WO2016143377A1 - Micropuce, puits de micropuce, dispositif d'analyse utilisant une micropuce, et procédé d'analyse utilisant une micropuce - Google Patents

Micropuce, puits de micropuce, dispositif d'analyse utilisant une micropuce, et procédé d'analyse utilisant une micropuce Download PDF

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Publication number
WO2016143377A1
WO2016143377A1 PCT/JP2016/050912 JP2016050912W WO2016143377A1 WO 2016143377 A1 WO2016143377 A1 WO 2016143377A1 JP 2016050912 W JP2016050912 W JP 2016050912W WO 2016143377 A1 WO2016143377 A1 WO 2016143377A1
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WIPO (PCT)
Prior art keywords
well
microchip
liquid
passage
inflow passage
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PCT/JP2016/050912
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English (en)
Japanese (ja)
Inventor
加藤　義明
亮輔 南
渡辺　俊夫
増原　慎
淳志 梶原
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ソニー株式会社
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Publication of WO2016143377A1 publication Critical patent/WO2016143377A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • 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
    • G01N35/08—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 stream of discrete samples flowing along a tube system, e.g. flow injection analysis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N37/00—Details not covered by any other group of this subclass

Definitions

  • the present invention relates to a microchip into which a liquid flows, a microchip well, an analysis apparatus using the microchip, and an analysis method using the microchip.
  • microchips used in the biotechnology field have wells, and reagent mixing, weighing, light detection such as fluorescence, culture, and the like are performed in the wells.
  • the well consists of a minute space, and voids are likely to occur when a liquid such as a reagent, sample, or culture solution is flowed into the well.
  • an error occurs in the mixing ratio in mixing of reagents and samples as well as in weighing.
  • the gap between the reagent and the reagent has a large difference in refractive index (approximately 0.3 to 0.4), which may cause a lens effect and reduce the light collection efficiency due to confusion, refraction, and the like.
  • refractive index approximately 0.3 to 0.4
  • the well inlet and outlet are often arranged in a straight line.
  • a liquid such as a reagent is injected from a narrow inlet into a well in a wide space
  • the injection speed is high, there is a phenomenon that the liquid jumps out as a droplet.
  • the droplet hits the outflow channel as it is, a part of the droplet passes through the outlet and a part remains in the well.
  • liquid does not accumulate in the center of the well, and there is a gap.
  • the microchip reaction apparatus includes a microchip having an internal space separated from another space by a gas-permeable wall that does not transmit liquid but allows gas to pass through, and pressure in the internal space filled with a predetermined solution. Bubble removing means for moving bubbles in the internal space to the another space by making the pressure higher than that in the other space.
  • an internal space provided in the microchip is separated from another space by a gas-permeable wall that does not transmit liquid but transmits gas. Since it is equipped with a bubble removal means that moves the bubbles in the internal space to another space by raising the pressure in the other space, even if bubbles exist in the micro space filled with the solution, It is said that the bubbles can be removed from the minute space.
  • This technology mainly aims to provide a microchip that suppresses the generation of voids in the well when the liquid flows by devising the well structure.
  • the present technology is a microchip into which a liquid flows, and has one or more wells, and an inflow passage and an outflow passage are disposed on a side surface of the well,
  • the inflow passage is disposed at an angle of 0 to 45 degrees with respect to the tangential direction or side direction of the outer edge of the well, and the liquid outlet at the upstream end of the outflow passage is the liquid at the downstream end of the inflow passage
  • a microchip is provided which is disposed at a position of an angle of 240 to 315 degrees in a liquid flow direction from a line segment connecting an inlet and a center of a well.
  • the well may have a closed curve or a closed polygon outline.
  • the inflow passage may be formed such that the opening area of the opening portion of the inflow passage widens toward the downstream side.
  • the inflow passage can be installed so that the lower side of the liquid inlet at the downstream end of the inflow passage and the bottom surface of the well are continuous.
  • the outflow passage may be disposed on the tangential side or the side direction side opposite to the flow direction of the liquid in the well.
  • the well has a top surface, and the outflow passage can be installed such that the upper side of the liquid outlet at the upstream end of the outflow passage and the top surface of the well are continuous.
  • the center of the well may be the center of gravity of the closed curve or the outer shape of the closed polygon.
  • the well may be circular.
  • the present technology is a well provided in a microchip into which a liquid is introduced, and an inflow passage and an outflow passage are disposed on a side surface of the well, and the inflow passage is in a tangential direction of an outer edge of the well
  • the liquid outlet at the upstream end of the outflow passage is arranged at an angle of 0 ° to 45 ° with respect to the side direction
  • the line connecting the liquid inlet at the downstream end of the inflow passage and the center of the well Wells can be provided that are positioned at an angle of 240 to 315 degrees from the minute to the liquid flow direction.
  • the present technology is an apparatus for analyzing a liquid sample using a microchip
  • the microchip has one or more wells, and an inflow passage and an outflow are formed on a side surface of the well.
  • the inflow passage is disposed at an angle of 0 ° to 45 ° with respect to the tangential direction or the side direction of the outer edge of the well, and the liquid outlet at the upstream end of the outflow passage It is possible to provide an analyzer that is disposed at an angle of 240 degrees to 315 degrees in the sample flow direction from a line segment connecting the liquid inlet at the downstream end of the passage and the center of the well.
  • the present technology is a method for analyzing a liquid sample using a microchip
  • the microchip has one or more wells, and an inflow passage and an outflow are formed on a side surface of the well.
  • the inflow passage is disposed at an angle of 0 ° to 45 ° with respect to the tangential direction or the side direction of the outer edge of the well, and the liquid inlet at the upstream end of the outflow passage is the inflow
  • An analysis method can be provided in which a line segment connecting the liquid inlet at the downstream end of the passage and the center of the well is disposed at an angle of 240 ° to 315 ° in the sample flow direction.
  • the liquid when a liquid is allowed to flow into a well, the liquid can be filled without creating a void.
  • the effects described here are not necessarily limited, and may be any of the effects described in the present disclosure.
  • Microchip well structure 2. First embodiment of microchip well 2. Second embodiment of microchip wells 4. Microchip manufacturing method 5. Analysis device and analysis method using microchip Example of microchip
  • the microchip of the present technology has one or more wells, an inflow passage through which liquid flows into the well, and an outflow passage through which liquid flows out of the well.
  • the well is filled with a liquid, and in the well, for example, reagent mixing, weighing, light detection such as fluorescence, culture, and the like are performed.
  • the liquid may be liquid. Examples of the liquid include distilled water, cell culture solution, physiological saline, a liquid containing primers and antibodies, a liquid in which various reagents used for chemical reaction are dissolved, or a liquid such as blood, serum, plasma, bone marrow fluid, urine, and the like.
  • the liquid includes a liquid having viscosity such as a gel that can flow into the microchip.
  • the well of the microchip of the present technology has a closed curve or closed polygonal outline.
  • the outer shape of the closed curve include a circle, an ellipse, an amoeba shape, and an outer shape of a closed curve partially including a straight line.
  • Examples of the outer shape of the closed polygon include an n-gonal shape and an n-square shape whose n is 3 or more.
  • a pentagon, a regular pentagon, a hexagon, a regular hexagon, a heptagon, a regular heptagon, an octagon, a regular octagon, and the like can be given.
  • the well may be shaped according to the plate shape.
  • the shape of the well is a cylinder having a height corresponding to the thickness of the plate, a truncated cone, an inverted truncated cone, an elliptical cylinder, an elliptical cone, an inverted elliptical truncated cone, a tapered shape, an inverted tapered shape , An n prism, an n truncated pyramid, an inverted n truncated pyramid, an n rectangular barrel taper, an inverted n rectangular barrel taper, and the like.
  • the well has a bottom surface and can also have a top surface.
  • the bottom surface and / or top surface may have grooves, steps, slopes, and the like.
  • FIG. 1 shows an example of a well structure.
  • the columnar well 1 has a top surface and a bottom surface.
  • the side of the well is provided with an inflow passage 21 and an outflow passage 31 through which liquid flows.
  • the surface where the downstream end portion of the inflow passage contacts the side surface of the well is the liquid inflow port 22, and the surface where the upstream end portion of the outflow passage contacts the side surface of the well is the liquid outflow port 32.
  • the inflow passage is arranged at an angle of 0 to 45 degrees with respect to the tangential direction or the side direction of the outer edge of the well shape when the microchip is viewed from directly above.
  • the liquid inlet or the center of the liquid inlet is the contact
  • the tangent extending from the contact is 0 degrees
  • the angle is 45 degrees in the direction away from the outer edge of the well with the contact as the center.
  • the inflow passage is arranged to extend from the contact (liquid inlet).
  • the edge extending from the liquid inlet or passing through the liquid inlet is 0 degree, and the inflow is at an angle of up to 45 degrees about the liquid inlet and away from the outer edge of the well.
  • a passage is arranged.
  • the position where the upstream end of the outflow passage is in contact with the side of the well that is, the position of the liquid outlet, connects the center of the liquid inlet or the liquid inlet and the center of the well when the microchip is viewed from directly above. From the line segment, it is disposed on the side surface of the well at a position of 240 to 315 degrees in the flow direction of the liquid entering from the liquid inlet.
  • the center of the well can be the center of gravity of the outer shape of the well. For example, if the well is circular, it becomes the center of the circle. From the liquid inlet, the liquid flows more than 240 degrees along the side of the well and reaches the liquid outlet.
  • the liquid may reach the liquid outlet after making one or more rounds in the well along the side surface of the well. At this time, the liquid may flow along the side of the well while flowing along the side of the well so as to swirl upward in the well while being pushed by the liquid flowing in later.
  • the inflow passage can be formed such that the opening area of the opening portion of the inflow passage extends toward the downstream side, that is, the liquid inlet.
  • the opening area refers to an area of a cross section when the microchip is viewed from directly above and the inflow passage is cut vertically from above.
  • the inflow passage by narrowing the opening area on the upstream side and continuously widening the opening area toward the downstream side, it is possible to prevent droplets from forming or splashing when the liquid flows into the well. it can.
  • the inflow passage can be installed so that the lower side of the liquid inlet at the downstream end of the inflow passage and the bottom surface of the well are continuous.
  • the lower side of the liquid inflow port is continuous with the bottom surface of the well so that there is no step, it is possible to prevent the liquid from being formed or scattered when flowing into the well.
  • the upper side of the liquid inlet may be continuous with the well top surface, lower than the well top surface, and may be stepped without being continuous, and is not particularly limited.
  • the outflow passage is preferably arranged in a direction opposite to the liquid flow direction.
  • a component that actively introduces liquid by suction or the like can be added to the downstream side of the outflow passage.
  • a functional film that passes only gas without passing liquid is disposed.
  • the gas can be aspirated from the outflow passage through the functional membrane to direct the liquid into the well.
  • the liquid from the inflow passage may become droplets at the liquid inflow port of the well and jump out to the outflow passage to wet the functional film.
  • the functional film gets wet, liquid filling does not proceed any more and there is almost no liquid in the well. Therefore, if the well according to the present technology described above is applied, the liquid does not become droplets or jump out, so that the functional film is not wetted, and the liquid is quickly filled without creating a void in the well. Can do.
  • FIG. 1 A first embodiment of a microchip well according to the present technology is shown in FIG.
  • the liquid flows in from the inflow passage 21 and flows around the side of the well 1 to the outflow passage 31.
  • the shape of the well 1 is a cylindrical shape
  • the arrangement of the inflow passage 21 and the outflow passage 31 is in the range of 240 ° to 315 ° in the liquid flow direction from the line segment connecting the center of the well 1 and the liquid inlet 22. It is in.
  • the angles formed between the respective tangents of the inflow passage 21 and the outflow passage 31 and the respective flow paths are in the range of 0 degree to 45 degrees.
  • the outflow passage 31 may extend in a direction opposite to the directions 51 and 52 of the liquid flowing in from the inflow passage 21 and circulating around the well 1.
  • FIG. 3 shows a second embodiment of a microchip well according to the present technology.
  • the lower side of the liquid inlet 22 at the downstream end of the inflow passage 21 is in contact with the bottom surface of the well 1, and the liquid outlet 32 at the upstream end of the outflow passage is , Having a shape in contact with the top surface of the well.
  • the bottom surface 23 of the inflow passage 21 is inclined so as to slide down the inflow passage 21 when liquid enters the well 1 from the inflow passage 21 and so that the opening area of the inflow passage 21 gradually increases.
  • the inclination angle of the bottom surface 23 of the inflow passage 21 can be set in the range of 20 degrees to 45 degrees, for example.
  • the material of the microchip can be glass or various plastics (PP, PC, COP, PDMS), and is not particularly limited.
  • PP polypropylene
  • PC polypropylene
  • COP polymethyl methacrylate
  • PDMS polymethyl methacrylate
  • the analysis using a microchip is performed optically, a material having optical transparency, low self-fluorescence, and low wavelength dispersion is selected.
  • a hard coat layer can be attached to the microchip.
  • the hard coat layer can be formed using a commonly used hard coat agent, for example, using a UV curable hard coat agent to which a fingerprint adhesion preventing agent such as a fluorine-based or silicon-based antifouling additive is added.
  • Film can be formed.
  • JP 2003-157579 A discloses a polyfunctional compound (A) having two or more polymerizable functional groups capable of being polymerized by active energy rays, an organic group having a mercapto group, and a hydrolyzable group as a hard coat agent.
  • active energy ray-curing property comprising a modified colloidal silica (B) having an average particle diameter of 1 to 200 nm, which is surface-modified with a mercaptosilane compound in which a hydroxyl group is bonded to a silicon atom, and a photopolymerization initiator (C).
  • Composition (P) is disclosed.
  • the well 1, the inflow passage 21, the outflow passage 31 and the like arranged on the microchip can be formed by wet etching or dry etching of a glass substrate layer, or by nanoimprinting, injection molding, or cutting of a plastic substrate layer. it can.
  • a microchip can be formed by covering and sealing the substrate layer formed with the well 1, the inflow passage 21, the outflow passage 31 and the like with a substrate layer of the same material or a different material.
  • the microchip according to the present technology can be applied to various analyzers using the microchip.
  • the analysis apparatus can analyze the characteristics of microparticles, etc., chemically, optically, electrically, or magnetically in a sample or the like filled in a well.
  • a liquid sample or the like is filled in the well of the microchip without having a gap and becomes an object of analysis.
  • Analyzes include, for example, microparticles.
  • animal cells such as animal cells, plant cells, microorganisms, ribosomes, liposomes, mitochondria, organelles, nucleic acids, proteins and other biologically related fine particles, latex particles and gel particles, industrial use Examples include samples containing synthetic particles such as particles, samples containing organic polymer materials, inorganic polymer materials, metals, and the like.
  • the analysis apparatus is an apparatus that performs optical analysis of a sample, for example, a light source that emits light having a wavelength selected with respect to the sample filled in the well of the microchip, and particles included in the sample And a photodetector for detecting the light separated by the spectroscopic optical system.
  • the wavelength-selected light is emitted toward the sample filled in the well of the microchip, and the well is irradiated with the light.
  • the light emitted from the microparticles contained in the sample in the well can be wavelength-separated, and the light separated by the photodetector can be detected.
  • FIG. 4 shows a photograph substituted for a drawing centering on a well portion in an example of a microchip according to the present technology.
  • the diameter of the well was 3.2 mm
  • the depth was 1.4 mm
  • the width of the main part of the inflow passage and the outflow passage was 0.4 mm
  • the depth was 0.15 mm.
  • the functional membrane 4 was installed downstream of the outflow passage 31.
  • the inflow passage 21 was formed to have a bottom surface 23, and the bottom surface of the well 1 and the lower side of the liquid inlet 22 were made continuous.
  • the functional film 4 allowed only the gas to pass therethrough, so that the liquid was sucked and the liquid flowed into the well 1. At this time, no phenomenon such as formation of droplets or scattering that caused voids was observed.
  • the liquid in the well 1 flowed along the side surface of the well 1, the liquid reached the height of the liquid outlet 32, and the liquid was filled without forming a void in the well 1.
  • a microchip into which liquid is introduced Have one or more wells, An inflow passage and an outflow passage are disposed on the side surface of the well, The inflow passage is disposed at an angle of 0 degree to 45 degrees with respect to a tangential direction or a side direction of the outer edge of the well, The liquid outlet at the upstream end of the outflow passage is disposed at an angle of 240 ° to 315 ° in the liquid flow direction from the line connecting the liquid inlet at the downstream end of the inflow passage and the center of the well.
  • To be Microchip. [2] The microchip according to [1], wherein the well has a closed curve or a closed polygonal outer shape.
  • the well has a top surface
  • a well provided in a microchip into which liquid is introduced An inflow passage and an outflow passage are disposed on the side surface of the well,
  • the inflow passage is disposed at an angle of 0 degree to 45 degrees with respect to a tangential direction or a side direction of the outer edge of the well,
  • the liquid outlet at the upstream end of the outflow passage is disposed at an angle of 240 ° to 315 ° in the liquid flow direction from the line connecting the liquid inlet at the downstream end of the inflow passage and the center of the well.
  • An apparatus for analyzing a liquid sample using a microchip has one or more wells; An inflow passage and an outflow passage are disposed on the side surface of the well, The inflow passage is disposed at an angle of 0 degree to 45 degrees with respect to a tangential direction or a side direction of the outer edge of the well, The liquid outlet at the upstream end of the outflow passage is disposed at an angle of 240 ° to 315 ° in the sample flow direction from the line connecting the liquid inlet at the downstream end of the inflow passage and the center of the well.
  • a method for analyzing a liquid sample using a microchip The microchip has one or more wells; An inflow passage and an outflow passage are disposed on the side surface of the well, The inflow passage is disposed at an angle of 0 degree to 45 degrees with respect to a tangential direction or a side direction of the outer edge of the well, The liquid outlet at the upstream end of the outflow passage is disposed at an angle of 240 ° to 315 ° in the sample flow direction from the line connecting the liquid inlet at the downstream end of the inflow passage and the center of the well.

Abstract

La présente invention concerne une micropuce dans laquelle l'occurrence d'un vide dans un puits lorsqu'un liquide s'est écoulé est inhibée. Cette micropuce dans laquelle s'écoule un liquide comporte un ou plusieurs puits, avec un passage d'écoulement d'entrée et un passage d'écoulement de sortie étant disposés dans la surface latérale du puits, le passage d'écoulement d'entrée étant agencé selon un angle de 0 à 45° par rapport à une direction tangentielle du bord externe ou une direction latérale du puits, et l'orifice d'écoulement de sortie de liquide au niveau de la partie d'extrémité amont du passage d'écoulement de sortie étant agencé à une position avec un angle de 240 à 315° par rapport à la direction d'écoulement d'un liquide par rapport à un segment de ligne reliant l'oxygène d'écoulement d'entrée de liquide au niveau de la partie d'extrémité aval du passage d'écoulement d'entrée et le centre du puits.
PCT/JP2016/050912 2015-03-09 2016-01-14 Micropuce, puits de micropuce, dispositif d'analyse utilisant une micropuce, et procédé d'analyse utilisant une micropuce WO2016143377A1 (fr)

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JP2015045510 2015-03-09
JP2015-045510 2015-03-09

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WO2016143377A1 true WO2016143377A1 (fr) 2016-09-15

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070280857A1 (en) * 2006-06-02 2007-12-06 Applera Corporation Devices and Methods for Positioning Dried Reagent In Microfluidic Devices
JP2012523829A (ja) * 2009-04-15 2012-10-11 コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ 気体を含まない流体チャンバ
JP2014030411A (ja) * 2012-08-06 2014-02-20 Sony Corp 核酸増幅反応用マイクロチップ

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070280857A1 (en) * 2006-06-02 2007-12-06 Applera Corporation Devices and Methods for Positioning Dried Reagent In Microfluidic Devices
JP2012523829A (ja) * 2009-04-15 2012-10-11 コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ 気体を含まない流体チャンバ
JP2014030411A (ja) * 2012-08-06 2014-02-20 Sony Corp 核酸増幅反応用マイクロチップ

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