JP6610454B2 - Microchip electrophoresis device - Google Patents

Description

  The present invention relates to a microchip electrophoresis apparatus.

  In microchip electrophoresis, a microchip for electrophoresis (hereinafter also referred to as a chip) having a flow path including a separation flow path inside a plate-like member is used. Samples such as DNA, RNA, or protein introduced into the separation channel of the chip are separated and detected by applying voltage across the separation channel and electrophoresis in the separation channel. Is done.

  The microchip is formed, for example, by using a pair of plate-like members and bonding them with the flow path inside. Microchip electrophoresis is a technique that can separate and detect nucleic acids, proteins, and the like at high speed. In order to keep running costs as low as possible, it is preferable to process multiple samples at a time or reuse the chip repeatedly. In order to reuse the chip repeatedly without sacrificing convenience, preparation for analysis of the chip (cleaning of the chip, filling of the separation buffer), sample dispensing, electrophoresis, post-analysis post-processing (for sample and separation buffer) It is necessary to repeat suction removal and chip cleaning in a fully automatic manner, and a complicated mechanism is required (see Patent Document 1).

Japanese Patent No. 4375031

  An object of the present invention is to simplify the configuration of a microchip electrophoresis apparatus.

  An embodiment of a microchip electrophoresis apparatus according to the present invention includes a flow path including at least a separation flow path for separating a sample by electrophoresis, and a reservoir opened at each end of the flow path. A chip holder for holding a microchip, a probe support for supporting a dispensing probe for dispensing liquid into the reservoir, and a nozzle support for supporting a suction nozzle for sucking the liquid in the reservoir And a moving mechanism that moves one of the probe support and the nozzle support, and the probe support and the nozzle support are detachably connected to each other by the moving mechanism. They are moved at the same time.

  In an embodiment of a microchip electrophoresis apparatus according to the present invention, a nozzle support and a probe support are detachably connected, and both supports are connected by a moving mechanism that moves one of the supports. It is not necessary to provide each drive unit for moving each support, and both supports are moved by a moving mechanism for moving one of the supports. Can be made. Thereby, a structure can be simplified.

It is a block diagram which shows roughly the part regarding the control part in one Embodiment of a microchip electrophoresis apparatus. It is a typical perspective view of the embodiment. It is a typical top view of the embodiment. It is a typical side view including a partial cross section for demonstrating the operation | movement of the embodiment. It is a figure which shows an example of a microchip, (A) and (B) are top views which show the transparent plate-shaped member which comprises a microchip, (C) is a front view of a microchip. It is a top view which shows a specific example of the microchip. It is a schematic top view which shows the chip | tip holding | maintenance part of the state in which the chip | tip is mounted. It is a schematic plan view which shows a cover. It is a schematic perspective view for demonstrating the operation | movement of the embodiment to process order. FIG. 6 is a schematic perspective view for explaining subsequent operations of the embodiment in order of processes. FIG. 22 is a schematic perspective view for explaining a subsequent operation of the embodiment in order of processes. FIG. 22 is a schematic perspective view for explaining a subsequent operation of the embodiment in order of processes. FIG. 22 is a schematic perspective view for explaining a subsequent operation of the embodiment in order of processes. It is the side view which showed the connection state of a pressurized liquid feeding part and a chip | tip typically in the partial cross section.

  In an embodiment of the microchip electrophoresis apparatus of the present invention, either the probe support or the nozzle support includes a pin provided so as to extend vertically downward, and both the supports are the pins. You may come to be connected via.

  As a preferred aspect of the above embodiment, the pin is a pre-punch pin provided on the probe support, and the nozzle support is provided with a hole into which a tip of the pre-punch pin is inserted. The aspect comprised so that both the said support bodies may be connected by being inserted in the hole can be mentioned. The sample container installed in the microchip electrophoresis apparatus may be covered with a protective sheet made of, for example, aluminum or resin. Therefore, the microchip electrophoresis apparatus may be provided with a pre-punch pin for making a hole in the protective sheet. In this embodiment, since the probe support and the nozzle support are connected using the pre-punch pins supported by the probe support, the probe support and the nozzle support can be connected with a simple configuration. The method for connecting the probe support and the nozzle support is not limited to the method using a pre-punch pin. For example, the probe support and the nozzle support fitted to the protrusion or recess provided on the probe support are provided. Any method may be used, such as using a concave portion or a protruding portion.

  In the embodiment of the microchip electrophoresis apparatus of the present invention, for example, the microchip electrophoresis apparatus further includes a nozzle guide mechanism that guides the moving direction of the nozzle support, and the probe moving mechanism is a triaxial direction that is orthogonal to each other and includes a vertical direction. A probe support mechanism for movably supporting the probe support, and a drive device for moving the probe support in the three-axis direction, wherein the nozzle guide mechanism includes the vertical direction of the three-axis directions. A nozzle support mechanism that movably supports the nozzle support in the biaxial direction including Thereby, a probe moving mechanism and a nozzle guide mechanism can be realized with a simple configuration. Here, the vertical direction includes not only a strictly vertical direction but also a roughly vertical direction. The configurations of the probe moving mechanism and the nozzle guide mechanism are not limited to these. For example, the probe moving mechanism may be a mechanism that can freely move the probe support in three dimensions, such as a vertical or horizontal articulated robot. Further, the nozzle guide mechanism is not limited to the one that guides the nozzle support along a straight line, and may be one that guides along a curve.

Hereinafter, an embodiment of a microchip electrophoresis apparatus will be described with reference to the drawings.
FIG. 1 is a block diagram schematically showing a portion related to a control unit in one embodiment of a microchip electrophoresis apparatus. FIG. 2 is a schematic perspective view of this embodiment. FIG. 3 is a schematic plan view of this embodiment. FIG. 4 is a schematic side view including a partial cross-section for explaining the operation of this embodiment.

  The microchip electrophoresis apparatus 1 roughly includes a dispensing probe mechanism 6 having a dispensing probe section 2, a liquid level detection section 3, and a syringe pump section 4, a chip holding section 7, a pressurized liquid feeding section 16, and a suction pump section 18. A suction nozzle mechanism 19, a voltage application unit 24, a detection unit 31, and a control unit 38.

  For example, one chip 5 (microchip) is held in the chip holding unit 7. A cover 26 is disposed facing the chip 5. The cover 26 is detachably attached to the chip holding part 7 with screws 27. The chip 5 will be described later.

  The dispensing probe mechanism 6 performs dispensing of separation buffer, reagent, and sample to the reservoir of the chip 5, supply of cleaning liquid, detection of remaining amount of separation buffer and reagent, and the like. The dispensing probe unit 2 of the dispensing probe mechanism 6 includes a dispensing probe 8, a pre-punch pin 11, a probe support 12, and a probe moving mechanism 100.

  The dispensing probe 8 is supported by the probe support 12. The pre-punch pin 11 is supported by the probe support 12 so as to be detachable. A mechanism for supporting the pre-punch pin 11 on the probe support 12 will be described later.

  The probe moving mechanism 100 moves the probe support 12. For example, the probe moving mechanism 100 moves the probe support 12 three-dimensionally in three axis directions (X axis, Y axis, Z axis) that are orthogonal to each other and include the vertical direction (Z axis).

  As shown in FIGS. 3 and 4, the probe moving mechanism 100 includes probe support mechanisms 110, 120, and 130 for movably supporting the probe support 12 in the three-axis directions, and the probe support 12 in the three directions. Motors 111, 121, 131 (drive devices) for moving in the axial direction are provided.

  The probe support mechanism 110 includes a rail 112 extending in the X-axis direction, a slider 113 that moves along the rail 112, a shaft 114 that is arranged in parallel to the rail 112, a bush 115 that moves along the shaft 114, and a slider. The arm 116 is arranged upright on the arm 113 and the arm 117 is arranged upright on the bush 115. The rail 112 and the shaft 114 are fixed with respect to the base surface 300.

  The motor 111, the rail 112, and the slider 113 constitute a belt-driven linear motion guide mechanism. The arms 116 and 117 are connected via a rail 122 and a shaft 124 of the probe support mechanism 120. By driving the motor 111, the slider 113, the bush 115, and the arms 116 and 117 are moved in the X-axis direction. The probe support mechanism 110 may be configured to move the arms 116 and 117 by a ball screw drive or a rack and pinion drive system.

  The probe support mechanism 120 includes a rail 122 extending in the Y-axis direction, a slider 123 that moves along the rail 122, a shaft 124 that is arranged in parallel to the rail 122, a bush 125 that moves along the shaft 124, and a slider 123 and a Z mount 126 connected to the bush 125. One end side of the rail 122 and the shaft 124 is supported by the arm 116 of the probe support mechanism 110, and the other end side is supported by the arm 117.

  The motor 121, the rail 122, and the slider 123 constitute a belt-driven linear motion guide mechanism. By driving the motor 121, the slider 123, the bush 125, and the Z mount 126 are moved in the Y-axis direction. The probe support mechanism 120 may be configured to move the Z mount 126 by a ball screw drive or a rack and pinion drive method.

  The probe support mechanism 130 includes a screw shaft 132 disposed along the Z-axis direction and a movable body 133 including a nut that moves along the screw shaft 132. The screw shaft 132 is rotatably supported on the Z mount 126. The probe support 12 is attached to the movable body 133.

  The Z mount 126 includes a motor 131, a pulley 134 connected to the rotation shaft of the motor 131, a pulley 135 connected to the upper end of the screw shaft 132, and a belt 136 that transmits the rotation of the pulley 134 to the pulley 135. Is installed.

  The movable body 133 and the probe support body 12 are moved in the Z-axis direction by driving the motor 131. For example, the height position of the center axis of the shaft 114 is set as the Z mount reference 127, and the height position of the Z mount 126 is adjusted based on the height position of the Z mount reference 127. The probe support mechanism 130 may be configured to move the probe support 12 by a belt driving method.

  A dispensing probe 8 is supported on the probe support 12. The dispensing probe 8 is supported on the probe support 12 so that the compression coil spring 141 is contracted so that the dispensing probe 8 can move up and down.

  A pre-punch pin 11 is supported on the probe support 12 via a pre-punch shaft 142, a shaft cover 143, and a locking mechanism 144 so as to be detachable. The upper end side of the pre-punch shaft 142 is slidably disposed in the pipe-shaped shaft cover 143.

  The shaft cover 143 is detachably locked to either the probe support 12 or the Z mount 126 by a locking mechanism 144. For example, the locking mechanism 144 is configured such that the main body is attached to the shaft cover 143, and the locking member movably attached to the main body is detachably connected to the probe support 12 or the Z mount 126, thereby The cover 143 is locked to either the probe support 12 or the Z mount 126.

  A sensor dog 145 used for detecting the presence / absence of a sample is fixed to the upper end side of the pre-punch shaft 142. The sensor dog 145 has a function of limiting the range of movement of the pre-punch shaft 142 downward by contacting the upper end surface of the shaft cover 143.

  The lower end side of the pre-punch shaft 142 is disposed in the compression coil spring 146. Due to the restoring force of the compression coil spring 146, the lower end surface of the compression coil spring 146 contacts the upper end surface of the pre-punch pin 11, the upper end surface of the compression coil spring 146 contacts the lower end surface of the shaft cover 143, and the pre-punch shaft 142 The sensor dog 145 fixed to the upper end side of the shaft is in contact with the upper end surface of the shaft cover 143. When the probe support 12 and the shaft cover 143 are simultaneously lowered by the locking mechanism 144 and the pre-punch pin 11 comes into contact with the upper end portion of the sample tube or the protective sheet, the compression coil spring 146 contracts, so that the pre-punch pin 11 becomes the shaft cover. 143 moves up and down and is recognized by the sample presence / absence detection sensor 150.

  Further, a stopper 147 for restricting the upward movement range of the pre-punch shaft 142 is fixedly attached to the pre-punch shaft 142 at a position between the pre-punch pin 11 and the shaft cover 143. The stopper 147 is disposed in the compression coil spring 146 so as not to interfere with movement by contacting the compression coil spring 146. In addition, you may provide the stopper which the upper end part of the pre punch shaft 142 contacts in the latching mechanism 144. FIG.

  The probe support 12 is provided with a sample presence / absence detection sensor 148. The sample presence / absence detection sensor 148 includes a slit in which a part of the sensor dog 145 is disposed, and detects whether or not a part of the sensor dog 145 is located in the slit, for example, optically or by a change in a magnetic field. The sample presence / absence detection sensor is not limited to the configuration using the sensor dog 45 and the sample presence / absence detection sensor 148, and may be, for example, a shock sensor capable of detecting a shock to the pre-punch pin 11.

  As shown in FIG. 2, the dispensing probe mechanism 6 also includes, for example, a metering pump unit 4, opening / closing valves 9 a and 9 b, and a cleaning liquid container 10. The dispensing probe 8 is connected to the metering pump 4 through an open / close valve 9a. The cleaning liquid container 10 is connected to the metering pump 4 via the open / close valve 9b. The cleaning liquid container 10 contains a cleaning liquid. The cleaning liquid is pure water, for example.

  Further, as shown in FIG. 1, the dispensing probe mechanism 6 includes a liquid level detection unit 3 for detecting the remaining amount of the separation buffer and the reagent. The liquid level detection unit 3 includes, for example, a sensor that detects the liquid level based on a change in capacitance of the entire dispensing probe 8.

  Next, the suction nozzle mechanism 19 will be described. The suction nozzle mechanism 19 sucks and removes the separation buffer, sample or cleaning liquid in the reservoir of the chip 5 and pressurizes the cleaning liquid or separation buffer into the flow path of the chip 5.

  The suction nozzle mechanism 19 includes a pressurized liquid feeding unit 16, suction nozzles 22-1 to 22-4 provided corresponding to the reservoirs of the chip 5, and a pressurized liquid feeding unit 16 and suction nozzles 22-1 to 22-4. And a nozzle guide mechanism 200 for guiding the moving direction of the nozzle support 17.

Referring also to FIG. 14, the pressurized liquid feeding unit 16 includes an air cylinder 161, and sucks and discharges air from a discharge port 162 provided on a surface facing the chip 5.
The suction nozzles 22-1 to 22-4 are connected to pumps provided in the suction pump unit 18, respectively.

  The nozzle guide mechanism 200 includes, for example, nozzle support mechanisms 210 and 220 that movably support the probe support 12 in two axial directions (Y-axis and Z-axis) including the vertical direction (Z-axis).

  The nozzle support mechanism 210 includes a rail 211 extending in the Y-axis direction, a slider 212 that moves along the rail 211, a shaft 213 that is arranged in parallel to the rail 211, a bush 214 that moves along the shaft 213, and a slider A table 215 for connecting 212 and the bush 214 is provided. The rail 211 and the shaft 213 are fixed with respect to the base surface 300.

  The nozzle support mechanism 220 includes two shafts 221 arranged upright on the table 215, and a bush 222 that moves along the shaft 221. The nozzle support 17 is attached to the bush 222. A stopper 223 is fixed to the upper end side of the shaft 221.

  A compression coil spring 224 is disposed along the outer periphery of the shaft 221 between the table 215 and the bush 222. Due to the restoring force of the compression coil spring 224, the lower end surface of the compression coil spring 224 contacts the upper surface of the table 215, the upper end surface of the compression coil spring 224 contacts the lower end surface of the bush 222, and the upper end surface of the bush 222 is the stopper 223. Touching. As the compression coil spring 224 contracts, the nozzle support 17 moves up and down with respect to the table 215.

A hole 401 into which the tip of the pre-punch pin 11 is fitted is provided on the upper surface of the nozzle support 17.
The suction nozzles 22-1 to 22-4 are held by the nozzle support 17 so that the tip position of the suction nozzles 22-1 to 22-4 can move toward the nozzle support 17 with respect to the nozzle support 17 by the compression coil spring 402 contracting. Yes. Although only the suction nozzle 22-1 is shown in FIG. 4, the tip positions of the suction nozzles 22-2 to 22-4 can be moved toward the nozzle support 17 as with the suction nozzle 22-1. It has become.

  2 and 3, the microchip electrophoresis apparatus 1 includes a probe cleaning unit 14 for cleaning the dispensing probe 8 and a nozzle for cleaning the suction nozzles 22-1 to 22-4. A cleaning unit 28 is provided. A cleaning liquid is supplied to the probe cleaning unit 14 and the nozzle cleaning unit 28.

  For example, a 96-well plate 13 containing a sample is disposed in the movement range of the dispensing probe 8. In addition, a container containing a reagent and a separation buffer is arranged in the reagent arrangement unit 15 provided in the movement range of the dispensing probe 8. Note that the reagent and the separation buffer may be accommodated in the holes of the well plate 13.

The voltage application unit 24 applies a predetermined voltage to each of the flow path ends of the chip 5.
The detection unit 31 detects, for example, fluorescence of the sample component separated by the separation channel in the chip 5. The detection unit 31 includes, for example, an LED (light emitting diode) 30 that irradiates a part of the separation channel with excitation light, and fluorescence generated by excitation of the sample component moving through the separation channel by the excitation light from the LED 30. Is provided.

  The detection unit 31 includes a photomultiplier tube 36 that receives the fluorescence through a filter 34 that removes the excitation light component from the fluorescence from the optical fiber 32 and transmits only the fluorescence component. The excitation light source is not limited to the LED, and for example, an LD (laser diode) may be used.

  The control unit 38 controls operations of the dispensing probe mechanism 6, the suction nozzle mechanism 19, the voltage application unit 24, and the detection unit 31. The control unit 38 is realized by, for example, a microcomputer equipped with a CPU (Central Processing Unit) and a storage device. The control unit 38 is connected to a computer 40 provided outside the microchip electrophoresis apparatus 1.

  The computer 40 is realized by, for example, a personal computer (PC) or a dedicated computer. The computer 40 is an external control device for instructing the operation of the microchip electrophoretic apparatus 1 and for capturing and processing data obtained by the detection unit 31.

  5 and 6 show an example of the chip 5. In addition, the chip (microchip) in the embodiment of the present invention refers to a chip having a channel formed inside and a reservoir opened at each channel end, and is not necessarily limited to a small size. Absent.

  As shown in FIG. 5, the chip 5 includes a pair of transparent substrates 51 and 52, for example. The transparent substrates 51 and 52 are made of, for example, quartz glass, other glass substrates, or resin substrates.

  On the surface of one substrate 52, as shown in (B), capillary grooves 54 and 55 for electrophoresis intersecting each other are formed. The other substrate 51 has through holes as reservoirs 53-1 to 53-4 at positions corresponding to the ends of the grooves 54 and 55, as shown in FIG. As shown in (C), the two substrates 51 and 52 are overlapped and joined to form the chip 5. By the capillary grooves 54 and 55, a separation channel 55 for electrophoresis separation of the sample and a sample introduction channel 54 for introducing the sample into the separation channel 55 are formed in the chip 5.

  The chip 5 is basically the one shown in FIG. 5, but in order to facilitate the handling, as shown in FIG. 6, an electrode terminal for applying a voltage is previously formed on the chip. use. FIG. 6 is a plan view of the chip 5.

  The four reservoirs 53-1 to 53-4 are also ports for applying a voltage to the flow paths 54 and 55. Ports # 1 and # 2 are ports located at both ends of the sample introduction channel 54. Ports # 3 and # 4 are ports located at both ends of the separation channel 55.

  In order to apply a voltage to each port, electrode patterns 56-1 to 56-4 are formed from the inner wall surface of the reservoirs 53-1 to 53-4 constituting each port to the surface of the chip 5. The electrode patterns 56-1 to 56-4 are connected to the voltage application unit 24 (see FIG. 2).

  In order to facilitate the handling of the chip 5, the peripheral portions of the three sides of the chip 5 are held by a resin-made chip frame 57 that is substantially U-shaped in plan view.

  The chip holding part and the cover will be described with reference to FIGS. FIG. 7 is a schematic plan view showing the chip holding portion in a state where the chip is mounted. FIG. 8 is a schematic plan view showing the cover.

  As shown in FIG. 7, the chip holding unit 7 includes a chip holder 71 for holding the chip 5 and a recess 74. Screw holes 72 for fixing the cover 26 are provided at two diagonal positions of the chip holding portion 7. As shown in FIG. 6, holes 62 through which the screws 27 pass are also provided at two diagonal positions of the cover 26.

  Moreover, the chip | tip holding | maintenance part 7 is provided with the escape hole 73 in which the suction nozzle 22-4 is inserted. The escape hole 73 is provided at a position shifted from the holding position of the chip 5 in the Y-axis direction (see FIG. 2).

  The cover 26 is made of, for example, resin, and includes through holes 64-1 to 64-4 at positions corresponding to the reservoirs 53-1 to 53-4 of the chip 5. Further, the cover 26 includes a clearance hole 63 formed of a through hole into which the suction nozzle 22-4 is inserted at a position corresponding to the clearance hole 73 of the chip holding portion 7.

  The inner diameters of the through holes 64-1 to 64-3 and the escape holes 63 are such that the suction nozzles 22-1 to 22-4 can be inserted. Moreover, the internal diameter of the through-hole 64-4 is a magnitude | size which can insert the pressurized liquid feeding part 16. FIG.

  Electrodes 66-1 to 66-4 that are electrically connected to the electrode patterns 56-1 to 56-4 of the chip 5 are provided on the back side of the cover 26. The electrodes 66-1 to 66-4 are connected to the terminal connection portion 67 by electric wiring. The terminal connection unit 67 is connected to the voltage application unit 24 (see FIG. 2).

  With reference to FIG. 4, the example of a movement of the pre punch pin 11, the dispensing probe 8, and the suction nozzle 22-1 is demonstrated.

  The pre-punch pin 11 is used for making a hole in the protective sheet 13a covering the sample storage portion of the well plate 13 (sample container), detecting the presence / absence of a sample, moving the nozzle support 17 and the like.

  In the punching operation for the protective sheet 13a, first, the motors 111 and 121 of the probe moving mechanism 100 are driven, and the Z-mount is performed so that the pre-punch pin 11 is disposed above the sample container to be punched in the well plate 13. 126 is moved. The motor 131 is driven in a state where the shaft cover 143 is locked to the probe support 12 by the locking mechanism 144, and together with the probe support 12, the pre-punch pin 11, the pre-punch shaft 142, the shaft cover 143, The stop mechanism 144, the sensor dog 145, and the sample presence / absence detection sensor 148 are lowered.

  When the tip of the pre-punch pin 11 comes into contact with the protective sheet 13 a and the probe support 12 is further lowered, the compression coil spring 146 contracts, and the probe support 12, the shaft cover 143, and the sample presence / absence detection sensor 148 are replaced with the well plate 13. Approached. At this time, the positions (height positions) of the pre-punch pin 11, the pre-punch shaft 142, and the sensor dog 145 with respect to the well plate 13 do not change. As a result, the sensor dog 145 is pushed up with respect to the probe support 12 and the sample presence / absence detection sensor 148 to leave the slit of the sample presence / absence detection sensor 148, and the presence of the protective sheet 13a (the presence of a sample) is detected.

  When the probe support 12 is further lowered, the compression coil spring 146 further contracts, the stopper 147 fixed to the pre-punch shaft 142 comes into contact with the shaft cover 143, and the pre-punch shaft 142 and the pre-punch pin 11 are lowered. A through hole is formed in the protective sheet 13a. After the through hole is formed, the probe support 12 is raised, the pre-punch pin 11 is moved above the sample container to be drilled next, and the protective sheet 13a is drilled and the presence / absence of the sample is detected.

  If the sample container or the protective sheet 13a is not disposed at the position where the pre-punch pin 11 is lowered, or if the protective sheet 13a has already been through-holed, the pre-punch pin 11 and the pre-punch shaft 142 and the sensor member 145 are not pushed up with respect to the probe support 12, the shaft cover 143, and the sample presence / absence detection sensor 148, and the sensor member 145 remains disposed in the slit of the sample presence / absence detection sensor 148. Thereby, it is detected that there is no sample at that position.

Next, the movement of the nozzle support 17 will be described.
In the movement operation of the nozzle support 17, first, the motors 111 and 121 are driven to move the Z mount 126 so that the pre-punch pin 11 is disposed above the hole 401 of the nozzle support 17. The motor 131 is driven in a state where the shaft cover 143 is locked to the probe support 12 by the locking mechanism 144, and the pre-punch pins 11 and the like are lowered together with the probe support 12.

  The tip of the pre-punch pin 11 is inserted into the hole 401. As a result, the nozzle support 17 is connected to the probe support 12 via the pre-punch pin 11, the pre-punch shaft 142, the shaft cover 143, and the locking mechanism 144.

  At this time, the probe support 12 may be lowered so that the stopper 147 does not contact the shaft cover 143, or the probe support 12 may be lowered so that the stopper 147 contacts the shaft cover 143. However, when the stopper 147 is brought into contact with the shaft cover 143, the restoring force of the compression coil spring 146 and the restoring force of the compression coil spring 224 are such that the probe support 12 does not descend due to the restoring force of the compression coil spring 146. Selected.

  With the tip of the pre-punch pin 11 inserted into the hole 401 and the nozzle support 17 connected to the probe support 12, the motor 111 of the probe moving mechanism 100 is driven to move the Z mount 126 in the Y-axis direction. Is done. Accordingly, the probe support 12 and the nozzle support 17 are moved in the Y-axis direction, and the nozzle support 17 is disposed at a desired position. For example, the Z mount 126 and the nozzle support so that the suction nozzles 22-1 to 22-3 and the pressurized liquid feeding unit 16 are disposed on the chip 5 and the suction nozzle 22-4 is disposed on the escape hole 73. 17 is moved.

  With the nozzle support 17 connected to the probe support 12, the motor 131 of the probe moving mechanism 100 is driven, the pre-punch pins 11 and the like are lowered together with the probe support 12, the nozzle support 17 is pushed down, and the nozzle The suction nozzles 22-1 to 22-4 and the pressurized liquid feeding unit 16 are lowered together with the support 17 by a predetermined amount. At this time, in order to lower the nozzle support 17 by an accurate amount, the restoring force of the compression coil spring 146 and the restoring force of the compression coil spring 224 are maintained so that the stopper 147 is kept in contact with the shaft cover 143. It is preferable to be selected.

  After a desired process is performed in a state where the nozzle support 17 is lowered, the motor 131 is driven to raise the probe support 12 by a predetermined amount, whereby the nozzle support is supported by the restoring force of the compression coil spring 224. 17 goes up. Thereafter, the motor 121 is driven to move the probe support 12 and the nozzle support 17 in the Y-axis direction.

  Thus, the suction nozzles 22-1 to 22-4 and the pressurized liquid feeding unit 16 are moved by driving the probe moving mechanism 100 in a state where the nozzle support 17 is detachably connected to the probe support 12. The microchip electrophoresis apparatus 1 does not include a dedicated drive device for moving the nozzle support 17. This eliminates the need for a drive shaft for moving the suction nozzles 22-1 to 22-4 in, for example, the horizontal direction (Y-axis direction) and the vertical direction (Z-axis direction), and thus the configuration of the microchip electrophoresis apparatus 1 is eliminated. Simplification and miniaturization can be achieved.

  Further, since the positioning accuracy of the Z mount 126 (positioning of the dispensing probe 8 in the XYZ-axis directions) becomes the positioning accuracy of the suction nozzles 22-1 to 22-4 as they are, the suction nozzles 22-1 to 22-4 are moved. As compared with the case where the drive shaft for this is provided, positioning adjustment becomes easy.

Next, the movement of the dispensing probe 8 will be described.
The dispensing probe 8 is used for sucking and dispensing a sample, a reagent, a separation buffer, dispensing a cleaning liquid, and the like. When the dispensing probe 8 is used, first, the motors 111 and 121 of the probe moving mechanism 100 are driven to place the dispensing probe 8 at a desired position, for example, above the target sample container of the well plate 13. Thus, the Z mount 126 is moved.

  The motor 131 is driven in a state where the shaft cover 143 is locked to the Z mount 126 by the locking mechanism 144, and the dispensing probe 8 is lowered together with the probe support 12. At this time, the pre-punch pin 11, the pre-punch shaft 142, the shaft cover 143, etc. are not lowered. Thus, the dispensing probe 8 is lowered to a desired position without being obstructed by the pre-punch pin 11 or the like. After the dispensing probe 8 is brought into contact with the bottom of the sample container of the well plate 13, when the probe support 12 is further lowered, the compression coil spring 141 is contracted, and the sensor dog 150 detects the presence / absence of the tip / sample container bottom detection. The bottom position of the sample container is detected away from the slit of the sensor 150. As a result, it is possible to reliably suck a small amount of sample in the sample container.

  The operation of the microchip electrophoresis apparatus 1 will be described with reference to FIGS. In the microchip electrophoresis apparatus 1, the chip 5 is repeatedly used while being fixed to the chip holder 7 without being moved. The operation described here is a series of steps of washing the chip used in the previous analysis, filling the flow path and each reservoir with a separation buffer, then dispensing the sample, performing electrophoresis, and performing analysis. is there. These operations are performed under the control of the control unit 38 of the microchip electrophoresis apparatus 1. 9 to 13, the cover 26 is not shown. 9 to 13, the illustration of the pressurized liquid feeding unit 16 is omitted except where it is necessary to explain. Moreover, (A)-(U) of FIGS. 9-13 respond | corresponds to process (A)-(U) demonstrated below.

  (A) shows one chip 5. The chip 5 is equivalent to that shown in FIGS. The separation channel 55 and the sample introduction channel 54 are provided so as to intersect with each other, and reservoirs 53-1 to 53-4 are formed at the ends of the channels 54 and 55, respectively. Here, the chip 5 is in a state where the analysis of the previous sample is completed, and the separation buffer and the sample solution remain in the flow path and each reservoir. Further, the separated sample remains in the separation buffer in the flow channel 55.

  (B) First, in order to remove the sample solution stored in the sample storage reservoir 53-1, the nozzle support 17 is moved down onto the chip 5, and then the suction nozzle 22-1 is moved. It is inserted into the reservoir 53-1 through the through hole 64-1 (see FIG. 8). As described above, the suction nozzles 22-1 to 22-4 are moved by driving the motors 121 and 131 of the probe moving mechanism 100 while the nozzle support 17 is detachably connected to the probe support 12. Is called.

  Further, as shown in FIG. 2, the length of the suction nozzle 22-1 is longer than that of the suction nozzles 22-2 and 22-3. The length of the suction nozzle 22-4 is longer than that of the suction nozzle 22-1. Moreover, the lower end surface of the pressurized liquid feeding part 16 is arrange | positioned rather than the front-end | tip of the suction nozzles 22-2 and 22-3 at the nozzle support body 12 side. Further, the suction nozzles 22-1 to 22-4 and the pressurized liquid feeding unit 16 are connected to the reservoirs 53-1 to 53-4 (through holes 64-1 to 64-4) is held by the nozzle support 17 so that the suction nozzle 22-4 is arranged above the escape holes 63, 73 (see also FIGS. 7 and 8). ing.

  With the configuration of the suction nozzles 22-1 to 22-4 and the pressurized liquid feeding unit 16, the tip of the suction nozzle 22-1 is inserted into the reservoir 53-1, and pressed against the bottom of the reservoir 53-1, so that the suction nozzle It is possible to form a state in which the tips of 22-2 and 22-3 are not inserted into the reservoirs 53-2 and 53-3, and the tip of the suction nozzle 22-4 is inserted into the escape holes 63 and 73. . Further, the lower end surface of the pressurized liquid feeding part 16 is not in contact with the chip 5.

  In this state, when the suction pump unit 18 (see FIG. 2) is operated, the separation buffer in the reservoir 53-1 is removed by suction through the suction nozzle 22-1. At this time, in the suction pump unit 18, for example, only the pump connected to the suction nozzle 22-1 is operated.

  In this state, the suction nozzles 22-2 and 22-3 and the tip of the pressurized liquid feeding unit 16 are located inside the through holes 64-2, 64-3 and 64-4 (see FIG. 8). It may be arranged above the through holes 64-2, 64-3, 64-4.

  Further, the tip of the suction nozzle 22-4 inserted into the escape hole 73 may or may not be in contact with the bottom of the escape hole 73. However, considering the contamination of the tip of the suction nozzle 22-4, it is preferable that the tip of the suction nozzle 22-4 does not contact the bottom of the escape hole 73. The escape hole 73 may have a bottom or a through hole.

  (C) After the suction nozzles 22-1 to 22-4 are moved to the nozzle cleaning unit 28 and at least the suction nozzle 22-1 is cleaned, the dispensing probe 8 passes through the through-hole 64-1 (see FIG. 8). Is inserted into the reservoir 53-1. Then, a predetermined amount of cleaning liquid is supplied from the dispensing probe 8 to the reservoir 53-1. The supply of the cleaning liquid from the dispensing probe 8 will be described with reference to FIG. 2. When the opening / closing valve 9 a is closed and the opening / closing valve 9 b is opened, the metering pump 4 is aspirated to bring the cleaning liquid container 10 into the metering pump 4. Then, after the cleaning liquid is sucked, the open / close valve 9b is closed, the open / close valve 9a is switched to the open state, and the metering pump 4 is discharged.

(D) After the dispensing probe 8 is moved, the suction nozzle 22-1 is again inserted into the reservoir 53-1, and the cleaning liquid is sucked and removed.
(E) The cleaning liquid is supplied again from the dispensing probe 8 to the reservoir 53-1.

  (F) The suction nozzles 22-1 to 22-3 are inserted into the reservoirs 53-1 to 53-3 through the through holes 64-1 to 64-3 (see FIG. 8). After the nozzle support 17 is lowered and the tip of the suction nozzle 22-1 is pressed against the bottom of the reservoir 53-1, the nozzle support 17 is further lowered and the suction nozzles 22-2 and 22-3 are moved. The tip is pressed against the bottom of the reservoirs 53-2 and 53-3. The suction nozzle 22-4 is inserted into the escape holes 63 and 73. At this time, the lower end surface of the pressurized liquid feeding unit 16 may be disposed with a space from the chip 5 or may be in contact with the chip 5.

  The suction pump unit 18 is operated, and the cleaning liquid in the reservoir 53-1 and the separation buffer in the reservoirs 53-2 and 53-3 are removed by suction through the suction nozzles 22-1 to 22-3. At this time, in the suction pump unit 18, for example, only pumps connected to the suction nozzles 22-1 to 22-3 are operated.

  (G) The nozzle support 17 is moved, the suction nozzle 22-4 is inserted into the reservoir 53-4 via the through hole 64-4 (see FIG. 8), and the tip of the suction nozzle 22-4 is the reservoir Pressed against the bottom of 53-4. At this time, the suction nozzles 22-1 to 22-3 are not inserted into the reservoirs 53-1 to 53-3. The suction nozzles 22-1 to 22-3 and the tip of the pressurized liquid feeding section 16 may be disposed above the cover 26, or a relief hole is separately provided in the cover 26 so as to be inserted into the relief hole. Also good.

  In a state where the suction nozzle 22-4 is inserted into the reservoir 53-4, the suction pump unit 18 is operated, and the separation buffer in the reservoir 53-4 is removed by suction through the suction nozzle 22-4. At this time, in the suction pump unit 18, for example, only the pump connected to the suction nozzle 22-4 is operated.

(H) A predetermined amount of cleaning liquid is supplied from the dispensing probe 8 to the reservoir 53-4.
(I) The nozzle support 17 is moved, the suction nozzles 22-1 to 22-3 are inserted into the reservoirs 53-1 to 53-3, and the lower end surface of the pressurized liquid feeding unit 16 is located around the reservoir 53-4. It contacts the surface of the chip 5. Air is pressurized and supplied from the pressurized liquid feeding unit 16 to the reservoir 53-4, and the cleaning liquid in the reservoir 53-4 is pressurized and fed into the flow paths 54 and 55 to clean the flow paths 54 and 55. The liquid in the flow paths 54 and 55 is pushed out to the reservoirs 53-1 to 53-3. In addition, the cleaning liquid supplied into the flow paths 54 and 55 is supplied to the reservoir 53-4 from the pressurized liquid feeding section 16 with an excessive amount of air with respect to the capacity of the reservoir 53-4. , 55 is pushed out to the reservoirs 53-1 to 53-3. The liquid pushed out to the reservoirs 53-1 to 53-3 is removed by suction through the suction nozzles 22-1 to 22-3.

FIG. 14 is a side view schematically showing a partial cross-section of the connection state between the pressurized liquid feeding section 16 and the chip 5.
The pressurized liquid feeding unit 16 includes a syringe pump 161 and sucks and discharges air from a discharge port 162 provided on the lower end surface. An airtight holding member made of an elastic body, for example, an O-ring 163, is provided on the lower end surface of the pressurized liquid feeding unit 16 so as to be in close contact with the surface of the chip 5 around the reservoir 53-4. Further, the pressurized liquid feeding section 16 is held by the nozzle support 17 so that the lower end surface position can be moved toward the nozzle support 17 with respect to the nozzle support 17 by the compression coil spring 403 contracting.

  By pressing the lower end surface of the pressurized liquid feeding unit 16 around the reservoir 53-4 of the chip 5, the syringe pump 161 of the pressurized liquid feeding unit 16 is attached to the flow path 55 of the chip 5 while keeping airtightness. The O-ring 163 is pressed against the chip 5 while the lower end surface of the pressurized liquid feeding unit 16 is spaced from the chip 5, so that airtightness between the pressurized liquid feeding unit 16 and the chip 5 can be secured. It is.

  The operations of the steps (H) and (I) or the operations of the steps (G) to (I) are repeated a plurality of times, for example, 12 times, so that the inside of the channels 54 and 55 of the chip 5 and the reservoirs 53-1 to 53- The inside of 4 is cleaned.

  (J) After the channels 54 and 55 and the reservoirs 53-1 to 53-4 are cleaned, the suction nozzle 22-4 is inserted into the reservoir 53-4, and the cleaning liquid remaining in the reservoir 53-4 Is removed by suction.

  (K) After the separation buffer is sucked into the dispensing probe 8, the dispensing probe 8 is inserted into the reservoir 53-4. A predetermined amount of separation buffer is discharged from the dispensing probe 8 and dispensed into the reservoir 53-4.

  (L) In the same manner as in the above step (I), the suction nozzles 22-1 to 22-3 are inserted into the reservoirs 53-1 to 53-3, and the lower end surface of the pressurized liquid feeding unit 16 is positioned around the reservoir 53-4. It contacts the surface of the chip 5. Air is pressurized and supplied from the pressurized liquid feeding unit 16 to the reservoir 53-4, and the separation buffer in the reservoir 53-4 is pressurized and fed into the flow paths 54 and 55 and filled. An excessive amount of the separation buffer flows into the reservoirs 53-1 to 53-3 and is removed by suction through the suction nozzles 22-1 to 22-3. The amount of air supplied under pressure from the pressurized liquid feeding section 16 to the reservoir 53-4 is adjusted so that the separation buffers are filled in the flow paths 54 and 55.

  (M) After the separation buffer is filled in the flow paths 54 and 55, the suction nozzle 22-4 is inserted into the reservoir 53-4, and the separation buffer remaining in the reservoir 53-4 is removed by suction. As a result, the separation buffer remains in the flow paths 54 and 55 only.

  (N) to (Q) The dispensing probe 8 sequentially dispenses a predetermined amount of separation buffer into the reservoirs 53-1 to 53-4. The order in which the separation buffers are dispensed into the reservoirs 53-1 to 53-4 is not limited to the order of the steps (N) to (Q), and is not particularly limited.

  (R) The reservoirs 53-1 to 53-4 are provided by the voltage application unit 24 via the electrode patterns 56-1 to 56-4 and the electrodes 66-1 to 66-4 (see FIGS. 2, 6, and 8). A predetermined test voltage is applied to the electrophoretic test. In this migration test, for example, it is confirmed whether dust or bubbles are mixed in the flow path by detecting a current value between the electrode patterns. Here, the test voltage may be the same as the migration voltage for separating the sample, but may be a lower voltage than that.

  If it is determined in this electrophoresis test step that the separation buffer is normally filled into the flow path, the process proceeds to the next step (S) in order to inject the sample for analysis. If it is not determined that the separation buffer is normally filled into the flow path, the process returns to step (J) to refill the flow path with the separation buffer.

  For example, the number of times that the refilling of the separation buffer into the flow paths 54 and 55 is allowed is set in advance. If it is not determined that the separation buffer has been normally filled into the flow paths 54 and 55 even if the separation buffer is refilled the same number of times, the chip 5 is removed from the chip holding unit 7 and another chip 5 is removed. Replace with. The number of times that the refilling of the separation buffer is allowed is not particularly limited, but for example, 2 or 3 times is appropriate.

  (S) The suction nozzle 22-1 is inserted into the sample storage reservoir 53-1, and the separation buffer in the reservoir 53-1 is removed by suction.

  (T) A predetermined amount of sample solution is dispensed from the dispensing probe 8 into the reservoir 53-1. The dispensing amount of the sample solution is an amount set for each chip 5.

  (U) The reservoirs 53-1 to 53-4 are connected by the voltage application unit 24 via the electrode patterns 56-1 to 56-4 and the electrodes 66-1 to 66-4 (see FIGS. 2, 6, and 8). A predetermined voltage for introducing the sample is applied to the sample, and the sample is guided to the intersection of the flow paths 54 and 55 (see FIG. 6). Thereafter, the applied voltage is switched to a predetermined voltage for electrophoretic separation, and the sample is guided toward the reservoir 53-4 into the separation channel 55 and separated in the separation channel 55. The separated sample is detected by the detection unit 31 (see FIG. 2).

  Although one embodiment of the present invention has been described above, the configuration, arrangement, numerical values, materials, and the like in the embodiment are examples, and the present invention is not limited to this, and the present invention described in the claims. Various modifications are possible within the scope of the invention.

  For example, the method of detachably connecting the nozzle support 17 and the probe support 12 is not limited to the method using the pre-punch pin 11, and any method may be used. For example, the nozzle support 17 is provided with a protrusion or a recess, and the probe support 12 is provided with a recess or a protrusion fitted to the nozzle support 17, and the nozzle support 17 and the probe support 12 are formed using the protrusion and the recess. You may connect so that attachment or detachment is possible.

  In the above embodiment, one chip 5 is arranged in the microchip electrophoresis apparatus 1, but the number of chips arranged in one microchip electrophoresis apparatus may be two or more. When a plurality of chips are arranged in one microchip electrophoresis apparatus, the chips are arranged along the moving direction of the nozzle support guided by the nozzle guide mechanism.

  Further, the chip used in the microchip electrophoresis apparatus of the embodiment of the present invention is not limited to the configuration shown in FIGS. 5 and 6, and has a separation channel for separating a sample by electrophoresis inside. Any chip may be used as long as it has at least a flow path and has a reservoir opened at each end of the flow path. For example, the chip may have a configuration in which the sample introduction channel 54 and the reservoirs 54-1 and 54-2 are not provided for the chip 5.

1 Microchip electrophoresis device 5 Chip (microchip)
7 Tip holder 8 Dispensing probe 12 Probe support 11 Pre-punch pin 17 Nozzle supports 22-1 to 22-4 Suction nozzles 53-1 to 53-4 Reservoir 54 Channel 55 Separation channel 100 Probe moving mechanism 110 , 120, 130 Probe support mechanism 111, 121, 131 Drive device 200 Nozzle guide mechanism 210, 220 Nozzle support mechanism

Claims (4)

A chip holding part for holding a microchip having a flow path including at least a separation flow path for separating a sample by electrophoresis inside and having a reservoir opened at each end of the flow path;
A probe support for supporting a dispensing probe for dispensing liquid into the reservoir;
A nozzle support for supporting a suction nozzle for sucking the liquid in the reservoir;
A moving mechanism for moving any one of the probe support and the nozzle support; and
A microchip electrophoresis apparatus in which the probe support and the nozzle support are detachably connected to each other, and the supports are moved simultaneously by the moving mechanism.   2. The micro of claim 1, wherein one of the probe support and the nozzle support includes a pin provided so as to extend vertically downward, and the two supports are connected via the pin. Chip electrophoresis device. The pin is a pre-punch pin provided so as to extend downward from the probe support and is used to make a hole in the protective sheet of the sample container. The hole into which the tip of the pre-punch pin is inserted into the nozzle support The microchip electrophoresis apparatus according to claim 2, wherein the support is connected by inserting the pre-punch pin into the hole. A nozzle guide mechanism for guiding the moving direction of the nozzle support;
The probe moving mechanism includes a probe support mechanism that movably supports the probe support in three axial directions including a vertical direction perpendicular to each other, and a drive device for moving the probe support in the three axial directions. Prepared,
The microchip electrophoresis apparatus according to claim 3, wherein the nozzle guide mechanism includes a nozzle support mechanism that movably supports the nozzle support in two axial directions including the vertical direction out of the three axial directions.

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