WO2024070314A1 - Dispositif d'électrophorèse, procédé de commande pour dispositif d'électrophorèse et programme de commande pour dispositif d'électrophorèse - Google Patents

Dispositif d'électrophorèse, procédé de commande pour dispositif d'électrophorèse et programme de commande pour dispositif d'électrophorèse Download PDF

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Publication number
WO2024070314A1
WO2024070314A1 PCT/JP2023/029825 JP2023029825W WO2024070314A1 WO 2024070314 A1 WO2024070314 A1 WO 2024070314A1 JP 2023029825 W JP2023029825 W JP 2023029825W WO 2024070314 A1 WO2024070314 A1 WO 2024070314A1
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Prior art keywords
flow path
channel
concentrated layer
voltage
path end
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PCT/JP2023/029825
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English (en)
Japanese (ja)
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直人 磯▲崎▼
大亮 衛藤
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富士フイルム株式会社
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Publication of WO2024070314A1 publication Critical patent/WO2024070314A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis

Definitions

  • This disclosure relates to an electrophoretic device, a control method for an electrophoretic device, and a control program for an electrophoretic device.
  • Electrophoresis devices using microchannel chips are known. For example, a technique is known in which isotachophoresis is performed as electrophoresis to concentrate a sample and form a concentrated layer, and then the isotachophoresis is switched to capillary electrophoresis to separate specific components contained in the concentrated layer (see, for example, JP 2019-158520 A).
  • the reagent state at the start of capillary electrophoresis was not always constant. For example, if the timing for switching to capillary electrophoresis is set to a certain time after the concentrated layer reaches a certain position, if the moving speed of the concentrated layer changes for each measurement, the actual position of the concentrated layer at the time of switching will also change for each measurement. As a result, the reagent state at the start of capillary electrophoresis is not constant.
  • the present disclosure provides an electrophoresis device that can keep the reagent state constant at the start of capillary electrophoresis, a control method for the electrophoresis device, and a control program for the electrophoresis device.
  • the first aspect of the present disclosure is an electrophoresis device using a microchannel chip including at least a first branch channel connected to a first channel end, a second branch channel connected to a second channel end, and a third branch channel connected to a third channel end, which are branched at a branch point, and in which a channel through which a sample moves by electrophoresis is formed by applying a voltage to a pair of channel ends selected from the first channel end, the second channel end, and the third channel end, and the electrophoresis device is provided with at least one processor, which applies a first voltage for isotachophoresis to the first channel end and the third channel end, moves the sample in the first branch channel toward the third channel end by isotachophoresis, thereby forming a concentrated layer of the sample, and when the concentrated layer reaches a first position set in advance in the third branch channel, stops applying the first voltage to the first channel end and the third channel end, and applies a second voltage for capillary electrophoresis to the second
  • the processor may obtain a detection signal indicating that the concentrated layer has reached the second position from a detection device that detects that the concentrated layer has reached a second position set closer to the first flow path end than the first position in a path along which a sample moves by isotachophoresis from a first flow path end via a branch point, derive a movement speed of the concentrated layer passing through the second position based on the detection signal, derive a movement time required for the concentrated layer to move from the second position to the first position based on the movement speed, and, when the movement time has elapsed since the concentrated layer reached the second position, perform control to stop application of a first voltage to the first flow path end and the third flow path end and apply a second voltage to the second flow path end and the third flow path end.
  • the processor may obtain a detection result from a detection device that detects the position of the concentrated layer, and when the detection result indicates that the concentrated layer has reached a first position, control is performed to stop application of a first voltage to the first flow path end and the third flow path end, and to apply a second voltage to the second flow path end and the third flow path end.
  • the detection device may detect the position of the concentrated layer optically or magnetically, or may detect it based on a change in electric potential.
  • the fifth aspect of the present disclosure is a method for controlling an electrophoresis device, the method using a micro-channel chip in which a channel is formed in which a sample moves by electrophoresis by applying a voltage to a pair of channel ends selected from the first channel end, the second channel end, and the third channel end, the channel including at least a first branch channel connected to a first channel end, a second branch channel connected to a second channel end, and a third branch channel connected to a third channel end, the channel branching at a branch point, the voltage being applied to a pair of channel ends selected from the first channel end, the second channel end, and the third channel end, the electrophoresis device is provided with a processor, the processor applies a first voltage for isotachophoresis to the first channel end and the third channel end, and moves the sample in the first branch channel toward the third channel end by isotachophoresis to form a concentrated layer of the sample, and when the concentrated layer reaches a first position in the third branch channel that is set
  • the sixth aspect of the present disclosure is a control program for an electrophoresis device, the control program being for causing a computer to function to control an electrophoresis device using a microchannel chip in which a channel is formed in which a sample moves by electrophoresis by applying a voltage to a pair of channel ends selected from the first channel end, the second channel end, and the third channel end, the channel including at least a first branch channel connected to a first channel end, a second branch channel connected to a second channel end, and a third branch channel connected to a third channel end, the control program controlling the electrophoresis device to apply a first voltage for isotachophoresis to the first channel end and the third channel end, move the sample in the first branch channel toward the third channel end by isotachophoresis, thereby forming a concentrated layer of the sample, and when the concentrated layer reaches a first position in the third branch channel that is set in advance, stop applying the first voltage to the first channel end and the third channel end, and apply a
  • the electrophoresis device, the control method for the electrophoresis device, and the control program for the electrophoresis device disclosed herein can keep the reagent state constant at the start of capillary electrophoresis.
  • FIG. 2 is a schematic diagram showing an example of a configuration of a microchannel chip according to an exemplary embodiment.
  • FIG. 1 is a configuration diagram showing an example of the configuration of an electrophoretic device according to a first exemplary embodiment.
  • FIG. 1 is a schematic diagram showing an example of a microchannel chip according to an exemplary embodiment.
  • FIG. 2 is a block diagram illustrating an example of a hardware configuration of a control device according to an exemplary embodiment.
  • FIG. 2 is a functional block diagram illustrating an example of a configuration of a control device according to an exemplary embodiment.
  • FIG. 13 is a diagram for explaining a method for detecting the timing when the concentrated layer reaches a branch point.
  • FIG. 5 is a flowchart showing an example of a control process executed by a processor of the first exemplary embodiment.
  • FIG. 13 is a diagram illustrating an example of the configuration of an electrophoretic device according to a first modified example.
  • FIG. 11 is a functional block diagram showing an example of a configuration of a control device according to a first modified example. 13 is a flowchart showing an example of a control process executed by a processor in the first modification;
  • FIG. 13 is a configuration diagram showing an example of the configuration of an electrophoretic device according to a second exemplary embodiment.
  • 10 is a flowchart showing an example of a control process executed by a processor of the second exemplary embodiment.
  • FIG. 1 shows a schematic diagram of the flow channels in the microchannel chip 20 of this exemplary embodiment.
  • the micro-channel chip 20 of this exemplary embodiment includes channel ends 22 1 - 22 6 and channel ends 23 1 - 23 4.
  • Branch channels are connected to each of the channel ends 22 1 - 22 6 and the channel ends 23 1 - 23 4 , and are connected to a main channel 28 0 via the branch channels.
  • the channel ends 22 1 - 22 6 are reagent introduction sections for introducing reagents.
  • the reference numerals 1 - 6 used to distinguish between them will be omitted and they will be referred to collectively as "channel end 22.”
  • the flow channel ends 22 1 , 22 2 , and 22 3 are used to introduce a buffer solution for electrophoresis.
  • the flow channel end 22 4 is used to introduce a first labeled antibody solution (e.g., a solution containing a DNA-labeled tumor marker antibody).
  • the flow channel ends 22 5 and 22 6 are used to introduce an immune reaction solution between a tumor marker sample and a second labeled antibody solution (e.g., a solution containing a fluorescently labeled tumor marker antibody).
  • An electrode (not shown) is disposed at some of the flow path ends 22 1 to 22 6 (in this exemplary embodiment, flow path ends 22 1 to 22 3 ).
  • cathodes are disposed at the flow path ends 22 1 and 22 2
  • an anode is disposed at the flow path end 22 3.
  • a migration flow path is formed by the main flow path 28 0 , the branch flow path 28 1 connected to the flow path end 22 1 , the branch flow path 28 13 connected to the flow path end 22 2 , and the branch flow path 28 9 connected to the flow path end 22 3 .
  • Channel end 221 is connected to channel end 223 via main channel 280, branch channel 281 , and branch channel 289.
  • Channel end 222 is connected to channel end 223 via branch channel 2813 and branch channel 289.
  • Channel ends 224 to 226 are connected to main channel 280 via their respective branch channels.
  • the channel end 221 side of main channel 280 is referred to as the "upstream side, " and the channel end 223 side is referred to as the "downstream side.”
  • channel end 224 , channel end 225 , channel end 226 , and channel end 222 are arranged in this order from the upstream side to the downstream side.
  • the flow path ends 23 1 to 23 4 are waste liquid storage sections for storing waste liquid.
  • the reference numerals 1 to 4 for distinguishing between them will be omitted and they will be referred to collectively as "flow path end 23.”
  • the flow path end 23-1 is a portion (waste reservoir) for storing an excess of the buffer solution introduced from the flow path end 22-1 and an excess of the first labeled antibody solution introduced from the flow path end 22-4 .
  • the flow path end 23-2 is a portion (waste reservoir) for storing an excess of the first labeled antibody solution introduced from the flow path end 22-4 and an excess of the immune reaction solution introduced from the flow path end 22-5 .
  • the flow path end 23-3 is a portion (waste reservoir) for storing an excess of the immune reaction solution introduced from the flow path ends 22-5 and 22-6 .
  • the flow path end 23-4 is a portion (waste reservoir) for storing an excess of the immune reaction solution introduced from the flow path end 22-6 and an excess of the buffer solution introduced from the flow path ends 22-2 and 22-3 .
  • the flow path ends 23 1 to 23 4 are connected to the main flow path 28 0 via branch flow paths. These are connected in the order of flow path ends 23 1 , 23 2 , 23 3 , and 23 4 from the upstream side to the downstream side. More specifically, the flow path end 23 1 is disposed downstream of the flow path end 22 1 and upstream of the flow path end 22 4 .
  • the flow path end 23 2 is disposed downstream of the flow path end 22 4 and upstream of the flow path end 22 5 .
  • the flow path end 23 3 is disposed downstream of the flow path end 22 5 and upstream of the flow path end 22 6 .
  • the flow path end 23 4 is connected downstream of the flow path end 22 6 and upstream of the flow path end 22 2 .
  • the migration channel of this exemplary embodiment has, from the upstream side to the downstream side, a sample concentration region 30 and a sample separation region 32.
  • a buffer solution for electrophoresis is introduced upstream of the sample concentration region 30.
  • the sample concentration region 30 is a region in which a tumor marker, which is a sample, is concentrated using immune reactions and isotachophoresis (ITP), and in the example shown in FIG. 1, is provided from the branch point of the main channel 28-0 and the channel end 23-2 to the branch point of the main channel 28-0 and the channel end 22-2 .
  • ITP isotachophoresis
  • a voltage is applied between the channel end 22-1 and the channel end 22-3 .
  • the sample separation region 32 is a region in which a tumor marker, which is a sample, is separated from other components using capillary electrophoresis (CE), more specifically, capillary zone electrophoresis (CZE), and in the example shown in FIG. 1, is provided from the channel end 22-2 to the channel end 22-3 .
  • CE capillary electrophoresis
  • CZE capillary zone electrophoresis
  • the sample separation region 32 also includes a detection region 34 located downstream for detecting the sample that has migrated through the migration flow path.
  • the tumor marker sample can be analyzed as follows. After each reagent is introduced, when a voltage is applied between the channel end 221 and the channel end 223 , the first labeled antibody (DNA-labeled tumor marker antibody) introduced from the channel end 224 moves in the main channel 280 toward the channel end 223 according to the principle of isotachophoresis. In the sample concentration region 30, the first labeled antibody is concentrated, and an immune complex is formed between the tumor marker, the first labeled antibody, and the second labeled antibody (fluorescently labeled tumor marker antibody), forming a concentrated layer of the sample. The concentrated layer of the immune complex moves toward the channel end 223 and reaches the sample separation region 32. The unreacted (free) second labeled antibody does not have an electric charge in its molecule, so it remains at that position and does not reach the sample separation region 32.
  • the first labeled antibody DNA-labeled tumor marker antibody
  • the second labeled antibody fluorescently labeled tumor marker antibody
  • the electrodes are switched and a voltage is applied between the flow path end 222 and the flow path end 223.
  • the immune complex and the unreacted (free) first labeled antibody move toward the flow path end 223 at their respective moving speeds according to their charge and molecular size in the sample separation area 32.
  • the concentration of the tumor marker can be measured from the peak area of the fluorescence intensity when the immune complex containing the tumor marker as the core and the second labeled antibody having a fluorescent dye reaches the detection area 34. Note that the unreacted first labeled antibody does not have a fluorescent dye in its molecule, so it does not affect the fluorescence intensity even when it reaches the detection area 34, and does not affect the measurement of the concentration of the tumor marker.
  • the electrophoresis device 1 includes a control device 10, a power supply device 12, a detection device 14, and a micro-channel chip 20.
  • the micro-channel chip 20 shown in FIG. 2 and FIG. 3 includes three channel ends 25 (first channel end 25 1 to third channel end 25 3 ) used when applying a voltage in isotachophoresis and capillary electrophoresis.
  • a first branch channel 24 1 connected to the first channel end 25 1 , a second branch channel 24 2 connected to the second channel end 25 2 , and a third branch channel 24 3 connected to the third channel end 25 3 are connected by a branch point 26.
  • the first branch channel 24 1 is provided with the above-mentioned sample concentration region 30 and is used as a channel for sample concentration.
  • the third branch channel 24 3 is provided with the above-mentioned sample separation region 32 and detection region 34 and is used as a channel for sample separation and sample detection.
  • the branch flow path 24 3 (sample separation region 32 ) is provided with a switching position 28 for determining the timing to switch from isotachophoresis to capillary electrophoresis.
  • the power supply device 12 is a power source for applying a voltage to each of the first flow path end 25-1 to the third flow path end 25-3 under the control of the control device 10. Specifically, the power supply device 12 applies a voltage to electrodes (not shown) provided at each of the first flow path end 25-1 to the third flow path end 25-3 , but in this exemplary embodiment, it is simply said that the power supply device 12 "applies a voltage to the flow path end 25", etc.
  • the detection device 14 includes a sensor 15.
  • the sensor 15 is used to measure the concentration of a tumor marker and is a sensor for optically detecting an immune complex.
  • the sensor 15 is disposed at a position facing a detection point 29 in the detection region 34 of the third branch flow channel 24 3.
  • the sensor 15 includes, for example, a laser diode (LD) or a light-emitting diode (LED) that irradiates excitation light.
  • LD laser diode
  • LED light-emitting diode
  • the fluorescence generated by excitation with the excitation light irradiated from the sensor 15 (15 1 to 15 3 , see FIG. 8 and FIG. 11 ) is received by a photodetector such as a photodiode (PD) or a photomultiplier tube (PMT).
  • the detection device 14 outputs a detection signal corresponding to the fluorescence received as a detection result of the sensor 15 to the control device 10.
  • the control device 10 has the function of performing general control related to electrophoresis. Specifically, the control device 10 controls the electrodes that apply voltage to the power supply device 12, and controls the magnitude of the voltage or current to be applied, based on the detection signal input from the detection device 14.
  • FIG. 4 shows a block diagram illustrating an example of the hardware configuration of the control device 10.
  • the control device 10 includes a processor 40 such as a CPU (Central Processing Unit), a memory 42, an I/F (Interface) unit 43, a storage unit 44, a display 46, a timer 47, and an input device 48.
  • the processor 40, memory 42, I/F unit 43, storage unit 44, display 46, timer 47, and input device 48 are connected via a bus 49 such as a system bus or control bus so that various information can be exchanged between them.
  • a bus 49 such as a system bus or control bus
  • the processor 40 reads various programs, including the control program 45, stored in the storage unit 44 into the memory 42, and executes processing according to the read programs. In this way, the processor 40 controls electrophoresis.
  • the memory 42 is a work memory for the processor 40 to execute processing.
  • the control program 45 executed by the processor 40 is stored in the memory unit 44.
  • Specific examples of the memory unit 44 include a HDD (Hard Disk Drive) and an SSD (Solid State Drive).
  • the I/F unit 43 communicates various information with the power supply device 12 and the detection device 14 via wireless or wired communication.
  • the display 46 and the input device 48 function as a user interface.
  • the display 46 provides the user with various information related to the analysis of the sample.
  • the display 46 is not particularly limited, and examples of the display 46 include an LCD monitor and an LED (Light Emitting Diode) monitor.
  • the input device 48 is operated by the user to input various instructions related to the projection of the projected image.
  • the input device 48 is not particularly limited, and examples of the input device 48 include a keyboard, a touch pen, and a mouse.
  • the control device 10 employs a touch panel display that integrates the display 46 and the input device 48.
  • Timer 47 is a timer for measuring time.
  • FIG. 5 shows a functional block diagram illustrating an example of a configuration related to the functions of the control device 10 of this exemplary embodiment.
  • the control device 10 includes a detection unit 50, a timing control unit 52, and a voltage control unit 54.
  • the processor 40 executes a control program 45 stored in the memory unit 44, whereby the processor 40 functions as the detection unit 50, the timing control unit 52, and the voltage control unit 54.
  • the detection unit 50 of this exemplary embodiment has a function of monitoring the potential of the branch point 26 and detecting whether the concentrated layer has reached the branch point 26 based on the potential of the branch point 26.
  • the detection unit 50 of this exemplary embodiment controls the current value of the second branch flow path 24-2 to 0 ⁇ A by the power supply device 12. This makes the voltage value of the second flow path end 25-2 and the voltage value of the branch point 26 equal. Therefore, the detection unit 50 monitors the potential of the branch point 26 by monitoring the potential of the second flow path end 25-2 .
  • the detection unit 50 monitors the potential of the branch point 26 and derives the time differential value dV of the potential. Furthermore, the detection unit 50 compares the differential value dV with the dV threshold to detect whether or not the differential value dV has exceeded the dV threshold, and outputs a detection signal representing the detection result to the timing control unit 52.
  • the detection unit 50 of the present exemplary embodiment is an example of a detection device of the present disclosure.
  • the timing control unit 52 controls the timing of switching from isotachophoresis to capillary electrophoresis based on the detection signal input from the detection unit 50. Specifically, the timing control unit 52 determines whether the concentrated layer has reached the switching position 28 based on the detection signal, and if it determines that the concentrated layer has reached the switching position, outputs a timing detection signal indicating that to the voltage control unit 54.
  • the voltage control unit 54 outputs a control signal to the power supply unit 12 in response to the timing detection signal to switch from applying a voltage for isotachophoresis to applying a voltage for capillary electrophoresis.
  • FIG. 7 shows a flow chart illustrating an example of the flow of control processing by the processor 40 of the control device 10 of this exemplary embodiment.
  • the processor 40 of this exemplary embodiment receives an instruction to start electrophoresis, it executes the control processing shown in FIG. 7.
  • the control device 10 starts isotachophoresis and starts monitoring the potential of the branch point 26.
  • the voltage control unit 54 outputs a control signal to the power supply device 12 so as to apply a voltage for isotachophoresis to the first flow path end 25-1 and the third flow path end 25-3 of the micro-flow path chip 20.
  • the detection unit 50 also starts monitoring the potential of the branch point 26. Specifically, as described above, the detection unit 50 monitors the potential of the branch point 26 by controlling the current value of the second branch flow path 24-2 to 0 ⁇ A by the power supply device 12 and monitoring the potential of the second flow path end 25-2 .
  • the timing control unit 52 also starts counting the timer 47.
  • the control device 10 determines whether the concentrated layer has reached the branch point 26. Specifically, as described above, the detection unit 50 compares the time derivative dV of the potential at the branch point 26 with the dV threshold, and determines that the concentrated layer has reached the branch point 26 at the timing T at which the derivative dV exceeds the dV threshold. Until the concentrated layer reaches the branch point 26, the determination in step S12 is negative. On the other hand, when the concentrated layer reaches the branch point 26, the determination in step S12 is positive, and the process proceeds to step S14.
  • step S14 the timing control unit 52 of the control device 10 derives the moving speed of the concentrated layer.
  • the method by which the timing control unit 52 derives the moving speed of the concentrated layer is not particularly limited.
  • a reference time from the start of isotachophoresis until the concentrated layer reaches the branch point 26 (hereinafter referred to as the reference arrival time) is determined in advance.
  • the timing control unit 52 refers to the timer 47 and specifies the time required for the concentrated layer to reach the branch point 26 after the start of isotachophoresis in step S10 (hereinafter referred to as the actual arrival time).
  • the timing control unit 52 derives the moving speed based on the ratio or difference between the reference arrival time and the actual arrival time.
  • the moving speed of the concentrated layer derived from the length of the first branch flow channel 24.1 and the actual arrival time may be used, or instead of a specific moving speed, the ratio or difference between the reference arrival time and the actual arrival time may be used as an indirect representation of the moving speed.
  • the timing control unit 52 of the control device 10 derives the time required for the concentrated layer to move from the branch point 26 to the switching position 28. Since the distance from the branch point 26 to the switching position 28 is predetermined, the time required for the concentrated layer to reach the switching position 28 from the branch point 26 to the switching position 28 (hereinafter referred to as the "movement time") is derived from the distance from the branch point 26 to the switching position 28 and the movement speed derived in the above step S14. For example, if the timing control unit 52 derives a specific movement speed, the movement time may be derived by dividing the distance from the branch point 26 to the switching position 28 by the movement speed.
  • the movement time may be derived using the reference movement time required for the concentrated layer to move from the branch point 26 to the switching position 28 and the above ratio or difference.
  • step S18 the control device 10 refers to the timer 47 and determines whether the travel time derived in step S16 has elapsed. From the time the concentrated layer passes the branch point 26 until the travel time has elapsed, the determination in step S16 is negative. On the other hand, if the travel time has elapsed, the determination in step S16 is positive, and the process proceeds to step S20.
  • step S20 the control device 10 switches from isotachophoresis to capillary electrophoresis. Specifically, the voltage control unit 54 outputs a control signal to the power supply device 12 to stop the application of the voltage for the electrophoresis device 1 between the first flow path end 25-1 and the third flow path end 25-3 . In addition, the voltage control unit 54 outputs a control signal to the power supply device 12 to apply the voltage for capillary electrophoresis between the second flow path end 25-2 and the third flow path end 25-3 . In addition, the detection unit 50 stops monitoring the potential of the branch point 26, which was started in the above step S10. In addition, the voltage control unit 54 stops the counting by the timer 47, which was started in the above step S10. When the process of step S20 is completed, the control process shown in FIG. 7 is completed.
  • control device 10 of the electrophoresis device 1 of this exemplary embodiment can derive the travel time for the concentrated layer to travel from the branch point 26 to the switching position 28, and can recognize that the concentrated layer has reached the switching position 28 based on the travel time.
  • the detection unit 50 detects whether the concentrated layer has reached the branch point 26 by monitoring the potential of the branch point 26, but the method of detecting whether the concentrated layer has reached the branch point 26 is not limited to this embodiment.
  • the embodiment shown in Variation 1 below may also be used.
  • FIG. 8 shows a configuration diagram illustrating an example of the configuration of the electrophoresis device 1 in this modification.
  • the electrophoresis device 1 in this modification differs from the electrophoresis device 1 in the above exemplary embodiment (see Fig. 2) in that the detection device 14 further includes a sensor 15-1 and a sensor 15-2 .
  • Sensor 15-1 and sensor 15-2 are sensors for detecting fluorescence, similar to sensor 15.
  • Sensor 15-1 is disposed at a position facing reference position 27 in first branch flow channel 24-1 .
  • Sensor 15-2 is disposed at a position facing branch point 26.
  • Detection device 14 outputs detection signals corresponding to the fluorescence received as detection results of sensors 15, 15-1 , and 15-2 to control device 10. Note that in this modified example, detection device 14 is an example of a detection device disclosed herein.
  • FIG. 9 shows a functional block diagram illustrating an example of the configuration related to the functions of the control device 10 of this modified example.
  • the control device 10 of this modified example differs from the control device 10 of the exemplary embodiment described above (see FIG. 5) in that it includes an acquisition unit 51 instead of a detection unit 50.
  • part of the operation of the timing control unit 52 differs from the operation of the timing control unit 52 of the exemplary embodiment described above.
  • the acquisition unit 51 acquires detection signals representing the detection results of the sensors 15 1 and 15 2 from the detection device 14.
  • the acquisition unit 51 outputs the acquired detection signals to the timing control unit 52.
  • the timing control unit 52 of this modified example controls the timing of switching from isotachophoresis to capillary electrophoresis based on the detection signal input from the acquisition unit 51. Specifically, the timing control unit 52 determines whether the concentrated layer has reached the switching position 28 based on the detection signal, and if it determines that the concentrated layer has reached the switching position, outputs a timing detection signal indicating that to the voltage control unit 54.
  • Figure 10 shows a flowchart illustrating an example of the flow of control processing by the processor 40 of the control device 10 of this modified example.
  • step S100 of Fig. 10 the control device 10 starts isotachophoresis.
  • the voltage control unit 54 outputs a control signal to the power supply device 12 so as to apply an isotachophoresis voltage to the first flow path end 25-1 and the third flow path end 25-3 of the micro-channel chip 20.
  • the control device 10 also causes the detection device 14 to start monitoring the fluorescence state of the reference position 27 and the branch point 26.
  • the acquisition unit 51 outputs a control signal to the detection device 14 to start monitoring the fluorescence state of the reference position 27.
  • the detection device 14 acquires the detection result from the sensor 15-1 and outputs a detection signal representing the detection result to the control device 10.
  • the acquisition unit 51 outputs a control signal to the detection device 14 to start monitoring the fluorescence state of the branch point 26.
  • the detection device 14 acquires the detection result from the sensor 15-2 and outputs a detection signal representing the detection result to the control device 10.
  • the acquisition unit 51 starts acquiring a detection signal representing the detection result of the sensor 15 1 and a detection signal representing the detection result of the sensor 15 2 .
  • step S102 the control device 10 determines whether or not the concentrated layer has reached the reference position 27. Specifically, the timing control unit 52 determines whether or not the concentrated layer has reached the reference position 27 based on a detection signal representing the detection result of the sensor 15-1 . The determination in step S102 remains negative until the concentrated layer reaches the reference position 27. On the other hand, when the concentrated layer reaches the reference position 27, the determination in step S102 remains positive, and the process proceeds to step S104.
  • step S104 the timing control unit 52 of the control device 10 causes the timer 47 to start counting.
  • step S106 the control device 10 determines whether or not the concentrated layer has reached the branch point 26. Specifically, the timing control unit 52 determines whether or not the concentrated layer has reached the branch point 26 based on a detection signal representing the detection result of the sensor 152. The determination in step S106 remains negative until the concentrated layer reaches the branch point 26. On the other hand, when the concentrated layer reaches the branch point 26, the determination in step S106 remains positive, and the process proceeds to step S108.
  • step S108 the timing control unit 52 of the control device 10 derives the moving speed of the enrichment layer.
  • the method by which the timing control unit 52 derives the moving speed of the enrichment layer is not particularly limited.
  • the timing control unit 52 may derive the moving speed of the enrichment layer based on the time required for the enrichment layer to move from the reference position 27 to the branch point 26 and the distance (known) from the reference position 27 to the branch point 26, with reference to the timer 47.
  • the timing control unit 52 may derive the moving speed based on the ratio or difference between the actual time required for the enrichment layer to actually move from the reference position 27 to the branch point 26 and the reference time with reference to the timer 47. In this case, instead of a specific moving speed, the ratio or difference between the actual time and the reference time may be used to indirectly represent the moving speed.
  • the timing control unit 52 of the control device 10 derives the time required for the concentrated layer to move from the branch point 26 to the switching position 28. Since the distance from the branch point 26 to the switching position 28 is predetermined, the movement time required for the concentrated layer to reach the switching position 28 from the branch point 26 to the switching position 28 is derived from the distance from the branch point 26 to the switching position 28 and the movement speed derived in the above step S108. For example, if the timing control unit 52 derives a specific movement speed, the movement time may be derived by dividing the distance from the branch point 26 to the switching position 28 by the movement speed.
  • the timing control unit 52 derives the ratio or difference between the actual time and the reference time as an indirect representation of the movement speed
  • the movement time may be derived using the reference time required for the concentrated layer to move from the branch point 26 to the switching position 28 and the above ratio or difference.
  • step S112 the control device 10 determines whether the travel time derived in step S110 has elapsed. From the time the concentrated layer passes through the branch point 26 until the travel time has elapsed, the determination in step S112 is negative. On the other hand, if the travel time has elapsed, the determination in step S112 is positive, and the process proceeds to step S114.
  • step S114 the control device 10 switches from isotachophoresis to capillary electrophoresis.
  • the voltage control unit 54 outputs a control signal to the power supply device 12 to stop the application of the voltage for the electrophoresis device 1 between the first flow path end 25-1 and the third flow path end 25-3 .
  • the voltage control unit 54 outputs a control signal to the power supply device 12 to stop the application of the voltage for capillary electrophoresis between the second flow path end 25-2 and the third flow path end 25-3 .
  • the acquisition unit 51 outputs a control signal to the detection device 14 to stop the monitoring of the fluorescence state of the reference position 27 and the branch point 26, which was started in the above step S100.
  • step S114 the control processing shown in FIG. 10 is finished.
  • control device 10 of the electrophoresis device 1 of this modified example can also derive the travel time for the concentrated layer to travel from the branch point 26 to the switching position 28, and can recognize that the concentrated layer has reached the switching position 28 based on the travel time.
  • a reference position 27 is provided, but a configuration may be adopted in which the reference position 27 is not provided. In this case, the time from when the voltage for isotachophoresis is applied until the concentrated layer reaches the branch point 26 can be used.
  • branch point 26 is monitored has been described, but the monitoring is not limited to the branch point 26, and a form in which a position shifted upstream or downstream from the branch point 26 may also be used.
  • Second Exemplary Embodiment 11 shows a configuration diagram of an example of the electrophoresis device 1 of this exemplary embodiment.
  • the electrophoresis device 1 of this exemplary embodiment is different in that the detection device 14 (see FIG. 8) of the first modified example has sensors 15, 15 1 , 15 2 , but instead the detection device 14 has sensors 15, 15 3.
  • the sensor 15 3 is disposed at a position facing the switching position 28 in the third branch flow channel 24 3.
  • the detection device 14 outputs a detection signal corresponding to the fluorescence received as a detection result of the sensors 15, 15 3 to the control device 10. That is, in the electrophoresis device 1 of this exemplary embodiment, the sensor 15 3 directly monitors whether the concentrated layer has reached the switching position 28.
  • FIG. 12 shows a flowchart illustrating an example of the flow of control processing by the processor 40 of the control device 10 of this exemplary embodiment.
  • the control device 10 starts isotachophoresis. Specifically, the voltage control unit 54 outputs a control signal to the power supply device 12 to apply a voltage for isotachophoresis to the first flow path end 25-1 and the third flow path end 25-3 of the micro-channel chip 20.
  • the control device 10 also causes the detection device 14 to start monitoring the fluorescence state at the switching position 28. Specifically, the acquisition unit 51 outputs a control signal to the detection device 14 to start monitoring the fluorescence state at the switching position 28. As a result, the detection device 14 acquires the detection result from the sensor 15-3 , and outputs a detection signal representing the detection result to the control device 10.
  • step S152 the timing control unit 52 determines whether or not the concentrated layer has reached the switching position 28. Specifically, when the detection signal output from the sensor 153 indicates that the concentrated layer has reached the switching position 28, the timing control unit 52 determines that the concentrated layer has reached the switching position 28. The determination in step S152 remains negative until the concentrated layer reaches the switching position 28. On the other hand, when the concentrated layer has reached the switching position 28, the determination in step S152 remains positive, and the process proceeds to step S154.
  • step S154 the control device 10 switches from isotachophoresis to capillary electrophoresis. Specifically, the voltage control unit 54 outputs a control signal to the power supply device 12 to stop the application of the voltage for the electrophoresis device 1 between the first flow path end 25-1 and the third flow path end 25-3 . The voltage control unit 54 also outputs a control signal to the power supply device 12 to stop the application of the voltage for capillary electrophoresis between the second flow path end 25-2 and the third flow path end 25-3 . The acquisition unit 51 also outputs a control signal to the detection device 14 to stop the monitoring of the fluorescence state at the switching position 28, which was started in the above step S150. As a result, in the detection device 14, the detection of the fluorescence state at the switching position 28 by the sensor 15-3 is stopped. When the processing of step S154 is completed, the control processing shown in FIG. 12 is completed.
  • control device 10 of the electrophoresis device 1 of this exemplary embodiment can directly monitor the switching position 28 using the sensor 153 , thereby making it possible to recognize that the concentrated layer has reached the switching position 28.
  • control device 10 of the electrophoresis device 1 of each of the above embodiments after a concentrated layer of the sample is formed by isotachophoresis, when the concentrated layer reaches the switching position 28 in the third branch flow path 24-3 , application of voltages to the first flow path end 22-1 and the third flow path end 22-3 is stopped, and a voltage for capillary electrophoresis is applied to the second flow path end 22-2 and the third flow path end 22-3 .
  • the timing for switching from isotachophoresis to capillary electrophoresis is set to a certain time after the concentrated layer reaches branch point 26, if the moving speed of the concentrated layer changes for each measurement, the actual position of the concentrated layer at the time of switching will also change for each measurement. For example, if the length or width of the channel of microchannel chip 20 or the composition of the reagent varies from one to another, the actual position of the concentrated layer at the time of switching will change. If the position of the concentrated layer changes, the electrical characteristics of the channel will change due to the mixing of components behind the concentrated layer, which may affect the analytical performance of the sample.
  • the distance from the branch point 26 to the switching position 28 can be kept constant, so the state of the concentrated layer when switching to capillary electrophoresis can be kept constant regardless of individual differences in the microchannel chip 20 or reagents. This can improve the analytical performance of the sample.
  • the detection unit 50 detects the position of the concentrated layer based on a change in potential, and in the first modified example, the detection device 14 optically detects the position of the concentrated layer.
  • the method of detecting the position of the concentrated layer is not limited to these forms.
  • a method of magnetically detecting the position of the concentrated layer may be used.
  • a method of detecting the position of the concentrated layer may be used in which an optical camera image of the microchannel chip 20 captured by an optical camera is used to detect the position of the concentrated layer.
  • the detection device 14 is not limited to a form equipped with a sensor 15 that optically detects immune complexes, and may be of a form appropriate to the sample, etc.
  • the sensor 15 may be a sensor that magnetically detects immune complexes.
  • the various processors shown below can be used as the hardware structure of the processing units that execute various processes, such as the detection unit 50, acquisition unit 51, timing control unit 52, and voltage control unit 54.
  • the various processors include a CPU, which is a general-purpose processor that executes software (programs) and functions as various processing units, as well as programmable logic devices (PLDs), which are processors whose circuit configuration can be changed after manufacture, such as FPGAs (Field Programmable Gate Arrays), and dedicated electrical circuits, such as ASICs (Application Specific Integrated Circuits), which are processors with a circuit configuration designed specifically to execute specific processes.
  • a CPU which is a general-purpose processor that executes software (programs) and functions as various processing units, as well as programmable logic devices (PLDs), which are processors whose circuit configuration can be changed after manufacture, such as FPGAs (Field Programmable Gate Arrays), and dedicated electrical circuits, such as ASICs (Application Specific Integrated Circuits), which are processors
  • a single processing unit may be configured with one of these various processors, or may be configured with a combination of two or more processors of the same or different types (e.g., a combination of multiple FPGAs, or a combination of a CPU and an FPGA). Also, multiple processing units may be configured with a single processor.
  • Examples of configuring multiple processing units with a single processor include, first, a form in which one processor is configured with a combination of one or more CPUs and software, as typified by client and server computers, and this processor functions as multiple processing units. Secondly, a form in which a processor is used to realize the functions of the entire system, including multiple processing units, with a single IC (Integrated Circuit) chip, as typified by System On Chip (SoC). In this way, the various processing units are configured as a hardware structure using one or more of the various processors listed above.
  • SoC System On Chip
  • the hardware structure of these various processors can be an electrical circuit that combines circuit elements such as semiconductor elements.
  • control program 45 is described as being pre-stored (installed) in the storage unit 44, but this is not limiting.
  • the control program 45 may be provided in a form recorded on a recording medium such as a CD-ROM (Compact Disc Read Only Memory), a DVD-ROM (Digital Versatile Disc Read Only Memory), or a USB (Universal Serial Bus) memory.
  • the control program 45 may also be in a form downloaded from an external device via a network.
  • the program (program product) described in the exemplary embodiments may be provided on a recording medium, or may be distributed from an external computer.
  • An electrophoresis device using a micro-channel chip including at least a first branch channel connected to a first channel end, a second branch channel connected to a second channel end, and a third branch channel connected to a third channel end, which are branched at a branch point, and in which a channel through which a sample moves by electrophoresis is formed by applying a voltage to a pair of channel ends selected from the first channel end, the second channel end, and the third channel end, At least one processor; The processor, applying a first voltage for isotachophoresis to the first flow path end and the third flow path end, and moving the sample in the first branch flow path toward the third flow path end by isotachophoresis to form a concentrated layer of the sample; when the concentrated layer reaches a predetermined first position in the third branch flow path, application of the first voltage to the first flow path end and the third flow path end is stopped, and a second voltage for capillary electrophoresis is applied to the second flow path end and the third
  • the processor a detection device that detects that the concentrated layer has reached a second position that is set closer to the first flow path end than the first position in a path from the first flow path end to the third flow path end via the branch point, the path along which the sample moves by the isotachophoresis, and obtains a detection signal indicating that the concentrated layer has reached the second position; Deriving a moving speed of the concentrated layer passing through the second position based on the detection signal; Derive a travel time required for the concentrated layer to travel from the second position to the first position based on the travel speed; 2.
  • control is performed to stop application of the first voltage to the first flow path end and the third flow path end, and to apply the second voltage to the second flow path end and the third flow path end.
  • the processor Obtaining a detection result from a detection device that detects the position of the concentrated layer; 2.

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Abstract

Le dispositif d'électrophorèse selon la présente invention comporte un processeur mettant en oeuvre une commande qui applique une première tension d'isotachophorèse à une première extrémité de passage d'écoulement et une troisième extrémité de passage d'écoulement, déplace un échantillon dans un premier passage d'écoulement de ramification vers la troisième extrémité de passage d'écoulement par isotachophorèse et forme ainsi une couche concentrée de l'échantillon, et, lorsque la couche concentrée a atteint une première position prédéfinie sur un troisième passage d'écoulement de ramification, interrompt l'application de la première tension à la première extrémité de passage d'écoulement et à la troisième extrémité de passage d'écoulement, applique une seconde tension d'électrophorèse capillaire à une deuxième extrémité de passage d'écoulement et à la troisième extrémité de passage d'écoulement, et sépare un composant spécifique de la couche concentrée par électrophorèse capillaire.
PCT/JP2023/029825 2022-09-30 2023-08-18 Dispositif d'électrophorèse, procédé de commande pour dispositif d'électrophorèse et programme de commande pour dispositif d'électrophorèse WO2024070314A1 (fr)

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JPH07128286A (ja) * 1993-06-08 1995-05-19 Hewlett Packard Co <Hp> キャピラリ電気泳動システムと試料成分の分離方法
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