A single-cell sorting microfluidic chip

Abstract

The present invention provides a single-cell sorting microfluidic chip, which includes a sample inlet, a sample outlet, and a plurality of capture units sequentially connected between the sample inlet and the sample outlet. The capture unit includes a cell flow channel, a bypass channel, a single-cell capture channel, and a single-cell collection channel. Among them, the outlet of the cell flow channel, the inlet of the bypass channel, and the inlet of the single-cell collection channel are connected to form a tee structure. The inlet of the cell flow channel of the latter capture unit is connected to the outlet of the bypass channel of the previous capture unit. The inlet of the single-cell capture channel is connected to the tube wall of the inflow section of the bypass channel, and the outlet of the single-cell capture channel is connected to the tube wall of the outflow section of the bypass channel. By controlling the flow resistance and the size of the capture position, the present invention can achieve the interception of single cells. At the same time, combined with the driving component, the single-cell collection is made more convenient and efficient. In addition, the microcolumn array avoids the blockage of the subsequent channels by multi-cell clusters and impurities, ensuring the smoothness of single-cell sorting.

Description

Technical Field

[0001] The present invention belongs to the technical fields of microfluidics and cell sorting, and relates to a microfluidic chip for single-cell sorting. Background Art

[0002] Cells are the basic units of life activities. Research at the single-cell level can reveal the development laws of life activities at a deeper level. Single-cell sorting is the basis and key of single-cell research. Currently, single-cell sorting mainly includes micro-needle aspiration, microdissection, limited dilution, micro-well arrays, and microfluidics-based sorting methods. The current methods face problems such as high operation difficulty, low efficiency, cell damage, and multi-cell acquisition, which are not conducive to subsequent analysis. Therefore, there is an urgent need for an efficient sorting method with simple operation, small cell damage, and high single-cell rate in single-cell research. Summary of the Invention

[0003] In view of the above-mentioned disadvantages of the prior art, the purpose of the present invention is to provide a microfluidic chip for single-cell sorting, which is used to solve the problems of high operation difficulty, low efficiency, cell damage, and multi-cell acquisition in the single-cell sorting method of the prior art.

[0004] To achieve the above purpose and other related purposes, the present invention provides a microfluidic chip for single-cell sorting, including:

[0005] An inlet;

[0006] An outlet;

[0007] A plurality of capture units, sequentially connected between the inlet and the outlet. The capture unit includes a cell flow channel, a bypass channel, a single-cell capture channel, and a single-cell collection channel. Among them, the outlet of the cell flow channel, the inlet of the bypass channel, and the inlet of the single-cell collection channel are connected to form a tee structure. The inlet of the cell flow channel of the latter capture unit is connected to the outlet of the bypass channel of the previous capture unit. The inlet of the single-cell capture channel is connected to the tube wall of the inflow section of the bypass channel, and the outlet of the single-cell capture channel is connected to the tube wall of the outflow section of the bypass channel.

[0008] Optionally, the inlet size of the single-cell capture channel is larger than the outlet size of the single-cell capture channel, and the outlet size of the single-cell capture channel is set to be smaller than the passing size of a single cell to be sorted.

[0009] Optionally, the flow resistance of the single-cell capture channel is less than the flow resistance of the bypass channel, and the single-cell capture channel is set to only accommodate a single cell to be sorted.

[0010] Optionally, a driving component for ejecting the captured single cell is provided at the outlet of the single-cell collection channel.

[0011] Optionally, the driving component includes one of a thermal bubble print head, a microvalve, and a solenoid valve.

[0012] Optionally, a transition channel is connected between the sample inlet and the first capture unit, and a microcolumn array is provided in the transition channel. The microcolumn array includes a plurality of microcolumns arranged at intervals.

[0013] Optionally, the inlet size of the transition channel is larger than the outlet size of the transition channel.

[0014] Optionally, the passing size of the cell flow channel is set to allow only a single cell to be sorted to pass through.

[0015] Optionally, the detour channel bends back and forth at least once.

[0016] Optionally, the single-cell collection channel is linear.

[0017] As described above, the single-cell sorting microfluidic chip of the present invention can match the flow resistance ratio of the capture channel and the detour channel, so that the flow resistance of the capture channel is less than that of the detour channel. At the same time, the size of the capture position is designed to accommodate only a single cell, realizing the interception of single cells. Meanwhile, combined with driving components such as a thermal bubble print head, a PDMS microvalve, a solenoid valve, etc., cell collection is made more convenient and efficient. In addition, the microcolumn array avoids the blockage of subsequent channels by multi-cell clusters and impurities, ensuring the smoothness of single-cell sorting. Description of the Drawings

[0018] Figure 1 Shown is a top view of the single-cell sorting microfluidic chip of the present invention.

[0019] Figure 2 Shown is a schematic diagram of first completing the capture of a single cell by the capture unit.

[0020] Figure 3 Shown is a schematic diagram of completing the collection of a single cell by a thermal bubble print head.

[0021] Description of Reference Numerals

[0022] 1 Sample inlet

[0023] 2 Sample outlet

[0024] 3 Capture unit

[0025] 301 Cell flow channel

[0026] 302 Detour channel

[0027] 303 Single-cell capture channel

[0028] 304 Single-cell collection channel

[0029] 4 Flow channel plate

[0030] 5 Transition channel

[0031] 6 Microcolumn array

[0032] 7 Driving component

[0033] 8 Single cell Detailed implementation manners

[0034] The following uses specific specific examples to illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0035] Please refer to Figures 1 to 3 . It should be noted that the illustrations provided in this embodiment only illustrate the basic concept of the present invention in a schematic manner. Therefore, only the components related to the present invention are shown in the drawings, rather than being drawn according to the number, shape, and size of the components in actual implementation. The type, quantity, and ratio of each component in its actual implementation can be arbitrarily changed, and the component layout type may also be more complex.

[0036] Example 1

[0037] In this embodiment, a single-cell sorting microfluidic chip is provided. Please refer to Figure 1 , which shows a top view of the single-cell sorting microfluidic chip, including a sample inlet 1, a sample outlet 2, and a plurality of capture units 3. The plurality of capture units 3 are sequentially connected between the sample inlet 1 and the sample outlet 2.

[0038] Specifically, the sample inlet 1, the sample outlet 2, and the capture unit 3 can be fabricated based on a flow channel plate 4. The flow channel plate can be a multi-layer structure, and its materials include but are not limited to silicon, acrylic, etc. Except for the sample inlet 1 and the sample outlet 2, the channels around the capture unit 3 can be in a closed state.

[0039] Specifically, the capture unit 3 includes a cell flow channel 301, a bypass channel 302, a single-cell capture channel 303, and a single-cell collection channel 304. Among them, the outlet of the cell flow channel 301, the inlet of the bypass channel 302, and the inlet of the single-cell collection channel 304 are connected to form a tee structure. The inlet of the cell flow channel 301 of the latter capture unit 3 is connected to the outlet of the bypass channel 302 of the previous capture unit 3. The inlet of the single-cell capture channel 303 is connected to the tube wall of the inflow section of the bypass channel 302, and the outlet of the single-cell capture channel 303 is connected to the tube wall of the outflow section of the bypass channel 302.

[0040] 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.

[0041] As an example, the flow resistance of the single-cell capture channel 303 is set to be less than that of the bypass channel 302, so as to ensure that the flowing cells preferentially flow to the single-cell capture channel 303 and achieve cell interception.

[0042] Specifically, the flow resistance of the channel can be controlled by adjusting the channel size, channel length, and channel pattern, etc. For example, the longer the channel length, the greater the flow resistance.

[0043] As an example, the bypass channel 302 bends back and forth at least once to increase the flow resistance. In this embodiment, the bypass channel 302 is U-shaped, the single-cell capture channel 303 is generally linear, and the single-cell collection channel 304 is linear. In other embodiments, the patterns and sizes of the bypass channel 302, the single-cell capture channel 303, and the cell collection channel 304 can be adjusted as needed, and the protection scope of the present invention should not be unduly limited here.

[0044] As an example, the inlet size of the single-cell capture channel 303 is larger than the outlet size of the single-cell capture channel 303, and the outlet size of the single-cell capture channel 303 is set to be smaller than the passage size of a single cell to be sorted, so that the single cell cannot be discharged through the outlet of the single-cell capture channel 303 after entering the single-cell capture channel 303.

[0045] In this embodiment, the single-cell capture channel 303 is set to be able to accommodate only a single cell to be sorted. After the cell is intercepted, the flow resistance of the single-cell capture channel 303 increases, and the subsequent cells preferentially flow to the bypass channel 302 to ensure the single-cell interception accuracy in the single-cell capture channel 303.

[0046] As an example, a transition channel 5 is connected between the sample inlet 1 and the first capture unit 3. In this embodiment, the inlet size of the transition channel 5 is larger than the outlet size of the transition channel 5, and the passing size of the transition channel 5 is gradually changing, which is conducive to the gradual transition of the cell suspension into the subsequent channels.

[0047] As an example, a microcolumn array 6 is provided in the transition channel 5. The microcolumn array 6 includes a plurality of microcolumns arranged at intervals. The microcolumn array 6 is used to intercept impurities and multicellular clusters to prevent them from blocking the subsequent channels. After the cell suspension passes through each capture unit 3, the excess cells flow out from the sample outlet 2.

[0048] As an example, a driving component 7 for ejecting the captured single cells is provided at the outlet of the single cell collection channel 304. The driving component 7 includes, but is not limited to, one of a thermal bubble print head, a microvalve (such as a polydimethylsiloxane PDMS microvalve), and a solenoid valve.

[0049] In this embodiment, the driving component 7 preferably adopts a thermal bubble print head, which includes a nozzle connected to the outlet of the single cell collection channel 304 and a heating film integrated at the bottom of the channel. The heating film uses the instant high temperature to vaporize the liquid above, thereby generating bubbles to push the liquid to flow and eject from the nozzle, and then the subsequent liquid is replenished.

[0050] Specifically, the single cell sorting microfluidic chip of this embodiment sorts single cells into two processes: (1) single cell capture; (2) single cell collection. Please refer to Figure 2 which shows a schematic diagram of first completing the capture of the single cell 8 by the capture unit 3. Please refer to Figure 3 which shows a schematic diagram of using a thermal bubble print head to complete the collection of the single cell 8. Among them, in the process that the heating film heats the liquid to generate bubbles to push the liquid to flow, the single cell 8 is taken away from the interception position (the single cell capture channel 303) and ejected from the nozzle into the specified container to achieve cell collection. The thermal bubble print head has the advantages of fast response, strong driving force, easy control, easy integration and miniaturization, providing guarantee for the convenient and efficient sorting of single cells.

[0051] The single cell sorting microfluidic chip of this embodiment can match the flow resistance ratio of the capture channel and the bypass channel, make the flow resistance of the capture channel smaller than that of the bypass channel, and at the same time, the size of the capture position is designed to only accommodate a single cell to achieve the interception of single cells. At the same time, combined with driving components, such as thermal bubble print heads, PDMS microvalves, solenoid valves, etc., the cell collection is made more convenient and efficient. In addition, the microcolumn array avoids the blockage of the subsequent channels by multicellular clusters and impurities, ensuring the smoothness of single cell sorting.

[0052] Embodiment Two

[0053] In this embodiment, the microfluidic chip described in Embodiment 1 is used for single-cell sorting.

[0054] Specifically, for a certain specific type of cell, the suspension of this cell is introduced into the microfluidic chip. The micro-column array filters out impurities and multi-cell clusters to ensure the flow of single cells in the subsequent channels. When the cell flows through the capture unit, since the flow resistance of the single-cell capture channel is less than that of the bypass channel, the cell is preferentially intercepted in the single-cell capture channel, resulting in an increase in the flow resistance of the single-cell capture channel. Subsequently, the cells can only flow through the bypass channel to the next capture unit, ensuring the capture accuracy of single cells. The design of the capture unit is closely related to the cell size. When the cell size changes, the width of the capture channel is changed so that the capture position can only accommodate a single cell. At the same time, the length of the bypass channel is changed so that its flow resistance is greater than that of the changed capture channel, thus enabling the capture of single cells of different sizes. After the cell is captured, the thermal bubble printhead below is triggered to eject the cell from the nozzle hole to achieve the collection of single cells.

[0055] In summary, the single-cell sorting microfluidic chip of the present invention can achieve the interception of single cells by matching the flow resistance ratio of the capture channel and the bypass channel, making the flow resistance of the capture channel less than that of the bypass channel, and at the same time, the size of the capture position is designed to only accommodate a single cell. Meanwhile, combined with driving components such as thermal bubble printheads, PDMS microvalves, solenoid valves, etc., the cell collection is made more convenient and efficient. In addition, the micro-column array avoids the blockage of the subsequent channels by multi-cell clusters and impurities, ensuring the smoothness of single-cell sorting. Therefore, the present invention effectively overcomes various drawbacks in the prior art and has high industrial utilization value.

[0056] The above embodiments are only illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed by the present invention should still be covered by the claims of the present invention.

Claims

1. A single-cell sorting microfluidic chip, characterized in that Comprising: Sampling inlet; Sampling outlet; A plurality of capture units, sequentially connected between the sampling inlet and the sampling outlet. The capture unit includes a cell flow channel, a detour channel, a single-cell capture channel and a single-cell collection channel. Among them, the outlet of the cell flow channel, the inlet of the detour channel and the inlet of the single-cell collection channel are connected to form a tee structure. The inlet of the cell flow channel of the latter capture unit is connected to the outlet of the detour channel of the previous capture unit. The inlet of the single-cell capture channel is connected to the tube wall of the inflow section of the detour channel, and the outlet of the single-cell capture channel is connected to the tube wall of the outflow section of the detour channel; Among them, a driving component for ejecting the captured single cell is provided at the outlet of the single-cell collection channel. The driving component adopts a thermal bubble print head. The thermal bubble print head includes a nozzle connected to the outlet of the single-cell collection channel and a heating film integrated at the bottom of the channel. The heating film heats the liquid to generate bubbles to push the liquid to flow, so as to take the single cell away from the single-cell capture channel and eject the single cell from the nozzle into a specified container to realize cell collection.

2. The single-cell sorting microfluidic chip according to claim 1, wherein: The inlet size of the single-cell capture channel is larger than the outlet size of the single-cell capture channel, and the outlet size of the single-cell capture channel is set to be smaller than the passing size of a single cell to be sorted.

3. The single-cell sorting microfluidic chip according to claim 1, wherein: The flow resistance of the single-cell capture channel is smaller than that of the detour channel, and the single-cell capture channel is set to only accommodate a single cell to be sorted.

4. The single-cell sorting microfluidic chip according to claim 1, characterized in that: A transition channel is connected between the sampling inlet and the first capture unit. A micro-column array is provided in the transition channel. The micro-column array includes a plurality of micro-columns arranged at intervals.

5. The single-cell sorting microfluidic chip according to claim 4, wherein: The inlet size of the transition channel is larger than the outlet size of the transition channel.

6. The single-cell sorting microfluidic chip according to claim 1, wherein: The passing size of the cell flow channel is set to only allow a single cell to be sorted to pass through.

7. The single-cell sorting microfluidic chip according to claim 1, characterized in that: The detour channel bends back and forth at least once.

8. The single-cell sorting microfluidic chip according to claim 1, wherein: The single-cell collection channel is linear.

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