WO2014091524A1 - Dispositif de collecte d'objet - Google Patents

Dispositif de collecte d'objet Download PDF

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Publication number
WO2014091524A1
WO2014091524A1 PCT/JP2012/007973 JP2012007973W WO2014091524A1 WO 2014091524 A1 WO2014091524 A1 WO 2014091524A1 JP 2012007973 W JP2012007973 W JP 2012007973W WO 2014091524 A1 WO2014091524 A1 WO 2014091524A1
Authority
WO
WIPO (PCT)
Prior art keywords
suction
tip
well
main body
cell aggregate
Prior art date
Application number
PCT/JP2012/007973
Other languages
English (en)
Japanese (ja)
Inventor
伊藤　三郎
Original Assignee
ヤマハ発動機株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ヤマハ発動機株式会社 filed Critical ヤマハ発動機株式会社
Priority to JP2014551740A priority Critical patent/JP5902319B2/ja
Priority to PCT/JP2012/007973 priority patent/WO2014091524A1/fr
Publication of WO2014091524A1 publication Critical patent/WO2014091524A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/12—Well or multiwell plates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements

Definitions

  • the present invention relates to an object recovery apparatus used when recovering an object such as a cell while observing the object.
  • FIG. 16 and FIG. 17 are schematic diagrams for explaining the configuration of a conventional object recovery apparatus.
  • the moving device M driving unit
  • suction pipette P suction means
  • the suction pipette P suction means
  • FIG. 16 reference symbol A1 indicates the irradiation direction of the irradiation light. In this case, in order to recover the cell AG on the stage S (in FIG.
  • FIG. 1 is a schematic diagram illustrating the configuration of an object recovery apparatus 1 according to this embodiment.
  • the object recovery apparatus 1 of the present embodiment is a stage on which a well plate 4 including a well 3 that holds a cell aggregate 2 to be sucked (spheroid spheroid, object, see FIG. 2B described later) is placed. 5 and a capacitor 6 (irradiation means) which is arranged spaced apart upward from the well plate 4 placed on the stage 5 and irradiates the cell aggregate 2 held in the well 3 with irradiation light from above.
  • the imaging device 7 (observation means) that is arranged below the well plate 4 placed on the stage 5 and that observes the cell aggregate 2 held in the well 3 from below, and the cells held in the well 3
  • a suction pipette 9 suction means for generating a suction force for suction
  • a moving device 10 drive means for moving the suction pipette 9 up and down Equipped with a.
  • the capacitor 6 and the imaging device 7 are devices that constitute an illumination system and an imaging system of an inverted phase contrast microscope, respectively.
  • the cell aggregate 2 (target object) is a sample recovered using the target object recovery apparatus 1 of the present embodiment.
  • the cell aggregate 2 is normally held in the well 3 in a state of being contained in a liquid 11 such as a cell culture solution or a cell treatment solution in order to prevent deterioration due to drying or the like (see FIG. 2B). ).
  • a liquid 11 such as a cell culture solution or a cell treatment solution in order to prevent deterioration due to drying or the like (see FIG. 2B).
  • impurities such as organelles, cells that do not form aggregates, and the like may be removed in advance so that the imaging apparatus 7 to be described later can easily identify the cell aggregates 2 to be aspirated.
  • the method of removing these is not particularly limited, and a method of removing the cell culture solution containing the cell aggregate 2 containing impurities through a filter or the like can be employed. By passing through the filter, a liquid 11 such as a cell culture solution mainly containing a cell aggregate 2 having a specific size and
  • the distance between the capacitor 6 and the stage 5 is limited to a predetermined distance when the Kohler illumination system is employed. Therefore, when the height of the well plate 4 is too large, when the tip 83 of the suction tip 8 is moved into and out of the well 3 by moving the suction pipette 9 up and down by the moving device 10 described later, the tip 83 and the capacitor 6 may interfere with each other, which may hinder accurate operation. Therefore, it is preferable that the height of the well plate 4 is such that a space is formed between the capacitor 6 and the upper surface 41 of the well plate 4 so as not to interfere with the above operation.
  • the tip portion 83 is disposed above the well 3, and the length of the tip portion 83 is further between the bent portion 81 and the capacitor 6 immediately before being inserted into the well 3.
  • the height is such that a certain amount of space is created. In this case, even if the tip portion 83 is not inserted into the well 3, a space is formed above the bent portion 81 by the length of the tip portion 83, so that interference between the tip portion 83 and the capacitor 6 is prevented. It is prevented.
  • thermoplastic resin a thermosetting resin, a photocurable resin etc.
  • the light-transmitting material polyethylene resin; polyethylene naphthalate resin; polypropylene resin; polyimide resin; polyvinyl chloride resin; cycloolefin copolymer; norbornene resin-containing polyether sulfone resin; Aromatic polyamide resin; (meth) acrylic resin such as poly (meth) methyl acrylate; styrene resin such as polystyrene and styrene-acrylonitrile copolymer; polycarbonate resin; polyester resin; phenoxy resin; butyral resin; Cellulose resins such as cellulose acetate and cellulose acetate butyrate; epoxy resins; phenol resins; silicone resins; Also, inorganic materials such as metal alkoxides, ceramic precursor polymers, solutions obtained by hydrolytic polymerization of
  • the size of the well 3 (the diameter of the opening) is not particularly limited as long as it can hold at least a part of the cell aggregate 2 to be collected (in FIG. 2), the diameter of the opening is larger than the diameter of the cell aggregate 2 so that the entirety of 2 fits in the well 3.
  • the depth of the well 3 is not particularly limited, and may be a depth that can hold the cell aggregate 2 in the well 3 and may be shorter than the length of the distal end portion 83 of the suction tip 8 described later.
  • a well 3 having an opening with a diameter of 9 mm and a depth of 11 mm is illustrated, and a spherical cell aggregate having a diameter of 80 ⁇ m is held in the inner bottom 31 of the well 3.
  • FIG. 3 is a schematic diagram illustrating the suction tip 8 and the suction pipette 9 of the present embodiment.
  • the suction pipette 9 includes a tubular main body 91, an opening 92 at one end of the main body 91 is connected to a connection port 86 of the suction tip 8 and communicates with a tubular passage 84 of the suction tip 8, and an opening at the other end.
  • the part is connected to a syringe pump (not shown).
  • the syringe pump generates suction force in the tubular main body portion 91 and the tubular passage 84 of the suction tip 8 communicating with the main body portion 91.
  • the size (internal volume) of the suction tip 8 is not particularly limited, and can be about 1 ⁇ L to 5 mL, for example, depending on the application.
  • the internal volume of the suction tip 8 of this embodiment is 20 ⁇ L.
  • the material constituting the suction tip 8 is not particularly limited as long as it is a material used for a normal suction tip.
  • a resin such as polypropylene or polystyrene can be adopted as the material.
  • the suction tip 8 of this embodiment is made of polypropylene.
  • the diameter of the suction port 82 and the inner diameter of the tubular passage 84 are not particularly limited as long as the cell aggregate 2 can be sucked.
  • the inner diameter of the suction port 82 of the suction tip 8 of this embodiment is 150 ⁇ m, and the inner diameter of the tubular passage 84 is 4500 ⁇ m (maximum).
  • the tubular member 12a includes one end opening 121a communicating with the tubular passage 84b and the other end opening 122a functioning as a suction port for sucking the cell aggregate 2.
  • a hydrophobic coat layer made of an acrylic silicone resin is provided on the inner peripheral wall of the tubular member 12a.
  • the main body portion 85b and the tubular member 12a are connected at the connection portion 13a.
  • the suction tip 8b of the present embodiment is not formed with an L-shaped bent portion in the main body portion 85b, but is a straight shape, and the suction tip 8a described with reference to FIG. 4 in the second embodiment. Because of this, duplicate description is omitted.
  • connection portion between the main body portion and the tubular member has an inner peripheral wall of the tubular passage in a state where an end portion including the one end opening portion of the tubular member is inserted into the tubular passage of the main body portion. It is preferable that the connecting portion is formed by thermally shrinking the main body portion so as to be crimped to the outer peripheral wall of the end portion.
  • a hydrophobic coat layer on the inner peripheral wall of the tubular member, for example, when collecting an object contained in a liquid such as a cell culture solution, capillary action that occurs when the suction port comes into contact with the liquid. The influence can be suppressed, and the resistance when collecting the object contained in the liquid can be reduced. In addition, the liquid or cell agglomerates sucked at the time of discharge are easily discharged and hardly remain in the tubular passage or the tubular member.
  • the object is preferably a living cell.
  • a cell-derived aggregate derived from a living body has a living body-like environment in which the interaction between cells is taken into consideration inside the cell aggregate, rather than the test results obtained using a single cell. Results are taken into account, and the experimental conditions can be adjusted to the conditions that are more suitable for the environment in the living body, which is important in the field of regenerative medicine and the development of pharmaceuticals such as anticancer drugs. ing.
  • cell aggregates have various shapes. In the present invention, since only cell aggregates suitable for various tests can be collected while observing the shape, the shape of the cell aggregates used for the test can be aligned. Reliable results can be obtained in the field of regenerative medicine and development of pharmaceuticals such as anticancer agents.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Clinical Laboratory Science (AREA)
  • Analytical Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne un dispositif de collecte d'objet équipé : d'un support sur lequel est posée une plaque à puits ; d'un moyen d'irradiation disposé au-dessus de la plaque à puits et conçu pour envoyer de la lumière ; d'un moyen d'observation disposé sous la plaque à puits ; d'un tube d'aspiration pour aspirer un objet ; d'un moyen d'aspiration pour produire une force d'aspiration pour l'aspiration ; et d'un moyen d'entraînement pour déplacer le moyen d'aspiration verticalement. Le tube d'aspiration présente une courbure en forme de L et est pourvu d'un corps principal et d'un orifice d'aspiration. Le moyen d'aspiration insère la partie avant de la courbure dans un puits dans un sens sensiblement vertical et dispose l'extrémité arrière en position horizontale s'étendant latéralement de l'espace séparant le moyen d'irradiation et la plaque à puits. Le moyen d'entraînement déplace le moyen d'aspiration verticalement tout en maintenant la position horizontale. La présente invention permet de collecter un objet en déplaçant l'extrémité avant du tube d'aspiration verticalement et en la mettant près de l'objet tout en observant ce dernier sous une irradiation de lumière adéquate.
PCT/JP2012/007973 2012-12-13 2012-12-13 Dispositif de collecte d'objet WO2014091524A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2014551740A JP5902319B2 (ja) 2012-12-13 2012-12-13 対象物回収装置
PCT/JP2012/007973 WO2014091524A1 (fr) 2012-12-13 2012-12-13 Dispositif de collecte d'objet

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2012/007973 WO2014091524A1 (fr) 2012-12-13 2012-12-13 Dispositif de collecte d'objet

Publications (1)

Publication Number Publication Date
WO2014091524A1 true WO2014091524A1 (fr) 2014-06-19

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JP (1) JP5902319B2 (fr)
WO (1) WO2014091524A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016116460A (ja) * 2014-12-19 2016-06-30 パナソニック株式会社 細胞培養装置
CN105758667A (zh) * 2016-04-19 2016-07-13 山东出入境检验检疫局检验检疫技术中心 恒定面积浆板取样器及取样方法
WO2017110005A1 (fr) * 2015-12-25 2017-06-29 ヤマハ発動機株式会社 Procédé de prélèvement d'objet cible
LU100170B1 (de) * 2017-04-13 2018-10-15 Cytena Gmbh Verfahren zum Prozessieren einer flüssigen Probe
JPWO2019176093A1 (ja) * 2018-03-16 2021-01-07 株式会社島津製作所 細胞ピッキング装置

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP7459064B2 (ja) * 2018-09-11 2024-04-01 コーニング インコーポレイテッド ピペット構造体及びその利用方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006349502A (ja) * 2005-06-16 2006-12-28 Aloka Co Ltd 自動分注装置及び自動分注方法
JP2008520978A (ja) * 2004-11-22 2008-06-19 エフ ホフマン−ラ ロッシュ アクチェン ゲゼルシャフト 湾曲微細構造
JP2009504161A (ja) * 2005-08-11 2009-02-05 バイオトローブ インコーポレイティッド アッセイ、合成及び保管のための装置、並びにその製造法、使用法及び操作法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3388825B2 (ja) * 1993-08-25 2003-03-24 株式会社スタックシステム 分注装置
JPH07239336A (ja) * 1994-02-25 1995-09-12 Olympus Optical Co Ltd 医療用分析機

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008520978A (ja) * 2004-11-22 2008-06-19 エフ ホフマン−ラ ロッシュ アクチェン ゲゼルシャフト 湾曲微細構造
JP2006349502A (ja) * 2005-06-16 2006-12-28 Aloka Co Ltd 自動分注装置及び自動分注方法
JP2009504161A (ja) * 2005-08-11 2009-02-05 バイオトローブ インコーポレイティッド アッセイ、合成及び保管のための装置、並びにその製造法、使用法及び操作法

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016116460A (ja) * 2014-12-19 2016-06-30 パナソニック株式会社 細胞培養装置
WO2017110005A1 (fr) * 2015-12-25 2017-06-29 ヤマハ発動機株式会社 Procédé de prélèvement d'objet cible
CN105758667A (zh) * 2016-04-19 2016-07-13 山东出入境检验检疫局检验检疫技术中心 恒定面积浆板取样器及取样方法
CN105758667B (zh) * 2016-04-19 2018-05-18 山东出入境检验检疫局检验检疫技术中心 恒定面积浆板取样器及取样方法
LU100170B1 (de) * 2017-04-13 2018-10-15 Cytena Gmbh Verfahren zum Prozessieren einer flüssigen Probe
US11845044B2 (en) 2017-04-13 2023-12-19 cytena Bioprocess Solutions co., Ltd. Method for processing a liquid sample
JPWO2019176093A1 (ja) * 2018-03-16 2021-01-07 株式会社島津製作所 細胞ピッキング装置

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JPWO2014091524A1 (ja) 2017-01-05
JP5902319B2 (ja) 2016-04-13

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