CN115353972A - Cell sample preparation device and preparation method thereof - Google Patents
Cell sample preparation device and preparation method thereof Download PDFInfo
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- CN115353972A CN115353972A CN202110535870.9A CN202110535870A CN115353972A CN 115353972 A CN115353972 A CN 115353972A CN 202110535870 A CN202110535870 A CN 202110535870A CN 115353972 A CN115353972 A CN 115353972A
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Abstract
The invention provides a cell sample preparation device and a preparation method thereof, wherein the preparation device comprises a rack, a reaction unit is arranged on the rack, the reaction unit comprises an air path control plate and a heating plate, and a reaction chip is clamped between the air path control plate and the heating plate; at least two mutually independent driving air channel channels are arranged in the air channel control plate; the reaction chip comprises a carrying platform and a cell reaction plate which are arranged in an attaching mode, wherein reagent grooves with the same number as that of the driving air channel are formed in the surface of one side, attached to the air channel control plate, of the cell reaction plate, and each driving air channel is independently communicated with one reagent groove; a reaction cavity is formed in the surface of one side, attached to the carrier, of the cell reaction plate, and the reagent grooves are independently connected with the reaction cavity; the reagent groove is filled with reaction reagent in advance, and driving gas is injected into the reagent groove through the driving gas path, so that the reaction reagent in the reagent groove is pressed into the reaction cavity for reaction. The invention has the characteristics of simple structure, simple and easy operation, small occupied area, strong adaptability and the like.
Description
Technical Field
The invention belongs to the technical field of cell sample preparation, relates to a cell sample preparation device, and particularly relates to a cell sample preparation device and a preparation method thereof.
Background
With the maturation of sequencing technology, high-throughput single-cell sequencing is gradually emphasized, the single-cell sequencing technology mainly comprises the steps of marking a sample target cell population, marking molecular labels with different sequences on one cell, and then performing sequencing analysis on the whole sample to obtain the difference of the cell heterogeneity of the sample, and the method has wide application in clinic and treatment.
However, in a series of processes from processing a cell sample, labeling the cell sample with a molecular tag, and then performing reverse transcription, a series of instruments and equipment and skilled operators are required for a conventional laboratory to perform the operation, and the whole operation flow is usually completed in a period of half a day to a day.
At present, automatic instruments for high-throughput single-cell preparation in the market only carry out molecular label labeling, but have the problems of complex structure, large occupied area, complex operation and the like.
CN107354093A discloses a cell preparation apparatus comprising: the device comprises a frame, a positioning device and a control device, wherein the frame is provided with at least two stations; the cell separation and culture device is used for separating and culturing cells; wherein, the cell separation culture device is arranged on the frame. According to the cell preparation equipment, at least two stations are arranged, and the cell separation culture devices correspond to the stations one by one, so that at least two cells can be prepared simultaneously, and the cell preparation efficiency is effectively improved; meanwhile, at least two identical cells are prepared, so that the deviation between the two cells caused by different equipment is effectively avoided; at least two different cells are prepared without purchasing additional equipment, thereby reducing the number of required equipment. But its structure is complicated and the occupied area is large.
CN107636142A discloses an automatic cell culture device and its operation method, comprising: an incubator which accommodates at least one container for culturing cells; a microscope for observing the state of the cells in the container; a robot for moving the position of the container; a liquid handler for flowing liquid into or out of the container; and a control device for controlling operation of at least one of the incubator, the microscope, the robot and the liquid handler. The position of the container is moved by the manipulator, so that the problems of complex structure, inconvenient operation and the like exist.
CN106367343A discloses a full-automatic intelligent cell culture device and a control method thereof, which comprises a controller, a first adjusting mechanism, a second adjusting mechanism, a third adjusting mechanism, a culture frame, a first control door, a second control door, a first liquid transferring mechanism, a second liquid transferring mechanism, a waste liquid collecting mechanism, a bottle opening and closing mechanism and a liquid adding mechanism, wherein at least three cavities are arranged in a storage cavity, and the storage cavity comprises a storage cavity, a transition cavity and a preheating cavity. The device is convenient, rapid and accurate in liquid changing, has no influence on cell growth, and is convenient to preheat and high in preheating speed. But still has the problems of complex structure, inconvenient operation and floor space reaching.
The existing cell sample preparation devices all have the problems of complex structure, large occupied area, inconvenient operation and the like, so that the cell sample preparation devices can be ensured to be operated simply and conveniently and meet the requirement of reverse transcription operation of cell samples under the condition of simple structure and small occupied area, the operation threshold of operators is reduced, and the cell sample preparation devices become the problems which need to be solved urgently at present.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a cell sample preparation device and a preparation method thereof, wherein a gas circuit control plate is combined with a reaction chip for use, a driving gas is used for controlling the addition of a reaction reagent, so that the addition amount of the reaction reagent is effectively controlled, the reaction reagent is not polluted, and the reverse transcription of a cell sample is further controlled by a heating plate, so that the cell sample preparation device has the characteristics of simple structure, small occupied area, convenience in operation, strong adaptability and the like.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a cell sample preparation device, which comprises a rack, wherein a reaction unit is arranged on the rack, the reaction unit comprises an air path control plate and a heating plate, and a reaction chip is clamped between the air path control plate and the heating plate; at least two mutually independent driving air channel channels are arranged in the air channel control plate; the reaction chip comprises a carrying platform and a cell reaction plate which are arranged in a bonding mode, wherein reagent grooves with the same number as that of the driving gas path channels are formed in the surface of one side, which is bonded with the gas path control plate, of the cell reaction plate, and each driving gas path channel is independently communicated with one reagent groove; a reaction cavity is formed in the surface of one side, attached to the carrier, of the cell reaction plate, and the reagent grooves are independently connected with the reaction cavity; the reagent groove is filled with a reaction reagent in advance, and driving gas is injected into the reagent groove through the driving gas path, so that the reaction reagent in the reagent groove is pressed into the reaction cavity.
According to the invention, the reagent groove is arranged on the reaction chip, and the driving gas path channel on the gas path control plate is combined to inject gas into the reagent groove, so that the reaction reagent is injected into the reaction cavity to perform cell reaction, the injection of the reaction reagent is controlled by using the driving gas, and the reaction is actually injected into the reagent groove, so that different actual batch injections or common injections are realized, so that the quantitative injection of different reaction reagents is realized, the operation difficulty of operators is effectively simplified, and the structure of the reaction unit is simplified due to the matching of the gas path control plate and the reaction chip; furthermore, the reflecting chip is heated by arranging the heating plate, so that the invention has the function of reverse transcription, and has the characteristics of simple structure, simple and easy operation, small occupied area, strong adaptability and the like.
As a preferred technical scheme of the invention, the cell reaction plate is also provided with a product groove and a waste liquid groove on the surface of one side of the reagent groove, the gas circuit control plate is also internally provided with at least two gas pumping channels, and the product groove and the waste liquid groove are respectively and independently connected with the gas pumping channels.
Preferably, the product tank and the waste liquid tank are respectively and independently connected with the reaction cavity.
Preferably, the product tank and the waste liquid tank are both connected with an air suction electromagnetic valve through the air suction channel.
The reaction chamber is provided with a product tank and a waste liquid tank which are respectively communicated with the air exhaust electromagnetic valve, so that a cell sample in the reaction chamber can be sucked, and in addition, the reaction reagent in the buffer tank is further helped to enter the reaction chamber by exhausting the waste liquid tank; after the reaction is completed, the cell sample after the reaction is collected to the product tank by pumping air to the product tank.
As a preferable technical scheme of the invention, the reagent tanks are connected with a gas injection electromagnetic valve through the driving gas channel.
Preferably, the air exhaust solenoid valve and the air injection solenoid valve are both arranged on the same side surface of the air path control plate in a centralized manner.
According to the invention, the air exhaust electromagnetic valve and the air injection electromagnetic valve are arranged on the same side surface of the air circuit control board in a centralized manner, so that the integration level of the device is improved, the problem of messy pipelines is avoided, and the floor area of the device is reduced.
In a preferred embodiment of the present invention, the cell reaction plate has a buffer groove on a surface of one side of the reagent groove, the reagent groove and the reaction chamber are independently communicated with the buffer groove, and the reagent groove, the buffer groove and the reaction chamber are sequentially connected in a flow direction of the reaction reagent.
Preferably, a silica gel pad is arranged between the cell reaction plate and the air channel control plate, and a hole corresponding to the outlet of the driving air channel on the air channel control plate is formed in the silica gel pad.
As a preferred technical solution of the present invention, the air path control plate includes an upper air path plate and a lower air path plate stacked together.
Preferably, the joint surface of the air path lower plate and the air path upper plate is provided with at least one air path groove, the air path upper plate is jointed with the air path lower plate, and the air path groove is closed and sealed to form the driving air path channel and the air exhaust channel.
In a preferred embodiment of the present invention, at least one independent reagent flow channel is further formed on a surface of the cell reaction plate on the side where the stage is attached, and the reagent flow channel forms a reagent flow channel after the stage is attached and sealed to the cell reaction plate.
Preferably, the reagent reservoirs are independently connected to the buffer reservoirs through the reagent flow channels, respectively.
Preferably, the buffer reservoirs are independently connected to the reaction chamber through the reagent flow channel.
Preferably, the product tank and the waste tank are both independently connected to the reaction chamber through the reagent flow channel.
As a preferred technical solution of the present invention, a control unit is disposed at the bottom of the reaction unit, the control unit is separately and electrically connected to the heating plate, the gas injection solenoid valve and the gas extraction solenoid valve, and the control unit separately and independently controls the start of the heating plate, the start of the gas injection solenoid valve and the start of the gas extraction solenoid valve.
As a preferred technical scheme of the present invention, the gas injection solenoid valve and the gas extraction solenoid valve are both connected to a gas pump assembly, and the gas pump assembly is used for providing the pressure of the gas in the gas injection solenoid valve and the gas extraction solenoid valve.
Preferably, the air pump assembly is located below the reaction unit.
Preferably, the air pump assembly is arranged side by side with the control unit.
According to the invention, the control unit and the air pump assembly are integrally arranged at the bottom of the reaction unit, so that the floor area of the device is further reduced, and the integration level of the device is improved.
In a second aspect, the present invention also provides a method for preparing a cell sample using the cell sample preparation device according to the first aspect, the method comprising:
injecting a reaction reagent into the reagent cavity, placing the reaction chip between the air circuit control plate and the heating plate, injecting a driving gas into the reagent groove through the driving air circuit channel, pressing the reaction reagent in the reagent groove into the reaction cavity for cell reaction, and starting the heating plate to heat the reaction cavity for reverse transcription to prepare the cell sample.
It should be noted that the present invention does not specifically require or limit the kind of the reaction reagent, and those skilled in the art can appropriately select the kind and the amount of the reaction reagent according to the kind of the cell sample to be prepared.
As a preferred technical scheme of the invention, the preparation method specifically comprises the following steps:
after injecting a reaction reagent into the reagent cavity, placing the reaction chip between the gas path control plate and the heating plate, controlling the gas injection electromagnetic valve to be started by the control unit, injecting gas into the reagent cavity to press the reaction reagent into the buffer cavity, controlling the gas extraction electromagnetic valve of the waste liquid tank to be started by the control unit, and sucking the reaction reagent in the buffer cavity into the reaction cavity by the gas extraction of the waste liquid tank to perform cell reaction;
(II) repeating the operation of the step (I) at least once, wherein the control unit controls the heating plate to heat and reversely transcribe the reaction cavity to prepare the cell sample;
and (III) the control unit controls the starting of an air exhaust electromagnetic valve of the product tank to suck the cell sample in the reaction cavity into the product tank.
The numerical ranges set forth herein include not only the recited values but also any values between the recited numerical ranges not enumerated herein, and are not intended to be exhaustive or otherwise clear from the intended disclosure of the invention in view of brevity and clarity.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the reagent groove is arranged on the reaction chip, and the driving gas path channel on the gas path control plate is combined to inject gas into the reagent groove, so that the reaction reagent is injected into the reaction cavity to perform cell reaction, the injection of the reaction reagent is controlled by using the driving gas, and the reaction is actually injected into the reagent groove, so that different actual batch injections or common injections are realized, so that the quantitative injection of different reaction reagents is realized, the operation difficulty of operators is effectively simplified, and the structure of the reaction unit is simplified due to the matching of the gas path control plate and the reaction chip; furthermore, the reflecting chip is heated by arranging the heating plate, so that the invention has the function of reverse transcription, and has the characteristics of simple structure, simple and easy operation, small occupied area, strong adaptability and the like.
Drawings
FIG. 1 is a schematic structural view of a cell sample preparation device according to an embodiment of the present invention;
FIG. 2 is a schematic top view of a cell reaction plate according to an embodiment of the present invention;
FIG. 3 is a schematic structural diagram of a bonding surface of a cell reaction plate and a carrier according to an embodiment of the present invention;
fig. 4 is a schematic diagram of an internal structure of an air path control plate according to an embodiment of the present invention.
Wherein, 1-heating plate; 2-a gas circuit control panel; 3-an air pump assembly; 4-a control unit; 5-reagent tank; 6-a product tank; 7-a waste liquid tank; 8-a reaction chamber; 9-reagent flow cell; 10-an air exhaust channel; 11-air exhaust electromagnetic valve; 12-a drive gas path channel; 13-gas injection solenoid valve.
Detailed Description
It is to be understood that in the description of the present invention, the terms "central," "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in an orientation or positional relationship indicated in the drawings for convenience in describing the present invention and to simplify the description, but are not intended to indicate or imply that the device or element so referred to must have a particular orientation, be constructed in a particular orientation, and be operated in a particular manner and therefore are not to be construed as limiting the invention.
It should be noted that, unless explicitly stated or limited otherwise, the terms "disposed," "connected" and "connected" in the description of the present invention are to be construed broadly and may include, for example, a fixed connection, a detachable connection, or an integral connection; 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.
The technical solution of the present invention is further explained by the following embodiments.
In one embodiment, the present invention provides a cell sample preparation apparatus, as shown in fig. 1, fig. 2, fig. 3 and fig. 4, the preparation apparatus includes a frame, a reaction unit is disposed on the frame, the reaction unit includes a gas circuit control board 2 and a heating plate 1, and a reaction chip is sandwiched between the gas circuit control board 2 and the heating plate 1; at least two mutually independent driving air channel channels 12 are arranged in the air channel control plate 2; the reaction chip comprises a carrying platform and a cell reaction plate which are arranged in an attaching mode, the surface of one side, attached to the air channel control plate 2, of the cell reaction plate is provided with reagent grooves 5 the number of which is the same as that of the driving air channel 12, and each driving air channel 12 is independently communicated with one reagent groove 5; a reaction cavity 8 is formed in the surface of one side, which is attached to the carrier, of the cell reaction plate, and the reagent tanks 5 are independently connected with the reaction cavity 8; the reagent tank 5 is filled with a reaction reagent in advance, and the driving gas is injected into the reagent tank 5 through the driving gas path 12 to press the reaction reagent in the reagent tank 5 into the reaction chamber 8.
According to the invention, the reagent tank 5 is arranged on the reaction chip, and the driving gas path channel 12 on the gas path control plate 2 is combined to inject gas into the reagent tank 5, so that the reaction reagent is injected into the reaction cavity 8 for cell reaction, the injection of the reaction reagent is controlled by using the driving gas, and the reaction is actually injected into the reagent tank 5, so that different actual batch injections or common injections are realized, so that the quantitative injection of different reaction reagents is realized, the operation difficulty of operators is effectively simplified, and the structure of the reaction unit is simplified due to the matching of the gas path control plate 2 and the reaction chip; furthermore, the heating plate 1 is arranged to heat the reflecting chip, so that the reverse transcription reaction device has the reverse transcription function, and has the characteristics of simple structure, simplicity and easiness in operation, small occupied area, strong adaptability and the like.
Furthermore, the cell reaction plate is also provided with a product groove 6 and a waste liquid groove 7 on the surface of one side of the reagent groove 5, at least two air pumping channels 10 are also arranged in the air path control plate 2, and the product groove 6 and the waste liquid groove 7 are respectively and independently connected with the air pumping channels 10; the product tank 6 and the waste liquid tank 7 are respectively and independently connected with the reaction cavity 8; the product tank 6 and the waste liquid tank 7 are both connected with an air exhaust electromagnetic valve 11 through the air exhaust channel 10.
The invention can absorb the cell sample in the reaction cavity 8 by arranging the product groove 6 and the waste liquid groove 7 and respectively communicating the air exhaust electromagnetic valve 11, and in addition, the reaction reagent in the buffer groove is further helped to enter the reaction cavity 8 by exhausting the waste liquid groove 7; after the reaction is completed, the cell sample after the reaction is collected in the product tank 6 by evacuating the product tank 6.
Further, the reagent tanks 5 are connected with a gas injection electromagnetic valve 13 through the driving gas channel 12; the air exhaust electromagnetic valve 11 and the air injection electromagnetic valve 13 are both arranged on the same side surface of the air path control plate 2 in a centralized manner. According to the invention, the air exhaust electromagnetic valve 11 and the air injection electromagnetic valve 13 are intensively arranged on the same side surface of the air path control plate 2, so that the integration level of the device is improved, the problem of disordered pipelines is avoided, and the floor area of the device is reduced.
Further, the cell reaction plate is provided with a buffer groove on the surface of one side of the reagent groove 5, the reagent groove 5 and the reaction chamber 8 are respectively and independently communicated with the buffer groove, and the reagent groove 5, the buffer groove and the reaction chamber 8 are sequentially connected along the flowing direction of the reaction reagent.
Furthermore, a silica gel pad is arranged between the cell reaction plate and the air channel control plate 2, and a hole corresponding to the outlet position of the driving air channel 12 on the air channel control plate 2 is arranged on the silica gel pad.
Further, the air path control plate 2 includes an upper air path plate and a lower air path plate which are stacked. At least one air channel groove is formed in the joint surface of the air channel lower plate and the air channel upper plate, the air channel upper plate is jointed with the air channel lower plate, and the air channel groove is closed and sealed to form the driving air channel 12 and the air exhaust channel 10.
Furthermore, at least one independent reagent flow channel 9 is formed on the side surface of the cell reaction plate, which is attached to the carrier, and after the carrier and the cell reaction plate are attached and sealed, the flow channel forms a reagent flow channel. The reagent reservoirs 5 are independently connected to the buffer reservoirs through reagent flow channels, respectively. The buffer reservoirs are independently connected to the reaction chambers 8 through the reagent flow channels. The product tank 6 and the waste liquid tank 7 are independently connected with the reaction chamber 8 through a reagent flow channel.
Further, the bottom of the reaction unit is provided with a control unit 4, the control unit 4 is respectively and independently electrically connected with the heating plate 1, the gas injection electromagnetic valve 13 and the gas exhaust electromagnetic valve 11, and the control unit 4 respectively and independently controls the starting of the heating plate 1, the starting of the gas injection electromagnetic valve 13 and the starting of the gas exhaust electromagnetic valve 11.
Further, the gas injection solenoid valve 13 and the gas exhaust solenoid valve 11 are both connected to an air pump assembly 3, and the air pump assembly 3 is used for providing the pressure of the gas in the gas injection solenoid valve 13 and the gas exhaust solenoid valve 11. The air pump assembly 3 is located below the reaction unit and is arranged alongside the control unit 4. According to the invention, the control unit 4 and the air pump assembly 3 are integrally arranged at the bottom of the reaction unit, so that the floor area of the device is further reduced, and the integration level of the device is improved.
In another embodiment, the present invention further provides a preparation method for preparing a cell sample by using the above cell sample preparation device, wherein the preparation method specifically comprises the following steps:
after injecting a reaction reagent into a reagent cavity, placing a reaction chip between a gas circuit control plate 2 and a heating plate 1, controlling a gas injection electromagnetic valve 13 to start by a control unit 4, injecting gas into the reagent cavity to press the reaction reagent into a buffer cavity, controlling a gas extraction electromagnetic valve 11 of a waste liquid tank 7 to start by the control unit 4, and sucking the reaction reagent in the buffer cavity into a reaction cavity 8 by the waste liquid tank 7 through gas extraction to carry out cell reaction;
(II) repeating the operation of the step (I) at least once, wherein the control unit 4 controls the heating plate 1 to heat and reversely transcribe the reaction cavity 8 to prepare the cell sample;
(III) the control unit 4 controls the suction solenoid valve 11 of the product tank 6 to start, and the cell sample in the reaction chamber 8 is sucked into the product tank 6.
Application example
The present application example provides a method for preparing a cell sample by using the cell sample apparatus provided in one embodiment, the method specifically includes the following steps:
reagents including cell buffer solution, chip surface treatment solvent, cell lysate, reverse transcription reagent, reverse transcription cleaning solution, label magnetic beads and the like are respectively injected into a reagent cavity, a reaction chip is placed between a gas circuit control plate 2 and a heating plate 1, a control unit 4 controls an air injection electromagnetic valve 13 to be started, air is injected into the reagent cavity to respectively press the reagents into the buffer cavity, the control unit 4 controls an air extraction electromagnetic valve 11 of a waste liquid groove 7 to be started, and the waste liquid groove 7 extracts air to suck reaction reagents in the buffer groove into a reaction cavity 8 for cell reaction;
(II) the control unit 4 controls the heating plate 1 to heat and reversely transcribe the reaction cavity 8, wherein the heating temperature is 42 ℃, and the cell sample is prepared; the control unit 4 controls the air-bleed solenoid valve 11 of the product tank 6 to be activated, and sucks the cell sample in the reaction chamber 8 into the product tank 6.
The time for the whole sample preparation process was 1.5h.
Through the application example, the reagent groove 5 is arranged on the reaction chip, and the driving gas path channel 12 on the gas path control plate 2 is combined to inject gas into the reagent groove 5, so that the reaction reagent is injected into the reaction cavity 8 for cell reaction, the driving gas is utilized to control the injection of the reaction reagent, the reaction is actually injected into the reagent groove 5, different actual batch injection or common injection is realized, so that the quantitative injection of different reaction reagents is realized, the operation difficulty of operators is effectively simplified, and the structure of the reaction unit is simplified due to the matching of the gas path control plate 2 and the reaction chip; furthermore, the heating plate 1 is arranged to heat the reflecting chip, so that the invention has the function of reverse transcription, and has the characteristics of simple structure, simple operation, small occupied area, strong adaptability and the like.
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 disclosed herein fall within the scope and disclosure of the present invention.
Claims (10)
1. A cell sample preparation device is characterized by comprising a rack, wherein a reaction unit is arranged on the rack and comprises a gas circuit control board and a heating plate, and a reaction chip is clamped between the gas circuit control board and the heating plate;
at least two mutually independent driving air channel channels are arranged in the air channel control plate;
the reaction chip comprises a carrying platform and a cell reaction plate which are arranged in an attaching mode, wherein reagent grooves with the same number as that of the driving gas path channels are formed in the surface of one side, attached to the gas path control plate, of the cell reaction plate, and each driving gas path channel is independently communicated with one reagent groove;
a reaction cavity is formed in the surface of one side, attached to the carrier, of the cell reaction plate, and the reagent grooves are independently connected with the reaction cavity;
and the reagent groove is internally injected with a reaction reagent in advance, and the driving gas is injected into the reagent groove through the driving gas path, so that the reaction reagent in the reagent groove is pressed into the reaction cavity.
2. The manufacturing apparatus according to claim 1, wherein a product tank and a waste liquid tank are further disposed on a surface of the cell reaction plate on one side of the reagent tank, at least two air pumping channels are further disposed in the air path control plate, and the product tank and the waste liquid tank are respectively and independently connected to the air pumping channels;
preferably, the product tank and the waste liquid tank are respectively and independently connected with the reaction cavity;
preferably, the product tank and the waste liquid tank are both connected with an air suction electromagnetic valve through the air suction channel.
3. The manufacturing apparatus according to claim 1 or 2, wherein the reagent tanks are each connected to a gas injection solenoid valve through the driving gas passage;
preferably, the air exhaust solenoid valve and the air injection solenoid valve are both arranged on the same side surface of the air path control plate in a centralized manner.
4. The manufacturing apparatus according to any one of claims 1 to 3, wherein a buffer tank is provided on a surface of the cell reaction plate on one side of the reagent tank, the reagent tank and the reaction chamber are independently communicated with the buffer tank, and the reagent tank, the buffer tank and the reaction chamber are connected in sequence along a flow direction of the reaction reagent;
preferably, a silica gel pad is arranged between the cell reaction plate and the air channel control plate, and a hole corresponding to the outlet position of the driving air channel on the air channel control plate is formed in the silica gel pad.
5. The manufacturing apparatus as claimed in any one of claims 1 to 4, wherein the gas circuit control plate comprises a gas circuit upper plate and a gas circuit lower plate which are stacked;
preferably, the binding surface of the air path lower plate and the air path upper plate is provided with at least one air path groove, the air path upper plate is bound with the air path lower plate, and the air path groove is closed and sealed to form the driving air path channel and the air exhaust channel.
6. The manufacturing apparatus of any one of claims 1 to 5, wherein at least one independent reagent flow channel is further formed on a side surface of the cell reaction plate, which is attached to the carrier, and the reagent flow channel forms a reagent flow channel after the carrier and the cell reaction plate are attached and sealed;
preferably, the reagent reservoirs are independently connected to the buffer reservoirs through the reagent flow channels, respectively;
preferably, the buffer tanks are independently connected to the reaction chamber through the reagent flow channel;
preferably, the product tank and the waste tank are both independently connected to the reaction chamber through the reagent flow channel.
7. The preparation device according to any one of claims 1 to 7, wherein a control unit is arranged at the bottom of the reaction unit, the control unit is respectively and independently electrically connected with the heating plate, the gas injection solenoid valve and the gas extraction solenoid valve, and the control unit respectively and independently controls the activation of the heating plate, the activation of the gas injection solenoid valve and the activation of the gas extraction solenoid valve.
8. The preparation device according to claims 3-7, wherein the gas injection solenoid valve and the gas extraction solenoid valve are both connected with a gas pump assembly, and the gas pump assembly is used for providing the pressure of gas in the gas injection solenoid valve and the gas extraction solenoid valve;
preferably, the air pump assembly is located below the reaction unit;
preferably, the air pump assembly is arranged side by side with the control unit.
9. A method for preparing a cell sample using the cell sample preparation device according to any one of claims 1 to 8, said method comprising:
injecting a reaction reagent into the reagent cavity, placing the reaction chip between the air circuit control plate and the heating plate, injecting a driving gas into the reagent groove through the driving air circuit channel, pressing the reaction reagent in the reagent groove into the reaction cavity for cell reaction, and starting the heating plate to heat the reaction cavity for reverse transcription to prepare the cell sample.
10. The preparation method according to claim 9, wherein the preparation method specifically comprises the following steps:
after injecting a reaction reagent into the reagent cavity, placing the reaction chip between the gas path control plate and the heating plate, controlling the gas injection electromagnetic valve to be started by the control unit, injecting gas into the reagent cavity to press the reaction reagent into the buffer cavity, controlling the gas extraction electromagnetic valve of the waste liquid tank to be started by the control unit, and sucking the reaction reagent in the buffer cavity into the reaction cavity by the gas extraction of the waste liquid tank to perform cell reaction;
(II) repeating the operation of the step (I) at least once, wherein the control unit controls the heating plate to heat and reversely transcribe the reaction cavity to prepare the cell sample;
and (III) the control unit controls the starting of an air exhaust electromagnetic valve of the product tank to suck the cell sample in the reaction cavity into the product tank.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110535870.9A CN115353972A (en) | 2021-05-17 | 2021-05-17 | Cell sample preparation device and preparation method thereof |
| PCT/CN2021/135159 WO2022117053A1 (en) | 2020-12-02 | 2021-12-02 | Reagent exchange methods, devices, and systems |
| CN202180092485.XA CN116867889A (en) | 2020-12-02 | 2021-12-02 | Reagent exchange method, device and system |
| 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 |
|---|---|---|---|
| CN202110535870.9A CN115353972A (en) | 2021-05-17 | 2021-05-17 | Cell sample preparation device and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115353972A true CN115353972A (en) | 2022-11-18 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110535870.9A Pending CN115353972A (en) | 2020-12-02 | 2021-05-17 | Cell sample preparation device and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115353972A (en) |
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2021
- 2021-05-17 CN CN202110535870.9A patent/CN115353972A/en active Pending
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