WO2023140324A1 - Dispositif de collecte, microscope à conductance ionique à balayage qui en est équipé et procédé de collecte - Google Patents

Dispositif de collecte, microscope à conductance ionique à balayage qui en est équipé et procédé de collecte Download PDF

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Publication number
WO2023140324A1
WO2023140324A1 PCT/JP2023/001520 JP2023001520W WO2023140324A1 WO 2023140324 A1 WO2023140324 A1 WO 2023140324A1 JP 2023001520 W JP2023001520 W JP 2023001520W WO 2023140324 A1 WO2023140324 A1 WO 2023140324A1
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Prior art keywords
pipette
reference electrode
pseudo reference
sample
tip
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PCT/JP2023/001520
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English (en)
Japanese (ja)
Inventor
大貴 井田
明哉 熊谷
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国立大学法人東北大学
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Publication of WO2023140324A1 publication Critical patent/WO2023140324A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/26—Inoculator or sampler
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/42—Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01Q—SCANNING-PROBE TECHNIQUES OR APPARATUS; APPLICATIONS OF SCANNING-PROBE TECHNIQUES, e.g. SCANNING PROBE MICROSCOPY [SPM]
    • G01Q60/00—Particular types of SPM [Scanning Probe Microscopy] or microscopes; Essential components thereof
    • G01Q60/44—SICM [Scanning Ion-Conductance Microscopy] or apparatus therefor, e.g. SICM probes

Definitions

  • the present invention relates to a collection device, a scanning ion conductance microscope equipped with the same, and a collection method.
  • a scanning probe microscope uses a sharp tip (probe) instead of a lens, and when the probe is brought close to the sample, it acquires physical information and chemical responses generated by the interaction between the sample and the probe as signals.
  • This microscope is widely used as a microscope capable of obtaining image information of the surface of a sample by scanning the surface of the sample with a probe while feeding back the above-described signals.
  • a scanning ion conductance microscope which is one of scanning probe microscopes, uses a glass pipette electrode as a probe and utilizes changes in ion current generated between it and a pseudo-reference electrode placed in an electrolytic solution in which the sample is immersed. Scanning ion conductance microscopes are suitable for observation of soft biological samples in liquid, and are said to be promising for observation of various biological samples.
  • FIG. 9 is an explanatory diagram showing the operating principle of a scanning ion conductance microscope, in which a glass pipette 101 filled with an electrolyte solution and equipped with a first pseudo reference electrode 100 is immersed in the electrolyte solution 102.
  • the second pseudo reference electrode 103 is immersed in the electrolyte solution 102 , and the glass pipette 101 is scanned while the first pseudo reference electrode 100 and the second pseudo reference electrode 103 are connected to the current measuring device 104 and the voltage source 105 .
  • a voltage is applied between the first pseudo reference electrode 100 and the second pseudo reference electrode 103, an ion current is generated at the tip of the pipette, and the resulting ion current can be obtained.
  • the ion current at the tip of the pipette is blocked from a certain point when the glass pipette 101 is very close to the biological sample 106, and the ion current decreases.
  • a typical cell size is about several tens of ⁇ m, and the volume is on the order of pL (pL; picoliters: 10 ⁇ 12 L).
  • These cells contain higher-order structures such as organelles, and the cells function by working in concert with each other.
  • Higher-order structures are composed of nucleic acids, proteins, lipids, and the like, and are approximately ⁇ m to nm in size and fL to aL in volume (fL: femtoliter: 10 ⁇ 15 L, aL: attoliter: 10 ⁇ 18 L).
  • the inventor of the present invention has arrived at the present invention as a result of researching a technology that can control the maximum collection amount and maintain reproducibility and accuracy in nanoglass pipettes that are applied to scanning ion conductance microscopes and the like, even with pipettes that have various opening diameters, shapes, and chemical modifications.
  • the object of the recovery device of the present invention is to provide a technology that enables precise suction and ejection at a minute level, which could not be achieved with conventional voltage control technology, while using a nanoglass pipette with a tip opening diameter on the order of several tens of nm to several ⁇ m.
  • Another object of the present invention is to provide a recovery device and a recovery method capable of increasing the maximum suction volume and discharge volume or restricting the recovery volume of pipettes that can be used in the prior art, thereby enabling highly accurate liquid volume control.
  • An object of the present invention is to provide a scanning ion conductance microscope equipped with the above-described nanoglass pipette that enables precise suction and discharge at a minute level without being affected by the aperture diameter, which has not been possible in the past.
  • a recovery device comprises a pipette having a liquid suction port at its tip, a first pseudo reference electrode inserted into the pipette, a second pseudo reference electrode immersed in the electrolytic solution into which the pipette is inserted, a voltage source connected to the first pseudo reference electrode and the second pseudo reference electrode, a current measuring device, and a pump connected to the proximal end of the pipette to apply negative or positive pressure to the inside of the pipette.
  • the pump is connected to the base end of the pipette through a pipe, a joint member is incorporated in the pipe, and the first pseudo reference electrode is electrically connected to the pipe and the joint member, and is connected to a current measuring instrument through external wiring.
  • a current measuring device is incorporated in wiring connecting the voltage source, the first pseudo reference electrode, and the second pseudo reference electrode.
  • a pressure gauge is connected to the pipe connecting the pump and the pipette.
  • the inner diameter of the suction port of the pipette is on the order of nm to ⁇ m.
  • a scanning ion conductance microscope according to the present invention is characterized by comprising the collection device according to any one of (1) to (5), wherein the pipette is supported by a triaxial stage.
  • the scanning ion conductance microscope according to the present invention is characterized in that the three-axis stage has a three-axis primary stage capable of movement with a range of motion on the order of mm and an accuracy on the order of ⁇ m, and a three-axis secondary stage capable of movement with a range of motion on the order of ⁇ m and an accuracy on the order of nm.
  • the recovery method according to the present invention uses a pipette having an inlet at its tip, a first pseudo reference electrode inserted inside the pipette, a pump that applies pressure such as hydraulic pressure to the inside of the pipette, and a second pseudo reference electrode. is controlled to suck the electrolyte solution or the sample contained in the electrolyte solution into the pipette. Further, in the recovery method of the present invention, it is also possible to discharge the solution sucked into the pipette, and it is also possible to discharge an aqueous solution or a sample contained in the aqueous solution into the electrolyte solution.
  • a pipette having a suction port at its tip and an inner diameter of the suction port on the order of nm to ⁇ m.
  • a sample is immersed in the electrolyte solution, and the sample or a part of the sample is sucked with the pipette together with the electrolyte solution.
  • the collection device while using a pipette whose tip suction port diameter is on the order of nanometers, in addition to conventional voltage control technology, by applying pressure from a hydraulic pump or the like to the pipette, it is possible to aspirate and discharge a volume of liquid that could not be achieved with conventional voltage control technology alone.
  • a pipette whose tip suction port has a diameter on the order of ⁇ m it is possible to accurately control the suction and discharge of minute-level liquids, which could not be achieved with conventional techniques.
  • the tip of the pipette can be scanned with respect to the sample in the electrolyte solution by the three-axis stage while generating an ionic current by applying a voltage between the first pseudo reference electrode in the pipette and the second pseudo reference electrode immersed in the electrolyte solution.
  • an ionic current by applying a voltage between the first pseudo reference electrode in the pipette and the second pseudo reference electrode immersed in the electrolyte solution.
  • pressure is applied to the pipette, a voltage is applied between the first pseudo-reference electrode and the second pseudo-reference electrode of the pipette immersed in the electrolyte solution, and the voltage is controlled while applying pressure, so that the electrolyte solution can be sucked into the pipette. Also, by adjusting the voltage, the electrolyte solution can be discharged from the pipette. Since not only voltage control but also pressure is used, more electrolyte solution can be sucked into the pipette than before, and more minute amounts of electrolyte solution can be aspirated and collected more accurately than before.
  • FIG. 1 is a configuration diagram showing a recovery device according to a first embodiment of the present invention
  • FIG. It is an overall schematic diagram showing the relationship between the three-axis stage, the nanoglass pipette, and the sample provided in the same scanning ion conductance microscope. It is a partially enlarged view showing the relationship between the three-axis stage provided in the same scanning ion conductance microscope, the nanoglass pipette, and the sample, and is a schematic diagram of the pipette sticking into the cell.
  • 10 is a graph showing normalized current values generated according to the pipette-sample distance in the same scanning ion conductance microscope.
  • FIG. 10 is a graph of applied voltage and current value in a state in which different pressures are applied by hydraulic pressure to the nanopipette provided in the same scanning ion conductance microscope.
  • FIG. It is explanatory drawing which shows each part size of the nanopipette provided in the same scanning ion conductance microscope.
  • 4 is a graph showing the relationship between oil pressure and recovery amount when a glass pipette with an inlet diameter of 700 nm is used.
  • 4 is a graph showing the relationship between oil pressure and recovery amount when a glass pipette with an inlet diameter of 100 nm is used.
  • 4 is a graph showing the relationship between oil pressure and recovery amount when a glass pipette with an inlet diameter of 70 nm is used.
  • FIG. 4 is an explanatory diagram showing a measurement concept of a scanning ion conductance microscope;
  • a first embodiment of the present invention will be described in detail below with reference to the accompanying drawings.
  • characteristic portions are enlarged for convenience in order to make the characteristics easier to understand.
  • the glass pipette 1 has a tapered tip portion 1A, and has an inlet 1a at the tip of the tip portion 1A.
  • the diameter of the suction port 1a is, for example, about 10 nm to 1000 nm. More specifically, the diameter of the suction port 1a can be formed in the range of about 30 nm to several ⁇ m.
  • a central portion 1B and a rear end portion (base end portion) 1C of the glass pipette 1, excluding the tip portion 1A, are formed in a tubular shape having the same outer diameter as that of the raw material capillary.
  • the glass pipette 1 is supported vertically downward by a support member (not shown) in a state in which the tip portion 1A is immersed in an electrolytic solution S contained in a container 7 such as a petri dish.
  • the diameter of the suction port of the glass pipette 1 described above is merely an example, and is not limited to the range described above.
  • a capillary tube made of borosilicate glass (borosilicate glass) or quartz glass is pulled by a laser puller to reduce the inner diameter of the tip to the diameter described above.
  • the glass pipette 1 may be made of other materials such as resins and ceramics as long as the material can form the above diameter.
  • a tubular metal joint member (screw joint) 11 is connected to the rear end (base end) of the glass pipette 1 via a first pipe (first pressure pipe) 10 such as a flexible resin pipe.
  • the joint member 11 has a first connection portion (first tube joint) 12 formed on one end thereof, a second connection portion (second tube joint) 13 formed on the other end side, and a first pipe 10 is connected to the first connection portion 12.
  • a pump 6 is connected to a second connecting portion 13 of the joint member 11 via a second pipe (second pressure pipe) 14 such as a flexible resin pipe.
  • a T-shaped joint 15 is incorporated in the middle of the second pipe 14 , and a pressure measuring device (pressure gauge) 17 is connected to the T-shaped joint 15 via a third pipe (third pressure pipe) 16 .
  • a pressure measuring device 17 makes it possible to measure the pressure exerted by the pump 6 inside the glass pipette 1 .
  • the first pseudo reference electrode 2 has its lower end located in the central portion 1B of the glass pipette 1 and is arranged so as to pass through the first pipe 10. The upper end of the first pseudo reference electrode 2 is electrically connected to the inner wall of the joint member 11.
  • a first wiring 18 connected to the voltage source 5 is connected to the peripheral wall of the joint member 11 .
  • a (micro)current measuring device 19 is incorporated in the first wiring 18 between the voltage source 5 and the joint member 11 .
  • a second wiring 20 is connected to the pole of the voltage source 5 opposite to the pole to which the first wiring 18 is connected, the second wiring 20 is connected to the second pseudo reference electrode 3, and the lower end side of the second pseudo reference electrode 3 is immersed in the electrolyte solution S.
  • a necessary voltage can be applied between the first pseudo reference electrode 2 and the second pseudo reference electrode 3 from the voltage source 5 .
  • An Ag/AgTPBCl electrode can be used for the first pseudo reference electrode 2 and a silver-silver chloride electrode can be used for the second pseudo reference electrode 3 .
  • ion current can be generated.
  • a sample 22 such as a biological sample is contained in the electrolyte solution S, and the glass pipette 1 is brought close to the sample 22 using a three-axis stage, which will be described later, the ion current flowing through the tip of the glass pipette is blocked from a certain point when the tip 1A of the glass pipette 1 approaches the sample 22, and the ion current is reduced.
  • the surface shape of the sample 22 can be measured by controlling the movement of the tip of the glass pipette using the phenomenon that the ion current decreases due to the proximity between the tip of the glass pipette and the sample 22 as an indicator.
  • a manual 3-axis stage 27 is mounted on an inverted microscope 26 placed on an anti-vibration table 25 shown in FIG. 2A.
  • the Y-axis can be defined to intersect the X-axis at a 90° angle
  • the Z-axis can be defined as an axis intersecting the x-axis and the Y-axis at a 90° angle.
  • a Z coarse movement actuator 28 is provided supported by a manual 3-axis stage 27 .
  • the Z coarse movement actuator 28 has a maximum movable range of ten and several millimeters, and is a driving device capable of controlling movement in the Z direction in units of ⁇ m.
  • a Z piezo stage 29 is provided supported by a Z coarse motion actuator 28 .
  • the Z piezo stage 29 is a driving device that has a maximum movable range of several micrometers and can control movement in the Z direction with a resolution of 1 nm or less.
  • an XY coarse actuator 30 having a maximum movable range of several tens of millimeters and capable of controlling movement in the XY direction in units of several tens of nanometers
  • an XY piezo stage 31 having a maximum movable range of several tens of micrometers and using piezoelectric elements that can be controlled with a resolution of 1 nm or less are installed.
  • a minute current measuring instrument 19 capable of detecting and recording the ion current that can be obtained from the glass pipette 1 is installed via a first wiring 18, and has an electromagnetic shield 33 capable of shielding the minute current measuring instrument 19 including the glass pipette 1 and the first pseudo reference electrode from electromagnetic noise from the outside.
  • the XY coarse movement actuator 30 and the XY piezo stage 31 have an opening in the central part, and an accommodation part 34 is formed in which the bottom surface of the sample is the focal length of the inverted microscope.
  • the glass pipette 1 can approach or separate from the sample 22 accommodated in the container 7 along the X-axis direction, the Y-axis direction, and the Z-axis direction.
  • an objective lens 35 that functions as an inverted microscope 26 to obtain an enlarged image of a sample is installed near the housing portion 34 .
  • FIG. 2A only the container 7 and the glass pipette 1 are drawn on the XY piezo stage 31, but in the same way as the structure shown in FIG.
  • the scanning ion conductance microscope K configured as shown in FIGS. 1, 2A, and 2B uses a Z piezo stage 29, a Z coarse motion actuator 28, an XY piezo stage 31, and an XY coarse motion actuator 30, so that the tip of the glass pipette 1 can be moved to any position in the X-axis direction, the Y-axis direction, and the Z-axis direction within the electrolyte solution S contained in the container 7.
  • the XY coarse motion actuator 30 and Z coarse motion actuator 28 constitute a three-axis primary stage
  • the XY piezo stage 31 and Z piezo stage 29 constitute a three-axis secondary stage.
  • the position where the ion current has decreased by a specified amount is estimated to be very close to the surface of the sample 22, then the tip of the glass pipette 1 is slightly separated from the surface of the sample 22, the glass pipette 1 is moved slightly (several tens of nm level) in the horizontal direction of the surface of the sample 22, and the tip of the glass pipette 1 is brought closer to the surface of the sample 22 again at that position.
  • the glass pipette 1 can be moved by minute distances on the order of ⁇ m or nm by using the three-axis primary stage and the three-axis secondary stage. Moreover, the observation image of the surface of the sample 22 can be obtained by scanning the tip of the glass pipette 1 without contacting the surface of the sample 22 . Therefore, even if the sample 22 is a fine biological sample such as a cell, an observation image of the sample surface can be acquired without damaging the sample 22 .
  • FIG. 3 is a graph showing an example of a state in which the ion current is inhibited according to the distance between the tip of the glass pipette 1 and the sample 22.
  • the horizontal axis indicates the distance between the pipette and the sample
  • the normalized current value indicated on the vertical axis indicates the normalized value of the current value measured by the minute current measuring device 19 .
  • the ion current is measured while the tip of the glass pipette 1 is brought close to the surface of the sample 22. For example, as shown in FIG.
  • the surface shape of the sample 22 can be grasped.
  • the resolving power non-invasive, high spatial resolution
  • suction and discharge are performed by applying hydraulic pressure to the pipette 1, but instead of hydraulic pressure, air pressure may be used to control suction and discharge by applying air pressure. Also, instead of hydraulic pressure, other fluid may be used to control suction and discharge using means such as applying fluid pressure.
  • the graph in FIG. 4 shows the relationship between the voltage (V) applied between the first pseudo reference electrode 2 and the second pseudo reference electrode 3 and the current (nA) measured when the pressure (5.5 kPa, 7.5 kPa, 9.5 kPa) applied by the pump 6 is adjusted using a glass pipette 1 having an inlet 1a with an inner diameter of 70 nm. Since the electrical resistance of the pipette decreases according to the amount of solution collected, it is possible to roughly estimate the amount collected from the current value and voltage. With the configuration shown in FIGS. 1, 2A, and 2B, this pipette can collect tens of thousands to hundreds of fL of electrolyte solution at the tip of the glass pipette. As shown in FIG.
  • the current history during inhalation (indicated by arrow e1 ) and the current history during ejection (indicated by arrow e2 ) are slightly different, but generally exhibit linearity.
  • the increase in pressure reduces the current value and improves the linearity. Since the voltage can be controlled in sub-mV units, the recovery amount can be controlled at the aL level.
  • the amount of liquid sucked from the suction port 1a of the glass pipette 1 is maximized in the range of about 0 to 0.2 V, but when the potential is shifted from that potential to the positive side or the negative side, the liquid is discharged from the suction port 1a.
  • This phenomenon is attributed to the change in tension at the liquid-liquid interface on the tip side of the glass pipette 1 due to the action of the voltage. Therefore, the maximum recovery/discharge amount is determined by the shape of the pipette, and the recovery amount can be controlled by applying a voltage of approximately -1 V or more to 1 V or less.
  • the glass pipette 1 can be used as an electrochemical syringe. For example, as shown in FIG. 1, by inserting the glass pipette 1 into the sample 22 and controlling the voltage, part of the biological sample can be sucked into the glass pipette 1 and recovered. Further, by extracting the glass pipette 1 from the sample 22 and moving it to another place and then performing voltage control again, it is possible to eject a part of the previously sucked sample 22 from the tip of the glass pipette 1 .
  • the tip diameter R 0 when the tip diameter R 0 is small, the rate at which ⁇ shown in formula (2) increases during suction increases, and when the angle of the tip changes, the recovery amount deviates greatly from the linearity as in the research results of Anumita Saha-Shah et al.
  • the tip portion 1A tends to be short, and it is difficult to achieve both a long tip portion 1A and a small tip diameter R0 .
  • FIG. 6 shows the recovery amount of liquid that can be sucked into the glass pipette by performing the above-described voltage control while applying pressure (kPa) applied from the pump 6 when using a glass pipette having a diameter of 700 nm in the examples described later.
  • FIG. 7 shows the amount of liquid that can be sucked into the glass pipette by controlling the voltage as described above while applying pressure (kPa) applied from the pump 6 when using a glass pipette with a diameter of 100 nm.
  • FIG. 8 shows the recovery amount of the liquid that can be sucked into the glass pipette by performing the voltage control described above while applying pressure (kPa) applied from the pump 6 when using a glass pipette with a diameter of 70 nm.
  • FIGS. 6 to 8 show, in the recovery device A shown in FIG. 1, a nanopipette with an inlet diameter set to 70 to 700 nm is used, and by controlling the pressure applied from the pump 6 to the glass pipette 1 and performing the voltage control described above, the maximum volume of liquid recovered, which was determined by the shape of the pipette in the electrochemical syringe and could not be changed in the prior art, was found to be precisely controlled in the recovery device A between several tens of fL to 100,000 fL. . That is, by controlling the voltage while applying pressure to the inside of the glass pipette 1 by the pump 6, the liquid can be sucked and discharged within the ranges shown in FIGS. Since 1 nL corresponds to 1000000 (1 ⁇ 10 6 ) fL, the recovery amount of 100000 on the vertical axis in FIG. 7 corresponds to 0.1 nL.
  • An organic electrolyte solution (specific solution name: 10 mM tetrahexylammonium tetrakis(4-chlorophenyl)borate (THATPBCl), 1.2-dichloroethane solution) was filled in a pipette using the collection device A having the configuration shown in FIG.
  • THATPBCl tetrahexylammonium tetrakis(4-chlorophenyl)borate
  • 1.2-dichloroethane solution was filled in a pipette using the collection device A having the configuration shown in FIG.
  • a glass pipet with a inpettal of 700 nm in the tip + 5 kpa, + 7 kPa, + 9 kPa, + 15 kPa, + 17 kPa
  • the voltage applied between the electrode 2 and the second pseudo reference electrode 3 was sweeped within the range of -0.5 to 0 V and the amount of solution recovered by the glass pipet was measured.
  • the amount of solution recovered was obtained by observing the tip of the glass pipette from the lateral direction with an optical microscope, observing the liquid surface of the electrolyte solution aspirated with the glass pipette, approximating the shape of the tip of the glass pipette to a cone, and calculating the amount of recovery according to the position of the liquid surface by calculating the volume of the cone.
  • the above results are shown in FIG.
  • FIG. 6 by adjusting the pressure applied from the pump to the glass pipette and controlling the voltage, the maximum recovery amount could be adjusted within the range of 9.7 ⁇ 10 1 fL to 6.5 ⁇ 10 4 fL.
  • the pressure applied from the pump 6 to the glass pipette is adjusted to -15 kPa, -10 kPa, -5 kPa, 0 kPa, +2.5 kPa, +5 kPa, +7.5 kPa, +10 kPa, and the voltage applied from the voltage source 5 between the first pseudo reference electrode 2 and the second pseudo reference electrode 3 is set to -0.5 to 0 V at each pressure.
  • the recovery amount of the electrolyte solution was measured when aspirated by The above results are shown in FIG. As shown in FIG. 7, by adjusting the pressure applied from the pump to the glass pipette and controlling the voltage, it was possible to adjust the recovery amount within the range of 6.8 ⁇ 10 1 fL to 2.7 ⁇ 10 3 fL.
  • the pressure applied from the pump 6 to the glass pipette from negative pressure to positive pressure was adjusted to -7.5 kPa, -5 kPa, 0 kPa, +5 kPa, and the voltage applied from the voltage source 5 between the first pseudo reference electrode 2 and the second pseudo reference electrode 3 was set to -0.5 to 0 V at each pressure.
  • the above results are shown in FIG.
  • FIG. 8 by adjusting the pressure applied from the pump to the glass pipette and controlling the voltage, it was possible to adjust the recovery amount within the range of 5.2 ⁇ 10 2 fL to 3.7 ⁇ 10 3 fL.
  • the recovery amount can be accurately controlled and recovered over a wide range.
  • the electrolyte solution collected in the glass pipette can be discharged by adjusting the voltage applied between the first pseudo reference electrode 2 and the second pseudo reference electrode 3, the collection device A having the configuration shown in the figure can be used for discharging the solution. In this case, it is possible to increase the intake and discharge amounts compared to the conventional voltage control only, and to provide a technique capable of accurately controlling the intake and discharge amounts assuming that the intake and discharge are performed in a fine range.

Abstract

Dispositif de collecte (A) caractérisé en ce qu'il comprend une pipette (1) possédant un orifice d'aspiration (1a) à l'extrémité distale, une première électrode de pseudo-référence (2) insérée dans la pipette (1), une seconde électrode de pseudo-référence (3) immergée dans une solution électrolytique (S) où la pipette (1) est insérée, une source de tension (5) connectée à la première électrode de pseudo-référence (2) et à la deuxième électrode de pseudo-référence (3), et une pompe (6) connectée à l'extrémité proximale de la pipette (1) pour appliquer une pression dans la pipette (1).
PCT/JP2023/001520 2022-01-21 2023-01-19 Dispositif de collecte, microscope à conductance ionique à balayage qui en est équipé et procédé de collecte WO2023140324A1 (fr)

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JP2018153142A (ja) * 2017-03-17 2018-10-04 セイコーインスツル株式会社 ナノピペット
JP2019535245A (ja) * 2016-10-19 2019-12-12 ジェネラル オートメーション ラボ テクノロジーズ インコーポレイテッド ハイスループット微生物学適用高分解能システム、キット、装置、並びに微生物その他のスクリーニング方法
JP2021018147A (ja) * 2019-07-19 2021-02-15 国立大学法人金沢大学 オペランド計測を可能とした走査型イオンコンダクタンス顕微鏡

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JP2012010664A (ja) * 2010-07-02 2012-01-19 Kanagawa Acad Of Sci & Technol 細胞分析装置
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