CN217699228U - A micro-fluidic chip for unicellular automation is built storehouse - Google Patents
A micro-fluidic chip for unicellular automation is built storehouse Download PDFInfo
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- CN217699228U CN217699228U CN202220385354.2U CN202220385354U CN217699228U CN 217699228 U CN217699228 U CN 217699228U CN 202220385354 U CN202220385354 U CN 202220385354U CN 217699228 U CN217699228 U CN 217699228U
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Abstract
The utility model provides a micro-fluidic chip for single cell automatic warehouse building, which comprises a cover plate, a clamping plate and a chip plate; the upper surface of the clamping plate is attached to the cover plate, and the lower surface of the clamping plate is attached to the chip plate; one end of the cover plate is at least provided with 4 agent inlets; one end of the clamping plate is provided with a through hole corresponding to each agent inlet, the agent inlets are respectively communicated with one end of the through holes, the other end of each through hole is correspondingly communicated with one end of the flow guide groove, the other end of the flow guide groove is provided with a runner groove communicated with the flow guide groove, and the runner groove penetrates through the upper surface and the lower surface of the clamping plate; the chip board is correspondingly provided with a channel with the same specification as each runner groove, and the bottom wall of the channel is provided with a plurality of micropores with the hole spacing smaller than 5 mu m. The utility model not only reduces the production cost, but also can be perfectly matched with an automatic warehouse building platform, thereby greatly saving the manpower and improving the working efficiency; meanwhile, the multi-channel design increases the detection flux, each channel is independently loaded, and samples are not influenced mutually.
Description
Technical Field
The utility model relates to a cell and molecular biology detection area particularly, relate to a micro-fluidic chip that is used for automatic storehouse of building of unicellular.
Background
With the stability of the method and the continuous progress of the experimental platform, sequencing of single-cell transcriptome has become one of the most popular scientific technologies at present. From the initial manual operation method to the current high-throughput large-scale platform with great heat, researchers can utilize the technologies of all layers to deeply excavate the heterogeneity of cells according to different research purposes. The microplate technology platform in the single cell sequencing technology can capture hundreds to thousands of single cells and print bar codes on the single cells by utilizing a large number of micropores, and then genome and proteome information is analyzed. However, large scale operation is labor and time consuming. In addition, at present, two commercialized single cell technology platforms (10X Genomics and BD Rhapsody) have been widely accepted by domestic scientists, but the related high-precision technology is still mastered by foreign enterprises, a local service company has no very large autonomy, many aspects are limited by technical monopoly, one point which can be embodied most directly is probably the price aspect, and the first impression of many users on single cells is that the price is high, and the cost burden is heavy.
Therefore, how to provide a microfluidic chip for single cell automated library establishment with low production cost, high flux and good detection effect is a technical problem to be solved urgently by domestic research and development personnel at present.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a micro-fluidic chip that is used for unicellular automation to build storehouse to improve above-mentioned technical problem.
In order to achieve the purpose, the following technical scheme is adopted:
the utility model provides a micro-fluidic chip for the automatic single-cell warehouse building, which comprises a cover plate, a clamping plate and a chip plate;
the upper surface of the clamping plate can be selectively attached to the cover plate, and the lower surface of the clamping plate can be selectively attached to the upper surface of the chip plate;
one end of the cover plate is at least provided with 4 agent inlets;
one end of the clamping plate is provided with a through hole corresponding to each agent inlet, the agent inlets are respectively communicated with one end of the through holes, the other end of each through hole is correspondingly communicated with one end of the flow guide groove, the other end of the flow guide groove is provided with a runner groove communicated with the flow guide groove, and the runner groove penetrates through the upper surface and the lower surface of the clamping plate;
the upper surface of the chip plate is correspondingly provided with a plurality of channels with the same specification as the plurality of runner grooves on the clamping plate, and the bottom walls of the plurality of channels contain a plurality of micropores with the space between the plurality of micropores less than 5 mu m.
Furthermore, a chemical injection platform is arranged at the position, corresponding to the through hole, of the cover plate, the chemical injection platform protrudes out of the upper surface of the cover plate, and a chemical injection port communicated with the through hole is formed in the chemical injection platform.
Further, a gasket is arranged at the edge of the agent inlet.
Furthermore, the outer surface of the cover plate is correspondingly provided with a groove with the same specification as each runner groove.
Further, the length of the clamping plate is 120.2mm, the width of the clamping plate is 78.2mm, and the distance between the runner grooves is 18mm.
Furthermore, the width of the runner groove is 10mm-15mm, and the length of the runner groove is 80-100mm.
Further, the channels of the chip plate are tapered at the end of the through-hole.
Further, each channel of the chip plate contains at least 15 ten thousand microwells.
Further, the structure of each micropore is in a pyramid shape.
Further, the microfluidic chip also comprises a sealing plug matched with the agent inlet.
Compared with the prior art, the utility model discloses following beneficial effect has:
the micro-fluidic chip provided by the utility model can be produced in large scale by modern machining mode, so as to greatly reduce the production cost; and perfect matching with an automatic library building platform is realized, so that labor is greatly saved, and the efficiency is improved. And simultaneously, the utility model provides a micro-fluidic chip uses 4 passageway micropore designs, and every passageway is the appearance of going up alone, each other does not influence, and measuring channel's nimble change, micropore quantity can reach 60 ten thousand, and the measuring flux scope is wide.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention, and therefore should not be considered as limiting the scope, and that those skilled in the art can also obtain other related drawings based on these drawings without inventive efforts.
Fig. 1 is a schematic structural diagram of a cover plate according to an embodiment of the present invention;
fig. 2 is a schematic structural view of a splint according to an embodiment of the present invention;
fig. 3 is a schematic structural diagram of a chip board according to an embodiment of the present invention;
fig. 4 is a sectional view of a diversion trench provided in an embodiment of the present invention;
fig. 5 is a cross-sectional view of a micro-hole provided in an embodiment of the present invention;
fig. 6 is a cross-sectional view of a combination of a cover plate, a clamping plate, a chip plate and a sealing plug according to an embodiment of the present invention.
Reference numeral 1-cover plate; 2-a dosing table; 3-an agent inlet; 4-a groove; 5-clamping plates; 6-runner groove; 7-a through hole; 8-chip board; 9-channel; 10-micropores; 11-magnetic beads; 12-a diversion trench; 13-sealing plug.
Detailed Description
The technical solution of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate the position or positional relationship based on the position or positional relationship shown in the drawings, or the position or positional relationship which is usually placed when the product of the present invention is used, and are only for convenience of description and simplification of the description, but do not indicate or imply that the device or element referred to must have a specific position, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.
Examples
The microfluidic chip for the single cell automatic library building according to fig. 1 to 3 comprises a cover plate 1, a clamping plate 5 and a chip plate 8, which are detachable independent components. Referring to fig. 6, in the working state, the upper surface of the clamping plate 5 is attached to the cover plate 1, and the lower surface of the clamping plate 5 is attached to the chip board 8.
When the three components are attached, the cover plate 1 is located at the uppermost end, which enables the microfluidic chip to be integrally formed into a sealed environment without interference from other factors during operation.
The clamping plate 5 is provided with a runner groove 6 which can form a completely closed liquid channel when being attached to the cover plate 1 and the chip plate 8.
The chip plate 8 is the place where the single cell trapping is done. When the magnetic beads 11 are introduced into the solution, the magnetic beads 11 can be bound by the chip plate 8 and fully contact with the solution added later until the single cell capture is completed.
Referring to fig. 3, the chip plate 8 is provided with a plurality of channels 9 at positions corresponding to the channel grooves 6, respectively, each channel 9 having the same size as the channel groove 6, and the chip channel 9 is tapered at the end of the through-hole 7. Referring to fig. 5, the channels 9 contain a plurality of micropores 10 with a pore spacing of less than 5 μm, each channel 9 contains at least 15 ten thousand micropores 10, and the micropores 10 are in the shape of rhombus cones.
The chip channel 9 is designed to be gradually enlarged in a conical shape at the end of the through hole 7, so as to ensure that liquid can be rapidly distributed in the liquid flow channel after entering the liquid flow channel.
Meanwhile, each channel 9 contains about 150000 rectangular pyramid linear arrays, the wells 10 are flat-bottomed, and the well spacing is <5um. Based on the principle of 'poisson distribution', the arrangement enables the sample loading cells and the magnetic beads 11 to naturally settle into the micropores 10, and ensures that each micropore 10 only falls into one magnetic bead 11 and one cell to form an independent reaction environment.
Referring to fig. 2, the length of the clamping plate 5 is set to 120.2mm, the width is set to 78.2mm, and the distance between the runner grooves 6 is 18mm; the width of the runner groove 6 is 10mm-15mm, and the length is 80-100mm. One end of the clamping plate 5 is provided with a through hole 7 corresponding to each agent inlet 3, and the agent inlets 3 are respectively communicated with one end of the through hole 7. Referring to fig. 4, the other end of each through hole 7 is correspondingly communicated with one end of a guide groove 12, the guide groove 12 is formed in the lower surface of the clamping plate 5, the other end of the guide groove 12 is provided with a runner groove 6 communicated therewith, and the runner groove 6 penetrates through the upper surface and the lower surface of the clamping plate 5.
The length and the width of the clamping plate 5, the length and the width of the runner groove 6 and the distance between the grooves are set according to an automatic workbench of a microfluidic chip, and can be perfectly matched with an automatic library building platform.
The guide groove 12 does not penetrate through the upper surface and the lower surface of the clamping plate 5, so the groove thickness of the guide groove 12 is smaller than that of the flow channel groove 6, one end of the guide groove 12 is communicated with the through hole 7, the through hole 7 needs to be matched with a pipetting gun head, and the size of the through hole 7 is far smaller than that of the flow channel groove 6.
The maximum flow of the diversion trench 12 is smaller than the maximum flow of the runner trench 6, and in the process of liquid inflow, the liquid is controlled to slowly enter the runner trench 6 at a constant speed, so that magnetic beads 11 caused by the fact that a large amount of liquid flows into the runner trench 6 in a short time are prevented from being washed away from the micropores 10.
Referring to fig. 1, a chemical feeding table 2 is arranged at a position of the cover plate 1 corresponding to the through hole 7, the chemical feeding table 2 protrudes out of the upper surface of the cover plate 1, a chemical feeding port 3 communicated with the through hole 7 is arranged on the chemical feeding table 2, and a gasket is arranged at the edge of the chemical feeding port 3. While the cover plate 1 is correspondingly provided with a groove 4 having the same size as each runner groove 6.
The agent inlet platform 2 protrudes from the upper surface of the cover plate 1, so that the agent inlet 3 on the agent inlet platform 2 is convenient for the insertion of the pipette tips.
The gasket is provided to improve the sealing properties of the pipette tip when inserted.
The cover plate 1 is correspondingly provided with the groove 4 with the same specification as each runner groove 6, so that when the three components are attached, the groove 4 on the cover plate 1, the runner groove 6 on the clamping plate 5 and the channel 9 on the chip plate 8 form a completely closed liquid runner, the reaction volume in the reaction tank can be reduced, and the reaction efficiency is improved; meanwhile, the height difference between the chip and the reagent outlet can be formed, and the liquid in the chip flow channel can be discharged only under the condition of applying pressure.
In addition, the micro-fluidic chip also comprises a sealing plug 13 matched with the agent inlet 3.
The sealing plug 13 adapted to the inlet 3 is provided to ensure the sealing property of the liquid channel after the magnetic beads 11 and the solution to be measured are added.
The utility model discloses the apron that includes 1, splint 5, chip board 8, three subassembly is laminated each other, wherein splint 5 is located between chip board 8 and the apron 1, splint 5's upper surface corresponds the lower surface of apron 1, splint 5's lower surface corresponds the upper surface of chip board 8, runner groove 6 runs through splint 5's upper surface and lower surface, consequently, after three subassembly laminating, runner groove 6's cell wall and apron 1's lower surface, enclose between chip board 8's the upper surface and close and form complete liquid runner, so that let in the back with liquid from income agent mouth 3, liquid flows runner groove 6.
At least four liquid flow channels are arranged on each micropore 10 chip, so that the detection efficiency is improved, and samples are independently loaded through each flow channel, so that the samples among the liquid flow channels cannot be influenced mutually, and the detection accuracy is improved.
When the microfluidic chip is used for single cell capture, the operation method comprises the following steps:
before the experiment, fix apron 1, splint 5 and chip board 8 through the fixed technology of chemical bonding, ensure to laminate between the upper surface of splint 5 and the lower surface of apron 1, closely laminate between the upper surface of splint 5 lower surface and chip board 8 to realize the leakproofness of laminating department separately, avoid runner groove 6 to produce the weeping.
The experimental process comprises the following steps: before the magnetic beads 11 and the sample to be detected are added, ethanol is added from the reagent inlet 3 to remove air bubbles in the micropores 10, then the ethanol is washed away by PBS solution, and then single cell suspension is added, and the cells freely sink into the micropores 10 under the action of gravity and are attached to the magnetic beads 11. Then, more magnetic beads 11 than the number of wells 10 are added, so that one magnetic bead 11 is contained in each well 10. Then, a pressurized liquid is introduced into the inlet 3 to blow off the excess magnetic beads 11. Then cell lysis solution is introduced to release the nucleic acid of the cells in each micro-well 10, the specific sequence carried on the magnetic beads 11 captures the free nucleic acid sequence, different tag sequences on the magnetic beads 11 can distinguish each cell, finally, the magnetic beads 11 are sucked out from the micro-wells 10 by using a magnet, and the magnetic beads 11 sucked out from each liquid flow channel are collected, so that the capture of the cells is completed.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A micro-fluidic chip for single cell automatic library building is characterized by comprising a cover plate, a clamping plate and a chip plate;
the upper surface of the clamping plate can be selectively attached to the cover plate, and the lower surface of the clamping plate can be selectively attached to the upper surface of the chip plate;
one end of the cover plate is at least provided with 4 agent inlets;
one end of the clamping plate is provided with a through hole corresponding to each agent inlet, the agent inlets are respectively communicated with one end of the through holes, the other end of each through hole is correspondingly communicated with one end of a flow guide groove, the other end of each flow guide groove is provided with a flow channel groove communicated with the flow guide groove, and the flow channel grooves penetrate through the upper surface and the lower surface of the clamping plate;
the upper surface of the chip plate is correspondingly provided with a plurality of channels with the same specification as the plurality of runner grooves on the clamping plate, and the bottom walls of the channels are provided with micropores with a plurality of hole intervals smaller than 5 mu m.
2. The microfluidic chip according to claim 1, wherein the cover plate is provided with an agent inlet platform corresponding to the through hole, the agent inlet platform protrudes out of the upper surface of the cover plate, and the agent inlet is located on the agent inlet platform.
3. The microfluidic chip according to claim 2, wherein the edge of the inlet is provided with a gasket.
4. The microfluidic chip according to claim 1, wherein the outer surface of the cover plate is correspondingly provided with a groove having the same size as each runner channel.
5. The microfluidic chip according to claim 4, wherein the length of the clamping plate is 120.2mm, the width of the clamping plate is 78.2mm, and the distance between the runner channels is 18mm.
6. The microfluidic chip according to claim 5, wherein the channel width is 10mm to 15mm, and the channel length is 80 mm to 100mm.
7. The microfluidic chip according to claim 1, wherein the channels of the chip plate are tapered at the end of the through-hole.
8. The microfluidic chip according to claim 7, wherein each channel of the chip plate comprises at least 15 ten thousand microwells.
9. The microfluidic chip according to claim 8, wherein the structure of the micro-pores is rhombus-shaped.
10. The microfluidic chip according to claim 1, further comprising a sealing plug adapted to the inlet.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202220385354.2U CN217699228U (en) | 2022-02-24 | 2022-02-24 | A micro-fluidic chip for unicellular automation is built storehouse |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202220385354.2U CN217699228U (en) | 2022-02-24 | 2022-02-24 | A micro-fluidic chip for unicellular automation is built storehouse |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN217699228U true CN217699228U (en) | 2022-11-01 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202220385354.2U Active CN217699228U (en) | 2022-02-24 | 2022-02-24 | A micro-fluidic chip for unicellular automation is built storehouse |
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| CN (1) | CN217699228U (en) |
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- 2022-02-24 CN CN202220385354.2U patent/CN217699228U/en active Active
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