CN113234583A - Cell reaction module integrated with gas circuit control and method for carrying out cell reaction by adopting cell reaction module - Google Patents
Cell reaction module integrated with gas circuit control and method for carrying out cell reaction by adopting cell reaction module Download PDFInfo
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- CN113234583A CN113234583A CN202110534947.0A CN202110534947A CN113234583A CN 113234583 A CN113234583 A CN 113234583A CN 202110534947 A CN202110534947 A CN 202110534947A CN 113234583 A CN113234583 A CN 113234583A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- 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
Abstract
The invention provides a cell reaction module controlled by an integrated gas circuit and a method for carrying out cell reaction by adopting the cell reaction module, wherein the cell reaction module comprises an integrated gas circuit control plate and a cell reaction plate which are mutually attached, at least two mutually independent driving gas circuit channels are arranged in the integrated gas circuit control plate, reagent grooves with the same number as the driving gas circuit channels are arranged on the surface of one side of the cell reaction plate attached to the integrated gas circuit control plate, and each driving gas circuit channel is independently communicated with one reagent groove; the cell reaction plate is provided with a reaction cavity on the surface of one side far away from the integrated gas circuit control plate, and the reaction cavity is communicated with the reagent tank. The invention integrates all the driving gas path channels into a whole by slotting in the integrated gas path control plate, so that the flow path control process of the reaction reagent is stable and reliable, and the whole cell reaction module is convenient to install and occupies small space.
Description
Technical Field
The invention belongs to the technical field of cell reaction, and relates to a cell reaction module integrated with gas circuit control and a method for carrying out cell reaction by adopting the cell reaction module.
Background
Cells are the fundamental unit of biology and researchers are making more effort to attempt to isolate, study and compare them individually. Single cell sequencing refers to the relatively simple genome of single cell microorganisms, larger and more complex human cell genomes that have been sequenced in DNA research.
The single cell sequencing mainly comprises single cell genome sequencing and transcriptome sequencing, and the change conditions of the genome and the transcriptome of a single cell are respectively revealed through sequence analysis of DNA and RNA in the single cell. Single cell whole genome sequencing is non-selective and homogeneous amplification of the whole genome sequence of selected target cell, and subsequent high-throughput sequencing with exon trapping technology. The sequencing of the single cell transcriptome is to utilize a high-throughput sequencing technology to perform cDNA sequencing so as to obtain almost all transcripts of a specific organ or tissue in a certain state, is mainly used for excavating a gene regulation network in the whole genome range, and is particularly suitable for stem cells with high heterogeneity and cell populations at the early development stage of embryos. In combination with living cell imaging systems, single cell transcriptome analysis is more helpful for the in-depth understanding of cell differentiation, cell reprogramming and transdifferentiation processes and related gene regulatory networks.
CN208104383U discloses a microfluidic chip for efficient single-cell droplet preparation, which comprises: the single cell culture device comprises a reaction solution inlet, a marker solution inlet and a single cell inlet, wherein the reaction solution inlet is communicated with a cell liquid storage container, and a stirring device is arranged in the cell liquid storage container; an oil phase inlet; the inlet of the single cell channel is communicated with the single cell inlet; the inlet of the liquid mixing channel is respectively communicated with the outlet of the single cell channel, the reaction liquid inlet and the marker solution inlet; the inlet of the liquid drop generating channel is communicated with the outlet of the liquid mixing channel and the oil phase inlet, and the oil phase is wrapped on the surface of the single cell in the liquid drop generating channel to form single cell liquid drops; and a droplet generation outlet in communication with the outlet of the droplet generation channel.
CN103571738A discloses a micro-fluidic chip device based on chemokine enrichment effect and a preparation method thereof, which comprises a PDMS substrate layer, two corresponding micro-fluidic chip modules A and B, a semi-permeable membrane arranged between the two micro-fluidic chip modules, a top cover plate and corresponding sample inlet and outlet pipelines. The micro-fluidic chip module A is used for injecting and enriching and sorting cell samples to be processed, and a sample pool in the module A is connected with a sample inlet 2 and a sample outlet 2, so that the injection of the cell samples to be processed and the sorting of tissue stem cells enriched on a semipermeable membrane are realized; the module B is used for injecting chemotactic factors, and chemotactic effect is formed in the area, close to the semipermeable membrane, of the sample pool of the module A by virtue of the semipermeable membrane. The stem cells of various tissues in the cell sample injected into the module A move towards the semipermeable membrane under the chemotactic effect and are separated from other cells, and the separated stem cells and other cell samples are collected through different sample outlets.
CN108117968A discloses a method for high-throughput automatic single cell capture based on droplet microfluidic chip. The microfluidic chip comprises two layers, wherein the upper layer is a flow path inlet and outlet layer; the lower layer is a flow path control layer; the flow path inlet and outlet layer has a liquid flow path channel inlet and a liquid flow path channel outlet; the flow path control layer consists of a single cell capturing flow path channel, a gas path channel and a liquid drop generating unit. The method introduces a gas flow channel with controllable air pressure, can form a negative pressure flow channel and automatically suck the single cell suspension into the capture trap, and is convenient for observing and detecting the proliferation, differentiation, drug reaction and other behaviors of the single cell.
There is the part that relates to the gas circuit system among the unicellular sample preparation chip on the market at present, and the most is connected components such as various switching valves of syringe intercommunication and pressure sensor, but each part need be through connecting the conversion, gathers at the conversion process, produces unstable deviation easily, and how to inherit this gas circuit system as an organic whole, guarantees the stable performance when reducing the part volume, is the technical problem that awaits the solution at present.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide an integrated gas circuit controlled cell reaction module and a method for carrying out cell reaction by adopting the integrated gas circuit controlled cell reaction module. In the use, inject into drive gas in the drive gas way passageway of difference, can inject the reaction reagent of different reagent inslots into the reaction chamber one by one, the injection process of different reaction reagent is independent each other and does not influence for the flow path control process of reaction reagent is reliable and stable, whole cell reaction module simple to operate and occupation space are little.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides an integrated gas circuit controlled cell reaction module, which comprises an integrated gas circuit control board and a cell reaction board which are mutually attached, wherein at least two mutually independent driving gas circuit channels are arranged in the integrated gas circuit control board, the surface of one side of the cell reaction board, which is attached to the integrated gas circuit control board, is provided with reagent grooves with the same number as the driving gas circuit channels, and each driving gas circuit channel is independently communicated with one reagent groove; the cell reaction plate is characterized in that a reaction cavity is formed in the surface of one side, away from the integrated gas circuit control plate, of the cell reaction plate, the reaction cavity is communicated with a reagent groove, a reaction reagent is injected into the reagent groove in advance, a driving gas is injected into the reagent groove through a driving gas circuit channel, and the reaction reagent in the reagent groove flows into the reaction cavity under the driving of the pressure of the driving gas.
According to the invention, all the driving gas channels are integrated into a whole in a manner of slotting in the integrated gas circuit control board, and part of the driving gas channels are communicated through the electromagnetic valve, so that the communication or independence among different driving gas channels is realized according to requirements through controlling a switch by an upper computer, and the driving gas is used as the driving force for injecting the reaction reagent to carry out flow path conveying of the reaction reagent. In the use, inject into drive gas in the drive gas way passageway of difference, can inject the reaction reagent of different reagent inslots into the reaction chamber one by one, the injection process of different reaction reagent is independent each other and does not influence for the flow path control process of reaction reagent is reliable and stable, whole cell reaction module simple to operate and occupation space are little.
It should be noted that the opening mode of the driving gas channel is not specifically required or limited, and the smooth transmission of the driving gas can be satisfied. Illustratively, the following two schemes may be optionally employed:
the first scheme is as follows: the integrated gas circuit control board is of an integrated structure, and the driving gas circuit channel is directly arranged in the integrated gas circuit control board in a pouring, drilling or additive manufacturing mode and the like;
scheme II: the integrated gas circuit control board is of a split structure and is formed by laminating and attaching a gas circuit upper board and a gas circuit lower board, a gas circuit groove is formed in an attaching surface between the gas circuit upper board and/or the gas circuit lower board, and after the gas circuit upper board and the gas circuit lower board are attached, the gas circuit groove is sealed to form a driving gas circuit channel; it can be understood that, in this embodiment, the air channel groove may be opened on the lower surface of the air channel upper plate or the upper surface of the air channel lower plate, or may be opened on both the lower surface of the air channel upper plate and the upper surface of the air channel lower plate.
It should be noted that, the cell reaction plate is also provided with a micro channel for flowing the reaction reagent, the manner of providing the micro channel can be referred to the above description about the driving gas channel, that is, the cell reaction plate can be an integrated structure or a split structure, when the cell reaction plate is an integrated structure, the micro channel is directly provided inside the cell reaction plate by casting, drilling or additive manufacturing; certainly, the cell reaction plate can be a split structure formed by laminating and attaching the reaction upper plate and the reaction lower plate, the attachment surface of the reaction upper plate and/or the reaction lower plate is provided with a flow channel groove, and the flow channel groove is sealed to form the microchannel after the reaction upper plate and the reaction lower plate are attached.
It should be noted that, the number of reagent grooves is not specifically required, the number of reagent grooves is the same as that of the driving gas channels, and the gas outlet end of each driving gas channel corresponds to one reagent groove, so that those skilled in the art can design the number and positions of reagent grooves in a targeted manner according to different cell reactions.
As a preferred technical scheme of the invention, the cell reaction plate and the integrated gas circuit control plate are jointed on one side surface thereof with a buffer groove, the reagent groove and the reaction cavity are respectively and independently communicated with the buffer groove, the driving gas circuit channel, the reagent groove and the buffer groove are sequentially communicated along the flowing direction of the driving gas, the driving gas is introduced into the reagent groove through the driving gas circuit channel, and the reaction reagents stored in the reagent grooves enter the buffer groove one by one and are injected into the reaction cavity through the buffer groove.
Preferably, an electromagnetic valve is arranged at the air inlet end of the driving air channel and used for controlling the introduction amount of the driving air.
In the invention, the electromagnetic valve connected to the driving gas channel is mainly used for controlling the flow of the driving gas, when the cell reaction module provided by the invention is used, the reaction reagent stored in the reagent tank is pressed into the buffer tank under the driving of the pressure of the driving gas, and the flow of the driving gas is regulated through the electromagnetic valve, so that the injection amount of the reaction reagent entering the buffer tank is changed.
As a preferred technical scheme of the present invention, a waste liquid tank is further disposed on a side surface of the cell reaction plate, which is attached to the integrated gas circuit control plate, the waste liquid tank is communicated with the reaction chamber, a waste liquid pumping channel is disposed inside the integrated gas circuit control plate, the waste liquid pumping channel is communicated with the waste liquid tank, the reaction chamber, the waste liquid tank and the waste liquid pumping channel are sequentially communicated along a pumping direction, and the waste liquid after the reaction in the reaction chamber is pumped into the waste liquid tank by pumping air through the waste liquid pumping channel.
It should be noted that the waste liquid pumping channel provided by the invention participates in two steps of processes, specifically:
firstly, in the cell reaction process, after a reaction reagent is injected into a buffer groove from a reagent groove, the reaction reagent is pumped outwards through a waste liquid pumping channel, and the waste liquid groove, a reaction cavity and the buffer groove are communicated in sequence, so that the reaction reagent temporarily stored in the buffer groove can be pumped into the reaction cavity under the action of suction force, but the suction negative pressure needs to be specially noticed, so that the reaction reagent entering the reaction cavity is prevented from being further pumped into the waste liquid groove;
secondly, after the cell reaction is finished, the reaction waste liquid is pumped into the waste liquid groove through the waste liquid pumping channel again, and the reaction waste liquid remained in the reaction cavity is pumped into the waste liquid groove under the action of suction force.
Preferably, an electromagnetic valve is arranged at the air exhaust end of the waste liquid air exhaust channel and used for controlling the air exhaust amount.
According to the analysis, the waste liquid air exhaust channel participates in two steps of processes, the two steps of processes need to control the air exhaust amount, particularly, the air exhaust amount needs to be strictly controlled in the first step of process, and the situation that the reaction reagent in the reaction cavity is further pumped into the waste liquid tank due to overlarge negative pressure generated by air exhaust is prevented.
As a preferred technical scheme of the present invention, a product groove is further disposed on a side surface of the cell reaction plate, which is attached to the integrated gas circuit control plate, the product groove is communicated with the reaction chamber, a product pumping channel is disposed inside the integrated gas circuit control plate, the product pumping channel is communicated with the waste liquid groove, the reaction chamber, the product groove and the product pumping channel are sequentially communicated along a pumping direction, and a reaction product obtained in the reaction chamber is pumped into the product groove by pumping through the product pumping channel.
Preferably, the air exhaust end of the product air exhaust channel is provided with an electromagnetic valve, and the electromagnetic valve is used for controlling the air exhaust amount.
In the invention, the electromagnetic valve connected to the product pumping channel is mainly used for controlling the pumping amount, after the cell reaction is finished, the product tank is pumped through the product pumping channel, so that the reaction product in the reaction cavity enters the product tank, and the pumping amount needs to be strictly controlled in order to pump all the reaction product in the reaction cavity into the product tank.
As a preferred technical scheme of the present invention, a silica gel pad is sandwiched between the integrated gas circuit control board and the cell reaction plate, and the silica gel pad is provided with a through hole, and the integrated gas circuit control board and the cell reaction plate are communicated through the through hole.
As a preferred technical solution of the present invention, the electromagnetic valves are collectively disposed on a surface of the integrated gas circuit control board, which is far away from the cell reaction plate.
The invention realizes the control of the whole flow path by arranging different types of electromagnetic valves, different electromagnetic valves realize different control functions, and the traditional fluid electromagnetic valve has the problems of large occupied space, complex installation and the like when being installed and used.
Preferably, the integrated gas circuit control board is also provided with an observation window, and the reaction condition in the reaction cavity is observed through the observation window.
In a second aspect, the present invention provides a method for performing a cell reaction using the cell reaction module of the first aspect, the method comprising:
the reagent groove is filled with reaction reagent in advance, the driving gas is filled into the reagent groove through the driving gas path, and the reaction reagent in the reagent groove flows into the reaction cavity under the driving of the pressure of the driving gas to carry out cell reaction.
As a preferred technical solution of the present invention, the method specifically includes:
the method comprises the following steps that (I) driving gas is introduced into a driving gas path channel through an electromagnetic valve, the driving gas is injected into a corresponding reagent groove along an independent driving gas path channel, and a reaction reagent stored in the reagent groove is pressed into a buffer groove under the driving of the pressure of the driving gas;
(II) exhausting the waste liquid tank through a waste liquid exhaust channel, so that the reaction reagent in the buffer tank is pumped into the reaction chamber to complete reagent injection;
and (III) repeating the step (I) and the step (II) so that the reaction reagent in each reagent groove is injected into the reaction cavity to perform cell reaction.
As a preferred technical solution of the present invention, the method further comprises: after the cell reaction is finished, the waste liquid tank is pumped by the waste liquid pumping channel again, so that the waste liquid in the reaction cavity enters the waste liquid tank.
As a preferred technical solution of the present invention, the method further comprises: after the cell reaction is finished, the product tank is pumped through the product pumping channel, so that the reaction product in the reaction cavity enters the product tank.
The system refers to an equipment system, or a production equipment.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, all the driving gas channels are integrated into a whole in a manner of slotting in the integrated gas circuit control board, and part of the driving gas channels are communicated through the electromagnetic valve, so that the communication or independence among different driving gas channels is realized according to requirements through controlling a switch by an upper computer, and the driving gas is used as the driving force for injecting the reaction reagent to carry out flow path conveying of the reaction reagent. In the use, inject into drive gas in the drive gas way passageway of difference, can inject the reaction reagent of different reagent inslots into the reaction chamber one by one, the injection process of different reaction reagent is independent each other and does not influence for the flow path control process of reaction reagent is reliable and stable, whole cell reaction module simple to operate and occupation space are little.
Drawings
FIG. 1 is a schematic view of a cell reaction plate according to an embodiment of the present invention;
FIG. 2 is a plan view of a cell reaction plate according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of an integrated gas circuit control board according to an embodiment of the present invention;
FIG. 4 is a top view of an integrated air circuit control board according to an embodiment of the present invention;
wherein, 1-cell reaction plate; 2-reagent tank; 3-a buffer tank; 4-a waste liquid tank; 5-a product tank; 6-a reaction chamber; 7-integrating a gas circuit control board; 8-an electromagnetic valve; 9-observation window; 10-driving the gas circuit channel.
Detailed Description
It is to be understood that in the description of the present invention, the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be taken as limiting the present invention. Furthermore, the terms "first", "second", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first," "second," etc. may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.
It should be noted that, in the description of the present invention, unless otherwise explicitly specified or limited, the terms "disposed," "connected" and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art through specific situations.
It should be understood by those skilled in the art that the present invention necessarily includes necessary piping, conventional valves and general pump equipment for achieving the complete process, but the above contents do not belong to the main inventive points of the present invention, and those skilled in the art can select the layout of the additional equipment based on the process flow and the equipment structure, and the present invention is not particularly limited to this.
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
In a specific embodiment, the invention provides an integrated gas circuit controlled cell reaction module, the cell reaction module comprises an integrated gas circuit control board 7 (as shown in fig. 1) and a cell reaction board 1 (as shown in fig. 3) which are mutually attached, at least two mutually independent driving gas circuit channels 10 are arranged in the integrated gas circuit control board 7, reagent grooves 2 with the same number as the driving gas circuit channels 10 are arranged on the surface of one side of the cell reaction board 1 which is attached to the integrated gas circuit control board 7, and each driving gas circuit channel 10 is independently communicated with one reagent groove 2; the cell reaction plate 1 is far away from the side surface of the integrated gas circuit control plate 7 and is provided with a reaction cavity 6, the reaction cavity 6 is communicated with a reagent groove 2, a reaction reagent is injected into the reagent groove 2 in advance, a driving gas is injected into the reagent groove 2 through a driving gas circuit channel 10, and the reaction reagent in the reagent groove 2 flows into the reaction cavity 6 under the driving of the pressure of the driving gas.
All the driving gas channels 10 are integrated into a whole in a mode of slotting in the integrated gas circuit control plate 7, all the driving gas channels 10 are independent from each other and are not communicated with each other, and the driving gas is used as the driving force for injecting the reaction reagent to carry out flow path conveying of the reaction reagent. In the use, inject into drive gas in different drive gas way passageways 10, can inject the reaction reagent of different reagent grooves 2 into reaction chamber 6 one by one, the injection process of different reaction reagent is independent each other and does not influence for the flow path control process of reaction reagent is reliable and stable, whole cell reaction module simple to operate and occupation space are little.
In the present embodiment, the opening manner of the driving gas passage 10 adopts the following two schemes:
the first scheme is as follows: the integrated gas circuit control plate 7 is of an integrated structure, and the driving gas circuit channel 10 is directly arranged in the integrated gas circuit control plate 7 in a pouring, drilling or additive manufacturing mode and the like;
scheme II: the integrated gas circuit control board 7 is of a split structure and is formed by laminating and attaching a gas circuit upper board and a gas circuit lower board, a gas circuit groove is formed in an attaching surface between the gas circuit upper board and/or the gas circuit lower board, and after the gas circuit upper board and the gas circuit lower board are attached, the gas circuit groove is sealed to form a driving gas circuit channel 10; it can be understood that, in the present embodiment, the air channel groove may be opened on the lower surface of the air channel upper plate or the upper surface of the air channel lower plate, or may be opened on both the lower surface of the air channel upper plate and the upper surface of the air channel lower plate, as shown in fig. 4, the air channel groove is opened on the lower surface of the air channel upper plate, and the air channel lower plate is not shown in the figure, but it can also be understood that, after the air channel lower plate and the air channel upper plate are relatively attached, the air channel groove shown in fig. 4 is closed by the air channel lower plate to form the driving air channel 10.
It should be noted that, the cell reaction plate 1 is also provided with a micro flow channel for flowing the reaction reagent, the manner of providing the micro flow channel can be referred to the above description about the driving gas channel 10, that is, the cell reaction plate 1 can be an integrated structure or a split structure, when the cell reaction plate 1 is an integrated structure, the micro flow channel is directly provided inside the cell reaction plate 1 by casting, drilling or additive manufacturing; certainly, the cell reaction plate 1 may be a split structure formed by laminating and attaching a reaction upper plate and a reaction lower plate, a joint surface of the reaction upper plate and/or the reaction lower plate is provided with a flow channel groove, and the flow channel groove is closed to form a microchannel after the reaction upper plate and the reaction lower plate are joined, as shown in fig. 2, the flow channel groove is provided on a lower surface of the reaction upper plate, the reaction lower plate is not shown in the figure, but it can also be understood that, after the reaction lower plate and the reaction upper plate are joined relatively, the flow channel groove shown in fig. 2 is closed by the reaction lower plate to form the microchannel.
Cell reaction plate 1 still is provided with dashpot 3 with one side surface of the laminating of integrated gas circuit control panel 7, and reagent groove 2 and reaction chamber 6 communicate dashpot 3 independently respectively, along the flow direction of drive gas, and drive gas way passageway 10, reagent groove 2 and dashpot 3 communicate in proper order, and drive gas lets in reagent groove 2 through drive gas way passageway 10, and the reaction agent who stores in each reagent groove 2 gets into dashpot 3 one by one to pour into reaction chamber 6 through dashpot 3. An electromagnetic valve 8 is arranged at the air inlet end of the driving air channel 10, and the electromagnetic valve 8 is used for controlling the introduction amount of the driving air. In the cell reaction process, the reaction reagent stored in the reagent tank 2 is pressed into the buffer tank 3 under the pressure drive of the drive gas, and the flow of the drive gas is adjusted by the electromagnetic valve 8, so that the injection amount of the reaction reagent into the buffer tank 3 is changed.
Cell reaction plate 1 still is provided with waste liquid groove 4 with one side surface of the laminating of integrated gas circuit control panel 7, waste liquid groove 4 and reaction chamber 6 intercommunication, integrated gas circuit control panel 7 is inside to have seted up waste liquid pumping channel, waste liquid pumping channel and waste liquid groove 4 intercommunication, along the direction of bleeding, reaction chamber 6, waste liquid groove 4 and waste liquid pumping channel communicate in proper order, bleed through waste liquid pumping channel, waste liquid suction waste liquid groove 4 after finishing reacting in the reaction chamber 6. The exhaust end of the waste liquid exhaust channel is provided with an electromagnetic valve 8, and the electromagnetic valve 8 is used for controlling the exhaust amount.
It should be noted that the waste liquid pumping channel participates in two steps of processes, specifically:
firstly, in the cell reaction process, after a reaction reagent is injected into the buffer groove 3 from the reagent groove 2, the reaction reagent is pumped outwards through the waste liquid pumping channel, and as the waste liquid groove 4, the reaction cavity 6 and the buffer groove 3 are sequentially communicated, the reaction reagent temporarily stored in the buffer groove 3 can be pumped into the reaction cavity 6 under the action of suction force, but the negative pressure of pumping needs to be specially noticed at the moment, so that the reaction reagent entering the reaction cavity 6 is prevented from being further pumped into the waste liquid groove 4;
secondly, after the cell reaction is finished, the reaction waste liquid is pumped into the waste liquid tank 4 by the waste liquid pumping channel again, and the reaction waste liquid remained in the reaction cavity 6 is pumped into the waste liquid tank by the action of suction force.
The cell reaction plate 1 still is provided with the result groove 5 with one side surface of the laminating of integrated gas circuit control panel 7, and result groove 5 and reaction chamber 6 intercommunication have been seted up to integrated gas circuit control panel 7 inside and have been offered the result air exhaust passage, and the result air exhaust passage communicates with waste liquid groove 4, and along the direction of bleeding, reaction chamber 6, result groove 5 and result air exhaust passage communicate in proper order, bleed through the result air exhaust passage, with the reaction product suction that obtains in the reaction chamber 6 in the result groove 5. And the air exhaust end of the product air exhaust channel is provided with an electromagnetic valve 8, and the electromagnetic valve 8 is used for controlling the air exhaust amount. After the cell reaction, bleed to product groove 5 through the product bleed passage for reaction product in the reaction chamber 6 gets into product groove 5, in order to with the whole suction of reaction product in the reaction chamber 6 product groove 5, need carry out the close control to the air extraction volume, sets up solenoid valve 8 through the end of bleeding at waste liquid bleed passage, combines control module to carry out automatic control to the air extraction volume.
A silica gel pad is clamped between the integrated gas circuit control plate 7 and the cell reaction plate 1, a through hole is formed in the silica gel pad, and the integrated gas circuit control plate 7 and the cell reaction plate 1 are communicated through the through hole. The electromagnetic valves 8 are collectively arranged on the surface of one side of the integrated gas circuit control plate 7 far away from the cell reaction plate 1. The control of whole flow path has been realized through the solenoid valve 8 that sets up the different grade type, different solenoid valves 8 have realized different control functions, among the cell reaction module that this embodiment provided, installation position has been reserved for solenoid valve 8 on integrated gas circuit control panel 7 surface, the installation and the dismantlement of the solenoid valve 8 of being convenient for, install solenoid valve 8 integration in integrated gas circuit control panel 7 surface simultaneously, the inside flow path of fluid accessible integrated gas circuit control panel 7 realizes the control to whole flow path. An observation window 9 is also arranged on the integrated gas circuit control board 7, and the reaction condition in the reaction cavity 6 is observed through the observation window 9.
In another embodiment, the present invention provides a method for performing a cell reaction using the cell reaction module according to the above embodiment, the method comprising:
(1) driving gas is introduced into a driving gas path channel 10 through an electromagnetic valve 8, the driving gas is injected into a corresponding reagent groove 2 along the independent driving gas path channel 10, and a reaction reagent stored in the reagent groove 2 is pressed into a buffer groove 3 under the driving of the pressure of the driving gas;
(2) pumping the waste liquid tank 4 through a waste liquid pumping channel so that the reaction reagent in the buffer tank 3 is pumped into the reaction chamber to complete reagent injection;
(3) repeating the step (1) and the step (2) to inject the reaction reagent in each reagent groove 2 into the reaction cavity 6 for cell reaction;
(4) after the cell reaction is finished, the waste liquid tank 4 is pumped through the waste liquid pumping channel again, so that the waste liquid in the reaction cavity 6 enters the waste liquid tank 4; the product tank 5 is evacuated through the product evacuation passage, so that the reaction product in the reaction chamber 6 enters the product tank 5.
Application example
The present application example provides a specific cell reaction method, which comprises:
(1) various reaction reagents are injected into each reagent groove 2 in the cell reaction plate 1 in advance, wherein the reaction reagents comprise 9 reagents of cell buffer solution, chip surface processing solvent, cell lysate, reverse transcription reagent, reverse transcription cleaning solution and label magnetic beads;
(2) driving gas is introduced into a driving gas path channel 10 through an electromagnetic valve 8, the driving gas is injected into a corresponding reagent groove 2 along the independent driving gas path channel 10, and a reaction reagent stored in the reagent groove 2 is pressed into a buffer groove 3 under the driving of the pressure of the driving gas;
(3) pumping the waste liquid tank 4 through a waste liquid pumping channel so that the reaction reagent in the buffer tank 3 is pumped into the reaction chamber to complete reagent injection;
(4) repeating the step (2) and the step (3), so that the reaction reagent in each reagent groove 2 is injected into the reaction cavity 6, the temperature of the cell reaction is controlled at 42 ℃, and the DNA product is obtained after the reaction is carried out for 1.5 h;
(5) after the cell reaction is finished, the waste liquid tank 4 is pumped through the waste liquid pumping channel again, so that the waste liquid in the reaction cavity 6 enters the waste liquid tank 4; the product tank 5 is evacuated through the product evacuation passage, so that the reaction product in the reaction chamber 6 enters the product tank 5.
The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.
Claims (10)
1. The cell reaction module is characterized by comprising an integrated gas circuit control plate and a cell reaction plate which are mutually attached, wherein at least two mutually independent driving gas circuit channels are formed in the integrated gas circuit control plate, the surface of one side of the cell reaction plate, which is attached to the integrated gas circuit control plate, is provided with reagent grooves with the same number as the driving gas circuit channels, and each driving gas circuit channel is independently communicated with one reagent groove; the cell reaction plate is characterized in that a reaction cavity is formed in the surface of one side, away from the integrated gas circuit control plate, of the cell reaction plate, the reaction cavity is communicated with a reagent groove, a reaction reagent is injected into the reagent groove in advance, a driving gas is injected into the reagent groove through a driving gas circuit channel, and the reaction reagent in the reagent groove flows into the reaction cavity under the driving of the pressure of the driving gas.
2. The cell reaction module according to claim 1, wherein a buffer groove is further provided on a side surface of the cell reaction plate to which the integrated gas circuit control plate is attached, the reagent groove and the reaction chamber are respectively and independently communicated with the buffer groove, the driving gas circuit channel, the reagent groove and the buffer groove are sequentially communicated along a flow direction of driving gas, the driving gas is introduced into the reagent groove through the driving gas circuit channel, and the reaction reagents stored in the reagent grooves enter the buffer groove one by one and are injected into the reaction chamber through the buffer groove;
preferably, an electromagnetic valve is arranged at the air inlet end of the driving air channel and used for controlling the introduction amount of the driving air.
3. The cell reaction module according to claim 1 or 2, wherein a waste liquid tank is further disposed on a side surface of the cell reaction plate, which is attached to the integrated gas circuit control plate, the waste liquid tank is communicated with the reaction chamber, a waste liquid pumping channel is disposed inside the integrated gas circuit control plate, the waste liquid pumping channel is communicated with the waste liquid tank, the reaction chamber, the waste liquid tank and the waste liquid pumping channel are sequentially communicated along a pumping direction, and the waste liquid after the reaction in the reaction chamber is pumped into the waste liquid tank by pumping air through the waste liquid pumping channel;
preferably, an electromagnetic valve is arranged at the air exhaust end of the waste liquid air exhaust channel and used for controlling the air exhaust amount.
4. The cell reaction module according to any one of claims 1 to 3, wherein a product groove is further disposed on a side surface of the cell reaction plate, which is attached to the integrated gas circuit control plate, the product groove is communicated with the reaction chamber, a product pumping channel is disposed inside the integrated gas circuit control plate, the product pumping channel is communicated with the waste liquid groove, the reaction chamber, the product groove and the product pumping channel are sequentially communicated along a pumping direction, and a reaction product obtained in the reaction chamber is pumped into the product groove by pumping through the product pumping channel;
preferably, the air exhaust end of the product air exhaust channel is provided with an electromagnetic valve, and the electromagnetic valve is used for controlling the air exhaust amount.
5. The cell reaction module according to any one of claims 1 to 4, wherein a silica gel pad is interposed between the integrated gas circuit control plate and the cell reaction plate, and the silica gel pad is provided with a through hole for communicating the integrated gas circuit control plate and the cell reaction plate through a passage through hole.
6. A cell reaction module according to any one of claims 2-5, wherein the electromagnetic valves are collectively disposed on a surface of the integrated gas circuit control board away from the cell reaction plate;
preferably, the integrated gas circuit control board is also provided with an observation window, and the reaction condition in the reaction cavity is observed through the observation window.
7. A method for performing a cell reaction using the cell reaction module according to any one of claims 1 to 6, the method comprising:
the reagent groove is filled with reaction reagent in advance, the driving gas is filled into the reagent groove through the driving gas path, and the reaction reagent in the reagent groove flows into the reaction cavity under the driving of the pressure of the driving gas to carry out cell reaction.
8. The method according to claim 7, characterized in that the method comprises in particular:
the method comprises the following steps that (I) driving gas is introduced into a driving gas path channel through an electromagnetic valve, the driving gas is injected into a corresponding reagent groove along an independent driving gas path channel, and a reaction reagent stored in the reagent groove is pressed into a buffer groove under the driving of the pressure of the driving gas;
(II) exhausting the waste liquid tank through a waste liquid exhaust channel, so that the reaction reagent in the buffer tank is pumped into the reaction chamber to complete reagent injection;
and (III) repeating the step (I) and the step (II) so that the reaction reagent in each reagent groove is injected into the reaction cavity to perform cell reaction.
9. The method of claim 7 or 8, further comprising: after the cell reaction is finished, the waste liquid tank is pumped by the waste liquid pumping channel again, so that the waste liquid in the reaction cavity enters the waste liquid tank.
10. The method according to any one of claims 7-9, further comprising: after the cell reaction is finished, the product tank is pumped through the product pumping channel, so that the reaction product in the reaction cavity enters the product tank.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110534947.0A CN113234583A (en) | 2021-05-17 | 2021-05-17 | Cell reaction module integrated with gas circuit control and method for carrying out cell reaction by adopting cell reaction module |
| CN202180092485.XA CN116867889A (en) | 2020-12-02 | 2021-12-02 | Reagent exchange method, device and system |
| PCT/CN2021/135159 WO2022117053A1 (en) | 2020-12-02 | 2021-12-02 | Reagent exchange methods, devices, and systems |
| US18/255,569 US20240033727A1 (en) | 2020-12-02 | 2021-12-02 | Reagent exchange methods, devices, and systems |
| EP21900079.1A EP4256030A1 (en) | 2020-12-02 | 2021-12-02 | Reagent exchange methods, devices, and systems |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110534947.0A CN113234583A (en) | 2021-05-17 | 2021-05-17 | Cell reaction module integrated with gas circuit control and method for carrying out cell reaction by adopting cell reaction module |
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| CN113234583A true CN113234583A (en) | 2021-08-10 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110534947.0A Pending CN113234583A (en) | 2020-12-02 | 2021-05-17 | Cell reaction module integrated with gas circuit control and method for carrying out cell reaction by adopting cell reaction module |
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| WO2022117053A1 (en) * | 2020-12-02 | 2022-06-09 | Suzhou Singleron Biotechnologies Co., Ltd. | Reagent exchange methods, devices, and systems |
| CN114887674A (en) * | 2022-04-26 | 2022-08-12 | 北京百迈客生物科技有限公司 | Micro-droplet generating device |
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| CN108117969A (en) * | 2016-11-28 | 2018-06-05 | 中国科学院大连化学物理研究所 | Single celled micro-fluidic chip of a kind of high throughput automatic capture and preparation method thereof |
| CN109136352A (en) * | 2018-08-10 | 2019-01-04 | 深圳先进技术研究院 | Sample processing device, micro-fluidic chip and application before a kind of unicellular sequencing |
| CN112322486A (en) * | 2020-10-23 | 2021-02-05 | 苏州新格元生物科技有限公司 | Single cell nucleic acid processing instrument |
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| CN108117969A (en) * | 2016-11-28 | 2018-06-05 | 中国科学院大连化学物理研究所 | Single celled micro-fluidic chip of a kind of high throughput automatic capture and preparation method thereof |
| CN109136352A (en) * | 2018-08-10 | 2019-01-04 | 深圳先进技术研究院 | Sample processing device, micro-fluidic chip and application before a kind of unicellular sequencing |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2022117053A1 (en) * | 2020-12-02 | 2022-06-09 | Suzhou Singleron Biotechnologies Co., Ltd. | Reagent exchange methods, devices, and systems |
| CN114887674A (en) * | 2022-04-26 | 2022-08-12 | 北京百迈客生物科技有限公司 | Micro-droplet generating device |
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