JP2012098309A - Capillary array unit and capillary electrophoretic apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a capillary array unit constituted so as to simplify mounting work of a capillary array, and a capillary electrophoretic apparatus.SOLUTION: A capillary array unit comprises: a capillary array including a capillary equipped with a capillary head and a detection part; a frame for supporting the capillary array; and a load header for holding a cathode end of the capillary. The frame has a separator for separating and holding the capillary. The capillary head, the detection part, and the separator are arranged along one straight line.

Description

  The present invention relates to a capillary array unit and a capillary electrophoresis apparatus using the same.

  Capillary electrophoresis is widely used as a technique for separating and analyzing many biological samples including deoxyribonucleic acid (DNA). One of its technical advantages is the excellent heat dissipation properties resulting from the surface area of the capillary versus the volume ratio. This heat dissipation characteristic realizes high-speed and high-resolution sample separation by electrophoresis using a high voltage.

  Japanese Patent Laid-Open No. 2001-281221 discloses a pump mechanism for filling a capillary with a polymer as a separation medium. Furthermore, a method using a motor stall torque is disclosed as means for generating pressure for polymer filling.

  Japanese Patent Application Laid-Open No. 2001-324473 discloses an electrophoresis apparatus including an oven that can accommodate capillaries having different lengths or numbers depending on the type of analysis and the throughput required by the user. Furthermore, this oven uses a Peltier device as a heat source, and can be set from a temperature below room temperature to a temperature above 50 ° C.

  Japanese Unexamined Patent Application Publication No. 2001-324475 discloses a capillary array that can be replaced by a user. The capillary array has a structure that is held at three locations: a sample introduction end, an optical detection portion, and a polymer solution supply end. The polymer solution supply end of the capillary array is connected to the pump mechanism of the electrophoresis apparatus.

JP 2001-281221 A JP 2001-324473 A JP 2001-324475 A

  In a capillary electrophoresis apparatus, when the type of sample or application changes, the capillary array is replaced accordingly. The user replaces the capillary array. There are two types of capillaries, one with a relatively strong type and one with a relatively flexible type. In the case of a flexible type, the replacement work of the capillaries is relatively easy.

  However, in the case of a rigid type, it is difficult to replace the capillary. The capillaries can be linear or warped due to the gravity of the part being applied. When the capillary is mounted in a predetermined position such as an oven, the capillary must be deformed. Since it is difficult to deform the capillaries, it is difficult to replace the capillaries.

  In addition, when the number of capillaries is large, the rigidity of the capillary array becomes strong and the mounting operation becomes difficult.

  An object of the present invention is to provide a capillary electrophoresis apparatus configured to simplify the mounting operation of the capillary array.

  The present invention relates to a capillary array unit and a capillary electrophoresis apparatus using the same. The capillary array unit includes a capillary array including capillaries provided with a capillary head and a detection unit, a frame that supports the capillary array, and a load header that holds the cathode end of the capillary. The frame has at least one separator that separates and holds the capillaries. As the capillaries pass through the separator, the capillary array is held in a certain shape.

  According to the present invention, the mounting operation of the capillary array is simplified.

It is a figure which shows the basic composition of the capillary electrophoresis apparatus by this invention. It is a figure which shows the basic composition of the capillary array by this invention. It is a figure which shows the mounting part of the load header of the capillary electrophoresis apparatus by this invention. It is a figure which shows the capillary array after mounting the load header of the capillary electrophoresis apparatus by this invention. It is a figure which shows the mounting part of the capillary head of the capillary electrophoresis apparatus by this invention. It is a figure which shows the mounting part of the detection part of the capillary electrophoresis apparatus by this invention.

  FIG. 1 is a schematic diagram showing a basic configuration of a capillary electrophoresis apparatus. The capillary electrophoresis apparatus includes a capillary electrophoresis unit 1 including one or a plurality of capillaries, an optical detection unit 2 that optically detects a sample separated by an electrophoresis medium in a capilla, and a high viscosity that is an electrophoresis medium in the capillary. A polymer injection mechanism 3 for injecting a polymer solution (hereinafter referred to as polymer) is provided.

  The capillary electrophoresis unit 1 includes a capillary array 110, an oven (constant temperature bath) 115, a buffer reservoir (buffer container) 112, and a high voltage power source 114.

  The capillary array 110 includes one or a plurality of capillaries. Here, it is assumed that the capillary array 110 includes 16 or 24 capillaries. The capillary is a quartz pipe, and the jacket is coated with a polyimide resin. There are two types of capillaries: a type with a relatively large diameter and strong rigidity, and a type with a relatively small diameter and flexibility. As an example of a type having high rigidity, there is a capillary whose outer diameter is 320 μm and inner diameter is 50 μm. The thickness of the polyimide coating is 20 μm. Therefore, the outer diameter of the polyimide coating is 360 μm. As an example of a flexible type, there is a capillary having an outer diameter of 125 μm and an inner diameter of 50 μm. The thickness of the polyimide coating is 12.5 μm. Therefore, the outer diameter of the polyimide coating is 150 μm.

  According to this example, the capillary array 110 includes a capillary having a relatively large diameter and strong rigidity. The outer diameter of the capillary is about 0.3 to 0.7 mm, and the inner diameter is about 0.02 to 0.2 mm.

  One end of the capillary array 110 is a capillary head 203 in which capillaries are bundled and bonded. The other end of the capillary array 110 is held by the load header 202. The load header 202 is fixed to the oven 115.

  The load header 202 is provided with a tubular cathode electrode 204. The capillary passes through the cathode electrode 204 and protrudes from the lower end of the cathode electrode 204. Thus, the capillary cathode end 206 is immersed in the buffer solution in the buffer reservoir 112.

  The oven 115 accommodates the capillary array 110 and adjusts the temperature of the capillary array 110. A Peltier element is used as a heat source of the oven 115, and the temperature can be set from a temperature lower than room temperature to a high temperature of 50 ° C. or higher.

  The polymer injection mechanism 3 includes a pump 103 having a plunger, a block 104 having a flow path therein, a polymer bottle 101 for storing the polymer, and a buffer container 107 for storing the buffer solution. The anode electrode 106 is immersed in the buffer solution in the buffer container 107. The inner diameter of the flow path in the block 104 is 0.5 to 2 mm, which is several to several tens of times larger than the inner diameter of the capillary. This is to avoid the occurrence of voltage loss during electrophoresis.

  The block 104 is connected to a pump 103, a capillary head 203, and two pipes 104a and 104b. The pump 103, the capillary head 203, and the two pipes 104a and 104b are connected to each other by a flow path in the block 104. The first pipe 104 a connects between the block 104 and the polymer in the polymer bottle 101. A check valve 102 is provided in the first pipe 104a. The second tube 104 b connects between the block 104 and the buffer solution in the buffer reservoir 107. An electric buffer valve 105 is provided in the second pipe 104b.

  The polymer bottle 101 stores a sufficient amount of polymer necessary for continuous operation. An exhaust valve is provided in the polymer bottle 101 or a sufficient gap is provided in the tube insertion port so that the inside of the polymer bottle does not become negative pressure even if the polymer is sucked from the polymer bottle 101. The polymer bottle 101 is disposed at a position lower than the buffer container 107. This is to prevent the polymer from flowing backward from the polymer bottle 101 to the buffer container 107 due to the pressure difference. Conversely, the backflow of the polymer or buffer solution into the polymer bottle 101 is blocked by the check valve 102. The liquid levels of the buffer liquid in the two buffer reservoirs 112 and 107 are held at the same height.

  When the polymer is injected into the capillaries of the capillary array 110, the electric buffer valve 105 is closed. Thereby, the flow path between the capillary array 110 and the buffer reservoir 107 is closed. By driving the pump 103, the polymer in the polymer bottle 101 is injected into the capillary. When performing electrophoresis, the buffer valve 105 is opened, and the flow path between the capillary array 110 and the buffer reservoir 107 is connected.

  The optical detection unit 2 includes a light source 111 and an optical detector 108. The optical detection unit 2 is disposed in the detection unit 205 provided in the capillary array 110. The detection unit 205 is attached to the detection unit holder 116. The light source 111 generates laser light as excitation light. In the detection unit 205, the capillary coating is removed, and the quartz pipe is exposed. Excitation light from the light source 111 irradiates a detection target to be electrophoresed in the capillary in the detection unit 205. Fluorescence is generated from the detection target. This fluorescence is detected by the optical detector 108.

  An electrophoresis method will be described. Although omitted in FIG. 1, an autosampler for transporting the sample tray and the buffer container 112 is provided. A sample tray is placed on the cathode end 206 of the capillary by an autosampler. First, a sample tray is placed under the cathode end 206 of the capillary, and then the sample tray is raised. The sample tray has a large number of wells, and each well contains a sample containing a test object such as fluorescently labeled DNA. The cathode end 206 of the capillary 201 is immersed in the sample in the well of the sample tray. Next, a high voltage of about several kV is applied between the anode electrode 106 and the cathode electrode 204 by the high voltage power supply 114. A detection target such as fluorescently labeled DNA is introduced into the capillary through the cathode end 206 of the capillary. Thereafter, the cathode end 206 of the capillary is immersed in the buffer container 112 as shown in FIG. The detection target is separated while moving in the capillary. When the fluorescently labeled detection target passes through the detection unit 205, excitation light from the light source 111 is irradiated. The detection target generates fluorescence by the excitation light. This fluorescence is detected by the optical detector 108.

  The operation of the polymer injection mechanism will be described. In the pump 103, the direction in which the plunger is pushed into the chamber is assumed to be normal rotation of the motor, and the direction in which the plunger is drawn is assumed to be reverse of the motor. First, the valve 105 is closed. Next, the motor is reversed. The plunger is drawn, and the polymer in the polymer bottle 101 is sucked into the chamber of the pump 103 via the flow path in the block 104. Next, the motor is rotated forward. The plunger is pushed, and the polymer in the chamber of the pump 103 is pushed into the flow path in the block 104. At this time, the polymer in the chamber of the pump 103 is prevented from flowing back to the polymer bottle 101 by the action of the reverse support valve 102. Therefore, the polymer flows into the capillary via the flow path in the block 104 and flows out from the cathode end 206 of the capillary. Finally, the valve 105 is opened to prepare for electrophoresis.

  An example of the capillary array unit of the present invention will be described with reference to FIGS. 2A and 2B. FIG. 2A shows a front configuration of the capillary array unit of this example, and FIG. 2B shows a side configuration thereof. The capillary array unit of this example includes a capillary array 110 and a frame 300 that holds the capillary array 110. As shown in FIG. 2A, the frame 300 includes a first foot 301, a first support portion 302, a second foot 303, a second support portion 304, and first and second bridges 305 and 306. Have. The frame 300 is configured to hold the capillary array 110 on one plane. The two legs 301 and 303 are fixed to the load header 202. The two support portions 302 and 304 are connected by a connection portion 307. The first bridge 305 bridges the connection point between the first foot 301 and the first support portion 302 and the second foot 303. The second bridge 306 bridges the first support portion 302 and the second foot 303.

  The first support portion 302 is provided with two shafts 311 and 312. A shaft 313 is provided on the second leg 303.

  As shown in FIG. 2B, these axes 311, 312, and 313 extend so as to be orthogonal to the plane that forms the frame 300. Separators 321, 322, and 323 are attached to the shafts 311, 312, and 313. The separator is a film or a plate and has the same number of holes as the number of capillaries. The inner diameter of the hole is slightly larger than the outer diameter of the capillary, for example, about φ1 mm. One capillary passes through each hole. Thus, all capillaries are held by penetrating the separator holes.

  The separator separates the capillaries from each other and prevents the capillaries from being intertwined with each other and from being densely bundled. The number of separators may be increased or decreased depending on the length of the capillary. Usually, the longer the capillary, the greater the number of separators.

  The capillary array 110 has a plurality of capillaries 201. The capillary array 110 of this example has 24 capillaries 201. According to this example, the capillary array 110 includes a capillary having a relatively large diameter and strong rigidity. The outer diameter of the capillary is about 0.3 to 0.7 mm, and the inner diameter is about 0.02 to 0.2 mm.

  One end of the capillary array 110 is a capillary head 203 in which capillaries are bundled and bonded. The other end of the capillary array 110, that is, the cathode end is held by a tubular electrode provided on the load header 202.

  Since the capillary of this example has relatively large rigidity, it has a linear shape unless external force and restraining force are applied. In this example, the capillary array is constrained by the load header 202 and the frame 300. As illustrated, the capillary array has a straight portion 110A and a curved portion 110B. The straight portion 110 </ b> A protrudes from the frame 300 and is not restrained by the frame 300. The linear portion 110A includes a capillary head 203 and a detection unit 205. Regardless of the weight of the capillary head 203 and the detection unit 205, the linear portion 110A has a substantially linear shape.

  The curved portion 110B is restrained by the load header 202 and the separator. The shape of the capillary array is determined by the positions of the load header 202 and the separator when no external force is applied to the capillary array.

  Next, the shape of the capillary array will be described. According to this example, the linear portion 110A of the capillary array is arranged along the first support portion 302 of the frame. The capillary head 203, the detection unit 205, and the separator 321 mounted on the shaft 311 are arranged on a straight line. Thus, according to the capillary array unit of this example, substantially half of the capillary array is arranged in a substantially straight line.

  An external force in the direction indicated by arrow A is applied to the capillary head 203. Then, the linear portion 110 </ b> A moves along the direction indicated by the arrow A. Thereby, the capillary slides in the holes of the separators 321 and 322. Thus, the capillary head 203 moves in the direction indicated by the arrow A.

  FIG. 2A shows the shape of the capillary array of this example when no external force is applied to the capillary array. When the capillary array unit of this example is attached to the electrophoresis apparatus, the capillary array is held in the shape shown in FIG. 2A. That is, even if the capillary array unit of this example is mounted on the electrophoresis apparatus, the capillary array retains the shape shown in FIG. 2A.

  Therefore, the relative positional relationship among the capillary head 203, the detection unit 20, and the load header 202 in the capillary array shown in FIG. 2A is the same as the capillary head 203, the detection unit 205, and the load in the capillary array mounted on the electrophoresis apparatus. The relative position between the mounting positions of the header 202 is equal to the positional relationship.

  According to this example, the position of the separator is selected so that the capillary array has the shape shown in FIG. 2A. Thus, by selecting the position of the separator, it is not necessary to apply an external force to the capillary array when the capillary array 110 is mounted on the electrophoresis apparatus. Therefore, the capillary array can be mounted without being deformed.

  According to this example, as shown in FIG. 2A, the capillary head is disposed below, and the linear portion 110A of the capillary array is disposed in an inclined manner. This will be described. In the capillary electrophoresis apparatus shown in FIG. 1, in order to reduce the amount of polymer used, it is better to shorten the tube 104b connected to the block 104. Shortening the tube 104b not only reduces the amount of polymer used, but also reduces the material cost of the tube 104b. If the tube 104b is shortened, it is necessary to lower the block 104 or raise the buffer reservoir 107. On the other hand, the liquid level of the buffer reservoir 107 and the liquid level of the buffer reservoir 112 need to be the same level. Therefore, the buffer reservoir 107 cannot be raised. Therefore, the block 104 is lowered. That is, the block 104 is arranged as low as possible. When the block 104 is lowered, the mounting position of the capillary head is also lowered. Therefore, as shown in FIG. 2A, half of the capillary array is inclined.

  Next, with reference to FIG. 3, FIG. 4, FIG. 5 and FIG. 6, a procedure for mounting the capillary array according to the present invention will be described. First, the load header 202 is attached to the oven 115. This will be described with reference to FIG. FIG. 3 shows a part of the lower end of the oven 115 and the load header 202. Illustration of the capillary array mounted on the load header is omitted. The load header 202 is provided with a grip 202a. The user grasps the grip 202 a and inserts the load header 202 into the recess of the oven 115. Grooves are provided on both side surfaces of the load header 202, and protrusions are provided on the inside of the recesses of the oven 115. When the load header 202 is inserted into the recess of the oven 115, the groove of the load header 202 and the protrusion of the recess of the oven 115 are engaged.

  FIG. 4 shows a state in which the load header 202 is attached to the oven 115. The shape of the capillary array 110 shown in FIG. 2A is maintained as it is. In this example, the capillary head 203 is disposed in the vicinity of the mounting portion of the block 104 of the polymer injection mechanism 3. In addition, the detection unit 205 is disposed in the vicinity of the mounting unit of the detection unit holder 116.

  Next, the capillary head 203 is mounted on the block 104 of the polymer injection mechanism 3. This will be described with reference to FIG. The ferrule 501 is loaded into the hole of the block 104, and the capillary head 203 is passed through the ferrule 501. Alternatively, the capillary head 203 may be passed through the ferrule 501, and then the capillary head 203 may be inserted into the hole of the block 104 together with the ferrule 501. At this time, as described with reference to FIG. 2, the capillary head 203 can be moved in the front-rear direction with a small force. Next, the push screw 502 is screwed and the ferrule 501 is crushed. Thereby, the space between the capillary head 203 and the block 104 is sealed, and the capillary head 203 is fixed to the block 104.

  As shown in FIG. 4, the capillary head 203 is disposed in the vicinity of the mounting portion of the block 104 of the polymer injection mechanism 3. Therefore, this operation is easy because the capillary head 203 is moved slightly in the front-rear direction.

  Finally, the detection unit 205 is attached to the detection unit holder 116. This will be described with reference to FIG. The detection unit 205 is fitted into a frame provided in the detection unit holder 116. Next, the lid is closed.

  As shown in FIG. 4, the detection unit 205 is disposed in the vicinity of the mounting unit of the detection unit holder 116. Therefore, this operation is easy because the detection unit 205 is slightly moved in the lateral direction.

  As described above, according to this example, it is not necessary to deform the capillary array when the capillary array is attached to the electrophoresis apparatus. Therefore, the mounting operation of the capillary array can be easily performed. Accordingly, the replacement operation of the capillary array is facilitated.

  Although the example of the present invention has been described above, the present invention is not limited to the above-described example, and it is easy for those skilled in the art that various modifications can be made within the scope of the invention described in the claims. Will be understood.

DESCRIPTION OF SYMBOLS 101 ... Polymer bottle, 102 ... Check valve, 103 ... Pump, 104 ... Block, 105 ... Buffer valve, 106 ... Electrode, 107 ... Buffer container, 108 ... Photo detector, 110 ... Capillary array, 111 ... Light source, 112 ... Buffer Container 113 113 Electrode 114 High voltage power source 115 Oven 116 Detection unit holder 201 Capillary 202 Load header 202a Grip 203 Capillary head 204 Electrode 205 Detection unit 205a 205b ... Opening, 300 ... Frame, 301 ... Foot, 302 ... Supporting part, 303 ... Foot, 304 ... Supporting part, 305,306 ... Bridge, 307 ... Connecting part, 311,312,313 ... Shaft, 321,322, 323 ... Separator

Claims (9)

A capillary array including one or more capillaries that can be filled with an electrophoretic medium, and at least one separator, and the capillaries pass through the separator to separate the capillaries and hold them in a fixed shape In a capillary array unit having a frame to be loaded, and a load header holding the cathode end of the capillary,
The capillary array is
A capillary head formed at the end opposite to the cathode end;
A detection unit provided between the cathode end and the capillary head;
Have
The capillary array is
It consists of a linear part protruding from the frame and a remaining curved part held by the frame, the linear part having the capillary head at the tip, and the curved part at the one end A capillary array unit having a cathode end. The capillary array unit according to claim 1,
The frame is
First and second legs fixed to the load header;
First and second support portions respectively connected to the first and second legs;
A connecting portion for connecting the first and second support portions;
A first bridge that bridges between the first and second legs;
A second bridge that bridges between the first and second support portions;
Have
The capillary array unit is configured to hold the capillary array on one plane. A capillary array unit including one or more capillaries that can be filled with an electrophoretic medium, a frame that supports the capillary array, and a load header that holds a cathode end of the capillary, and the capillary array An oven for maintaining the temperature at a constant temperature, an injection mechanism for injecting an electrophoretic medium into the capillary, a light source for irradiating excitation light, and an optical detection unit including a light receiving unit for detecting fluorescence, In a capillary electrophoresis apparatus having
The capillary array is
A capillary head formed at the end opposite to the cathode end;
A detection unit provided between the cathode end and the capillary head;
The capillary head is connected to the injection mechanism,
The frame is
Having at least one separator for separating and holding the capillary;
When the capillary passes through the separator, the capillary array is held in a certain shape,
The capillary array is
It consists of a linear part protruding from the frame and a remaining curved part held by the frame, the linear part having the capillary head at the tip, and the curved part at the one end A capillary electrophoresis apparatus having a cathode end. The capillary electrophoresis apparatus according to claim 3,
The frame is
First and second legs fixed to the load header;
First and second support portions respectively connected to the first and second legs;
A connecting portion for connecting the first and second support portions;
A first bridge that bridges between the first and second legs;
A second bridge that bridges between the first and second support portions;
The capillary electrophoresis apparatus is configured to hold the capillary array on one plane. A capillary array unit including one or more capillaries that can be filled with an electrophoretic medium, a frame that supports the capillary array, and a load header that holds a cathode end of the capillary, and the capillary array An oven for maintaining the temperature at a constant temperature, an injection mechanism for injecting an electrophoretic medium into the capillary, a light source for irradiating excitation light, and an optical detection unit including a light receiving unit for detecting fluorescence, In a capillary electrophoresis apparatus having
The capillary array is
The straight part protrudes from the frame and the remaining curved part held by the frame. The straight part has a capillary head at the tip, and the curved part is at the end of the shadow part. The linear portion further includes a detection unit for irradiating excitation light from the light source;
The capillary head is
The capillary electrophoresis apparatus, wherein the linear portion is inclined so as to be arranged at a position lower than the detection unit. The capillary electrophoresis apparatus according to claim 5, wherein
The frame has at least one separator for separating and holding the capillary;
The capillary electrophoresis apparatus, wherein the capillary head, the detection unit, and the at least one separator are arranged along one straight line. The capillary electrophoresis apparatus according to claim 5, wherein
The relative position between the capillary head and the detection unit after the capillary array unit is mounted on the electrophoresis apparatus is the position of the capillary head and the detection unit before the capillary array unit is mounted on the electrophoresis apparatus. A capillary electrophoresis apparatus characterized by being configured to have the same relative position. The capillary electrophoresis apparatus according to claim 6, wherein
The shape of the capillary array after mounting the capillary array unit on the electrophoresis apparatus is configured to be the same as the shape of the capillary array before mounting the capillary array unit on the electrophoresis apparatus. A capillary electrophoresis device characterized. The capillary electrophoresis apparatus according to claim 5, wherein
The frame is
First and second legs fixed to the load header;
First and second support portions respectively connected to the first and second legs;
A connecting portion for connecting the first and second support portions;
A first bridge that bridges between the first and second legs;
A second bridge that bridges between the first and second support portions;
The capillary electrophoresis apparatus is configured to hold the capillary array on one plane.

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