microchip;
A recovery reservoir detector that optically detects the specific component recovered in the recovery reservoir of the microchip;
A control unit for determining 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. Separation flow path having electrodes at both ends, separation flow path intersecting with the separation flow path, communicating with the separation flow path, collecting reservoir having electrodes, communicating with the collection reservoir, and applying a voltage to the collection reservoir A recovery electrode reservoir having a recovery electrode, and a block electrode that suppresses entry of components other than the specific component into the sorting flow path by applying a voltage to the sorting flow path,
The separation channel separates a specific component contained in a sample to be migrated therein, the sorting channel separates the specific component, and an electrode of the recovery reservoir causes the specific component to be electrophoresed. A microchip to be collected in a collection reservoir;
A separation channel detection unit for optically detecting the specific component migrated in the separation channel of the microchip;
A recovery reservoir detector that optically detects the specific component recovered in the recovery reservoir of the microchip;
A control unit for determining 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.