CN112191288B - Unicellular separator based on unicellular sequencing technology - Google Patents
Unicellular separator based on unicellular sequencing technology Download PDFInfo
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- CN112191288B CN112191288B CN202011081514.6A CN202011081514A CN112191288B CN 112191288 B CN112191288 B CN 112191288B CN 202011081514 A CN202011081514 A CN 202011081514A CN 112191288 B CN112191288 B CN 112191288B
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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
- B01L3/502707—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 characterised by the manufacture of the container or its components
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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
- B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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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
- B01L3/502769—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 characterised by multiphase flow arrangements
- B01L3/502784—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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
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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
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Abstract
The invention discloses a single cell separation device based on a single cell sequencing technology, which comprises a machine body, wherein a separation cavity is arranged in the machine body, a confluence base is fixedly arranged between the left wall and the right wall of the lower side of the separation cavity, a microfluidic column is fixedly arranged on the upper end surface of the confluence base, microfluidic channel pipes are fixedly arranged in a circumferential array on the outer circumferential surface of the microfluidic column, a dripping inlet with an upward opening is arranged at the position of the center of a circle in the microfluidic column, each microfluidic channel pipe is internally communicated with a microfluidic channel with an outward opening, and an annular confluence groove with an upward opening is formed in the upper end surface of the confluence base and is connected with the outer side of the microfluidic channel pipe; the invention can realize the separation of single cells by using the micro-channel plate, and the micro-channel plate is deformed by applying micro pressure, thereby controlling the flowing tendency of the internal liquid of the micro-channel, being beneficial to the flowing of cell micro-flow liquid, and simultaneously being capable of automatically adding a fluorescent labeling reagent in the process of dripping the cell reagent, and being convenient for identifying cells.
Description
Technical Field
The invention relates to the technical field of single cell sequencing, in particular to a single cell separation device based on a single cell sequencing technology.
Background
Single cell transcriptome sequencing refers to a new technique for high throughput sequencing and analysis of RNA at the single cell level. Different from the results (only the average expression level of a large number of cells) obtained by sequencing conventional tissues or cell groups, single sequencing can deeply excavate specific information, the most important step of single cell sequencing is the separation of single cells, the single cell separation technology comprises the steps of micromanipulation separation, laser microdissection, flow cytometry, microfluidic technology and the like, the microfluidic technology is a cell separation technology with greater advantages, and the separation of the single cells can be realized by utilizing gravity centrifugation, hydrodynamics, electric field force and the like.
Disclosure of Invention
The invention aims to solve the technical problem of providing a single cell separation device based on a single cell sequencing technology, and solves the problems of liquid flow direction guide and inconvenience in adding a fluorescence labeling reagent of the microfluidic single cell separation device.
The invention is realized by the following technical scheme.
The invention relates to a single cell separation device based on a single cell sequencing technology, which comprises a machine body, wherein a separation cavity is arranged in the machine body, a confluence base is fixedly arranged between the left wall and the right wall of the lower side of the separation cavity, a microfluidic column is fixedly arranged on the upper end surface of the confluence base, microfluidic channel pipes are fixedly arranged on the circumferential array of the outer circumferential surface of the microfluidic column, a dripping port with an upward opening is arranged at the position of the center of a circle in the microfluidic column, a microfluidic channel with an outward opening is communicated in each microfluidic channel pipe, an annular confluence groove with an upward opening is arranged at the joint of the upper end surface of the confluence base and the outer side of the microfluidic channel pipe, a liquid outlet pipeline with a downward opening is arranged in the confluence base and under the dripping port, a confluence channel is communicated between the lower side of the confluence base and the circumferential wall of the upper side of the liquid outlet pipeline, and a control valve is fixedly arranged on the lower end surface of the confluence base and under the liquid outlet pipeline, thereby the control valve can control the whereabouts of the switching control liquid of drain pipe downside, thereby the organism is internal and communicate the separating chamber downside is equipped with about open-ended reagent board chamber, it is equipped with the reagent board to place on the reagent board chamber lower wall, evenly distributed is equipped with the ascending liquid groove of connecing of opening in the reagent board up end about, the fixed spliced pole that is equipped with of separating chamber upper wall, the fixed pressure disk that is equipped with of terminal surface under the spliced pole, terminal surface circumference array is equipped with the pressure column chamber that the opening is decurrent under the pressure disk, the gliding pressure column that is equipped with from top to bottom in the pressure column chamber, terminal surface offsets with miniflow pipe up end under the pressure column, make miniflow pipe produce the deformation that removes through pressure when the pressure column moves down to control the liquid flow in the equidirectional miniflow way, be equipped with the lift chamber in the spliced pole, the gliding lifter plate that is equipped with from top to bottom in the lift chamber, the fixed drip needle that is equipped with that runs through from top to bottom in the lifter plate, the fixed mixing block that is equipped with in drip needle middle part, be equipped with in the mixing block and switch the rotation chamber, it is equipped with mixed rotary drum to switch the rotation intracavity, it is equipped with the ascending connector of opening to mix rotary drum upside intercommunication, in the drip needle and mix a upside intercommunication switch the rotation chamber and be equipped with the ascending feed liquor passageway of opening, in the drip needle and be in mix a downside intercommunication switch the rotation chamber and be equipped with the decurrent drain passage of opening, intercommunication in the mixing block switches the upper right side of rotation chamber and is equipped with the interpolation passageway that the opening was right, in the spliced pole and go up and down the chamber right side with be equipped with the fluorescence reagent chamber between the rotary drum chamber, fluorescence reagent chamber downside left wall with the intercommunication is equipped with the connecting pipe between the interpolation passageway right side.
Preferably, just be in the organism reagent board chamber downside intercommunication is equipped with the translation chamber, the translation chamber is equipped with interior energy horizontal slip's slurcam spring, it accomodates the chamber to be equipped with the ascending slurcam of opening in the slurcam spring, the slurcam is accomodate the intracavity and is equipped with gliding slurcam from top to bottom, the slurcam can extend to reagent board intracavity and with reagent board left end face offsets, under the slurcam terminal surface with the slurcam accomodates fixedly connected with slurcam spring between the chamber lower wall, the slurcam under the terminal surface with the slurcam accomodates the chamber lower wall and fixes the electro-magnet that is equipped with respectively and can inhale mutually by opposite polarities, it is equipped with the translation screw rod to rotate between the wall about the translation chamber.
Preferably, a translation motor is fixedly arranged in the right wall of the translation cavity, and the right end of the translation screw is in power connection with the left end face of the translation motor.
Preferably, in the pressure dish and communicate all jointly pressure column chamber upside is equipped with the rotation chamber, be equipped with annular rotary drum chamber in the spliced pole, the rotary drum intracavity rotates and is equipped with the rotary drum, the rotary drum lower extreme extends to rotate intracavity and outer disc fixed pressure switching ring that is equipped with, pressure column upper end extends to rotate intracavity and the fixed board that resets of up end, reset under the board terminal surface with rotate between the chamber lower wall and around in pressure column outside fixedly connected with reset spring, the fixed lower briquetting that is equipped with down the terminal surface ability with the up end of reset plate offsets under the pressure switching ring left side terminal surface.
Preferably, the inner circular wall at the left side of the rotary cylinder cavity is communicated with a rotary gear cavity, a rotary driving shaft is rotatably arranged between the upper wall and the lower wall of the rotary gear cavity, a rotating gear meshed with the inner circular surface on the upper side of the rotary drum is fixedly arranged on the outer circular surface of the rotating driving shaft, a driving device is fixedly arranged in the upper wall of the rotating gear cavity, the upper end of the rotating driving shaft is in power connection with the lower end surface of the driving device, a lifting screw rod is rotatably arranged on the upper end surface of the pressure disc and on the left side of the dropping needle, a lifting driven gear is fixedly arranged on the outer circular surface of the upper end of the lifting screw rod, a lifting gear cavity with a right opening is arranged in the connecting column and at the upper side of the rotating gear cavity, a lifting driving shaft is rotatably arranged between the upper wall and the lower wall of the lifting gear cavity, and a lifting driving gear meshed with the lifting driven gear is fixedly arranged on the outer circular surface of the lifting driving shaft, and the lower end of the lifting driving shaft is connected with the upper end surface of the driving device through a cable.
Preferably, the outer round surface of the mixing drum is internally provided with arc-shaped rotating teeth fixedly arranged at the rear side of the mixing cavity, the left side of the switching rotating cavity is communicated with a driving gear cavity, a driving gear shaft is rotatably arranged between the front wall and the rear wall of the driving gear cavity, a driving gear meshed with the rotating teeth is fixedly arranged on the outer round surface of the driving gear shaft, a micro motor is fixedly arranged in the rear wall of the driving gear cavity, and the rear end of the driving gear shaft is in power connection with the front end face of the micro motor.
The invention has the beneficial effects that: the invention can realize the separation of single cells by using the micro-channel plate, and the micro-channel tube deforms by applying micro pressure, thereby controlling the flowing tendency of the internal liquid of the micro-channel, being beneficial to the flowing of cell micro-flow liquid, and simultaneously being capable of automatically adding a fluorescent labeling reagent in the process of dripping the cell reagent, and being convenient for identifying cells.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the invention, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a schematic structural diagram of an embodiment of the present invention;
FIG. 2 is an enlarged schematic view of the embodiment of the present invention at A in FIG. 1;
FIG. 3 is a schematic structural diagram of B-B in FIG. 2 according to an embodiment of the present invention;
FIG. 4 is an enlarged schematic view of the embodiment of the present invention at C in FIG. 1;
FIG. 5 is an enlarged schematic view of the embodiment of the present invention at D in FIG. 1;
FIG. 6 is an enlarged schematic view of the embodiment of the present invention at E in FIG. 1.
Detailed Description
The invention will now be described in detail with reference to fig. 1-6, wherein for ease of description the orientations described hereinafter are now defined as follows: the up, down, left, right, and front-back directions described below correspond to the up, down, left, right, and front-back directions in the projection relationship of fig. 1 itself.
The unicell separation device based on the unicell sequencing technology, which is described in conjunction with the accompanying drawings 1-6, includes a machine body 11, a separation chamber 12 is arranged in the machine body 11, a confluence base 14 is fixedly arranged between the left and right walls of the lower side of the separation chamber 12, a microfluidic column 76 is fixedly arranged on the upper end surface of the confluence base 14, microfluidic channel pipes 31 are fixedly arranged on the circumferential array of the outer circumferential surface of the microfluidic column 76, a dripping port 33 with an upward opening is arranged at the inner center of the microfluidic column 76, a microfluidic channel 32 with an outward opening is communicated in each microfluidic channel pipe 31, an annular confluence groove 30 with an upward opening is arranged at the joint of the upper end surface of the confluence base 14 and the outer side of the microfluidic channel pipe 31, a liquid outlet pipe 28 with a downward opening is arranged in the confluence base 14 and under the dripping port 33, a confluence channel 29 is communicated between the lower side of the confluence groove 30 and the circumferential wall of the upper side of the liquid outlet pipe 28, the lower end face of the confluence base 14 is fixedly provided with a control valve 24 under the liquid outlet pipeline 28, the control valve 24 can control the opening and closing of the lower side of the liquid outlet pipeline 28 to control the falling of liquid, the machine body 11 is communicated with a reagent plate cavity 13 with a left opening and a right opening, which is arranged at the lower side of the separation cavity 12, a reagent plate 23 is arranged on the lower wall of the reagent plate cavity 13, liquid receiving grooves 25 with upward openings are uniformly distributed in the upper end face of the reagent plate 23 from left to right, a connecting column 15 is fixedly arranged on the upper wall of the separation cavity 12, a pressure disc 16 is fixedly arranged on the lower end face of the connecting column 15, a pressure column cavity 71 with a downward opening is arranged on the circumferential array of the lower end face of the pressure disc 16, a pressure column 54 capable of sliding up and down is arranged in the pressure column cavity 71, the lower end face of the pressure column 54 is abutted against the upper end face of the micro-flow channel pipe 31, and the micro-channel pipe 31 is deformed by pressure when the pressure column 54 moves downwards, thereby controlling the liquid flow in the equidirectional microchannel 32 of equidirectional, be equipped with lift chamber 70 in the spliced pole 15, the gliding lifter plate 35 that is equipped with about in the lift chamber 70, be equipped with the fixed drip needle 36 that runs through from top to bottom in the lifter plate 35, drip needle 36 middle part is fixed and is equipped with mixing block 39, be equipped with in the mixing block 39 and switch and rotate chamber 41, switch and rotate the intracavity 41 internal rotation and be equipped with mixing drum 42, mixing drum 42 upside intercommunication is equipped with the ascending connector 44 of opening, in the drip needle 36 and be in mixing block 39 upside intercommunication switch and rotate chamber 41 and be equipped with the ascending inlet channel 37 of opening, in the drip needle 36 and be in mixing block 39 downside intercommunication switch and rotate chamber 41 and be equipped with the decurrent outlet channel 38 of opening, the intercommunication in the mixing block 39 switching is rotated the upper right side of chamber 41 and is equipped with the right interpolation passageway 50 of opening, a fluorescent reagent cavity 19 is arranged between the right side of the lifting cavity 70 and the rotary drum cavity 20 in the connecting column 15, and a connecting pipe 40 is communicated between the left wall of the lower side of the fluorescent reagent cavity 19 and the right side of the adding channel 50.
Beneficially, translation chamber 22 is arranged in the machine body 11 and communicated with the lower side of the reagent plate chamber 13, a pushing plate spring 73 capable of sliding left and right is arranged in the translation chamber 22, a pushing plate accommodating chamber 75 with an upward opening is arranged in the pushing plate spring 73, a pushing plate 57 capable of sliding up and down is arranged in the pushing plate accommodating chamber 75, the pushing plate 57 can extend into the reagent plate chamber 13 and abut against the left end face of the reagent plate 23, a pushing plate spring 73 is fixedly connected between the lower end face of the pushing plate 57 and the lower wall of the pushing plate accommodating chamber 75, electromagnets 56 capable of absorbing in opposite directions are fixedly arranged on the lower end face of the pushing plate 57 and the lower wall of the pushing plate accommodating chamber 75 respectively, and translation screw rods 26 are arranged between the left wall and the right wall of the translation chamber 22 in a rotating manner.
Advantageously, a translation motor 27 is fixedly arranged in the right wall of the translation cavity 22, and the right end of the translation screw 26 is in power connection with the left end face of the translation motor 27.
Beneficially, a rotating cavity 17 is formed in the pressure disc 16 and is communicated with the upper sides of all the pressure column cavities 71 together, an annular drum cavity 20 is formed in the connecting column 15, a drum 21 is rotatably formed in the drum cavity 20, the lower end of the drum 21 extends into the rotating cavity 17, a pressure switching ring 18 is fixedly arranged on the outer circular surface of the drum, the upper end of the pressure column 54 extends into the rotating cavity 17, a reset plate 53 is fixedly arranged on the upper end surface of the pressure column 54, a reset spring 55 is fixedly connected between the lower end surface of the reset plate 53 and the lower wall of the rotating cavity 17 and surrounds the outer side of the pressure column 54, and a lower press block 52, the lower end surface of which can abut against the upper end surface of the reset plate 53, is fixedly arranged on the lower end surface of the left side of the pressure switching ring 18.
Advantageously, a rotating gear chamber 64 is communicated with the inner wall of the left side of the drum chamber 20, a rotating driving shaft 72 is rotatably arranged between the upper wall and the lower wall of the rotating gear chamber 64, a rotating gear 66 engaged with the inner wall of the upper side of the drum 21 is fixedly arranged on the outer circumferential surface of the rotating driving shaft 72, a driving device 63 is fixedly arranged on the upper wall of the rotating gear chamber 64, the upper end of the rotating driving shaft 72 is connected with the lower end surface of the driving device 63 in a power manner, a lifting screw 62 is rotatably arranged on the upper end surface of the pressure plate 16 and on the left side of the drip needle 36, a lifting driven gear 61 is fixedly arranged on the outer circumferential surface of the upper end of the lifting screw 62, a lifting gear chamber 58 with a right opening is arranged in the connecting column 15 and on the upper side of the rotating gear chamber 64, a lifting driving shaft 59 is rotatably arranged between the upper wall and the lower wall of the lifting gear chamber 58, a lifting driving gear 60 engaged with the lifting driven gear 61 is fixedly arranged on the outer circumferential surface of the lifting driving shaft 59, the lower end of the lifting driving shaft 59 is connected with the upper end surface of the driving device 63 through a cable.
Beneficially, an arc-shaped rotating tooth 45 is fixedly arranged in the outer circumferential surface of the mixing drum 42 and at the rear side of the mixing chamber 43, a driving gear chamber 47 is communicated with the left side of the switching rotating chamber 41, a driving gear shaft 49 is rotatably arranged between the front wall and the rear wall of the driving gear chamber 47, a driving gear 48 meshed with the rotating tooth 45 is fixedly arranged on the outer circumferential surface of the driving gear shaft 49, a micro motor 51 is fixedly arranged in the rear wall of the driving gear chamber 47, and the rear end of the driving gear shaft 49 is dynamically connected to the front end surface of the micro motor 51.
In the initial state, the lifting plate 35 is located at the upper limit position, the dropping needle 36 is located at the upper limit position, all the pressure columns 54 are located at the upper limit position, the connecting port 44 rotates to be communicated with the liquid inlet channel 37, the pushing plate 57 is located at the upper limit position, the right end face of the pushing plate is abutted against the left end face of the reagent plate 23, and the reset plate 53 is located at the left limit position.
When the device works, a reagent with cells flows into the mixing cavity 43 through the liquid inlet channel 37 and the connecting port 44, then the driving device 63 is started, the lifting driving shaft 59 is driven to rotate and the lifting driving gear 60 is driven to rotate through power connection, the lifting driven gear 61 is driven to rotate and the lifting screw 62 is driven to rotate through gear meshing, the lifting plate 35 is driven to move downwards through threaded connection, the dropping needle 36 is driven to move downwards until the lower end extends into the dropping opening 33, then the micro motor 51 is started, the driving gear shaft 49 is driven to rotate and the driving gear 48 is driven to rotate through power connection, the rotating teeth 45 is driven to rotate and the mixing drum 42 is driven to rotate through gear meshing, then the connecting port 44 is rotated to be communicated with the adding channel 50, and then a fluorescence labeling reagent in the fluorescence labeling reagent cavity 19 enters the mixing cavity 43 through the connecting pipe 40 and the adding channel 50 to be mixed with the cell reagent, then the mixing drum 42 continues to rotate to connect the connection port 44 with the liquid outlet channel 38, and then the mixed reagent in the mixing chamber 43 flows out downwards through the connection port 44 and the liquid outlet channel 38 to the drop port 33, at this time, the driving device 63 drives the rotating driving shaft 72 to rotate through power connection and drives the rotating gear 66 to rotate, further drives the drum 21 to rotate through gear engagement, further drives the pressure switching ring 18 to rotate and drives the lower pressing block 52 to rotate, thereby rotating the lower pressing block 52 to the upper end surface of one of the returning plates 53, further pressing the returning plate 53 downwards and compressing the returning spring 55, further driving the pressure column 54 to move downwards, further causing the micro flow channel tube 31 to deform and control and guide the micro fluid in the corresponding micro flow channel 32 to move away from the drop port 33, thereby flowing into the confluence groove 30 and flowing into the liquid outlet channel 28 through the confluence channel 29, the liquid in the liquid outlet pipeline 28 is controlled to flow out through the control valve 24, the translation motor 27 is started, the translation screw rod 26 is driven through power connection, the translation slide block 74 is driven to move rightwards through threaded connection, the push plate 57 is driven to move rightwards and the reagent plate 23 is driven to move rightwards, each liquid receiving groove 25 is filled with cell unicellular liquid, the reagent plate 23 is driven to move rightwards to the right side of the machine body 11 to be taken out conveniently, then the translation motor 27 is started reversely, the push plate 57 is driven to move leftwards through corresponding transmission to reset, the electromagnet 56 is electrified, the push plate 57 is driven to move downwards through magnetic force opposite attraction and pushes the plate spring 73, and then the next reagent plate 23 is placed onto the lower wall of the reagent plate cavity 13 from the left side of the reagent plate cavity 13 to be taken out for the next reagent plate.
The above embodiments are merely illustrative of the technical ideas and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.
Claims (5)
1. A unicellular separator based on unicellular sequencing technology, includes the organism, its characterized in that: a separation cavity is arranged in the machine body, a confluence base is fixedly arranged between the left wall and the right wall of the lower side of the separation cavity, a micro flow column is fixedly arranged on the upper end face of the confluence base, a micro flow channel pipe is fixedly arranged on the circumferential array of the outer circumferential surface of the micro flow column, a dripping port with an upward opening is arranged at the position of the center of a circle in the micro flow column, a micro flow channel with an outward opening is arranged in each micro flow channel pipe in a communicated manner, an annular confluence groove with an upward opening is arranged at the joint of the upper end face of the confluence base and the outer side of the micro flow channel pipe, a liquid outlet pipeline with a downward opening is arranged in the confluence base and under the dripping port, a confluence channel is arranged between the lower side of the confluence groove and the upper side of the circumferential wall of the liquid outlet pipeline in a communicated manner, a control valve is fixedly arranged on the lower end face of the confluence base and under the liquid outlet pipeline, and can control the opening and closing of the lower side of the liquid outlet pipeline so as to control the falling of liquid, open-ended reagent board chamber about just communicateing the separation chamber downside is equipped with in the organism, it is equipped with the reagent board to place on the reagent board chamber lower wall, evenly distributed is equipped with the ascending cistern that connects of opening about in the reagent board up end, the fixed spliced pole that is equipped with of separation chamber upper wall, the fixed pressure disk that is equipped with of terminal surface under the spliced pole, terminal surface circumference array is equipped with the pressure column chamber that the opening is decurrent under the pressure disk, be equipped with gliding pressure column from top to bottom in the pressure column chamber, terminal surface offsets with microchannel pipe up end under the pressure column, make microchannel pipe produce the deformation that removes through pressure when the pressure column moves down to the liquid flow in the control equidirectional microchannel, be equipped with the lift chamber in the spliced pole, gliding lifter plate is equipped with from top to bottom in the lift chamber, the lifter plate is equipped with the fixed drip needle that runs through from top to bottom in, the mixing block is fixedly arranged in the middle of the liquid dropping needle, a switching rotating cavity is arranged in the mixing block, a mixing rotary drum is rotatably arranged in the switching rotating cavity, a connecting port with an upward opening is formed in the upper side of the mixing rotary drum in a communicated manner, a liquid inlet channel with an upward opening is formed in the mixing block in a communicated manner, a liquid outlet channel with a downward opening is formed in the switching rotating cavity in the liquid dropping needle and in a communicated manner at the lower side of the mixing block, an adding channel with a rightward opening is formed in the mixing block in a communicated manner at the upper right side of the switching rotating cavity, a rotating cavity is formed in the pressure disc and is communicated with all the upper sides of the pressure column cavities together, an annular rotary drum cavity is arranged in the connecting column, a rotary drum is rotatably arranged in the rotary drum cavity, the lower end of the rotary drum extends into the rotary cavity, a pressure switching ring is fixedly arranged on the outer circular surface of the rotary cavity, and a reset plate is fixedly arranged on the upper end surface of the rotary cavity, reset under the board terminal surface with rotate between the chamber lower wall and around in pressure column outside fixedly connected with reset spring, under the pressure switching ring left side terminal surface fixed be equipped with down the terminal surface can with the lower briquetting that resets the counterbalance of board up end, just in the spliced pole go up and down the chamber right side with be equipped with the fluorescence reagent chamber between the rotary drum chamber, fluorescence reagent chamber downside left wall with the intercommunication is equipped with the connecting pipe between the interpolation passageway right side.
2. A single-cell separation device based on single-cell sequencing technology as claimed in claim 1, characterized in that: the internal just is in reagent board chamber downside intercommunication is equipped with the translation chamber, the translation intracavity is equipped with the slurcam spring that can the horizontal slip, it accomodates the chamber to be equipped with the ascending slurcam of opening in the slurcam spring, the slurcam is accomodate the intracavity and is equipped with gliding slurcam from top to bottom, the slurcam can extend to reagent board intracavity and with reagent board left end face offsets, the slurcam lower terminal surface with the slurcam accomodates fixedly connected with slurcam spring between the chamber lower wall, the slurcam lower terminal surface with the slurcam accomodates the chamber lower wall and is fixed respectively to be equipped with the electro-magnet that can opposite sex looks inhale, it is equipped with the translation screw rod to rotate between the wall about the translation chamber.
3. A single-cell separation device based on single-cell sequencing technology as claimed in claim 2, characterized in that: translation chamber right side wall internal fixation is equipped with the translation motor, translation screw rod right-hand member power connect in translation motor left end face.
4. A single-cell separation device based on single-cell sequencing technology as claimed in claim 1, characterized in that: the inner circular wall at the left side of the rotary cylinder cavity is communicated with a rotary gear cavity, a rotary driving shaft is rotatably arranged between the upper wall and the lower wall of the rotary gear cavity, a rotating gear meshed with the inner circular surface on the upper side of the rotary drum is fixedly arranged on the outer circular surface of the rotating driving shaft, a driving device is fixedly arranged in the upper wall of the rotating gear cavity, the upper end of the rotating driving shaft is in power connection with the lower end surface of the driving device, a lifting screw rod is rotatably arranged on the upper end surface of the pressure disc and on the left side of the dropping needle, a lifting driven gear is fixedly arranged on the outer circular surface of the upper end of the lifting screw rod, a lifting gear cavity with a right opening is arranged in the connecting column and at the upper side of the rotating gear cavity, a lifting driving shaft is rotatably arranged between the upper wall and the lower wall of the lifting gear cavity, and a lifting driving gear meshed with the lifting driven gear is fixedly arranged on the outer circular surface of the lifting driving shaft, and the lower end of the lifting driving shaft is connected with the upper end surface of the driving device through a cable.
5. A single-cell separation device based on single-cell sequencing technology as claimed in claim 1, characterized in that: the utility model discloses a mixing drum, including mixing drum, switching rotation chamber, driving gear axle excircle, mixing drum excircle in the face and at the fixed curved rotation tooth that is equipped with in mixing chamber rear side, switching rotation chamber left side intercommunication is equipped with the driving gear chamber, it is equipped with driving gear axle to rotate between the front and back wall in driving gear chamber, driving gear axle excircle fixed be equipped with the driving gear of rotation tooth meshing, driving gear chamber back wall internal fixation is equipped with the micromotor, driving gear axle rear end power connect in terminal surface before the micromotor.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011081514.6A CN112191288B (en) | 2020-10-12 | 2020-10-12 | Unicellular separator based on unicellular sequencing technology |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011081514.6A CN112191288B (en) | 2020-10-12 | 2020-10-12 | Unicellular separator based on unicellular sequencing technology |
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| CN112191288A CN112191288A (en) | 2021-01-08 |
| CN112191288B true CN112191288B (en) | 2021-12-24 |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005108963A1 (en) * | 2004-05-06 | 2005-11-17 | Nanyang Technological University | Microfluidic cell sorter system |
| WO2010115167A2 (en) * | 2009-04-03 | 2010-10-07 | The Regents Of The University Of California | Methods and devices for sorting cells and other biological particulates |
| JP5806236B2 (en) * | 2010-01-13 | 2015-11-10 | セブンス センス バイオシステムズ，インコーポレーテッド | Rapid delivery and / or collection of fluids |
| EP3281703B1 (en) * | 2013-04-15 | 2019-08-21 | Becton, Dickinson and Company | Biological fluid collection device and biological fluid separation system |
| CN104774747B (en) * | 2015-04-14 | 2017-03-01 | 浙江大学 | Microfluidic droplet chip apparatus for cell migration assay experiment and method |
| KR101986016B1 (en) * | 2017-09-12 | 2019-06-04 | 경희대학교 산학협력단 | Trapping tool single cells and its manufacturing method |
| CN110743636A (en) * | 2019-10-27 | 2020-02-04 | 苏州济研生物医药科技有限公司 | Droplet generation chip, preparation method thereof and application thereof in single cell sequencing |
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Effective date of registration: 20211201 Address after: 650000 Cell Industry Cluster Innovation Park, No. 101, Keyuan Road, high tech Zone, Kunming, Yunnan Applicant after: Yunnan Cell Engineering Center Co.,Ltd. Address before: 510000 a910, No.3, Juquan Road, Huangpu District, Guangzhou City, Guangdong Province Applicant before: Guangzhou isoda Biomedical Technology Co.,Ltd. |
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