CN214320197U - Single cell sorting micro-fluidic chip - Google Patents
Single cell sorting micro-fluidic chip Download PDFInfo
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- CN214320197U CN214320197U CN202022989247.4U CN202022989247U CN214320197U CN 214320197 U CN214320197 U CN 214320197U CN 202022989247 U CN202022989247 U CN 202022989247U CN 214320197 U CN214320197 U CN 214320197U
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Abstract
The utility model provides a micro-fluidic chip is selected separately to unicell, including the introduction port, go out the introduction port and connect gradually a plurality of capture units between introduction port and the introduction port, capture the unit and include cell flow channel, bypass the passageway, unicell capture passageway and unicell collection channel, wherein, cell flow channel's export, bypass the entry of passageway and unicell collection channel's entry link to each other in order to constitute tee bend structure, the entry of the cell flow channel of unit is captured to the back links to each other with the export of the bypass passageway of the preceding capture unit, the entry of the passageway is captured to the unicell and the pipe wall connection of the section of flowing into of bypass the passageway, the export of passageway is captured to the unicell and the pipe wall connection of the section of flowing out of bypass the passageway. The utility model discloses a single-celled interception can be realized to the size of control flow resistance and catch position, combines drive assembly, makes single-celled collection more convenient high-efficient. In addition, the micro-column array avoids blockage of subsequent channels by multi-cell clusters and impurities, and smoothness of single cell sorting is guaranteed.
Description
Technical Field
The utility model belongs to the technical field of micro-fluidic and cell are selected separately, a micro-fluidic chip is selected separately to unicellular is related to.
Background
Cells are basic units of life activities, and researches based on single cell level can reveal the development rule of life activities at a deeper level. Single cell sorting is the basis and key to single cell research. Currently, single cell sorting is mainly performed by a micro-needle aspiration method, microdissection, a limiting dilution method, a micropore array and a sorting method based on microfluidics. The current method has the problems of high operation difficulty, low efficiency, cell damage, multi-cell acquisition and the like, and is not beneficial to subsequent analysis. Therefore, a high-efficiency sorting method with simple operation, small cell damage and high single cell efficiency is needed in single cell research.
SUMMERY OF THE UTILITY MODEL
In view of the above shortcomings of the prior art, an object of the present invention is to provide a single cell sorting microfluidic chip for solving the problems of large operation difficulty, low efficiency, cell damage, multi-cell acquisition, etc. of the single cell sorting method in the prior art.
In order to achieve the above objects and other related objects, the present invention provides a single cell sorting microfluidic chip, including:
a sample inlet;
a sample outlet;
the multiple capturing units are sequentially connected between the sample inlet and the sample outlet, each capturing unit comprises a cell flow channel, a bypass channel, a single cell capturing channel and a single cell collecting channel, wherein the outlet of the cell flow channel, the inlet of the bypass channel and the inlet of the single cell collecting channel are connected to form a three-way structure, the inlet of the cell flow channel of the capturing unit is connected with the outlet of the bypass channel of the capturing unit, the inlet of the single cell capturing channel is connected with the pipe wall of the inflow section of the bypass channel, and the outlet of the single cell capturing channel is connected with the pipe wall of the outflow section of the bypass channel.
Optionally, the single-cell capture channel has an inlet size larger than an outlet size of the single-cell capture channel, and the outlet size of the single-cell capture channel is set smaller than a passage size of a single cell to be sorted.
Optionally, the single-cell capture channel has a smaller flow resistance than the bypass channel, and is configured to accommodate only a single cell to be sorted.
Optionally, the outlet of the single cell collecting channel is provided with a driving part for ejecting the captured single cell.
Optionally, the drive component comprises one of a thermal bubble printhead, a microvalve, and a solenoid valve.
Optionally, a transition channel is connected between the sample inlet and the first capturing unit, a micro-column array is arranged in the transition channel, and the micro-column array comprises a plurality of micro-columns arranged at intervals.
Optionally, the inlet size of the transition passage is larger than the outlet size of the transition passage.
Optionally, the cell flow channel is sized to allow passage of only a single cell to be sorted.
Optionally, the detour channel is bent back and forth at least once.
Optionally, the single cell collection channel is linear.
As above, the utility model discloses a micro-fluidic chip is selected separately to unicell accessible matches the flow resistance proportion of catching passageway and bypass passageway, makes the flow resistance of catching the passageway be less than the flow resistance of bypass passageway, and the size design of catching the position simultaneously realizes unicellular interception for only can holding single cell. Meanwhile, the cell collection is more convenient and efficient by combining with driving components such as a thermal bubble printing head, a PDMS micro valve, an electromagnetic valve and the like. In addition, the micro-column array avoids blockage of subsequent channels by multi-cell clusters and impurities, and smoothness of single cell sorting is guaranteed.
Drawings
Fig. 1 is a top view of the single-cell sorting microfluidic chip of the present invention.
FIG. 2 is a schematic diagram showing the capture of a single cell by the capture unit.
FIG. 3 is a schematic diagram showing the use of a thermal bubble printhead to accomplish the collection of single cells.
Description of the element reference numerals
1 sample inlet
2 sample outlet
3 Capture Unit
301 cell flow channel
302 detour channel
303 single cell trapping channel
304 single cell collection channel
4 flow passage plate
5 transition passage
6 micro-column array
7 drive component
8 single cell
Detailed Description
The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. The present invention can also be implemented or applied through other different specific embodiments, and various details in the present specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.
Please refer to fig. 1 to 3. It should be noted that the drawings provided in the present embodiment are only for illustrating the basic idea of the invention in a schematic manner, and only the components related to the invention are shown in the drawings rather than being drawn according to the number, shape and size of the components in actual implementation, and the form, quantity and proportion of the components in actual implementation may be changed at will, and the layout of the components may be more complicated.
Example one
In this embodiment, a single-cell sorting microfluidic chip is provided, please refer to fig. 1, which shows a top view of the single-cell sorting microfluidic chip, and includes a sample inlet 1, a sample outlet 2, and a plurality of capturing units 3, where the capturing units 3 are sequentially connected between the sample inlet 1 and the sample outlet 2.
Specifically, the sample inlet 1, the sample outlet 2 and the capturing unit 3 can be manufactured based on a flow channel plate 4, the flow channel plate can be of a multilayer structure, and the material of the flow channel plate includes but is not limited to silicon, acrylic and the like. Except for the sample inlet 1 and the sample outlet 2, the periphery of the channel of the capturing unit 3 can be in a closed state.
Specifically, the capturing unit 3 includes a cell flow channel 301, a bypass channel 302, a single cell capturing channel 303 and a single cell collecting channel 304, wherein an outlet of the cell flow channel 301, an inlet of the bypass channel 302 and an inlet of the single cell collecting channel 304 are connected to form a three-way structure, an inlet of the cell flow channel 301 of the latter capturing unit 3 is connected to an outlet of the bypass channel 302 of the former capturing unit 3, an inlet of the single cell capturing channel 303 is connected to a tube wall of an inflow section of the bypass channel 302, and an outlet of the single cell capturing channel 303 is connected to a tube wall of an outflow section of the bypass channel 302.
As an example, the passage size of the cell flow channel 301 is set to allow only a single cell to be sorted to pass through.
As an example, the flow resistance of the single-cell capturing channel 303 is set to be smaller than the flow resistance of the bypass channel 302, thereby ensuring that the flowing cells preferentially flow to the single-cell capturing channel 303 and realize cell interception.
Specifically, the flow resistance of the channel can be controlled by adjusting the channel size, the channel length, the pattern of the channels, and the like, for example, the longer the channel length, the larger the flow resistance.
By way of example, the bypass channel 302 is bent back and forth at least once to increase flow resistance. In this embodiment, the bypass channel 302 is U-shaped, the single-cell capture channel 303 is substantially linear, and the single-cell collection channel 304 is linear. In other embodiments, the shapes and sizes of the bypass channel 302, the single cell capturing channel 303 and the cell collecting channel 304 can be adjusted as required, and the protection scope of the present invention should not be limited herein.
As an example, the size of the inlet of the single-cell capture channel 303 is larger than the size of the outlet of the single-cell capture channel 303, and the size of the outlet of the single-cell capture channel 303 is set to be smaller than the passing size of a single cell to be sorted, so that a single cell cannot be discharged through the outlet of the single-cell capture channel 303 after entering the single-cell capture channel 303.
In this embodiment, the single-cell capturing channel 303 is set to only accommodate a single cell to be sorted, after the cell is intercepted, the flow resistance of the single-cell capturing channel 303 is increased, and the subsequent cell preferentially flows to the bypass channel 302, so as to ensure the single-cell intercepting accuracy in the single-cell capturing channel 303.
As an example, a transition channel 5 is connected between the sample inlet 1 and the first capture unit 3, in this embodiment, an inlet size of the transition channel 5 is larger than an outlet size of the transition channel 5, and a passing size of the transition channel 5 is gradually changed, which facilitates the gradual transition of the cell suspension flowing into the subsequent channel.
By way of example, a micro-column array 6 is arranged in the transition channel 5, and the micro-column array 6 includes a plurality of micro-columns arranged at intervals. The micro-column array 6 is used for intercepting impurities and multi-cell clusters and preventing the impurities and the multi-cell clusters from blocking subsequent channels. After the cell suspension has passed through the respective capturing unit 3, excess cells flow out of the outlet 2.
As an example, the outlet of the single cell collecting channel 304 is provided with a driving part 7 for ejecting the captured single cell. The driving member 7 includes, but is not limited to, one of a thermal bubble print head, a micro valve (e.g., a Polydimethylsiloxane (PDMS) micro valve), and a solenoid valve.
In this embodiment, the driving unit 7 is preferably a thermal bubble print head, which includes a nozzle connected to the outlet of the single-cell collecting channel 304 and a heating film integrated at the bottom of the channel, and the liquid above the heating film is vaporized by the instantaneous high temperature of the heating film, so that bubbles are generated to push the liquid to flow and be ejected from the nozzle, and then the subsequent liquid is replenished.
Specifically, the single-cell sorting microfluidic chip of the present embodiment separates single cells into two processes: (1) capturing single cells; (2) and (4) collecting single cells. Referring to fig. 2, a schematic diagram of the capturing of the single cell 8 by the capturing unit 3 is shown. Referring to fig. 3, a schematic diagram of the single cell 8 collection by the thermal bubble print head is shown, wherein the process of heating the membrane heating liquid to generate bubbles to push the liquid to flow carries the single cell 8 away from the interception position (the single cell capture channel 303), and ejects the single cell 8 from the nozzle into a designated container to achieve cell collection. The thermal bubble printing head has the advantages of fast response, strong driving force, convenient control, easy integration and miniaturization, and provides guarantee for convenient and efficient sorting of single cells.
The single-cell sorting microfluidic chip of the embodiment can enable the flow resistance of the capturing channel to be smaller than that of the bypassing channel by matching the flow resistance proportion of the capturing channel and the bypassing channel, and meanwhile, the size of the capturing position is designed to be capable of accommodating only a single cell, so that the single cell is intercepted. Meanwhile, the cell collection is more convenient and efficient by combining with driving components such as a thermal bubble printing head, a PDMS micro valve, an electromagnetic valve and the like. In addition, the micro-column array avoids blockage of subsequent channels by multi-cell clusters and impurities, and smoothness of single cell sorting is guaranteed.
Example two
This example used the microfluidic chip described in example one for single cell sorting.
Specifically, for a specific cell, the suspension of the cell is introduced into the microfluidic chip, and the micro-column array filters impurities and multi-cell clusters to ensure the flow of single cells in the subsequent channel. When the cells flow through the capturing unit, because the flow resistance of the single cell capturing channel is smaller than that of the bypass channel, the cells are preferentially intercepted in the single cell capturing channel, so that the flow resistance of the single cell capturing channel is increased, the subsequent cells can only flow to the next capturing unit through the bypass channel, and the capturing precision of the single cells is ensured. The design of the capture unit is closely related to the cell size. When the size of the cell changes, the width of the capturing channel is changed, so that the capturing position can only accommodate a single cell, and meanwhile, the length of the bypassing channel is changed, so that the flow resistance of the bypassing channel is greater than that of the capturing channel after the bypassing channel is changed, and the capturing of single cells with different sizes can be realized. After the cells are captured, the thermal bubble printing head below the cells is triggered to eject the cells from the jet orifice, so that the collection of the single cells is realized.
To sum up, the utility model discloses a micro-fluidic chip accessible is selected separately to unicell matches the flow resistance proportion of catching passageway and bypass channel, makes the flow resistance of catching the passageway be less than the flow resistance of bypass channel, and the size design of catching the position simultaneously realizes unicellular interception for only can holding single cell. Meanwhile, the cell collection is more convenient and efficient by combining with driving components such as a thermal bubble printing head, a PDMS micro valve, an electromagnetic valve and the like. In addition, the micro-column array avoids blockage of subsequent channels by multi-cell clusters and impurities, and smoothness of single cell sorting is guaranteed. Therefore, the utility model effectively overcomes various defects in the prior art and has high industrial utilization value.
The above embodiments are merely illustrative of the principles and effects of the present invention, and are not to be construed as limiting the invention. Modifications and variations can be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. A single-cell sorting microfluidic chip, comprising:
a sample inlet;
a sample outlet;
the multiple capturing units are sequentially connected between the sample inlet and the sample outlet, each capturing unit comprises a cell flow channel, a bypass channel, a single cell capturing channel and a single cell collecting channel, wherein the outlet of the cell flow channel, the inlet of the bypass channel and the inlet of the single cell collecting channel are connected to form a three-way structure, the inlet of the cell flow channel of the capturing unit is connected with the outlet of the bypass channel of the capturing unit, the inlet of the single cell capturing channel is connected with the pipe wall of the inflow section of the bypass channel, and the outlet of the single cell capturing channel is connected with the pipe wall of the outflow section of the bypass channel.
2. The single-cell sorting microfluidic chip of claim 1, wherein: the single cell capture channel has an inlet size larger than an outlet size of the single cell capture channel, and the outlet size of the single cell capture channel is set smaller than a pass size of a single cell to be sorted.
3. The single-cell sorting microfluidic chip of claim 1, wherein: the single-cell capture channel has a flow resistance less than that of the bypass channel, and is configured to accommodate only a single cell to be sorted.
4. The single-cell sorting microfluidic chip of claim 1, wherein: and a driving part for ejecting the captured single cells is arranged at the outlet of the single cell collecting channel.
5. The single-cell sorting microfluidic chip of claim 4, wherein: the driving part comprises one of a thermal bubble printing head, a micro valve and an electromagnetic valve.
6. The single-cell sorting microfluidic chip of claim 1, wherein: the sample inlet and the first are connected with a transition channel between the capture units, a micro-column array is arranged in the transition channel and comprises a plurality of micro-columns arranged at intervals.
7. The single-cell sorting microfluidic chip of claim 6, wherein: the inlet dimension of the transition passage is greater than the outlet dimension of the transition passage.
8. The single-cell sorting microfluidic chip of claim 1, wherein: the cell flow channel is sized to allow passage of only a single cell to be sorted.
9. The single-cell sorting microfluidic chip of claim 1, wherein: the detour channel is bent back and forth at least once.
10. The single-cell sorting microfluidic chip of claim 1, wherein: the single cell collecting channel is linear.
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| CN202022989247.4U CN214320197U (en) | 2020-12-11 | 2020-12-11 | Single cell sorting micro-fluidic chip |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023184647A1 (en) * | 2022-03-31 | 2023-10-05 | 上海傲睿科技有限公司 | Microfluidic chip and methods for culturing single cell and screening and exporting cell population |
| WO2023201942A1 (en) * | 2022-04-22 | 2023-10-26 | 上海傲睿科技有限公司 | Single-cell sorting microfluidic chip and single-cell sorting method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023184647A1 (en) * | 2022-03-31 | 2023-10-05 | 上海傲睿科技有限公司 | Microfluidic chip and methods for culturing single cell and screening and exporting cell population |
| WO2023201942A1 (en) * | 2022-04-22 | 2023-10-26 | 上海傲睿科技有限公司 | Single-cell sorting microfluidic chip and single-cell sorting method |
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