WO2021166210A1 - 電気泳動装置 - Google Patents

電気泳動装置 Download PDF

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Publication number
WO2021166210A1
WO2021166210A1 PCT/JP2020/006990 JP2020006990W WO2021166210A1 WO 2021166210 A1 WO2021166210 A1 WO 2021166210A1 JP 2020006990 W JP2020006990 W JP 2020006990W WO 2021166210 A1 WO2021166210 A1 WO 2021166210A1
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WIPO (PCT)
Prior art keywords
electrophoresis
current value
abnormality
flow path
electrode
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Ceased
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PCT/JP2020/006990
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English (en)
French (fr)
Japanese (ja)
Inventor
和希 野田
克洋 有留
剛 大浦
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Hitachi High Tech Corp
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Hitachi High Tech Corp
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Priority to US17/759,910 priority Critical patent/US12345994B2/en
Priority to JP2022501545A priority patent/JP7341308B2/ja
Priority to PCT/JP2020/006990 priority patent/WO2021166210A1/ja
Publication of WO2021166210A1 publication Critical patent/WO2021166210A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • G—PHYSICS
    • G02—OPTICS
    • G02F—OPTICAL DEVICES OR ARRANGEMENTS FOR THE CONTROL OF LIGHT BY MODIFICATION OF THE OPTICAL PROPERTIES OF THE MEDIA OF THE ELEMENTS INVOLVED THEREIN; NON-LINEAR OPTICS; FREQUENCY-CHANGING OF LIGHT; OPTICAL LOGIC ELEMENTS; OPTICAL ANALOGUE/DIGITAL CONVERTERS
    • G02F1/00—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics
    • G02F1/01—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour 
    • G02F1/165—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field
    • G02F1/166—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field characterised by the electro-optical or magneto-optical effect
    • G02F1/167—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field characterised by the electro-optical or magneto-optical effect by electrophoresis
    • 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
    • G01N27/44704—Details; Accessories
    • G01N27/44713—Particularly adapted electric power supply
    • G—PHYSICS
    • G02—OPTICS
    • G02F—OPTICAL DEVICES OR ARRANGEMENTS FOR THE CONTROL OF LIGHT BY MODIFICATION OF THE OPTICAL PROPERTIES OF THE MEDIA OF THE ELEMENTS INVOLVED THEREIN; NON-LINEAR OPTICS; FREQUENCY-CHANGING OF LIGHT; OPTICAL LOGIC ELEMENTS; OPTICAL ANALOGUE/DIGITAL CONVERTERS
    • G02F1/00—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics
    • G02F1/01—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour 
    • G02F1/165—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field
    • G02F1/1675—Constructional details
    • G02F1/1676—Electrodes
    • G—PHYSICS
    • G02—OPTICS
    • G02F—OPTICAL DEVICES OR ARRANGEMENTS FOR THE CONTROL OF LIGHT BY MODIFICATION OF THE OPTICAL PROPERTIES OF THE MEDIA OF THE ELEMENTS INVOLVED THEREIN; NON-LINEAR OPTICS; FREQUENCY-CHANGING OF LIGHT; OPTICAL LOGIC ELEMENTS; OPTICAL ANALOGUE/DIGITAL CONVERTERS
    • G02F1/00—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics
    • G02F1/01—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour 
    • G02F1/165—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field
    • G02F1/1675—Constructional details
    • G02F1/1679—Gaskets; Spacers; Sealing of cells; Filling or closing of cells
    • G—PHYSICS
    • G02—OPTICS
    • G02F—OPTICAL DEVICES OR ARRANGEMENTS FOR THE CONTROL OF LIGHT BY MODIFICATION OF THE OPTICAL PROPERTIES OF THE MEDIA OF THE ELEMENTS INVOLVED THEREIN; NON-LINEAR OPTICS; FREQUENCY-CHANGING OF LIGHT; OPTICAL LOGIC ELEMENTS; OPTICAL ANALOGUE/DIGITAL CONVERTERS
    • G02F1/00—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics
    • G02F1/01—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour 
    • G02F1/165—Devices or arrangements for the control of the intensity, colour, phase, polarisation or direction of light arriving from an independent light source, e.g. switching, gating or modulating; Non-linear optics for the control of the intensity, phase, polarisation or colour  based on translational movement of particles in a fluid under the influence of an applied field
    • G02F1/1685—Operation of cells; Circuit arrangements affecting the entire cell

Definitions

  • the present invention relates to an electrophoresis apparatus, and particularly to anomaly detection in a flow path.
  • Capillary electrophoresis equipment is widely used as a technique for separating and analyzing many biological samples including deoxyribonucleic acid (DNA). Since the capillary electrophoresis apparatus can perform electrophoresis using a high voltage, high-speed and high-resolution sample separation is realized.
  • Patent Document 1 in the confirmation of the state of the energizing circuit performed prior to electrophoresis, only the detection of an abnormality at the time of voltage rise was made, and the change in the state of foreign matter such as air bubbles was not taken into consideration.
  • An object of the present invention is to improve the accuracy of abnormality detection in view of the above problems.
  • the electrophoresis apparatus of the present invention includes a flow path filled with an electrophoresis medium, a first electrode provided on the cathode side of the flow path, and a second electrode provided on the anode side of the flow path.
  • an electrophoresis apparatus having a power source for applying a voltage between the first electrode and the second electrode, a pump for sending the migration medium to the flow path, and a control unit, the control unit. Is to apply a voltage to the first electrode and the second electrode prior to the filling step of filling the flow path with the migration medium, the electrophoresis step of performing the electrophoresis of the sample, and the electrophoresis of the sample.
  • Controls the analysis workflow including the abnormality detection step of determining the state of the current-carrying path based on the current value flowing through the current-carrying path including the electrophoresis medium filled in the flow path, and the control unit controls the control unit.
  • the abnormality detection step is characterized in that a voltage is applied to the current-carrying path for 20 seconds or longer.
  • the electrophoresis apparatus of the present invention can improve the accuracy of abnormality detection.
  • the schematic diagram of the electrophoresis apparatus of Example 1 The high voltage power supply circuit diagram which shows the voltage control mechanism of an electrophoresis apparatus. The figure which shows the change of the current value by the application time. The figure which shows the change of the current value by the applied voltage.
  • the analysis workflow diagram of Example 1 The schematic diagram of the electrophoretic apparatus provided with the 3rd electrode of Example 3. The analysis workflow diagram of Example 2.
  • FIG. 1 is a schematic view of an electrophoresis apparatus according to this embodiment.
  • the electrophoresis apparatus 100 includes a capillary array 102, an irradiation detection unit 104 that optically detects a sample separated by electrophoresis, and a pump unit 106 that feeds an electrophoresis medium.
  • the capillary array 102 includes a single or a plurality of capillaries 108, a load header 110, and a capillary head 112.
  • the capillary 108 is composed of a glass tube having an inner diameter of several tens to several hundreds of microns and an outer diameter of several hundreds of microns, and the surface of the capillary 108 is coated with a polyimide resin in order to improve the strength.
  • the detection position 114 of the capillary 108 for reading the sample information by the irradiation detection unit 104 has the polyimide resin coating removed so that the internal light emission leaks to the outside.
  • the irradiation detection unit 104 has a light source 116 and an optical detector 118, and the excitation light from the light source 116 is applied to the sample at the detection position 114, and light having a wavelength depending on the sample is emitted. The emitted light is detected by the optical detector 118.
  • the load header 110 is provided on the cathode side of the capillary 108.
  • a hollow electrode 120 is attached to the load header 110, and the load header 110 and the hollow electrode 120 are integrated.
  • the cathode end of the capillary 108 passes through the hollow electrode 120 and is fixed in a state of protruding from the hollow electrode 120.
  • the hollow electrode 120 is conductive with the high-voltage power supply 122, and when a voltage is applied such as electrophoresis or sample introduction, the hollow electrode 120 functions as a cathode electrode.
  • the electrophoresis apparatus 100 is provided with a first ammeter 124 and a second ammeter 126 for detecting the current in the current-carrying path when a voltage is applied by the high-voltage power supply 122.
  • a cathode buffer container 129 containing a buffer 128, a cleaning container 131 containing pure water 130, a waste liquid container 132, and a sample container 135 containing a solution 134 containing a sample are provided at the cathode end of the capillary 108. It is conveyed by a transfer machine 136 that can move to three axes.
  • the capillary head 112 is a member that is attached to and detached from the pump unit 106 with pressure resistance confidentiality.
  • the capillary head 112 bundles the capillary anode ends into one when there are a plurality of capillary 108s.
  • the inside of the capillary 108 is filled with the electrophoresis medium 137 by the pump unit 106.
  • the electrophoresis medium 137 gives a difference in migration rate to the sample during electrophoresis.
  • the pump unit 106 is mainly composed of a block 138 in which a flow path is formed and a pump 140.
  • the block 138 is a connection portion for communicating the capillary array 102, the electrophoresis medium container 142 containing the electrophoresis medium 137, and the anode buffer container 144, and communicates the capillary array 102, the electrophoresis medium container 142, and the anode buffer container 144. A flow path for this is formed.
  • a connecting pipe 146 connecting the anode buffer container 144 and the block 138 is connected to the block 138.
  • the connecting pipe 146 is provided with a valve 148.
  • the migration medium 137 contained in the migration medium container 142 with the valve 148 open is pumped by the pump 140, the migration medium 137 passes through the flow path formed in the block 138 and the connecting tube 146, and the anode. The liquid is sent toward the buffer container 144. As a result, the flow path in the block 138 is filled with the electrophoresis medium 137.
  • the running medium 137 is sent in a state where the valve 148 is closed, the running medium 137 is sent to the capillary array 102, and the inside of the capillary 108 can be filled with the running medium 137.
  • a check valve 152 is provided between the block 138 and the migration medium container 142 so that the migration medium 137 sucked from the migration medium container 142 does not flow back.
  • the connecting tube 146 and the electrode (GND) 156 are inserted into the anode buffer container 144 so as to be immersed in the buffer 154 in the anode buffer container 144. If a voltage is applied while the connecting pipe 146 is not immersed in the buffer 154, a discharge may occur. Further, it is desirable that the positions of the liquid levels of the buffer 154 and the buffer 128 housed in the anode buffer container 144 and the cathode buffer container 129 are about the same. This is to prevent the buffer 154 (128) from flowing into the capillary 108 due to the difference in pressure due to the difference in the height of the liquid level. Further, in this embodiment, the pump 140 is used to send the running medium 137, but the user may manually send the running medium 137 into the flow path using a syringe or the like.
  • the electrophoresis apparatus 100 has a constant temperature bath 158 for keeping the temperature of the capillary array 102 constant during the electrophoresis of the sample, and the capillary array 102 is arranged in the constant temperature bath 158. Further, the electrophoresis apparatus 100 is used in a state of being connected to the computer 160 by a communication cable 162 or the like.
  • the computer 160 is connected to the microcomputer 164 of the electrophoresis apparatus 100.
  • the computer 160 can control the operation of the electrophoresis apparatus 100 and the functions possessed by the computer 160, and can exchange data detected by the electrophoresis apparatus 100.
  • the computer 160 and the microcomputer 164 will be described in detail as control units, but one control unit may play the roles of the computer 160 and the microcomputer 164.
  • FIG. 2 is a high-voltage power supply circuit diagram showing the voltage control mechanism of this device. The voltage control mechanism will be described below with reference to FIG.
  • the voltage control mechanism includes a microcomputer 164, a controller 166, a high voltage power supply 122, a first ammeter 124, and a second ammeter 126. Based on the control of the controller 166, the output of the high-voltage power supply 122 and the voltage applied to the energization path are performed.
  • the energization path is a hollow electrode 120, a buffer 128 filled in the cathode buffer container 129, an electrophoresis path, a buffer 154 filled in the anode buffer container 144, and an electrode (GND) 156.
  • the electrophoresis path is an electrophoresis medium 137 filled in a capillary 108, a flow path formed in a block 138, and a connecting tube 146 (see FIG. 1).
  • the high-voltage power supply 122 is conducting with the hollow electrode 120 via the first ammeter 124 and with the electrode (GND) 156 via the second ammeter 126.
  • the second ammeter 126 is connected to the microcomputer 164.
  • a voltage is applied by the high voltage power supply 122, an electric field is generated between the hollow electrode 120 and the electrode (GND) 156. Due to this electric field, the negatively charged sample moves from the cathode side of the capillary 108 to the anode side of the capillary 108.
  • the first ammeter 124 detects the current flowing from the high-voltage power supply 122 to the hollow electrode 120, and transmits the current value to the microcomputer 164.
  • the second ammeter 126 detects the current flowing from the electrode (GND) 156 to GND, and transmits the current value to the microcomputer 164. In the abnormality detection described later, the value of the second ammeter 126 is used as the current value flowing through the energizing path.
  • a medium having a relatively large resistance as compared with metal such as a buffer 154 (168) and an electrophoresis medium 137, is interposed between the first ammeter 124 and the second ammeter 126. Further, there are many connections such as a block 138 and a capillary array 102 between the first ammeter 124 and the second ammeter 126.
  • the area between the first ammeter 124 and the second ammeter 126 is a portion where noise is likely to occur in the measured current value.
  • the numerical value indicated by the second ammeter 126 is unlikely to include noise.
  • the second ammeter 126 detects the net current value flowing through the electrophoresis path.
  • the microcomputer 164 reads the current values from the first ammeter 124 and the second ammeter 126 and performs the calculation. Then, a command is sent to the controller 166 to control the high voltage power supply 122 to each state such as high voltage application, low voltage application, and voltage cutoff.
  • a voltage is applied between the hollow electrode 120 and the electrode (GND) 156 and the current flowing through the current path is compared with the threshold value prior to the electrophoresis of the sample.
  • abnormality detection is performed to determine the state of the current-carrying path.
  • the resistance of the foreign matter makes it difficult for the current to flow, and the current value becomes smaller than when there is no foreign matter. Therefore, the state of the energizing path can be determined by comparing the current value and the threshold value.
  • Patent Document 1 only the rising time of the current is conscious, and only the detection of foreign matter that can be detected at the rising of the current is considered.
  • foreign matter such as minute bubbles that cannot be detected as foreign matter at the time of starting the current may expand due to Joule heat applied by the voltage after the abnormality detection step, blocking the electrophoresis path and causing discharge.
  • a voltage larger than that in the abnormality detection step is applied. Therefore, the voltage application after the abnormality detection step is more likely to cause a failure of the pump unit 106 or damage to the capillary 108 than the voltage application in the abnormality detection step, and it is preferable to detect foreign matter in the abnormality detection step.
  • the analysis operation is stopped. If the detection of foreign matter is delayed, the return work will increase accordingly, and the time required for analysis will increase.
  • FIG. 3 is a graph showing the change in the current value with respect to the voltage application time when a voltage of 2 kilovolts is applied between the hollow electrode 120 and the electrode (GND) 156 in a state where air bubbles are mixed.
  • the current value is measured by intentionally mixing bubbles in the electrophoresis path. If it is lower than the predetermined threshold value as described above, it is determined that there is an abnormality.
  • the solid line data shows that bubbles can be detected when the current rises (for example, 3 seconds after the voltage is applied), and the dotted line data shows that bubbles are not detected when the current rises.
  • the application time is set to 60 seconds or less so that there is no danger of electric discharge or the like even in an abnormal state. Further, increasing the application time enhances the foreign matter detection performance, but if the application time is too long, the time required for the analysis workflow will be extended, so the above-mentioned 20 seconds or more and 60 seconds or less is preferable. be.
  • the horizontal axis is the applied voltage applied to the hollow electrode 120 and the electrode (GND) 156
  • the vertical axis is the current value flowing through the energizing path acquired by the second ammeter 126.
  • the applied voltage and the detected current value have a roughly linear relationship.
  • the applied voltage was small, the current value was low even in the absence of foreign matter, and the difference in current value depending on the presence or absence of foreign matter was small.
  • the applied voltage is 3 kilovolts or less, it can be confirmed that the detected current values overlap depending on the presence or absence of bubbles.
  • the applied voltage when the applied voltage is small, it is difficult to set the optimum threshold value, and it is difficult to identify the presence or absence of an abnormality.
  • the applied voltage when the applied voltage is large, there is a large difference in the detected current value between the state without foreign matter and the state with foreign matter, so that it becomes easy to determine whether or not there is an abnormality.
  • the applied voltage when there are bubbles decreases from 4 kilovolts to 5 kilovolts
  • the bubbles and the like are likely to expand and the detected current is further reduced. Therefore, the larger the applied voltage, the more accurately the foreign matter can be detected.
  • a voltage of 4 kilovolts or more it is preferable to apply a voltage of 4 kilovolts or more to detect an abnormality.
  • the applied voltage is made too large, there is a risk of discharge or the like. Therefore, in abnormality detection, it is preferable to apply a voltage of about 4 kilovolts to 6 kilovolts at the time of abnormality detection, in which discharge or the like is unlikely to occur even in an abnormal state such as air bubbles or foreign matter being mixed in the flow path.
  • the operator installs the cathode buffer container 129, the washing container 131, the anode buffer container 144, the electrophoresis medium container 142, the capillary array 102, and the sample container 135 in the apparatus. Since the cathode buffer container 129, the washing container 131, the anode buffer container 144, the electrophoresis medium container 142, the capillary array 102, and the sample container 135 are all replacement members, they are replaced at predetermined timings.
  • the capillary array 102 and the migration medium container 142 are replaced by using the pump unit 106 before performing the analysis workflow.
  • the flow path formed in 108 and block 138 is filled with the migration medium 137.
  • the analysis workflow will be described with reference to FIG.
  • the analysis workflow is executed upon receiving a start instruction from computer 160 (300).
  • the pump 140 first fills the flow path and connecting tube 146 formed in the block 138 with the migration medium 137 (301).
  • the capillary array 102 is filled with the migration medium 137 (302).
  • abnormality detection is performed to confirm whether or not there is an abnormality in the energizing path (303).
  • a voltage smaller than the voltage used for electrophoresis is applied as described above, and the current value flowing in the energization path at that time is measured. The presence or absence of abnormality is confirmed by comparing the measured current value with a predetermined threshold value.
  • a voltage of 4 kilovolts or more for 20 seconds or more it is preferable to apply a voltage of 4 kilovolts or more for 20 seconds or more.
  • preliminary electrophoresis is executed (304).
  • the preliminary electrophoresis is for making the state of the electrophoresis medium 137 in the capillary 108 suitable for electrophoresis.
  • a voltage of several to several tens of kilovolts is applied for several tens of minutes.
  • the current is measured during the preliminary run to detect abnormalities.
  • a sample is introduced into the capillary 108 (305).
  • the sample container 135 is transferred by the transfer machine 136, and the capillary 108 cathode end and the hollow electrode 120 are immersed in the solution 134 containing the sample.
  • the sample is introduced into the capillary 108 by applying a voltage between the hollow electrode 120 and the electrode (GND) 156 by the high voltage power supply 122. Then, the transfer machine 136 transfers the cathode buffer container 129 so that the capillary 108 cathode end and the hollow electrode 120 penetrate the buffer 128, and starts electrophoresis (306). By applying a voltage between the hollow electrode 120 and the electrode (GND) 156 by the high voltage power supply 109, the sample to be detected is separated while moving in the capillary 108. The sample is detected by the irradiation detection unit 104 as it passes through the detection position 114 of the capillary 108.
  • an abnormality is detected in the abnormality detection in step 303 or the preliminary electrophoresis in step 304, it is necessary to eliminate the abnormal state. If an abnormality is detected, it is considered that the cause of the abnormality is insufficient liquid volume in the buffer 128 (154), residual air bubbles or foreign matter in the electrophoresis path, damage to the capillary 108, etc., because there is an abnormality in the energizing path. Be done. In the past, when an abnormality was confirmed, the error was immediately notified and the operator was requested to take action. The operator visually confirmed the abnormal state and took predetermined measures according to the cause of the confirmed abnormality. Foreign matter is removed when foreign matter is found in the flow path.
  • the foreign matter is automatically removed, so that the frequency of requesting the operator to respond can be reduced and the operability can be improved. Further, since a predetermined flow is automatically performed when an abnormality is detected, it is possible to reduce individual differences in abnormality detection.
  • step 303 or step 304 when an abnormality is detected, the number of times the abnormality is detected in step 303 or step 304 is recorded in a control unit such as a computer 160 or a microcomputer 164 (307).
  • a control unit such as a computer 160 or a microcomputer 164.
  • the process returns to step 301, the flow path of the block 138 and the capillary 108 are refilled with the migration medium 137, and the abnormality is detected in step 303.
  • the current values measured in step 303 before and after the refilling of the electrophoresis medium 137 are compared (308). When the bubbles expand or the foreign matter moves, the measured current value fluctuates.
  • the change in the state of the foreign matter is confirmed by comparing the current values in step 303 before and after the refilling of the migration medium 137.
  • the change in the current value measured in step 303 before and after the refilling of the migration medium 137 is larger than a predetermined threshold value, it is determined that the abnormal state has changed.
  • the abnormal state is changed by refilling the migration medium 137, there is a high possibility that the abnormal state will be released by filling the migration medium 137. Therefore, returning to step 301, the foreign matter is attempted to be removed by refilling the migration medium 137. If the change in the current value before and after refilling the migration medium 137 in step 308 is less than the threshold value, it is considered that the abnormal state has not changed.
  • the electrophoresis medium 137 is strongly fed by increasing the liquid feeding pressure or increasing the liquid feeding speed (309). .. Specifically, by increasing the drive current of the pump 140, the force for pushing out the electrophoresis medium 137 of the pump 140 is increased. Alternatively, when the pump 140 is driven by a motor, the rotation speed of the motor is increased to feed the migration medium 137 faster. After that, the abnormality is detected by measuring the current value in the same manner as the abnormality detection in step 303 (310). If no abnormality is detected, the foreign matter has been removed, and the preliminary electrophoresis in step 304 is performed.
  • the current value measured in the previous step 303 is compared with the current value measured in step 310 (311).
  • the change in the current value is less than the threshold value, it is considered that the abnormal state has not changed, so that it is highly possible that the cause of the error is other than the foreign matter in the electrophoresis path.
  • the error is notified and the operator is requested to take action (313).
  • the process returns to step 301 to refill the migration medium 137.
  • the number of times an abnormality is detected is recorded in step 307, and a threshold value is set for the number of times an abnormality is detected in step 303 or step 304. If the number of times the abnormality recorded in step 307 is detected exceeds the threshold value, an error is displayed and the device is stopped (313).
  • a threshold value it is possible to suppress the consumption of the migration medium 137 more than necessary, and by removing the foreign matter a predetermined number of times, it is possible to show that there is a high possibility of an abnormality other than the foreign matter when an error is displayed. It is possible.
  • a threshold value is set for the number of times an abnormality is detected in step 303 or step 304, but the same analysis can be performed by recording the number of filling steps of steps 301 to 302 and setting a threshold value for the number of times. It is possible to carry out the workflow.
  • the electrophoresis apparatus 600 of this embodiment has a third electrode 602 for confirming the position where the abnormality occurs.
  • the apparatus configuration is the same as that of the first embodiment except that the third electrode 602 is provided.
  • the electrophoresis apparatus 600 provided with the third electrode 602 of this embodiment will be described with reference to FIG.
  • the flow path that communicates the migration medium container 142 and the capillary 108 is the first flow path 604, and the anode buffer container 144 in which the electrode (GND) 156 is housed.
  • the position where the second flow path 606 communicates with the capillary 108, the first flow path 604 and the second flow path 606 intersect, and the capillary head 112 is connected is referred to as the capillary connection portion 608.
  • the electrophoresis apparatus 600 of this embodiment newly includes a third electrode 602 in the first flow path 604.
  • a third electrode 602 in the first flow path 604.
  • the current value between the hollow electrode 120 and the electrode (GND) 156 and the current value between the hollow electrode 120 and the third electrode 602 are measured, and the state of the energization path is determined. to decide.
  • the abnormality detection of this embodiment is performed by using the current value between the hollow electrode 120 and the electrode (GND) 156 as the first current value and the current value between the hollow electrode 120 and the third electrode 600 as the second current value. explain.
  • the abnormality detection pattern there is no error in the first current value and the second current value (case 1), the abnormality is detected only in the first current value (case 2), and the abnormality is detected only in the second current value (case 2).
  • case 3 abnormality detection
  • case 4 abnormality detection
  • a foreign substance in the capillary 108, the capillary connecting portion 608, the second flow path 606, or the connecting pipe 116, or an error in the case of an abnormal state other than the foreign substance is detected.
  • a foreign matter in the capillary 108, the capillary connection portion 608 or the first flow path 604, or an abnormal state other than the foreign matter is detected. Therefore, when an abnormality is determined based on either the first current value or the second current value, it can be determined that the cause of the abnormality can be detected by only one of them.
  • the cause of the error is a foreign substance in the second flow path 606 or the connecting pipe 146, and the abnormality is determined only from the second current value (case 2).
  • the cause of the error is a foreign substance in the first flow path 604.
  • the block 138 and the capillary 108 were refilled with the electrophoresis medium 137.
  • the cause of the abnormality can be identified as a foreign substance in the block 138 or the connecting tube 146, the foreign substance is removed by sending the electrophoresis medium 137 to the flow path of the block 138.
  • the third electrode 602 is preferably provided at the connection portion of the migration medium container 142 of the block 138. Further, although the third electrode 602 is provided in the block 138 in FIG. 6, it may be inserted into the electrophoresis medium container 142.
  • FIG. 7 will be used to explain a part of this embodiment that is different from the analysis workflow of the first embodiment.
  • the flow when refilling the migration medium 137 is different from that in Example 1. Since the other flows are the same as those in the first embodiment, the description thereof will be omitted.
  • the abnormality detection in steps 303 and 310 the abnormality is determined based on the first current value and the second current value, and the abnormality is detected based on at least one of the first current value and the second current value. If so, it is determined that there is an abnormality. When it is determined that there is no abnormality based on the first current value and the second current value (case 1), it is determined that there is no abnormality.
  • FIG. 7 although the illustration of steps 307 to 311 is omitted for the sake of legibility, the same applies to steps 307 to 311 in FIG. 5 of the first embodiment.
  • the migration medium 137 is to be refilled in steps 307 to 311 (700)
  • the result of abnormality detection is confirmed in this embodiment (701).
  • the flow differs depending on the result of abnormality detection.
  • the cause of the abnormality is in the second flow path 606 or in the connecting pipe 146, respectively, as described above. Since it can be determined that the foreign matter is the foreign matter in the first flow path 604 or the foreign matter in the first flow path 604, the flow path formed in the block 138 is filled with the migration medium 137 (702). When the flow path of the block 138 is filled with the migration medium 137, the migration medium 137 in the migration medium container 142 passes through the first flow path 604, the capillary connection portion 608, the second flow path 606, and the connecting tube 146, and is an anode buffer.
  • the liquid is sent to the container 144, and the foreign matter is discharged to the anode buffer container 144. If an abnormality is detected in the first current value of Case 2, the cause is foreign matter in the second flow path 606 or the connecting pipe 146, and the foreign matter is located near the anode buffer container 144 to which the foreign matter is discharged. ing. Since it is sufficient to send the migration medium 137 filled with the second flow path 606 and the connecting tube 146, a smaller amount of the running medium 137 than the running medium filling in steps 301 and 702 is sent to the flow path of the block 138 ( 703). By adjusting the liquid feed amount of the migration medium 137 in Case 2 and Case 3, it is possible to further reduce the amount of the migration medium 137 used.
  • Case 4 When an abnormality is detected in the first current value and the second current value of the case 4, foreign matter in the capillary 108, foreign matter in the first flow path 604, foreign matter in the second flow path 606 or the connecting pipe 146, etc. Various cases are possible. Therefore, in Case 4, the process returns to step 302, and the flow path of the block 138 and the capillary 108 are refilled with the migration medium 137 in the same manner as in the first embodiment.
  • the state of the energizing path is confirmed before electrophoresis, and if there is an abnormality, the system for removing the mixed foreign matter and the number of times of removing the foreign matter are limited.
  • the current value and the threshold value are compared to determine the state of the current-carrying path, but the abnormal state may be determined from the slope of the current value or the like.
  • Electrophoretic apparatus 102: Capillary array, 104: Irradiation detection unit, 106: Pump unit, 108: Capillary, 110: Load header, 112: Capillary head, 114: Detection position, 116: Light source, 118: Optical detector , 120: Hollow electrode, 122: High-voltage power supply, 124: First electrophoresis meter, 126: Second electrophoretic meter, 128: Buffer, 129: Cathode buffer container, 130: Pure water, 131: Cleaning container, 132: Waste liquid container, 134: Solution containing sample 135: Sample container, 136: Conveyor, 137: Electrophoretic medium, 138: Block, 140: Pump, 142: Electrophoretic medium container, 144: Anode buffer container, 146: Connecting tube, 148: Valve , 152: Check valve, 154: Buffer, 156: Electrode (GND), 158: Constant temperature bath, 160: Computer, 162: Communication

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PCT/JP2020/006990 2020-02-21 2020-02-21 電気泳動装置 Ceased WO2021166210A1 (ja)

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WO2024246989A1 (ja) * 2023-05-26 2024-12-05 株式会社日立ハイテク 異常検知方法及び異常検知装置

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JP2007107915A (ja) * 2005-10-11 2007-04-26 Shimadzu Corp キャピラリ流路における電気泳動方法及びマイクロチップ処理装置
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JPS60119457A (ja) * 1983-11-30 1985-06-26 Shimadzu Corp 電気泳動装置
JP2003344356A (ja) * 2002-05-31 2003-12-03 Hitachi High-Technologies Corp 電気泳動装置、及び電気泳動方法
JP2007107915A (ja) * 2005-10-11 2007-04-26 Shimadzu Corp キャピラリ流路における電気泳動方法及びマイクロチップ処理装置
JP2012068234A (ja) * 2010-08-10 2012-04-05 Arkray Inc 電気泳動装置、および電気泳動装置の制御方法
JP2012093352A (ja) * 2010-09-28 2012-05-17 Arkray Inc 気泡除去方法、気泡除去装置、それを用いた分析装置、気泡除去制御プログラム、およびこのプログラムの記憶媒体

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WO2023026366A1 (ja) * 2021-08-24 2023-03-02 株式会社日立ハイテク 電気泳動支援方法
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WO2024246989A1 (ja) * 2023-05-26 2024-12-05 株式会社日立ハイテク 異常検知方法及び異常検知装置

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