CN116536155A - Chip culture device and application method thereof - Google Patents

Chip culture device and application method thereof Download PDF

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Publication number
CN116536155A
CN116536155A CN202310555267.6A CN202310555267A CN116536155A CN 116536155 A CN116536155 A CN 116536155A CN 202310555267 A CN202310555267 A CN 202310555267A CN 116536155 A CN116536155 A CN 116536155A
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CN
China
Prior art keywords
chip
culture
incubator
negative pressure
lifting
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Granted
Application number
CN202310555267.6A
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Chinese (zh)
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CN116536155B (en
Inventor
尹彬沣
朱浩宇
曾施宇
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Yangzhou University
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Yangzhou University
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Priority to CN202310555267.6A priority Critical patent/CN116536155B/en
Publication of CN116536155A publication Critical patent/CN116536155A/en
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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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/48—Holding appliances; Racks; Supports
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
    • B01L3/0217—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids of the plunger pump type
    • B01L3/0234—Repeating pipettes, i.e. for dispensing multiple doses from a single charge
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/0289—Apparatus for withdrawing or distributing predetermined quantities of fluid
    • B01L3/0293—Apparatus for withdrawing or distributing predetermined quantities of fluid for liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • 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
    • C12M23/16—Microfluidic devices; Capillary tubes
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/067—Hepatocytes
    • C12N5/0671—Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2503/00—Use of cells in diagnostics
    • C12N2503/02—Drug screening
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/10—Screening for compounds of potential therapeutic value involving cells
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
    • Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures

Abstract

The invention discloses a chip culture device and a use method thereof, the chip culture device comprises an incubator with a containing cavity, a supporting plate is fixed in the incubator, the supporting plate is connected with a culture plate, a plurality of groups of chip components are distributed on the front and back directions of the culture plate, the chip components are composed of a plurality of chips which are arranged at intervals in the left and right directions, a movable seat capable of reciprocating rectilinear motion is connected in the incubator, a plurality of injectors capable of lifting are distributed at the lower end of the movable seat, a multichannel peristaltic pump and a liquid storage shell are fixedly connected at the outer side of the incubator, a plurality of groups of mutually independent liquid storage parts are arranged in the liquid storage shell, each liquid storage part comprises a plurality of mutually independent liquid storage tanks, a plurality of channel input ends of the multichannel peristaltic pump respectively correspond to the plurality of liquid storage parts one by one, a plurality of channel output ends of the multichannel peristaltic pump respectively correspond to the plurality of injectors one by one, and the multichannel peristaltic pump can pump solution in the liquid storage parts into the injectors in sequence; the invention can simultaneously culture a plurality of chips in a sample adding way, and improves the culture efficiency.

Description

Chip culture device and application method thereof
Technical Field
The invention relates to the technical field of chip culture, in particular to a chip culture device and a use method thereof.
Background
Organ chip is a popular research direction in the vicinity of the current microfluidic technology. The organ chip is a multi-channel three-dimensional cell culture device comprising a continuous perfusion chamber, can truly reflect the intracellular environment of a human body, and has a good application prospect in the aspect of drug screening.
In the prior art, a Chinese patent with the name of 'a micro-vessel liver chip based on cell aggregate' and the application number of 2015107205789 is disclosed, wherein the Chinese patent comprises a cell aggregate enrichment region and a plurality of cell co-culture regions, and a culture system inlet and a culture system outlet are respectively arranged at two ends of the Chinese patent, so that the preparation of a liver disease model and the research of pharmacokinetics and pharmaceutical activity can be carried out; however, it can only perform cell culture on a single chip, but cannot perform cell culture on a plurality of chips at the same time, and the chip culture efficiency is low.
Disclosure of Invention
This section is intended to outline some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section as well as in the description summary and in the title of the application, to avoid obscuring the purpose of this section, the description summary and the title of the invention, which should not be used to limit the scope of the invention.
The present invention has been made in view of the above and/or problems occurring in the conventional chip culture.
Therefore, the invention aims to solve the problem that a plurality of chips cannot be injected in the prior art, promote the mixing of different solutions and improve the reaction efficiency.
In order to solve the technical problems, the invention provides the following technical scheme: the utility model provides a chip culture device, its includes the incubator that has the holding chamber, incubator internal fixation has the backup pad, swing joint has the culture plate in the backup pad, a plurality of groups of chip components that wait to cultivate have been arranged in the front and back direction of culture plate, the chip components comprises a plurality of chips that set up in the left and right directions interval, incubator internal connection above the culture plate has the movable seat that can do reciprocal rectilinear motion, the lower extreme of movable seat has arranged a plurality of syringes that can go up and down, incubator outside fixedly connected with multichannel peristaltic pump and stock solution casing, have a plurality of mutually independent stock solution portions of group in the stock solution casing, the stock solution portion includes a plurality of mutually independent stock solution groove, a plurality of channel inputs of multichannel peristaltic pump respectively with a plurality of group stock solution portion one-to-one, a plurality of channel outputs of multichannel peristaltic pump respectively with a plurality of syringe one-to-one, multichannel peristaltic pump can pump the solution in the stock solution portion into the syringe in proper order.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the liquid outlet main pipe is connected with a plurality of liquid outlet branch pipes which are in one-to-one correspondence with liquid storage tanks in the same group of liquid storage parts, a liquid outlet one-way valve is connected on the liquid outlet branch pipes, and the liquid outlet main pipe is connected with corresponding channel input ends.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the incubator is connected with a lifting seat which can do reciprocating rectilinear motion in the front-back direction and can lift, a plurality of air inlet devices which correspond to chips in any group of chip components one by one are arranged on the lower side of the lifting seat, a total air pipe is fixedly connected below the lifting seat, the output end of the air inlet device is connected with an air inlet valve, the output end of the air inlet valve is connected with an air distribution pipe, the upper end of the chip is provided with a composite air hole, the lower side of the tail end of the air distribution pipe is fixedly provided with an inserting pipe which corresponds to the composite air hole, one side of the air distribution pipe, which is far away from the inserting pipe, is connected to the total air pipe, and an air suction one-way valve is connected to the air distribution pipe between the inserting pipe and the total air pipe.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the side end of the main air pipe is connected with a negative pressure air pipe, a negative pressure intubation tube which is vertically arranged and corresponds to the air suction hole is fixed on the negative pressure air pipe, and the negative pressure intubation tube can be just inserted into the culture plate through the corresponding air suction hole.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the incubator of backup pad below internal fixedly connected with pipe connection driver, be connected with the telescopic link that can reciprocate rectilinear motion in the direction of height on the pipe connection driver, the upper end of telescopic link is connected with the joint, be connected with the negative pressure connecting pipe on the joint, the negative pressure connecting pipe can just be inserted in the culture plate through the negative pressure connecting hole, and the negative pressure connecting pipe is kept away from the one end and the negative pressure equipment connection of joint.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the incubator is characterized in that a lower adjusting component used for adjusting the position of the lifting seat is further connected in the incubator, the lower adjusting component comprises a transverse frame fixedly connected to the inner side of the incubator, a lower transmission screw rod which is horizontally arranged is rotatably connected to the transverse frame, a first sliding table which is connected to the transverse frame in a sliding mode is connected to the lower transmission screw rod in a threaded mode, a vertical stand is fixedly connected to one side of the first sliding table, which is vertically arranged, of the lifting seat, a second sliding table which is connected to the vertical stand in a sliding mode is rotatably connected to the lifting screw rod, the lifting seat is fixedly connected to the second sliding table, a lifting motor is fixedly connected to the upper side of the vertical stand, the lifting motor is connected to the lifting screw rod, a lower transmission motor is fixedly connected to one end of the transverse frame in the front-rear direction, and the lower transmission motor is connected to the lower transmission screw rod.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the incubator of backup pad top is connected with the last adjusting part that is used for adjusting and removes the seat position, go up adjusting part including setting up in the place ahead of removing the seat and fixing the fixed plate inboard at the incubator, rotationally be connected with the transmission lead screw between fixed plate and the incubator, it is last on the transmission lead screw to remove seat threaded connection, be connected with the guide arm on the fixed plate of transmission lead screw left and right sides respectively, the guide arm passes to remove the seat and connects on the incubator, fixed plate front side fixedly connected with goes up the transmission motor, go up the transmission motor and go up the transmission lead screw connection.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the lower side of the movable seat is movably connected with a connecting plate capable of moving left and right, a plurality of lifting drivers are arranged on the lower side of the connecting plate, connecting rods capable of performing reciprocating linear motion in the height direction are connected to the lifting drivers, and the syringes are connected to the lower ends of the corresponding connecting rods.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the device is characterized by further comprising a rear containing box arranged at the rear side of the incubator, wherein a storage rack capable of lifting is connected in the rear containing box, a plurality of storage plates used for storing the culture plates are arranged in the height direction of the storage rack, and a push-pull assembly capable of pushing the culture plates to be placed on the support plates or pulling the culture plates to be placed on the storage plates is connected in the rear containing box of the culture plates.
As a preferable embodiment of the chip culture apparatus of the invention, wherein: the rear side fixedly connected with billet of culture plate, push-and-pull subassembly includes at least one fixed connection at the rearmounted straight line driver that holds the incasement and be at the supporter rear, be connected with the push-and-pull rod that can do reciprocal rectilinear movement in the fore-and-aft direction on the straight line driver, the end-to-end connection of push-and-pull rod has the electro-magnet, and when the electro-magnet was on, the billet was adsorbed on the electro-magnet, and the electro-magnet breaks away from the electro-magnet when the electro-magnet was cut off the power supply.
A method for culturing by using a chip culturing device comprises the following steps,
lifting a culture plate provided with a plurality of chips to be cultured to the height of a supporting plate, enabling a linear driver to act, enabling a push-pull rod to extend forwards, enabling an electromagnet to be attached to a steel bar, enabling the electromagnet to be electrified, and stopping the linear driver when the culture plate is pushed to a proper position on the supporting plate;
the position of the movable seat is regulated to enable the injector to be moved to the position of the forefront chip assembly, the position of the injector in the left-right direction is regulated to enable the injector to be injected to be aligned with the liquid inlet of the corresponding chip in sequence, the corresponding lifting driver acts to enable the injector to descend, when the injection needle of the injector is inserted into the chip through the corresponding liquid inlet, the lifting driver stops acting, the corresponding liquid outlet one-way valve is controlled to be opened according to the set liquid injection sequence, the multichannel peristaltic pump works to enable the solution in the corresponding channel to be pumped into the injector, and when the liquid inlet amount reaches the set liquid inlet amount threshold, the corresponding liquid outlet one-way valve and the multichannel peristaltic pump are closed, the lifting driver acts reversely to reset, and the lifting driver stops acting;
when the solution in the chip is required to be mixed after the injection at one stage is finished, the movable seat is moved backwards, the lower transmission motor acts, the lifting seat is moved forwards to a set position, the lifting seat is lowered, the plug-in pipe is inserted into the chip through the corresponding composite air hole, the negative pressure equipment works, the corresponding air suction check valve is controlled to be opened, the solution in the liquid storage tank of the chip flows to the direction of the reaction tank through the mixing flow passage in the chip, when the set first time threshold value is reached, the negative pressure equipment and the air suction check valve are closed, the air inlet pump is opened, the solution flows in the direction away from the reaction tank, when the set second time threshold value is reached, the air inlet pump is closed, when the mixing time reaches the set third time threshold value, the negative pressure equipment is opened, the solution enters the reaction tank, the negative pressure equipment is closed, the lifting seat is moved backwards, the movable seat moves forwards, the injection at the next stage is performed, and otherwise, the mixing step is returned;
after the liquid injection culture of one row of chip assemblies is finished, the movable seat moves back to the position of the next batch of chip assemblies, the steps are repeated until a plurality of groups of chip assemblies in the culture plate are subjected to the chip culture, and the next step is carried out;
the linear driver acts reversely to enable the push-pull rod to retract backwards, the culture plate is pulled to the corresponding storage plate, the electromagnet is powered off, and the culture plate is loosened.
The invention has the beneficial effects that: through the joint arrangement of the components such as the multichannel peristaltic pump, the liquid storage shell, the injector and the like, the sample adding of each chip in a group of chip components is realized; through joint setting of total trachea, minute trachea, bleed check valve, air inlet pump and admission valve, realize the independent circulation mixing of different on-chip solutions, improve the homogeneity that the solution mixes in the cultivation to improve later stage's cultivation effect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a front view of the present invention.
Fig. 2 is a first perspective view of the present invention with the door hidden.
Fig. 3 is a partial enlarged view at B in fig. 2.
Fig. 4 is a second perspective view of the present invention with the door hidden.
Fig. 5 is a partial enlarged view at C in fig. 4.
Fig. 6 is a partial enlarged view at D in fig. 4.
Fig. 7 is a view from A-A in fig. 1.
Fig. 8 is a perspective view of the present invention.
Fig. 9 is a partial enlarged view at E in fig. 8.
Fig. 10 is a three-dimensional structure diagram of the present invention after the door is hidden.
Fig. 11 is a partial enlarged view of F in fig. 10.
FIG. 12 is a perspective view showing a structure of a culture plate according to the present invention.
Fig. 13 is a perspective view of a hidden door of the present invention.
Fig. 14 is a partial enlarged view at G in fig. 13.
In the figure: 1 incubator, 101 fixed plate, 102 support plate, 2 reservoir housing, 201 reservoir, 201a reservoir, 3 multichannel peristaltic pump, 4 chamber door, 5 negative pressure device, 6 chip assembly, 601 chip, 7 lifting seat, 8 syringe, 9 lifting drive, 10 lower adjustment assembly, 1001 lifting motor, 1002 stand, 1003 lifting screw, 1004 second slipway, 1005 lower drive motor, 1006 lower drive screw, 1007 transverse frame, 1008 first slipway, 11 total air pipe, 1101 negative pressure air pipe, 1101a negative pressure cannula, 1102 air distribution pipe, 1102a insertion pipe, 12 air suction check valve, 13 air inlet valve, 14 air inlet device, 15 upper adjustment assembly, 1501 upper drive screw, 1502 upper drive motor, 1503 guide rod, 16 telescopic rod, 17 negative pressure connection pipe, 18 joint, 19 outlet header, 1901 outlet split pipe, 20 outlet check valve, 21 culture plate, 22 shelf, 2201 storage plate, 23 rod, 24 linear drive, 25 electromagnet, 26, 27 moving seat, 28 drive belt, 29 position adjustment seat, 30 position adjustment motor, 31 slide rail, 32 suction tank, 2, b, d composite suction port, d air inlet port, d air vent and d vent.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1
Referring to fig. 1 to 3, a first embodiment of the present invention provides a chip culturing apparatus capable of simultaneously performing sample culture on a plurality of chips 601 and improving culturing efficiency.
The utility model provides a chip culture device, including having incubator 1 that holds the chamber, incubator 1's front end is connected with the chamber door 4 that can open and shut, incubator 1 internal fixation has backup pad 102, swing joint has culture plate 21 in the backup pad 102, a plurality of installation sink d have been arranged to one side up of culture plate 21, a plurality of groups of chip subassembly 6 of waiting to cultivate have been arranged in the fore-and-aft direction of culture plate 21, chip subassembly 6 comprises a plurality of chips 601 that set up at the left-right direction in the interval, chip 601 just places on culture plate 21 through installation sink d, incubator 1 internal connection of culture plate 21 top has can be in the fore-and-aft direction in reciprocating rectilinear motion remove seat 27, a plurality of syringes 8 that can go up and down have been arranged to remove seat 27 downside and have been arranged a plurality of lift driver 9, be connected with the connecting rod that can be in the direction of height in reciprocating rectilinear motion, syringe 8 connection is in the lower extreme of corresponding connecting rod, incubator 1 outside fixedly connected with multichannel peristaltic pump 3 and stock solution reservoir 2, have a plurality of groups mutually independent portion of setting up and mutually independent portion of stock solution reservoir 2, a plurality of channels 201a are used for a plurality of channels 3 in the time of the peristaltic pump 201a, a plurality of the channel of respectively, a plurality of channel 201a are hidden in the time of the present multiple channel of the peristaltic pump 201a, a is realized in the multiple channel of the present application, a plurality of channel 201A is connected in the time, and a plurality of channel 201A are used for the channel of mutually independent channel 201, and the channel 201A is used in the channel is in the time, and the channel 201 is used in the channel is used in the time, and the channel is used for the channel, and the channel is used in the channel, and the channel is used.
Specifically, the liquid storage device further comprises a plurality of liquid outlet manifolds 19 which are in one-to-one correspondence with the liquid storage parts 201, a plurality of liquid outlet branch pipes 1901 which are in one-to-one correspondence with the liquid storage grooves 201a in the liquid storage parts 201 in the same group are connected to the liquid outlet manifolds 19, liquid outlet check valves 20 are connected to the liquid outlet branch pipes 1901, and the liquid outlet manifolds 19 are connected with corresponding channel input ends.
In this embodiment, the multi-channel peristaltic pump 3 is in the prior art, the liquid outlet main pipe 19 only gives a schematic diagram, the connection between the liquid outlet main pipe 19 and the multi-channel peristaltic pump 3 is in the prior art, not shown, the output ends of the channels of the multi-channel peristaltic pump 3 are connected with the corresponding syringes 8 through the pipelines, which is in the prior art, not shown, and the lifting driver 9 is preferably an electric push rod; in the initial state, each liquid outlet one-way valve 20 is in a closed state, and different liquid storage tanks 201a in the same group of liquid storage parts 201 are respectively filled with solutions required by culture; when the chip 601 is cultured, the corresponding liquid outlet one-way valve 20 is controlled to be opened according to the sample injection sequence required by culture, and when the liquid outlet one-way valve 20 is opened, the channel corresponding to the multi-channel peristaltic pump 3 connected with the liquid outlet one-way valve 20 is opened, and the multi-channel peristaltic pump 3 pumps the solution in the corresponding liquid storage tank 201a into the corresponding injector 8 so as to pump the solution into the liquid storage tank of the corresponding chip 601.
In order to further improve the uniformity of solution mixing, a lifting seat 7 capable of performing reciprocating linear motion in the front-back direction and lifting is connected in the incubator 1, a plurality of micro air inlet devices 14 which are in one-to-one correspondence with chips 601 in any group of chip assemblies 6 are arranged on the lower side of the lifting seat 7, a main air pipe 11 is fixedly connected below the lifting seat 7, an air inlet valve 13 is connected to the output end of the air inlet device 14, an air distribution pipe 1102 is connected to the output end of the air inlet valve 13, a composite air hole a is formed in the upper end of the chip 601, an inserting pipe 1102a corresponding to the composite air hole a is fixed on the lower side of the tail end of the air distribution pipe 1102, one side, away from the inserting pipe 1102a, of the air distribution pipe 1102 is connected to the main air pipe 11, an air suction one-way valve 12 is connected to the air distribution pipe 1102 between the inserting pipe 1102a and the main air pipe 11, a side end of the main air pipe 11 is connected with a negative pressure air pipe 1101, a plurality of air suction holes b which are in one-to-one correspondence with the chip assemblies 6 are arranged on one end of the upper side of the culture plate 21, a negative pressure air suction pipe 1101a is fixedly arranged on the negative pressure air suction pipe 1101a corresponding to the air suction holes 1101a, and the negative pressure air suction pipe corresponding to the air suction holes a can be inserted into the air suction holes 1101a right through the inserting pipe 21 b; the incubator 1 under the support plate 102 is fixedly connected with a pipeline connection driver, the pipeline connection driver is connected with a telescopic rod 16 capable of performing reciprocating linear motion in the height direction, the upper end of the telescopic rod 16 is connected with a joint 18, the joint 18 is connected with a negative pressure connecting pipe 17, the negative pressure connecting pipe 17 can be just inserted into the culture plate 21 through a negative pressure connecting hole c, and one end of the negative pressure connecting pipe 17 away from the joint 18 is connected with the negative pressure equipment 5.
In the initial state, the negative pressure connection pipe 17 is below the culture plate 21; when the solution needs to be promoted to flow in the direction of the reaction tank, the pipeline connection driver acts to enable the telescopic rod 16 to move upwards, the telescopic rod 16 drives the negative pressure connecting pipe 17 to move upwards through the connector 18, and when the negative pressure connecting pipe 17 is inserted into the culture plate 21 through the negative pressure connecting hole c, the pipeline connection driver stops acting; the lifting seat 7 moves to the position of the corresponding chip assembly 6, the lifting seat 7 descends, the negative pressure cannula 1101a is inserted into the culture plate 21 through the air suction hole b, the negative pressure equipment 5 is opened, the corresponding air suction check valve 12 is controlled to open and close according to the conditions of each chip 601, the flow of the solution in the corresponding chip 601 in the mixing flow channel of the chip 601 is promoted, the air suction is finished, the air inlet pump and the air inlet valve 13 above the corresponding chip 601 are controlled to open, the air is fed into the chip 601, the solution reversely flows in the mixing flow channel of the chip 601, the solution continuously flows back and forth, and the uniformity of the solution mixing is improved.
In order to further realize the lifting of the lifting seat 7, two lower adjusting assemblies 10 for adjusting the position of the lifting seat 7 are further connected in the incubator 1, the two lower adjusting assemblies 10 are respectively arranged on the left side and the right side of the lifting seat 7, each lower adjusting assembly 10 comprises a transverse frame 1007 fixedly connected to the inner side of the incubator 1, a lower transmission screw 1006 which is horizontally arranged is rotatably connected to the transverse frame 1007, a first sliding table 1008 which is in sliding connection with the transverse frame 1007 is connected to the lower transmission screw 1006 in a threaded manner, a vertical stand 1002 is fixedly connected to one side of the first sliding table 1008 opposite to the lifting seat 7, a lifting screw 1003 which is vertically arranged is rotatably connected to the vertical stand 1002, a second sliding table 1004 which is in sliding connection with the vertical stand 1002 is connected to the lifting screw 1003 in a threaded manner, a lifting motor 1001 is fixedly connected to the upper side of the vertical stand 1002, and the lifting motor 1001 is connected to the lifting screw 1003, one end of the transverse frame 1007 in the front-rear direction is fixedly connected to the lower transmission motor 1005 and the lower transmission screw 1006 is connected to the lower transmission screw 1005.
When the positions of the negative pressure insertion pipe 1101a and the insertion pipe 1102a are adjusted, the positions of the lifting seat 7 in the front-rear direction are adjusted firstly, specifically, the lower transmission motor 1005 acts, the lower transmission screw 1006 rotates, the lower transmission screw 1006 drives the vertical frame 1002 to move in the front-rear direction through the first sliding table 1008, and when the negative pressure insertion pipe 1101a is aligned with the corresponding negative pressure insertion hole and the insertion pipe 1102a is aligned with the corresponding composite air hole a, the lower transmission motor 1005 stops acting; the lifting motor 1001 is operated, the lifting screw 1003 is rotated, the second sliding table 1004 is moved, the movement direction of the lifting motor 1001 is controlled, the lifting seat 7 is lowered, when the negative pressure cannula 1101a and the inserting tube 1102a are respectively inserted into the culture plate 21 and the corresponding chip 601, the lifting motor 1001 is stopped, the solution mixing is finished, the lifting motor 1001 is operated reversely, the lifting seat 7 is reset, the lifting motor 1001 is stopped, the lower transmission motor 1005 is operated, the lifting seat 7 and the injector 8 are dislocated to the proper positions, and the lower transmission motor 1005 is stopped.
In order to further realize the position adjustment of the injector 8 in the front-rear direction, an upper adjusting component 15 for adjusting the position of the movable seat 27 is connected in the incubator 1 above the supporting plate 102, the upper adjusting component 15 comprises a fixed plate 101 arranged in front of the movable seat 27 and fixed on the inner side of the incubator 1, an upper transmission screw 1501 is rotatably connected between the fixed plate 101 and the incubator 1, the movable seat 27 is in threaded connection with the upper transmission screw 1501, guide rods 1503 are respectively connected to the fixed plates 101 on the left side and the right side of the upper transmission screw 1501, the guide rods 1503 penetrate through the movable seat 27 and then are connected to the incubator 1, an upper transmission motor 1502 is fixedly connected to the front side of the fixed plate 101, and the upper transmission motor 1502 is connected with the upper transmission screw 1501.
When the position of the syringe 8 is adjusted, the upper transmission motor 1502 is operated, the upper transmission screw 1501 is rotated, the upper transmission screw 1501 drives the movable base 27 to move, and when the syringe 8 is moved to a desired position, the upper transmission motor 1502 stops operating.
Specifically, the incubator further comprises a rear accommodating box 32 arranged at the rear side of the incubator 1, the rear accommodating box 32 is connected with a liftable storage rack 22, the lifting structure of the storage rack 22 is a structure in the prior art, preferably a scissor type lifting structure, and as can be seen from fig. 7, the specific structure is not repeated here, and is not an improvement point of the invention; the rack 22 is provided with a plurality of storage plates 2201 for storing the culture plates 21 in a height direction, a rear storage box 32 behind the culture plates 21 is connected with a push-pull assembly capable of pushing the culture plates 21 to be arranged on a supporting plate 102 or pulling the culture plates 21 to be arranged on the storage plates 2201, the rear side of the culture plates 21 is fixedly connected with steel bars 26, the push-pull assembly comprises at least one linear driver 24 fixedly connected in the rear storage box 32 and behind the rack 22, the linear driver 24 is preferably an electric push rod, the linear driver 24 is connected with a push-pull rod 23 capable of conducting reciprocating linear movement in the front-rear direction, the tail end of the push-pull rod 23 is connected with an electromagnet 25, when the electromagnet 25 is powered on, the steel bars 26 are adsorbed on the electromagnet 25, and when the electromagnet 25 is powered off, the steel bars 26 are separated from the electromagnet 25.
Through the joint setting of liftable supporter 22 and push-and-pull subassembly, conveniently realize the chip 601 of different culture plates 21 and cultivate, improve chip 601 and cultivate efficiency.
According to the invention, through the joint arrangement of the components such as the multichannel peristaltic pump 3, the liquid storage shell 2, the injector 8 and the like, the sample adding of each chip 601 in a group of chip assemblies 6 is realized; through the joint arrangement of the main air pipe 11, the air distribution pipe 1102, the air suction check valve 12, the air inlet pump and the air inlet valve 13, independent circulation mixing of solutions in different chips 601 is realized, and the uniformity of solution mixing in the culture process is improved, so that the later culture effect is improved.
Example 2
Referring to fig. 13 and 14, a second embodiment of the present invention provides a chip culturing apparatus which is different from the first embodiment 1 in that it enables positional adjustment of the syringe 8.
Specifically, the lower side of the moving seat 27 is connected with a position adjusting seat 29, the lower side of the moving seat 27 is provided with two sliding rails 31 which are arranged at intervals in the front-rear direction, the position adjusting seat 29 is slidably connected to the sliding rails 31, a plurality of lifting drivers 9 are connected to the lower side of the position adjusting seat 29, a position adjusting motor 30 is fixedly connected to the forward side of the moving seat 27, the left and right ends in the moving seat 27 are respectively and rotatably connected with a driving wheel and a driven wheel (this is not shown in the prior art), the driving wheel is connected with the driven wheel through a driving belt 28, and the position adjusting seat 29 is fixedly connected to the lower side of the driving belt 28 below; the upward end of the chip 601 is also provided with a waste liquid port e, and a waste liquid pool mutually independent of the reaction pool is arranged in the chip 601 at the lower side of the waste liquid port e.
When the liquid outlet branch pipe 1901, the liquid outlet main pipe 19 and the corresponding channels and pipelines of the multi-channel peristaltic pump 3 are required to be cleaned, the position adjusting motor 30 acts to align the injector 8 with the corresponding liquid outlet e, the injector 8 descends and is inserted into the liquid outlet e, the liquid outlet one-way valve 20 corresponding to the multi-channel peristaltic pump 3 and the liquid storage tank 201a filled with cleaning liquid is opened, the cleaning liquid sequentially flows through the liquid outlet branch pipe 1901, the liquid outlet total and the corresponding channels and pipelines of the multi-channel peristaltic pump 3, so that the corresponding channels and pipelines are cleaned, new reaction solution is injected subsequently, the cleaning is finished, and the multi-channel peristaltic pump 3 and the corresponding liquid outlet one-way valve 20 are closed.
Example 3
Referring to fig. 3 and 4, for a third embodiment of the present invention, which is based on the second embodiment, the present embodiment provides a method for culturing a chip 601 using a chip culturing apparatus, comprising the steps of,
lifting the culture plate 21 with a plurality of chips 601 to be cultured to the height of the support plate 102, enabling the linear driver 24 to act, enabling the push-pull rod 23 to extend forwards, enabling the electromagnet 25 to be attached to the steel bar 26, enabling the electromagnet 25 to be powered on, and stopping the linear driver 24 when the culture plate 21 is pushed to a proper position on the support plate 102;
the position of the movable seat 27 is regulated, so that the injector 8 is moved to the position of the forefront chip assembly 6, the position of the injector 8 in the left-right direction is regulated, the injector 8 to be injected is sequentially aligned with the liquid inlet f of the corresponding chip 601, the corresponding lifting driver 9 acts when injecting liquid, the injector 8 is lowered, when the injection needle of the injector 8 is inserted into the chip 601 through the corresponding liquid inlet f, the lifting driver 9 stops acting, the corresponding liquid outlet check valve 20 is controlled to be opened according to the set liquid injection sequence, the multichannel peristaltic pump 3 works, so that the solution in the corresponding channel is pumped into the injector 8, and when the liquid inlet reaches the set liquid inlet threshold, the corresponding liquid outlet check valve 20 and the multichannel peristaltic pump 3 are closed, the lifting driver 9 acts reversely, resets, and the lifting driver 9 stops acting;
when the solution in the chip 601 is required to be further mixed after one-stage liquid injection is finished, the movable seat 27 is moved backwards, the lower transmission motor 1005 acts, the lifting seat 7 is moved forwards to a set position, the lifting seat 7 descends, the plug tube 1102a is inserted into the chip 601 through the corresponding composite air hole a, the negative pressure equipment 5 works, the corresponding air suction one-way valve 12 is controlled to be opened, the solution in the liquid storage tank of the chip 601 flows to the direction of the reaction tank through the mixed flow channel in the chip 601, when the set first time threshold is reached, the negative pressure equipment 5 and the air suction one-way valve 12 are closed, the air inlet pump is opened, the solution flows to the direction away from the reaction tank, and when the set second time threshold is reached, the air inlet pump is closed; repeating the above actions, when the mixing time reaches a set third time threshold, opening the negative pressure equipment 5 to enable the solution to enter the reaction tank, closing the negative pressure equipment 5, moving the lifting seat 7 backwards, moving the moving seat 27 forwards to perform the liquid injection work of the next stage, and otherwise, returning to the mixing step;
after the first injection of one row of chip assemblies 6 is finished, the movable seat 27 is moved to the position of the next row of chip assemblies 6, the steps are repeated until a plurality of groups of chip assemblies 6 in the culture plate 21 are all subjected to the first injection, the injector 8 is lifted and reset, the position of the injector 8 is adjusted, the injector 8 is aligned to the waste liquid port e, the injector 8 is lowered to be inserted into the waste liquid port e, the liquid outlet one-way valve 20 of the liquid storage tank 201a filled with cleaning liquid is controlled to be opened, the multi-channel peristaltic pump 3 is opened, the cleaning liquid is pumped into the waste liquid pool, all pipelines are cleaned at the position of the last row of chip assemblies 6, the cleaning is finished, the first injection is finished, and the second injection is started from the last row of chip assemblies 6;
the second liquid injection process is similar to the first liquid injection process, all groups of chip assemblies 6 complete the second liquid injection, the cleaning of each pipeline is completed at the corresponding position of the forefront chip assembly 6, and the second liquid injection process is finished;
repeating the above actions until all liquid injection is completed;
the cell culture environment of the incubator 1 (this is the prior art) is set, the temperature in the incubator 1 is controlled at a set temperature, the carbon dioxide content is controlled within a set range value, the cell culture is performed, the cell culture is finished, the toxicity test of the cell can be performed, the toxicity reaction is finished, the linear driver 24 acts reversely, the push-pull rod 23 is retracted backwards, the culture plate 21 is pulled to the corresponding placing plate 2201, the electromagnet 25 is powered off, and the culture plate 21 is released.
It should be noted that the solutions in the liquid storage portions 201 corresponding to different chip assemblies 6 may be different to achieve different cell cultures.
Taking cultured liver cells as an example, the use method of the liver cells is further described, and the method comprises the following steps:
opening the upper cover of the liquid storage shell 2, wherein each five continuous liquid storage tanks 201a are in a group, wherein a first liquid storage tank 201a in each group is added with a mixed solution of human liver primary cells, human immune cells and human astrocytes, a second liquid storage tank 201a is added with a solution of human endothelial cells, a third liquid storage tank 201a is added with a nutrient solution required by cells, a fourth liquid storage tank 201a is added with a cleaning solution, and a fifth liquid storage tank 201a is added with a certain test drug;
lifting the culture plate 21 with a plurality of chips 601 to be cultured to the height of the support plate 102, enabling the linear driver 24 to act, enabling the push-pull rod 23 to extend forwards, enabling the electromagnet 25 to be attached to the steel bar 26, enabling the electromagnet 25 to be powered on, and stopping the linear driver 24 when the culture plate 21 is pushed to a proper position on the support plate 102;
the position of the movable seat 27 is adjusted through the upper transmission motor 1502 and the screw rod, so that the injector 8 is moved to the position of the forefront chip assembly 6, the position of the injector 8 in the left-right direction is adjusted through the synchronous belt, the injector 8 to be injected is sequentially aligned with the liquid inlet f of the corresponding chip 601, when the liquid is injected, the corresponding lifting driver 9 acts, the injector 8 descends, when the injection needle of the injector 8 is inserted into the chip 601 through the corresponding liquid inlet f, the lifting driver 9 stops acting, at the moment, the liquid outlet one-way valve 20 corresponding to the first liquid storage tank 201a is opened, the mixed solution of human liver primary cells, human immune cells and human astrocytes flows out of the liquid outlet header 19, the multichannel peristaltic pump 3 works, the corresponding liquid outlet one-way valve 20 and the multichannel peristaltic pump 3 are closed when the set first liquid inlet time threshold is reached, the lifting driver 9 reversely acts, and resets, and the lifting driver 9 stops acting;
when the first injection is finished and three cell solutions in the chip 601 need to be mixed, the movable seat 27 is moved backwards, the lower transmission motor 1005 acts, the lifting seat 7 is moved forwards to a set position, the lifting seat 7 descends, the plug tube 1102a is inserted into the chip 601 through the corresponding composite air hole a, the negative pressure equipment 5 works, the corresponding air suction one-way valve 12 is controlled to be opened, the solution in the liquid storage tank of the chip 601 flows towards the direction of the reaction tank through the mixed flow channel in the chip 601, when the set first time threshold is reached, the negative pressure equipment 5 and the air suction one-way valve 12 are closed, the air inlet pump is opened, the solution flows away from the direction of the reaction tank, when the set second time threshold is reached, the air inlet pump is closed, the above acts are repeated, when the mixing time reaches the set third time threshold, the mixing is finished, the negative pressure equipment 5 is opened, the solution enters the reaction tank, the negative pressure equipment 5 is closed, the lifting seat 7 moves backwards, the movable seat 27 moves forwards, and the next injection work is carried out, otherwise, the mixing step is returned;
after the first liquid injection of one row of chip assemblies 6 is finished and the mixing is finished, the positions of the movable seats 27 are regulated through the upper transmission motor 1502 and the screw rods, the positions of the next row of chip 601 groups are reached, the process is repeated, so that after the first liquid injection of the chip assemblies 6 of the following rows is finished, all groups of chip assemblies 6 are controlled to be opened by the liquid outlet one-way valves 20 corresponding to the fourth liquid storage groove 201a after the first liquid injection is finished, the multi-channel peristaltic pump 3 is opened, the cleaning liquid is pumped into the waste liquid tank, all pipelines are cleaned, the cleaning is finished, the first liquid injection is finished, and the second liquid injection is started from the last row of chip assemblies 6;
the second liquid injection process is similar to the first liquid injection process, and the process is that the liquid outlet one-way valve 20 of the second liquid storage tank 201a works to add cell nutrient solution into the chip 601, so that liver cells obtain basic conditions for culture, and cleaning of each pipeline is completed at the corresponding position of the forefront chip assembly 6;
the third liquid injection process is similar to the first liquid injection process, and liquid injection is started from the forefront chip assembly 6, wherein the process is that a liquid outlet one-way valve 20 of a third liquid storage groove 201a works, and cell nutrient solution is added into a chip 601, so that liver cells obtain basic conditions for culture, and cleaning of each pipeline is completed at the corresponding position of the final chip assembly 6;
after the injection of all culture plates 21 is finished, the heater in the incubator 1 starts to work to raise the temperature, a plurality of fans are arranged in the incubator 1, the fans send heat in the incubator 1 to all parts in the incubator 1, the heat distribution is more uniform, the temperature in the incubator 1 is kept at a proper temperature through a temperature control module, proper conditions (which are the prior art and related parts are not specifically marked) are provided for cell culture, the culture of liver cells can be carried out, and certain drugs in the fifth liquid storage tank 201a can be used for toxicity test of future drugs on the liver cells, namely, the test drugs are injected into the cultured chips 601 through the injector 8 to react for a period of time;
after the reaction is finished, the linear driver 24 acts reversely, so that the push-pull rod 23 retracts backwards, the culture plate 21 is pulled to the corresponding storage plate 2201, the electromagnet 25 is powered off, the culture plate 21 is loosened, the lifting table is lifted, the height of the next culture plate 21 is consistent with that of the support plate 102, the linear driver 24 acts, the push-pull rod 23 extends forwards, the electromagnet 25 is attached to the steel bar 26, the electromagnet 25 is powered on, the culture plate 21 is pushed to the proper position on the support plate 102, and the chip 601 liquid injection culture process is repeated.
When toxicity detection is needed for the medicine, the corresponding culture plate 21 can be pushed onto the supporting plate 102 in the incubator 1, the corresponding culture plate 21 is taken out, cell dye is injected into the chip 601, a professional instrument is taken to shoot a fluorescent chart, and the imaging J software is used for statistical analysis, so that the number of surviving cells and the number of dying cells can be known, and if the concentration of the medicine is positively correlated with the death rate of the cells, the medicine has hepatotoxicity.
It should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered in the scope of the claims of the present invention.

Claims (10)

1. A chip culture device, characterized in that: including having the incubator that holds the chamber, incubator internal fixation has the backup pad, swing joint has the culture plate in the backup pad, a plurality of groups of chip assemblies that wait to cultivate have been arranged in the front and back direction of culture plate, the chip assemblies comprises a plurality of chips that set up in the left and right directions upper interval, incubator internal connection above the culture plate has the movable seat that can do reciprocal rectilinear motion, the lower extreme of movable seat has arranged a plurality of syringes that can go up and down, incubator outside fixedly connected with multichannel peristaltic pump and stock solution casing, have a plurality of groups of mutually independent stock solution portions in the stock solution casing, the stock solution portion includes a plurality of mutually independent stock solution groove, a plurality of passageway inputs of multichannel peristaltic pump respectively with a plurality of groups stock solution portion one-to-one, a plurality of passageway outputs of multichannel peristaltic pump respectively with a plurality of syringes one-to-one, the solution in the stock solution portion can be pumped into the syringe in proper order.
2. The chip culture apparatus of claim 1, wherein: the liquid outlet main pipe is connected with a plurality of liquid outlet branch pipes which are in one-to-one correspondence with liquid storage tanks in the same group of liquid storage parts, a liquid outlet one-way valve is connected on the liquid outlet branch pipes, and the liquid outlet main pipe is connected with corresponding channel input ends.
3. The chip culture apparatus of claim 2, wherein: the upper portion of chip is opened there is the compound gas pocket, the tip of culture plate has the air cavity, and a plurality of ability and air cavity intercommunication and with the bleed hole of chip subassembly one-to-one have been arranged to the upper end of culture plate, the lower extreme of culture plate is opened there is the negative pressure connecting hole, and incubator outside fixedly connected with negative pressure equipment, and negative pressure equipment can be connected with the culture plate through the negative pressure connecting hole, the incubator internal connection has can be in the back-and-forth direction reciprocating rectilinear motion and can go up and down the lifting seat, the lower side of lifting seat has arranged a plurality of and arbitrary chip in the chip subassembly one-to-one's of group inlet equipment, lifting seat below fixedly connected with total trachea, inlet valve is connected with to inlet valve's output, and the output of inlet valve is connected with the gas distribution pipe, and the downside at the gas distribution pipe end is fixed with the spliced tube that corresponds with compound gas pocket, and the gas distribution pipe is kept away from one side of spliced tube is connected with on the total trachea, is connected with the check valve that bleeds on the gas distribution pipe between spliced tube and the total trachea.
4. The chip culture apparatus of claim 3, wherein: the side end of the main air pipe is connected with a negative pressure air pipe, a negative pressure intubation tube which is vertically arranged and corresponds to the air suction hole is fixed on the negative pressure air pipe, and the negative pressure intubation tube can be just inserted into the culture plate through the corresponding air suction hole.
5. The chip culture apparatus of claim 3 or 4, wherein: the incubator of backup pad below internal fixedly connected with pipe connection driver, be connected with the telescopic link that can reciprocate rectilinear motion in the direction of height on the pipe connection driver, the upper end of telescopic link is connected with the joint, be connected with the negative pressure connecting pipe on the joint, the negative pressure connecting pipe can just be inserted in the culture plate through the negative pressure connecting hole, and the negative pressure connecting pipe is kept away from the one end and the negative pressure equipment connection of joint.
6. The chip culture apparatus of claim 3 or 4, wherein: the incubator is characterized in that a lower adjusting component used for adjusting the position of the lifting seat is further connected in the incubator, the lower adjusting component comprises a transverse frame fixedly connected to the inner side of the incubator, a lower transmission screw rod which is horizontally arranged is rotatably connected to the transverse frame, a first sliding table which is connected to the transverse frame in a sliding mode is connected to the lower transmission screw rod in a threaded mode, a vertical stand is fixedly connected to one side of the first sliding table, which is vertically arranged, of the lifting seat, a second sliding table which is connected to the vertical stand in a sliding mode is rotatably connected to the lifting screw rod, the lifting seat is fixedly connected to the second sliding table, a lifting motor is fixedly connected to the upper side of the vertical stand, the lifting motor is connected to the lifting screw rod, a lower transmission motor is fixedly connected to one end of the transverse frame in the front-rear direction, and the lower transmission motor is connected to the lower transmission screw rod.
7. The chip culture apparatus of claim 6, wherein: the incubator of backup pad top is connected with the last adjusting part that is used for adjusting and removes the seat position, go up adjusting part including setting up in the place ahead of removing the seat and fixing the fixed plate inboard at the incubator, rotationally be connected with the transmission lead screw between fixed plate and the incubator, it is last on the transmission lead screw to remove seat threaded connection, be connected with the guide arm on the fixed plate of transmission lead screw left and right sides respectively, the guide arm passes to remove the seat and connects on the incubator, fixed plate front side fixedly connected with goes up the transmission motor, go up the transmission motor and go up the transmission lead screw connection.
8. The chip culture apparatus of claim 7, wherein: the device is characterized by further comprising a rear containing box arranged at the rear side of the incubator, wherein a storage rack capable of lifting is connected in the rear containing box, a plurality of storage plates used for storing the culture plates are arranged in the height direction of the storage rack, and a push-pull assembly capable of pushing the culture plates to be placed on the support plates or pulling the culture plates to be placed on the storage plates is connected in the rear containing box of the culture plates.
9. The chip culture apparatus of claim 8, wherein: the rear side fixedly connected with billet of culture plate, push-and-pull subassembly includes at least one fixed connection at the rearmounted straight line driver that holds the incasement and be at the supporter rear, be connected with the push-and-pull rod that can do reciprocal rectilinear movement in the fore-and-aft direction on the straight line driver, the end-to-end connection of push-and-pull rod has the electro-magnet, and when the electro-magnet was on, the billet was adsorbed on the electro-magnet, and the electro-magnet breaks away from the electro-magnet when the electro-magnet was cut off the power supply.
10. A method of culturing using the chip culturing apparatus of claim 9, characterized in that: comprises the steps of,
lifting a culture plate provided with a plurality of chips to be cultured to the height of a supporting plate, enabling a linear driver to act, enabling a push-pull rod to extend forwards, enabling an electromagnet to be attached to a steel bar, enabling the electromagnet to be electrified, and stopping the linear driver when the culture plate is pushed to a proper position on the supporting plate;
the position of the movable seat is regulated to enable the injector to be moved to the position of the forefront chip assembly, the position of the injector in the left-right direction is regulated to enable the injector to be injected to be aligned with the liquid inlet of the corresponding chip in sequence, the corresponding lifting driver acts to enable the injector to descend, when the injection needle of the injector is inserted into the chip through the corresponding liquid inlet, the lifting driver stops acting, the corresponding liquid outlet one-way valve is controlled to be opened according to the set liquid injection sequence, the multichannel peristaltic pump works to enable the solution in the corresponding channel to be pumped into the injector, and when the liquid inlet amount reaches the set liquid inlet amount threshold, the corresponding liquid outlet one-way valve and the multichannel peristaltic pump are closed, the lifting driver acts reversely to reset, and the lifting driver stops acting;
when the solution in the chip is required to be mixed after the injection at one stage is finished, the movable seat is moved backwards, the lower transmission motor acts, the lifting seat is moved forwards to a set position, the lifting seat is lowered, the plug-in pipe is inserted into the chip through the corresponding composite air hole, the negative pressure equipment works, the corresponding air suction check valve is controlled to be opened, the solution in the liquid storage tank of the chip flows to the direction of the reaction tank through the mixing flow passage in the chip, when the set first time threshold value is reached, the negative pressure equipment and the air suction check valve are closed, the air inlet pump is opened, the solution flows in the direction away from the reaction tank, when the set second time threshold value is reached, the air inlet pump is closed, when the mixing time reaches the set third time threshold value, the negative pressure equipment is opened, the solution enters the reaction tank, the negative pressure equipment is closed, the lifting seat is moved backwards, the movable seat moves forwards, the injection at the next stage is performed, and otherwise, the mixing step is returned;
after the liquid injection culture of one row of chip assemblies is finished, the movable seat moves back to the position of the next batch of chip assemblies, the steps are repeated until a plurality of groups of chip assemblies in the culture plate are subjected to the chip culture, and the next step is carried out;
the linear driver acts reversely to enable the push-pull rod to retract backwards, the culture plate is pulled to the corresponding storage plate, the electromagnet is powered off, and the culture plate is loosened.
CN202310555267.6A 2023-05-17 2023-05-17 Chip culture device and application method thereof Active CN116536155B (en)

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