EP1383862A2 - Incubateur et procede d'incubation menageant l'organisme mis a incuber - Google Patents
Incubateur et procede d'incubation menageant l'organisme mis a incuberInfo
- Publication number
- EP1383862A2 EP1383862A2 EP02706840A EP02706840A EP1383862A2 EP 1383862 A2 EP1383862 A2 EP 1383862A2 EP 02706840 A EP02706840 A EP 02706840A EP 02706840 A EP02706840 A EP 02706840A EP 1383862 A2 EP1383862 A2 EP 1383862A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- well
- organism
- oven
- culture
- wells
- Prior art date
- Legal status (The legal status 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 status listed.)
- Withdrawn
Links
Classifications
-
- 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/50—Means for positioning or orientating the apparatus
-
- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/14—Incubators; Climatic chambers
-
- 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
- C12M41/48—Automatic or computerized control
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
Definitions
- the present invention relates to incubators and methods of incubating organisms. It is particularly applicable for putting embryos in culture and monitoring them under the best possible temperature and atmospheric conditions of development.
- sperm and eggs are put together. After this fertilization by conventional in vitro fertilization or micro-injection, the gametes are placed in culture media and stored in an incubator. They are taken out daily, once or twice a day, for a change of environment, observation under a microscope, or any other manipulation, until the transfer in utero of the embryo.
- the invention remedies these drawbacks by an incubator and by an incubation method which eliminates thermal shock and the risk of bacterial and mycological contamination while facilitating the change of environments and automating it so as to reduce handling errors. and traumatic manipulation.
- the invention used in the context of in vitro fertilization therefore makes it possible to improve the number of viable and good quality embryos.
- organism means not only the gametes (oocytes and spermatozoa), the fertilized oocytes, the embryonic cultures and the embryos but also any cell culture such as stem cells, pathogenic or non-pathogenic microorganisms , etc ....
- the subject of the invention is therefore an incubator comprising a sealed oven equipped with a door, a culture box with several wells placed in the oven, an organism being in one of the wells of said box, characterized in that it comprises a controlled means of the exterior, intended to exit from the well in which the organism is located, to put it in another well filled with a new culture medium.
- the means, controlled from the outside, intended to exit from the well the organism to put it in another well filled with a culture medium is a pipette.
- the microscope is mounted inside or outside the oven so as to observe the culture of the organism and to monitor the various manipulations (transfer of the organism from a well to the 'other and filling of wells), the microscope being connected to a display screen placed outside the oven.
- the microscope stage is located inside the oven.
- a protective case may be necessary to its operation under the conditions of temperature and concentration of N 2/2 and 0 in C0 2 within one oven.
- the microscope advantageously used in the context of the invention has a filter which protects the organism from the deleterious effects of light, when it is observed under a microscope.
- the incubator object of the present invention accepts variants with a microscope with a fixed or mobile stage and / or a microscope with a fixed or mobile axis.
- the incubator which is the subject of the present invention can be divided into zones and include at least: an "interventional zone" which is dedicated to the observation and to the various manipulations of the embryos (filling of media and changing of wells),
- the three different zones can be on the same level or on different levels.
- the incubator object of the present invention comprises means, controlled from the outside of the oven, for moving step by step in one oven culture dishes with several wells. Any means of stepwise movement of a culture dish can be used in the context of the present invention.
- the means of movement used consists of:
- the incubator may include a source of culture medium communicating with a valve conduit opening at the above a well and valve control means so as to pour into the well a prescribed quantity of the culture medium.
- the source of medium may be outside the incubator or may be provided therein, for example, in an area dedicated to the storage of consumables.
- the conduit opens out above the well located at said location, which again makes it possible to monitor the operations using the microscope.
- the conduit opens above the well just upstream of the well located at said location. It is thus possible to simultaneously fill this well upstream and remove the embryo from the well located at said location by aspirating it using the pipette. There is also more room to bring the conduit over the well.
- the incubator which is the subject of the present invention comprises means for displacement of the valve useful for filling medium from one well to another.
- the oven advantageously has conditions of temperature and concentration of N 2 , 0 2 and C 2 ideal for the development of cell cultures and, more particularly, embryonic cultures. These conditions are advantageously a temperature at 37 ° C and an atmosphere at 5% O 2 , 5% CQ, and 90% N 2 .
- the incubator which is the subject of the present invention is equipped with sensors allowing self-regulation and monitoring of the atmosphere inside the oven. Thanks to these sensors, it is possible to control, during the entire duration of the culture, the temperature and the concentration of N 2 , of 0 2 and of C0 2 . Any sensor capable of measuring variations in temperature and concentration of N 2 , 0 2 and C0 2 , under the conditions of the oven, can be used in the context of the present invention.
- the invention also relates to a method of incubating an organism, which consists in placing the organism in a sealed oven, in incubating it in a culture well filled with a culture medium and to change it n times the culture medium, n being an integer greater than 1 and less than 50, characterized in that it consists of i) putting the organism in the 'one of a series of (n + 1) wells from a box placed in the oven, while the n other wells of the series are free of any organism, ii) to be moved, without removing it from the oven , the organism of the well in which it is to put it in the next well of the series filled with a new medium and iii) to repeat the displacement at most as many times as there remain of available wells.
- All operations are carried out inside the incubator under the supervision of the microscope and can be controlled automatically by a computer.
- the process which is the subject of the present invention consists in observing under a microscope the color change in the culture medium containing an indicator and contained in the well of a series of (n + 1) colored wells in which the organism and to change the well organism during this turn.
- a colored indicator in particular of pH, for example phenol red
- FIG. 1 represents a schematic sectional view of an incubator according to invention
- FIG. 1 The schematic sectional view of one incubator according to the invention, shown in Figure 1 shows that it comprises a sealed oven 1 provided with a door 2 which can be closed in a sealed manner. Inside the oven 1 is mounted an endless conveyor belt 3 driven by a motor roller 4 and passing over a return roller 5. The motor roller 4 is rotated by a motor 6 located outside of the oven 1, but can also be provided inside thereof, and which, in in this case, is connected to the motor roller 4 by a line 7. The stepping motor 6 is controlled by a computer 8 via a control line 9. On the upper strand of the conveyor 3, a culture dish 10 is placed comprising three wells 11a, 11b and 11e. The well 11a is upstream from the well 11b in the direction of movement F of the conveyor, while the well 11a is downstream.
- the box 10 is held on the conveyor 3 by buttresses 12 which prevent it from moving inadvertently.
- a fixed microscope 13 is mounted which is connected by a line 14 to a display device 15 making it possible to see the well 11b located at this location.
- a pipette 16 controlled by the computer 8 via a line 17.
- the oven 2 is carried a bottle 21 of culture medium which, by a conduit 22 provided with a valve 23, opens above the well 11a or, according to a variant which is also shown in the figure, above the well 11b on the understanding that one of these variants is used, but not both at the same time.
- An optical densitometer 24 with threshold striking makes it possible to follow the optical density of the culture medium of well 11b.
- the door 2 of the oven 1 is opened and it is placed in the linked well, filled with culture medium, in the oven 1, while the wells 11a and 11b are empty, being understood that they could also be already filled with culture medium.
- Door 2 is closed. It will not be opened again until cultivation is complete.
- the embryo is allowed to grow in the linked well which is then below the microscope 13 with the possibility, thanks to this microscope and possibly of the densitometer 24, of observing what is happening there and, if necessary, of deciding on the best time to change the culture medium.
- the pipette 16 is lowered into the linked well, the embryo is passed from the linked well into the pipette 16, the pipette 16 is removed from the linked well and the conveyor 3 is advanced by one step to bring now the well 11b opposite the microscope 13 at the location where the linked well was previously located. The pipette 16 is then lowered into the well 11b to obtain the position shown in the figure. The well 11b is filled with culture medium using the bottle 21 of the conduit 22 and the valve 23, if this operation had not already been carried out before, then the pipette 16 is lowered and the mixture is released.
- Figures 2 and 3 show two types of incubators capable of each containing several culture dishes.
- the incubator in Figure 3 is a system consisting of two stages. On the first (El), the dishes are stored, this stage therefore corresponding to the incubation zone (A). The tray is fixed. An airlock allows the introduction of 4 boxes simultaneously. The second floor
- (E2) is the intervention stage to which we pass via a manipulator arm.
- the axis of the microscope is fixed and the microscope stage on which the box to be observed is placed moves in x and in y, the microscope moving in z for focusing.
- the consumables In the center of the tray (C) are stored the consumables and the operations for removing media and cones and removing them are carried out.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Sustainable Development (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Clinical Laboratory Science (AREA)
- Thermal Sciences (AREA)
- General Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Computer Hardware Design (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0101779 | 2001-02-09 | ||
FR0101779A FR2820756B1 (fr) | 2001-02-09 | 2001-02-09 | Incubateur et procede d'incubation menageant l'organisme mis a incuber |
PCT/FR2002/000511 WO2002064728A2 (fr) | 2001-02-09 | 2002-02-11 | Incubateur et procede d'incubation menageant l'organisme mis a incuber |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1383862A2 true EP1383862A2 (fr) | 2004-01-28 |
Family
ID=8859829
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP02706840A Withdrawn EP1383862A2 (fr) | 2001-02-09 | 2002-02-11 | Incubateur et procede d'incubation menageant l'organisme mis a incuber |
Country Status (5)
Country | Link |
---|---|
US (1) | US20040064013A1 (ja) |
EP (1) | EP1383862A2 (ja) |
JP (1) | JP2004528020A (ja) |
FR (1) | FR2820756B1 (ja) |
WO (1) | WO2002064728A2 (ja) |
Families Citing this family (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB0219779D0 (en) * | 2002-08-23 | 2002-10-02 | Smiths Group Plc | Embryo transfer catheters |
DE20214480U1 (de) | 2002-09-18 | 2003-01-02 | Europäisches Laboratorium für Molekularbiologie, 69117 Heidelberg | Vorrichtung zur Klimakontrolle |
IL154677A0 (en) * | 2003-02-27 | 2003-09-17 | Univ Bar Ilan | A method and apparatus for manipulating an individual cell |
WO2004109361A1 (ja) * | 2003-06-02 | 2004-12-16 | Nikon Corporation | 顕微鏡装置 |
US9200245B2 (en) | 2003-06-26 | 2015-12-01 | Seng Enterprises Ltd. | Multiwell plate |
WO2005028612A1 (ja) * | 2003-09-22 | 2005-03-31 | Hirata Corporation | 細胞観察チェンバー内の溶液温度調整装置 |
DE10344284A1 (de) * | 2003-09-24 | 2005-05-04 | Keyneurotek Ag | Vorrichtung und Verfahren zur automatisierten Durchführung von Laborarbeitsschritten |
FI117899B (fi) * | 2004-07-09 | 2007-04-13 | Chip Man Technologies Oy | Solujen kasvatusalusta ja sen käyttö |
FI116946B (fi) * | 2004-07-09 | 2006-04-13 | Chip Man Technologies Oy | Laitteisto solujen kuvaamiseksi |
JP5010867B2 (ja) * | 2005-09-22 | 2012-08-29 | オリンパス株式会社 | 培養顕微鏡装置 |
JP4821279B2 (ja) | 2005-11-11 | 2011-11-24 | 株式会社ニコン | 培養装置 |
WO2009003487A2 (en) | 2007-06-29 | 2009-01-08 | Unisense Fertilitech A/S | A device, a system and a method for monitoring and/or culturing of microscopic objects |
US9975118B2 (en) | 2007-11-15 | 2018-05-22 | Seng Enterprises Ltd. | Device for the study of living cells |
US9145540B1 (en) | 2007-11-15 | 2015-09-29 | Seng Enterprises Ltd. | Device for the study of living cells |
CN102083954B (zh) * | 2008-07-05 | 2014-09-03 | 尤尼森斯繁殖技术公司 | 一对一识别系统 |
GB2461707B (en) * | 2008-07-08 | 2013-06-26 | Ruskinn Life Sciences Ltd | Laboratory apparatus for a controlled environment |
US11034927B2 (en) * | 2015-03-31 | 2021-06-15 | Thrive Bioscience, Inc. | Cell culture incubators with integrated imaging systems |
CN107418888A (zh) * | 2017-09-26 | 2017-12-01 | 上海莫杜生物科技有限公司 | 一种微生物循环接种装置及其接种方法 |
CN108728333A (zh) * | 2018-04-28 | 2018-11-02 | 广州滨鑫生物科技有限公司 | 一种生物技术用细胞培养箱及其培养方法 |
KR102475749B1 (ko) * | 2021-12-27 | 2022-12-08 | 전태진 | 무인 이동식 혐기성 미생물 배양장치 |
US20240141279A1 (en) * | 2022-10-31 | 2024-05-02 | Power to Innovate Technologies Inc. | Incubation monitoring apparatus |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BE627235A (ja) * | 1962-02-02 | |||
US4892830A (en) * | 1987-04-02 | 1990-01-09 | Baylor College Of Medicine | Environmentally controlled in vitro incubator |
ATE154981T1 (de) * | 1990-04-06 | 1997-07-15 | Perkin Elmer Corp | Automatisiertes labor für molekularbiologie |
AU7909091A (en) * | 1990-05-16 | 1991-12-10 | Scientific Imaging Instruments, Inc. | Method and apparatus for preparing a liquid specimen |
US5083558A (en) * | 1990-11-06 | 1992-01-28 | Thomas William R | Mobile surgical compartment with micro filtered laminar air flow |
US5192506A (en) * | 1991-02-14 | 1993-03-09 | P B Diagnostic Systems, Inc. | Incubator port closure for automated assay system |
JP3067347B2 (ja) * | 1991-10-30 | 2000-07-17 | 株式会社島津製作所 | ゲル状ビーズの選別装置 |
AU656826B2 (en) * | 1991-10-31 | 1995-02-16 | Microscan, Inc. | Specimen processing and analyzing systems with associated fluid dispensing apparatus |
EP0823483A4 (en) * | 1996-01-24 | 2001-04-18 | Matsushita Electric Ind Co Ltd | METHOD FOR MEASURING PHYSICO-CHEMICAL PROPERTIES OF TISSUES OR CELLS, METHOD FOR EXAMINING CHEMICAL SUBSTANCES AND EQUIPMENT THEREOF |
US6008010A (en) * | 1996-11-01 | 1999-12-28 | University Of Pittsburgh | Method and apparatus for holding cells |
US6166761A (en) * | 1997-01-09 | 2000-12-26 | Interface Multigrad Technology | Method and apparatus for monitoring a biological sample |
DE69836736T2 (de) * | 1997-05-01 | 2007-10-04 | Sophion Bioscience A/S | Vorrichtung zur automatischen elektrodenpositionierung |
-
2001
- 2001-02-09 FR FR0101779A patent/FR2820756B1/fr not_active Expired - Fee Related
-
2002
- 2002-02-11 EP EP02706840A patent/EP1383862A2/fr not_active Withdrawn
- 2002-02-11 WO PCT/FR2002/000511 patent/WO2002064728A2/fr not_active Application Discontinuation
- 2002-02-11 JP JP2002565043A patent/JP2004528020A/ja active Pending
-
2003
- 2003-08-06 US US10/635,278 patent/US20040064013A1/en not_active Abandoned
Non-Patent Citations (1)
Title |
---|
See references of WO02064728A2 * |
Also Published As
Publication number | Publication date |
---|---|
FR2820756B1 (fr) | 2004-01-23 |
WO2002064728A3 (fr) | 2003-11-06 |
WO2002064728A2 (fr) | 2002-08-22 |
US20040064013A1 (en) | 2004-04-01 |
FR2820756A1 (fr) | 2002-08-16 |
JP2004528020A (ja) | 2004-09-16 |
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