WO2016132945A1 - Procédé de réaction et dispositif de réaction - Google Patents

Procédé de réaction et dispositif de réaction Download PDF

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Publication number
WO2016132945A1
WO2016132945A1 PCT/JP2016/053672 JP2016053672W WO2016132945A1 WO 2016132945 A1 WO2016132945 A1 WO 2016132945A1 JP 2016053672 W JP2016053672 W JP 2016053672W WO 2016132945 A1 WO2016132945 A1 WO 2016132945A1
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WIPO (PCT)
Prior art keywords
tip
pipette
pipette tip
position information
reaction
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PCT/JP2016/053672
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English (en)
Japanese (ja)
Inventor
正貴 松尾
野田　哲也
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コニカミノルタ株式会社
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Priority to JP2017500607A priority Critical patent/JPWO2016132945A1/ja
Publication of WO2016132945A1 publication Critical patent/WO2016132945A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/648—Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence

Definitions

  • the present invention uses a pipette tip attached to a pipette nozzle and a reaction chip having a recess and a seal sealing the opening of the recess to react two or more substances in the reaction chip.
  • the present invention relates to a method and a reaction apparatus.
  • SPR surface plasmon resonance
  • a detection chip having a prism made of a dielectric material, a metal film disposed on the prism, and a flow path member that is disposed on the metal film and forms a liquid flow path is used.
  • a capturing body for capturing the substance to be detected is disposed on the metal film.
  • the flow path member has an injection part for injecting a liquid such as a specimen containing a substance to be detected into the liquid flow path, and a discharge part for discharging the liquid from the liquid flow path.
  • the inlet of the inlet and the outlet of the outlet are formed in a complementary shape with respect to the tip of the pipette tip.
  • the tip of the pipette tip when the tip of the pipette tip is inserted into the inlet or outlet, the tip of the pipette tip and the inlet or outlet are fitted. Thereby, the tip of the pipette tip is arranged at a fixed position with respect to the bottom surface of the liquid channel, and the amount of liquid in the liquid channel can be controlled with high accuracy.
  • the tip of the pipette chip is arranged near the flow path in the detection chip or the bottom of the well, and the pipette chip This is done by repeatedly sucking the liquid in and discharging the liquid out of the pipette tip.
  • the pipette chip it is necessary to accurately control the positional relationship between the liquid channel or the bottom of the well and the tip of the pipette tip.
  • reaction chip when performing a plurality of reaction steps including a liquid removal step in the detection chip (reaction chip), from the viewpoint of improving the accuracy of the detection result and stabilizing the reaction efficiency, the liquid flow path in the liquid removal step or The amount of liquid remaining in the well needs to be minimized and constant. Also in this case, it is necessary to control the position of the tip of the pipette tip with high accuracy.
  • a strain gauge or a load cell as a method for detecting the position information of the tip of such a pipette tip.
  • the strain gauge is arranged between a pipette nozzle to which a pipette tip is attached and a pipette drive unit. Then, the position information of the tip of the pipette tip is acquired based on the output of the strain gauge when the pipette attached with the pipette tip is brought into contact with the reference portion that is the positioning reference of the tip of the pipette tip.
  • a pressure-sensitive sensor as another method for acquiring the position information of the tip of the pipette tip.
  • the position information of the tip of the pipette tip is acquired based on the output of the pressure sensitive sensor when the tip of the pipette tip is brought into contact with the pressure sensitive sensor.
  • a detection chip in which an inlet and an outlet are sealed with a seal may be used.
  • This detection tip may be used by piercing the seal with a pipette tip.
  • the object of the present invention is to accurately control the position of the tip of the pipette tip without increasing the manufacturing cost of the pipette tip and the reaction tip even when the seal of the reaction tip is broken by the pipette tip.
  • a reaction method uses a pipette tip attached to a pipette nozzle and a reaction tip having a recess and a seal that seals the opening of the recess.
  • a reaction method of reacting two or more substances in the reaction chip, the first step of breaking through the seal of the reaction chip with the pipette tip, and the tip of the pipette tip after the first step A second step of acquiring the position information of the second step, and after the second step, the pipette tip is operated based on the position information of the tip of the pipette tip to react two or more substances in the reaction tip. 3 steps.
  • a reaction apparatus uses a reaction chip having a recess and a seal that seals the opening of the recess, and breaks the seal with a pipette chip.
  • a reaction apparatus for reacting two or more substances in the reaction chip afterwards a chip holder for holding the reaction chip, a pipette having a pipette nozzle to which the pipette chip can be attached and detached, and a pipette for moving the pipette
  • a position information acquisition unit that acquires position information of a tip of the pipette tip.
  • the position information acquisition unit is configured such that the pipette tip moved by the pipette movement unit performs the sealing once.
  • the pipette moving unit After the information acquisition unit acquires the position information of the tip of the pipette tip, the pipette is used once or twice or more based on the position information of the tip of the pipette tip to react two or more substances in the reaction tip. Move.
  • the position of the tip of the pipette tip is accurately controlled without increasing the manufacturing cost of the pipette tip and the reaction tip.
  • Two or more substances can be appropriately reacted in the inside.
  • the presence or amount of a substance to be detected can be detected with high accuracy.
  • FIG. 1 is a schematic diagram showing the configuration of the SPFS apparatus according to the first embodiment.
  • 2A to 2C are diagrams showing the configuration of the detection chip.
  • FIG. 3 is a schematic cross-sectional view of another form of the detection chip.
  • FIG. 4 is a flowchart showing the operation of the SPFS apparatus according to the first embodiment.
  • FIG. 5 is a flowchart showing the content of the step of acquiring the position information of the tip of the pipette tip.
  • 6A is a diagram for explaining a process of breaking through a seal that closes an injection port
  • FIG. 6B is a diagram for explaining a process of breaking through a seal that closes a recess
  • FIG. It is a figure for demonstrating the process of attracting
  • FIG. 7A is a diagram illustrating a partial configuration of the SPFS apparatus according to the second embodiment
  • FIG. 7B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip
  • FIG. 8A is a diagram illustrating a partial configuration of the SPFS apparatus according to the third embodiment
  • FIG. 8B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip.
  • FIG. 9A is a diagram illustrating a configuration of a part of the SPFS device according to the fourth embodiment
  • FIG. 9B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip.
  • FIG. 9A is a diagram illustrating a configuration of a part of the SPFS device according to the fourth embodiment
  • FIG. 9B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip.
  • FIG. 10A is a diagram illustrating a partial configuration of the SPFS apparatus according to the fifth embodiment
  • FIG. 10B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip
  • FIG. 11A is a diagram illustrating a partial configuration of the SPFS device according to the sixth embodiment
  • FIG. 11B is a flowchart illustrating a process of acquiring position information of the tip of the pipette tip.
  • SPFS apparatus surface plasmon excitation enhanced fluorescence analyzer
  • FIG. 1 is a schematic diagram showing a configuration of a surface plasmon excitation enhanced fluorescence analyzer (SPFS apparatus) 100 according to Embodiment 1 of the present invention.
  • SPFS apparatus surface plasmon excitation enhanced fluorescence analyzer
  • the SPFS device (detection device) 100 includes a liquid feeding unit 110 including a pipette 111 and a pipette moving unit 112, a transport unit 120 including a tip holder 121, a position information acquisition unit 130, a light The irradiation unit 140, the light detection unit 150, and the control unit 160 are included.
  • the SPFS device 100 is used with the detection chip (reaction chip) 10 mounted on the chip holder 121. Therefore, the detection chip 10 will be described first, and then each component of the SPFS device 100 will be described.
  • FIG. 2 is a diagram illustrating a configuration of the detection chip 10.
  • 2A is a plan view of the detection chip 10
  • FIG. 2B is a cross-sectional view taken along line AA shown in FIG. 2A
  • FIG. 2C is a cross-sectional view taken along line BB shown in FIG. 2A.
  • FIG. 3 is a schematic cross-sectional view showing another form of the detection chip 10.
  • the detection chip 10 includes a prism 20 including an incident surface 21, a film formation surface 22 and an output surface 23, a metal film 30, a flow path including a reaction region 41 and a reagent storage region 42. It has a lid 40 and a seal 50.
  • the metal film 30 and the flow path lid 40 are disposed on the film formation surface 22 of the prism 20.
  • the prism 20, the metal film 30, and the channel lid 40 form a channel 60 through which liquid flows.
  • the flow path 60 is disposed directly or via the metal film 30 on the film formation surface 22 of the prism 20.
  • the detection chip 10 may be a reusable chip or a disposable chip. In the present embodiment, the detection chip 10 is a disposable chip.
  • liquid flowing through the flow path examples include a sample containing a substance to be detected (for example, blood, serum, plasma, urine, nasal fluid, saliva, semen, etc.) or a label containing a capturing substance labeled with a fluorescent substance.
  • a substance to be detected for example, blood, serum, plasma, urine, nasal fluid, saliva, semen, etc.
  • a label containing a capturing substance labeled with a fluorescent substance for example, blood, serum, plasma, urine, nasal fluid, saliva, semen, etc.
  • the prism 20 is made of an insulator that is transparent to the excitation light ⁇ . As described above, the prism 20 has the entrance surface 21, the film formation surface 22, and the exit surface 23. The incident surface 21 allows the excitation light ⁇ from the light irradiation unit 140 to enter the prism 20. A metal film 30 is disposed on the film formation surface 22. In the present embodiment, the excitation light ⁇ incident on the inside of the prism 20 is applied to the metal film 30 where the substance to be detected is captured. The excitation light ⁇ is reflected on the back surface of the metal film 30 to become reflected light ⁇ . More specifically, the excitation light ⁇ is reflected at the interface (deposition surface 22) between the prism 20 and the metal film 30 to become reflected light ⁇ . The emission surface 23 emits the reflected light ⁇ to the outside of the prism 20.
  • the shape of the prism 20 is not particularly limited.
  • the prism 20 is a pillar having a trapezoidal bottom surface.
  • the surface corresponding to one base of the trapezoid is the film formation surface 22, the surface corresponding to one leg is the incident surface 21, and the surface corresponding to the other leg is the emission surface 23.
  • the trapezoid serving as the bottom surface is preferably an isosceles trapezoid. Thereby, the entrance surface 21 and the exit surface 23 are symmetric, and the S wave component of the excitation light ⁇ is less likely to stay in the prism 20.
  • the incident surface 21 is formed so that the excitation light ⁇ does not return to the light irradiation unit 140.
  • the light source of the excitation light ⁇ is a laser diode (hereinafter also referred to as “LD”)
  • LD laser diode
  • the angle of the incident surface 21 is set so that the excitation light ⁇ does not enter the incident surface 21 perpendicularly in the scanning range centered on the enhancement angle.
  • the “enhancement angle” means scattered light having the same wavelength as the excitation light ⁇ emitted above the detection chip 10 when the incident angle of the excitation light ⁇ with respect to the metal film 30 is scanned (hereinafter referred to as “plasmon scattered light”). This means the angle of incidence when the amount of ⁇ is maximized.
  • the angle between the incident surface 21 and the film formation surface 22 and the angle between the film formation surface 22 and the emission surface 23 are both about 80 °.
  • the enhancement angle is generally determined by the design of the detection chip 10.
  • the design factors are the refractive index of the prism 20, the refractive index of the metal film 30, the film thickness of the metal film 30, the extinction coefficient of the metal film 30, the wavelength of the excitation light ⁇ , and the like.
  • the enhancement angle is shifted by the substance to be detected trapped on the metal film 30, but the amount is less than several degrees.
  • the prism 20 has a considerable amount of birefringence.
  • the material of the prism 20 include insulating resin and glass.
  • the material of the prism 20 is preferably a resin having a refractive index of 1.4 to 1.6 and a small birefringence.
  • the metal film 30 is disposed so as to be exposed to at least a part of the flow path 60 on the film formation surface 22 of the prism 20.
  • the metal film 30 causes an interaction (SPR) between the photon of the excitation light ⁇ incident on the film formation surface 22 under total reflection conditions and the free electrons in the metal film 30, and is locally on the surface of the metal film 30.
  • SPR interaction
  • In-situ light commonly referred to as “evanescent light” or “near-field light” can be generated.
  • the material of the metal film 30 is not particularly limited as long as it is a metal capable of generating SPR.
  • Examples of the material of the metal film 30 include gold, silver, copper, aluminum, and alloys thereof.
  • the metal film 30 is a gold thin film.
  • the method for forming the metal film 30 is not particularly limited. Examples of the method for forming the metal film 30 include sputtering, vapor deposition, and plating.
  • the thickness of the metal film 30 is not particularly limited, but is preferably in the range of 30 to 70 nm.
  • a capturing body for capturing a substance to be detected is fixed on the surface of the metal film 30.
  • the capturing body is uniformly fixed to a predetermined region on the metal film 30.
  • the region where the capturing body is fixed serves as a reaction field where a primary reaction and a secondary reaction described later occur.
  • the capturing body fixed to the metal film 30 is exposed in the flow path 60.
  • the type of capturing body is not particularly limited as long as it can capture the substance to be detected.
  • the capturing body is an antibody or a fragment thereof that can specifically bind to the substance to be detected.
  • the channel lid 40 is disposed on the film formation surface 22.
  • the flow path lid 40 has the reaction region 41 and the reagent storage region 42.
  • the reaction region 41 is a region for performing a primary reaction and a secondary reaction described later.
  • the reagent storage area 42 is an area in which a labeling solution used for the secondary reaction, a cleaning solution used for cleaning after each reaction, and the like are stored.
  • a channel groove 43 to be the channel 60 is formed on the back surface of the reaction region 41 in the channel lid 40.
  • a first through hole 44 serving as the injection portion 70 and a second through hole 45 serving as the storage portion 80 are opened on the front surface and the back surface of the reaction region 41, respectively.
  • Both ends of the flow channel 43 are connected to the first through hole 44 and the second through hole 45, respectively.
  • a recess 46 opened on the surface is formed in the reagent storage area 42.
  • the number of the recessed parts 46 is not specifically limited. In the present embodiment, the number of recesses 46 is two.
  • a labeling solution used for the secondary reaction, a cleaning solution, and the like are stored in the recess 46.
  • the channel groove 43, the first through hole 44, and the second through hole 45 become the channel 60, the injection unit 70, and the storage unit 80, respectively, by stacking the prism 20, the metal film 30, and the channel lid 40 in this order. .
  • the openings of the injection part 70, the storage part 80 and the four recesses 46 are respectively sealed with seals 50.
  • the channel lid 40 is preferably made of a material that is transparent to the fluorescence ⁇ emitted from the metal film 30 and the plasmon scattered light ⁇ .
  • An example of the material of the flow path lid 40 includes a resin.
  • the flow path cover 40 may be formed of an opaque material as long as the portion from which the fluorescent ⁇ and the plasmon scattered light ⁇ are extracted is transparent to the fluorescent ⁇ and the plasmon scattered light ⁇ .
  • the channel lid 40 is bonded to the prism 20 or the metal film 30 by, for example, adhesion using a double-sided tape or an adhesive, laser welding, ultrasonic welding, or pressure bonding using a clamp member.
  • the seal 50 closes all the openings of the injection part 70, the storage part 80, and the four recesses 46.
  • the material of the seal 50 can seal the opening of the injection part 70, the storage part 80, and the recess 46, respectively, and the seal 50 that seals the opening of the injection part 70 and the recess 46 can be pierced by the pipette tip 170.
  • the material of the seal 50 include an aluminum foil, an aluminum deposited film, low-density polyethylene, an elastomer material such as polyurethane, and the like.
  • the seal 50 may be a laminated seal in which a plurality of materials are laminated. In the present embodiment, the material of the seal 50 is low density polyethylene.
  • the seal 50 is configured so that a force necessary for breaking through with the pipette tip 170 is 10 N or less.
  • the force required to pierce with the pipette tip 170 varies depending on the material of the seal 50, the recess 46, the opening of the injection portion 70, the area of the opening of the storage portion 80, the shape of the tip of the pipette tip 170, and the like.
  • the force required to pierce with the pipette tip 170 can be measured using a push-pull gauge or the like.
  • the opening area of the recess 46 is 12 mm 2
  • the outer diameter of the tip of the pipette tip 170 is 1 mm (without chamfering)
  • the required force is about 7N. Note that any one of the openings of the injection unit 70, the storage unit 80, and the recess 46 may not be blocked by the seal 50.
  • the detection chip 10 ′ may have a well 60 ′ instead of the flow path 60.
  • the opening of the well 60 ′ is sealed with a seal 50.
  • liquid is injected or removed from the opening of the well 60 '.
  • the excitation light ⁇ enters the prism 20 at the incident surface 21.
  • the excitation light ⁇ that has entered the prism 20 is applied to the metal film 30 at a total reflection angle (an angle at which SPR occurs).
  • a total reflection angle an angle at which SPR occurs.
  • This localized field light excites a fluorescent substance that labels the substance to be detected present on the metal film 30 and emits fluorescence ⁇ .
  • the SPFS device 100 detects the presence or amount of the substance to be detected by measuring the amount of fluorescence ⁇ emitted from the fluorescent substance.
  • the SPFS device 100 includes the liquid feeding unit 110, the transport unit 120, the position information acquisition unit 130, the light irradiation unit 140, the light detection unit 150, and the control unit 160.
  • the detection chip 10 can be held by the chip holder 121 of the transport unit 120.
  • the liquid feeding unit 110 includes a pipette 111, a pipette moving unit 112, and a liquid feeding pump drive mechanism 113.
  • the liquid feeding unit 110 injects a specimen into the flow path 60 of the detection chip 10 held by the chip holder 121, or a liquid such as a labeling solution or a cleaning liquid stored in the reagent storage region 42 of the detection chip 10 in the reaction region. 41 in the channel 60. Further, the liquid feeding unit 110 also discharges the liquid from the flow path 60 and stirs the liquid in the flow path 60.
  • the liquid feeding unit 110 is used in a state where the pipette tip 170 is attached to the pipette nozzle 116 of the pipette 111. It should be noted that the pipette tip 170 is preferably replaceable from the viewpoint of preventing contamination of impurities.
  • the pipette 111 sucks the liquid when injecting the liquid into the flow path 60 or removing the liquid from the flow path 60.
  • the pipette 111 includes a syringe 114, a plunger 115 that can reciprocate inside the syringe 114, and a pipette nozzle 116 connected to the syringe 114. Further, the pipette 111 can quantitatively suck and discharge the liquid by the reciprocating motion of the plunger 115. Thereby, the pipette 111 can inject liquid into the flow path 60 or remove the liquid from the flow path 60. Moreover, the pipette 111 can stir the liquid in the flow path 60 by repeating suction and discharge of the liquid.
  • the pipette moving unit 112 moves the pipette nozzle 116 in order to suck the liquid into the pipette tip 170, discharge the liquid from the pipette tip 170, or break the seal 50.
  • the pipette moving unit 112 freely moves the pipette nozzle 116 in the axial direction (for example, the vertical direction) of the pipette nozzle 116.
  • the pipette moving unit 112 includes, for example, a solenoid actuator and a stepping motor.
  • the pipette moving unit 112 moves the seal 50 to be broken through the detection chip 10 to the position directly below the pipette nozzle 116 by the transport unit 120, and then seals the pipette nozzle 116 (pipette tip 170).
  • the seal 50 is broken with the pipette tip 170.
  • the distance between the seal 50 and the bottom surface of the flow path 60 see FIG. 6A; d1
  • the distance between the seal 50 and the bottom surface of the recess 46 see FIG. 6B; d3
  • the recess 46 are stored. Since the depth of the liquid (see FIG. 6B; d4) is recorded in advance, the tip of the pipette tip 170 does not come into contact with the metal film 30 in the flow path 60 or the liquid stored in the recess 46. .
  • the liquid feed pump drive mechanism 113 moves the plunger 115 to suck the external liquid into the pipette tip 170 or discharge the liquid inside the pipette 111 to the outside.
  • the liquid feed pump drive mechanism 113 includes a device for reciprocating the plunger 115 such as a stepping motor.
  • the stepping motor is preferable from the viewpoint of managing the remaining liquid amount of the detection chip 10 because it can manage the liquid feeding amount and the liquid feeding speed of the pipette 111.
  • the liquid feed pump drive mechanism 113 is preferably not driven during the process in which the pipette tip 170 breaks through the seal 50.
  • the liquid feeding unit 110 sucks various liquids from the recess 46 and injects them into the flow channel 60 of the detection chip 10.
  • the reciprocating operation of the plunger 115 with respect to the syringe 114 is repeated, so that the liquid is flown in the flow channel 60 in the detection chip 10. It reciprocates and the liquid in the flow path 60 is stirred.
  • the liquid in the channel 60 is again sucked by the pipette 111 and discharged to a waste liquid tank or the like not shown.
  • reaction with various liquids, washing, and the like can be performed, and a detection target substance labeled with a fluorescent substance can be arranged in the reaction field in the flow path 60.
  • the transport unit 120 transports the detection chip 10 to the detection position or the liquid feeding position, and holds the detection chip 10.
  • the “detection position” is a position where the light irradiation unit 140 irradiates the detection chip 10 with the excitation light ⁇ , and the light detection unit 150 detects the fluorescence ⁇ or the plasmon scattered light ⁇ generated accordingly.
  • the “liquid feeding position” is a position where the liquid feeding unit 110 injects liquid into the flow channel 60 of the detection chip 10 or removes the liquid in the flow channel 60 of the detection chip 10.
  • the transport unit 120 includes a chip holder 121 and a transport stage 122.
  • the chip holder 121 is fixed to the transfer stage 122 and holds the detection chip 10 in a detachable manner.
  • the shape of the chip holder 121 is not particularly limited as long as it can hold the detection chip 10 and does not interfere with the optical paths of the excitation light ⁇ , fluorescence ⁇ , and plasmon scattered light ⁇ .
  • the shape of the chip holder 121 is configured so that the detection chip 10 can be held with the flow path lid 40 interposed therebetween.
  • the transfer stage 122 moves the chip holder 121 in one direction and in the opposite direction (left and right direction on the paper surface of FIG. 1).
  • the transport stage 122 also has a shape that does not interfere with the optical paths of the excitation light ⁇ , fluorescence ⁇ , and plasmon scattered light ⁇ .
  • the transfer stage 122 is driven by, for example, a stepping motor.
  • the position information acquisition unit 130 acquires position information on the tip of the pipette tip 170.
  • the configuration of the position information acquisition unit 130 is not particularly limited as long as the position information of the tip of the pipette tip 170 can be acquired.
  • Examples of the position information acquisition unit 130 include a strain sensor, a pneumatic sensor 131, a capacitance sensor, a power supply and ammeter, a pressure sensor, and the like.
  • the position information acquisition unit 130 includes an air pressure sensor 131.
  • the air pressure sensor 131 is connected between the pipette nozzle 116 and the syringe 114.
  • the position information of the tip of the pipette tip 170 is obtained when gas is sucked or discharged from the tip of the pipette tip 170 while changing the distance between the tip of the pipette tip 170 and the reference portion 180 (see FIG. 6A).
  • the change in the air pressure in the pipette tip 170 is measured by the air pressure sensor 131.
  • the “reference portion” means a reference position at the tip of the pipette tip 170.
  • the reference unit 180 is not particularly limited as long as its position is specified with high accuracy, and may be a part of the detection chip 10 or a part of the SPFS device 100.
  • Examples of the reference unit 180 included in the detection chip 10 include the flow path lid 40, the seal 50, the prism 20, and the like.
  • examples of the reference unit 180 included in the SPFS apparatus 100 include a transport stage 122, a chip holder 121, and a base that supports the transport stage 122 in the transport unit 120 (portion located below the pipette nozzle 116). There may be. The suction or discharge of gas at the tip of the pipette tip 170 may be performed continuously or intermittently.
  • the light irradiation unit 140 irradiates the excitation light ⁇ toward the incident surface 21 of the detection chip 10 held by the chip holder 121. At the time of measuring the fluorescence ⁇ or the plasmon scattered light ⁇ , the light irradiation unit 140 emits only the P wave toward the incident surface 21 so that the incident angle with respect to the metal film 30 is an angle that causes SPR. .
  • the “excitation light” is light that directly or indirectly excites the fluorescent material.
  • the excitation light ⁇ is light that generates localized field light on the surface of the metal film 30 that excites the fluorescent material when the metal film 30 is irradiated through the prism 20 at an angle at which SPR occurs.
  • the light irradiation unit 140 includes a light source unit 141, an angle adjustment mechanism 142, and a light source control unit 143.
  • the light source unit 141 emits the collimated excitation light ⁇ having a constant wavelength and light amount so that the shape of the irradiation spot on the back surface of the metal film 30 is substantially circular.
  • the light source unit 141 includes, for example, a light source of excitation light ⁇ , a beam shaping optical system, an APC mechanism, and a temperature adjustment mechanism (all not shown).
  • the type of the light source is not particularly limited, and is, for example, a laser diode (LD).
  • Other examples of light sources include light emitting diodes, mercury lamps, and other laser light sources.
  • the light emitted from the light source is not a beam, the light emitted from the light source is converted into a beam by a lens, a mirror, a slit, or the like.
  • the light emitted from the light source is not monochromatic light, the light emitted from the light source is converted into monochromatic light by a diffraction grating or the like.
  • the light emitted from the light source is not linearly polarized light, the light emitted from the light source is converted into linearly polarized light by a polarizer or the like.
  • the beam shaping optical system includes, for example, a collimator, a band pass filter, a linear polarization filter, a half-wave plate, a slit, and a zoom means.
  • the beam shaping optical system may include all of these or a part thereof.
  • the collimator collimates the excitation light ⁇ emitted from the light source.
  • the band-pass filter turns the excitation light ⁇ emitted from the light source into narrowband light having only the center wavelength. This is because the excitation light ⁇ from the light source has a slight wavelength distribution width.
  • the linear polarization filter turns the excitation light ⁇ emitted from the light source into completely linearly polarized light.
  • the half-wave plate adjusts the polarization direction of the excitation light ⁇ so that the P-wave component is incident on the metal film 30.
  • the slit and zoom means adjust the beam diameter, contour shape, and the like of the excitation light ⁇ so that the shape of the irradiation spot on the back surface of the metal film 30 is a circle of a predetermined size.
  • the APC mechanism controls the light source so that the output of the light source is constant. More specifically, the APC mechanism detects the amount of light branched from the excitation light ⁇ with a photodiode (not shown) or the like.
  • the APC mechanism controls the input energy by a regression circuit, thereby controlling the output of the light source to be constant.
  • the angle adjustment mechanism 142 adjusts the incident angle of the excitation light ⁇ with respect to the metal film 30 (the interface between the prism 20 and the metal film 30 (film formation surface 22)). In order to irradiate the excitation light ⁇ at a predetermined incident angle toward a predetermined position of the metal film 30 via the prism 20, the angle adjustment mechanism 142 relatively places the optical axis of the excitation light ⁇ and the chip holder 121. Rotate.
  • the angle adjusting mechanism 142 rotates the light source unit 141 around an axis (axis perpendicular to the paper surface of FIG. 1) orthogonal to the optical axis of the excitation light ⁇ .
  • the position of the rotation axis is set so that the position of the irradiation spot on the metal film 30 hardly changes even when the incident angle is scanned.
  • the angle at which the amount of plasmon scattered light ⁇ is maximum is the enhancement angle.
  • High intensity fluorescence ⁇ can be measured by setting the incident angle of the excitation light ⁇ to the enhancement angle or an angle in the vicinity thereof.
  • the basic incident condition of the excitation light ⁇ is determined by the material and shape of the prism 20 of the detection chip 10, the film thickness of the metal film 30, the refractive index of the liquid in the flow channel 60, etc. The optimum incident condition varies slightly depending on the type and amount of the light and the shape error of the prism 20. For this reason, it is preferable to obtain an optimal enhancement angle for each measurement.
  • the light source control unit 143 controls various devices included in the light source unit 141 to control the emission of the excitation light ⁇ from the light source unit 141.
  • the light source control unit 143 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
  • the light detection unit 150 detects the amount of fluorescence ⁇ emitted from the vicinity of the surface of the metal film 30 on the flow path 60 side when the light irradiation unit 140 irradiates the metal film 30 of the detection chip 10 with the excitation light ⁇ . To do. Further, as necessary, the light detection unit 150 also detects plasmon scattered light ⁇ generated by irradiation of the excitation light ⁇ to the metal film 30.
  • the light detection unit 150 includes a light receiving unit 151, a position switching mechanism 152, and a sensor control unit 153.
  • the light receiving unit 151 is disposed in the normal direction to the surface of the metal film 30 of the detection chip 10.
  • the light receiving unit 151 includes a first lens 154, an optical filter 155, a second lens 156, and a light receiving sensor 157.
  • the first lens 154 is, for example, a condensing lens, and condenses light emitted from the metal film 30.
  • the second lens 156 is an imaging lens, for example, and forms an image of the light collected by the first lens 154 on the light receiving surface of the light receiving sensor 157.
  • the optical path between the first lens 154 and the second lens 156 is substantially parallel.
  • the optical filter 155 is disposed between the first lens 154 and the second lens 156.
  • the optical filter 155 guides only the fluorescence component to the light receiving sensor 157 and removes the excitation light component (plasmon scattered light ⁇ ) in order to detect the fluorescence ⁇ with a high S / N ratio.
  • Examples of the optical filter 155 include an excitation light reflection filter, a short wavelength cut filter, and a band pass filter.
  • the optical filter 155 is, for example, a filter including a multilayer film that reflects a predetermined light component, or a color glass filter that absorbs a predetermined light component.
  • the light receiving sensor 157 detects fluorescence ⁇ and plasmon scattered light ⁇ .
  • the light receiving sensor 157 has a high sensitivity capable of detecting weak fluorescence ⁇ from a very small amount of a substance to be detected.
  • the light receiving sensor 157 is, for example, a photomultiplier tube (PMT) or an avalanche photodiode (APD).
  • the position switching mechanism 152 switches the position of the optical filter 155 on or off the optical path in the light receiving unit 151. Specifically, when the light receiving sensor 157 detects the fluorescence ⁇ , the optical filter 155 is disposed on the optical path of the light receiving unit 151, and when the light receiving sensor 157 detects the plasmon scattered light ⁇ , the optical filter 155 is placed on the light receiving unit 151. Placed outside the optical path.
  • the sensor control unit 153 controls detection of an output value of the light receiving sensor 157, management of sensitivity of the light receiving sensor 157 based on the detected output value, change of sensitivity of the light receiving sensor 157 for obtaining an appropriate output value, and the like.
  • the sensor control unit 153 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
  • the controller 160 controls the liquid feed pump drive mechanism 113, the transport stage 122, the angle adjustment mechanism 142, the light source controller 143, the position switching mechanism 152, and the sensor controller 153.
  • the control unit 160 includes, for example, a known computer or microcomputer including an arithmetic device, a control device, a storage device, an input device, and an output device.
  • FIG. 4 is a flowchart illustrating an example of an operation procedure of the SPFS apparatus 100.
  • FIG. 5 is a flowchart showing the contents of the step (step S120 in FIG. 4) of acquiring the position information of the tip of the pipette tip 170.
  • 6A is a diagram for explaining a process of breaking through the seal 50 blocking the injection portion 70
  • FIG. 6B is a diagram for explaining a process of breaking through the seal 50 closing the recess 46
  • 6C is a diagram for explaining a step of sucking the liquid from the recess 46.
  • the primary antibody is immobilized on the metal film 30 as a capturing body.
  • a secondary antibody labeled with a fluorescent substance is used as a capturing body used for fluorescent labeling.
  • the injection part 70, the storage part 80, and the concave part 46 are each closed by a seal 50.
  • the reference portion 180 is the bottom surface of the flow path 60.
  • step S110 preparation for measurement is performed (step S110). Specifically, the detection chip 10 is prepared, and the detection chip 10 is installed in the chip holder 121 at the set position of the detection chip 10. A pipette tip 170 is attached to the tip of the pipette nozzle 116.
  • step S120 the position information of the tip of the pipette tip 170 is acquired (step S120).
  • the seal 50 having the maximum force required to pierce is pierced by the pipette tip 170 (step S121).
  • the control unit 160 operates the transport stage 122 to move the detection chip 10 to the liquid feeding position.
  • the control unit 160 drives the pipette moving unit 112 to break through the pipette tip 170 with the seal 50 having the maximum force required to pierce among the plurality of seals 50 to be pierced by the pipette tip 170.
  • the seal 50 having the maximum force necessary for piercing is the seal 50 closing the opening of the injection portion 70.
  • the distance d1 between the back surface of the seal 50 and the bottom surface of the flow path 60 is stored in advance (FIG. 6A). Therefore, the pipette moving unit 112 controls so that the tip of the pipette tip 170 does not contact the bottom surface of the flow path 60.
  • the distance between the seal 50 and the bottom surface of the recess 46 (see FIG. 6B; d3) and the depth of the liquid stored in the recess 46 (see FIG. 6B; d4) are stored in advance. Yes. Therefore, when the seal 50 having the maximum force required to pierce among the plurality of seals 50 to be pierced by the pipette tip 170 is the seal 50 closing the opening of the recess 46, the pipette moving unit 112 has the pipette tip 112. Control is performed so that the tip of 170 does not come into contact with the liquid stored in the recess 46.
  • the control unit 160 drives the pipette moving unit 112 to move the tip of the pipette tip 170 toward the bottom surface (reference unit 180) of the flow channel 60.
  • the controller 160 drives the liquid feed pump drive mechanism 113 to advance the plunger 115 relative to the syringe 114. That is, the control unit 160 brings the tip of the pipette tip 170 closer to the reference unit 180 while continuously ejecting air from the tip of the pipette tip 170.
  • the control unit 160 acquires position information of the tip of the pipette tip 170 with respect to the reference unit 180 when the air pressure sensor 131 detects the air pressure.
  • the tip of the pipette tip 170 may be brought close to the reference portion 180 while air is intermittently ejected from the tip of the pipette tip 170. Further, the tip of the pipette tip 170 may be brought close to the reference portion 180 while sucking air from the tip of the pipette tip 170 continuously or intermittently. Even in these cases, the position information of the tip of the pipette tip 170 with respect to the reference portion 180 can be obtained with high accuracy.
  • the step of breaking through the seal 50 is preferably performed before performing an operation in which the position accuracy of the pipette tip 170 is most required in all steps included in the detection operation (reaction method) of the SPFS device 100. .
  • the pipette tip 170 has the seal 50 having the maximum force required for piercing. You may make it break through.
  • the operation for which the position accuracy of the pipette tip 170 is most required is a step of removing the specimen from the flow path 60 in the primary reaction.
  • step S120 of acquiring the position information of the tip of the pipette tip is performed before the primary reaction (step S140).
  • step S120 the step of acquiring the position information of the tip of the pipette tip 170 (step S120) may be performed before the cleaning liquid is removed from the metal film 30.
  • a plurality of seals 50 may be broken.
  • the other seal 50 may be pierced, or after piercing through the other seal 50, the force required to pierce is the maximum.
  • the seal 50 may be pierced.
  • the position information of the tip of the pipette tip 170 is acquired after breaking the seal 50 once or twice or more.
  • the incident angle of the excitation light ⁇ is determined (step S130). Specifically, the control unit 160 operates the transfer stage 122 to move the detection chip 10 to the detection position. The controller 160 drives the sensor controller 153 to detect the plasmon scattered light ⁇ by the light receiving sensor 157 while driving the angle adjusting mechanism 142 to scan the incident angle of the excitation light ⁇ .
  • the angle at which the amount of plasmon scattered light ⁇ is maximized is defined as the incident angle (enhancement angle) of the excitation light ⁇ .
  • tip of the pipette tip 170 and the process (process S130) which determines the incident angle of excitation light (alpha) is not limited to this.
  • the step of determining the incident angle of the excitation light ⁇ step S130
  • the step of acquiring the position information of the tip of the pipette tip 170 step S120) may be performed.
  • step S140 the detection target substance in the specimen is reacted with the primary antibody (primary reaction; step S140).
  • the control unit 160 operates the transport stage 122 to move the container in which the specimen is stored directly below the pipette tip 170. Then, the tip of the pipette tip 170 is moved toward the container in which the specimen is stored, and the specimen is inhaled into the pipette tip 170.
  • the controller 160 operates the transfer stage 122 to move the detection chip 10 to the liquid feeding position. Then, the control unit 160 drives the pipette moving unit 112 to move the tip of the pipette tip 170 into the injection unit 70 and injects the sample into the flow channel 60.
  • the distance (see FIG. 6A; d2) between the tip of the pipette tip 170 and the bottom surface of the flow path 60 can be controlled with high accuracy. Therefore, the distance between the tip of the pipette tip 170 and the bottom surface of the flow path 60 is too large, and the specimen may adhere to the side wall of the injection unit 70. Therefore, the bottom surface (metal film 30) of the flow path 60 is not damaged.
  • a substance to be detected exists in the sample, at least a part of the substance to be detected binds to the primary antibody. After the primary reaction, the specimen is removed from the flow path 60.
  • the tip of the pipette tip 170 is brought close to the bottom surface of the channel 60 based on the position information of the tip of the pipette tip 170. Then, the sample is removed from the flow path 60 by inhaling the sample into the pipette tip 170.
  • the container in which the specimen is stored may be disposed on the detection chip 10.
  • an accommodation hole for accommodating the container is formed in the flow path lid 40 of the detection chip 10.
  • the types of the specimen and the substance to be detected are not particularly limited.
  • the specimen include body fluids such as blood, serum, plasma, urine, nasal fluid, saliva, semen, and diluted solutions thereof.
  • substances to be detected include nucleic acids (such as DNA and RNA), proteins (such as polypeptides and oligopeptides), amino acids, carbohydrates, lipids, and modified molecules thereof.
  • the specimen may be reciprocated in the flow path 60.
  • the tip of the pipette tip 170 is brought close to the bottom surface of the channel 60 based on the positional information of the tip of the pipette tip 170 as in the step of injecting the sample into the channel 60.
  • the plunger 115 is reciprocated while the position of the tip of the pipette tip 170 is fixed. Accordingly, the sample can be reciprocated in the flow path 60 by repeatedly inhaling and discharging the sample with the pipette tip 170. After reciprocating the sample in the flow channel 60, the sample is removed from the flow channel 60 by inhaling the sample into the pipette tip 170.
  • the metal film 30 is cleaned with a cleaning solution such as a buffer solution.
  • the controller 160 moves the tip of the pipette tip 170 toward the cleaning liquid in the recess 46 and causes the cleaning liquid to be sucked into the pipette tip 170.
  • the distance between the tip of the pipette tip 170 and the surface of the cleaning liquid (see FIG. 6C; d5) and the tip of the pipette tip 170 and the recess 46 The distance from the bottom surface (see FIG. 6C; d6) can also be controlled with high accuracy. Therefore, the cleaning liquid can be appropriately sucked into the pipette tip 170.
  • the control unit 160 drives the pipette moving unit 112 to move the tip of the pipette tip 170 into the injection unit 70 and injects the cleaning liquid into the flow channel 60.
  • the washing liquid containing the substance that has not bound to the primary antibody is removed from the flow path 60.
  • the control unit 160 drives the pipette moving unit 112 to move the tip of the pipette tip 170 to the injection unit 70.
  • the tip of the pipette tip 170 is brought close to the bottom surface of the channel 60 to remove the cleaning liquid from the channel 60.
  • the tip of the pipette tip 170 is brought close to the prism 20 (metal film 30) based on the position information of the tip of the pipette tip 170 described above, the amount of liquid remaining in the flow path 60 is minimized. be able to.
  • the position of the tip of the pipette tip 170 when removing the cleaning liquid is preferably the same as the position of the tip of the pipette tip 170 in the step of removing the specimen from the flow path 60. Thereby, the amount of liquid remaining in the flow path 60 can be made constant.
  • the target substance captured on the metal film 30 is labeled with a fluorescent substance (secondary reaction; step S150).
  • the control unit 160 moves the tip of the pipette tip 170 toward the concave portion 46 in which a liquid (labeling liquid) containing a capturing body labeled with a fluorescent substance is stored, and labels the pipette tip 170 in the pipette tip 170. Inhale fluid.
  • the position information of the pipette tip 170 is specified with high accuracy, the distance between the tip of the pipette tip 170 and the surface of the labeling liquid (see d5 in FIG.
  • the control unit 160 drives the pipette moving unit 112 to move the tip of the pipette tip 170 into the injection unit 70 and injects the labeling solution into the flow channel 60.
  • the detection target substance captured on the metal film 30 is labeled with a fluorescent substance by an antigen-antibody reaction. Thereafter, the labeling liquid in the flow path 60 is removed, and the inside of the flow path 60 is cleaned with a cleaning liquid.
  • the position of the tip of the pipette tip 170 when removing the labeling liquid in the flow channel 60 is determined based on the position information of the tip of the pipette tip 170 described above. As a result, the amount of liquid remaining in the flow path 60 can be kept to a minimum and constant.
  • the order of the primary reaction (step S140) and the secondary reaction (step S150) is not limited to this.
  • the liquid containing these complexes is deposited on the metal film 30. May be provided. Further, the specimen and the labeling solution may be provided on the metal film 30 at the same time.
  • the pipette 111 is moved once or twice or more based on the position information of the tip of the pipette chip 170.
  • a substance to be detected is detected (step S160). Specifically, the control unit 160 operates the transfer stage 122 to move the detection chip 10 to the detection position.
  • the sensor control unit 153 is driven to drive the metal film 30 (while the light source control unit 143 is driven to irradiate the excitation light ⁇ to a predetermined position of the metal film 30 at the incident angle (enhancement angle) determined in step S130.
  • the light receiving sensor 157 is controlled so as to detect the intensity of the fluorescence ⁇ emitted from the surface of the metal film 30 and the vicinity thereof.
  • control part 160 may measure a blank value before a secondary reaction (process S150).
  • the excitation light ⁇ is irradiated onto the metal film 30 at an enhancement angle, and the detection value of the light receiving sensor 157 is set as a blank value.
  • the amount of fluorescence ⁇ indicating the amount of the substance to be detected in the sample is calculated by subtracting the blank value from the detected value of fluorescence ⁇ .
  • the SPFS device 100 acquires the position information of the tip of the pipette tip 170 after breaking through the seal 50 with the pipette tip 170, the position information of the tip of the pipette tip 170 is obtained.
  • the position of the tip end of the pipette tip 170 can be controlled with high accuracy without the position of the tip end of the pipette tip 170 being shifted after obtaining the above.
  • the precision of a detection result can be improved by making constant the residual liquid amount in the flow path 60.
  • the SPFS device according to the second embodiment differs from the SPFS device 100 according to the first embodiment in that the configuration of the position information acquisition unit 230 is different. Therefore, the same components as those of the SPFS device 100 according to the first embodiment are denoted by the same reference numerals, description thereof is omitted, and different portions are mainly described.
  • FIG. 7A is a diagram illustrating a partial configuration of the SPFS apparatus according to the second embodiment.
  • the position information acquisition unit 230 includes a strain sensor 231.
  • the strain sensor 231 is disposed on the pipette nozzle 116.
  • the position information of the tip of the pipette tip 170 is acquired by measuring the strain of the pipette nozzle 116 when the tip of the pipette tip 170 and the reference unit 180 come into contact with each other by the strain sensor 231.
  • the pipette tip 170 and the pipette nozzle 116 to which the pipette tip 170 is attached are distorted.
  • the contact between the tip of the pipette tip 170 and the reference portion 180 can be detected. Therefore, the position information of the tip of the pipette tip 170 can be acquired.
  • FIG. 7B is a flowchart showing a process of acquiring the position information of the tip of the pipette tip 170.
  • the force required to break through the plurality of seals 50 that the pipette tip 170 should break through is maximized.
  • the seal 50 is broken with the pipette tip 170 (step S121).
  • the distortion of the pipette nozzle 116 is measured (step S222).
  • the control unit 160 operates the pipette moving unit 112 to move the tip of the pipette tip 170 toward the bottom surface (reference unit 180) of the channel 60.
  • the pipette tip 170 and the pipette nozzle 116 to which the pipette tip 170 is attached are distorted.
  • the position information of the tip of the pipette tip 170 is acquired by detecting the contact between the tip of the pipette tip 170 and the reference portion 180 by the strain sensor 231 disposed in the pipette nozzle 116.
  • the SPFS device according to the second embodiment has the same effects as the SPFS device 100 according to the first embodiment.
  • the SPFS apparatus 100 according to the third embodiment is different from the SPFS apparatus 100 according to the first embodiment in that the configuration of the position information acquisition unit 330 is different. Therefore, the same components as those of the SPFS device 100 according to Embodiment 1 are denoted by the same reference numerals, and the description thereof is omitted.
  • FIG. 8A is a diagram illustrating a partial configuration of the SPFS apparatus according to the third embodiment.
  • the position information acquisition unit 330 has a capacitance sensor 331.
  • the position where the capacitance sensor 331 is disposed is not particularly limited. In the present embodiment, the capacitance sensor 331 is disposed on the transfer stage 122. Acquisition of the position information of the tip of the pipette tip 170 is performed between the tip of the pipette tip 170 and the reference portion 180 when the tip of the pipette tip 170 is brought close to the reference portion 180 (detection surface of the capacitance sensor 331). The change in the electrostatic capacity is measured by the electrostatic capacity sensor 331.
  • FIG. 8B is a flowchart showing a process of acquiring position information of the tip of the pipette tip 170.
  • the force required to pierce out of the plurality of seals 50 to be pierced by the pipette tip 170 is maximized.
  • the seal 50 is broken with the pipette tip 170 (step S121).
  • step S322 the change in capacitance between the tip of the pipette tip 170 and the reference portion is measured (step S322).
  • the control unit 160 operates the pipette moving unit 112 to move the tip of the pipette tip 170 toward the detection surface (reference unit 180) of the capacitance sensor 331.
  • the capacitance between the tip of the pipette tip 170 and the reference portion 180 changes.
  • position information of the tip of the pipette tip 170 is acquired.
  • the SPFS device according to the third embodiment has the same effects as the SPFS device 100 according to the first embodiment.
  • the SPFS apparatus according to the fourth embodiment differs from the SPFS apparatus 100 according to the first embodiment in that the configuration of the position information acquisition unit 430 is different. Therefore, the same components as those of the SPFS device 100 according to the first embodiment are denoted by the same reference numerals, description thereof is omitted, and different portions are mainly described.
  • FIG. 9A is a diagram illustrating a partial configuration of the SPFS apparatus according to the fourth embodiment.
  • the position information acquisition unit 430 includes a power source 431 and an ammeter 432.
  • the power source 431 and the ammeter 432 are connected to the tip of the pipette tip 170 and the reference unit 180, respectively.
  • the pipette tip 170 uses a conductive pipette tip containing carbon or the like.
  • the position information of the tip of the pipette tip 170 is acquired by applying a voltage between the tip of the pipette tip 170 and the reference portion 180 (metal film 30) by the power source 431, and using the ammeter 432 for the tip of the pipette tip 170 and the reference. This is done by measuring the impedance characteristic between the portion 180 (metal film 30).
  • FIG. 9B is a flowchart showing a process of acquiring position information of the tip of the pipette tip 170.
  • the force required to break through the plurality of seals 50 that the pipette tip 170 should break through is maximized.
  • the seal 50 is broken with the pipette tip 170 (step S121).
  • the control unit 160 operates the pipette moving unit 112 to move the tip of the pipette tip 170 toward the bottom surface (reference unit 180) of the channel 60.
  • a voltage is applied between the tip of the pipette tip 170 and the reference portion 180 (metal film 30) by the power source 431, and the pipette tip 170 is drawn by the ammeter 432.
  • the position information of the tip of the pipette tip 170 is obtained by measuring the impedance characteristic between the tip of the pipe and the reference portion 180 (metal film 30).
  • the SPFS device according to the fourth embodiment has the same effects as the SPFS device 100 according to the first embodiment.
  • the SPFS device according to the fifth embodiment is different from the SPFS device 100 according to the first embodiment in that the configuration of the position information acquisition unit 530 is different. Therefore, the same components as those of the SPFS device 100 according to the first embodiment are denoted by the same reference numerals, description thereof is omitted, and different portions are mainly described.
  • FIG. 10A is a diagram illustrating a partial configuration of the SPFS apparatus according to the fifth embodiment.
  • the position information acquisition unit 530 includes a pressure-sensitive sensor 531.
  • the position where the pressure-sensitive sensor 531 is disposed is not particularly limited. In the present embodiment, the pressure-sensitive sensor 531 is disposed on the transfer stage 122.
  • the position information of the tip of the pipette tip 170 is acquired by measuring the pressure when the tip of the pipette tip 170 comes into contact with the reference portion 180 (detection surface of the pressure sensor 531) by the pressure sensor 531.
  • the pressure-sensitive sensor 531 detects a predetermined pressure. Thereby, the contact between the tip of the pipette tip 170 and the reference portion 180 can be detected.
  • FIG. 10B is a flowchart showing a process of acquiring the position information of the tip of the pipette tip 170.
  • the force required to break through the plurality of seals 50 that the pipette tip 170 should break through is maximized.
  • the seal 50 is broken with the pipette tip 170 (step S121).
  • step S522 the contact pressure between the tip of the pipette tip 170 and the reference portion 180 is measured (step S522).
  • the control unit 160 operates the pipette moving unit 112 to move the tip of the pipette tip 170 toward the bottom surface (reference unit 180) of the channel 60.
  • the pressure-sensitive sensor 531 detects the pressure at that time. At this time, the pressure sensor 531 detects a predetermined pressure to acquire position information of the tip of the pipette tip 170.
  • the SPFS device according to the fifth embodiment has the same effect as the SPFS device 100 according to the first embodiment.
  • the SPFS device according to the sixth embodiment differs from the SPFS device 100 according to the first embodiment in that the configuration of the position information acquisition unit 630 is different. Therefore, the same components as those of the SPFS device 100 according to the first embodiment are denoted by the same reference numerals, description thereof is omitted, and different portions are mainly described.
  • FIG. 11A is a diagram illustrating a partial configuration of the SPFS apparatus according to the sixth embodiment.
  • the position information acquisition unit 630 includes an optical sensor 631.
  • the optical sensor 631 is not particularly limited as long as it can optically acquire the position information of the tip of the pipette tip 170.
  • An example of the optical sensor 631 may be a method of detecting by the presence / absence of laser light passing between the light emitting element and the light receiving element, or a method of detecting by imaging with a CCD camera.
  • the optical sensor 631 is a method of detecting by imaging with a CCD camera.
  • the position where the optical sensor 631 is arranged is not particularly limited. In the present embodiment, the optical sensor 631 is fixed to the transfer stage 122 via a support member.
  • Acquisition of the position information of the tip of the pipette tip 170 is performed by imaging when the tip of the pipette tip 170 enters the imaging region of the optical sensor 631.
  • the optical sensor 631 detects and images the tip of the pipette tip 170. Thereby, the position information of the tip of the pipette tip 170 can be acquired.
  • FIG. 11B is a flowchart showing a process of acquiring the position information of the tip of the pipette tip 170.
  • the force required for breaking through the plurality of seals 50 that the pipette tip 170 should break through is maximized.
  • the seal 50 is broken with the pipette tip 170 (step S121).
  • the tip position of the pipette tip 170 is detected (step S622).
  • the control unit 160 operates the pipette moving unit 112 to move the tip of the pipette tip 170 toward the imaging area of the optical sensor 631.
  • the optical sensor 631 detects that the tip of the pipette tip 170 has entered the imaging area of the optical sensor 631, captures the tip of the pipette tip 170, and acquires positional information of the tip of the pipette tip 170.
  • the SPFS device according to the sixth embodiment has the same effect as the SPFS device 100 according to the first embodiment.
  • the position information of the tip of the pipette tip 170 may be acquired in a state where the labeling solution or the cleaning solution is sucked into the pipette tip 170.
  • the acquisition of the position information of the tip of the pipette tip 170 and the injection operation of the labeling liquid and the cleaning liquid into the flow channel 60 can be performed simultaneously, and the detection time can be shortened. Further, in Embodiments 1 to 4 and 6, the SPFS apparatus is not contaminated.
  • the detection apparatus including the reaction apparatus according to the present invention has been described.
  • a configuration for detecting a substance to be detected for example, the conveyance unit 120, the light irradiation unit 140, and the light detection unit 150).
  • Etc. may not be included.
  • the detection method may not include the detection step (step S160).
  • the present invention is applied to the SPFS device 100 using SPR as a detection device including a reaction device.
  • the present invention is applied to an SPR device (detecting reflected light ⁇ ) using SPR. May be.
  • the same effect as in the first to sixth embodiments can be obtained.
  • the detection apparatus performs measurement by the total reflection measurement (ATR) method or the like without using SPR, but the same effects as in the first to sixth embodiments can be obtained.
  • ATR total reflection measurement
  • the reaction method and reaction apparatus according to the present invention can measure a substance to be detected with high reliability. Therefore, it is expected to contribute to the development, spread and development of a very simple quantitative immunoassay system.

Abstract

La présente invention concerne un procédé de réaction dans lequel un embout de pipette fixé à une buse de pipette et un embout de réaction ayant un évidement et un scellement scellant l'ouverture de l'évidement sont utilisés pour amener deux substances ou plus à réagir à l'intérieur de l'embout de réaction. Dans un premier temps, le scellement de l'embout de réaction est percé par l'embout de pipette (étape 1). Ensuite, après l'étape 1, des informations de position pour l'extrémité avant de l'embout de pipette sont acquises (étape 2). Après l'étape 2, l'embout de pipette est manipulé sur la base des informations de position pour l'extrémité avant de l'embout de pipette, et deux substances ou plus sont amenées à réagir à l'intérieur de l'embout de réaction (étape 3).
PCT/JP2016/053672 2015-02-20 2016-02-08 Procédé de réaction et dispositif de réaction WO2016132945A1 (fr)

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WO2019123746A1 (fr) * 2017-12-21 2019-06-27 コニカミノルタ株式会社 Système de régulation de température
JPWO2018150943A1 (ja) * 2017-02-15 2019-12-12 コニカミノルタ株式会社 送液システム、検査システム及び送液方法
WO2022196272A1 (fr) * 2021-03-16 2022-09-22 株式会社日立ハイテク Dispositif d'analyse automatique

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